X My: .: «a QC mil. Hm . rm. .7! Quo’r.‘ pr U a: I In“) a»? ‘0 t ‘ "EEEVE ~§ney FT 7.4 .0 | ‘ game ~ .qu F‘t mum...» . I .3. o t . o Wm H. mm. .. .6 flaw coo mks” a. .nd mu. m .o 2». .«mrtt K 22. y“: “.3. l P‘ r 3“: I!!! 3. nu,“ ‘n' InWI é‘u v. ‘0. .' y f F. ' «war... 1.... fl (a M!” u.“ .m u... x» 3.. w... mm ... a C " t k: m: m- ‘2 Hg2:__:3::3:::33;3:; mmm h I, . THESIS i I LIBRARY Michlgan State University 3y Arthur J. Cine 3 An investigation was coniuctei to establish if there was a difference in the rootability of Taxus clones ani if this difference could be relatei to anatomical or sexual differences of the clones Seven clones of Levu), plus male and female 2. cus- piiata expanse Eort. were propagatel by tip cut tings :iurin3 the winter of 1963-193#. The cuttings were placei in a sand . 4.. U media and kept under intermi ent mist du ring t‘r e ex oer 1men Anat mical studies were maie of the stens of the rootei ,J cuttings during the winter of 1963. The transverse sections of stea were examined for the number of fiber bands, and the 3 1' 1... .0 v 4— - '. 1 ' tnicsness Oi oarl b0 @001 in eacn section. H) Results 0 this studr show a definite difference in the rootabilitv of Taxus clones. T. seiia 'Jetfielo' showed the hi3hest root ability and E. cusoiiata 'Hana' exhibiteu the lowest. Cuttings take n from male 2. cusgiiata exgansa rootei more reaiily than cuttings taken from female plants. The anatonical stuiies shorei no relationshio between structures studied ani rootaLilitY. ,. I O 0 VJ t J (.J L J p v f L3 1") r }_ 3 I ‘-a 31% ‘_ cf K H [JJ U] H *3 :1) m (—c U) U ”x 0] Arthur J. Clney A T€SSIS Submitted to Iichigan State University in partial fulfillment of the requirements for the degree of '."" 1 “‘1 :41 D erartment of horticulture H \C) O\ \J‘x J .‘Wffi‘fi ‘31 a. J—J.'./’ V-- >13 .3 TO hi I. Harriet C. and Walter F. Olney Ho Ho ACKNOWLEDGEEEN;S The author wishes to express his gratitude to Dr. Harold Davidson for his guidance and assistance in the conduction of this study. he is also grateful for the help and advice given to him by Dr. Edwin D. Carpenter, Jr. and Dr. Arthur E. Mitchell in the anatomical phase of the study. Thanks are expressed to Dr. Roy A. Hecklenburg and Er. Clarence E. Lewis for advice and suggestions. The author is grateful to Hidden Lake Gardens for the financial support of this study. 111 A ~11 .q_r~‘-~.-11-~~vr-v-1 (-1 m3'fi‘r'n ----._—_._,.. \u- \o \-'.. an“. .~ J TR‘TI‘F‘ ' n 1\T",~flr‘fij- Afi' .hm‘ d. -- I " ' v--vv....Lv.-oooooooooooooooooooooooo ‘L)~"'-,rT1-v-‘nv--:1~f‘n T!“ 3.? ‘mTT')'_‘ -_4----.1._._-.- '-'_...-J---¢_v_~—Jo000000000000000 5...-J— ...-,. -. ,..+.- V. ._L. ..,, 4.0- -. ‘n‘h;:—Vb -‘U 3.;. re;“ La'lOLA L’O LOOH’l;-:‘/. 3" . -4. .L. . .. ‘qu “51 ., __ I} 1 .11. ‘rfi ‘ 4L4; I _x_ -'._1~.g L/O p... Jr}-.. - ' 1051...... m,fiVP‘1!--‘ ._~.—----a.000000.000000.000.000.000 *— ‘fi ‘ r"'\-'\ out-.- .— '"i'1'"""~”fi ' T' "~""\f\l‘|"7"“\’7“"—‘ " ' .l " I . .o.‘.04-I--.b-h.&_ ‘5 .~'\rv.—_o\/.-~—J‘J.O........... .mnpwawme 'T' ‘ _._---v.-~/ *OOOOOOOOOOOOOOOOOOO v—1 _ o T 4"Q«'\‘lfil“‘¥(\1’7+— T: 4.4-‘u- «‘v~—-~/ _‘.oooooooooooooooooo yr. 9 g m V o .. .~ ~ r .,-—.,-._.\-'-,l ,~-u_r.¢ histologicil low--- \2 -o........ vs-.. 4... . .D .1 ° V" 4 . n ma n <3 [3-73 BJM_..‘LLV«.L/l '11 O; .-_l_.-'v.;ooooooooooo -oa—wv—.-‘~JOOOOOOOOOOOOOOOOOOOOOOOOOOOOO «~pi‘.":‘~qi-,AA.A‘q L . ‘ " I HA-IJVJ. ---~_,-../ 10000000000000.0000. ”7""3p-a ‘7‘. r531 4" T .4“ vi,..~\..-..LU -oooooooooooooooooo ."tont'quiNH "O'M‘ ‘r‘r‘ ' -LsApb'J\.-A._L'-. -.b~2~x-_v00000000000000 “Trflvdv'rhv ~‘.~~."~ls.‘.JJ.L.K.w--OOOOOOOOOOOOOOOOOOOOOOOOOO fiTT"“' ‘ .-‘_- r .\ »| J~-«--.~‘-ococoaooooooooooooooooooooooo :I“LTo~nzuwv ~‘A—Ad‘n-é—O...0.000.000.0000..0.0. i V \ _-) \J.) ”J H “Q (‘0 C) I‘) O P) \D \t) ( LIS r‘ OF T'A‘73I33‘S .A..3‘ FIJUTES SELL"; OF TA‘CUS CL LESS TAKUS CLONES USE) IN ELI-{£3 S‘II'LELT'I‘ II THE EOOIIICG Il‘T‘ICES OF LL” VS. F aJU‘I'Ti‘TGS OF I'd-{1:3 cusaidata expan 3001 WI: 1N33r3s OF r33 ”i’“‘ CLoxss 3.1331315? BELLE TITO 3400) OF :Li: 73 cus_‘._.')idata xpansa m airIo OF 3Ai£ ro woou IN FACES cLOflES STEM 371033 SLUTIOI OF TAT-{US media 'Ia.tfield' L31 20 21 22 23 21+ INTiOUUCTIOH Plant propagation has been defined as "the controlled reproduction of plants by man to perpetuate select indiv- iduals or groups of individual plants which have specific value to him." (14) The propagation of plants is the starting point for all crop production, and as such is of fundamental importance to the field of horticulture. Plant propagation may be easily subdivided into two major