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LIBRARY
Michigan State
University

 

 

This is to certify that the
thesis entitled

Taxonomic Implications of Distribution,
Breeding Systems, Vestiture Type, and Pollen

Morphology in Selected Mexican Hieracium Species
presented by

Jacqueline Anne Soule

has been accepted towards fulfillment
of the requirements for

Masters of Sciencedegree in Botany

 

 

Major professor

Date A? J”?! 41%?

0-7639 MS U i: an Affirmative Action/Equal Opportunity Institution

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PLACE IN RETURN BOX to remove this checkout from your record.
TO AVOID FINES return on or betore date due.

DATE DUE DATE DUE DATE DUE
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MSU I: An Affirmative Action/Equal Opportunity Institution
cmmflf

 

TAXONOMIC IMPLICATIONS OF DISTRIBUTION,
BREEDING SYSTEMS, VESTITURE TYPE, AND POLLEN MORPHOLOGY

IN SELECTED MEXICAN HIERACIUM SPECIES (COMPOSITAE)
By

Jacqueline Anne Soule

A THESIS

Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of

MASTER OF SCIENCE

Department of Botany and Plant Pathology

1988

ABSTRACT

TAXONOMIC IMPLICATIONS OF DISTRIBUTION, BREEDING SYSTEMS, VESTITURE TYPE,
AND POLLEN MORPHOLOGY IN SELECTED MEXICAN HIERACIUM SPECIES (COMPOSITAE)

By

Jacqueline Anne Soule

The taxonomic relationships of the polytypic Mexican Hawkweeds,
Hieracium abscissum Less., fl. dysonymum Blake,.§. mexicanum Less., fl.
pringlei A. Gray, 3. schultzii Fries, and two potentially new taxa,

fl, 'potosi' and fl, 'chipingue' were addressed in this paper. Extensive
field work discovered no clearly intermediate or hybrid individuals,
indicating that the polytypy observed in herbarium specimens is more
likely due to responses to different sites than to hybridization.
Breeding system studies which included pollinator exclusion in-situ,
emasculation, and artificial selfing indicate that these Hieracium
species are outcrossers. A vestiture survey based on scanning electron
microscopy (SEM) and analysis by Spearman's Rank correlation and Average
Linkage Cluster Analysis resulted in the taxa sorting into two groups,
the Hieracium abscissum-schultzii group to which E, 'chipingue' belongs,
and the Hieracium dysonymum-mexicanum group to which E, 'potosi'
belongs. SEM studies of pollen morphology determined two groups which
correspond to the plants' growth form. The many headed species -—
Hieracium abscissum, fl, schultzii, and fl. 'chipingue,’ have curved
spines; the few headed species -- fl, dysonymum, fl, mexicanum,

_fl. pringlei and fl. 'potosi,' have straight spines. The number of
annular and lacunar pollen spines provides evidence for not recognizing
Hieracium 'chipinque' and fl, 'potosi' as separate taxa since these
pollen characters agree with other characters, particularly vestiture,

of E. abscissum and fl, dysonymum respectively.

DEDICATION

To the people whose love, encouragement, and belief in me
made this dream a reality; my grandfather and grandmother,

my father, my brother, and my mentor.

ii

ACKNOWLEDGMENTS

I wish to express my gratitude to all those who provided guidance,
help, and support during this study. My thanks especially to Dr. John H.
Beaman, my major professor, for his efforts, and to my committee members,
Dr. Karen Klomparens and Dr. Stephen Stephenson for their invaluable
input. I am also indebted to Dr. Peter Murphy who provided me with space
for growing plants. Technical assistance was freely given by the members
of Dr. Chris Stevens', and Dr. Barbara Sears' labs and by the staff at the
Center for Electron Optics, for which I thank them all.

Of course, none of this would have been possible without the people
who helped me find my way, and all the people at MSU who became my friends
and gave me the support, encouragement, spelling corrections, and suitable
libations as the occasion warranted. I thank them all and list them below
in no particular order: Jeremy Soule, Cynthia Baker, Thomas Harland III,
Graham Smart, Sara Kaplan, Koibeatu, Gerald 'Stinger' Guala, Tom Trana,
Shirley Owens, Chris Pedersen, Tracy Wacker, Carolyn Pazur, Phil Jensen,
Peggy Hogan, Tom Isakiet, Barry Stein, Kate Everts, and the attendees of
the Plant Diversity Discussion Group and the Alex B. Filanow Memorial
Symposium.

Collecting expeditions to Mexico were made possible in part by funds

from the William T. Gillis Memorial Fund and an award from the Michigan

State University Chapter of the Society of The Sigma Xi.

iii

TABLE

LIST OF TABLES. . . . . . . . . .

LIST OF FIGURES

GENERAL INTRODUCTION. . . . . . .

OF CONTENTS

DISTRIBUTION, ECOLOGY, AND GEOLOGY.

Introduction . . . . .
MethOds. O O O O O O 0
Results and Discussion
Conclusions. . . . . .
BREEDING SYSTEM STUDIES . .
Introduction . . . . .
Methods. . . . . . . .
Results and Discussion
Conclusions. . . . . .
VESTITURE STUDIES . . . . .
Introduction . . . . .
Methods. . . . . . . .

Results and Discussion
Conclusions. . . . . .

POLLEN STUDIES. . . . . . .

Introduction . . . . .
Methods. . . . . . . .
Results and Discussion
Conclusions. . . . . .

BIBLImRAPHY. O O O O O O O
APPENDICES O O O O O O O O 0
Appendix I. Specimens

O O O I
O O O O
O O O O
O O O O
O O O O
O O O O
O O O O
O O O O
O O O O
O C O 0
used in

distribution maps
Appendix II. Raw data utilized

the vestiture . . . .

the compilation of

in statistical analysis

of

iv

35
35
36
38
65
66
66
68
69
78
79

87

87

97

LIST OF TABLES

Table Page

1. Hieracium taxa found growing together in the field and
still differentiatable as to species . . . . . . . . . . . . 13

2. Results of transects used to determine type of dispersion

Within a habitat O I O O O O C I O C O O C O O O O O O O O O 14
3. Chromosome counts of Mexican species of Hieracium. . . . . . 20
4. Results of pollinator exclusion study. . . . . . . . . . . . 27

5. Results of anther excision (emasculation) in selected
species of Mexican Hieracium . . . . . . . . . . . . . . . . 28

6. Results of artificial self-pollination in selected

species of Mexican Hieracium . . . . . . . . . . . . . . . . 29
7. Control group for breeding system studies in Mexican

HieraCium O O O O O O O O O O O O O O O O O O O O O O O O O 30

8. Counts of the number of seeds (fruits) per head in
field-collected heads of Mexican Hieracium . . . . . . . . . 31

9. Spearman's Rank Corelation Coefficients for the vestiture
data of all the taxa in the study. . . . . . . . . . . . . . 56

10. Spearman's Rank Correlation Cofficients for the vestiture
data of Hieracium abscissum, H. schultzii, and

fl.'ChiEinnge'.....oo..o......o...... 57

11. Spearman's Rank Correlation Cofficients for the vestiture
data of Hieracium dysonymum, H. pringlei, and
fl. 'EOtOSi' e e e e e e e e e o e o e e e o e e o o e e e o e 57

 

 

12. Spearman's Rank Correlation Cofficients for the vestiture
data of Hieracium dysonymum, fl.mexicanum, and
fl. 'EOtOSi' O o e e e o e e e e o e o O o o o e e o o o e e o 57

 

13. Average Linkage Cluster Analysis of vestiture data of all
taxa of Hieracium in the study . . . . . . . . . . . . . . . 58

V

LIST OF TABLES
(continued)

Table Page

14. Average Linkage Cluster Analysis of vestiture data for
Hieracium abscissum, H. schultzii, and H. 'chipingue'. . . . 60

 

15. Average Linkage Cluster Analysis of vestiture data for
Hieracium dySonymum, fl, pringlei, and fl. 'potosi'. . . . . . 61

16. Average Linkage Cluster Analysis of vestiture data for
Hieracium dysonymum, fl. mexicanum, and fl, 'potosi' . . . . . 62

17. Evaluations of species of Mexican Hieracium pollen
grains based on scanning electron micrographs. . . . . . . . 74

vi

10.

ll.

12.

13.

14.

LIST OF

Distribution map of Hieracium

1L 'chipingue'. . . . . . . .

Distribution map of Hieracium

FIGURES

abscissum and

dysonymum and

 

fl. 'EOtOSi ' O O O O I O O O 0

Distribution map of Hieracium

Distribution map of Hieracium

mexicanum . . . . .

pringlei. . . . . .

 

Distribution map of Hieracium

schultzii . . . . .

 

Scanning electron micrographs
Hieracium abscissum . . . . .

 

Scanning electron micrographs
Hieracium dysonymum . . . . .

Scanning electron micrographs
Hieracium mexicanum . . . . .

 

Scanning electron micrographs
Hieracium pringlei. . . . . .

 

Scanning electron micrographs
Hieracium schultzii . . . . .

 

Scanning electron micrographs
Hieracium 'chipinque' . . . .

Scanning electron micrographs
Hieracium 'potosi'. . . . . .

of the vestiture of

of the vestiture of

of the vestiture of

of the vestiture of

Dendrogram based on the Cluster Analysis of vestiture
data for all OTU's of Mexican Hieracium in this study

Dendrogram based on the Cluster Analysis of vestiture

data for Hieracium abscissum,

 

H- M and

fl. 'ChiEiflgue’ e c e e e e e e e o e e e e e e o e e 0

vii

Page

10
ll

12

41

43

45

47

49

51

53

59

6O

LIST OF FIGURES
(continued)

Figure Page

15. Dendrogram based on the Cluster Analysis of vestiture
data for Hieracium dysonymum, H. pringlei, and
fl. 'EOtOSi' O O O I C O O O O O O I O O C C O O C O O O O O 61

 

l6. Dendrogram based on the Cluster Analysis of vestiture
data for Hieracium dysonymum, H. mexicanum, and

fl. ' EOtOSi ' O O O O O O O O O O O O O O O O I O O O O O O O 62

 

17. Scanning electron micrographs of the pollen of
selected species of Mexican Hieracium . . . . . . . . . . . 71

18. Scanning electron micrographs of germinated pollen

from selected species of Mexican Hieracium. . . . . . . . . 73
19. Average size of Hieracium pollen grains by taxa . . . . . . 76

viii

INTRODUCTION

The genus Hieracium (Compositae, tribe Lactuceae) includes
herbaceous perennial species found in boreal and temperate regions of
Europe, Asia, North and South America. The generic epithet comes from

"...the ancients as

the Greek higggx: a hawk, and comes to us from
recorded by Pliny and others, supposing that the hawks used the plant to
strengthen their eyesight" (Fernald 1950). The common name is Hawkweed.
Over the years Hieracium has gained notoriety for being one of the
most taxonomically difficult of all genera. In the European species,
polyploidy, apomixis and hybridization have been shown to be the major
factors contributing to the biological complexity of the genus and have
led to a proliferation of recognized species. As Gleason (1952) states,

"...nearly 10,000 described species...of which perhaps only

there are
200-300 should be maintained".

The last published revision on the genus in Mexico is that of
Robinson and Greenman (1904). In a study of the genus in Mexico and
Central America, Beaman (unpublished) has determined that there are
approximately 18 species in that area. Six species need further work
before a definitive statement about the taxonomy of the genus in Mexico
and Central America can be made.

This study was limited to the problematical species found in

eastern and central Mexico that have shown the greatest polytypy,

Hieracium abscissum Less., H. dysonymum Blake, H. mexicanum Less.,

 

Hp pringlei A. Gray, and H. schultzii Fries. These species are all
widespread, and putative hybrids involving all of them were suspected on
the basis of herbarium specimens. Two areas in particular had plants
that were hard to definitively place to species. These areas
represented potential northward extensions of range for known species or
else sites of new species. This study also addressed the issue of
affiliation for the plants from Chipinque Mesa, above Monterrey, Nuevo
Lébn, and Cerro Potosi: Nuevo Lebn. These plants are referred to as

Hieracium 'chipinque' and H, 'potosi.’

 

Little experimental work has been done on the Mexican species.
Chromosome counts have been reported by Beaman and other workers, and
the numbers are only diploid with 2.= 9. Thus, the complex variation
may involve hybridization or inbreeding rather than polyploidy.
Conversely, it might be due to ecotypical variation or adaptive
radiation.

Four approaches to the problem were utilized: evaluation of
geographic distribution patterns, breeding system studies, examination
of the vestiture, and pollen analysis. Field work was conducted to
search for range extensions, determine dispersion within a habitat, and
evaluate localities where two or more Hieracium species grow together.
The breeding system studies involved field work and hybridization of
field-collected plants. The vestiture was examined by scanning electron
microscopy (SEM) and the data statistically analyzed by two techniques.

The pollen analysis utilized SEM on mostly field-collected pollen.

DISTRIBUTION, ECDLOGY, AND GEOLOGY

Introduction

 

Hieracium is primarily a temperate region genus that generally
does not tolerate aridity, and thus in Mexico is found mostly in the
highlands. These highlands range between 1,300 to 4,000 meters
elevation, and are generally covered with pine-oak or pine forests
(Miranda and Sharp 1950, Rzedowski 1978). Collections of the taxa
addressed in this work show them to be found mostly in the northern
Sierra Madre Oriental, the trans-Mexican volcanic range, and the Sierra
Madre del Sur in Oaxaca and Chiapas. When label data were accumulated
for preliminary location maps, the genus appeared disjunct, centering
mostly around cities with universities or in highly accessible scenic
areas. The separate collections of R. McVaugh, P. S. Martin, and J. H.
Beaman from remote areas indicated that the species might have
continuous ranges rather than disjunct ones.

Some species of Hieracium, however, are known to be disjunct,
notably H, mexicanum which is only found at higher elevations,

frequently above timberline, in the trans-Mexican volcanic range and

south. Hieracium abscissum, H. dysonymum, and fl. schultzii are found in

 

slightly lower elevation moist pine-oak woods, although H, dysonymum has

also been collected above timberline. Hieracium pringlei seems the most

tolerant of aridity and is found at the lowest elevations, often in
fairly xeric oak forests.

The two potentially new taxa, Hieracium 'potosi' and H. 'chipingue'
are known only from single local areas. The parent material for the
soil in both these areas is limestone (Beaman and Andresen 1966; B. L.
Turner, personal communication). In most other areas in Mexico where
Hieracium is found the soil is of volcanic origin (Beaman 1962, 1965;

Jardel 1986; Flores 1985). It was suspected that Hieracium 'potosi' and

 

.H. 'chipingue' were endemic to limestone, edaphic endemism being common
in many species (McVaugh 1943, 1952; Kruckeberg 1951, 1969; Braun 1955).

Hieracium pringlei has been collected on limestone outcrops as well as

 

from volcanic areas. Further collections of Hieracium lpotosi' and_H.

 

'chipingue' were needed to determine if they were limestone endemic
species, or whether, like H, pringlei, they represent known species
which occur on various soil types.

The work of Clausen, Keck, and Hiesey (l941)‘demonstrates how
greatly a taxon can vary based on the site to which it has adapted.
Their demonstration of variations between populations based on
topography indicates how careful systematists must be when recognizing
new taxa. Ecological studies have been demonstrated to be useful in
plant systematics, and were discussed at length by Kruckeberg (1969A).
He indicates that extensive collecting with great attention to detail,
such as soils and cover, can provide valuable ecological information on
a group and eliminate problems of topographic variants. Field work was
undertaken in part to determine the ranges of the species and to fill in

the gaps in range maps, or to demonstrate the disjunction of the ranges.

An additional study was done to determine the type of dispersion within

habitat and to see if it would shed light on the biology of Hieracium.

Methods

Specimens of Mexican Hieracium species were examined at the
following herbaria: ARIZ, ASUA, MEXU, MICH, MSC, M0, TEX, and XAL
(abbreviations after Holmgren et al. 1981). Also viewed were a large
number of specimens from WIS that were on loan to Dr. J. Beaman, as well
as his collections which have not yet been distributed. All label data
were recorded from these specimens, in part to establish what sites had
been collected, to determine where the species showed the greatest
polytypy, and for the formulation of location and distribution maps
(Figures 1 through 5). Location maps were compiled based on the
information on herbarium labels. These maps were taken into the field
and gaps between locations were extensively searched for the presence of
any of the species. Sites where two or more of the polytypic species
occurred were especially searched for. Distribution maps were compiled
once all field work was completed and include the author's collections.
Voucher specimens are in the Beal-Darlington Herbarium, Michigan State
University (MSC).

