外源Ca~(2+)提高百合耐热性的生理机制初探
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摘要
为探讨Ca~(2+)对百合耐热性的影响及其生理机制,以东方百合杂种系西伯利亚为试验材料,研究CaCl_2、钙离子螯合剂EGTA、钙调蛋白抑制剂三氟拉嗪(TFP)处理对40℃高温胁迫下百合热害指数、抗性相关生理指标和相关热响应基因表达的影响。结果表明,高温胁迫下,15 mmol/L CaCl_2处理能够减缓膜透性的加大、叶绿素的降解和丙二醛的积累,提高脯氨酸、可溶性蛋白含量和超氧化物歧化酶活性,促进热响应基因LoCaM3和LoHsfA3a的表达,从而提高植株的耐热性,降低热害指数。10 mmol/L EGTA和200μmol/L TFP的作用则相反。表明作为植物生长所必需的大量元素,Ca~(2+)也可能通过Ca~(2+)-CaM信使系统提高渗透调节能力、抗氧化能力、光合能力和蛋白质活性,以维持细胞稳态来调节百合对高温逆境的适应性。
To determine the effect and physiological mechanism of calcium on the thermotolerance of lily, the heat injury index,correlative physiological indexes and heat-responsed genes under high temperature stress of 40 ℃ were investigated by treating Lilium oriental Siberia with exogenous Ca~(2+), Ca~(2+) chelating agent EGTA and calmodulin antagonist TFP.The results showed that under the circumstance of heatstress,15 mmol/L Ca~(2+) treatment alleviated leakage of electrolyte, degradation of chlorophyll and the accumulation of MDA caused by heat stress,enhanced the content of proline,soluble protein and activity of SOD,and induced the expression of heat-responsed genes LoCaM3 and LoHsfA3a,which increased tolerance to heat stress and decreased heat injury index.The role of 10 mmol/L EGTA or 200 μmol/L TFP was opposite. The results indicated that as one essential macroelement towards plant growth,Ca~(2+) might regulate the heat resistance of lily through Ca~(2+)-CaM signal transduction by improving the ability of osmoregulation, antioxidation, photosynthesis and the activity of protein to maintain cellular homeostasis.
To determine the effect and physiological mechanism of calcium on the thermotolerance of lily, the heat injury index,correlative physiological indexes and heat-responsed genes under high temperature stress of 40 ℃ were investigated by treating Lilium oriental Siberia with exogenous Ca~(2+), Ca~(2+) chelating agent EGTA and calmodulin antagonist TFP.The results showed that under the circumstance of heatstress,15 mmol/L Ca~(2+) treatment alleviated leakage of electrolyte, degradation of chlorophyll and the accumulation of MDA caused by heat stress,enhanced the content of proline,soluble protein and activity of SOD,and induced the expression of heat-responsed genes LoCaM3 and LoHsfA3a,which increased tolerance to heat stress and decreased heat injury index.The role of 10 mmol/L EGTA or 200 μmol/L TFP was opposite. The results indicated that as one essential macroelement towards plant growth,Ca~(2+) might regulate the heat resistance of lily through Ca~(2+)-CaM signal transduction by improving the ability of osmoregulation, antioxidation, photosynthesis and the activity of protein to maintain cellular homeostasis.
引文
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[11] 吴在生,曹晓娟,张智,等.高温胁迫下东方百合‘Sorbonne’ 形态及生理响应[J].西北农业学报,2011,20(2):174-177.
[12] 杨炜茹,张启翔,孙明,等.高温胁迫对岷江百合幼苗耐热指数和理化特性的影响[J].华南农业大学学报,2010,31(1):51-54.
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[14] YIN H,CHEN Q M,YI M F.Effects of short-term heat stress on oxidative damage and responses of antioxidant system in Lilium longiflorum[J].Plant Growth Regulation,2008,54(1):45-53.
[15] 刘雪凝,杨利平,马川,等.温汤处理种球对亚洲百合幼苗耐热性的影响[J].中国农业科学,2010,43(6):1314-1320.
[16] 刘雪凝,杨利平.热激锻炼对亚洲百合耐热性的诱导[J].东北林业大学学报,2011,39(5):64-66.
