Luận văn effect of low temperature on the germination of cucumber seeds

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY DO THI QUYNH TRANG EFFECT OF LOW TEMPERATURE ON THE GERMINATION OF CUCUMBER SEEDS BACHELOR THESIS Study mode: Full-time Major: Environmental Science and Management Faculty: Advanced Education Program Office Batch: 2014 - 2018 Thai Nguyen, 16/09/2018 h DOCUMENT PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Degree program Bachelor of Environmental Science and Management Student name Do Thi Quynh Trang Student ID DTN1454290031 Effect of low temperature on the germination of cucumber Thesis Title seeds Dr Duong Van Thao Supervisor (s) Assist Prof San-Gwang Hwang Supervisor’s signature (s) Abstract: Poor seed germination is a common phenomenon at sub-optimal temperatures which causes a great concern for farmers Several priming treatments have been reported to test for the seed resistance at different temperatures, particularly at low temperatures In this study, six cucumber cultivars (CU-87, CU127, Cuigu, Kappa summer no 7, Kappa summer no 11and CU-74) were used to perform in four different temperatures (10℃, 15℃, 20℃ and 25℃), to compare the germination and cold tolerance of seeds under low temperature conditions There are five experiments were performed and each experiment was run in three replications The results showed that cucumber was not suitable at low temperature and tolerance of cucumber in poor chilling conditions The germination was highest at 25℃ and 20℃ but seeds also germinated readily at 15℃ and no germination was observed at 10℃ Pre-treatment of seeds in hot water for 10 minutes had only minor ii h effect on germination rate Key words: Low temperature, chilling, germination, low treatment, rate root, cucumber Number Pages: 45 Date of Submission: 16/09/2018 iii h ACKNOWDLEDGEMENT I would like firstly to emphasize the sincere appreciation to teachers in Advance Education Program Office as well as teachers in Thai Nguyen University of Agricultural and Forestry, who have taught me knowledge not only for my subjects but also for my living skills and gave me a chance to my thesis abroad In addition, I would like to thank all supports and help from Department of Horticulture, National Chung Hsing University for the time I did my research in Taiwan It is my pleasure to work with a great teacher - Assistant Professor San-Gwang Hwang, who always helped me any time He also gave me the best conditions, supported all materials for my research and discussed about any problems I got whenever I did experiments in his Vegetable Laboratory Indeed, without his help this study would not have been possible I am thankful that he provided me an opportunity to establish my knowledge I would like to give special thanks to Dr Duong Van Thao, who always supported and cheered me up whole the time I worked over-sea He also helps me a lot on spending much time for checking my thesis report I sincerely respect him for his devotion to work, simplicity, his care, cooperation and encouragement during the study period I consider it is an honor to work with Mr Wayne, a master student, who particularly helpful in guiding me toward a qualitative methodology and inspiring me in whole period of internship time He was always helpful, friendly and very kind with me Without his guidance, I cannot complete this thesis iv h Finally, I would like to express my gratitude to my family and friends, who always beside me all the time Their helps, supports and encouragement created the pump leading me to success Sincerely, Do Thi Quynh Trang v h TABLE OF CONTENT ACKNOWDLEDGEMENT iv TABLE OF CONTENT vi LIST OF FIGURES viii LIST OF TABLES viii LIST OF ABBREVIATIONS ix PART I INTRODUCTION 1.1 Research rationale 1.2 Research objectives 1.3 Research questions and hypothesis 1.3.1 Research questions 1.3.2 Hypothesis .3 1.4 Limitations 1.5 Definition PART II LITERATURE REVIEW 2.1 An overview about the experiment 2.2 Factors influence the