\ WHMINMIHM H h WW fl WWI I—4|\)_3 ‘ row lln .m-P-OO R t I!!!lllllllllzlfljfllllllflllflwlflflll!Nfllllfllfll LIBRARY 7 Michigan Stat: ’ , University . This is to certify that the thesis entitled YIELD OF CROPS TREATED WITH 1—TRIACONTANOL presented by Terry Lee Richman has been accepted towards fulfillment of the requirements for Masters . Horticulture degree 1n Major/ rofessor Date 2’/2’ é/7 7 OVERDUE nuns ARE 25¢ PER DAY . PER man Return to book drop to remove a this checkout from your record. M8111? YIELD OF CROPS TREATED WITH 1—TRIACONTANOL By TERRY LEE RICHMAN A THESIS - Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Horticulture 1979 ABSTRACT YIELD OF CROPS TREATED WITH l-TRIACONTANOL BY Terry Lee Richman The growth and yield of several vegetable and field crops were sometimes increased by applications of l-triacontanol (TRIA) in 1977 and 1978. Increased yields from TRIA applications were obtained over a wide range of rates. A single application of TRIA was as effective as multiple applications. There was no apparent optimum growth stage for application. The response of carrot (Daucus carota) and sweet corn (Zea mays) to TRIA was greatest when grown under lower nitrogen regimes. Soaking seed of crOps in dichloromethane containing TRIA did not prove to be an effective method of application for crops directly seeded in the field. However, it was found effective for tomatoes (Lycopersicon esculentum) grown in the greenhouse and transplanted to the field. Granular formulations of TRIA, placed on the soil surface, increased the yield of wheat (Triticum aestivum), tomato and asparagus (Asparagus officinalis). The granular formulation of TRIA, placed with the seed, was found to decrease the yield of cucumbers (Cucumis sativus). ACKNOWLEDGMENTS I wish to thank Dr. Stanley Ries for his aid in my research and Drs. Alan Putnam, William Meggitt, Everett Everson and Hugh Price for their guidance and assistance. I would like to acknowledge the coopera- tion of the farmers and county agents of Michigan, particularly Gregory Varner and Edgar Strong, for their willingness to provide their assist- ance, without which many of these studies would not have been possible. I also wish to express my appreciation to David Glenn and other friends, for their support throughout these studies, particularly Dr. Jennifer Jones for her assistance in writing this thesis. ii TABLE OF CONTENTS Page LIST OF TABLES . . . . . . . . . . . . . . . . . . . . . . . . . . iv LIST OF FIGURES . . . . . . . . . . . . . . . . . . . . . . . . . . vi INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 LITERATURE REVIEW . . . . . . . . . . . . . . . . . . . . . . . . . 2 & MATERIALS AND METHODS . Formulations and Methods of Application . . . . Seed Treatments . . . . . . . . . . . . . . . . . . . . . . . Foliar Treatments . . . . . . . . . . . . . . . . . . . . . Soil Treatments . . . . . . . . . . . . . . . . . . . . . . Statistical Methods Ht—I mutants RESULTS AND DISCUSSION . . . . . . . . . . . . . . . . . . . . . . 20 Seed Treatments . . . . . . . . . . . . . . . . . . . . . . . 20 Foliar Treatments . . . . . . . . . . . . . . . . . . . . . . 23 Soil Treatments . . . . . . . . . . . . . . . . . . . . . . . 35 CONCLUSION . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 LITERATURE CITED . . . . . . . . . . . . . . . . . . . . . . . . . 42 iii Table 10. 11. 12. 13. 14. LIST OF TABLES Page Cultural conditions for crops soaked in DCM containing TRIA in East Lansing in 1977 . . . . . . . . . . . . . . . . 6 Cultural conditions for alfalfa-triacontanol comparison studies in East Lansing in 1977 . . . . . . . . . . . . . . . 7 Cultural conditions of 'Heinz 1350' tomato soaked in DCM containing TRIA in 1978 . . . . . . . . . . . . . . . . . . . 8 Cultural conditions for crops receiving feliar applications of TRIA in 1977 and 1978 in Central Michigan . . . . . . . . 10 Cultural conditions for crops receiving foliar sprays of TRIA at multiple intervals in 1977 . . . . . . . . . . . . . 12 Cultural conditions of wheat treated with foliage sprays in Central Michigan in 1978 . . . . . . . . . . . . . . . . . 14 Cultural conditions of tomato and dry bean crops receiving foliar sprays at various stages of growth in 1978 . . . . . . 15 Cultural conditions of carrot and sweet corn grown on different nitrogen levels . . . . . . . . . . . . . . . . . . 16 Cultural conditions of several crops receiving TRIA applica- tions to the soil around the plants in 1977 and 1978 . . . . 17 Cultural conditions of wheat receiving granular applications of TRIA in 1978 . . . . . . . . . . . . . . . . . . . . . . . 19 Yield of several vegetable crops when seed soaked in DCM- TRIA solution for one hour in 1977 . . . . . . . . . . . . . 