Two field trips to Mexico were made. In 1986 approximately 50
sites were examined in likely habitats in the trans-Mexican volcanic
belt. Prior collection localities were searched and attempts made to
find new localities. Eighteen sites were found to contain the genus and

were thus considered "collection sites." Collection sites were

evaluated for: dominant vegetation, possible plant associations, cover,
exposure, and elevation. Herbarium specimens, seeds, and where
possible, live plants were collected. Photographs of habitat and
individuals were taken at each site when weather permitted.

In 1987 roughly 400 sites were searched in likely-looking habitats
throughout the Sierra Madre Oriental and the trans-Mexican volcanic
belt. Fifty-three sites proved to contain the genus and are thus
designated "collection sites." Collection sites in 1987 were evaluated
for: slope (facing and degree), soil type, dominant vegetation, possible
plant associations, cover, exposure, and elevation. Herbarium
specimens, and where possible, live plants, pollen samples, and seeds
were collected. Photographs of habitat and individuals were taken at
each site when weather permitted.

Dispersion patterns within a locality were evaluated by line
transect at five sites. The line transect was selected at a site once
the first Hieracium individuals were found. This,was designated the
starting point of a 100 meter transect. The transect was laid out by
uncoiling a 100 m tape measure. Due to the nature of the habitats in
which Hieracium is found (steep mountain slopes, or often along the
edges of cliffs or roadside cuts), a totally random direction for the
transect could not be made. Instead, the worker faced up the slope and
made the decision to run the transect to the left or right based on a
coin toss; heads to the right, tails to the left. At the zero point and
at each ten meter interval, a one meter square quadrat was established
centered on the tape used to establish the transect. Within each of the

10 quadrats the number of individuals present and their age class was

recorded. The three age classes were: 1) seedling (four or less leaves
and small stature); 2) non-reproducing (more than four leaves and/or the

size of a reproducing plant but lacking flower stalk); 3) reproducing.

Results and Discussion

The summer field work was successful in filling some of the gaps in
the known distributions of Hieracium species in Mexico. The
distribution maps (Figures 1 through 5) are compiled in part from the
collections made by Soule et al. in 1986 and by Soule and Bodeau in
1987. The specimens of other collectors, particularly Beaman from 1957
to 1978, were also used. Specimens used for the maps are listed in
Appendix I. These maps indicate that the northern Sierra Madre Oriental

populations of Hieracium abscissum, and to a lesser degree fl, dysonymum

 

and H. schultzii, are disjunct from those in the trans-Mexican volcanic

range. Hieracium pringlei has an almost continuous distribution;

 

/
indeed, further collections in the states of San Luis Potosi and Durango

may show this to be so.

Soils

Soil evaluation at each collection site indicates that Hieracium
species are found on diverse soil types: H. abscissum, H. dysonymum, and
H, schultzii were found growing in clay, rhyolitic, and volcanic ash

soils; H. pringlei was found in limestone, gypsum, rhyolitic, clay, and

 

 

 

 

 

 

 

 

 

 

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Table 2. Results of transects used to determine type of dispersion within
a habitat.

Soule Number of Individuals Present per Quadrat Total
Coll. #/ Age
# Age Class Q1 Q2 Q3 Q4 Q5 Q6 Q7 Q8 Q9 Q10 Class
1501 Reproducing l 2 0 O 0 0 0 0 0 O 3

Non-reproducing 4 0 0 0 O O O O 0 0 4
Seedlings 0 17 0 0 0 0 0 0 0 8 25
Totals 5 19 0 O O O 0 0 0 8 32
1513 Reproducing 9 l 3 2 O O 0 0 O O 15
Non-reproducing 25 0 1 5 O 0 O 0 0 O 31
Seedlings 0 0 0 0 O 0 O 0 0 0 0
Totals 34 1 4 7 0 0 0 0 0 0 46
1516 Reproducing 3 0 0 O 0 2 O 3 3 O 15
Non-Reproducing 18 1 0 0 0 O O 9 0 O 28
Seedlings 2 0 0 O O O O 3 0 O 5
Totals 23 1 O 0 0 2 0 15 3 0 44
1593 Reproducing 11 O O 0 0 0 0 0 O 0 11
Non-Reproducing 27 4 0 O 0 0 O O 0 O 31
Seedlings 15 1 O O O O 0 0 0 O 16
Totals 51 5 O O O O O O O 0 56
1612 Reproducing 12 10 6 12 17 17 2 6 10 97
Non-Reproducing 19 6 2 13 16 8 1 2 2 7O
Seedlings 5 4 1 4 a 4 2 2 5 34
Totals 36 20 9 29 37 29 5 10 17 201

15

volcanic ash soils; fl. mexicanum was found on volcanic ash and rhyolitic
soils. Hieracium 'chipingue' and fl, 'potosi' were extensively sought
for in any place other than their known sites but no additional

localities were found.

Naturally occurring hybrids

A significant aspect of the summer field work was the search for
localities where species that are suspected of hybridizing were growing
side by side, and were still distinguishable. Virtually every species
of the polytypic group was found growing with every other species and
yet could still be fairly easily identified (Table 1). At one site in
Puebla, four different species were growing together and no hybrids were
found. This does not say that hybridization does not occur, it simply
indicates that conspicuous hybrid individuals or swarms were not
observed. If gene flow is occurring between species, other techniques,

such as gel electrophoresis, will be necessary to'document the matter.

Dispersion within a habitat

The results of the dispersion study (Table 2) clearly indicate
clumped occurence patterns. This seems to be due to two facts,
underground structure and seed dispersal mechanisms. The Hieracium
species investigated have a rhizomatous habit. Excavations of numerous
plants showed that new shoots were produced at the base of the old plant
once flowering had been completed. New shoots are produced along a
central rhizome and generally grow 1 to 10 cm away from the central root

prior to turning upward. While collecting live plants for breeding

16

system studies, it was common to gather 3 to 10 interconnected
individuals in a 10 cm square area.

Seed dispersal patterns also may lead to the clumped dispersion.
While running the transect, several individuals categorized as
"seedlings" were excavated and showed very tiny roots with no
underground connection to other members of the community, indicating
that they were indeed seedlings. In some cases these seedlings were
growing adjacent to a flowering individual, or even under the leaves of
mature plants. This indicates that some of the seed is not dispersed
very far by wind, despite the pappus. Stergios (1976) found this

pattern in Hieracium aurantiacum in Michigan, where most seed was

 

deposited within 1 meter of the source. Another consideration is the
fact that Hieracium species in Mexico are often on isolated mountains
which can be likened to islands of suitable habitat, surrounded by harsh
seas of arid or cultivated land. Shmida's (1985) work on Compositae
with an inconsistently deciduous pappus indicates that this condition is
of selective advantage in species that are found in habitat islands,
although it is not limited to those species. These findings expand
upon Carlquist's (1966A, 1966B, 1966C, 1967) work on seed dispersal
habits of Pacific Island Compositae and other groups, in which he found
many instances of island plants lacking the long distance seed dispersal

mechanisms which originally enabled them to colonize the island.

Locations in general

Although the areas where Hieracium are found are for the most part
not geologically recent (Jones 1976, Beaman and Andresen 1966, Beaman
1962, 1965; Martin and Harrell 1957, Dressler 1954, Sharp 1953), the

sites are recent in some respects. The polytypic species are almost

l7

always found in disturbed sites. They were not found in the most
recently disturbed situations; rather, they are a part of the wave of
secondary colonizers, occurring in the partial shade found under young
trees. This was amply illustrated by the search of the site of

Hieracium dysonymum collected by Rogers McVaugh in 1949 (McVaugh 9878),

 

/
a site in the state of Michoacan where they were reported as "abundant."

Three hours of searching yielded one individual (Soule 1393B) in 1986.
In a subsequent telephone conversation with Dr. McVaugh, he mentioned
that the plants were all over the slopes under "recently planted pines."
In 1987 the site was revisited with special attention paid to open areas

at the edges of the dense forest. Numerous fl, dysonymum were found

(Soule 1582, 1583).

 

Conclusions

 

Based on the fact that virtually every one of the species studied
was found growing along side every other species, with no clearly
intermediate individuals, it appears that hybridization may not be a
major factor in influencing variation.

Field work determined that the northern Sierra Madre Oriental

populations of Hieracium abscissum, and to a lesser degree 3. dysonymum

 

and fl, schultzii, are disjunct from those in the trans—Mexican volcanic

range. Hieracium pringlei has an almost continuous distribution.

 

Clumped dispersion and habitat preference indicate that the Mexican
Hieracium species are secondary colonizers which have suitable sites for
growth only for a short period of time prior to being forced to spread

to new locations by the closing of the forest canopy.

18

BREEDING SYSTEM STUDIES

Introduction

Naturally occurring hybrids have long been a thorn in the side of
the plant taxonomist (Wagner 1968; Baum 1969, 1971). Various types of
studies have been done in an effort to clarify the question of
hybridization in different groups as exemplified by the approaches of
Bell (1952), and Turner and Alston (1959). Summaries of various
approaches are found in the works of Constance (1964), Heiser (1963,
1966, 1969), and Turner (1964, 1967, 1969). One such approach is
breeding system studies which generally involve culturing the suspected
parental species and attempting to reproduce the hybrids (Jones 1966).
Breeding system studies have been done on a group of western North
American Hieracium species. Kruckeberg (1967) was able to cross two
different species that are found in the Olympic Mountains in Washington.
From these crosses he raised viable offspring, although the Ea
generation had reduced fecundity. Christy (1986) was able to cross
three species of eastern North American Hieracium but only two of the
crosses yielded viable offspring.

To determine if hybridization was a factor leading to the polytypy
of the Mexican species, a series of hybridization studies was planned.
Live plants were needed and they were obtained by field collection and

by growing seed that was field collected.

l9

Apomixis results in additional complex variation that affects plant
classification (Stebbins 1950). Localized populations that are
apomictic can vary slightly from other populations, resulting in a
tendency for prolific naming of new species. European species of
Hieracium are frequently apomictic and have fallen prey to rather
zealous workers with "...eyesight stimulated beyond that of the ancient
hawks...[resulting in]...thousands of so-called species, subspecies,
varieties and forms..." (Fernald 1950).

Based on preliminary observations, Beaman (personal communication)
suspected that some Mexican Hieracium species may be apomictic.
Cytological work in Hieracium dates back to Sears in 1917. Cytological
studies can prove polyploidy and apomixis as shown in the work on

Townsendia by Beaman (1954, 1957), and more recently in the work on Poa

 

by Kellogg (1987). Chromosome counts of Mexican species of Hieracium
have been done by various workers and the counts found only diploids
with g = 9 (Table 3). Other techniques used to determine apomicts in
Compositae include pollinator exclusion and emasculation. Emasculation
was done by Guppy (1978) in her work on British Columbian Hieracium.
Pollinator exclusion studies are useful in that they can show whether
the species is selfing and/or apomictic versus obligately outcrossing.
Christy (1986) demonstrated the technique's usefulness in her work on
three species of Hieracium in southeastern North America. Cytological
evidence, emasculation of flower heads, and pollinator exclusion studies

were utilized in this study.

20

Table 3. Chromosome counts of Mexican species of Hieracium.

 

 

 

 

 

 

 

 

Species Nufiber Reference

.fl. abscissum 9 McGregor 16584 (KANU), unpublished.

.fl. abscissum 9 Harms, 1973.

E, mexicanum 9 Beaman, DeJong, and Stoutamire, 1962.

‘E, mexicanum 9 Beaman and Turner, 1962.

E, pringlei 9 Keil, Luckow, and Pinkava, 1988.

.E. pringlei 9 Raven and Breedlove 19921 (MICH),
Raven, unpublished.

.fl. albiflorum 9 Stebbins, Jenkins, and Walters, 1953.

‘E, crepidispermum 9 Stebbins, Jenkins, and Walters, 1953.

g, crepidispermum 9 Powell and Turner, 1963.

E, gronovii 9 Jones, 1968.

E, irazuense 9 Beaman and Turner, 1962.

21

Methods

In the course of field work a search was made for any naturally
occurring hybrids. Other priorities included the collection of live
plants for breeding studies, the collection of seeds to grow into adult
plants for breeding studies, establishment of pollinator-exclusion
plots, and collection of pollen for pollen morphology studies.

In an effort to get field data on breeding systems, pollinator
exclusion plots were established. Two locations were selected. At the
first site, on Chipinque Mesa (near Monterrey, Nuevo Lean), ten flower
stalks were wrapped with cheese cloth to exclude pollinators. The
second site was at the base of Nevado de Toluca (near Toluca, State of
Mexico), where 20 flower stalks were wrapped with cheese cloth. Any
heads that were open, or that had completed flowering, were removed
prior to wrapping. The stalks were then tied to a metal stake to
prevent the stem from breaking when the cheese cloth became wet with the
summer rains. The metal stakes had yellow plastic flags on which a
request in Spanish was written "scientific investigation, please do not
remove."

In 1986 a total of 37 plants was field collected. In order to
bring the live material into the United States all soil was washed from
the roots, the roots wrapped in moist tissue, and the plants placed in
plastic bags with the appropriate collection numbers. These bags were
placed in a larger plastic bag which was sealed and refrigerated during
transit to Michigan State University where the plants were promptly
potted individually into 4" round clay pots and placed in a greenhouse.

The soil was a 9:1 mix of sterilized Bacto (a commercial mix of peat,

22

ground bark, and perlite) and coarse sand.

Three plants died within a few days from apparent heat stress. The
surviving plants were moved to a light bench indoors where they were
exposed to a 12—hour light regime with an intensity of 180 micro-
einsteins. The plants began flowering within three weeks of potting.

Emasculation by anther excision was done as outlined by Guppy
(1978) and Beaman (personal communication). All anthers in a head were
excised at once, with one stroke of a razor blade. A new razor blade
was used for each excision, and hands were washed between each excision.
These precautions were taken to avoid contamination of the successive
heads with pollen. Twenty heads in three species were utilized (Table 5).

Artificial self-pollination was attempted on flowers within the
same head and on flowers in different heads on the same plant.
Pollination of flowers within the same head was done by running a fine
paint brush across the head to move pollen to all stigmas. The paint
brush was swirled in 95% ethanol, washed with soap'and water, rinsed for
five minutes under running water and allowed to dry between uses.
Pollination of two heads on the same plant was done by rubbing heads
together. Two species and 18 heads were used for the self-pollination
study (Table 6).

A control group to the above studies was established by selecting
five plants which were not flowering at the same time as any other
plants, and hence could not be used for hybridization work. No
manipulation of flowers was done, including no wrapping to exclude
pollinators (Table 7).

In an effort to determine best growing conditions for Hieracium
prior to further collections of the study species, 12 plants of

Hieracium fendleri A. Gray ssp. fendleri were collected from the Santa

 

23

Rita Mountains of Pima County, Arizona in March 1987. Two were potted
in their own soil and the rest were potted into a gradient of
progressively more porous soil.

Two hundred forty-eight plants were field-collected in Mexico in
1987. They were immediately placed into 4" plastic pots in their own
soil. As the plants generally grow in clumps this often meant that
there were four to eight individuals per pot. The potted plants were
kept in a portable greenhouse in the back of the field vehicle. This
greenhouse was set outside whenever suitable conditions prevailed.

In order to bring the plants into the United States all soil had to
be removed from the roots according to U. S. Department of Agriculture
regulations. The majority of the soil was collected into paper bags for
treatment by methyl bromide for 24 hours at the inspection station. The
roots were washed to remove excess soil, wrapped in moist tissue, and
the plants placed in plastic bags with appropriate collection numbers.
These bags were sealed in larger plastic bags, wrapped in a towel to
avoid ice damage to the outer leaves, and placed in an ice-filled
cooler.