germination of seeds 2.2.1 Temperature 2.2.2 Ethylene (C2H4) and respiration production (CO2) 10 2.2.3 Cell membrane on the germination of seeds .12 PART III METHODS 15 3.1 Materials 15 3.2 Methods 16 vi h 3.2.1 Germination test 16 3.2.2 Root growth (Radicle length) 17 3.2.3 Relative electrical conductivity (EC) 17 3.2.4 Respiration measurement (CO2) and ethylene measurement (C2H4) 17 3.2.5 Statistics .18 PART IV RESULT .19 4.1 Final germination percentage (FGP), days to 50% germination and 80% germination .19 4.2 Root growth (Radicle length) 21 4.3 Relative electrical conductivity (EC) .27 4.4 Respiratory production rate (CO2) 29 4.5 Ethylene production rate (C2H4) 32 PART V DISCUSSION AND CONCLUSION 35 5.1 Discussion………………………………………………………………………………………….….35 5.2 Conclusion……………………………………………………………………………………….……36 REFERENCES 38 vii h LIST OF FIGURES Figure 4.1 Effect of temperature on root growth for days 24 Figure 4.2 The root of cultivars at 10℃ after days 25 Figure 4.3 The root of cultivars at 15℃ after days 25 Figure 4.4 The root of cultivars at 20℃ after days 26 Figure 4.5 The root of cultivars at 25℃ after days 26 Figure 4.6 Effects of tolerance on relativity electrolyte conductivity 29 Figure 4.7 Seeds respiration rates at different temperature 31 Figure 4.8 Ethylene production rate change in seed within low temperature 34 viii h LIST OF TABLES Table 3.1 Cucumber seeds information 16 Table 4.1 Germination ability at four temperatures in six cucumber cultivars 19 Table 4.2 Effect of germination temperature on the time to 50% germination 21 Table 4.3 Effect of germination temperature on the time to 80% germination 21 ix h LIST OF ABBREVIATIONS CO2 The respiration C2H4 The ethylene EC Electrical conductivity EC0 Electrical conductivity of the seeds after put in different temperature levels for 24 hours EC1 Electrical conductivity of the seeds after bath in 95℃ hot water for hour x h 4.5 Ethylene production rate (C2H4) Ethylene can rouse germination and overcome dormancy in many seeds The participation of ethylene in seed germination is a widely accepted fact, but the mechanistic details are poorly understood Ethylene production by seeds starts immediately after the onset of imbibition and increases with time However, the pattern of ethylene production by seeds during germination differs among species 32 h 33 h Figure 4.8 Ethylene production rate change in seed within low temperature The results indicate that in this experiment no C2H4 was involved during the germination of cucumber seeds at 10℃ and 15℃ for 48 hours (Figure 4.8) The osmopriming of the seed influence C2H4 production during seed germination, at a low level of heat that falls into dormancy and can not germinate Seed dormancy prevents germination during periods unfavourable to seedling growth (Matilla, 2007) Conversely, we can see that high temperatures of 20℃ and 25℃ have the appearance of C2H4, in particular, the 20℃ ethylene germination parameters in the germination process is higher than the ethylene in 25℃ in out of cultivars (Figure 4.8) This pointed out that at high temperatures of 25℃ also lead to reduced C2H4 production In research on carrot seeds (Nascimento et al., 2013) indicated that for primed seeds, C2H4 production was lower at 35℃ than at 20℃ High temperatures have been shown to inhibit C2H4 production in seeds Ethylene production would be highly effective as long as they were germinated under optimum conditions, and in this experiment we showed that the suitable temperature for C2H4 production in cucumber seeds was 20℃ 34 h PART V DISCUSSION AND CONCLUSION 5.1 Discussion Cucumber seeds show similar responses to chilling as determined by the parameters tested in this study This plant were sensitive during germination time at low temperature rather than in proper temperature Seeds used for conduct experiment at under 15℃ are considered to be sensitive with cold temperatures, which made the seeds not only can not germinate but also cause the root