SOME EFFECTS 0F COLD TEMPERATURE ON SEEDSTALK PRODUCTEON IN CARROTS Thesis éov ”19 Degree of M. S. MICHIGAN STATE UNIVERSITY Michaei Hugh Dickson 1956 SOME EFFECTS OF GOLD TEMPERATURE ON SEEDSTALK PRODUCTION IN CARROTS By MICHAEL HUGH ELCKSON AN ABSTRACT m . Submitted to the School of Graduate Studies of Michigan State University of Agriculture and Applied Science in partial fulfilhment of the requirements for the degree of MASTER OF SCIENCE Department of Horticulture Approved % ABSTRACT M. H. DIGKSON SOME EFFECTS OF GOLD TEMPERATURE ON SEEDSTALK PRODUCTION IN CARROTS The effects of cold storage on seedstalk induction in mature carrot roots and on seedling carrots were studied. Mature roots were harvested from Muck and Loam soils, stored for periods of l, 2, 4 and 8 weeks at temperatures from 32-500 F. The stored roots were subse- quently grown in the greenhouse at temperatures of 55-600 F. and 65— 700 F. The mature carrot roots when stored for 4 or 8 weeks at temper— atures between 32 and 50° F. and then grown in the greenhouse at 55- 60° F. produced 904100 percent seedstalks. Roots grown at 65-700 F. after eight weeks of cold storage also produced 90-100 percent seed~ stalks, but three months later than those grown in the greenhouse at 55-600 F. Those stored four weeks and then grown at 65-700 F. pro- duced between 33 and 90 percent seedstalks and they were likewise later than those grown in the cool greenhouse after four weeks of storage at 32-500 F. Roots harvested on August 30 and stored for eight weeks and then grown in the warm house produced fewer bolters and the seedstalks developed more slowly than those harvested October 3. Mature roots harvested October 3 produced seed earliest and in the most reliable manner when subjected to 8 weeks of storage at 40° F. followed by growing the plants at a greenhouse night tempera- ture of 55-600 F. until the seedstalks began to elongate. After the seedstalkaecame apparent the temperature should be raised to 70° F. to obtain rapid growth and flower development. ABSTRACT M. H. DICKSON Seedling carrots between 0 and 56 days old were not induced to bolt when subject to 28 days at 40° F. and then grown at 60-700 F. SOME EFFECTS OF COLD TEMPERATURE 0N SEEDSTALK PRODUCTION IN CARROTS By MICHAEL HUGH DICKSON A THESIS Submitted to the School of Graduate Studies of Michigan State university of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Horticulture 1956 ACKNOWLEDGEMENTS The author wishes to eXpress his sincere appreciation to Dr. C. E. Peterson for his assistance and supervision in planning the experiment, and in the preparation of the manuscript. The writer also thanks Drs. R. L. Carolus, S. H. Wittwer, and G. B. Wilson for their guidance in the preparation of the manuscript. Also to Dr. W. D. Baten for his advice on the statis- tical analysis. TABLE OF CONTENTS INTRODUCTION . . . . . . . . . . . . . . . . . . . REVIEW OF LITERATURE . . . . . . . . . . . . . . . EXPERIMENTAL . . . . . . . . . . . . . . . . . . . General . . . . . . . . . . . . . . . . . . Methods and Procedure . . . . . . . . . . . (l) The Effect of Cold Temperature on Flowerstalk Induction in Mature Carrots. (2) The Effect of Cold Temperature and Photoperiod on Seedling Carrots. . . . . Re sul ts . C C C . O O . . . C . . . O O C O (1) 0f Cold Temperature on Mmture Carrots................ (2) Of Gold Temperature and Photoperiod on Seedling Carrots . . . . . . . . . . DISCUSSION AND CONCLUSION. C O O O O O O O O O O 0 Some Effects of Cold Temperature on Seed— stalk DeveloPment in Mature Carrots . . . . Some Effects of Cold Temperature and Photoperiod on Seedstalk Development in Young Carrots . . . . . . . . . . . . . . . SUMMARY AND RECOMMENDATION . . . . . . . . . . . . L I TERATUR-E CI TED O O O O O O O O O O I O O O O O O 12 12 12 12 14 15 15 21 26 26 3O 33 357 INTRODUCTION The recent discovery of male sterility in carrots has en- couraged a plant breeding program for the development of male sterile lines, similar to those available in onions. Carrots being very heterozygous vary widely in shape, internal structure and composi- tion, even within a particular strain of one variety. With an open pollinated crop using mass selection improvement is very slow. Also to cross pollinate carrot plants by hand is very difficult and tedi- ous as the small flowers only produce two seeds. When male sterile plants were found a new opportunity for exacting carrot breeding in contrast to mass selection became available. With our present knowledge completion of the life cycle with— in a year using the greenhouse in the winter is a somewhat unreliable