The plants remained in the cooler for five days prior to the return
to Michigan State University where they were potted individually into 4"
square plastic pots and placed on a growth bench indoors. Whenever
possible the plants were re-potted into the soil from their collection
site. When there was not enough native soil for all individual plants,
a 9:1 mix of sterilized Bacto and coarse sand was used. If there were
more than 10 individuals from a collection site, an additional soil mix
was used. This was a 9:1:2 mixture of Bacto, coarse sand, and

unsterilized soil from under the pines in the Beal Pinetum (Michigan

24

State University Campus). This last mix was used in an effort to
determine if the plant death of the previous year was due to a lack of
some mycorrhizal or other relationship with soil fungi found under
pines.

On the growth bench the plants were exposed to a 13-hour day length
and light intensity of 180 micro-einsteins. After four weeks, in an
effort to induce flowering, the day length was increased to a 14-hour
day, then, after an additional four weeks decreased to 12 hours. During
this time the plants were stricken by successive infestations of disease
and predatory insects. In sequential order the infestations and their
treatments: thrips treated with Temik; powdery mildew with Benlate;
white flies with Temik; fungus gnats (whose larvae eat soil fungus and
damage plant roots) were treated with a Benlate soil drench; mealy bug
treated with Malathion. Plants died at the rate of three to ten per
week. None of the 248 field—collected plants flowered. Several plants
sent up flower stalks, and had a few buds, but no buds opened, excluding
the opportunity to utilize the plants collected in 1987 in breeding
studies.

Seeds were field collected in 1986 and 1987. In some cases whole
heads had been harvested. Where possible, counts were made of the total
number of seeds (fruits) per head and the total number of viable seeds
per head (Table 8). These seeds were subsequently included in the
germination and growth studies outlined below.

Through trial and error a method for growing plants from seed was
determined. It was found that surface sterilization of seeds was
necessary due to the numerous fungi and bacteria present on the seed
coat. The seeds were surface sterilized by the following process which

started with a two—minute wash in 95% ethanol to remove waxes and

25

enhance imbibition of water. They were then rinsed three times with
distilled water, soaked for 8 hours in distilled water, then rinsed with
a 10% bleach solution. This insured that any fungal spores that had
germinated during the overnight soaking were killed. Three five-minute
rinses with distilled water were required to remove all traces of the
bleach solution. Seeds were then soaked in distilled water overnight
and rinsed again. Once the seeds were sterilized, all equipment was
surface sterilized and the remaining steps done in a laminar flow hood.
Seeds were placed for germination in sterile petri plates with MSO
nutrient agar solution as used in plant tissue culture. The plates were
placed in a growth chamber under constant environmental conditions.
Germinated seedlings were transferred into sterile MSO nutrient agar
media in boxes known as GA—7's or Magenta-7's, commonly used in tissue
culture. These were then kept in a growth chamber under constant
environmental conditions.

After approximately two months in the GA—7's, the nutrient media
became exhausted and the plants had to be transfered to fresh, sterile
GA—7's with a nutrient agar solution. In the transfer process some root
tips broke off. These tips were recovered for cytological work and
processed according to the procedures outlined by Radford et al. (1974),
but I was not able to find mitotically active cells.

The seeds collected in 1986 and grown in the GA—7's produced 74
individuals of a size suitable for flowering. However, all these plants
were killed by a growth chamber malfunction prior to flowering.

A limited amount of tissue culture was attempted. The apical
meristem and 4 to 10 leaf sections were isolated from five plants that

appeared to be dying. These were sterilized, sectioned, placed in GA-

26

7'5 with MSO media that included growth hormones. The GA-7's were then
placed in a growth chamber. The leaf tissue formed callus and three of
the five meristem sections showed new growth. A growth chamber
malfunction killed these tissues before significant results were

obtained.

Results and discussion

 

Pollinator exclusion

All marker flags at both field localities were removed by local
people during the period after they were installed and before my return
to the sites. In most cases the cheese cloth had been removed from the
plant and left on the ground, or was totally gone. Four of the ten
stalks were found still wrapped at the Chipinque Mesa site and six of
the 20 stalks were recovered from the Toluca site.

Total number of heads per stalk, total number of heads that were
sufficiently mature for seed set to be determined, and the number of
seeds set were recorded (Table 4). Heads were deemed mature enough for
seed set to be determined if they had shed their corollas and had a
conspicuous pappus. Generally it was easy to determine viable achenes
because they were filled out, plump, and in most cases, of a darker

color than those that were aborted.

Breeding Studies

Emasculation by anther excision was done on 20 heads of three
species (Table 5). There was a total of 3 viable seeds recovered. This
could indicate that the process of emasculation was so stressful to the

plant that it failed to set much seed. Conversely, the three seeds may

27

Table 4. Results of pollinator exclusion study.

 

 

 

 

 

# heads # heads old enough # seeds

Site & taxon per stalk to have seed (see text) recovered
Toluca, Mexico 8 1 O
_I_i_. mexicanum 4 1 O
(Soule 1593) 4 1 O
4 O -
4 l O
4 l 0
Total 0
Chipinque, lO 4 O
Nuevo Leon . 3
.fl. 'chipingue' . 7
(Soule 1696) . O
21 4 6
. O
. O
larvaei —
23 O -
7 3 O
. O
larvae* -
Total 16

*larvae indicates the larvae of Tephritid (Tephritidae) flies.
The flies lay their eggs in Compositae heads and the larvae
cause the formation of a protective gall as they eat the
developing seed.

28

Table 5. Results of anther excision (emasculation) in selected species
of Mexican Hieracium.

 

 

 

 

Soule
Collection Number # of seeds

Species Number of heads recovered
fl. crepidispermum 1398 3 O
O
. 0
Total 0
.H. abscissum 1407 6 + 1
Total 1
fl. mexicanum 1415 11 O
. 1
. O
. O
. O
. 1
. O
O 0
. 1
. O
. 0
Total 3

+ indicates that the heads opened in storage, exact counts per head
not possible.

29

Table 6. Results of artificial self-pollination in selected species

of Mexican Hieracium.

 

 

 

 

 

 

Soule
Collection Crossed Number of # of seeds
Species Number With replicates recovered
.H. pringlei 1424T other + 5 52
‘H. pringlei 1424W other + 2 13
.fi. pringlei 1424T same * 7 32
‘H. pringlei 1424T same * 1 15
.fl. mexicanum 1393 same * 1 11

other + indicates that the heads were crossed with other heads on
the same plant. '

same * indicates that the flowers in the heads were crossed with
other flowers in the same head (technique described in
text).

30

Table 7. Control group for breeding system studies in Mexican Hieracium,
heads left untouched and unwrapped.

 

 

 

Soule
Collection Number ! of seeds

Species Number of heads recovered
fl. abscissum 1396 12 + 1
""—“"’ "' 4 0
. 0
. 0
. 0
Total 1
.fl. mexicanum . 1392 7 0
. 0
. 0
. 0
. 0
. 0
. 0
Total 0
fl, mexicanum 1415 5 0
. 0
. 1
. 0
. 0
Total 1
fl, mexicanum 1419H 9 0
O 0
. 0
. 1
. 0
. 0
. l
. 0
O 0
Total 2
fl, mexicanum 14198 10 O
. 0
. 0
. O
. 1
. 2
. 1
. 0
. 1
, 0
Total 5

+ indicates that the heads opened in storage, exact counts per head
not possible.

31

Table 8. Counts of the number of seeds (fruits) per head in
field-collected heads.

 

 

 

 

Soule Soule
Collection Collection # of seeds
Species Site Number Number recovered
11. abscissum 4 1316 19
H. abscissum 4 1324 18
fl. abscissum - 4 _1_3_9_6 21
fl. abscissum 4 13_96 20
11. abscissum 4 13_96 25
11. abscissum 4 1326 18
fl. abscissum 4 1292 15
fl. abscissum 10 1491 28
fl, abscisswm 10 1491 10
Average 19.3
E, mexicanum 12 1414 15
11. mexicanum 13 _l_41_5_ 18
Average 16.5
11. dysonymum 11 1411 12
11. dysonymum 11 1m. 25
11. crepidispermum 8 1% 19
unidentified + SB none 15
unidentified + 5B none 20
unidentified + 11 none 18

+ unidentified indicates that the heads were collected from a
deceased plant. Positive identification not
possible based on material available.

32

have been the result of incomplete anther excision and subsequent
selfing, or they may have been due to pollination by white flies or
fungus gnats because the heads were not wrapped to exclude pollinators
after emasculation.

Artificial self-pollination was attempted on flowers within the
same head and on flowers in different heads on the same plant using two
species and 18 heads (Table 6). If these species are similar to
pseudogamous apomicts which require pollen tube growth to begin before
the female gametOphyte forms the embryo, then the fruit set should have
been higher, equivalent to the number set by plants in the field (Table 8).

A control group of five plants was established in which no
manipulation of flowers was done, including no wrapping to exclude any
pollinators (Table 7). The very low level of fruit set by these plants
suggests that they are not apomictic. The few fruits set may have been
caused by the actions of the fungus gnats or white flies which were
present.

The sparsity of test replicates and lack of interspecific crosses
were due to the constraints imposed by flowering periods. Hieracium
flowers open for only a short time, and although pollen can be collected
from flowers even after stigma receptivity has passed, the overlapping
time frame necessary failed to occur. Another confounding factor was
the fact that many buds aborted prior to opening, perhaps due to white
fly feeding. This conclusion is backed by the findings of Snow and
Stanton (1988) in which total fecundity of Raphanus was markedly reduced
due to aphid feeding. Overall, the plants were stressed in "captivity"
and died at the rate of one to three per week.

If one compares the results in Table 4 with those in Table 8, it

can be seen that unwrapped heads have a higher number of seed set.

33

Wrapped heads of Hieracium 'chipingue' yielded 16 seeds from 12 heads,
an average of 1.33 seeds per head, whereas fl, abscissum unwrapped

yielded an average of 19.3 seeds per head. Hieracium mexicanum had an

 

average number of 16.5 seeds per head unwrapped and 0 seeds per head
when wrapped. If the species are normally selfing, then these results
indicate some perturbation to the system due to the wrapping.
Otherwise, this demonstrates that outcrossing results in a greater
number of seeds than does selfing.

When the results of the pollinator exclusion study (Table 4) are
compared to the control group of the breeding studies (Table 7), an
interesting similarity is seen. The control group, in which the heads
were left alone and not wrapped, set virtually no seed as is the case in
the pollinator exclusion studies. If one were to argue that the control
group set no seed due to stressful environmental conditions, then it
would follow that the artifically selfed group (Table 6) would also set
no seed due to the same stressful environmental conditions. The
artifically self-pollinated plants had virtually the same number of
viable seed as the unwrapped plants in the native habitat (Table 8).
This indicates that the control group set few seed because ovules were

not fertilized and not because the plants were stressed.

Plants collected to test for optimal growth conditions

The Hieracium fendleri ssp. fendleri plants collected in Arizona in
March of 1987 all died quickly, although individuals potted in their own
soil survived almost three weeks longer than all others. This led to
the conclusion that Hieracium species should be collected, brought back,

and grown in their native soil whenever possible.

34

Conclusions

 

Overall, the data indicate that these Hieracium species are
outcrossers that will, in some cases, set seed after selfing. This work
does not find the plants apomictic. Further studies, under non-stressful

conditions for the plants, should be done.

35

VESTITURE STUDY

Introduction

 

One important morphological character for species determination in
Hieracium is the vestiture. Keys have been written based in
considerable part on types and densities of hairs. Vestiture also has
proven to be an aid to taxonomic classification in many other groups as
seen in the work of Hardin (1957), Solbrig (1960, 1961), Anderson
(1963), Webb and Carlquist (1964), Seith (1979), and Seith and Anderson
(1982).

Almost since its inception, scanning electron'microscopy (SEM) has
been used for the examination of vestiture, in part because of the great
depth of field, and in part due to the ease of preparation of samples
(Hammond and Mahlberg 1973). Numerous workers have used SEM
observations of vestiture as an aid to taxonomic classification,
including Baum and Hadland (1973, 1975), Rollins and Banerjee (1975,
1979C), Anderson (1983), Hardin (1986), Hansen (1986), Healey, Mehta and
Westerling (1986), Jones (1986), wurdack (1986), Hardin and Bell (1986),
Ehleringer and Cook (1987), Rosatti (1987), and Hannan (1988).

Preliminary work by Beaman with a dissecting microscope
(unpublished) has identified five major types of vestiture in Mexican

Hieracium:

1) tomentum at the base of the stems,

36

2) long pilose or hirsute vestiture on abaxial and adaxial
surfaces of leaves,
3) lanate pubescence on leaves and stems,
4) glandular hairs on phyllaries,
5) presence or absence of pubescence on the stem at the base of
the head.
This study investigated the vestiture at the base of stems, abaxial
and adaxial surfaces of the leaves, phyllaries of mature heads, and

stems at the base of heads.

Methods

Samples for the vestiture study were taken, with one exception,
from the 1986 and 1987 collections of J. A. Soule et al. The exception

was Hieracium pringlei, Rzedowski 30821 (MSC). All samples were from

 

air dried herbarium specimens and were not treated or rehydrated for
this study. Samples were selected from specimens based on three
criteria. 1) Individuals of each taxon were selected from the extremes
of the range, as well as from the center of the range; 2) plants that
had been growing with other taxa of Hieracium were preferentially
selected; 3) specimens were selected which included ample material so
that the specimen would not be left depauperate from the modest amount
of sample material removed.
Samples were taken from each of the following:
1) base of the stem (a 5 mm section immediately above the attachment of
the uppermost leaf in the basal rosette);

2) abaxial surface of the youngest mature leaf of the basal rosette (a

37

10 mm long section starting at petiolar insertion into the blade);
3) adaxial surface of the youngest mature leaf of the basal rosette (a

10 mm long section immediately adjacent to the previous sample);
4) phyllary of a mature head at anthesis or prior to seed set;

5) a 5 mm section of stem at the base of a mature head.

These samples were mounted on large size JEOL stubs (24 mm
diameter) using double stick tabs. Samples were prepared for viewing by
grounding with carbon paint and coating with gold in an EM-Scope SC 500
sputter coater. The best coat thickness is 35 nanometers. Less of a
coat, e.g. 21 nm, was not sufficient to eliminate charging.

A JEOL JSM—35CF scanning electron microscope located in the Center
for Electron Optics at Michigan State University was used to view the
samples. Settings were as follows: working distance 15, condenser lens
400, KV 15. Micrographs were taken using Polaroid Type 665
positive/negative film. A representative subset of all photographs
taken is presented, by taxa, in Figures 6 through 12.

Based on the photographs of the samples, the kinds of hairs were
defined following Payne (1978), and their presence or absence, and
relative length evaluated where applicable. The vestiture information
was quantified and these values, along with other key morphological
characters, were entered into a data matrix for evaluation by the MVSP
(MultiVariate Statistics Package) software developed by Kovach (1986).
Presence of a character was valued at 0.99 and absence was valued at
0.01. This technique for handling categorical data intended for
statistical analysis is used in the social sciences when the analysis
includes categorical and numerical data. The compiled raw data are

presented in Appendix II.

38

Due to the nature of the information gathered (a combination of
quantitative and qualitative values) the appropriate statistical test
was one that could take the two types of values into consideration and
still yield meaningful results. There are two such tests available on
the MVSP, Spearman's Rank Correlation and Average Linkage Cluster
Analysis. Both tests were utilized. A series of correlations was run,
first a combined correlation of all operational taxonomic units (OTUs)
at once, then a series comparing each species to each other species.
The Average Linkage Cluster Analysis was run on the same data groupings

as the Spearman's Rank Correlations.

Results and Discussion

 

Figures 6 through 12 demonstrate the typical vestiture found in
each species. Each figure contains six photographs. The first five
photographs, A through E, represent the five samples taken from each
specimen (see Methods). The sixth photograph in each figure is some
interesting feature observed in detail.

The vestiture consists of four types: epicuticular waxes, glandular
hairs, stellate hairs, and barbed hairs. Epicuticular waxes (see
example Figure 9-F) are present in all samples of all individuals of

Hieracium abscissum, H, pringlei, H, schultzii, and H, 'chipingue.’

 

They are not found at all on any of the sampless of H, dysonymum, H.
mexicanum, and fl. 'potosi.’ Thus these waxes can be used as a species-
specific character.

On the phyllary glandular hairs and stellate hairs are present in

all species. Barbed hairs are present only in Hieracium mexicanum and

39

.fl' schultzii. The phyllaries of all species show a rugose texture (see
example Figure 7-F).