divisions: sexual reproduction and asexual or vegetative reproduction. Due to the fact that many ornamental species have seeds with complex dormancy mechanisms which hinder germination, asexual reproduction is employed with these plants. Asexual prop- agation of ornamental plants on a commercial scale is done cheifly by means of cuttings. Since a greater uniformity of plants can be obtained, and a larger number of new plants can be produced from a relatively small number of stock plants, cuttings fits well into the scheme of mass pro- duction. Many factors affect propagation of ornamental plants by cuttings. One of the most important of these factors is the inherent rootability of the cuttings. Two of the inherent factors which affect rootability1 are sexual and anatomical differences. Beakbane (4) has reported that the anatomy of a cutting may affect its rootability; Snow (26) observed differences in rooting associated with the sexual character of red maple cuttings. Taxus, a dieocious plant, (1) occurs commercially in 1 Rootability - the tendency of a cutting to form roots. 2 varieties of mixed sexes, and in clones which are either male or female. It has been observed that cuttings from different clones of Taxus vary in their degrees of root- ability (30). It has been the scope of this study to attempt to establish if there was a difference in the rootability of Taxus clones and if this difference could be related to anatomical or sexual differences of the clones. A REVIEW OF PEdTINENT LITEingEE AFATONY IN HELATIOH TO ROOTING AND VIGOE Work which started in 1912, at the East Kalling Research Station, England, led to the development of a series of clonal apple rootstocks (14). The different rootstocks were observed to influence scions grafted upon them by producing trees which ranged from large, vigorous trees to plants which were definitely dwarfed. Seakbane, et. al. (2) studied the anatomical structure of the East Halling rootstocks in an attempt to correlate anatomical features of the plants with the rootstock effect. She found that in dwarfing rootstocks that the proportion of bark2 to wood was very high, while in the vigorous rootstocks the proportion of bark to wood of the roots was quite small. On the dwarfing rootstocks the roots were observed to con- tain a large percentage of living cells, while the roots of the invigorating rootstocks were found to contain consider- able lignified tissue, chiefly without living cells. Beakbane (3) also found a relationship between the proportion of fibers in the bark of apple roots and the vigor of the plant. The amount of fibers in the dwarfing rootstocks was found to be low, while in the more vigorous rootstocks the amounts of fiber generally were quite high. In plum rootstocks there was a relationship between the anatomy of one year old stems and the ease with which cuttings could be propagated. Root- stocks which had groups of primary phloem fibers separated 2 Bark - In this study the term bark will mean all structures outside of the cambium. ’2 J h by large areas of living cells had the highest rootability; other rootstocks which had stone cells, or sclereids separ- ating the primary phloem from the secondary phloem produced cuttings which were very difficult to root. Beakbane (3) studied the anatomy of rootstock stems to which Lane's Prince Albert had been grafted, in an attempt to correlate the rootstock effect. The stems of the dwarfing rootstocks showed a higher bark to wood ratio than the more vigorous rootstocks. Nosse (20) in a study of unworked plants also found that the percent of bark was greater in the stems of the dwarfing rootstocks. Santamour (24) studying anatomical characteristics of poplars reported that the percent of root bark appeared to be a definite clonal characteristic. He examined roots and stems of one year old cuttings taken from twenty-five clones in two, nine year old plantations. He found that the root bark percent of the one year old cuttings was significantly correlated with diameter, height, and volume of growth of the clone. It was found that clones with a low percent of root bark were vigorous, while the clones with thicker root bark tended to be less vigorous. Santamour also found a positive correlation between the amount of fibers in the root bark, and five growth variables; he found that the clones with the greatest area of fibers in the root bark were most vig- orous. Other anatomical characteristics examined in the root, but which were found to be nonsignificant between clones were: percent of vessels in