COHTROL OF FUSAREUM WHI
OF CARHATICNS BY CHEMOTHERAPY

Thai: for tho Degree of M. S.
M2CHIGAN STATE COLLEGE

David Aloysius Besiey
1952

 

This is to certify that the

thesis entitled

CONTROL OF FUSARIUM WILT OF
CARNATION BY CHEMOTHERAPY

presented by

David A. Bosley

has been accepted tofiards fulfillment
of the requirements for

M. .3. degree 1n_Elan.t_P_athology

 

 

CCNTROL C? FUSERTUM WILT OF CARVATICNS FY CWEWOTV?RAPY

By

DAVTD ALCYSITS TOQLEY

A THESIS

Submitted to the School of Graduate Studies of
Michigan State College of Aericulture and
Apolied Science in partial fulfillment
of the requirements for the degree

of
MASTER OF SCIENCE
Department of Botany and Plant Pathology

1952

THE-sits

(‘1’ T [A1, V“ ”V7,“? T n
Auithv.rr1h:)xlo ‘._ MT.)

The author wishes to excress his apnreciation and gratitude
to Dr. John R. Vaughn for his helpful suggestions and guidance
throughout this study. The author is also indebted to Dr. Donald
J. Dezeeuw for his critical evaluation of the data presented in
this thesis.

Other members of the Department of Botany and Plant Pathology
to whom the author wishes to exnress his thanks for helo in various
phases of his work, are Dr. George Stein auer for his aid in
nutritional problems, and William Flomnarens for invaluable

assistance in serial dilution studies.

0

Dr.’”alter J. Haney, of the Denartment f Horticulture, helped
with information concerning commercial carnation propagation techniques.
Snecial consideration and gratitude is due Yoder ‘ros., Inc.,
“aroerton, Ohio, who kindly ofpered 1,100 unrooted carnation cuttings

without which this study would not have been made.
Carbide & Carbon Chemicals Elvision, of Union carbide % Carbon
Corn., New York, N. Y., kindly sunnlied the author with sufficient

quantities of Exnerimental Chemotheraneutant 11“? F to carry out

this experiment.

:' 'n ‘
23.31;?) :‘l

AN ABSTFACT OF A TH

ES 5 ETTITLED " CONTROL OF PUSk.-IUM
MILT OF CARNATI

ON EY CHEL‘ZO'IHERAPY. "

A ohemotheraoeutant, h— chloro-B, S—di methyl phenoxy ethanol (Exnerimental

'Cherot heranautant 1182l ), was applied in soil solution to four varieties of

Carnations, Neptune, Jupiter, Hides, and Vulcan, in an effort to control

C‘)

Fusarium wilt. The concentrations of E. C. 11 2* were 1:16,875; l:33,750;
and 1:67,EOO.

Visual symptoms of the disease caused by Pusarium dia nthi , plus an

 

adaptation of th e commercial method of "cultu iring" cutt int“ s were used to
arrive at total percent disease in treated and check plots. Connercial
culturing of cuttings consists of selection of shoots or cuttings from
apparently healthy plants, plat‘ hasal stem sections from these
on tings on to potato dextrose agar, incubation at temperatures favorable

'l

to the growth of Fusarium ani identification of the disease on the Petri

(L

ish.

’D
(.0

The total percent disease in 288 plants which received E. C. 1182?
until the time of inoculation was 31.97. .or 288 plants which continued
to receive applications of E. C. 1182?, the total Percent disease was 8.1%.

4 1

TWO vaTiC' :6 S, Vulcan ani NOT‘tun

(o

, failed to become infected after i_noculaticn

'l V

in the continuous—treatment group. In tne Check plants, the total percent
Cut flower records of flowers out per plant, ca e.do n he CcT v.einht
grading system as acnlied to oarnations, indicctcs that hea.lt hy plan
out-produce disea as ed rlants by a noticeable margin. The data shows that
2.72 flowers per plant were out :from healthy check plants, while only 1.22

were cut from infected check rlants. The disease apparently lessened the

number of flowering shoots produced by the plant.

‘

Indications are tnat the afore- mentioned material will effectively
bring about a temporary resistance in the carnation to the entrance of
Tusarium wilt when it is used in con: ination with disaase-free cuttings

and strict sanitary measures.

Approved ‘y a?. y{;L~AHn/€:--

Wee-g.” 01,

WWW

I“m“CDUCT ON

The principle of chemotherapy for the control of plant
diseases s, while not a new cor cent, has received wide ouolicity
and interest during the past ten years. 'With particular
reference to vase ular diseases which are masked under certain
environmental condif ions, treatment by means of chemotherapeutants
shows great prose in alleviating serious diseases of this type.

Dimond eta ,(9) state that chemOAtn eraneutic control migh
he a result of direct fungotoxicity, inactivation of toxins
produced by the pathogen increase in the host's resistance to the
disease by altering protoplasmic factors or unsetting nu ritional
balance, or by a change within the host of the chemotheraneutant
to a configuration more toxic to the pathogen.