On the stem below the head glandular hairs are present in most
samples. When present, the lengths of the glandular hairs were measured
All samples of Hieracium dysonymum and fl. 'potosi' lacked glandular
hairs on the stem below the head. This factor, along with the absence
of epicuticular waxes is a species-specific trait. Stellate hairs are
present in all samples. Barbed hairs are present only on H, mexicanum
and fl, schultzii.-

On the abaxial surface of the leaf glandular hairs are present in

Hieracium dysonymum, H. mexicanum, and fl. 'potosi.' Stellate hairs are

 

present in only two samples from two different species. Barbed hairs
are present in all species. When possible, length of the barbed hairs
was measured. The relative lengths of the barbs on the hairs were
evaluated as "long" or "short," long referring to those barbs mostly
longer than 40 micrometers, and short those between 10 and 35 micro-
meters. An example of a short barbs is found in Figure 6-E. This can
be compared to the long barbs which are found only in Hieracium
pringlei, Figure 9-E. This factor is a species-specific trait. The
presence or absence of glandular hairs on leaves was thought to be
species-specific. They were not found on the standard samples of
Hieracium abscissum; however, they were found on the sample of a very
young 5, abscissum leaf (Figure 6-F). The factthat the glands were not
found on mature leaves of the species may indicate that they are lost as
the leaf matures, or that this is an aberrant sample, or that they were
broken off during field collection of the plant.

On the adaxial surface of the leaf glandular hairs are present on

some samples. Stellate hairs are mostly lacking on lower leaf surfaces.

40

Figure 6. Scanning electron micrographs of the vestiture of Hieracium
abscissum.

A). Phyllary with waxes, glandular and stellate hairs.

B). Stem below the head with waxes, glandular and stellate hairs.
C). Upper surface of the leaf with waxes and short—barbed hairs.
D). Lower surface of the leaf with waxes and short-barbed hairs.
E). Base of the stem with short—barbed hairs; waxes lacking.

F). Upper surface of an immature leaf, note glandular hairs.

The bar on all photos indicates 100 micrometers.

Photos of samples taken from the following specimens:

A. Soule 1408 B. Soule 1573
C. Soule 1408 D. Soule 1648
E. Soule 1408 F. Soule 1408

 

41

 

Figure 6

42

Figure 7. Scanning electron micrographs of the vestiture of Hieracilun

A).

B).

C).

D).

E).

F).

The

dysonymum.

Phyllary with glandular and stellate hairs; waxes lacking.

Stem below the head with stellate hairs; waxes lacking.

Upper surface of the leaf with glandular and short—barbed hairs;
waxes lacking.

Lower surface of the leaf with glandular and short—barbed hairs;
waxes lacking.

Base of the stem with glandular and short-barbed hairs; waxes
lacking.

Phyllary surface, note rugose texture and lack of waxes.

bar on all photos indicates 100 micrometers.

Photos of samples taken form the following specimens:

A. Soule 1421 B. Soule 1421
C. Soule 1412 D. Soule 1421
E. Soule 1421 F. Soule 1412

 

 

 

 

 

 

 

43

 

Figure 7

44

Figure 8. Scanning electron micrographs of the vestiture of Hieracium

A).

B).

C).

D).

E).

F).

The

mexicanum.

Phyllary with glandular, stellate and short-barbed hairs; waxes
lacking.

Stem below the head with glandular, stellate and short—barbed hairms;
waxes lacking.

Upper surface of the leaf with glandular, stellate and short-barbed
hairs; waxes lacking.

Lower surface of the leaf with glandular, stellate and short-barbed

hairs; waxes lacking.

Base of the stem with glandular, stellate and short-barbed

hairs; waxes lacking.

/
Phyllary of specimen from Iztaccihuatl, above timberline. Note

density of short-barbed hairs.

bar on all photos indicates 100 micrometers.

Photos of samples taken from the following specimens:

A. Soule 1420 B. Soule 1420
C. Soule 1420 D. Soule 1420
E. Soule 1420 F. Soule 1614

 

 

 

 

45

 

Figure 8

46

Figure 9. Scanning electron micrographs of the vestiture of Hieracilnn

pringlei.

A). Phyllary with waxes, glandular and stellate hairs.

B). Stem below the head with waxes, glandular and stellate hairs.
C). Upper surface of the leaf with waxes, and long-barbed hairs.

D). Lower surface of the leaf with waxes, and long-barbed hairs.

E). Base of the stem long—barbed hairs; waxes lacking.

F). Stem below the head, note waxes and the base of a stellate hair.

Bar equals 1 micrometer.

The bar on all photos indicates 100 micrometers unless noted otherwise.

Photos of samples taken from the following specimens:

A. Soule 1579 B. Soule 1664
C. Soule 1664 D. Soule 1579
E. Soule 1664 F. Soule 1664

 

 

 

 

 

 

47

 

Figure 9

48

Figure 10. Scanning electron micrographs of the vestiture of Hieracium
schultzii.

A). Phyllary with waxes, stellate, glandular, and short-barbed hairs.

B). Stem below the head with waxes, stellate, glandular and short-barbed
hairs.

C). Upper surface of the leaf with waxes and short-barbed hairs.

D). Lower surface of the leaf with waxes and short—barbed hairs.

E). Base of the stem with waxes and short—barbed hairs.

F). Phyllary, note rugose texture, waxes and portion of a stellate hair.

Bar equals 10 micrometers.

The bar on all photos indicates 100 micrometers unless otherwise noted.

Photos of samples taken from the following specimens:

A. Soule 1409 B. Soule 1572
C. Soule 1549 D. Soule 1549
E. Soule 1409 F. Soule 1549

 

 

 

 

 

 

 

49

 

Figure 10

50

Figure 11. Scanning electron micrographs of the vestiture of Hieracium

'chipingue.'

A). Phyllary with waxes, glandular and stellate hairs.

B). Stem below the head with waxes, glandular and stellate hairs.

C). Upper surface of the leaf with waxes, and short—barbed hairs.

D). Lower surface of the leaf with waxes, and short-barbed hairs.

E). Base of the stem with waxes, and short-barbed hairs.

F). Detail of the stem below the head, note waxes on wrinkled epidermis,

stellate hairs and stomate. Bar equals 10 micrometers.

The bar on all photos indicates 100 micrometers unless otherwise noted.

Photos of samples taken from the following specimens:

A. Soule 1485 B. Soule 1485
C. Soule 1485 D. Soule 1485
E. Soule 1485 F. Soule 1485

 

 

 

 

 

 

 

51

 

Figure 11

52

Figure 12. Scanning electron micrographs of the vestiture of Hieracium

A).

B).

C).

D).

E).

F).

The

'potosi.‘

Phyllary with glandular and stellate hairs; waxes lacking.

Stem below the head with stellate hairs; waxes lacking.

Upper surface of the leaf with glandular and short-barbed hairs;
waxes lacking.

Lower surface of the leaf with glandular and short-barbed hairs;
waxes lacking.

Base of the stem with glandular and short—barbed hairs; waxes
lacking.

Cross section of stigma, broken at time of collection. Bar equals

10 micrometers.

bar on all photos indicates 100 micrometers unless otherwise noted.

Photos of samples taken from the following specimens:

A. Soule 1507 B. Soule 1507
C. Soule 1507 D. Soule 1507
E. Soule 1507 F. Soule 1507

 

 

 

 

 

 

 

53

 

Figure 12

 

56
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54

Barbed hairs are found in all species. The relative lengths of the
barbs on the hairs were also evaluated. Pubesence on the lower surface
of the leaf was commonly found on the mid-rib, as seen in Figures 7—D,
lO-D, ll-D, and 12-D. This may serve as a deterrent to insect herbivory
(Becerra and Ezcurra 1986).

On the stem at the base of the plant, glandular hairs are found

only in Hieracium dysonymum, fl, mexicanum, and fl. 'potosi.' Stellate

 

hairs are found on all samples of H. mexicanum and on no other samples
and can thus be considered a species-specific trait. Barbed hairs are
present on all species. The relative lengths of the barbs on the barbed
hairs were also evaluated. The presence or absence of epicuticular
waxes on these samples was frequently hard to determine as the stems had
often been handled extensively during collection.

The vestiture of an individual plant can vary over the course of a
season, depending on environmental conditions the individual is
subjected to, as shown by the work of Ehleringer and Cook (1987),
Ehleringer and Werk (1986) and Becerra and Ezcurra (1986). Therefore,
the vestiture data were recorded as length and presence or absence of a
particular type of hair, and the factor of density was not used in the
analysis.

The results of the vestiture study were evaluated by Spearman's
Rank Correlation using Kovach's (1986) MVSP program. The correlations
are presented in Tables 9-12.

Spearman's Rank Correlation gives an estimate of the similarity
between specimens, based on the values of the data input. This

similarity can be a positive or negative value. The closer the value is

to +1, the higher the correlation. For example, in Table 9, the value

55

1.00 appears when a specimen is compared to itself. The value 1.00 also
appears in two other places in Table 9 where two specimens are identical
as to characters surveyed.

One clear-cut example of the test results is seen with the

Hieracium mexicanum correlations; .H, mexicanum correlates generally

 

negatively with other species, although somewhat positively with
.H. dysonymum, and H. 'potosi.'

Table 9 yields a good indication of overall infrageneric grouping
in the species studied. If one adds the categories of 'headedness,‘
i.e. many heads per stalk versus few heads per stalk, and the pollen
character of straight versus curved spines (discussed in section on
pollen) there is a fairly clear-cut split. The many headed, curved-

spine pollen species are Hieracium abscissum, H. schultzii, and H.

 

'chipingue.’ These species are highly correlated with one another, and
lowly or negatively correlated with the remaining species. The
remaining group would be the few headed, straight'spined group which

includes Hieracium dysonymum, H, mexicanum, H, pringlei, and

 

H, 'potosi.’

Spearman's Rank Correlation was next done on the infrageneric
groups of many-headed or few-headed taxa. The comparison 0f.§°
abscissum, H. schultzii, and H, 'chipingue' (Table 10) indicates that
H, 'chipingue' is more closely allied to H. abscissum than it is to H.
schultzii. Note that in two places in Table 10, although all specimens
are from different populations, the value of 1.00 appears for different
collections of the same species. This indicates that the different
populations yield individuals that are virtually identical based on the
data surveyed. It would thus appear that plants with less highly

correlated values can indeed be thought of as separate species and not

56

 

 

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Table 10. Spearman's Rank Correlation Coefficients for the vestiture
data of Hieracium abscissum, H. schultzii, and H. 'chipingue.'

 

 

absl408 abslS73 absl648 sch1409 sch1549 sch1572 chi1485

absl408 1

ab81573 0.88 1

absl648 0.88 1 1

sch1409 0.75 0.87 0.87 1

sch1549 0.74 0.87 0.87 1.00 1

sch1572 0.81 0.93 0.93 0.94 0.94 1

chi1485 0.81 0.92 0.92 0.84 0.84 0.89 1

Table 11. Spearman's Rank Correlation Coefficients for the vestiture
data of Hieracium dysonymum, H, pringlei, and H. 'potosi.‘

 

 

dysl412 dysl421 dyslSSO prilS75 pri1664 pr30821 nu11519 nu11523 pot1507

dysl412 1

dysl421 0.94 l

dyslSSO 0.63 0.57 1

prilS75 0.37 0.30 0.71 1

pri1664 0.40 0.32 0.73 0.94 1

pr30821 0.30 0.22 0.62 0.86 0.81 1

nu11519 0.41 0.35 0.49 0.59 0.60 0.74 1

nu11523 0.41 0.35 0.49 0.59 0.60 0.74 1 1

pot1507 0.70 0.63 0.29 -0.01 0.02 -0.08 0.06 0.06 1

Table 12. Spearman's Rank Correlation Coefficients for the vestiture
data of Hieracium dysonymum, H. mexicanum, and H, 'potosi.‘

 

 

dysl412 dysl421 dy51550 mex1420 mex1593 mex1614 pot1507

dysl412 1

dysl421 0.94 1

dy51550 0.63 0.57 1

mex1420 0.45 0.51 O 09 1

mex1593 0.44 0.37 0.23 0.76 1

mex1614 0.31 0.26 0.24 0 66 0.90 1

pot1507 0 70 0.63 0 29 0 58 0.58 '0.44 1

58

 

 

Table 13. Average Linkage Cluster Analysis of vestiture data of all
taxa of Hieracium in the study.
AVERAGE NUMBER OF OBJECTS
NODE GROUP 1 GROUP 2 SIMILARITY IN FUSED GROUP
1 abs1573 absl648 1.00 2
2 nu11519 nu11523 1.00 2
3 sch1409 sch1549 1.00 2
4 dysl412 dysl421 0.94 2
5 pri1575 pri1664 0.94 2
6 NODE 3 sch1572 0.94 3
7 NODE 1 chi1485 0.92 3
8 mex1593 mex1614 0.90 2
9 NODE 7 NODE 6 0.88 6
10 NODE 5 pr30821 0.83 3
11 absl408 NODE 9 0.81 7
12 NODE 11 dyslSSO 0.77 8
13 mex1420 NODE 8 0.71 3
14 NODE 4 pot1507 0.66 3
15 NODE 12 NODE 10 0.65 11
16 NODE 15 NODE 2 0.58 13
17 NODE 14 NODE 13 0.44 6
18 NODE 16 NODE 17 0.15 19

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63

just different populations.

The correlations between Hieracium dysogymum, fl, pringlei. and fl.
'pgtggif (Table 11) show that g, 'potosi' is, in part, negatively
correlated to g, pringlei; fl, pringlei is also fairly lowly correlated
to E. dysonymum. Combined, this indicates that g. ‘potosi' is not very
likely to be a hybrid of E, dysonymum with E, pringlei.

Hieracium 'potosi' is more closely correlated to g, mexicanum than

 

it is to E. pringlei (Table 12). .fl. 'potosi' is more highly correlated
t°.fl° dysonymum than it is to g, mexicanum, except for the g, dysonymum

population that is closest to it in range (Soule 1550). This individual

 

also does not correlate highly with the other two samples of its
species. If one averages the correlations of g, 'potosi' with E.
dysonymum, and fl. 'potosi' with E, mexicanum, they average to 54.0 and

53.3 respectively. However, based on the fact that E, dysonymum (Soule

 

1559) is so slightly correlated with other specimens of its species, and
eliminating that value from the averaging, one gets an average
correlation 0f.§° 'potosi' with E. dysonymum, and fl. 'potosi' with E.
mexicanum of 66.5 and 53.3 respectively. This suggests that E. 'potosi'
is more closely allied to E. dysonymum than to E. mexicanum.

The results of the Spearman's Rank Correlation were next evaluated
by Average Linkage Cluster Analysis using Kovach's (1986) MVSP program.
The Cluster Analysis results are presented in Tables 13 through 16.

This information was then utilized to formulate the dendrograms
presented in Figures 13 through 16. The lengths of the branches in the
dendrograms are taken from the reported average similarity values.

Table 13 and Figure 13 show the results of Cluster Analysis of all

taxa in this study combined. Note that Hieracium 'chipinque' is more

 

64

similar to E, abscissum than 3. 'potosi' is to E. dysonymum. Based on
the samples selected, a. pringlei, fl. mexicanum, and fl. dysonymum all
show a higher degree of polytypy than do fl. schultzi or g. abscissum;
indeed, fl. 'chipingue' is more similar to _I_{_. abscissum than the E.
mexicanum samples are to one another.

Table 14 and Figure 14 show the results of Cluster Analysis of

Hieracium abscissum, fl. schultzii and fl. 'chipingue.’ This is the same

 

as the information presented in Table 13 and Figure 13, yet it is easier
to note the fact that E. 'chipingue' is more closely linked to two of

the g. abscissum collections (Soule 1573, 1648) than they are to another

 

.fl. abscissum sample (Soule 1408).
Table 15 and Figure 15 show the results of Cluster Analysis of

Hieracium dysonymum, fl. pringlei, and fl. 'potosi.’ The length of the

 

connecting branches becomes longer but overall the clusters are
equivalent to those in Table 13 and Figure 13.
Table 16 and Figure 16 show the results of Cluster Analysis of

Hieracium dysonymum, fl, mexicanum, and fl. 'potosi.’ By comparing

 

Figure 15 to Figure 16 it is once again indicated that Hieracium
dysonymum and fl. 'potosi' are more closely linked to one another than
they are to E. mexicanum or E, pringlei. The comparison of the two
dendrograms also indicates that E} mexicanum is more closely related to

.E. dysonymum/'potosi' than is E. pringlei.