21 Yield of several crops with seed treatments of alfalfa and DCM-TRIA soaking in 1977 . . . . . . . . . . . . . . . . . . 22 Response of 'Green Star' pickling cucumbers to TRIA applied as a granular formulation with the seed in 1978 . . . . . . . 26 Yield of several crops receiving foliar applications of TRIA in 1977 . . . . . . . . . . . . . . . . . . . . . . . . 27 iv Table 15. 16. 17. 18. 19. Page The yield of several crops receiving single or multiple applications of TRIA in 1977 . . . . . . . . . . . . . . . . 29 Response of wheat cultivars to TRIA applied in different formulations, times and rates at Saranac and East Lansing in 1978 O 0 O O O 0 O O O O O O O I O O O O O O O O O I O O O 31 Yield of cucumber, tomato and navy bean crops when treated with foliage sprays of TRIA in 1978 . . . . . . . . . . . . . 32 A comparison of yield of "snapped" 'Mary Washington' asparagus after treatment with TRIA at Sodus, Michigan in 1978 . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 Yield of 'Heinz 1350' direct seeded tomatoes as influenced by granular applications of TRIA . . . . . . . . . . . . . . 39 LIST OF FIGURES Figure Page 1. A comparison of 'Heinz 1350' tomato yield from TRIA treatments applied to the seed and to plants after flowering in the field . . . . . . . . . . . . . . . . . . . 25 Yield of 'Spartan Classic' carrots treated with TRIA at two different N levels . . . . . . . . . . . . . . . . . . 34 Yield of 'Gold Cup' sweet corn at different rates with different levels of N nutrition . . . . . . . . . . . . . . . 37 vi INTRODUCTION Triacontanol (TRIA), a constituent of leaf wax among plant species (5,6,7,12), has been shown to increase the dry weight of several crops (11). Greenhouse studies demonstrated that TRIA, applied to the seed, soil or foliage improved the growth of several crops (9). The objective of this research was to determine if TRIA could be developed as an effec- tive growth regulator for increasing crop yield. These field studies were initiated to determined the optimum rate, number of applications, optimum time and best method of applying TRIA to increase crop yield. LITERATURE REVIEW Coarsely chopped alfalfa (Medicago sativa) hay increased plant growth and yield when placed in a band below and to the side of crop seeds or seedlings (8). A crystalline substance, isolated from alfalfa meal, was identified as l-triacontanol [CH3(CH HZOH] by 2)28C mass spectrometry and was shown to increase the dry weight and water uptake of rice (Oryza sativa) seedlings grown in nutrient culture (11). It was also shown that foliar sprays increased the growth of barley (Hordeum vulgare), corn and tomato grown in soil. The response of rice and tomatoes to synthetic and natural triacontanol was similar. Triacontanol was first identified in 1933 as the principle long-chain alcohol component of wax derived from alfalfa leaves (2). TRIA occurs in different quantities in the wax of many plant species and is present throughout the environment (5,7,12). TRIA was found to be present in the inner tissue of several plant species suggesting a role aside from cuticular functions (6). TRIA increased the dry weight and leaf area of rice seedlings grown in nutrient culture at 2.3 x 10-8 M (10 ug/l) within 3 hours in the light or dark (10). TRIA also increased Kjeldahl- N content up to 30% after 6 hours in the dark. The dark response is sensitive to atmospheric CO2 and 02 concentration (1). Treated rice seedlings increased dry weight in the dark in the presence of 200 to 400 ppm atmospheric CO2 and 5% 02. The treated seedlings did not fix more atmospheric C02 in the dark. TRIA increased the protein content 2 3 and growth in_vitro of cell cultures of haploid tobacco (Nicotiana tabacum) (3). The increase in growth was due to an increase in cell numbers. Octocosanol [CH3(CH HZOH], the 28-C analog did not increase 2)26C the growth of tobacco cell cultures (3) or the growth of rice seedlings (10). Octocosanol inhibited the response to TRIA at 2.3 x 10"8 M in rice seedlings at concentrations as low as 2.4 x 10'12 M (4). Primary alcohols with carbon chain lengths of l6, 18, 22 and 24 also inhibited the TRIA response in rice seedlings indicating that chain length, presence and position of hydroxyl group may be specific for the growth promoting activity of TRIA. The growth of several vegetable and field crops in the greenhouse was increased by applications of TRIA to the foliage, soil or seed (9). The dry weight increase occurred at applica- tion rates as low as 0.01 mg/l and was maintained over a lOOO-fold range in concentration. Clearly, the efficacy of TRIA has been demonstrated as a compound capable of increasing the growth of many crops in greenhouse and labora- tory