“ecause of the poss1bi li ti es of commercial annlicaticn of the
theory 0. c hemctherany to greenhouse plants, as shown by Stoddard
and Uimond (20), this study was started to determine the degree of
control of a disease of carnations by means of so 1 annlioat ions
of a chemotherapeutant. Also, this study att nmt to distinguish
between the tynes of control which this material mi: t exhibit.
Cut flower records are included to dem onstrate differences in
production between healthy and diseased nlants.

While methods for determining the presence of:ystemic

fungicides in the host are not as ye+ defined or standardized,

3.
the decree of control of the disease at aif erent concentrations
of the chemotherapeutant as comparei with untr ated, inoculated
plants is indicative of therapeutic activity, even though the

ex ct mode of action of the material is unknown.

L'.

HISTORY OF CTEFCTWTRAPY
Chemotherapy of Local Tiseases

In 1891, Eclley (3) showed that potato scab, a tumor on the
tuber, could be prevented from appearing by soaking the diseased
potato in a mercuric chloride solution. Fifty years later,

Strong and Cation (21) cured rust galls on cedar with sodium
dinitro-o-cresylate, and chemotherapy of local or tepical diseases
began with renewed interest.

Positive action on bacterial crown gall by tOpical applications
of sodium dinitro—o-cresylate has been reported by Ark (l); with
penicillin by ?rown (S) and with penicillin and streptomycin by
Hampton (ll).

According to Yarwood (25), in 1931, Hamilton cured apple scab
with lime sulfur and organic mercury early in the disease year, and
his findings went into commercial adoption. Newton et al., (16) found
that catechol injected into wheat leaves, four days after inoculation
with urediosnores reduced rust develonment by almost fifty percent.
Yarwood (25) found that five-day old uredial pustules of been rust,
snapdragon rust, and sunflower rust were killed, without host injury,
by exposure of the plants to the vapors from lime sulfur solutions or
from dilute hydrogen sulfide gas.

According to Yarwood (2S), Hiltner was the first to recognize

that deep infections in seeds could be cured by application through

the uninjured seed coat.

‘JI

He cured rye seeds, infected with FUSarium, by soaking them in
mercuric chloride.

Organic mercurials soon replaced mercuric chloride, and
thousands of bushels of cereal seeds, infected with F‘sariim,
delminthosnorium and Ustilago, are treated in many parts of the
world.

In l9h0, Nugent and Cook (17) cured rape seeds infected

7

with bacterial black rot oy fumigating the seeds with chlorooicrin.

Systemic Chemotherapy on Local Diseases

The application of a fungicide to a given leaf, stem, or root
surface ani the translccation of that material to the site of
infection is as close to a definition of a systemic fungicide as
probably any other at the present time. Evidence of chemotherapeutic
activity on local or tonical diseases was noted by Kinney (lb) who
demonstrated that ootato tubers were more free of scab on plants
sprayed with “ordeaux mixture than tubers on unsnrayed plants,
probably because tubers on snrayed vines were higher in cooper
content than those on unsprayed vines.

Me New and Sundholm (15) immersed one leaf on each tomato
plant in suspensions 0? several 1 (substituted phenyl) - 3,
S—dimethyl - h—nitronyrazoles for thirty-six hours. The treated
leaf was severed from the plants and the other leaves were
inoculated with Alternaria solani. Appreciable reduction of
disease on the untreated leaves showed clearly that the compound
had been absorbed through one leaf and translocated to the other

leaves.

Systemic Chemotherapy on Systemic Diseases

A systemic disease li a a systemic chemotherapeutant is one

D

(

in which the pathogen itself or it. toxic by—products move through
the conducting system of the plant away from the infection court.

Howard and Caroselli nrovei that bleeding canker of hardwoods
is a toxin-induced disease which could be mitigated if not cured
by antidoting the toxin with injections of diaminoazobenzene
.ihydrochlorid .

Pusarium wilt of tomato and carnation have also received
attention by Dimond et al., (9), Stoddard and Dimond (20). The
following chemotherapeutants have shown good to excellent control
of one or the other of these diseases: h—chloro-B, S-dimethyl
phenoxy ethanol, 2—norcamphane methanol, 8—quinolinol sulfate,
n-ootadecyl trimethyl ammonium pentachloroohenate and N-(h—nitrOphenyl)
-3, h—dichloro benzene sulfonamide. Toxity to spores hears little
or no relation to chemotherapeutic activity against Fusarium wilt

of tomato. The carnation appears to activate h-chloro—3, S-dimethyl

phenoxy ethanol for Fusarium wilt (13).

7.