systems were not fully develop The roots of the pre-treated seeds, placed in refrigerators with levels of temperature is 10℃ and 15℃ can germinate, however, the root length is not more than cm at 10℃ and results in 15℃ is not longer than cm (Figure 4.2 and 4.3) This result because of cold temperature that prevents the cucumber seeds from absorbing enough water and nutrients even though they was placed under laboratory conditions In many previous studies have shown that the lower the temperature the higher will damage the cell membrane and lead to the high rate of EC Studies on soybean seeds have shown that the results of the EC test may be influenced by storage temperature, especially low temperatures such as 10ºC (Vieira et al., 2001) However, in this report, the results showed that the percentage of EC at all four heat levels was the same, over a period of day of cooling; the membrane did not have too much difference due to the cold different heat levels Working with soybean seeds, (Vieira et al., 2001) mentioned that membranes also stabilized for seeds stored at 10℃, resulting in no increase in conductivity This can raise some questions Why should seeds stored at 10℃ apparently stabilize membranes more than seeds stored at 30℃, for instance, 35 h resulting in lower value of EC? According to (Bernal-Lugo & Leopold, 1998), the transition from a period of relative membrane stability to dynamic seed aging could occur through a loss of the glassy state This loss could be influenced by an increase in the water content, in temperature, or by a separation of sugars involved (Hu et al., 2006) Besides, the beginning of deterioration could result from a gradual hydrolysis of the soluble sugars In cellular CO2, the seeds use stored sugars, water and oxygen to burn energy at a cellular level and germinate, or sprout Respiration increases dramatically as the seed sprouts So at low temperatures the affected seeds can only breathe at a very low frequency, meaning that cucumber seeds can not absorb enough energy to burn off and germinate Conversely, high temperatures will allow the CO2 process to take place fully, stimulating the germination of the seeds These results are consistent with the effects of suboptimal temperature on most biological processes in plants (Dahal et al., 1996) Being the majority of seed germination, C2H4 production does not occur at low temperature because cold temperatures cause seeds to be suppressed only after 48 hours of cooling It demonstrates that cold temperatures strongly affect the ethylene production of the seed and become one of the contributing factors to seed deterioration under severe conditions 5.2 Conclusion Seeds play an important role in nature, agriculture and horticulture Seeds are a special plant structure with specific properties and functions Simple in their structure and design, seeds are very species specific Seed germination is a complicated process 36 h and it is regulated by many factors such as nutrient, temperature, water, light and substrate However, temperature plays an integral role in the whole of this process This study tried hard to analyze the results of the studies on the problem of cucumber seed germination under different temperature treatments, and found that, as the lower the temperature, the lower the cucumber seed's germination ability, these two factors are proportional Temperature does not only affect the development of the root system and the membrane, but it also has a major impact on the respiration of the seed during germination Proper temperature will help the seed to germinate well to produce high yield, whereas cold temperatures affect the growth and development of the seed, the longer cold duration, the greater the risk to the seed The result of this study shows that seeds grew fastest and strongest from 20℃ to 25℃, however it was harmful for the seeds at 10℃ and 15℃ 37 h REFERENCES A van de Venter, H., and Grobbelaar, N 1985 Influence of sub-optimal imbibition