Conclusions

 

Evaluation of vestiture data using Spearman's Rank Correlation and
Average Linkage Cluster Analysis indicates that there are two major

groupings of the Hieracium species studied here. One group is the

65

Hieracium abscissumjfl, schultzii group to which E. 'chipingue' belongs;
.fl. 'chipingue' is also shown to be closely related to E. abscissum,
indeed, in scanning electron micrographs they are impossible to tell
apart. The other group is the Hieracium dysonymum-mexicanum group to

which E. 'potosi' belongs. Hieracium 'potosi' is shown to be more

 

closely allied to g, dysonymum than to E, mexicanum, but not as closely
linked as g. 'chipingue' is to E, abscissum. Based on this evidence

‘fl. 'chipingue' should either be submerged into E, abscissum, or should
be given a low, perhaps varietal level ranking. Hieracium lpotosi' is
distinct enough from E. dysonymum to be given a higher level ranking,
variety if g. 'chipingue' is submerged, and subspecies if it is not.

Hieracium schultzii is determined to be segregated from, though

 

closely related 5} abscissum. Hieracium pringlei is a highly polytypic

 

taxon yet it maintains a clear-cut species identity.

66

POLLEN STUDIES

Introduction

 

Pollen grains have been utilized as an aid to clarifying the
systematics of numerous groups. WOdehouse did a series of
investigations into Compositae pollen using light microscopy, beginning
in 1926 and running through 1945, and could be viewed as one of the
founding fathers of this line of research. He found it possible to
trace phylogenetic trends in the Compositae based on palynological
evidence.

In early palynological work, artists' renditions were used to
record investigations (i.e. Nodehouse 1930). Later, photographs were
used but, due to the very narrow depth of field in light microscopy,
most images appeared at least partially blurry. Often artists'
renditions were used to try to clarify the photographs, as seen in the
work of Carlquist (1961) and Canright (1953). Workers often used
overall pollen size to make species distinctions since fine surface
detail is difficult to view or evaluate with light microscopy (Stone
1963; Kapadia and Gould 1964; Buchholz and Kaeiser 1940).

The advent of scanning electron microscopy (SEM) revolutionized
pollen viewing by providing a greater depth—of-field than light
microscopy, resulting in a very sharp, virtually three—dimensional

photographic image, and eliminating the vagrancies of recording by a

67

non-artist's hand (Martin 1969). Furthermore, the resolution in SEM is
such that surface features not visible with light microscopy can be
resolved and accurately measured. The relative ease of preparation is
an added attraction to its use (Martin and Drew 1969, 1970; Hanks and
Fairbrothers 1970).

The SEM has been used to survey the pollen grains of many groups
and has proved a valuable aid in systematic studies (Skvarla and Turner
1966; Lewis 1977). Workers have found that measurements of various
external features such as length of spines, lacunae and annulus
features, and texture and patterning of the exine of the pollen grains
can often clearly differentiate species. A partial list of taxonomic
work based in part on SEM of pollen includes the work of M. Takahashi
(1987), H. Takahashi (1986), Elisens (1986), Terrell et al. (1986),
Donoghue (1985), vanWyk and Dedekind (1985), Constance and Chuang
(1982), Rollins and Banerjee (1979A and 1979B), Keeley and Jones (1977),
Anderson and Gensel (1976), Baum et al. (1970, 1971), Jones (1970). Of
particular note is the work by Baum, Bassett and Crompton (1971) which
combined light microscopy with SEM, and resulted in a key to 52 species
of Tamarix based on pollen morphology. Of greater significance to the
work here is Jones' 1970 study of Vernonia (Compositae). Jones combined
scanning electron micrographs with hybridization experiments and
cytological evidence to clarify relationships in the genus.

SEM examination of pollen was conducted to determine species
affiliation in the Hieracium taxa involved in this study. Pollen was
field-collected from all taxa except fl, schultzii which was collected

from unopened anthers of a dried specimen.

68

Methods

Pollen was field-collected for SEM observation by carrying the SEM
stubs into the field and applying double-stick tabs to the stubs
immediately prior to pressing them into the open florets of Hieracium
heads. Pollen was obtained at 24 collection sites. Voucher specimens
were collected once the pollen sample was taken. All collections are
those of J. A. Soule and J. P. Bodeau in 1987. Voucher specimens are in
the Beal-Darlington Herbarium, Michigan State University (MSC).

Pollen stubs were prepared for viewing by sputter—coating with gold
in an EM-Scope SC 500 sputter coater. Three coating thicknesses were
tried, 21 nm, 14 nm, and 10.5 nm. The 14 nm coat resulted in the best
compromise between absolute veiwing clarity and lack of charging.

A JEOL JSM—35CF scanning electron microscope at the Center for
Electron Optics at Michigan State University was used to view the
pollen. Settings were as follows: working distance 15; condenser lens
400; KV 15. Micrographs were taken using Polaroid Type 665
positive/negative film. Figures 17 and 18 contain a representative
sample of the photographs obtained.

A series of measurements was taken: overall grain diameter, spine
height, spine height above the pits, and number of spines on the annulus
and lacunae. The diameter was calculated by placing a piece of tracing
paper over the photograph and tracing the outermost edges of the pollen,
including spines. The diameters were then calculated from the circle
created. Total length of the spines located on the annulus (around the
germinal pore) was measured. A flexible ruler was used so that in the

case of curved spines the entire length could be evaluated. An

69

additional measurement of the length of spines above the pits was taken
and the percentage of spine above the pits determined. The pits
characteristically occur on only the lower half of the spine. In cases
where the spines on the annulus were not visible, spines surrounding the
lacunae were used. Results of these measurements are in Table 17.

A pollen character that seemed to vary between taxa was the number
of spines on the annulus and around the lacuna. The values vary between
8 and 12, and are always even numbers. Counts were taken in as many
instances as were possible.

It should be mentioned that all pollen work was done "blind," that
is without identifying the species from which the pollen was collected

until final tabulation of results.

Results and Discussion

 

Based on external pollen morphology, it is easy to separate the
Mexican species of Hieracium investigated into two groups, the "curved-
spine" group and the "straight-spine" group as seen in Figure 17; on the
left are the straight-spine group, Hieracium dysonymum, fl. mexicanum,
and fl, 'potosi' and on the right are the curved—spine group,

H, abscissum, H, schultzii, and H, 'chipingue.’ The two groups actually
correspond well with the plants' growth form. The many headed species

' have curved spines;

-- fl, abscissum, H. schultzii, and fl. 'chipingue.
the few headed species -- fl. dysonymum, H. mexicanum, and H. 'potosi,‘
have straight spines.

Hieracium pollen is spherical, triporate, echinate with

scrobiculate (pitted) spines, and has a lacuna immediately adjacent to

70

Figure 17. Field—collected Hieracium pollen.

A). H, dysonymum B). H. abscissum
C). H, mexicanum D). H, schultzii
E). H, 'potosi' F). H, 'chipingue'

The bar on all photos indicates 10 micrometers.

Voucher specimens for samples:

A. Soule 1586 B. Soule 1573
C. Soule 1593 D. Soule 1409
E. Soule 1507 F. Soule 1485

 

 

 

 

7l

 

Figure 17

Figure 18. Field—collected Hieracium pollen which has germinated.
A). H, abscissum B). H. dysonymum

C). H, mexicanum , D). H. pringlei

E)'.fl° 'chipingue' F)°.fl° 'potosi'

The bar on all photos indicates 10 micrometers.

Voucher specimens for samples:

A. Soule 1654 B. Soule 1586

 

C. Soule 161
E. Soule 148

. Soule 1516
Soule

N

 

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0

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U1
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Figure 18

74

Table 17. Evaluations of species of Mexican Hieracium pollen grains
based on scanning electron micrographs. All measurements
in micrometers.

 

 

 

 

 

 

Soule # of # of spine overall length I spine
Coll. Dia- spines/ spines/ geo- spine above above
Species number meter pore lacuna metry length pits pits
.H. abscissum 1573 23.636 10 - c 5.455 2.727
“H. abscissum 1573 -- 10 -- c 8.182 4.091
H, abscissum 1559 25.455 -- - c —-- --
'H. abscissum 1559 26.364 - 12 c -- --
H, abscissum 1559 25.455 -- 12 c -- --
.H. abscissum 1654 21.000 - - c -- --
H, abscissum 1654 21.667 -- 12 c ——- -—
Average 23.929 10 12 c 6.819 3.409
S.E. 0.901
H, dysonymum 1412 28.333 - - s -- -—
H, dysonymum 1412 27.500 -- - s —-- —--
.H. dysonymum 1550 24.000 10 8 5 4.000 2.000
H, dysonymum 1586 22.667 -- 8 5 ~-- --
'H. dysonymum 1586 27.778 -- 8 5 4.444 2.222
Average 26.056 10 8 3 4.222 2.111
S.E. 1.14
H, mexicanum 1593 -- -- 8 s -- ---
H. mexicanum 1593 -- -- 8 s -- ——-
E. —_mexicanum —1593 23 .500 12 8 3 4.000 2.000
H, mexicanum 1594 -- -- -— 5 5.556 3.333
H, mexicanum 1604 30.000 12 10 3 5.000 2.222
H, mexicanum 1604 -—- -- -- 5 5.556 3.333
.H. mexicanum 1605 29.333 12 10 3 4.667 2.667
H, mexicanum 1605 —- 12 10 s -- —--
H, mexicanum 1605 27.220 -- - 5 5.556 2.778
H, mexicanum 1612 29.333 12 10 3 5.333 2.667
H, mexicanum 1612 25.556 -- -— 5 4.444 2.667
H, mexicanum 1612 23.889 -- - s —-- -—
.H. mexicanum 1629 26.111 -- 10 5 5.000 3.333
Average 26.867 12 10 or 8 5 5.012 2.667

S.E. 0.892

75

Table 17 (continued). Evaluations of species of Mexican Hieracium
pollen grains based on scanning electron
micrographs. All measurements in
micrometers.

 

 

 

 

 

 

 

 

 

 

 

Soule # of # of spine overall length Z spine
Coll. Dia- spines/ spines/ geo- spine above above
Species number meter pore lacuna metry length pits pits
H, pringlei 1516 25.00 10 8 s -—- —- --—
.H, pringlei 1516 27.50 -- —— s -— -- --—
‘H, pringlei 1516 22.73 - 8 3 4.545 2.045 44.99
‘H, pringlei 1516 -- 10 8 5 6.000 2.000 33.33
‘H, pringlei 1519 24.06 - 8 s -- -- --
H, pringlei 1579 25.00 10 8 3 4.000 2.000 50.00
Average 24.86 10 8 3 5.136 2.136 42.50
S.E. 0.783
H, schultzii 1409 25.56 12 - c 5.56 3.33 59.98
H, schultzii 1409 27.22 -— —- c , —-- ——- --
Average 26.66 12 - c 5.56 3.33 59.98
S.E. 1.103
H, 'chipinque' 1696 26.36 10 -- c 5.45 3.64 66.79
.H. 'chipingpe' 1696’ 28.18 10 12 c 3.64 2.73 75.00
.H. 'chipinque' 1696 26.36 -- 12 c 3.64 2.73 75.00
.H. 'chipinque' 1696 29.44 10 12 c --- -—- --
H, 'chipinque' 1696 25.00 10 12 c 4.00 2.00 50.00
Average 27.07 10 12 c 4.20 2.78 66.70
0.778
H, :potosi: 1507 28.000 -- 8 5 4.666 2.000 42.91
H, 'potosi' 1507 -- -- 8 s --- —-— ---
H, 'potosi' 1513 26.154 10 -- 5 3.846 -- ---
.H, 'potosi' 1513 28.421 -- 8 s -—- -—- ---
H, pot031 1513 25.000 10 8 s -- --- —-—
Average 26.592 10 8 ‘ 5 4.256 2.000 42.91

S.E. 0.6896

76

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77

each germinal pore. Terms used are defined by Erdtman (1954). Overall
grain diameter was recorded. The average and standard error for the
diameter of each taxon was determined (Table 17 and Figure 19).

In general overall pollen diameter cannot be used to distinguish
species in this group. The differences in pollen size between members
of a single species may be due to differential energy expenditure by the
pollen parent as demonstrated in the work by Bell (1959), or it may
represent an actual taxon-related character. A larger sample size is
needed to make a definitive statement.

Total length of the spines and the percentage of spine above the
pits was determined. The spines are pitted on the lower 42 to 60
percent of the spine. The amount of extension of the spine above the
pits could not be used to distinguish species. It should be mentioned
that the presence of such prominent spines is indicative of an insect
pollinated as opposed to a wind pollinated group (Grashoff and Beaman
1970).

The most taxon-specific character was the number of spines on the
annulus and around the lacuna. Hieracium abscissum consistently had 10
spines per annulus and 12 per lacuna as did H. 'chipingue.’ Hieracium
dysonymum and H. [pggggi' had 10 and 8 as did H. pringlei. Hieracium
mexicanum spines numbered 12 and 10 or 8, and H. schultzii had 12
annular spines.

A surprising discovery was that germinated pollen was found on all
stubs to which stigmatic tissue had adhered (Figure 18). Stubs lacking
stigmatic tissue had no germinated pollen. Although there can be some
criticism of utilizing field-collected pollen (possible contamination by
pollen of other species), the unexpected discovery of germinated pollen

provided the benefit of knowing that at least some of the pollen

78

was viable.

Observation showed that on almost all stubs no malformed,
collapsed, or significantly smaller pollen grains were observed. This
fact combined with the evidence of viable, germinated pollen suggest
that the species examined in this study are not apomictic. The light
microscopy studies of Beaman (1957) and Grashoff (1969) on other
apomictic New World Compositae found a high number of malformed,
collapsed, or significantly smaller pollen grains. One might expect to
find this result in Mexican Hieracium as well. It might also be
reasonable to expect that one would not find a proliferation of

germinated pollen in the case of apomicts.

Conclusions

 

The Mexican species of Hieracium investigated can be separated into
two groups based on pollen morphology, the "curved-spine" group and the
"straight-spine" group, as seen in Figure 17. The two groups correspond
with growth form. The many headed species - Hieracium abscissum,

I

H, schultzii, and H. 'chipingue, have curved spines; the few headed

species -- H. dysonymum, H, mexicanum, H, pringlei, and_H. 'potosi,’
have straight spines.

The number of annular and lacunar spines proved species constant
and provides evidence for not recognizing Hieracium 'chipinque' and
.H. 'potosi' as separate taxa since these pollen characters correspond

to those of H. abscissum and H. dysonymum respectively.

BIBLIOGRAPHY

79

BIBLIOGRAPHY

Anderson, G.J. and P.G. Gensel. 1976. Pollen morphology and
systematics of Solanum sect. Basarthrum. Pollen et Spores
18:533-552.

 

Anderson, L.C. 1963. Studies on Petradoria (Compositae): Anatomy,
cytology, taxonomy. Kansas Acad. Sci. 66:632-684.

 

Anderson, L.C. 1983. Trichomes and stomata of Gordonia lasianthus
(Theaceae). Sida 10:103-113.

 

Baum, B.R. 1969. On the application of nomenclature to the taxonomy
of hybrids. Taxon 18:670-671.

Baum, B.R. 1971. The taxonomic and cytogenetic implications of the
problem of naming amphiploids of Triticum and Secale. Euphytica
20:302-306.

Baum, B.R. and V.E. Hadland. 1973. A scanning electron microscopic
study of epicuticular waxes of glumes in Avena magan, A, murphyi,
and A. sterilis. Canad. J. Bot. 51:2381-2383.

 

Baum, B.R. and V.E. Hadland. 1975. The epicuticular waxes of glumes
of Avena: a scanning electron microscopic study of the morphological
patterns in all the species. Canad. J. Bot. 53:1712-1718.

Baum, B.R., I.J. Bassett, and C.W. Crompton. 1970. Pollen morphology
and its relationships to taxonomy and distribution of Tamarix,
series Vaginantes. Osterr. Bot. Z. 118:182-188.