. MATERIALS dUD YETHCDS

A chemotherapeutant, h—chloro—3, S—dimethyl ohenoxy ethanol,
previously shown to possess active control over Fusarium wilt of
carnation (9), (13), and (20), was used in comparison to untreated

plants in the course of this study.
Phytoxicity Trials

From July 7, 1951 until August 29, 1951, a period of
approximately seven weeks, five different concentrations of Experi—
mental Chenotherapeutant 1182 F, as the above material is known,
commercially, were applied in solution to carnation plants well
established in h-inch clay pots in the greenhouse. The concentrations
were as follows: 1 part E. C. 1182? to 16,875 parts of water;
1:33,?50; 1:67,§OO; 1:13S,OOO; and 1:270,000. Four varieties of
carnations were used: Ida, Miller's Yellow, Northland, and an

unnamed white-flowered variety.

9

1"
D

The above concentrations of 1. C. 1182? were applied.in water
solution with a watering can. Approximately 50 ml. of eacn
concentration were put on each h—inch pot twice a week for a
period of four weeks. At the end of this time, double the
quantity of solution or 100 ml. Jere applied.

On August 28, at the end of the seven weeks treatment, there

was no indication of phytotoxicity, loss of vigor, or disease

symptoms in any of the plants. There were three replications
ith four check plants in each treatment.

Preparations were made to receive a shipment of 1,100
"cultured" carnation cuttings, generously donated by Yoder Bros.,
Inc., Varberton, Ohio. ,The procedure was the steam sterilization
of concrete propagation benches, sand, 3-inch clay pots, acid
peat, concrete greenhouse benches and tools. All were steam
sterilized separately for five hours at approximately 2000F., with
high pressure steam.

Upon receipt of the unrooted carnation cuttings from Yoder Vros.,
Inc. on August 21, they were immediately placed in the prepagating
bench containing fine washed sand. Fifty random basal stem tissue
sections from 1,100 cuttings were plated on potato dextrose agar
containing two percent dextrose and incubated at 30C 0., for seven
days before readings were made. None of the plates showed any
growth of Fusarium spp.

0f the total of 1,100 cuttings, there were 275 cuttings each
of four varieties ------ Jupiter, Midas, Neptune, and Vulcan.
Treatments were begun immediately on 650 uprooted cuttings while
hSO cuttings were maintained as checks. The concentrations of
E. C. 1182 F used were 1:16,87F; 1:33,7SO; and l:67,§00. These
solutions were first watered on the unrooted cuttings August 21,
and were applied twice weeily after that for a period of forty-five
days. When the cuttings had formed roots approximately one—half
inch long in the prOpagating bench, a liquid fertilizer, Plant
Marvel, with an analysis of 12-31-13 was watered on the cuttings

twice a week for a period of approximately three weeks.

Cuttings were left in the propagation oer ch and fed liquid
fertilizer so that sufficient growth of roots and too would take
place enabling a top "pinch"o of the terminal shoot wh ch would
provide an additional infection court wound to supplement the
customary root injury type of infection court when the uttings
were inoculated.

Five isolates identified as those of Vusarium dianthi by

 

Dr. Ames, of Massachusetts State Coll. were transferred and
maintained on potato dextrose agir minus Sigar Ccto r20.

in order to build up a sufficient quanti _ty of inoculum for the
large number of cuttings involve., it was decided to use
convenient proced re ani ed W11 ment for grow:ing f ingus inoc11um.
Dotato ju ce we. made by boiling 500 grams of cleaned potatoes
with skins intact in one liter of distilled wa+ er and then
straining the resultant medium through cheese cloth. This
solution was then placed in a h-liter oyrex syrum bottle. The
rubl er stopper in the bottle contained holes fw the replacement

glass tubing which connected to a vacuum pump. Another tube
filled with cotton provided for the intake of one liter of air
per minute per l_iter of melium which would be forced through the
solution. The entire apparatus was autoclaved at fifteen pounds
of pressure and at QhCC F. for thirty minutes. Voaming was
prevented by the addition of a small quantity of lard oil oefore
autoclaving. ”he apparatus used +o ma {e large quanti ies of
inoculum is shown in Vigure 1.

When the sterile medium had cooled sufficiently, sections of

fusariu dianthi 5mm. in diameter, were transferred from netri

 

 

Pig. 1. Culture apparatus for growing fingus

inocu'un.

10
dishes to the medium in the jar. The vacuum pump was started and
the liquid medium was constantly aerated for a period of three days.

Purina the course of the three-day period, loops of the medium were

(1)

removed every six h urs t- determine the exact time when the

H)

macrosoores o Vusarium dianthi had begun to germinate. The

 

presence of germ tubes suhstantiatei the viability of the culture

‘I

and also indicated the pest possible time for inoculation of the

rooted carnation cuttings.
Tethod of Inoculation

The method of inoculating the rooted cuttings was as follows:
Fefore inoculation, the potting bench’used in the greenhouse was
washed thoroughly with a 1—500 solution of formaldehyde. When
microscopic examination of the medium showed an abundance of visible,
germinating macroconidia, 500ml. of this inoculum.was diluted to a
liter of incculum with unsterile distilled water and then taken
to the greenhouse for inoculation. The carnation cuttings, which
by now had a heavy root growth and approximately seven to eight
inches of top growth, were removed fairly roughly from the

prooagatin

UQ

bench so that there would be ample root injury thus
allowing entry of the mac-oconidia germ tubes into the plants.
Cuttings were then too "pinched" thus providing an aiiitional
infection court for the disease which, according to literature,
enters the carnation plant almost entirely through wounded tissue.