temperatures on seed vigour and respiration in maize ( Zea mays L.) South African Journal of Plant and Soil, (pp 203-206) ALVAREZ‐URIA, P and KÖRNER, C 2007, Low temperature limits of root growth in deciduous and evergreen temperate tree species Functional Ecology, (pp 211-218) Balkaya A., Kurtar ES and Cemek B 2008 Modelling the effect of temperature on germination power in some Brassica species African Journal of Biotechnology Vol (9), (pp 1343-1353) Bernal-Lugo, I., and Leopold, A C 1998 The dynamics of seed mortality Journal of Experimental Botany, 49(326), (pp 1455-1461) Bewley, J D 1997 Seed Germination and Dormancy The Plant Cell, 9(7), (pp 10551066) Bewley, J D., Bradford, K J., Hilhorst, H W M., and Nonogaki, H 2013 Environmental Regulation of Dormancy and Germination Seeds Physiology of Development, Germination and Dormancy, 3rd Edition (pp 299-339) New York, NY: Springer New York Crowe, J H., and Crowe, L M 1992 Membrane Integrity in Anhydrobiotic Organisms: Toward a Mechanism for Stabilizing Dry Cells Water and life, (pp 87-103) Springer, Berlin, Heidelberg 38 h Cheng, L.-b., Li, S.-y., and He, G 2009 Isolation and Expression Profile Analysis of Genes Relevant to Chilling Stress During Seed Imbibition in Soybean [Glycine max (L.) Meer.] Agriculture science, 8, (pp 521-528) Christiansen, M N 1967 Periods of Sensitivity to Chilling in Germinating Cotton Plant Physiology, 42(3), (pp 431-433) Cırak C, Ayan AK, Odabas MS and Camas N 2007 Modelling the effect of temperature on the days to germination in seeds of flue-cured and oriental tobacco (Nicotiana tabacum L.) Journal of Plant Sciences, 2, (pp 358-361) Dahal, P., Kim, N.-S., and Bradford, K J 1996 Respiration and germination rates of tomato seeds at suboptimal temperatures and reduced water potentials Journal of Experimental Botany, 47(7), (pp 941-947) Devaiah, S P., Roth, M R., Baughman, E., Li, M., Tamura, P., Jeannotte, R., Wang, X 2006 Quantitative profiling of polar glycerolipid species from organs of wild-type Arabidopsis and a Phospholipase Dα1 knockout mutant Phytochemistry, 67(17), (pp 1907-1924) Dürr, C., Aubertot, J N., Richard, G., Dubrulle, P., Duval, Y., and Boiffin, J 2001 Simple: A model for SIMulation of PLant emergence predicting the effects of soil tillage and sowing operations Soil Science Society America Journal Abstract, 65, (pp 414-442) Fessel, S A., Vieira, R D., Cruz, M C P d., Paula, R C d., and Panobianco, M 2006 Electrical conductivity testing of corn seeds as influenced by temperature and period of storage Pesquisa Agropecuária Brasileira, 41, (pp 1551-1559) 39 h Finér, L., Helmisaari, H.-S., Lõhmus, K., Majdi, H., Brunner, I., Børja, I., Möttönen, M R 2007 Variation in fine root biomass of three European tree species: Beech (Fagus sylvatica L.), Norway spruce (Picea abies L Karst.), and Scots Pine (Pinus sylvestris L.) Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology, 3, (pp 394-405) Flores, J., and Briones, O 2001 Plant life-form and germination in a Mexican intertropical desert: effects of soil water potential and temperature Journal of Arid Environments, 47(4), (pp 485-497) Francisco, G., Katharina, D., Isabel, D., Ulrich, Z., Nicolas, G., Peter, D., and Dorothea, B 2013 The role of lipid metabolism in the acquisition of desiccation tolerance in Craterostigma plantagineum: a comparative approach The Plant Journal, 75(5), (pp 726-741) Håkansson, I., Myrbeck, Å., and Etana, A 2002 A review of research on seedbed preparation for small grains in Sweden Soil and Tillage Research, 64(1), (pp 23-40) Hardegree, S P 2006 Predicting Germination Response to Temperature I Cardinaltemperature Models and Subpopulation-specific Regression Annals of Botany, 97(6), (pp 1115-1125) Hardegree, S P., Flerchinger, G N., and Van Vactor, S S 2003 Hydrothermal germination response and the development of probabilistic germination profiles Ecological Modelling, 167(3), (pp 305-322) 40 h Hobbs, P R., and Obendorf, R L 1972 Interaction of Initial Seed