the wood, the size of the vessels, and the percent ray tissue of the wood. 5 Beakbane (4) reported that the sclerification of the primary phloem of one year old stems was closely related to rootability. Poor rooting varieties exhibited a high degree of sclerification in the phloem, while in the more easily rooting varieties the primary phloem parenchyma was living, even in the transitional stage to scleroids. The phloem rays in this type of plant appeared to terminate against living tissue, while in the plants which were difficult to root the phloem rays usually terminated against heavily lignified fibers and sclerids. These differences in the inherent sclerification of the primary phloem has been found to be quite stable for similar material. Beakbane also suggested that the major cause of poor rooting was not merely a case of lignified tissue mechanically blocking the devel- opment of roots, because in many plants root initials do not even form and try to force a passage through a lign- ified layer. She suggested that it should be entirely poss- bile to predict the rootability of cuttings taken from a plant chiefly by observing the apparent structure of its young stems. Killer (15) studied the effect of growth regulators on the origin and development of root primordia and callus tissue in Taxus cuspidata. She describes the anatomy of the basal region of a cutting taken from one year old growth. The secondary xylem was observed to consist of uniseriate rays and tracheids having bordered pits and being two to five cell layers thick. A primary xylem which contained pitted, retic— ulate, spiral, and annular elements. Since the cuttings had 6 been taken during the winter months the cambium was dormant. The author noticed a zone of immature elements of from three to four cells wide separating the xylem and phloem. "The phloem consists of alternating tangential bands of sieve cells and parenchyma and of phloem rays. The phloem cells at the periphery of the stele are crushed against the tang- ential band of parenchymatous cells approximately three cell layers wide, which surr unds the stele. These cells are considered to be the inner cells of the cortex." A starch test was conducted by the author to determine if an endo- dermis was present in the cortex. The test proved incon- clusive. Hiller found the cortex to consist of from one to three outer layers which contained starch and chlorophyll, and the remaining portion to be composed of thin walled parenchyma cells. The epidermis was covered entirely by a thick cuticle. Taxus cuspidata does not have preformed root initials, so that in a cutting the root primordia arise in the secondary phloem in connection with a vascular ray, or in the wound tissue at the base of the cutting where the primordia arise within the undifferentiated phloem which has no vascular rays present. Chang (7) studied the bark structure of Taxus brevifolia, and found that fibers were present in the outer part of the inner bark. The fibers usually occured as tangential uniser- iate bands WhiCh alternated with bands of sieve cells and parenchyma. Esau (12) states that fibers are present regularly in Taxaceae phloem. 7 SEX RELATED TO PROPAGATION The sex of individual plants or clones and the presence of flowering buds upon cutting wood have been found to affect the rootability of cuttings. Snow (26) found that male and female red maple trees from various selected greenwood cut- tings differed in their ease of propagation. In a comparison study it was found that cuttings which had been taken from female trees did not root as well as cuttings which had been taken from male trees; he also noticed that some of the female clones exhibited a high frequency of root formation, while others rooted comparatively poorly. Egerton (11) followed up this study, and reported that cuttings from female trees which had borne a light crop of fruit rooted equally as well as cuttings taken from male trees. He suggested that the trees which bear a heavy crop of fruit tend to be low in carbohydrates, and thus root poorly. A study con- ducted by O'Rourke (22) on Blueberry, showed that cuttings which bore blossom buds rooted poorly in comparison to cuttings taken from vegetative wood. Chadwick (6), in a discussion on the use of flowering wood versus vegetative, states that the apparent effect of flower buds on cuttings is to increase the rate of respiration markedly over that of the vegetative buds, such that considerably more stored food is converted in the rooting process. This decrease in the stored food