situations. There have been no reports of TRIA increasing yield of crops in the field previous to the initiation of this study. MATERIALS AND METHODS Formulations and Methods of Application TRIA was applied in the field studies as a seed treatment, foliage spray or a soil treatment. TRIA was applied using formulations supplied by the American Cyanamid Company, Princeton, New Jersey. Foliar appli- cations were prepared from a 2.5% or 0.01% concentrated emulsion. Due to the small amount of the emulsion required for applying the low rates of TRIA, Tween 20 at 0.1% by volume was utilized as a surfactant in all foliar studies. Controls consisted of Tween 20 or emulsion blank in all studies. All foliar treatments were applied with one or two flat fan nozzles at rates of 423 l/ha to 984 l/ha in 1977. In 1978, foliar treatments were applied with one flat fan nozzle or 2 even spray nozzles at rates from 225 l/ha to 438 l/ha. Seed treatments in both years con- sisted of soaking the seed in dichloromethane (DCM) containing TRIA (0.01 to 10 mg/l) for one hour. Controls consisted of seed soaked in DCM for 1 hour and untreated seed. The seeds were dried at room tempera- ture prior to planting. In studies involving granular applications of TRIA, fbrmulations of 2.5 x 10-4%, 1.0 x 10-3%, 4.0 x 10-3% and a granu- lar blank were utilized. Seed Treatment Seed of several crop species; carrot, sweet corn, cucumber, lettuce (Latuca sativa) and radish (Rhaphanus sativus) were soaked in DCM 5 containing TRIA (Table l). The cr0ps were planted in East Lansing using a V-belt planter. Crops were irrigated frequently to prevent wind erosion until established. Weeds were controlled by hand cultivation. In experiments comparing alfalfa hay and TRIA applications on the yield of barley, sweet corn and tomato, the seed were soaked in DCM prior to sowing or were sown with finely ground alfalfa hay (Table 2). Crops were planted on a Spinks sandy loam soil. All treatments received 1.0 g finely sifted vermiculite to hold the alfalfa hay on the planter. Weeds were controlled by hand cultivation. In 1978, 'Heinz 1350' tomato was the only crop studied using the method of soaking seed in DCM containing TRIA (Table 3). Transplants from control and treated seed were grown in plastic flats (26 x 53 cm) containing vermiculite. The flats were placed in a growth chamber with a day-night regime of 16 hours (25°C) and 8 hours (20°C). After 15 days, seedlings were transplanted into wooden flats (35.5 x 51 cm) containing a greenhouse soil composed of sand, peat and loam (1:1:1) and placed in a greenhouse with the night temperature maintained at approximately 27°C. The plants received 0.5 1 of a water soluble 20- 20-20 fertilizer twice a week (1 g/l). Forty-two day-old plants were sorted by size for blocks and placed in a growth chamber for the night (16°C). The plants were transplanted with a commercial transplanter. Additional TRIA treatments were applied as a foliage spray (50 and $00 mg/ha) or a granular topdressing (0.4, 1.6 and 6.4 kg/ha) in 12.7 cm bands on each side of the row. Granular formulations of TRIA were placed with the seed of 'Green Star' pickling cucumbers on a Spinks sandy loam soil in East Lansing in 1978. The crop was planted June 6 with a V-belt planter to a depth NH-~H-- a;\mx com w.m c.5xm.o oamwuass m\w mH\m NH\m seem flew“: NH-~H-NH m:\mx cow m.H c.5xo.o mamcflm o~\o mH\m oH\m Eon Ap:¢m mxcwmm NH-~H-~H mg\mx cos m.H o.nxo.o ofimcflm nH\o vH\v mH\v EmoH chmm mxcflmm NH-~H-NH m:\mx cos o.N c.5xc.o ofiwcflm 0\n 4H\4 mH\v EmoH xpcmm mxcfimm NH-~H-~H m;\mx cow N.m o.nxm.o mamfiuase om\n mH\o mH\o EmoH xccmm mxcflmm pofiaaam upon Agog canon may onwm pon umo>hmg we on%& camp umo>nmx pump mcwpcmfim camp acoEumone wasp snow .QSU UHOD. E00 ”003% .Huom Apnoea. amnemu .nocshououusm. monuuoa .ofimmmau cmupmmm. pounce Hm>wufisu was mono .howEoum. honESUSU .ahmfi an .maumamg pmmm :« waov m:\~ coon-H voowua voowug moomufi mafim ofiuuoz o.egm.o 0.5xo.o o.exo.o N.meoe.o flee «nun “one vanguasa ofimzfim ommcwm uncfim umo>umn mo omxb mH\w c~\m AH\o 4\m mane pmo>umz mama vnnuvcm mama cam mama vumlvcm mama can mono mo ommum oH\o ~H\m ecu m\m mm\m use wH\m wm\o camp pcoEumonb unon-> uflon-> puma-> Henge Hanuuoasoo eczema menueaga mN\m mu\q «H\v oH\o came «cupcaaa emoH awed emoH Emofi “an“: xvcmm mxcwmm xvcmm mxcwmm mafia fiflwnxumm waxy Hwom nnma nnma nnmfi mead new» .930 vfiou. .HHom xnuonu. .cucshonouuam. .nmum coonu. :uoo poozm nmflvmu ouduuofi sonssoau Hm>wuasu can mono .aamfiaonz Hmnpcou a“ mnmd was head :a fiooou macho pom m:0fluwv:ou Hmhzuasu .v oanmh 11 Am:\.w.m mm houmoaxcuoa ~.Hv ocflnwnum weaponuvcmn mzfimonnucm: consmuoHso Hahucoo poo: V\o :o m:\z we was mafia NHuNHnNH NH-~H-NH ~H-~H-NH NHuNHuNH an\mx com a=\mx oom