 

Baum, B.R., I.J. Bassett, and C.W. Crompton. 1971. Pollen morphology
of Tamarix species and its relationship to the taxonomy of the
genus. Pollen et Spores 13:497-521.

Beaman, J.H. 1954. Chromosome numbers, apomixis, and interspecific
hybridization in the genus Townsendia. Madrofio 12:169-180.

 

Beaman, J.H. 1957. The systematics and evolution of Townsendia
(Compositae). Contr. Gray Herb. No. 183.

Beaman, J.H. 1962. The timberlines of Iztaccihuatl and Popocatepetl,
Mexico. Ecology 43:377-385.

80

Beaman, J.H. 1965. A preliminary ecological study of the alpine flora
of Popocatepetl and Iztaccihuatl. Bol. Soc. Bot. Mexico 29:63-75.

Beaman, J.H. and B.L. Turner. 1962. Chromosome numbers in Mexican and
Guatamalean Compositae. Rhodora 64:271-276.

Beaman, J.H. and J.W. Andresen. 1966. The vegetation, floristics and
phytogeography of the summit of Cerro Potosi, Mexico. Amer. Midl.
Naturalist 75:1-33.

Beaman, J.H., D.C.D. DeJong, and W.P. Stoutamire. 1962. Chromosome
studies in the alpine and subalpine floras of Mexico and Guatemala.

Becerra, J. and E. Ezcurra. 1986. Glandular hairs in the Arbutus
xalapensis complex in relation to herbivory. Amer. J. Bot.

73:1427-1430.

Bell, C.R. 1952. Natural hybrids in the genus Sarracenia. I. History,
distribution and taxonomy. J. Elisha Mitchell Sci. Soc. 68:55-80.

 

Bell, C. R. 1959. Mineral nutrition and flower to flower pollen size
variation. Amer. J. Bot. 46:621—624.

Braun, B.L. 1955. The phytogeography of unglaciated eastern United
States and its interpretation. Botan. Rev. 21:297—375.

Buchholz, J.T. and M. Kaeiser. 1940. A statistical study of two
variables in the Sequoias——pollen grain size and cotyledon number.
Amer. Naturalist 74:279—283.

Canright, J.E. 1953. The comparative morphology and relationships of
the Magnoliaceae--II. Significance of the pollen. Phytomorphology
3:355-365.

 

Carlquist, S. 1961. Pollen morphology of Rapateaceae. Aliso 5:39-66.

Carlquist, S. 1966A. The biota of long-distance dispersal. II. Loss
of dispersibility in Pacific Compositae. Evolution 20:30-48.

Carlquist, S. 19668. The biota of long-distance dispersal. III. Loss
of dispersibility in the Hawaiian flora. Brittonia 18:310—335.

Carlquist, S. 1966C. The biota of long-distance dispersal. IV. Genetic
systems in the floras of oceanic islands. Evolution 20:433-455.

Carlquist, S. 1967. The biota of long-distance dispersal. V. Plant
dispersal to Pacific islands. Bull. Torrey Bot. Club 94:129-162.

Christy, C.M. 1986. A biosystematic study of the relationships of
Hieracium gronovii L., H. venosum L., and H. traillii Greene
(Compositae: Lactuceae). Unpublished Masters Thesis; University of
Tennessee, Knoxville.

 

81

Clausen, J., D.D. Keck and W.M. Hiesey. 1941. Regional differentiation
in plant species. Amer. Naturalist 75:231-250.

Constance, L. 1964. Systematic Botany - An unending synthesis.
Taxon 13:257-273.

Constance, L. and T.I. Chuang. 1982. SEM survey of pollen morphology
and classification in Hydrophyllaceae (Waterleaf Family). Amer. J.

Donoghue, M. J. 1985. Pollen diversity and exine evolution in Viburnum
and the Caprifoliaceae sensu lato. J. Arnold Arbor. 66:421-469.

Dressler, R.L. 1954. Some floristic relationships between Mexico and
the United States. Rhodora 56:81-96.

Ehleringer, J.R. and C.S. Cook. 1987. Leaf hairs in Encelia
(Asteraceae). Amer. J. Bot. 74:1532-1540.

Ehleringer, J.R. and K.S. Werk. 1986. Modifications of solar-radiation
absorption patterns and implications for carbon gain at the leaf
level. Pp. 57-82, in, T.J. Givnish, ed., On the Economy of Plant
Form and Function. Cambridge University Press.

Elisens, W.J. 1986. Pollen morphology and systematic relationships
among new world species in tribe Antirrhineae (Scrophulariaceae).
Amer. J. Bot. 73:1298-1311.

 

Erdtman, G. 1954. An introduction to pollen analysis. Chronica
Botanica Co., Waltham, Massachusetts.

Fernald, M.L. 1950. Gray's manual of botany. 8th edition, American
Book Co., New York.

Flores, H. N. 1985. La vegetacion del Cofre de Perote, Veracruz,
Mexico. Biotica 10:35-64.

Gleason, H.A. 1952. Illustrated flora of Northeastern United States
and adjacent Canada. Volume 3, New York Botanical Garden; Hafner
Publ. Co. Inc., New York.

Grashoff, J.L. 1969. Reproductive processes affecting the taxonomy of
some Mexican and Central American species of Eupatorium.
Unpublished Masters Thesis; Michigan State University, East Lansing.

Grashoff, J. L. and J. H. Beaman. 1970. Studies in Eupatorium
(Compositae). III. Apparent wind pollination. Brittonia 22:77-84.

Grashoff, J. L., M.W. Bierner and D.K. Northington. 1972. Chromosome
numbers in North and Central American Compositae. Brittonia
22:379-394

82

Guppy, G.A. 1978. Species relationships of Hieracium (Asteraceae) in
British Columbia. Canad. J. Bot. 56:3008-3019.

Hammond, C. T. and P. G. Mahlberg. 1973. Morphology of glandular hairs
of Cannabis sativa from scanning electron microscopy. Amer. J. Bot.
60:524-528.

 

Hanks, S. and D. E. Fairbrothers. 1970. Effects of preparation
technique on pollen prepared for SEM observations. Taxon 19:879-886.

Hannan, G. L. 1988. Evaluation of relationships within Eriodictyon
(Hydrophyllaceae) using trichome characteristics. Amer. J. Bot.
75:579-588.

 

Hansen, C. 1986. Notes on some foliar glands mainly in Asiatic genera
of the Melastomataceae. Bot. Jahrb. 106:15-22.

Hardin, J. W. 1957. A revision of the American Hippocastanaceae.
Brittonia 9:145-195.

Hardin, J. W. and J. M. Bell. 1986. Atlas of foliar surface features
in woody plants. IX. Betulaceae of eastern United States.
Brittonia 38:133-144.

Harms, L.J. 1973. Doccument of plant chromosome numbers. Sida

Healey, P.L., I.J. Mehta, and K.E. Westerling. 1986. Leaf trichomes of
some Parthenium species. Amer. J. Bot. 73:1093-1099.

 

Heiser, C.B., Jr. 1963. IV. Modern species concepts: Vascular
plants. Bryologist 66:120-124.

Heiser, C.B., Jr. 1966. Methods in systematic research. BioScience

Heiser, C.B., Jr. 1969. Taxonomy. In, J. Ewan, ed., A short history
of botany in the United States. Hafner Publ. Co., New York.

Holmgren P.K., W. Keuken, and E.K. Schofield. 1981. Index Herbariorum.
Part I, Edition 7. Gen. Ed. F. A. Stafleu.‘ Dr. W. Junk B.V., Publ,
The Hague, Boston.

Jardel, E. J. 1986. Efecto de la explotacion forestal en la estructura
y regeneracion del bosque de coniferas de la vertiente oriental del
Cofre de Perote, Veracruz, Mexico. Biotica 11:247-270.

Jones, J.H. 1986. Evolution of the Fagaceae: The implications of
foliar features. Ann. Missouri Bot. Gard. 73:228-275.

Jones, 8.8. 1966. Experimental hybridizations in Vernonia
(Compositae). Brittonia 18:39-44.

83

Jones, S.B. 1968. Chromosome numbers in Southeastern Compositae.
Bull. Torrey Bot. Club 95:393-395.

Jones, S.B. 1970. Scanning electron microscopy of pollen as an aid to
the systematics of Vernonia (Compositae). Bull. Torrey Bot. Club
97:325-335.

Jones, S.B. 1976. Cytogenetics and affinities of Vernonia (Compositae)
from the Mexican highlands and eastern North America. Evolution
30:455-462.

Kapadia, Z.J. and F.W. Gould. 1964. Biosystematic studies in the
Bouteloua curtipendula complex. III. Pollen size as related to
chromosome numbers. Amer. J. Bot. 51:166-172.

 

Keeley, S.C. and S.B. Jones. 1977. Taxonomic implications of external
pollen morphology to Vernonia (Compositae) in the West Indies.

Keil, D.J., M.L. Luckow, and D.J. Pinkava. 1988. Chromosome studies
in Asteraceae from the United States, Mexico, the West Indies, and
South America. Amer. J. Bot. 75:652-668.

 

Kellogg, E. A. 1987. Apomixis in the Poa secunda complex. Amer. J.
Bot. 74:1431-1437.

 

Kovach, W.L. 1986. M.V.S.P. (A multivariate statistics package),
Version 1.3. Kovach; Indiana University, Bloomington.

Kruckeberg, A.R. 1951. Intraspecific variability in the response of
certain native plant species to serpentine soil. Amer. J. Bot.
38:408-419.

Kruckeberg, A.R. 1967. A hybrid hawkweed from the Olympic Mountains
of Washington. Madrofio 19:126-129.

Kruckeberg, A.R. 1969A. The implications of ecology for plant
systematics. Taxon 18:92—120.

Kruckeberg, A.R. 1969B. Soil diversity and the distribution of plants,
with examples from western North America. Madrofio 20:129-154.

Lewis, W.H. 1977. Pollen exine morphology and its adaptive
significance. Sida 7:95—102.

Martin, P.S. 1969. Pollen analysis and the scanning electron
microscope. SEM/1969: Proc. 2nd Ann. SEM Symp.

Martin, P.S. and C.M. Drew. 1969. Scanning electron photomicrographs
of Southwestern pollen grains. J. Arizona Acad. Sci. 5:147—176.

Martin, P.S. and C.M. Drew. 1970. Additional scanning electron
photomicrographs of Southwestern pollen grains. J. Arizona Acad.
Sci. 6:140—161.

84

Martin, P.S. and B.E. Harrell. 1957. The Pleistocene history of

temperate biotas in Mexico and eastern United States. Ecology
38:468-480.

McVaugh, R. 1943. The vegetation of the granitic flat-rocks of the
southeastern United States. Ecol. Monogr. 13:119-165.

McVaugh, R. 1952. A trip to a botanically little-known area in
Queretaro. Asa Gray Bull. 1:169-174.

Miranda, F. and A.J. Sharp. 1950. Characteristics of the vegetation
in certain temperate regions of eastern Mexico. Ecology 31:313-333.

Payne, W. W. 1978. A glossary of plant hair terminology. Brittonia
30:239-255.

Powell, A.M. and B.L. Turner. 1963. Chromosome numbers in the
Compositae IV. Madrono 17:128-140.

Radford, A.E., W.C. Dickison, J.R. Massey, and C.R. Bell. 1974.
Vascular plant systematics. Harper & Row Publ., New York.

Raven, P.H. 1972. Plant species disjunctions: A summary. Ann.
Missouri Bot. Gard. 59:234-246.

Reader, R.J. 1978. Structural changes in a Hieracium floribundum
population associated with the process of patch formation. Canad.
Jo B011. 56:1-90

 

Robinson, B.L. and J.M. Greenman. 1904. Revision of the Mexican and

Central American species of Hieracium. Proc. Amer. Acad. Arts
40:14-24.

Rollins, R.C. 1979. Dithyrea and a related genus (Cruciferae). Publ.
of the Bussey Institution of Harvard University, pp. 3-32.

Rollins, R.C. and U.C. Banerjee. 1975. Atlas of the trichomes of
Lesquerella (Cruciferae). Publ. of the Bussey Institution of
Harvard University, 48 pgs.

 

Rollins, R.C. and U.C. Banerjee. 1979A. Pollens of the Cruciferae.
Publ. of the Bussey Institution of Harvard University, pp. 33-64.

Rollins, R.C. and U.C. Banerjee. 1979B. Trichrome patterns in Physaria.
Publ. of the Bussey Institution of Harvard University, pp. 64-77.

Rosatti, T.J. 1987. Field and garden studies of Arctostaphylos
uva-ursi (Ericaceae) in North America. Syst. Bot. 12:61-77.

Rzedowski, J. 1978. Vegetaci6n de Mexico. Editorial Limusa S.A.,
Mexico D.F.

85

Sears, P.B. 1917. Amiotic parthenogenesis in Taraxacum vulgare (Lam.)
Schrk. and Taraxacum laevigetum (Willd.) DC. Ohio J. Sci. 62:97-100.

 

Seith, A. 1979. Hair types as taxonomic characters in Solanum, pp.
307-319 in J.G. Hawkes, R.N. Lester & A.D. Skelding, eds., The
biology and taxonomy of the Solanaceae. (Linn. Soc. Symp. Ser.
No.7) Academic Press, New York.

Seith, A. and G.J. Anderson. 1982. Hair morphology and the
relationships of species in Solanium sect. Basarthrum. P1. Syst.
Evol. 139:229-256.

 

Sharp, A.J. 1953. Notes on the flora of Mexico: World distribution
of the woody dicotyledonous families and the origin of the modern
vegetation. J. Ecol. 41:374-380.

Shmida, A. 1985. Why do some Compositae have an inconsistently
deciduous pappus? Ann. Missouri Bot. Gard. 72:184-186.

Skvarla J. and B.L. Turner. 1966. Systematic implications from
electron microscope studies of Compositae pollen-A review. Ann.
Missouri Bot. Gard. 53:220-256.

Snow, A.S. and M.L. Stanton. 1988. Aphids limit fecundity of a weedy
annual Raphanus sativus. Amer. J. Bot. 75:589—593.

 

Solbrig, O.T. 1960. Leaf venation and pubescence in the genus Raoulia
(Compositae). J. Arnold Arbor. 41:259-269.

Solbrig, O.T. 1961. Leaf and nodal anatomy of some Andean Compositae-
Astereae. J. Arnold Arbor. 42:276-294.

Stebbins, G.L., Jr. 1950. Variation and evolution in plants. Columbia
University Press; New York.

Stebbins, G.L., Jr. 1953. A new classification of the tribe
Cichorieae, family Compositae. Madrofio 12:33-64.

Stebbins, G.L., Jr., J.A. Jenkins, and M.S. Walters. 1953. Chromosomes
and phylogeny in the Compositae, Tribe Cichorieae. Univ. Calif.
Publ. Bot. 26:401-430.

Stergios, B.G. 1976. Achene production, dispersal, seed germination,
and seedling estblishment of Hieracium aurantiacum in an abondoned
field community. Canad. J. Bot. 54:1189—1197.

Stone, D.E. 1963. Pollen size in Hickories (Carya). Brittonia
15:208—214.

Takahashi, H. 1986. Pollen morphology of Pyrola and its taxonomic
significance. Bot. Mag. Tokyo 99:137-154.

86

Takahashi, M. 1987. Pollen morphology in the genus Erythronium
(Liliaceae) and its systematic implications. Amer. J. Bot.
74:1254-1262.

Terrell, E.E., W.H. Lewis, H. Robinson, and J.W. Nowicke. 1986.
Phylogenetic implications of diverse seed types, chromosome numbers,
and pollen morphology in Houstonia (Rubiaceae). Amer. J. Bot.

73:103-115.

Turner, B.L. 1967. Chemosystematics: Present and future applications.
Bull. Natl. Inst. Sci. India 34:189-211.

Turner, B.L. 1969. Chemosystematics: Recent developments. Taxon
18:134-151.

Turner, B.L. and R. Alston. 1959. Segregation and recombination of
chemical constituents in a hybrid swarm of Baptisia laevicaulis x
.H. viridis and their taxonomic implications. Amer. J. Bot.
46:678-686.