supply of the cutting appears to be a contributing cause of the poor rooting of cuttings bearing flower buds. Neal, et. al. (21) in a study of Ilex verticillata took 8 cuttings in June; nearly all of the male cuttings rooted, compared to approximately fifty percent of the female cuttings. Cuttings which were taken in the middle of July showed only minor differences in rootability between the sexes. Neal suggested that the apparent difference between the male and female rootability of cuttings taken in June was due to low levels of carbohydrates in the female plants, a result of heavy cropping the preceeding year. By July the carbohydrate deficit had been returned to more normal levels, and no apparent differences in rootability were observed between male and female cuttings. DeBoer (10) found that ghododendron cuttings rooted better when the existing flower buds were removed from the cutting, than when they were allowed to remain; she suggested that this result may have been connected with the plant's natural hormones and inhibitors. According to Bailey (1 Tarus sap. are evergreen trees 9 or shrubs which have linear two ranked leaves. Flowers are dioecious, and occur axillary, when appearing in early spring. The seed, a hard nut, is enclosed on three sides by fleshy red fruit, and requires two years to germinate. Presently Taxus is considered a valuable plant in the nursery trade. Keen and Chadwick (13), Davidson (9) and Hyman (23) have reported on the sex of the different clones of Taxus as they occur in the nursery trade, at the Seacrest Arbor- etum and the Arnold Arboretum. Table 1 lists some of the etter known clones by their sex. Keen and Chadwick (19) 9 also have reported that they have observed several sex rever- sals in T. cuspidata where several male plants have been observed to possess one or two female branches. Except for special instances, most of the clones of C3 Tarus are orooa3ated oy means of cuttings. Keen (17) pre g '\ ‘... r-u ented a good review on the propagation of warns which covers some of the meth ols used for propagation other than by cuttings. Chadwick (5) found that the best medium in whicL to root %.:us cuttin s va ed during the year. feat moss produced good results from July to November, but sand was proven a more satisfactory media after: ovember. Vermullen (23) and Wells (29) also reported sand to be the best media for Tarus propagation. Commercially mavus cuttin.3s are taken from ‘ovevocr to January, and propaga ed in the greenhouse. Wells (30) reports that the larger the outtin3s taken, the easier they will root; thus economics determines the size of the Tarus cutting. Teen (1?) reaorts that cuttings taken from Tatus continue to produce the type of growth characteristic of the position of the parent pla it from which the cutting was taken. Cuttings taken frozn the side of the parent plant produce one sided plants, while cuttings which were taken from the too portions of the plant produce uiri.Q ht plants Esper (13) found that a bottom heat of 65-700 F, and an air temperature of 50—550 F produced he best rootability. Several authors (30, 2 ) have reldorted an increase in rooting per- centage using growth suoste .nces on cuttings. Wells (29) reports that the use of growth substances 1L the form of commercial talc dusts to be a common practice among nursery- 10 men. Photoperioiic effect on the rootability of Tarus cus piL ta has been studied by Slyier (2 7); he found that the lateral bud growth on cuttin.3 s Iias in ibited by a short day of eight hours, and was stimulated by long da Tys of ei3hteen houis of li3ht. Althou3h lateral bud growth was affected by the photoperioi, no sijn -ifi cant effect was found on the root- ability of file cuttin3s. Clonal differences in tne rootability of EEEEE have been noted by several authors (23, 30). Host of the data are given in percent of the cuttin3s rooted; this does not indicate the true rootability of the cuttin3. O'Rourke (23) recommended the method of using ranks to score rootability of cuttings. Wells, (30) usin3 this rootin3 index, gives a more accurate measure of the rootaoilitv differences between clones of TaXus. From his data it can oe seen that T. media '?ichs' and 2. media 'Srown' are poor rooters. Gotta-’5?e Gardens3 has reported that clones of ’1‘. media 'srox-m', T. cu spidata 'Anferson', T. media 'ward', and the variety . cuspidata 'Lana' all have poor rootabili , while such varieties as T. cuspidata and T. media densiformis root easily. Davidson et. al. (9) did a survey of the Hichi3an State University campus and several nurseries in the state and found that male plant of TaXus cuspidata eXpa nsa 3r atly out- numbered the female plants of this variety in all locations. 