a:\wx com mn\mx mmm eonuddm upon .msu vaou. .HHom xuhonu. .noqahououpzm. .uwum cacao. :Hou poozm :mwpmu ousupoa nonasuzo um>fluasu can mono fieoseupcouU q magma 12 has \.n.a we e.mv euaacagane --~H-~H m;\mx woos ecu cam mmv 0.5xm.u vanguase o~\w mcfinozoam umH oH\o m~\m 5.2 23: ueam ecu eufimusfimnau NH-~H-NH m:\mx mum MNV «.muxm.o oawcwm EN\m oumfifiomflup emu efi\o m\o amofi smHu Hflugxuma w:«oo::v:m: NH-~H-NH mn\wx mme mmv 0.5xm.o ofinflumse om\w mama paw nm\n m~\h Smog xucmm mxcwmm Enamumm:.v:m :onamuoHsu NH-NH-NH m:\mx wee mmv N.mflxu.o ofiwcflm wa mama van m~\o oH\o awed smug Hfiuexcaa H Oh HGOU U002 vofifimnw.uuom aE\H hey ouflm uoHd pmo>nms mo omxh camp umo>nmx mono mo ommum camp unusumouh oumv mafiucmHn omxu aflom .ommH Nader. oumaou .umaficmm. amen xnv .Hofisonm. Honasoso nm>wufiau can mono .umum :oouu. umpESUSQ .nnmfi :fl mHm>houcfl oamwpasa um fiooon mmouu pom mcofluwvaoo Hmnsuaso .m oases 13 Navy bean, tomato, and wheat plots were tested in 1978 in an attempt to determine the optimum morphological stage of growth with respect to TRIA applications (Tables 6 and 7). In the wheat and navy bean tests, two guard rows were placed between each treated plot to avoid spray drift. The wheat plots were harvested with a small plot combine. Nitrogen studies were conducted in both years in East Lansing on a Spinks sandy loam soil (Table 8). Soil Treatments Soil applications of TRIA consisted of water drenches and of granu- lar formulations applied around the plants (Tables 9 and 10). Statistical Methods Factorial experiments were utilized in most field studies with rate, number and morphological stage at TRIA applications as variables. Ran- domized complete block designs were used with the exception of the studies on nutrient levels, age of application and method of applica- tion. In these studies, a split-plot design was utilized with nitrogen, age of crop and placement of chemical as the main-plot and TRIA treat- ments as the sub-plots. All field studies consisted of 4-6 blocks. The results of all tests were subjected to analysis of variance and relevant orthogonal and non-orthogonal comparisons were made. 14 Table 6. Cultural conditions of wheat treated with foliage sprays in Central Michigan in 1978. Location East Lansing Saranac Soil type Conover loam Matherton loam Cultivar 'Ionia' 'Yorkstar' 'Ionia' 'Yorkstar' Stage of crop and date Early tiller 4/27 4/27 4/27 4/27 Boot 6/2 6/2 6/3 6/3 Post anthesis 6/27 6/27 6/28 6/28 l/ha 225 225 225 225 Plot size (m) l.2x4.9 l.2x4.9 1.2x4.9 1.2x4.9 Harvest data 8/1 8/1 7/28 7/28 15 Table 7. Cultural conditions of tomato and dry bean crops receiving foliar sprays at various stages of growth in 1978. Crop and Cultivar navy bean 'Sanilac' tomato 'Heinz 1350' Soil type Planting date Planting method Plot size (m) l/ha Fert applied Weed control Harvest date(s) Type of harvest Morphological stage Treatment date(s) Parkhill clay loam 6/10 commercial planter 0.76x15.2 438 336 kg/ha 12-12-12 chloramben and trifluralin 9/20 single lst trifoliate lst pod set 7/7 and 8/8 Spinks sandy loam 5/30 V-belt l.5x7.6 438 589 kg/ha 12-12-12 metribuzin (0.3 kg a.i./ha) 9/29 and 10/10 multiple 1-3 leaves lst flower 6/28 and 7/18 16 Table 8. Cultural conditions of carrots and sweet corn grown on different nitrogen levels. Initial fertility Additional N (ammonium nitrate) 560 kg/ha 12-12-12 67 kg N/ha Crop Carrot Sweet Corn 'Spartan Classic' 'Gold Cup' Year 1977 1978 Planting date 4/14 5/26 Depth of seed (cm) 2 3.8 Planting method V-belt V-belt Treatment date 5/18 6/14 l/ha 984 438 Nozzle size 1-8004 1-8003 .Harvest date 7/6 8/18 Type of harvest single multiple Weed control hoeing atrazine 1.1 kg a.i./ha + alachlor 2.2 kg a.i./ha 560 kg/ha 5-20-20 84 kg N/ha, 168 kg N/ha 17 N H~\m o.ax~.~ awed chmm mxcfimm m mH\w o.nxm.o EmoH Mada: OH w\m o.an.H Emoa chmm camponmo mumo>umz mo nonesz bump umo>um= may oNfim uofim max» Hfiom wm\o.wW\o.om\m mm\n V\m oumv uncapmonh mmouvoVwm .oomMHSm .voom am:\a oomfiv socopp ounmnsm psoEoomHm oflscmnm :OHmH:Eo oflscmum :ofiumaseuom n.H o.v .. nEuV swamp mcflvoom “Hon-> puon-> -- venues wcueoom om\m m~\m va\ofi\v muse menemom whoa nnma mead new» .ommH Nahum. .dsu eHoo. .aoumcngmaz sums. OHNEOH fihco H003m aw>wuasu paw mono mswmhmmmm .wnma vcm 55mg cw mpcmam ecu pcsoum Hwom one ow mcoflumowammm wmoop macho Hmuo>om mo mcofluwvcoo amuauaau .m canmh l8 flan\.n.m we m.ov enaznncpoe o~-o~-m m:\wx com Am;\.u.m mx H.Hv ecunacum NH-~H-~H m:\mx «om moz ezz m:\mx om Houucoo coo: voflnmmm whom .ommH Ncfioz. cameo» .dsu eHoo. 500 90025 sm>fiuasu cam mono .couwcwgmmz xumz. mammemamm .Apozcwucoov .m oflnmh 19 Table 10. Cultural conditions of wheat receiving granular applications of TRIA in 1978. Location East Lansing Saranac Soil type Conover loam