process. It was felt that further investigation into the flowering habit of carrots was required with a view to obtaining a reliable procedure for seed production during the winter months. Although the accepted method of inducing flowering in carrots is by a period of cold exposure, the minimum length of cold induction necessary for 100 percent seedstalk production in the shortest time has not been found. In order to study the optimum temperature for induction, and the necessary minimum thermoperiod an experiment was designed to find a reliable method of post harvest induction to insure rapid seed production. In most commercial plantings there is a small percentage of early bolters in various stages of development which must be rogued before harvest to insure a high quality processed product. Steckling stocks from which early seeders have been removed still contain roots with a tendency to bolt and are the source of commer- cial seed. An investigation of conditions under which early initia- tion occurred was necessary. Therefore, an experiment was designed to see if any carrot seedling induction would occur under normal conditions and to determine the effect of day length and temperature of growth after cold induction process. If a reliable test could be found for promoting induction of susceptible carrots in the first season more adequate rogueing could be made with eventual elimination of the early bolters. At the same time a means to induce premature seeding would be useful in a plant breeding program to hasten back crossing and testing for male sterility. REVIEW OF LITERATURE Effects of cold on flower induction in various plants. Gassner (8) was among the first to investigate the effects of cold treatment on flowering and seed formation. He observed that beets held in a cool greenhouse from January to April went to seed the following Summer, while those held at 20° 0. did not. Also that cabbage, rutahaga and carrots seemed to depend generally for their flower formation upon the influence of low temperatures. Boswell (2) reported that in onions flower primordia differ- entiation was almost entirely inhibited by storing for 8 months at 320 F., while storage at 50° F. produced a much higher percentage of flowers. However, 6fi-months at 320 F. followed by 7 weeks at 50° F. greatly increased the flowering percentage compared with bulbs stored 8 months at 32° F. He also observed that the lower the storage tem- perature below 50° F. the greater the inhibiting effect on flower primordia development and that the large bulbs produced higher per- centages of seeders. Thompson and Smith (29) working with onion sets state that large sets of 13/16-1 1/8 inch diameter produced much higher per- centages of seedstalks than did medium 5/8-3/4 inch diameter or small sets 3/8-5/8 in diameter. They observed that sets stored at 40-500 F. produced the highest percentages of seedstalks as well as the fewest marketable bulbs. Sets stored at 320 F. produced the best bulbs, and while sets stored at 60-700 F. also produced few flower stalks the bulbs were much shriveled. When sets were stored under conditions unfavorable for seedstalk production large sets produced the best yields, but also the most splits and doubles. For seed production a growing temperature of 50-600 F. was found to be best, while at 70- 800 F. seedstalks did not develop. When sets in storage at 320 F. were moved to 40-500 Fh,seedstalk production increased compared to those held at 320 F. all the time. Moving sets in storage from 40-500 to 60-700 F. decreased seedstalk production and moving them from 60-700 F. to 500 F. or 32° F. increased seeding. Increas- ing the photoperiod by five hours over the normal winter day length hastened seedstalk development, but under high temperatures of 70- 800 F. seedstalks did not develop under normal day length or under long days. Jones (11) and Jones and Emsweller (12) working with the Ebenezer variety of onion found that bulbs stored at 46° F. and 530 F. always produced the earliest seedstalks while those stored at 860 F. produced the latest, and also that high temperature produced vegeta- tive bulbs and few seedstalks. Bulbs stored at 53° F. always produced the largest seed yields. Starring (25) in.flontana working on celery, found that a check in growth due to cool weather alone unfailingly caused seed produc- tion. He also found that at longer periods of cold temperature the percentage of bolters increased. Thompson (26) stated that a serious check in growth delays seedstalk develOpment in celery and may prevent it entirely. In young seedlings after 2 weeks or more