3 Personal conversation with Kr. Albert Ca rlitZ, of Cottage Gardens, Inc., Lansing, hichig ar - 4-. 11 Table 1: Sex of Taxus Clones Taxus Baccata Sex and Authority ________ 'Adpressa' Female 17, 23 __ _ 'Aurea' Female 17, 28 ________'Columnaris' Fale 23 _______ 'Dovastaniana' Female 23, Hale 17 ________'Elegantissima' Male 28, Female 17 ___-____'Erecta' Kale 17, Hale and Female 23 ___ ___ 'Expansa' Female 17 ___ ___ "Fructu-Luteo' Female 17 ____ ___ 'Glauca' Male 17 _ _ 'Lutea' Female 23 ___-____ 'Hioun' Kale 29 _ __ 'Nisra' Male 17 __ __ 'Pendula' Female 28 ___'___ 'Repandens' Hale and Female 9 Female 28, 17 ____ ___ 'Variegata' Female 28 _______"Washington' Female 17 Taxus cuspidata _______ 'Adams' Male 17, 8 ___ ___ 'Aristocrat' Female 28 ___ ___ 'Aurescens' Male and Female 23 Male 8 __ __ 'EObbink' Female 23 ________'Columnaris' Hale 23 _______"Densa' Female 17, 23 ‘Expansa' fiale and Female 8 Male 28 Table 1: Sex of Taxus Clones (continued) Taxus cuspidata (continued) 'Intermedia' 'Jeffery' ' Elana ' 'Nana Comyaota' ‘- 'Ovata' 'Frostrata' 'Robusta' 'Stoveken' 'Thayerae' 'Thompson' Nedia !-3 51) L r a U) H 9- W 'Adams' 'Andorra' 'Amherst' 'Anderson' 'Berryhill' 'Brevimedia' 'Brown' 'Clifton' 'Cole' 'Compaota' 'Zensiformis' 'Erecta' Sex and Authority Male 28, 8 Female 17, 28 Male and Female 8, 28 Male and Female 28 Female 17 Male 8, 17, 28 Male 28 Male 8, 17 Female 8, 17, 28 Female 28 Kale 28 Female 28 Male 8, 28 Male and Female 28 Female 8, 28 Female 8 Hale and Female Male 17, 28 Kale 28 Female 8, 28 03 Female , 17, 28 Female 28 Female 28 Male 8, 28 Male and Female 28 2able_1: 13 Sex of Tamus Clones_(continued ’_-—'- --.. Taxus Iedia (continued) 'Fastigiata' 'Flushing' 'Halloran' 'Eatfield' 'Fetz' 'sicks yl' 'Hoogendorm' 'Kelsezi' 'Koon' 'Hewport' 'Nigra' 'Ovata' 'Prostrate' 'Stricta Viridis' 'Siebold' 'Taunton' 'Tot-m' O 'Vermeulen' ..—-.o- _- -¢I-‘-. Sex and Authority Male 28 Female 28 Female 28 Male and Female 8 Male 17, 28 Female 28 Female 8, 17 Male 8, 17 Kale 28 Female 28 Female 8, 17, 28 Kale 8, Female 28 Kale 28 Male 8 Female 28 Female 8 Hale 28 Female a d Kale 8 Male 28 UL) I—fi 2 : em.‘94__ e Female 8, 17, 28 Male and Female 8 Female 17, 28 Male 28 EXPE8IHENT I Cutting wood of both male and female plants of Taxus cuspidata expansa was obtained from the horticultural gardens at Michigan State University. The wood was taken on November 16, 1963, from the current seasons growth. This wood was wrapped and stored for two days in moist sphagnum moss at a temperature of 40 degrees Fahrenheit. On the eighteenth of November the 200 male and 200 female cuttings were made and placed in a coarse sand media. The rooting experiment was conducted in a propagation bench of the pit house at the Hichigan State University greenhouse range. A completely randomized experimental design was used with ten replications of each sex, and consisting of twenty cuttings per replication. Bach replication consisted of a row of cuttings which extended half-way across the width of the propagation bench. The cuttings in each replication were placed one inch apart in the row, and approximately two inches between rows. The temperature of the rooting media was maintained between 60 and 70 degrees Fahrenheit for much of the rooting period. In the initial stages of the exper- iment the cuttings were watered as necessary. Due to partial drying out of the media during the weekend of January 19, 1964 it was decided to continue the experiment under an inter- mittent mist. The mist system was activated on January 20, 1964, and remained in effect for the remaining part of the experiment. The mist system was operated on a cycle of six 14 15 seconds on out of every three minutes from 9 a.m. to 3 p.m. ’ daily. On Kay 10, 1964, the cuttings were evaluated on the basis of rootability according to the method of ranks as outlined by O'Rourke and Maxon (23). The results of this evaluation were analyzed statistically by analysis of variance for a completely randomized design. The pur ose of this experiment was to determine if there was any natural difference in the rootability of Taxus clones. Cutting wood from seven selected clones of Taxus (Table 2) was obtained from nurseries in the Lansing area. The wood was taken on November 27, 1963 and stored in moist sphagnum moss at 40 degrees Fahrenheit. The wood was made with slanting basal cuts into uniform seven inch tip