Matherton loam Cultivar 'Ionia' 'Yorkstar' 'Ionia' 'Yorkstar' Stage of crop and date Early tiller 4/27 4/27 4/27 4/27 Boot 6/2 6/2 6/2 6/2 Post anthesis 6/22 6/22 6/27 6/27 Plot size (m) 1.2x4.9 1.2x4.9 1.2x4.9 1.2x4.9 Harvest date 8/1 8/1 7/28 7/28 RESULTS AND DISCUSSION Seed Treatments In 1977, seed soak experiments were performed on five different crops. There was no visible response on the subsequent growth of seed- lings from seeds treated with TRIA compared to seeds soaked in DCM only (control). The lack of response to TRIA was also observed in the marketable yields of these crops (Table 11). A different set of experi— ments was carried out at the same time as the previous experiment to determine any difference between the effect of TRIA applied as a seed soak to alfalfa placed around the seeds during planting. TRIA again did not increase the yield of the crops, however, there was an increase in yield from seed soaking in DCM alone with the tomatoes (Table 12). Contrary to expectation, planting the seed with alfalfa did not in- crease the yield of sweet corn and tomato, while barley yields were de- creased. In 1978, DCM or TRIA and DCM did not produce plants having higher yields than the controls when directly seeded in the field (data not presented). It can be concluded that soaking seed of crops in a TRIA-DCM solu- tion for direct seeding in the field is not a feasible means of TRIA application. Applying TRIA to the seed used for the production of field transplants proved to be the only effective method of application to the seed in 1978. The fresh weight of transplants from TRIA-treated seedlings 20 21 Table 11. Yield of several vegetable crops when seed soaked in DCM- TRIA solution for one hour in 1977. Yield (metric tons/ha) TRIA (mg/l) Cucumber Carrot Lettuce Radish Sweet corn Control 20. 24.6 14. 2.0 13.2 DCM Control 19. 24.1 11. 2.0 13.2 .01 20. 24.2 16. 1.9 11.5 .1 20. 24.1 15. 1.8 12.2 1.0 19. 23.2 15. 1.9 12.6 10.0 20. 23.7 12. 1.9 12.5 Coefficient of variation (%) 9. 5.1 18. 7.5 13.5 22 .Ho>oH Ho.o um ucmonHcmHm Zoo 0: .m> comm uopmohp Zoo pom osHm> m\N .Ho>oH mo.o um pconchmHm mmHmmHm csz pHon :H ommogoou HmocHH how osHm> m.l \N o.nm v.om Hn.m m.wH oH.o oH.o oH.o Hom o.vm m.m mm.~ m.wH oH oH oH mmHmmH< m.mm m.v No.m n.wH H H H mmHmmH< m.mm N.c 0H.m m.mH o o o Hahucou oumH xHumo \monumn :Hoo poozm HH\wEV mm:\mxv HH\MEV mm:\wxv HH\mEV Hm:\mxv oosmumnsm \NOHmEou oumaou onHmn choc poozm Hm;\m:ou oHupmeV eHoH» QHNH HGfiEHMOHB .nan :H mcmeom om mo vHoH> .NH oHHMH 23 (8.0 g/plant) was larger than the non-soaked (5.1 g/plant) and DCM soaked (4.8 g/plant) seed at time of transplanting to the field. This TRIA response was maintained at harvest with the plants from TRIA-DCM treatments yielding 27% more fruit than the controls (Figure I). TRIA at 50 and 500 mg/ha as a foliage application and 400, 1600, 6400 mg/ha as a granular application, were made to the plants when flowers first appeared in the field. These post—applications increased the yield 13% over non-treated controls, however, a yield decrease of 12% occurred when plants from pretreated seed received post-applications. This is the only study in either year that shows a decrease in yield from multiple applications of TRIA. Placement of the granular formulation of TRIA with the seed of cucumbers also was not an effective mode of application (Table 13). The yield of non-treated plots was significantly larger than the granular blank plots. Therefore, the granules, when placed with the seed, had a detrimental effect on cucumber yield. However, a comparison of all TRIA treatments to granular blank showed significantly higher yields for TRIA plots indicating that this effect was overcome by TRIA. Foliar Treatments Applying TRIA as a foliage spray when the plants had 2-3 true leaves proved to be an effective method of treatment for several crops in both years of study. However, in only a few cases could increased growth be observed; for example, the average height of all sweet corn treatments was 13% higher than the controls 15 days after treatment (Table 14). The overall yield of sweet corn was increased 16% by TRIA applications, this increase could not be attributed to either ear number 24 Figure 1. A comparison of 'Heinz 1350' tomato yield from TRIA treatments applied to the seed and to plants after flowering in the field. YIE LD (tons/ acre) 30 I0 0 1O —— ~ ~ ~ * — ~ ~ —— ~ * —— u — — —— — — — — — _ — — _ —_ — —_ —_ —-I- — — —— — — — — — — — fl — — — — — _ — — — — — _ — — — _ _ — — — — —_ — — _ — — * ~ — * — I.