Turner, B.L. and T.J. Mabry. 1964. Partition chromatography as applied
to taxonomic problems in the Asteraceae. Taxon 13:11-14.

vanWyk, A.E. and I. Dedkind. 1985. The genus Eugenia (Myrtaceae) in
southern Africa: Morphology and taxonomic value of pollen. South
Africa J. Bot. 51:371-378.

Wagner, W.H., Jr. 1968. Hybridization, taxonomy and evolution. In,
V.H. Heywood, ed., Modern Methods in Plant Taxonomy. (Academic
Press, London). '

Webb, A. and S. Carlquist. 1964. Leaf anatomy as an indicator of
Salvia apiana-mellifera introgression. Aliso 5:437—449.

 

Wodenhouse, R.P. 1930. Pollen grains in the identification and
classification of p1ants--V. Haplopappus and other Astereae: the
origin of their furrow configurations. Bull. Torrey Bot. Club

 

Wurdack, J.J. 1986. Atlas of hairs for neotropical Melastomataceae.
Smithsonian Contributions to Botany, no. 63, 80 pp.

APPENDICES

87

APPENDIX I

Citation of specimens used in the compilation of distribution maps.

Hieracium abscissum

Chiapas: Zinacantan, ridge between paraje of Nachih & Zinacantan
Center. 2350 m elev. Breedlove 39793 (M0, TEX).

 

Distrito Federal: N side of Cerro Ajusco. 19° 13'N; 99° 15'W. 3150 m
elev. Beaman 5123 (MSC).

 

Durango: Km 32 on Hiway 40 W of Durango, steep slope E side of road.
Scattered oaks. Warnock 2166 (TEX).

 

Guerrero: Municipio Chichihualco. Asoleadero, ca. 15 km W of Camotla.
Pine-fir forest. 2650 m elev. Feddewa 2836 (TEX).

 

Hidalgo: Cerro de las Ventanas, El Chico National Park. 20° 13'N; 98°
45'W. Fir forest. 2900 m elev. Beaman 5114 (MSC).

 

Jalisco: NW slopes of Nevado de Colima. Spruce-fir zone. 2450 m elev.
McVaugh 10099 (MEXU).

 

Mexico: Valle de Bravo. Pine forest. 2000 m elev. Boege 1751 (MEXU).

Mexico: 3.8 mi W of border between Puebla and Mexico, on free road.
Hillside S face of mts. near the summit of Llano Grande pass of the
Sierra de Rio Frio. Pines. 3150 m elev. Soule 1675 (MSC).

 

Michoacan: Municipio Coalcoman, Barreloso. Cornfield. 2300 m elev.
Hinton 15088 (TEX, MO).

 

Nayarit: 51 mi NW of Tepic on Hiway 15. Wet marsh soil, coastal
plain. Jackson 7199 (TEX).

Nuevo Leon: Crest of Sierra Madre Oriental between Zaragosa & Siberia,
E of Pena Nevada. Oak forest. 2700 m elev. Beaman 6631 (MSC).

Nuevo Leon: Cerro Pena Nevada, ca. 12 km NE of San Antonio Pena Nevada;
30 km E of Dr. Arroyo. Wells 317 (TEX).

88

Nuevo Leon: Sierra La Marta, Galeana. Fir-spruce woods. 3150 m elev.
Hinton 18315 (TEX).

Nuevo Leon: Infiernillo to SW of Galeana near La Cima; Sierra Madre
Oriental. 3000 m elev. No collector listed (MEXU).

Nuevo Leon: Sierra Madre Oriental near Monterrey. Pringle 13853 (TEX).

 

Oaxaca: 14 mi E of Ixtlan. 2900 m elev. Beaman 6498 (MSC).

 

Oaxaca: Mountains, ca. 48 road mi N of Oaxaca and 17 road mi 8 of
pass between Oaxaca and Tuxtepec. 9300 m elev. Cronqgist 9656
(MEXU, TEX).

 

Puebla: 4.8 mi NE of Honey. 20° 15'N; 98° 25' W. Pine forest. 1700 m
elev. Beaman 6497 (MSC).

 

Puebla: 2.2 mi S of the border between Puebla and Mexico, on the free
road to Mexico City. Steep road cut in pine forest. 2800 m elev.
Soule 1668 (MSC).

 

Puebla: 0.4 mi S of the border between Puebla and Mexico, on the
free road to Mexico City. In a state run tree nursery. Pines and
oaks with bunch-grasses. 3000 m elev. Soule 1671 (MSC).

 

Queretaro: 23.2 mi past Jalpan on Hiway 120 to San Juan del Rio. N to
NW facing 80-90° road cuts. Quercus, Pinus, cultivation.
2200 m elev. Soule 1559 (MSC).

 

 

Queretaro: 27.9 mi past Jalpan on Hiway 120 to San Juan del Rio. In
planted pine forest, 30-45° slope facing E. Just at the top of the
pass; Pinus, few Quercus. 2600 m elev. Soule 1561 (MSC).

 

Queretaro: Ca. 0.5 mi N of Amelco on Hiway 120. Series of water tanks
made of soil, banks ungrazed. Soule 1572 (MSC).

San Luis Potosi: Near Puerta de Huerta, Sierra de Alvarez. 22° 08'N;
100° 47' W. Low oak forest. 2300 m elev. Beaman 5113 (MSC).

 

Veracruz: 4.0 mi from route 140 down the road to Misantla in mal pais
(old lava flow). Pinus pseudostrobus. 1450 m elev.
Soule 1638 (MSC).

 

 

Veracruz: 12.1 mi from route 140 down the road to Misantla. Steep,
ungrazed 70-80° E facing cut. Alnus, Quercus, Toxicodendron.
1650 m elev. Soule 1648 (MSC).

 

 

Veracruz: 21.1 mi towards Misantla from Hiway 140 to Xalapa. Plants
growing in cow pasture and along steep road cut. 2100 m elev.
Soule 1651 (MSC).

Veracruz: 13.7 mi towards Puebla on Hiway 140 from Xalapa. Plants
growing under pines in mal pais (old lava flow). Quercus and
Pinus. 2350 m elev. Soule 1654 (MSC).

89

Veracruz: 13.7 mi towards Puebla on Hiway 140 from Xalapa. Plants
growing under pines in mal pais (old lava flow). 2350 m elev.
Soule 1655 (MSC).

 

Veracruz: 19.2 mi towards Puebla on Hiway 140 from Xalapa. Plants
growing under pines. 2350 m elev. Soule 1657 (MSC).

 

Veracruz: 19.2 mi towards Puebla on Hiway 140 from Xalapa. Plants
growing under pines. 1650 m elev. Soule 1658 (MSC).

 

Veracruz: 10 mi S of Orizaba, road to Zongolica. Hill overlooking
Aquila. 18° 41'N; 97° O4'W. 2000 m elev. Turner 15308 (TEX).

 

Hieracium dysonymum

 

Distrito Federal: Extreme SW of the Pedregal de San Angel. 2800 m elev.
Rzedowski 34698 (ASUF).

 

Distrito Federal: Monte Alegre, Serrania del Ajusco, Delegacion de
Tlalpan. 3400 m elev. Rzedowski 37667 (MEXU).

 

Distrito Federal: Capulticla San Bernabe, delegacion Contreras.
"Bosque de oyamel." 2800 m elev. Ventura 2645 (MEXU, TEX).

 

Durango: Km 65 on Route 40 W of Durango. Scattered pines. Warnock
2164 (TEX).

Guerrero: Cerro Teotepec, Municipio Tlacotepec. Pine-fir forest with
grasses. 3200 m elev. Rzedowski 18553 (MICH, TEX).

 

Mexico: At Km 15 between Amecameca and Paso de Cortes. 19° 05' N;
98° 40' W. 3200 m elev. Beaman 5124 (MSC).

 

Mexico: 12 km E of Amecameca, on the road to Tlamacas. "Ladera
andesitica con vegetacion de bosque de Abies." 3100 m elev.
Rzedowski 35593 (MEXU).

 

Mexico: Side of road 0.5 mi past border with Michoacan. Plants in
ungrazed road cuts at the sides of the road. Pine-spruce forest.
2850 m elev. Soule 1586 (MSC).

 

Michoacan: N and S slopes of Mt. Tancitaro. Open pine forest. 10,000
ft elev. Leavenworth & Hoogstraal 1137 (M0).

Michoacan: Road cut on steep slopes above Laguna Larga. San Andres
Mountains, steep 75-90° slope NW facing. Pines. 2875 m elev.
Soule 1583 (MSC).

Michoacan: Road cut next to Laguna Larga. San Andres Mountains. Pines.
2800 m elev. Soule 1582 (MSC).

 

90

Nuevo Leon: Sierra La Marta, on a limestone ridge. 3400m elev. Hinton
18317 (ARIZ, TEX).

Nuevo Leon: San Francisco Canyon, ca. 15 mi SW of Pueblo Galeana.
7500-8000 ft elev. Mueller 415 (MICH).

Nuevo Leon: Ascent to Sierra Infernillo, ca. 15 mi SW of Galeana.
9000-10,000 ft elev. Mueller 826 (MICH).

 

Nuevo Leon: 2.7 mi below the top of Cerro Potosi. In dense numbers
under one pine. 3500 m elev. Soule 1507 (MSC).

 

Nuevo Leon: Road up to the top of Cerro Potosi, ca. 8 mi below the
radio towers at the top. Mesic meadow with pines, few oaks.
2950 m elev. Soule 1501 (MSC).

 

Nuevo Leon: 1.5 mi below the top of Cerro Potosi. 3550 m elev. Soule
1503 (MSC).

Nuevo Leon: Cerro Potosi, 3.5 mi below the summit. Plants in bud, no
flowers. Plants mostly around rocky outcrops where slightly more
mesic. 3425 m elev. Soule 1510 (MSC).

 

Nuevo Leon: Cerro Potosi, 4.9 mi below the summit. SE facing slope
of 30°. Plants mostly in part shade under pines. 3325 m elev.
Soule 1513 (MSC).

 

Nuevo Leon: Mt. Picacho Onofre, ca. 12 km NE of San Antonio Pene
Nevada; 30 mi E of Dr. Arroyo. Wells 401 (TEX).

 

Oaxaca: Ca. 14 mi E of Ixtlan. 2900 m elev. Beaman 6499 (MSC).

Oaxaca: Near Buena Vista, ca. 8 km E of Paso de Cortez. Pinus
hartwegii forest. 3350 m elev. Beaman 6666 (MSC).

 

Oaxaca: Between Ixtlan de Juarez and Llano de los Flores. 2600 m elev.
Beaman 6679 (MSC).

 

Puebla: 1.2 mi W of Texmalquila, just SE of Pico de Orizaba. Pine,
fir, alder, Senecio. 10-11,000 ft elev. Lento 21926 (ASUF).

Puebla: 11.5 mi from Paso de Cortes Monument, down the back of
Popocatepetl and Iztaccihuatl on the dirt rOad. On sides of steep
road cut. Pine-oak. 3300 m elev. Soule 1627 (MSC).

 

Queretaro: 15.5 mi past Jalpan Hiway 120 to San Juan del Rio. N to
NW facing 80-90° road cuts. 1500 m elev. Soule 1558 (MSC).

San Luis Potosi: 55 mi NW of Rio Verde on Hiway 70 to San Luis
Potosi. In full sun next to road, or under pines above road in
ungrazed areas. 2450 m elev. Soule 1550 (MSC).

 

91

Tamaulipas: Sierra de San Carlos, vicinity of San Jose; above La
Vegonia. 3300 ft elev. Bartlet 10055 (MICH).

Tamaulipas: Santa Rita Ranch. 1500 m elev. Runyon 847 (TEX).

 

Tamaulipas: E and S slopes of Pena Nevada. Stanford 2564 (MICH).

 

Veracruz: 7.2 mi from the town of Perote down a side road from the
road up the Cofre. Flat areas under Pinus and Alnus.
3100 m elev. Soule 1636 (MSC).

 

Veracruz: 11.5 mi from the town of Perote on the road up to the
Cofre. N facing 5-10° slope. Pines and bunch-grass.
3550 m elev. Soule 1633 (MSC).

 

Veracruz: Under pines on hillside outside of Villa Aldama. 2650 m
elev. Soule 1660 (MSC).

 

Hieracium mexicanum

 

Distrito Federal: N side of Cerro Ajusco, 19° 13' N; 99° 15' W. 3250 m
elev. Beaman 5121 (MSC).

 

Guerrero: Cerro Teotepec, Tlacotepec. "Bosque abierto de Pinus y Abies
con gramineas." 3200 m elev. Rzedowski 18553 (MEXU).

 

Jalisco: 19 km S of Talpa along the road to La Cuesta. Pine forest.
1400 m elev. Hernandez & Delgado 1331 (MEXU, TEX).

 

 

Mexico: 18 km towards Texcoco, Municipio Tepetlaxatoc. Near the border
with Tlaxcala. 2870 m elev. Garcia 1693 (TEX).

 

Mexico: Mountains N of Huehuetoca, near the border of Hidalgo, ca. 2400
m elev. Gold 21749 (MEXU).

 

Mexico: Ca. 4 mi up the road to the top of Nevado de Toluca, down a
dirt side road. Pine forest with bunch—grasses, 3700 m elev.
Soule 1590 (MSC).

 

Mexico: Ca. 1/2 mi up the road to the top of Nevado de Toluca. Pine
forest with bunch-grasses, 3520 m elev. Soule 1593 (MSC).

 

Mexico: Ca. 1/2 mi up the road to the top of Nevado de Toluca. Pine
forest with bunch-grasses, 3520 m elev. Soule 1594 (MSC).

 

Mexico: Ca. 1/2 mi from Paso de Cortez on dirt road that goes down the
back of Popocatepetl and Iztaccihuatl. Pines and bunch-grass
cover the steep slope. 3600 m elev. Soule 1604 (MSC).

 

Mexico: Ca. 1/2 mi from Paso de Cortez on dirt road that goes down the
back of Popocatepetl and Iztaccihuatl. Pines and bunch-grass
cover the steep slope. 3600 m elev. Soule 1605 (MSC).

 

92

Mexico: 2 mi along the road to Iztaccihuatl trail from the Paso de
Cortez monument. E facing slope. Pines and bunch-grass. 3790 m
elev. Soule 1611 (MSC).

 

Mexico: 2.7 mi along the road to Iztaccihuatl trail from the Paso de
Cortez monument. E facing slope. Pines and bunch-grass. 3800 m
elev. Soule 1612 (MSC).

 

Mexico: End of road at the base of the trail up Iztaccihuatl. Above
timberline. N facing slope. Bunch-grasses cover the slope.
4000 m elev. Soule 1614 (MSC).

 

Mexico: 3.8 mi W of border between Puebla and Mexico, on free road.
Hillside, S face of mts. near the summit of Llano Grande pass of
the Sierra de Rio Frio. Pines. 3150 m elev. Soule 1676 (MSC).

 

Mexico: 5.3 mi W of border between Puebla and Mexico, on free road.
Hillside, 8 face of mts. at the summit of Llano Grande pass of the
Sierra de Rio Frio. Pinus, Qgercus. 3275 m elev.

Soule 1678 (MSC).

 

 

Mexico: 5.3 mi W of border between Puebla and Mexico, on free road.
Hillside S face of mts. at the summit of Llano Grande pass of the

Sierra de Rio Frio. Pinus, Quercus. 3275 m elev.
Soule 1679 (MSC).

 

 

Mexico: Nevado de Toluca, just below timberline, below the dirt road to
the top. Pines and bunch-grasses. 4000 m elev. Soule 1681 (MSC).

 

Michoacan: Summit of Cerro San Andres, ca. 12 km (straight line
distance) N of Ciudad Hidalgo. 3589 m elev. Beaman 4287 (MSC).

 

Puebla: Pico de Orizaba, SW side of the mountain. 3860 m elev.
Beaman 2516 (MSC).

 

Puebla: Lado Norte de Pinal, entre rocas con Pinus oaxacana y
H, teocote. 2800 m elev. Boege 2137 (MEXU).

 

 

Puebla: Pico de Orizaba. 11-12,000 ft elev. Priggle 8550 (MEXU, M0,
MSC, TEX).

 

Puebla: 9.9 mi from Paso de Cortez monument, down the back of
Popocatepetl and Iztaccihuatl, dirt road. Pine forest, few bunch-
grasses. 3450 m elev. Soule 1620 (MSC).