cuttings. These tip cuttings were placed in the medium without application of rooting hormones. The experiment was 2 conducted in the pit house of the Michiaan State University C greenhouse range. The cuttings were made and placed in the coarse sand media on Uecember 8, 1963. A completely random- ized design consisting of ten replications per clone with twenty cuttings per replication was used in the placement of the cuttings. The arrang ment of the replications was similar to that of the first experiment, xcept that the replications were approximately one inch apart. The cuttings were maintained under the same watering system, and after January 20, 1964 under the same mist system and cycle as described for the first experiment. 16 T2383 2: Taxus clones used in the Clone 3?; Taxus media 'Hatfield' Rehd. hale Tgxus media 'Nard' Female E?£;s media 'Jensiformis' Male i4xus_:3Lia 'Halloran' Female Taxus media '3rown' fiehd. Kale .gaxgs‘media 'Hicks' Eehd. Female Taxus cuspidata_'33na' dehd. Male Eootability of the clones was evaluated on Kay 10, 1964 using the method of ranks as outlined by O'Rourke and Haxon. The differences between clones were analyzed statistically by analysis of variance for a completely randomized design (16) and differences separated by Dtncan's multiple range test (19). T? :vn. m—rw rTnmv-r-r '\ 711—. RI JLOLOUL VALE-.1 1-3.1-1; I'cJJLf) The purpose of this phase of both experiments was to determine if any anatomical feature of Taxus could be used to account for the variance in rootability which had been observed. After the cuttings from the two experiments had rooted, and were evaluated they were transferred to a cold frame. The media in the cold frame was an equal mixture of sand and peat moss. The plants were maintained under a screen shade and watered every other day. The material remained in the cold frame until larch 19, 1965; at this time plants were removed to the laboratory to be sectioned. Four plants of 1 V each clone and each sex of T. cuspidata expanse were chosen at random from the cold frame to be sectioned. The plants were washed and the distil one half inch of the stem of each plant was placed in the cla ps of an American optical sliding microtome and sectioned to a thickness of 30 microns. The material was sectioned within an hour after being brought into the laboratory, and the sections were started through a safranin and fast green staining procedure as outlined by Sass (25). Stained sections were then mounted in Canadian balsam on standard microscope slides for further examination. EVALUATION or SL 333 On examining the microscope slides, bands of fibers were noted to occur in the secondary phloem. It was decided to count the number of fiber rings appearing in the different clones, and in both sexes of Taxus cuspidata expansa. The purpose of this was to see if there appeared to be a relat- ionship between the number of fiber rings that any one clone contained and the rootability of that clone. In order to facilitate the counting of Ken-a-Vision microscope slide projector was used to project the image of the section, and the count was made from this projectiOI. A 6.5 mm lens and a 10 x magnification was used to project the image. Three counts were made per microscope section and these values were used to find an average number of fiber rings per section. Twenty sections per clone, and twenty of each sex of T. cus- pidata expanse were evaluated in this manner. Analysis of variance was used to test for significant differences in the number of fiber rings. It was decided to determine if the thickness of the region on which the fibers occured varied between T. media 'Hatfield' and T. cuspidata 'Nana'. These clones were chosen on the basis of their rootability, the former being high and the latter low. The microscope slide projector was also used in this determination. The sections were pro- jected as before, and a sketch was made of the image. Nota- tion was made of the ring of xylem and the outer limit of the region of fibers. Due to the irregular shape of the Taxus stem it was impossible to determine an exact center of the section. A method was devised by which a section center was designated. By inspection, a ruler was placed across what appeared to be the diameter of the xylem and a point was placed at the center. This was done three times using what appeared to be different diameters of the xylem. The three points were connected and the center of the triangle was chosen as the center of the section, or if the three points fell in a straight line a point