— — * —_ — — — — — — — _ 3 O 3 0 N U1 * I—_ — — — —_ —— —I— _ — — — — — —— — —— —— —_ _— —_ __ —_ —_ __ — — — —I— — —_ — — — — — — — — — — _ — — ~ ~ —— — * ——- — ~ ~ ~ I—h * -_ _ ~ —I_ _ * fi * — * * _— —_ — ~ — * — — — _ _ — — _ — _ _ — —_ _ — — — — — — — —— — flowering seed & flowering TIME OF APPLICATION seed 26 Table 13. Response of 'Green Star' pickling cucumbers to TRIA applied as a granular formulation with the seed in 1978. TRIA Yield (mg/ha) (mt/ha) -- 15.03! Granular blank 11.6 40 14.1 160 15.6 640 13.8 2560 13.0 Coefficient of variation (%) 11.4 L.S.D. at 0.01 level 2.5 2/ -F value for granular blank vs. TRIA treatments is significant at the 0.01 level. 27 .Ho>oH mo.o on» um pcmoHMchHm :omHHmanu oHumnpmsc how 03Hm> m.l \> .muon mm pom: one: mucmHn HmsvH>Hp=H\m .Ho>oH Hc.o ecu um ucmonchHm :OmHHmmaoo oHumhvac Mom ous> m.l \x .HQ>0~ OHQN How pom: mHouuaoo mo owmuo>m cos: .Ho>oH mo.o any we ucmonchHm m\N .Ho>oH Ho.o on» an acmonHanm Honucoo ou wohmmaou mucoEumouu pom osHm> m\m -- -- -- eo.~ -- -- -- oooH -- -- -- co.» -- -- -- ooH moH mm.oH -- om.~ -- mm.NH mm o.oH moH ON.HH mm.H -- am.mH mo.NH mm o.H oHH me.HH ew.H -- Nm.m~ em.mH oe H.o -- -- -- -- mo.om -- -- mo.o - EHH mw.HH HE.H -- we.eH Hm.HH mm Ho.o -- -- Hw.H -- mH.HN -- -- Houucou conH=Em \Mmm \Mwm.a \NHw.H \HEE.H mm.eH \m©a.oH \mNm Honueou HuoHd\o=L Ha;\usv Hae\uev Hpcde\mxv Hm:\usv Hme\psv Hauv HH\wev uHsum a: pamHa HoooH macho Hmuo>om mo pHoH> .vH oHnmh 28 or weight/ear alone, but a combination of these factors. Cucumber yield was increased an average of 13% by TRIA applications and could be attributed to an increase in number of fruit (13%) in treated plots. Yields of radishes exhibited a linear trend with increasing rates of application. Since the yield of fruit from individual tomato plants sprayed with high or low rates of TRIA was similar, it can be concluded that high rates of TRIA (1000 mg/l) did not injure plants. The effect of multiple applications of TRIA was studied on cucumber, navy bean and tomato plants. The crops were treated initially at the young seedling stage, with the exception of the tomatoes which received initial treatments at first flowering, and received additional applica- tions at weekly or bi-weekly intervals throughout the growing season. Rates of 0.01, 0.10 and 1.00 mg/l were applied to the navy beans and cucumbers while the tomatoes received rates of 1.0 and 10.0 mg/l through- out the season. These rates were applied with each of the application treatments on different plots, but there was no significant difference between rates and there was no interaction of rates with number of applications (Table 15). The navy bean and cucumber plots treated with TRIA compared to controls exhibited 7% and 11% yield increases for each crop. Sprays applied to transplanted tomatoes at different nitrogen levels increased the early yield by 14% while having no effect on late yield. There was no significant difference in yield at the two nitro- gen levels and no interaction of TRIA application with nitrogen levels. These studies of crops which received multiple applications of TRIA showed no greater increase of growth than from a single application. There were no clear differences in yield at the rates applied in these studies. 29 Table 15. The yield of several crops receiving single or multiple applications of TRIA in 1977. tomato 2 / x/ 'Heinz 1350' cucumber—LX- dry bean—- (mt/ha) Number of 'Green Star' 'Sanilar' x/ applications (mt/ha) (mt/ha) early—- late Control 14.72 2.41 9.40 30.0 Single 16.01 2.64 11.03 32.9 Multiple 16.58 2.56 10.53 30.9 E-/F value for control vs. treatment significant at the 0.01 level. X/F value for linear trend with increasing number of applications signi- ficant at the 0.01 level. x/ -— F value for control vs. treatment significant at the 0.05 level. 30 Wheat, navy bean and tomato crops were treated with TRIA at dif- ferent stages of growth in an attempt to determine the optimum morpho- logical stage of growth for application. In the wheat tests, foliar and granular treatments were applied at early tiller, boot and post- anthesis stages of growth. There were no significant differences due to formulation or time of application (Table 16). There was a quadratic trend for the increase in yield with different rates fer the different cultivars at both locations. At East Lansing, there was an increase in yield from TRIA applications for 'Yorkstar', but not 'Ionia'. At the Saranac location, there was an increase in yield from TRIA applica- tions for 'Ionia', but not 'Yorkstar'. The yields of navy bean and tomato crops treated in early vegetative and reproductive stages of growth was not significantly increased, yet the data would indicate a response to TRIA (Table 17). There was no interaction or rate with time of application in either crop. The same rates applied to cucumbers at the 2nd-leaf stage of development, increased the yield as indicated by a cubic trend with