The entire cutting was immersed in a liter of snore suspension
for approximately thirty seconds. Check plants were dipped for

thirty seconds in distilled water. After being reroved from the

ll.

inoculum, the cuttings were rlaced in 3-inch rots. The soil
mixture consisted of two parts peat, three parts clay loan, and
one rart sand, all 0” which had been steam sterilized serarately.

Cupernhosnhate, at the rate of one h-inch notful nor who ltarrel

I
\

L-J.

of soil mixture, was included before rcttinc. After nott n3, the

tear sterilised neat un to the too 0? the cots.

nlants were buriei in

0)

FY

L1.

in which the inoculated

‘ + 4-,,» J. ° t1 r. “In A.
I‘v‘ i:. hav- ."I . P 71 _. my 3 (N11 ‘3-
;.'.J $1-)“ \_‘4-O-U.‘reu :1;— .L~J 3’1 ‘ -r‘.L~'. At";

nlarts were nlaced, were raintained at 600 V., with awhient day
temperatures prevalent durine the late fall and early shrirg.
Check slants were inoculated by the cane method as doccrited

acove. Uninoculated, but chemotheraneutically treated nlsnts

were irrw

Losses During nronagtation

Tron a total of 6?0 unrooted treated cuttings originally
placed in the nroracating bench, eighteen cuttings failed to root

for unknown causes. A severe infection of Alternaria dianthi

 

develoned in the nronagatin: Perch, and as a result, a total of

V

fifty—six cuttings were lost to t-is disea

U)

e. Since the annlication
of any fungicides to control A. dianthi would nullity the results

of th

s owneriment, it was decided to write of? the losses. Thus

H.

the final numher of rooted cuttings inoculated in the treated nlots
was F76.

Losses in the inoculated checTr group and in the absolute checks
amounted to forty—seven plants. The total nuober o” rooted cuttinds

potted in the former group was 18h, and in the latter, 219.

To discover if E. C. 1182 F had any direct fungotoxicity
or merely controlled Fusarium wilt by immunizing the host or
makinc it resistant to the disease, it was decided to senarate

the treated groun of L".76 nlants into two equal plots as follows:

J

(f

293 plan 5, all of which had received Ji-weekly annlications of

E. C. 1182 3 for anoreximately forty-tive days, continued to
receive these annlications at three different concentrations—-—-——
875; 1:93,7F0; andl:67,§00, for a neriod of 125 days or
from their initial annlication ir the pronacatirg bench, August 21,
until t‘e final annlication on January 24, 1952.

The remaining qroun o? 288 treated—inoculated plants was
senarated from the continous-treatment plot, ani these nlants
received no treatnent of any kind after inoculation.

Several liters of inoculum grown in the same manner as
described nreviously, and from the same original transfer of
cultures received from Fassachusetts, were used to re-inoculate
the rlants on December 5, one month after the date of the initial
inoculation.

At this time, each FOOml. o? notato juice containing a snore
susnension of macroconidia was diluted with the same quantity of

unsterile distilled water and annroxiwately ten ml. of this

inoculun was noured into a hole in the soil 0? each rot near the

"J

stem

0

t the plant. fiho holes were waie by ianminq $0t 135913

R

into the soil so that there would be some root injury, thus

providing an inf-ction court favorable to the entrance and

develonment of the pathogen.

Table 1. Chart showin; continous treatment and
treatment nlots with ViTlC ies and con

.(D '_Jo

f1

Number of nlants and concentrations of E. a. 1192 F used in
Continous treatment groun.

 

 

Cons. ll92 F Varietv——— Juniter ”ides Nentune Vulcan

1—16,875 25 2h 26 23

1-33,?50 23 2E 25 23
—67,FOO 21 26 25 21

 

Number of plants and concentration of E. C. 1182 F used in
interrunted treatment group. ’

 

 

Cone. 1192 F Variety Juniter ”ides Nentune Vulcan
1-16,€7, 25 2h 25 23
1— —33,750 ' 23 26 25 23
1-67,?00 22 27 25 21

 

ests of Growth hormone Action

It had been rencrted in the literature by Stoddard and
Dimond (19) that plants treated with E. C. 1152 F nroduced
noticeably denser growth than rlants in untreated nlots when
annlied to the carnation vari ety, Miller's Yellow. As a measure
of this hormone-like effect, the aoove autho ors used the nurter
'of " reeks" or terminal growins ooirts ner stem as their criteria. .
Treatments of four carnation varieties revealed no hormone or

’5

Formative effect under the conditions or this exrrs - e.+ The

following chart lnllCu+€C the averace nu ber of "breaks" ner nlant

in each of the treated and check plots. There is no significant
difference bot:eon the nunber of "breaks" or terminal growing
points in the treated plants as compared to the number in the
check plants. There was consistently greater number of breaks

in the check plants in the variety; Jupiter than there was in the

treated plants of the sane variety.