half way along that line was chosen. Three radaii were drawn with a straight edge out through the region of fibers. A measurement was made of the thickness of the region of fibers along the radaii. These measurements were averaged to give an average measurement per section. Twenty sections of each clone were evaluated in the above manner. Significance was tested with analysis of variance. It has been reported by Mosse (20) that the percent of bark to wood could be a clonal characteristic. A study 1'9 was conducted upon the clones of Taxus to determine the percent of bark to wood. The 6.5 mm lens of the Ken-a- Vision microscope slide projector was used to project the image of the microscope sections. A sketch of the section was drawn. The same method of determining the section center was employed as in the fiber ring study. Three radaii were drawn to the edge of the section. Keasurements of the bark and wood were along each radius. These measurements were used to determine the average thickness of wood and bark 'u (D H 1') (D O (“1" ion. Twenty sections per clone, and of each sex of T. cuspidata expansa were evaluated in this manner. b The means oi oark and wood per clone were determined. The two means then were used to determine the percent of bark to wood. ‘1‘ I -(' SULTS LLJ BEBE? KENT T - Propagation Results _— he rooting indices (Table 3) for cuttings of male versus female plants of Taxus cuspidata expansa indicate that cuttings taken from male plants rooted much better than cuttings taken from female plants. The ratio between the rooting indices was in the order of two to one in favor of the male cuttings. The difference between the means was found to be significant at the 13 level. Table 3. The Booting Indices of Hale vs. Female Cuttings of Taxus cuspidata expansa -'—' -F—nh-v —A— - .- deplications Female Kale Ratio 1 32 72 2.25 2 54 61 1.13 94 as 2.75 b 21 63 3.00 5 M2 90 1.90 6 3Q 7% 2.05 ? 33 70 2.12 19 77 4.25 9 27 32 1.19 1_ 25 .23 2.16 mean 31.4 *65.3 2.08 *significantly different at the .01 level 21 EIEEEIIETT II - Propagation lesults Differences in rootability were found in the seven clones of Taxus that were tested in this experiment. The average rooting indices and separation of the clones were determined by Duncan's multiple range test is shown in (Table 4). Taxus media 'datfield' proved to ca the s.perior rooting clone, while 1. cuspidata 'Tana' proved to be the —-—— —_ poorest in the clone experiment rootability. A comparison between the male and female clones revealed that the root- ability of female clones was superior. The difference was found to be significant at the 55 level. Table 4. gootin? Indexes of the Taxus clones ‘c-n ‘1‘3‘11'1 "fi'r -‘—'.“.J"" -v _, *-- -__V _ -n .. ' v'IHLD -1 '1. "r516: /}1fl *. A... ‘xAL/*_g.v_._-s .- +9 . 'u 'T“ w. "* 1'1 A- ' '11- “1“ ban a. . -fi-r..._.‘_.\ .a u_- - _ V ~ 71 M ' “‘ ' T" ).A‘hl d/a _. . 9A.; ._ :1 #v .4 fl 3‘ ' ‘.fi. *4 r- - 3‘ r' 3‘11 : fi1n 4.. .L. 4V;J.g_,‘.'. 4..~.L...4 ..--_'._./ .N__V ”TT “1‘ '“‘~‘—\~vq' T‘?'-,"' :3 “an ._ g -n . _J._ v 01“ A-.-»-'-_‘_'~.J haw v“ n r v— at o W.M01e l°fi 4.. a“. .......\---.-u_-. _. V..¢\/¢_...U [‘1 —--1- v at "q ‘ ’1‘ .fl‘\ -,1- A:. .f . .J. . |~v~’ Lj- U L} 1'~.Ll—4- U 9‘ .. . IVA— . 3v a'.§; __L \3 t 4‘ (\f‘ 3’? “J A 0‘11 .JV‘ 1,... s A---_t\‘.av. _. ‘ n.4- v A 11)“ '9‘-.‘-A- C 91 fi \‘ld v.-. H- u ,fi '3 ~._,‘ /. Lu ‘9 111;.) -v ‘1 M. 1* --\.~ L “L“Lb .J a O -- W 7‘37?“ '- lvaV s4~.»--- ’4 .t.‘.'$“" in. A4. L1,. V; -5 ‘."\"' _—~.’\v A h {ya-_‘l- -L H ..z -A ' \ -, , w‘agLJ -Li- “it-11* '1 '- ...J-.J — * I_I 1._ A... Is. ,1 VIA. 1‘r7‘1, 4' .1. L) '—’ _L C A1 .J H./ 3 in. J 1' ,v-« ‘ V..- V fl 1 V clo " . -AUV-n -.—~ 11 7- ‘_ .L ‘_. O 4..Av ”1‘- e b .l 0/ _\ 4... ‘k"; UZA :7 I "1 A VVAL Vr‘ -a'l Mh'lz m ,wi :- 1n s -43. ‘3 L‘ L1- (fin v- ”c. -,.‘1 ‘L - ‘.\J \J -‘L Us.) 0718 showed s ma 1..“ - '. -‘ . \ .ov-u‘ '1 r-F‘ " '1 / "'\._ .L 9‘ \ fl, . “q -51. l- ,A ,\ - 1 _A_ 'l ‘ _ ‘- A ' , ‘_ . v b. .l ‘_/ V v. ..... --- —“"— - _._ ..., _- ~.{ \. . - - ‘ _— v —r A a ' .. 1 '1 «m1 '1 .1. - 4" "J. kar- -- A ‘ ”vi-\V.o"; f‘, A '7'” Mr“ a. .A A ‘.V we —~/ 4 -- v. _ — .. Ir} 1 1 )1 I Q l _1 .1- H) l4. (LI .. t'\ \ (D J: / V «'3‘.‘ A ' “T "j 1 A‘fl..," ' r, Q ~. "\ Ouv tA- ‘- 4—waméd V'.~ ‘44,...3 J . / ’l .- -1— - v-- - ~ v z _ :1 ‘1 . \ '- _ v .. ‘ C -L girl __ Vac). w I. -.\..<.--..« J . \ L; J. .) ‘ C 3 .l J LI - 7\ ) ~ 0 r") P) " n ... «.1: s. I ,-.,, "i n -.-“‘|-1 m! . . _A— 4... ',v -4 -S .- Uo—b_1 .. _~ “A... he ‘ . \ 0 -~'- 1, 7'7 ".1 ’7 r1) n—dia '“lCHC' ,.5 ,., “'1— ,JV '2‘. \4 ”‘7' .4 J (‘C' " x.) ._. .