increasing rates of application. The data from the three crops (Table 17) were analyzed as percent of controls and split for crops. Controls were averaged in the three crops. The data for each rate of application and age were averaged in the tomato and navy bean crops. The average of all treatments, 105%, versus controls was significantly higher and the tomato crop responded significantly more to TRIA than either the cucumber or navy bean crops. Nitrogen studies were conducted with sweet corn and carrots. Yields of carrots indicate that when TRIA was applied at low nitrogen levels there was a linear increase in yield ranging from 11% to 21% when compared to controls (Figure 2). However, with applications to carrots 31 Table 16. Response of wheat cultivars to TRIA applied in different formulations, times and rates at Saranac and East Lansing in 1978. Each observation is the average for the different times and rates of applications. Yieldz/ Treatment z/ East Lansing Saranac Cultivar TRIA—- (bu/a) (bu/a) 'Ionia' 0 50.5 46.4 low 48.0 50.1 medium 48.5 49.3 high 49.7 48.2 'Yorkstar' 0 49.8 35.0 low 50.8 35.4 medium 51.3 34.2 high 47.6 35.3 E-/Low, medium and high rates were 10, 40 and 160 and 40, 400 and 1600 mg TRIA/ha, respectively, for the emulsion and granular formulations. y/ —-F value for the quadratic trend with different rates for the cultivars was significant at the 0.05 level at both locations. 32 Table 17. Yield of cucumber, tomato and navy bean crops when treated with foliar sprays of TRIA in 1978, and average yield analyzed ag/percent of controls in an analysis with crop as main plot;- rx/ cucumbe navy bean tomato % of control TRIA 'Green Star' ‘Sanilac' 'Heinz 1350' x/ (mg/ha) (mt/ha) (mt/ha) (mt/ha) for 3 crops—- Controls 17.1 2.00 32.33 100 2.5 19.3 1.98 34.21 106 10.0 17.6 2.05 35.45 105 40.0 16.6 2.06 34.52 103 160.0 17.4 2.12 35.81 107 2/ -The yield from tomato and navy bean plots treated at early vegetative reproductive stages of growth was combined because there was no significant interaction of rate with age of application. Z/F value for cubic trend significant at the 0.05 level. X/ F value for treatments vs. controls significant at the 0.01 level. 33 Figure 2. Yield of 'Spartan Classic' carrots treated with TRIA at two different N levels. P value for linear trend in yield with different nitrogen levels significant at 0.05. 34 (D b 33 ,. 32 ' 67ng/ha 31 3o N (D O YIELD (MT/ha) N N \I oo v=2a;-.°,:;,-«..J,2,?,'fg”Ta l 0 u. 26 ‘ 25 0.0 0.01 0.10 1.00 TR IA (mg/ I) 35 growing under the higher N level there was a linear decrease in yield. In the sweet corn test with three N levels there was a quadratic rate response of TRIA with a linear rate of N supplied to the crop. The highest yields occurred at the low N level with the higher rates of TRIA and at the high N level with low rates of TRIA application (Figure 3). Soil Treatments Treatments were applied around the base of sweet corn plants with a high volume of TRIA solution to stimulate a soil drench. There was no response to the soil treatments (data not presented). The effect of TRIA on asparagus was also studied. The high rate of TRIA granules (1.6 g/ha) applied to the soil surface increased the early yield 33% (Table 18). This rate when compared to all other treatments at different harvest times was significant at the % level. Neither the weight/spear nor the number of spears were solely responsible for the yield increase. In two other studies conducted in Oceana County, involving TRIA applications to asparagus, increases in yield. were not observed. Granular formulations of TRIA were applied with the seed, side- dressed (6.4 cm x 6.4 cm from seedlings) and on the soil surface of tomato plants. There was no significant increase in yield and no inter- action of treatment with method of application (Table 19). However, inadequate controls were performed in this experiment since granular blank applied with the seed may be detrimental to yield (Table 13). 36 Figure 3. Yield of 'Gold Cup' sweet corn at different rates with different levels of N nutrition. F value for quadratic rate of TRIA with linear rate of nitrogen significant at 0.01 level. 37 .000. a .‘l! a 00 .n - a h .0 / .. h M o... N I. W o. 9 7 9 00 .K - 9 k 000 4 .. k 8 0000 8 .. .. 8 2 00000 . O. m 0000‘ . .. .0.- O. 7. . .00 - . 000 .7 .. 000 O. . 000 - . 00 .. H 0000 O. 00 C... .41 . ... 1”, O... C ’1’, .0. It .. III ... It ... . 0 "st,” 0.. .. .4 I , .. I r l I . l 0 9 n m w. u m n n 7 6. 