以生菜Lactuca sativa含水种子为试材，通过添加外源动物泛素E3连接酶激活剂小白菊内酯(Parthenolide)，及不同降温速率的程序降温处理，研究Parthenolide对程序降温过程中生菜种子抗冷性的影响。结果表明，无论在快速(60 ℃·h-1)还是慢速(3 ℃·h-1)降温过程中，添加Parthenolide都会抑制生菜种子对低温的耐受性，而且发芽率明显比对照组低，这种降低程度在降温至-20 ℃时最为明显，说明在程序降温过程中，泛素化调控通路参与种子对冷冻的响应。通过实时定量荧光PCR对泛素化相关基因和ICE1基因的表达分析表明，Parthenolide抑制了慢速降温过程中环指型E3连接酶COP1和抗冻因子ICE1的mRNA表达水平。Parthenolide对SOD活性的影响进一步证明其影响种子在程序降温过程中的耐冻性机制。
In order to study the effects of Parthenolide on chilling resistance of Lactuca sativa seeds during programmed cooling, the seeds were imbibed by exogenous animal ubiquitin E3 ligase activator Parthenolide before treated with programmed cooling with different cooling rates. The results showed that the addition of Parthenolide reduced the freezing tolerance of lettuce seeds in both fast and slow cooling treatments, and the germination rates after both cooling treatments were significantly lower than control, the effects of Parthenolide were the most obvious when the temperature was reduced to -20 ℃, which indicating that during the programmed cooling, the ubiquitination regulatory pathway did participate in the resistance to freezing in seeds. The expression of ubiquitination-related genes and ICE1 gene were detected by real-time quantitative PCR. It was found that Parthenolide inhibited the expression of RING-finger E3 ligase COP1 and anti-freezing factor ICE1 during the slow cooling. The effects of Parthenolide on the activities of SOD were further proved that during the programmed cooling process, Parthenolide may destroy the freezing resistance system of lettuce seeds
[1] 张志良,瞿伟菁. 植物生理学实验指导[M]. 北京: 高等教育出版社, 2003: 208—210.
[2] Roberts E H. Predicting the storage life of seeds[J]. Seed Science & Technology, 1973,1: 499—514.
[3] Ellis R H, Hong T D, Roberts E H. An intermediate category of seed storage behavior[J]. Journal of Experimental Botany, 1990,41: 1167—1174.
[4] 顾琦琚,韩颖颖, Ganesh,刘宝林. 罗莎绿种子的生长与低温保存研究[C]//上海市制冷学会2013年学术年会论文集. 上海: 上海市制冷学会, 2013: 4.
[5] Li C J, Guo S F, Shi T M. Culture supernatants of breast cancer cell line MDA-MB-231 treated with parthenolide inhibit the proliferation, migration, and lumen formation capacity of human umbilical vein endothelial cells[J]. Chinese Medical Journal(English edition), 2012,125(12): 2195—2199.
[6] Gopal Y N, Chanehorn E, Van-Dyke M W. Parthenolide promotes the ubiquitination of MDM2 and activates p53 cellular functions[J]. Molecular Cancer Therapeutics, 2009,8(3): 552—562.
[7] 任伟嘉. 两种玫瑰品种的抗寒性研究[D]. 哈尔滨: 东北林业大学硕士学位论文, 2013.
[8] Wang S M, Wan C G, Wang Y R. The characteristics of K+, Na+ and free praline distribution in several drought resistant plants of the Alxa desert, China[J]. Journal of Arid Environments, 2004,56: 525—539.
[9] Hershko A, Ciechanover A. The ubiquitin system[J]. Annual Review Biochemistry, 1998,67: 425—479.
[10] 王鹏. RING结构泛素连接酶RNF186的功能研究[D]. 合肥: 中国科学技术大学博士学位论文, 2013.
[11] Dong C H, Agarwal M, Zhang Y, Xie Q, Zhu J K. The negative regulator of plant cold responses, HOS1, is a RING E3 ligase that mediates the ubiquitination and degradation of ICE1[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006,103(21): 8281—8286.
[12] Lee H, Xiong L, Gong Z, Ishitani M, Stevenson B, Zhu J K. The Arabidopsis HOS1 gene negatively regulates cold signal transduction and encodes a RING finger protein that displays cold-regulated nucleo-cytoplasmic partitioning[J]. Genes & Development, 2001,15(7): 912—924.
[13] 杨家森,张洪涛,李新国,毕玉平. 拟南芥CBF冷反应通路[J]. 植物生理学通讯, 2006(1): 155—161.
[14] Fryer M J, Andrews J R, Oxborough K. Relationship between CO2 assimilation, photosynthetic electron transport, and active O2 metabolism in leaves of maize in the field during periods of low temperature[J]. Plant Physiology, 1998,116: 571—580.
[15] Prasad T K. Role of catalase in inducing chilling tolerance in preemergent maize seedlings[J]. Plant Physiology, 1997,114: 1369—1376.
[16] Keefe P D, Moore K G. Freeze desiccation: a second mechanism for the survival of hydrated lettuce (Lactuca sativa L.) seed at sub-zero temperatures[J]. Annals of Botany, 1981,47(5): 635—645.
[17] 曾韶西,王以柔. 低温胁迫对黄瓜子叶抗坏血酸过氧化物酶活性和谷胱甘肽含量的影响[J]. 植物生理学报, 1990,16(1): 37—42.
[18] 陈杰忠,徐春香,梁立峰. 低温对香蕉叶片中蛋白质及脯氨酸的影响[J]. 华南农业大学学报, 1999,20(3): 54—58.
[19] Dlauney A J, Verma D P S. Proline biosynthesis and osmoregulation in plants[J]. Plant Journal, 1993(4): 215—223.
[20] Khedr A H, Abbas M A, Wahid A A. Proline induces the expression of salt-stress -responsive proteins and may improve the adaptation of Pancratium maritimum L. to salt-stress[J]. Journal of Experimental Botany, 2003,54: 2553—2562.
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