[17] 向地英,张钢,杨利平.热激诱导对高温胁迫下百合相对电导率及电阻抗参数的影响[J].西北农业学报,2014,23(9):189-195.
[18] 陈秋明,尹慧,李晓艳,等.高温胁迫下外源水杨酸对百合抗氧化系统的影响[J].中国农业大学学报,2008,13(2):44-48.
[19] 李利红,杨亚军,赵会杰,等.外源Ca2+对高温强光胁迫下灌浆期小麦叶片光合机构运转的影响[J].植物生理学通讯,2009,45(9):851-854.
[20] 杨卫星,黑刚刚,李姣姣,等.外源Ca2+对高温胁迫下半夏光合参数及有效成分积累的影响[J].中国中药杂志,2014,39(14):2614-2618.
[21] 陈芳芳,郝福玲,汪天,等.Ca2+处理对不同藤本月季品种的耐热性影响[J].安徽农业大学学报,2013,40(3):400-405.
[22] 赵冰,付玉梅,丁惠惠,等.Ca2+处理对秦岭高山杜鹃耐热性的影响[J].西北林学院学报,2010,25(6):29-32.
[23] 陈贵林,贾开志.钙和钙调素拮抗剂对高温胁迫下茄子幼苗抗氧化系统的影响[J].中国农业科学,2005,38(1):197-202.
[24] 熊瑛,吕强,刘素云,等.ABA及Ca2+/CaM信使系统对高温逆境下棉苗根系生长的调控[J].干旱地区农业研究,2010,28(6):165-169.
[25] 程千钉,刘桂芳,杨利平.百合新种质苗期耐热性比较[J].河北农业大学学报,2012,35(5):34-39.
[26] 李蕊,杨利平,刘雪凝.百合杂交种及亲本耐热性比较[J].中国农业科学,2011,44(6):1201-1209.
[27] 崔光芬,王祥宁,蒋亚莲,等.OT百合新品种‘金门’[J].园艺学报,2014,41(7):1517-1518.
[28] 冯丽媛,王文和,赵祥云,等.景观应用百合新品种‘云景红’[J].园艺学报,2014,41(6):1275-1276.
[29] 尚爱芹,高永鹤,段龙飞,等.逆境诱导转录因子AtDREB2A转化百合的研究[J].园艺学报,2014,41(1):149-156.
[30] GONG B H,YI J,WU J,et al.LlHSFA1,a novel heat stress transcription factor in lily(Lilium longiflorum), can interact with LlHSFA2 and enhance the thermotolerance of transgenic Arabidopsis thaliana[J].Plant Cell Reports,2014,33(9):1519-1533.
[31] XIN H B,ZHANG H,CHEN L,et al.Cloning and characterization of HsfA2 from lily(Lilium longiflorum)[J].Plant Cell Reports,2010,29(8):875-885.
[32] XIN H B,ZHANG H,ZHONG X H,et al.Over-expression of LlHsfA2b,a lily heat shock transcription factor lacking trans-activation activity in yeast,can enhance tolerance to heat and oxidative stress in transgenic arabidopsis seedlings[J].Plant Cell Tissue and Organ Culture,2017,130(3):617-629.
[33] 宫本贺.百合热激转录因子L1HSFA1及其下游热激蛋白L1HSP70响应热胁迫的机制解析[D].北京:中国农业大学,2014.
[34] MU C J,ZHANG S J,YU G Z,et al.Overexpression off small heat shock protein LimHSP16.45 in Arabidopsis enhances tolerance to abiotic stresses[J].PLoS ONE,2013,8(12):e82264.
[35] WANG W,VINOCUR B,SHOSEYOV O,et al.Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response[J].Trends in Plant Science,2004,9(5):244-252.
[2] HASHIMOTO K,KUDLA J.Calcium decoding mechanisms in plants[J].Biochimie,2011,93:2054-2059.
[3] 秦舒浩,李玲玲,陈娜娜,等.外源Ca2+对高温强光下西葫芦幼苗形态特征、光合特性及荧光参数的影响[J].应用生态学报,2010,21(11):2830-2835.