Veracruz: 6.3 mi from the town of Perote on the road up Cofre de
Perote. Planted pines. 3050 m elev. Soule 1629 (MSC).

 

Veracruz: 6.3 mi from the town of Perote on the road up Cofre de
Perote. Planted pines. 3050m elev. Soule 1630 (MSC).

Veracruz: 11.5 mi from the town of Perote on the road up to the
Cofre. N facing 5-10° slope. Pines and bunch-grass. 3550 m elev.
Soule 1632 (MSC).

 

93

Hieracium pringlei

 

Agua Caliente: Sierra del Laurel near the border to Jalisco, ca. 10 mi.
SE of Calrillo. Oak forests near summit. 2600 m elev. McVaugh
18450 (MICH).

Chiapas: Clinica Yerba Buena, 2 km NW of Pueblo Nuevo Solistahacan.
Chromosome voucher. Pine-oak-sweetgum. 5400 ft. elev. Breedlove
and Raven 19921 (MICH).

Chiapas: Near Piedracitos, ca. 7 km E of San Cristobal las Casas. Oak-
pine forest. 7200 ft. elev. Breedlove 8006 (MICH).

 

Chihuahua: Basaseachic Falls area. 6400 ft. elev. Pinkava 13265
(ASUF).

Chihuahua: La Bufa above Cusihuiriacha. Pringle 1585 (MEXU, MICH, MO,
MSC).

 

Coahuila: Sierra Santa Fe del Pino, 28° 12' N; 103° 03' W. Juniper-oak
forest. 2100 m elev. Johnston 11255 (TEX).

Distrito Federal: Base of Sierra de Ajusco. 2400 m elev. Pringle 7211
(M0, MSC, TEX).

Distrito Federal: N of Cerro de Santa Catarina, around Santa Marta
Astahuacan. 2500 m elev. Rzedowski 30821 (MSC).

Durango: 32.7 mi W of Durango on creek terraces. Pine—oak Forest.
Johnston 2693 (TEX). '

 

Guanajuato: 8 km NE of Santa Rosa. 2400 m elev. McVaugh 24113 (MICH).

Jalisco: Nevado de Colima. 3070 m elev. Eiten & Gregory 289 (MICH).

Jalisco: Cerro de Talcozagua, on Sierra de Tapalpa, 2-3 km E of
Tapalpa. 19° 57'N; 103° 45'W. 2000 m elev. Iltis 25 El; 754
(MICH).

Jalisco: Sierra de Cuale, SW of Talya de Allende. Pine forest. 2200 m
elev. McVaugh 14262 (MICH).

Jalisco: Las Vidrieas, 10 km NW of El Platanar, Municipio San Martin de
Bolanos. 2450 m elev. Rzedowski 26181 (MICH).

Jalisco: Rincon de Manantlan. 19° 33' 30" N; 104° 13' 15" W. Cloud
forest. 2250 m elev. Sorensen 7884 (MICH).

 

Jalisco: Volcan Colli, Sierra de la Venta, 10 km W of Guadalajara.
Villareal 4649 (MICH)~

 

Jalisco: Mountains E of Mamantlan, ca. 15 mi SSE of Antlan via Chante.
8300 ft elev. Wilbur 1803 (MICH).

94

Michoacan: Vicinity of Morelia. 2100 m elev. Arsene 6626 (M0).

 

Michoacan: Logging road 24 miles past border with Michoacan. E facing
slope. Pine-oak forest. 2440 m elev. Soule 1575 (MSC).

 

Michoacan: Logging road 24 miles past border with Michoacan. E facing
slope. Pine-oak forest. 2440 m elev. Soule 1576 (MSC).

 

Michoacan: Ca. 10.5 mi past the border with Mexico. 70° E facing slope
very wet, lots of moss, liverworts. 0n cut and in pine-oak forest.
2390 m elev. Soule 1577 (MSC).

 

Michoacan: Ca. 10.5 mi past the border with Mexico. 70° E facing slope,
dry, sandy soil. Pine-oak forest. 2390 m elev. Soule 1578 (MSC).

 

Michoacan: 4.8 mi down road to Los Azufres off Hiway 15. Slopes of
0-45° facing all directions, ungrazed. Pine—oak forest. 2425 m
elev. Soule 1579 (MSC).

 

Nuevo Leon: Ca. 13 km SSE of Galeana Santa Rita. 1980 m elev. Hinton
18300 (TEX).

Nuevo Leon: Galeana, Sierra La Marta. 3400 m elev. Hinton 18317
(ASUF).

 

Nuevo Leon: Dulces Nombres, just E of border with Tamaulipas. Oak
forest. 1950 m elev. Meyer 2620 (M0).

 

Nuevo Leon: On Highway 51 (road to Dr. Arroyo) 5.6 mi S of junction of
Hw 51 & Hw 58. Dry canyon with pinyon pines. 6400 ft. elev.
Smith 2103 (TEX). ‘

Nuevo Leon: 68 mi N of Zaragosa on gypsum soil with pockets of high OM
under pinyon pines. 10° NE facing slope. 2000 m elev. Soule 1516
(MSC).

 

Nuevo Leon: 68 mi N of Zaragosa on gypsum soil with pockets of high OM
under other plants. 30° NE facing slope. Pines. 2000 m elev.
Soule 1519 (MSC).

 

Nuevo Leon: 68 mi N of Zaragosa on gypsum soil with pockets of high OM
under other plants. 30° NE facing slope. Pines. 2000 m elev.
Soule 1520 (MSC).

Nuevo Leon: 65 mi N of Zaragosa on gypsum soil with pockets of high OM
under pinyon pines. E facing 5-10° slope. Pinyon pine, juniper.
2125 m elev. Soule 1523 (MSC).

Nuevo Leon: 61 mi N of Zaragosa. Gypsum soil with pockets of high OM
under pinyon pines. 5-35° slope facing NW to W to SW. Above road
only. Pinyon pine, juniper, manzanita. 2225 m elev. Soule 1524
(MSC).

 

95

Puebla: Cerro de Gentile. 7000-8000 ft elev. Purpus 3848 (M0).

 

Puebla: On La Derrumbada (landslide hill), uncultivated hill on the
west side of the road to Guadalupe Victoria off Hiway 140.

Municipio de Guadalupe Victoria. Pines, oaks, and agaves. 2600 m
elev. Soule 1665 (MSC).

 

Puebla: On La Derrumbada (landslide hill), uncultivated hill on the
west side of the road to Guadalupe Victoria off Hiway 140.

Municipio de Guadalupe Victoria. Pines, oaks, and agave. 2600 m
elev. Soule 1666 (MSC).

 

Queretaro: Cerro Zamorano, 1 km SW of Cumbre. Spruce forest. 3100 m
elev. McVaugh 402 (MICH).

 

San Luis Potosi: Near San Luis Potosi. 7000 ft. elev. Parry H Palmer
552 (MO).

Tamaulipas: Pico del Diablo, vicinity of Marmolejo. Bartlett 10909
(MICH).

 

Veracruz: 4.0 mi from route 140 down the road to Misantla in mal pais
(old lava flow). Pinus pseudostrobus. 1450 m elev. Soule
1639 (MSC).

 

Veracruz: Under pines on hillside outside of Villa Aldama. 2650 m
elev. Soule 1659 (MSC).

 

Veracruz: Under pines on hillside outside of Villa Aldama. 2650 m
elev. Soule 1660 (MSC).

 

Zacatecas: Ca. 20 km W towards Tlaltenango from road junction S of
Julpa. Oak-pine forest. 2400 m elev. McVaugh 25631 (MICH).

 

Hieracium schultzii

 

Chihuahua: Hills near Chihuahua. Pringle 771 (MSC).

 

Coahuila: 17 km N of Rio El Jardin, 29° 10' N; 102° 46'W. Pinyon
pines. Johnson 11900 (TEX).

 

Distrito Federal: Desierto de los Leones. Lyonnet 407 (MO).

Durango: City of Durango and vicinity. Palmer 653 (MO).

 

Durango: San Juan de Michis, a1 cerro "chihuahullia." Chavez HQ (TEX).

Hidalgo: Tepeapulco. 2550 m elev. Ventura 1515 (ARIZ, ASUF, XAL).

 

Mexico: 13.3 mi down the road to Valle de Bravo, road that is closer to
Toluca, but not from Toluca. W facing cut. 2810 m elev. Soule
1588 (MSC).

96

Mexico: 6 km W of San Cristobal Ecatepec, parte alta de la Sierra de
Guadalupe. Vegetacion de matorral. 2600 m elev. Rzedowski 32166
(MEXU, MSC, XAL).

Mexico: 8 km E of Coatlinchan. 2250 m elev. Rzedowski 35148 (MEXU,
MSC).

 

Mexico: Parte SE del Cerro del Pino. 2600 m elev. Rzedowski 34801
(MEXU, MSC, XAL).

 

Mexico: Valley of Mexico, Pedregal. Bourgeau gag. (MSC).

Oaxaca: S slope of E1 Pasajuego, ca. 25 km NE of Oaxaca on Hiway 175.
Oak-pine forest. 2000-3000 m elev. Conrall 3085 (MEXU).

 

Oaxaca: Sierra de San Felipe. 10,000 ft elev. Pringle 4715 (MEXU).

 

Pueblo: Hills near Amozoc. 9000 ft elev. Pringle 9360 (M0).

 

Pueblo: San Luis Tultitlanapa, near Oaxaca. Purpus 3131 (M0).

 

Queretaro: Ca. 0.5 mi N of Amelco on Hiway 120. Series of water tanks
made of soil, banks ungrazed. Soule 1572 (MSC).

 

San Luis Potosi: 55 mi NW of Rio Verde on Hiway 70 to San Luis Potosi.
Under pines above road, or in full sun next to road, in ungrazed
area. 2450 m elev. Soule 1551 (MSC).

 

San Luis Potosi: Cheifly in the region of San Luis Potosi, 22° N.
6000-8000 ft elev. Parry 553 (MO). '

San Luis Potosi: Microwave hill about 15 mi SE of San Luis Potosi. In
road cut at base of hill. N facing slope to 70°. 2500 m elev.
Soule 1549 (MSC).

 

Veracruz: 14 km NE of Huayacocotla, 20° 36' N; 98° 28' W. 1650 m elev.
Vasques HQ (XAL).

Veracruz: Under pines on hillside outside of Villa Aldama. 2650 m
elev. Soule 1661 (MSC).

Zacatecas: 16 mi E of Concepcion del Oro, on upper N side of Sierra del
Astillero, 24° 38' N; 101° 08' W. 8200 ft elev. Hendrickson 13342

 

(TEX).

97

APPENDIX II

Raw data used in statistical analysis of the vestiture.

The presence or absence of each type of vestiture was established
after all SEM work was completed and the resulting micrographs surveyed.
The results were scored and entered in a data matrix for statistical
analysis using MVSP (see text). This matrix is presented below.

Scoring evaluated the presence of a character as 0.99, absence of
the character as 0.01. Lengths of glandular hairs were measured and
their average entered. Barb lengths were evaluated and long barbs (see
text) were scored at 0.99, short barbs at 0.01. Rows consisted of each
sample and its characters, columns were determined by the following
categories:
c0 species combined with collection number to define each row, limited

to eight characters, printed out but not recognized for analysis by

the program,
c1 phyllary-stellate hairs,
c2 phyllary-—glandular hairs,
c3 phyllary--barbed hairs,
c4 phyllary--waxes,
c5 stem below the head--g1andular hairs,
c6 stem below the head--stellate hairs,
c7 stem below the head--barbed hairs,

c8 stem below the head--waxes,

c9

c10
c11
c12
c13
c14
c15
C16
c17
c18
c19
c20
C21
c22
c23
c24
c25
c26

c27

upper

upper

upper

upper

upper

lower

lower

lower

lower

lower

surface
surface
surface
surface
surface
surface
surface
surface
surface

surface

of
of
of
of
of
of
of
of
of

of

stem at

stem at

stem at

stem at

stem at

the base

the base

the base

the base

the base

the
the
the
the
the
the
the
the
the
the
of

of

of

of

of

pappus color white,

pappus color tawny,

leaves basal only,

98

1eaves--glandular hairs,
leaf--stellate hairs,
leaf--barbed hairs,
leaf--barb length,
leaf--waxes,
leaf--glandular hairs,
leaf--stellate hairs,
leaf--barbed hairs,
leaf--barb length,
leaf--waxes,

the plant--glandular hairs,
the plant-stellate hairs,
the plant--barbed hairs,
the plant—-barb length,

the p1ant--waxes,

leaves basal and cauline.

 

 

Data matrix used for MVSP statistical analysis (see text).

99

9

N 0 00HHH 000000000‘

0. O O 0. O. O
OOOOOOOOOOOOOOOOOOO
I-QI-iu-‘v-i 0—4—1—a-n—u—ao-4—q—uo
000000 00000000000
OOOOOOOOOOOOOOOOOO

000 0 0 0 000

dddddddddooooooooo

c26 c

c25

O‘F‘H F“"-fi—"—4—4~u-4

o—do-tv—uv-ta—cO‘Ch
000000000000000000
OOOOOOOOOOOOOCOOOO
ONO—{Ho-iv-du—q—d—ov—a—aq—no O‘HI-fl
00000000000000 000
OOOOOOOOOOOOOOOOOO
u—c—d—O—flq—dv-d—nv-Qgc‘ 00‘0““
00000000 000000000
000000000000000000
0

8.3 aaa aaoaeaoooss
OOOOOOOOOOOOOJm—zmm

v-flv-QHH O‘u—dv-fiv—c

.4.-o -4
000000 00000000800
O .0. 0.0 D... O
OOOOOOOOOOOOOOOOOO
u—cv-tOsCh H000—1—1v—1—u—c—1—‘a—t—uox

000000000000000000:
000000000000000000

c20 c21 c22 c23 c24

c19

00—0—‘8H—0—4 00—‘r-t0000v-4
00000 000 000000000
0000000000000000000

9 0.01 0.01 0.99 0.01 0.99 0.01 0.99 0.01

a—cu—aH—o—t v—v—ac—n 0‘0 a—t—no—‘o—cu-co—to—c

00000000000 OOOOOOO

OOOOOOOOOOOOOOOOO
m In

NNNNNNN 088838882

.0 O O 0.. O 0...... O

HHv—c—qv-qu-o—co—oo-qN—a—n—co-q—tNO

c17 c18

O
0

c16

0M —‘
00000000000000000
00000000000000000

c15

c8
absl408 0.99 0.50 0.01 0.99 0.99 0.99 0.01 0.99 0.01 0.01 0.80 0.01 0.99 0.01 0.01 0.90
9
9
0
0
O
O
0
O
99

Id Hp‘fifl-‘Hfi
owooooooooooooooo
ooooooooooooooooo

a mo oooo
goasgoosaooowo

c14

cl3

00000000000000

c—tv-iv-dr-t OOH—{HfiHH—fi

d
0
0
H
000003000000000
00000000000000000
0
m
H
OH
0

c12

00° sea 33888

0:00:
—:OOOo—o—4H—o p:p-:NNO

cll

1.3)

O‘fi—cfi—d—Ir-I

aooooooc
oocoooooooo

v—IHH

00000
00000

c—to—do—Qo—Oc‘

00000
00000

clO

Fin-cl“

00000800000
00000000000

c9
9 0.01 0.01 1.20 0.01 0.99 0.01 0.01 1.50
O
O

.99 0.01 0.01

0 0
. 00300 03030
000000000000000000
wa—au-a
000000800 8880 8000
0000000000—‘0-H-‘00000

O‘ 0 0000‘
000000000000000000

d2 r-m—a
088888233 RmRSSMNo
00000000000000000
0‘ 0‘ 000000“
000000
0‘ a~ '-
0:8:c~E;c>

00000

c4 c5 c6 c7

0
0
0.
0.
O
9 0.
O
0
0
0.
0

c2 c3

c1

pri1664 0.99

ah31573 0.99 0.30 0.01 0.99 0.3
O
O
O.
0
0

dysl412 0.99 0.30 0.01
dysl421 0.99 0.57 0.01
9
99
99
99
nu11519 0.99 0.73 0.01
nu11523 0.99 0.58 0.01

abslb48 0.99 0.20 0.01
sch1409 0.9

pr3U521 0.99

mex1614
pr11575