Table 2. Average number 0? breaks per plant from a single low pinch.

 

 

Treatment Vidas Neptune Vulcan Jupiter

A+ a» A B A e A a
1—16,875 3.b5 3.0 3.85 h.3 3.8 5.16 3.3 h.1
1-33.750 3.u2 3.63 ‘ 3.66 h.3 ' h.2 3.8 3.5 h.7
1—67,soo 3.81 3.b . ' 3.52 3.6 n.03 L.1 3.7 b.§
Check 3.18 3.bh 3.56 3.9 3.5 3.7 3.6 b.2b

 

A* --Treated and inoculated plants

2* —— Untreated and inoculated plants

Check - Not treated er inoculated

Fertilization of the plants, which were kept in 3—inch for 125 days,
consisted of periodic annlications of Plant Marvel so as to maintain

soil analysis by the Spurway method as follows: hOpnm. nitrogen;

Spnm. phosphorus, and he pom. potassium. The pH was kept at 6.5.
Method of Recording Disease

The "culturing" method as carried out by several
commercial carnation pronagators is as follows: cuttings or shoots

are removed from mother block stock plants grown especially for

t4
‘ ."l

propagating purposes. The propagator does not use a knife in
taking cuttings, but places a piece of tissue paper between his

thumb and index finger which prevents his hands coming in contact

U)

with the cutting, this lessening the pos iiilty of sp

‘1

sadine any
disease org nisnt which might he present on the plants. The cuttina
is snapped off with the thumb and index finger and a new piece of
tissue paper is used for the next cutti 3. After the cuttings have

3

seen taken, they are rerovei to a sterile room where trained
women workers remove approximately one inch of stem from the base
of the cutting.

The sten section is then cut into four equal parts, and these
are surface sterilized by imrersion in mercuric chloride and then
sodium hypochloride. Scalpels and forceps used in the cutting of
the section into pieces are dipped into alcohol and flamed after
each Operation to prevent contamination or transfer of disease
from one cutting to another.

After the four stem sections are surface sterilized, they are
plated on Petri dishes containing notato dextrose agar. Usually
two separate plant stem sections are placed in one dish. The Petri
dishes are incubated at about 300 C., for a period of seven days.

h
1*

At the end of this time, if there is no evidence 0. Fusarium myselium
or spores or of bacterial wilt colonies, the correspondingly laheled
cutting from which the sections were taken is rooted in an isolated
area in the range, and later grown into a disease-free stock plant.

It should he mentioned that while the stem sections are being

screened for disease, the remainder of the cutting is separately

16.

heat—sealed in cellOphane and placed in a refrigerator at h or 50 C.,
during this time. The materials and tools used in culturing are
shown in Figure 2.

From his method of culturing, mother blocks of thousands of
diseaseufree stock plants are ev<lved from which come the finished
commercial cuttings. These mother blocks of stocs plants are
carefully maintained by workers who do not come into contact with
other carnaticns in the remainder of the greenhouse. Cultural
practices are extremely clean so as to prevent entrance of any
pathogens to the blocks, and steam sterilization of all tools and
equipment is standard procedure.

The author adopted much the same techniques as described above
for positive counts of diseased plants in the inoculated plants, with
the added precaution of dry sterilizing the issue in rolls for 2h
hours at 1800 C. Visual observation of disease symptoms also were
used together with the "culturing" method to obtain the percent
total disease. Fusarium was Considered to be present in the plants
if one section out of the four plated showed mycelial growth in
culture. We sugar was used in the potato dextrose agar to allow
for fast macrOconidial formation which would facilitate the
identification of the pathogen.

Stem sections were take- January 1?, and 13, 1952 from all
inoculated plants which had not shown symptoms by this time. In
the varieties, Jupiter and Vulcan, symptoms were readily evident
beginning the last week in December, 19Fl, and much slower to show

in the varieties,‘Vidas and Neptune. Periodic platings on potato

k

 

in
used
materials

U u t
S L
°ectio

Stein ‘

«j c\;:\qq_ ." r98

- " 0.0“ L

- 1’le

W'quasuo

Ola..-

«on t.
y Titn
Rh to tne C,

basal

“+9“ Sectlpn

 

' 313.
isolatio,

17.

dextrose agar were made on plants sho owing symptoms to check

'Jo

visual recordinq of d seased plants.

Tn general, symptoms of Vusarium wilt cor formed to those
described fully by ‘ickertcn (2), that is, there was u mjl teral
develonment in the tops, with a yellowing of the stem internoies
on the infected s: de, a yellOWinc followed by witherinx of the
leaf bases and a yellowine of the midveins 01 these leaves to the
tin. radually the infected leaves became yellow, no.
side at f‘rct, and finally withered. These symptoms slowly progress
up one side of the rlant and into the lateral branches and shoots.