at 40-500 F. lxnmature seeding is likely, also plants grown in the greenhouse for 2é-months or more at 500 F. are likely to produce seedstalks in the first season. However, a 70° F. temperature after 30 days of cold prevented seeding unless the seedstalk was started before subjection to this temperature. For the best seed production he recommended growing the celery at 55-6o° F. after the cold induction until the seedstalks had started to grow, then completing growth at 60-700 F. He also stated that when conditions are favorable for rapid growth for a few weeks following the cold treatment, the percentage of seed- stalks are much higher than when conditions are unfavorable for good growth. This applied to beets as well as celery. Pawar and Thompson (15) investigating the effects of age and size of plant at time of exposure to low temperature,observed that older celery plants at the time of cold treatment went to seed faster, but that the total time from seed to seed was shortest when 2 month old plants were treated. However, plants treated at any age would bolt eventually. The age of the plant was the important factor, and size had no effect on the rate of flowering. In studies on premature flower formation in wintered-over cabbage, Boswell (3) found that below a stem diameter of 6mm the increases in seeding with increasing size of plant are small. At a diameter of 6-7mm there is a sharp increase in the percentage of seeders, above which the increase is still more marked. Earlier planting in the fall produced increased bolting even if the over- wintered plants were the same size. This appeared to be due to the older plants being physiologically more mature. Low temperature is effective in promoting reproduction only after the plants have accu- mulated a sufficient weight of reserve foods. .fliller (13) found that cabbage plants bolted a month earlier when grown at 60-7o° F. than at 50-6o° r. after 2 months at 40° r., but that 15 and 30 days at 400 F. only produced a very few bolters. He also found that increasing the day length did not enhance seed- stalk develoPment. Chroboezek (6) observed that garden beets became vegetative at high temperatures even after the flower stem had grown to full height and the buds had developed. It was shown that the crown or growing point was the vital area, which had to be given cold treat- ment to induce bolting. He was able to produce bolting by winding a small rubber tube around the base of the petioles and circulating cold water at 43.80 F. If the tubing was wound around the base of the root no seedstalk production occurred. He also noted that germination at low temperature did not induce flowering in beets which were later grown at high temperatures. Thirty days at 40-500 F. for young plants induced flowering when grown later at 6o-7o° F., but 7o-80° F. nul- lified the cold treatment. However, if the plants were grown for 60—90 days at 40—500 F. the nullifying effect of high temperature was not evident. Extra light helped seed production at high temper- atures but under cool conditions 8 hours was enough. However, at high temperatures very few plants went to seed even with extra light. The most plants bolted when they were grown at 50-600 F. and under 15 hours light. Plants grown at 50-600 F. under continuous light produced seed stalks 53 days after planting. Naylor (14) stated that by using continuous light of 1000- 1400 ft. candles flowers were produced in 31 days in garden beets and 40 days after seeding in dill. At 500 ft. candles seed stem elongation in 60 days was observed in dill, but none with this light in biennial hosts. The intensity of the light proportionately speeded bolting in dill. Cold suppressed the flowering responses under con- tinuous light. Annual beets would not flower if more than 10-11 hours of darkness were given in 24 hours, but 13 days of continuous light resulted in bolting even on returning to short days. Annual beets required over 700 ft. candles for flowering. In studies on premature seeding in beets Smith (23) found that when mature beets were grown in the dark at 550 F. no flower primordia were formed, but if in the light 100 percent flowered. However, two weeks in the dark at 50° F. followed by continuous light at 55° F., produced more bolting than growth in continuous light at 550 F. Plants grown at 500 F. for 2 weeks and then moved to a grow- ing temperature of 550 F. and continuous light produced the most seedstalks. Thirty-seven to forty-seven day old plants responded better than older or younger plants. Carolus (4) stored beets and turnips at 32-400 F. and also 55-650 F. in