Moisture and Imbibitional Temperature on Germination and Productivity of Soybean1 Crop Science, 12, (pp 664-667) Hu, W H., Shi, K., Song, X S., Xia, X J., Zhou, Y H., and Yu, J Q 2006 Different effects of chilling on respiration in leaves and roots of cucumber (Cucumis sativus) Plant Physiology and Biochemistry, 44(11), (pp 837-843) Index 1992 In F B Abeles, P W Morgan, and M E Saltveit (Eds.), Ethylene in Plant Biology (Second Edition) (pp 399-414) New York: Academic Press Irwin, C.C and H.C Price 1983 The relationship of radicle length tochilling sensitivity of pregerminated pepper seed Journal of American Society of Horticultural Science, 108, (pp 484–486) Ishibashi, Y., Koda, Y., Zheng, S.-H., Yuasa, T., and Iwaya-Inoue, M 2013 Regulation of soybean seed germination through ethylene production in response to reactive oxygen species Annals of Botany, 111(1), (pp 95-102) Jenning, P and Mikal E Saltveit 1994 Temperature Effects on Imbibition and Germination of Cucumber (Cucumis sativus) Seeds Journal of the American Society for Horticultural Science American Society for Horticultural Science 119(3), (pp 464-467) Mann Laboratory, Department of Vegetable Crops, University of California, Davis CA 956168631 Jame, Y W., and Cutforth, H W 2004 Simulating the effects of temperature and seeding depth on germination and emergence of spring wheat Agricultural and Forest Meteorology, 124(3), (pp 207-218) 41 h Kłosińska, U., Kozik, E U., Nowicki, M., and Wehner, T C 2013 Low temperature seed germination of cucumber: genetic basis of the tolerance trait Journal of Horticultural Research 21(2), (pp 125-130) Körner, C 2008 Winter crop growth at low temperature may hold the answer for Alpine treeline formation (Vol 1) Körner, C 2011 Coldest places on earth with angiosperm plant life Alpine Botany, 121(1), (pp 11-22) Kozareva, I., Cantliffe, D J., Nagata, R T., and Klee, H J 2004 New support fot the involvement of Ethylene in lecttuce germination at supra-optimal temperature Acta Horticulturae 631, (pp 31-37) Kurtar ES, Balkaya A and Uzun S 2004 Modelling the temperature on the germination percentage in some legume crops Journal of Agronomy, 3, (pp 179-183) Kurtar, E 2010 Modelling the effect of temperature on seed germination in some cucurbits African Journal of Biotechnology 9(9), (pp 1343-1353) Lower R.L 1975 Measurement and selection for cold tolerance in cucumber Pickle Pack Science, 4, (pp 8-11) Lyons, J M 1973 Chilling Injury in Plants Annual Review of Plant Physiology, 24(1), (pp 445-466) Lyons, J M., and Raison, J K 1970 Oxidative Activity of Mitochondria Isolated from Plant Tissues Sensitive and Resistant to Chilling Injury Plant Physiology, 45(4), (pp 386-389) 42 h Ismail, M., Mullen, R., R Stewart, C., and Knapp, A 1989 Respiratory Rates and Alternative Pathway Capacity during Early Germination of Soybean (Vol 29) Makeen, M., Noor, N., Dussert, S., and Clyde, M M 2006 Seed moisture characteristics in relation to total lipid content of five Citrus taxa using an equilibrium dehydration protocol (Vol 34) Malcolm, P J., Holford, P., McGlasson, W B., and Newman, S 2003 Temperature and seed weight affect the germination of peach rootstock seeds and the growth of rootstock seedlings Scientia Horticulturae, 98(3), (pp 247-256) Mary E Mangrich and Mikal E Saltveit 2000 Effect of chilling, heat shock, and vigor on the growth of cucumber (Cucumis sativus) radicles Physiologia Plantarum, 109(2), (pp 137-142) Matilla, A J 2007 Ethylene in seed formation and germination Seed Science Research, 10(2), (pp 111-126) Mayer, A M., and Poljakoff-Mayber, A 1965 The Germination of Seeds (Vol 2) Mazor, L E A., Perl, M., and Negbi, M 1984 Changes in Some ATP-Dependent Activities in Seeds during Treatment with Polyethyleneglycol and during the Redrying Process Journal of Experimental Botany, 35(8), (pp 1119-1127) Mucha, J., Szymańska, A K., Zadworny, M., Tylkowski, T., Michalak, M., and Suszka, J 2015 Effect of seed storage temperature on fine root development