One of the first si me of the disease, which may not as
accompanied by yellowilg, is a definite outward curling of the
terminal shoot which is infected. Chlorosis usually follows and
the curling becomes very pronor.ced. In the vascular system there
is a distinct discoloration which gradually gives way to a dry,
shreddy reddi W5 rown rot. This progresses up the nfec ted shoot.
Pcots usually are in tact thrc ghcut the life of the plant, hut

‘!

some rotting does take place, especially in the roots where the

(L ..
kl)
93
m
(D
53?"
(D
r-v
(N
8.42...
o
C'"'
(L
L3)

ve rotting usually takes place in the

roots after the disease has almost completely killed the tons of

Flower Production from Diseased and Use lthy Dlants

To obtain an indication as to possiole effects of Fusarium
upon the production of cut flowers, it was decided to bench the
plants out of the 3—inch pots, and keep out flower records

according to a standard grading method. The Cffi'weight ET3diU§

system nromsed by Post (18) was used.

Tefore the soil in the bench was steam sterilized, five
pounds of superohosnhate, five pounds of calcium sulfate, and one—
half pound of muriate o? rotash was added to the soil.

Pecause of losses from Fusarium, an irregular numher of plants
was benched, thus making the results 0? doubtful siyniiicance.
The CS? method 0? grading cut flowers is based on stem length and
flower weight which includes the stem. The followinf table after

Post (18), shows the nethod of grading employed with carnations:

an '51! r-n nv no» V:
u". . 12A 1. IL. ' 31-25311")

 

 

Weight per flower l"Tinirrmrn stem
grade in ounces lensth— ins.
Special 1 and over 2h"

Fancy 3/h to 1 an"
Extra 1/2 to B/h 13"
re. 1 1/h to 1/2 12"

No. 2 ' Less than l/L l2"

 

Table hree shows a record of average cut flower nrcduction of all
four varieties. From a total of 239 slants benched, Fl were
untreated and uninoculated choc? nlants, 109 were treated and
inoculated, but were healthy, 35 were diseased checks, and 15 were
diseased nlants taben from the treated and inoculated nlots. The

average nucber of cut flowers rer nlant is accieved hy dividing the

nuaber of slants into the numher of flowers cut in each groun.

1°.

Table 3. Average number of fTowers cut regardWOSC of yrade .
from various treatments. All varietles are *noluded.

 

Check 2.72 Tlowers per olant
Check diseased 1.22 " " "
1-16,875 2.8L n n n
1-33,7F0 2.28 n n n
1-67,;00 2.17 v n "
U‘seased

1—16,87§ 2.0 n n "
1-33.750 h.o " n «
1-6?,SOO 2.0 n n n

 

There was no relationship between grade of the flower and
diseased or healthy rlants, since reor<eentative graies were ’ound
almost to the same extent in the diseased nlants as they were on

the healthy cheek plants.

20.

~ 1-) ‘UT‘ r4-‘T’T‘h'f RESUTTS

Counts of diseased plants in the various treatments were a
combination of observations of visual symptoms of Tusarium wilt.
substantiated by periodic isolations, plus stem section isolaticns
from all symptomless ino culatei slants, as oreviously described under
Taterials and Vethods. Since no chemical test on olant tissue that

-stantiates the actual oresence of a chenotherareutant is known
at t1e present time, the criteri_um for a551ming that the material
is a sorbed oy the plant 5 must necess ari y be its activity in the
host or cathogen.

Table b shows the results of the inoculations in percent of
disea Me olants in the interrunted treatments.

Tasle h. Percent of disease in 288 plants in which treatments were

stomped at tin of inoculation.

 

Gene. 1192? Juniter Nidas Vulcan Neptune

 

1—16,87§ 32.0 33.3 21.7 t0.0
1—33.750 A3.b 3R.h 21.7 36.0
1-67,;00 10.9 33.3 1L.2 2h.0

Check 71.05 77.h 26.6 .8.08

 

The average nercent of disease for the groun of 288 plants shown
in Table h was 31.9.

Table 5 illustrates the average percent 07 iisse P in that

r“

n n . . . 3 .
it no sulatei lan s in which the treatments 1v HP contin—

h)

"I
:rcun o-
s.) -

ued for eighty days past the date of the first inoculation.

21.

Table F. Percent 0’ disease in 298 nlants in which treatments were
continued after inoculation.

 

 

1.1
(L
C.)
U)

Cone. 1182? Juniter V' Vulcan Nentune

 

1-16,875 20.0 16.6 0.0 0.0

1-33,?50 17.3 16.0 0.0 0.0
1—67,§oo 1h.2 16.3 0.0 0.0
Check 71.05 77.h 26.6 68.08

 

The average percent of dis ass in the above plot was 8.3.

L2.

The significance of this is doubtful since no infection was obtains
in the two varieties, Vulcan and Neptune. Possible causes of this
are noor techniques, incubation time was too short, symptoms were
masked by some internal factor, or the chemotheraneutant nrovided
ahsolute rrotection against the disease's entrance.