common storage. The cold storage temperarily delayed seedstalk elongation compared to common storage; however, the re- sultant flower stalks were longer and more vigorous from the cold stored plants. For each month of storage the rate of seedstalk elongation was increased on removing the plants from storage. Thus, beets stored 3 months reached 90% elongation ahead of those stored 2 or 1 months, although planted l and 2 months earlier, and all reached 100% at the same time. While in turnips, 2 weeks after planting,plants stored 3 months were ahead of those stored 2 months, although the latter had been planted 6 weeks. Peto (16) reported flowering in turnips was inhibited by temperatures above 65° F. in plants which had shown incipient bolting, and that plants grown continuously at high temperatures produced vegetative growth, while low temperatures favor sexual reproduction. Sakr (21) reported that when turnip plants one month old were subjected to 30 days at 40-500 F. subsequent exposure to 50-600 F. was more favorable for seedstalk production than 60-700 F. or 70-800 F. After 60 days of cold storage at 40-500 F. any growing temperature gave a high percentage of bolters. He found that increased light in Nevember and December speeded seed production at higher growing temperatures. He also found that seed vernalized for one month at 370 F. produced 64 percent bolters if it had germinated during vernalization, but only 3 percent if it had not germinated. Plants put into the cold when just germinated tended to bolt more than those put into the cold when one month old. A temperature of 530 F. and continuous light on germinating seedlings gave incipient bolting in 71 days. In mature plants 30 days at 40-500 F. followed by growing at 50-600 F. gave the most seeders. If the temperature following exposure to 40-500 F. for 30 days was 650 F. or above bolting was inhibited to some extent. ' Sakr (22) reported that no bolting occurred in seedling parsnip plants grown 33 days at 60-700 F. followed by 15, 30 or 60 days at 40-500 F. and then returned to a temperature of 60-700 F. However, 100 percent bolting resulted when 4 month old plants were subjected to 35 or 49 days in cold storage at 40-500 F. and subse- quently grown at 60—700 F. Chesnokov (5) stated that in turnips, cabbage and carrots young seedlings,rather than slightly germinated seed, chilled 50 days gave a higher percentage of seedstalks. He found with best plants that 80-90 percent bolted the first Summer when young seedlings were chilled 50 days. Verkerk (30) reported that the first signs of the generative stage were visible in disected growing tips of brussels sprouts after 4 weeks at 5 or 7° C. Vlates and Meudt (31) in investigations on vernalization and photoperiodism in Spinach observed that seed vernalized at 20 C. for two weeks produced up to 25 percent bolters at any photoPeriod above 8 hours. After 8 weeks of vernalization at 20 C. all the plants flowb ered in 35 days when grown under 12 hours photoperiod as compared with only 8 percent flowering among plants grown from seed held at 250 C. It was shown by Thompson and Knott (28) that high temperatures also have a role in flower induction. Lettuce grown at 70-800 F. went to seed without forming heads, while similar plants grown at 60-700 F. formed heads and then went to seed. Cochran (7) working with peppers found that blossom buds developed at 60-100O Fh,but at 90-100o F. they dropped off without setting fruit. Pepper plants at 550 F. fail to develop flower buds, but if plants with buds were grown at 550 F. all developed fruit parthenocarpically. In studies on vernalization Gregory and Purvis (10) and Purvis and Gregory (1?, 18) observed that a short period of high temperatures (300 C.) devernalized rye grain, but that it could be revernalized in a shorter time than the original vernalization. The degree of devernalization varied inversely with the duration of the previous vernalization. Three days at 35° C. after 4 weeks vernali- zation at l0 C. resulted in 55 percent devernalization. After 6 weeks 10 only 16 percent devernalization occurred and after 12 weeks none occurred. Also, the vernalized condition was stabilized against re- versal if a period of growth at 15° C. immediately preceded the high temperature. With mature carrots Sakr and Thompson (19) found that storage at 400 F. resulted in a higher percentage of bolters than storage at 35° F. or 500 F. Roots stored 15 days at 400 F. and then grown in the greenhouse at 50-600 F. produced 100 percent bolters, while 30 days storage at 35° F. and 60 days at 500 F. were required to produce 