3.0m \mcot 04m. > high low RATES OF TRIA 38 Table 18. A comparison of yield of "snapped" 'Mary Washington' asparagus after treatment with TRIA at Sodus, Michigan in 1978. . z/ . Yield (g/plot)—- Weight (g/spear) TRIA Total Total Formulation (g/ha) Early Late Early Late Control 0 616 308 14.0 15.7 Emulsion 0.04 661 236 14.4 13.9 Granular 0.40 671 313 16.2 12.7 Granular 1.60 819 228 14.8 14.5 2/ - F value for difference in yield between high rate of granular and other treatments at different harvest times significant at the 0.05 level for yield. 39 Table 19. Yield of 'Heinz 1350' direct seeded tomatoes as influenced by granular applications of TRIA. Method of TRIA Yield application (mg/ha) (mt/ha) Seed Granular blank 37.7 400 . 45.9 1600 39.3 Sidedressed Granular blank 37.0 400 38.6 1600 39.2 Surface Granular blank 42.9 400 39.1 1600 40.1 Coefficient of variation (%) ' 14.3 CONCLUSION Visual effects of TRIA were often not observed in the field, al— though increases in yield were measured. This may be due to biological variation among plants grown in the field since differences in growth were observed under the more uniform conditions of the greenhouse. Foliar sprays appear to be the most effective method of TRIA application. Foliar applications of TRIA from 20 mg/ha of crop to 2.2 g/ha were especially effective in increasing yield in the field applications. While there was no clear rate range, the plants response to TRIA is generally more consistent at lower concentrations of TRIA. High concen- trations of TRIA (1000 mg/l) were not harmful to tomatoes. Single appli- cations of TRIA were as effective as multiple applications. The morpho- logical stage of growth did not affect the plants response to TRIA. Seed soak applications of TRIA, while very effective in the green- house, do not increase growth of plants in the field, unless the seed- lings are grown for field transplants. The granular formulations of TRIA may increase the growth of the crops and more research is necessary to determine the best placement of the granules near the seed. Surface applications increased the yield of several crops, but it would appear that higher rates of TRIA are re- quired for the yield response. Nitrogen studies indicated that crops respond to a greater degree to TRIA when grown under low nitrogen regimes. 40 41 From observations of the 0.1% emulsion, it is thought that a greater effort in formulation is required as the concentrate is not stable at room temperature. Whether the greater variation was due to environment or cultural practices is not certain. Unpublished data (Ries, Wert) from greenhouse studies indicate that volume delivered may be a critical factor in the plants response to TRIA. This informa- tion may relate to the success of field applications in 1978, since sub- stantially lower volumes were applied the second year of study. TRIA, under conditions which were not made clear in this study, will increase the yield of several crops. This increase is not signi- ficantly consistent to suggest that TRIA be used by growers. More research must be conducted to determine the optimum parameters for commercial use of TRIA in the field. 10. 11. 12. LITERATURE CITED Bittenbender, H. C., D. R. Dilley, V. F. Wert and S. K. Ries. 1978. Environmental parameters affecting dark response of rice seedlings to triacontanol. Plant Physiol. 61:831-854. Chibnall, A. C., E. F. Williams, A. L. Latner and S. H. Piper. 1933. The isolation of n-triacontanol from lucerne wax. Biochem. J. 27:1885-1888. Hangarter, R., S. K. Ries and P. Carlson. 1978. Effect of tria- contanol in plant cell cultures in_vitro. Plant Physiol. 61:855-858. Jones, J. F., V. F. Wert and S. K. Ries. 1979. Specificity of l- triacontanol as a plant growth stimulator and inhibition of its effect by other long-chain compounds. Planta 144:277-282. Kolattukudy, P. E. and T. J. Walton. 1972. The biochemistry of plant cuticular lipids. Prog. Chem. 13:121-175. Kolker, L. 1978. Analytical procedures for l-triacontanol and its presence in plants and the environment. M.S. Thesis, Michigan State University. Martin, J. T. and B. E. Juniper. 1970. The cuticles of plants. Edward Arnold Ltd., N.Y., pp. 347. Ries, S. K., H. Bittenbender, R. Hangarter, L. Kolker, G. Morris and V. Wert. 1976. Improved growth and yield from organic supple- ments. In; W. Lokeretz, ed. Energy and Agriculture. Acad. Press, N.Y., Pp. 377-394 (1977). Ries, S. K., T. L. Richman and V. Wert. 1978. Growth and yield of crops treated with triacontanol. Amer. Soc. Hort. Sci. 103: 361-364. Ries, S. K. and V. Wert. 1977. Growth responses of rice seedlings to triacontanol in light and dark. Planta 135:77-82. Ries, S. K., V. Wert, C. C. Sweeley and R. A. Leavitt. 1977. Triacontanol: A new naturally occurring plant growth regulator. Science 195:1339-1341. * Waldron, J. D., D. S. Gowers, A. C. Chibnall and S. H. Piper. 1961. Further observations on the paraffins and primary alcohols of plant waxes. Biochem. J. 78:435-442. 42 ”‘IIIIIIIIIIIIIIEs