The average percent of disease in the check inoculated nlants
was 6b.].1,‘ 5-. .E",.‘I.r;.

The check plants which were dinned in distilled water and

were not treated or inoculated did not become diseased.
Symptonless Diseased Plants

A record was kent of the varieties plated for attempted
isolation of Fusarium from svmntomless nlants which had heen
inoculated. All stem sections were taken from annarently healthy
plants which showed no symptoms of Tusarium.

Table 6 shows that the varieties, Vulcan and Neptune might
possihly as syn tonless carriers of :usarium, since a large percentaes

o: the stem sections taken from apparently healthy nlants were shown

infected with the disease when nlaced on notato dextrose agar.

iable 6. Percent svrntomless nlants shown by stem sections
isolations from annarantly healthy slants.

 

. . ‘ .
VarIEty Percent Diseased

 

Jupiter 22.2
Midas 33.3
Vulcan .2.5

Neptune 73.03

 

.'0

DTSCUSSIOH

E)
3

es its the failure of two varieties in t‘e continuous
treatnent lot of 29s nlants to hecwe in_ected, there is an

indication here that the chemotherareutant in ouestion is exerting

two effects —— one noon the host and another on the pathogen.

f?i_nce the int Hr un tei treatment showei a reduction of disease from
6h.1 neroent in the check-inoculat A nlants to 31.9n nt in the
interrunted groun; and since the ch nWtier aneutant was not annlied

after the time of inoculation, the reduction in disease must be a
result of the action of T. 3. 1182 F unon the host and not unon the
nathoeen. Tn the continuous trea,ment plot, if we disresard the
two varieties which failed to hecone infected, it could he noted
that the total percent infection in Juriter and V'idas is only 16.-,
a still greater reduction fron.the nercent disease in the interrupted
treatment. This couli indicate the t l) T. C. 1182 F exerts some
direct fungotoxic1ty, 2 the material is keinin" t4e nlant in a more
or less constant state of resistance to the entrance of the nathogen

by its constant n esence in the nlant ,3) or a carbination of tee

.4.
O
’3
D
H
.1)
C I“
PM]
:3
70
T‘
H
O
4
C T‘
L 3'
.7)
,4.
J
(J)
'"J
1‘.)
V)
(D
'j
T)
’3
CU
. J
c f-
5.13
’2
\D
O
'J

tvo pee siiilities
the continuously treaeei ~lants below that of the interr runted treat—
ment.

These experiments are ed on a ternerature of 60° F. Since

a hieher nercent of infection by Fusarium occurs at temneraturec

2L.

of 75 to 900 T., when inoculation is through the roots, it might
be conjecturei that in the treated clotc, the decree of control
might not have been so great at hirhcr tennerntures.

Cut flower reccris iniicate that Fusnriun—infectfd nlants
produce a much lower nurbcr CT Flowers than do disease—Tree
slants, thus nointins to the desirability of "culturei" cuttings
in the carnation indvstry. This reduction in cut flower production
is probably a result of a decreased oroduction of vegetative and

flowering shoots.

M
U1
0

." .‘TfiT T‘C‘ 'T‘r‘WUF‘
UC. also”. LL15.)

Tron the nrecedinq exneriment, suggested recomncndations for
the control of Tusar um wilt of carnation can be evolved.
Primarily, the procurement of disease-free cuttings fro”

renntable carnation nronagators is of great innortance. If

r 5

—nrona2ated, cuttings are free from.?usariun
wilt, the other tro crecautions exert much more influence unon
the maintainance of the disease—free state of these nlants
ttroughout their life in the greenhouse.

Assuming that the cuttings are disease-free, tnen strict
sanitary measures must (3 ennloyed in the rooting, potti.g,
benching ani cutting of the nlants and Flower For their entire
life in the greenhouse. These clean habit? of growing will not
only hinder the entrance of Fusarium into the nlants, but will
also prevent its snrcai should it enter in snite of these
precautions.

Lastly, in combination with, and sunnlementing these two

practices, a chemotheraneutant, such as E. 3. 1182 F, couli be

-4

annlied bi-‘eekly throxhhout the life of the plants un to and

2

including the flowerinw st«.e, so that the nlants are constantly

(J

L)
C:

in a state 09 re

U)

istanoe to the entrance of the aisease organism.

While this material is not the final answer to control of "usarium

-‘

wilt of carnation aha will undoubtedly be surerseied by newer, more

powerful chemotheraneutants in the future, it shows sufficientlv

" u

SZ'OOj. 1985111t5 t0 ‘4‘731413'37‘1t its US?! in 0("1WWF‘Y‘r‘W'Q-l r-n‘r'nntinn nvonnhmwcz‘c

26.

C? "I “
\JUT‘ ; nifty

A chemotherapeutant, h—chloro~3, S—dimethyl phenoxy ethanol,
was annlied in soil solution to tour varieties of carnations in
an effort to control Fusarium wilt.