and mycorrhizal colonization of young Populus nigra seedlings Annals of Forest Science, 72(5), (pp 539-547) 43 h Nascimento, W M., Huber, D J., and Cantliffe, D J 2013 Carrot seed germination and ethylene production at high temperature in response to seed osmopriming Horticultura Brasileira, 31, (pp 554-558) Nerson, H 2007 Seed production and germinability of cucurbit crops Seed Science and Biotechnology (Vol 1) Odabas MS and Mut Z 2007 Modelling the effect of temperature on percentage and duration of seed germination in grain legumes and cereals American of Journal Plant Physiology, 2, (pp 303-310) Patanè, C., Valeria Cavallaro, Giovanni Avola and Giuseppina D’Agosta 2006 Seed respiration of sorghum [Sorghum bicolor (L.) Moench] during germination as affected by temperature and osmoconditioning Journal of Seed Science Research, 16(4), (pp 251-260) Pollock, B M 1969 Imbibition Temperature Sensitivity of Lima Bean Seeds Controlled by Initial Seed Moisture Plant Physiology, 44(6), (pp 907-911) Prusinkiewicz, P 2004 Modeling plant growth and development Current Opinion in Plant Biology, 7(1), (79-83) Roberts, E H., and Ellis, R H 1989 Water and Seed Survival Annals of Botany, 63(1), (pp 39-39) Saltveit ME, Morris LL 1990 Overview on chilling injury of horticultural crops In: Wang CY (ed) Chilling Injury of Horticultural Crops, (pp 3–15) Sebastian, N., Erika, H., and Christian, K 2016 Critically low soil temperatures for root growth and root morphology in three alpine plant species Alpine Botany, 126(1), (11-21) 44 h Staub J.E., Wehner T.C 1996 Temperature stress In: Zitter T.A., Hopkins D.L., Thomas C.E (Eds.), Compendium of cucurbit diseases, (pp 66-67) Verkleij, A J., de Maagd, R., Leunissen-Bijvelt, J., and de Kruijff, B 1982 Divalent cations and chlorpromazine can induce non-bilayer structures in phosphatidic acid-containing model membranes Biochimica et Biophysica Acta (BBA) Biomembranes, 684(2), (pp 255-262) Vieira, R., Tekrony, D M., Egli, D B., and Rucker, M 2001 Electrical conductivity of soybean seeds after storage in several environments Seed Science and Technology, 29(3), (pp 599-608) Villa-Hernández, J M., Dinkova, T D., Aguilar-Caballero, R., Rivera-Cabrera, F., Sánchez de Jiménez, E., and Pérez-Flores, L J 2013 Regulation of ribosome biogenesis in maize embryonic axes during germination Biochimie, 95(10), (pp 1871-1879) Woodstock, W, L., and Pollock, M., B 1965 Physiological Predetermination: Imbibition, Respiration, and Growth of Lima Bean Seeds, 150(3699), (pp 1031-1032) W., S E 1974 Phospholipids and plant membrane permeability New Phytologist, 73(3), (pp 377-420) Walters, C., Wheeler, L., and Stanwood, P C 2004 Longevity of cryogenically stored seeds Cryobiology, 48(3), 229-244 Wang, T.-M., Leskovar, D., and Cobb, B G 2014 Respiration during germination of diploid and triploid watermelon Seed Science and Technology, 42(3), (pp 313321) 45 h Wehner T.C 1981 Screening for low-temperature germination ability in cucumber Horticultural Science 16(3), (pp 399-399) Wehner T.C 1982 Genetic variation for low-temperature germination ability in cucumber Cucurbit Genet Coop, 5, (pp 16-17) Weitbrecht, K., Müller, K., and Leubner-Metzger, G 2011 First off the mark: early seed germination Journal of Experimental Botany, 62(10), (pp 3289-3309) Xiaomei, Y., Aihua, L., and Weiqi, L 2015 How membranes organize during seed germination: three patterns of dynamic lipid remodelling define chilling resistance and affect plastid biogenesis Plant, Cell & Environment, 38(7), (pp 1391-1403) Yang, H S., Dobermann, A., Lindquist, J L., Walters, D T., Arkebauer, T J., and Cassman, K G 2004 Hybrid-maize—a maize simulation model that combines two crop modeling approaches Field Crops Research, 87(2), (pp 131-154) Zhang, X D., Wang, R P., Zhang, F J., Tao, F Q., and Li, W Q 2013 Lipid profiling and tolerance to low-temperature stress in Thellungiella salsuginea in comparison with Arabidopsis thaliana Biologia Plantarum, 57(1), (pp 149153) 46 h