100 percent bolting at the 50-600 F. growing temperature. Storage for 60 days at 400 F. was required to produce 100 percent bolting when plants were grown in the greenhouse at 60-700 F. He also found a slight advantage in using natural day length of about 10 hours during the winter rather than continuous light. The check plants grown con- tinuously at 50-600 F. without a previous cold treatment produced as many bolters as those stored 30 days at 500 F. and then grown at 50-600 F. He decided that a growing temperature of 50-600 F. was the most favorable for seedstalk development, except after a long period of 80 days or more at 400 F. when even at the higher tempera- tures all plants went to seed. Carrot seedlings (Sakr 20) subjected to one month at 40-500 F. starting 17 days after planting produced more bolters at a growing temperature of 50-600 F. than when grown at 60-700 F. and 70-800 F., but in all treatments the percentages were low. When seedling plants were grown for 135 days at 40-500 F. then moved to 50-600 F., 54 percent bolted, while those grown at 60-700 F. following the same treatment produced 43 percent bolters. Light increased vegetative 11 growth at low temperatures, but made less difference at high tempera- tures; it made no difference on seedstalk production. 12 EXPERIMENTAL General Flower induction was investigated for two types of carrots. One involved mature carrots harvested in the fall, stored for vary~ ing periods at different temperatures and then grown in the green- house. The other involved seedlings of different ages subjected to a cold treatment and then grown at different temperatures and photo- periods. Materials and Methods of Procedure The Effect of Cold Storage on Flower Stalk Induction in lature Carrots Qfiggots ggown at the Nuck_§ggg. Carrots of the variety Long Ghantenay were seeded at the Experimental Muck Farm on May 3, 1955. Nature roots were harvested on August 30 and selected for trueness to type. .The tops were out off about one-half inch above the crown and the roots were then dipped for approximately 30 seconds in an antibiotic solution*. A total of 600 roots were selected and placed in shallow wire- bottomed trays in moist sphagnum moss, l20 roots being stored at each of five temperatures, 32, 40, 45, 500 F. and the check lot at room temperature, approximately 700 F. * Agri-mycin 100, formulated by the Pfizer Company, was used at a concentration of 150 ppm as recommended. It contained Streptomycin 15% and oxytetracycline 1.5% as the active ingredients. 13 Storage periods of l, 2, 4, and 8 weeks were used. At the end of each period 20 carrot roots were removed from the trays in each temperature and planted in the greenhouse in 6 inch pots. Thus, the plants with the longer storage periods were planted later. For the ’ first planting the soil was rather heavy and there was about 50 per- cent loss during the following month mostly due to bacterial soft rot. The second and all the following plantings were grown in a mixture of equal parts of loam, sand and muck. The third lot was planted in a core of Arasan and sand in an effort to prevent rotting, but after a period of two weeks this was found to inhibit root development so they were repotted in the soil mixture without a sand-Arasan core. Carrots for the fourth planting were rolled in Arasan or Spergon and then potted. Until November 25 the greenhouse night temperature was 700 F. and often reached 900 F. by day. From November 25 to the end of the experiment it was possible to hold the greenhouse temperature near 65° F. at night and 700 F. by day. Due to an accident the tempera- ture reached 1000 F. during the night of November 21. Carrotsggrown at the Horticulture EEEE: These carrots were seeded on a sandy loam outlay 11, 1955, at the Michigan State Univer- sity Horticulture Farm. On October 3 mature roots were hand harvested. The tops were cut off one inch above the crown, as it was felt that slightly longer teps might better protect the apical bud. They were dipped in the same anti-biotic solution as roots harvested at the Ifluck Farm, then placed in moist sphagnum moss in shallow wire-bottomed trays and kept in storage for periods of l, 2, 4, and 8 weeks at 32, 40, 45, 50° F. The check lots were stored at a temperature of about 70° F. for corresponding periods of time. 14 On being taken out of storage the roots were rolled in Arasan or Spergon and then potted in the loam, sand and muck mixture. They were then divided into two lots, one lot to be grown at a night tem- perature of 550 F. and the other at 650 F. It was not possible to achieve the 650 F. intended