Visual symptoms of the disease caused by Tusarium dianthi,

 

plus an adaptation of the commercial method of "culturing"
cuttings were used to arrive at the following total nercents
disease: check, 6h.1; treatment stonned at the time o? inoculation,
31.9; and continuous treatment, 8.3. Two varieties failed to
become infected after inoculation in the latter groun.
Cut flower records of flowers cut ner plant, based on the
CSfl'weight graiing system as annlied to carnations indicates that
healthy slants cut-nroduce diseased olants by a noticeable margin.
The data shows that 2.72 flowers oer nlant were cut from healthy
check plants, while only 1.22 were cut from infected check plants.
The disease anoarantly lessened the number 0? flowerine shoots
produced.
as use 0? disease—free uttings and strict sanitary measures
in the growing of carnations is strongly recommended. ii—weekly
aoplications of one part 3. C. 1182 F in 67,500 parts of water

have been shown under the conditions of this experiment to nrovide

a sizeahle reiuction in Fusarium wilt.

l.

9.

"'7 I “I 'Y'" '7' C 57.7"
LLJJ‘LLCLLZLL LLL

Ark, P. A. Chemical eradication of crown gall on
almond trees. thtopath. 51:956-957.
lyhl.

Eickertcn, J. M. Fusarium wilt of carnations
caused by Eusarium dianthi Prill. at Del.
N. Y. (Cornelll Agr. Exp. Sta. Eul. 788.
pp» 1-51. 1942.

 

Bolley, H. L. Potato scab and possibilities of
control. N. Dakota Agr. Exp. Sta. Eul.
A. pp. l-la. 1891.

Bouchereau, P. and Atkins, J. C., Jr. Studies on
the action of copper seed treatments for
water-planted rice. Phytopath. u0:3.
19500 (AbStraCt).

Brown, J. C. Cytological effects of penicillin
and streptomycin on crown gall. Phytopath.
58:5. 19h8. (Abstract).

Brown, W. Stem-rot and wilt of the perpetual
flowering carnation. Sci. Hort. 6:95-96.

1958.

Chapman, R. A., A. E. Dimond and E. M. Stoddard.
Assaying chemotherapeutants in the
greenhouse. Phytopath. h0:h. 1950.
(Abstract).

Chapman, R. A. Relation of specific chemothera-
peutants to the infection court.
Phytopath. 41:6. 1951. (Abstract).

Dimond, A. E., Z. M. Stoddard and R. A. Chapman.
Chemotherapeutic investigations on the
common bacterial blight of beans.
Phytopath. h2:72-76. 1952.

On the stem rot or wilt disease

10. Dowson, W. J.
of carnatiors. Ann. Appl. Biol. 16:

261-280. 1929.

Cure of crown ga
Phytopath. 38

11. Hampton, J. E. 11 with anti-
biotics. :11. 19u8.
12. Eorsfall, J. G. Combating dampingeoff of
tomatoes by seed treatment. K. Y.
State Aer. Exp. Sta. Bul. 586. pp. 1-22.

1950.
15. , and A. E. Dimond. Plant Chemotherapy.
Ann. Rev. Microbiol. 5:209-222. 1951.

a remedy for or

V ' Bordeaux mixture,
Rhode

1h. Kinney, L. ..
preventive of the potato scab.
Island Agr. EXp. Sta. Bul. 1h. pp. 131-
186. 1891.
15. McNew, G. L. and N. K. Sundholm. The fungicidal
action of substituted pyrazoles and
related compounds. Phytopath. 59:

721-751. 1919.

16. Newton, R. J., V. Lehmann and A. E. Clarke.
Studies on the nature of rust resistance
in wheat. Can. Jour. Res. 1:5-35. 1929.

17. Nugent, T. J. and H. T. Cook.
seed disinfectant for control of black
1939-

rot of kale. Phytopath. 29:19.
(Abstract).

Weight grading of cut flowers.
1918.

Chloropicrin as a

18. Post, Kenneth.
Flor. Exchange 110 (2):23-25.
The species

Y. Hansen.
Amer. Jour. Bot.

19. Snyder, W. C. and H. .
concept in Pusarium.
27:6h—67. l9h0.

The chemothera—

20. Stoddard, E. M. and A. E. Dimond.
peutic control of Eusarium wilt of carna-
1951.

tions. Phytopath. hl:557—5b0.

21. Strong, F. C. and D. Cation. Control of cedar

22.

25.

rust with sodium dinitrocresylate.
Phytooath. 50: 983. 19h0.

Sturgis, W. C. preliminary irvestigations on a
disease of carnations. Conn. Agr. Exp.
Sta. Ann. Rept. (Part 5) 21:175~181.

White, B. L. lhe wilt disease of the carnation.
Jour. Pomol. and Hort. Sci. 7:302-325.

1929 o

Wickens, C. M. Wilt, stem rot, and dieback of
the perpetual flowering carnation.
Ann. Appl. 5101. 22:650-683. 1955.

Yarwood, C. E. Therapeutic treatments for been
rust. Phytopath. 57:2h. 19h7.
(Abstract).

 

_

 

 

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