- CU'V" V 4‘ Y} .i. - - ~.; F. A]: .,‘-) L‘-\ “TN/‘1? H“~‘f\‘? ~11. - -.—~ 4:1 4- .. .4'« 1.. -. .1. - .-4— .on, V -ne results or tLe studies enOn definite diiferences The reason for th superior rooting performance in nale 11 . . .sJ—r A - a} .0 . _ m cutt ngs of i. cuspidasa expensa over cne resale cuttin»s _ ‘40 97 *0 to w ' ‘i (D L3 ('1' O *0 *0 O t] H C t' H. O L C—t- 0 c—f C? (D ‘8 'D (i H '—-l O H h L) O ('l' S0 J I’J. H H. (‘1' Q J ., O ’_‘) female cuttings in the clonal study has not as yet been determined. It is *ossible that if an entirely different set of clones had been selected from the population, that he results might have been quite differen from those observed. f the survey U) U The data would seem to explain the result conducted by Eavidson (9) of Hichig an nurseries and land- I. scaee plantings, in which the ratio of male to female T. cuspidata expansa was found to be three to one. Plant 1 ) electing for male ( (:3 ators, unknowing_’ may Lth b~en um'v Taxus cu9iidata expanse due to the poorer roouacility of female cuttinr" Generally, nurseries select their cutting DUO i wood from the trimmings from plants wlich lave bee n line out in the field. Several decades of this practice could result in a preponderance of male over female Taxus cuspidata expan3a. Elgerton (11), Neal, et.al. (21) and Cliadwick (6) have suggested that the differences in root in; observed between the sexes of maple and nolly was related to the carbohydrate content of the cuttings. It was siggestei that since the female pla‘its produce fruit, they would have a lorer carbohydrate con tent;11ence cuttings taken from female ’3 '7 ».- ‘ plants would exhibit poore. :roota‘:ilitJ. If it were only a matter of carbohydrate supply, one would expect that all female clones would show lower rootability than the male . A 1 _.. ones. as .393 been not— 0 H (D Li: *1 1 1,. , . 4.1 , (sable s) his w s not the case; the male cl one of_ T. cusnilata 'Yana' had a low rooting ’3 "'V Y,““. . ‘ 4-, 1“! ‘rA,- V n .0 m - . ‘ ‘Ffl _— 1 .V \ ,- iniex, hulle tn- renale clones Ci 1. media '“lllozan' and ‘ ,1 . (' ' v v. . ' 1;. 1A [v.1 ‘— . y. ’ ('V, ". A / . mezid Nari had high rooting indices. snox (so) also noted female clones of maple wnich had high rootability 1‘ This would seem to indicate that another lac :34 tor in a Hui ion to ctloohjira content of the wood could be influencing the rootability f Taxus clones. r a t hormones and inhibitors could be a controlling p , A -\ L1 . ‘- v- P - ,-‘ t. _—\ - ‘ . n \ lactor in tne rootability 01 male 'ersus iema e .. CVSDluevfi expanse. Deboer (10) su$;€3ted that plant hormones and or the poor rooting of lnododendron s bearing flower buds. Whether the differences ors, or due to the high respiration rate of flower bud and the resultin3 low food supply COLCeDt as expressed a? Chadwick (6), or due to a comLination of several such factors must be decided he r experiment 1 investigation. 0" (.4 P. H ("r 0 Since the numoer of fibers found in the phloem of the sto ks showed a highIw bar: to wood ratio than the more vigorous rootstoc ks. From results observed in this exaeriment, there 3 G) O ers t be no relstionship betge en the characteristics 1 ‘ ~.. ~. 33 fins 1 .4 4-3 a o :1 hicn Jere oose rved DJ oealbane an- tLe ease oi rootin3 cl 5'71 ~ m. 1 1'1 3 .0- -y ,3 ,g 1 1.1.. . .. 4. -azus clones. -axus media ';atiieii' with the hi3hest “ .1. root n3 index also was found to have t '7' 4 ' Trww'fi ‘ ~. ‘I xfi‘: . - P' r ‘ . V‘: " f‘ “. . ‘1‘ good :atio (la le 7). Taxis cusfiidxta 'Idna' a slow growin3 ’3 ”no b ‘V - - v r' 17' ‘. ' semi-lust; iorm, use the lowes rootin3 inde“ of the clones able h); but only had a ledium thickness of bark to wood. *3 17- J 1 J.— :- 1. . - .o 1. 1.. 1 _- grog the results 0; the anatomica Tempures whicn Lere .. -. 4.. n ‘ ,3 4—1 L. 4.1. A 4.. . -- :- 31 _. 4. o . —.~~ 1 clonined, it seems that tne snag 3, ol ;arus cuttin33 has _fl - n . . . , J— ,... 1 _ L o J— . 11 4.1 . little in-luence in detorvinin- tne fOOQQbilluJ Oi the male versus female T. Olsbld ta expanse or of the dilferen. O J. l a J 2.6. 218' . 01238 O D J. 4 4.4.»)... q L1 C“ L.) .le ....;‘ r", \A Cf“. V Jr . f‘ ‘3 .7" ,4 I“ l l \,\p.. )- ME 1 “'3 I" 1:) _L...“v ‘~_/ i _I J. male «011 IN 1:, 1 4. and (_W A.) clone ,1 1 -L - 1 \ -,._.\,., .34. a”: m,—: v—fij a -d 131 ." ,._. H" ,"V ,. .L. '4 (:1 UV4-\- .t‘ lec l A. an "6' www.mmr A ., O ( T f S e C .l {L up d L 1 . mctnod and 11 3. 4. 3-”? p O f‘ .n- (w l w -. l .. 1. L)... V 4-1,. 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