until November 25, prior to that date the temperature was 700 F. The cool house was not available until October 18 so the plants stored for 1 week were placed outside in the cold frame. As a result they received some further cold treatment since the outdoor temperature averaged a little below 500 F. Until the end of October, the cool house had a night temperature of 600 F. after which it was fairly constant at 550 F. except for November 24-29 when it was at 65° F. During the day the cool house was at 55-600 F., ex- cept on 7 bright days in February when it reached 700 F. No check was used with the roots stored 4 or 8 weeks as the crowns were eaten by mice during the storage period. Primordia as indicated in Tables II and III were based on a visual inspection made by cutting the carrot through the crown on March 3 when the experiment was terminated. Figure I illustrates the stages observed in the de- velopment of Primordia. The Effect of Cold Induction on Seedling Carrots Seedling Carrots Growing in Greenhouse Carrots of the variety Red Cored Chantenay were seeded on September 23, October 7, October 21, November 4, November 18 and De- cember 12, l955,direct1y into 2% inch pots in a soil composed of equal parts of loam, sand and muck. Thirty-six pots were seeded on each date. On November 18 the seedling plants were all placed in cold 15 storage at 400 F. until December 16. The seedlings had roots of about .5 to .7 centimeters in diameter. The check was seeded on December 12 and received no cold storage. On being taken out of storage the pots were divided into two groups, one being put in a greenhouse at a night temperature of 650 F. and the other at a night temperature of 600 F. The temperature of the warm house during the day was usually around 75-800 F., while the cool house seldom exceeded 650 F. The plants were repotted in 4—inch pots when their size required it. In each house there were three further divisions, with one lot receiving 8-9 hours, the second 12-13 hours and the third lot 16-17 hours of light in each period of 24 hours. The natural light from December 16 to February was very dull and this may have further retarded growth of the 8 hours lot which had no supplementary light. Results Carrots grown at the Mbck_§a£g, The plants which had had 8 weeks of storage irrespective of temperature (see Table I) had pro- duced about 34% bolters by January 19. However, for all practical purposes the maximum storage period of 8 weeks was inadequate to pro- duce rapid belting under the subsequent growing conditions of an average temperature of 700 F. Roots which had been stored for l or 2 weeks were discarded in early December when it appeared that they had become senescent and were not bolting. Roots stored for 4 or 8 weeks were discarded on January 19. Those plants which had bolted by October 29 must have been pre-induced in the field as they bolted almost immediately on being taken out of storage. Data for the second Figure I. Carrots showing development of seedstalk pri- ncrdie. Top row left no princrdie develomentg bottom row right, advanced princrdie. 16 17 group of carrots harVested at the Horticulture Farm October 3, are presented in Tables II and III. These tables list the number of plants showing seedstalk elongation by each date to March 3, and the number showing some degree of flower primordia formation on March 3, when the experiment was terminated. The most outstanding difference in bolting was that which was due to growing temperatures after the roots were taken out of storage. In every case the carrots grown at 55-600 F. after being taken out of cold storage bolted earlier and in a higher percentage than those grown at 65—700 F. One week of cold storage was insufficient to induce bolting and the plants were discarded on January 21 (Table II and III). Two weeks in the cold storage induced an averag6260% bolting by March 3 in the cooler house and 35% in the warm house, if plants showing pri- mordia are also included in the totals, but only 46% and 12% when only plants with visible seedstalks were included. Four weeks cold storage resulted in a high percentage of bolting in both houses. When all plants showing any degree of flower stalk develOpment were in- cluded the bolting percentage was 95% in the 55-600 F. house and 67% in the 65-700 F. house. On the basis of visible seedstalks only, there were 85% in the cool house and only 36% in the warm house. In both houses the plants given 8 weeks of cold storage pro- duced almost 100% bolting if those showing flower primordia were included. 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