[4] 宰学明,夏连全,闫道良,等.外源Ca2+对高温强光胁迫下滨梅幼苗的保护效应[J].西北植物学报,2011,31(3):558-563.
[5] LIU H T,LI B,SHAN Z L,et al.Calmodulin is involved in heat shock signal transduction in wheat[J].Plant Physiology,2003,132:1186-1195.
[6] 于守超, 张秀省, 冀芦莎.基于Photoshop CS5的植物叶面积测定方法[J].湖北农业科学,2012,51(15):3340-3342.
[7] 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000.
[8] SCHMITTGEN T D,LIVAK K J.Analyzing real-time PCR data by the comparative CT method[J].Nature Protocols,2008,3(6):1101-1108.
[9] CAO X,YI J,WU Z,et al.Involvement of Ca2+ and CaM3 in regulation of thermotolerance in lily(Lilium longiflorum)[J]. Plant Molecular Biology Reporter,2013,31(6):1293-1304.
[10] WU Z,LIANG J H,WANG C P,et al.Overexpression of two novel HsfA3s from lily in Arabidopsis confer increased thermotolerance and salt sensitivity via alterations in proline catabolism[J].Journal of Experimental Botany,2018,69(8):2005-2021.
[11] 吴在生,曹晓娟,张智,等.高温胁迫下东方百合‘Sorbonne’ 形态及生理响应[J].西北农业学报,2011,20(2):174-177.
[12] 杨炜茹,张启翔,孙明,等.高温胁迫对岷江百合幼苗耐热指数和理化特性的影响[J].华南农业大学学报,2010,31(1):51-54.
[13] 周斯建,义鸣放,穆鼎.高温胁迫下铁炮百合幼苗形态及生理反应的初步研究[J].园艺学报,2005,32(1):145-147.
[14] YIN H,CHEN Q M,YI M F.Effects of short-term heat stress on oxidative damage and responses of antioxidant system in Lilium longiflorum[J].Plant Growth Regulation,2008,54(1):45-53.
[15] 刘雪凝,杨利平,马川,等.温汤处理种球对亚洲百合幼苗耐热性的影响[J].中国农业科学,2010,43(6):1314-1320.
[16] 刘雪凝,杨利平.热激锻炼对亚洲百合耐热性的诱导[J].东北林业大学学报,2011,39(5):64-66.
[17] 向地英,张钢,杨利平.热激诱导对高温胁迫下百合相对电导率及电阻抗参数的影响[J].西北农业学报,2014,23(9):189-195.
[18] 陈秋明,尹慧,李晓艳,等.高温胁迫下外源水杨酸对百合抗氧化系统的影响[J].中国农业大学学报,2008,13(2):44-48.
[19] 李利红,杨亚军,赵会杰,等.外源Ca2+对高温强光胁迫下灌浆期小麦叶片光合机构运转的影响[J].植物生理学通讯,2009,45(9):851-854.
[20] 杨卫星,黑刚刚,李姣姣,等.外源Ca2+对高温胁迫下半夏光合参数及有效成分积累的影响[J].中国中药杂志,2014,39(14):2614-2618.
[21] 陈芳芳,郝福玲,汪天,等.Ca2+处理对不同藤本月季品种的耐热性影响[J].安徽农业大学学报,2013,40(3):400-405.
[22] 赵冰,付玉梅,丁惠惠,等.Ca2+处理对秦岭高山杜鹃耐热性的影响[J].西北林学院学报,2010,25(6):29-32.
[23] 陈贵林,贾开志.钙和钙调素拮抗剂对高温胁迫下茄子幼苗抗氧化系统的影响[J].中国农业科学,2005,38(1):197-202.
[24] 熊瑛,吕强,刘素云,等.ABA及Ca2+/CaM信使系统对高温逆境下棉苗根系生长的调控[J].干旱地区农业研究,2010,28(6):165-169.
[25] 程千钉,刘桂芳,杨利平.百合新种质苗期耐热性比较[J].河北农业大学学报,2012,35(5):34-39.
[26] 李蕊,杨利平,刘雪凝.百合杂交种及亲本耐热性比较[J].中国农业科学,2011,44(6):1201-1209.
[27] 崔光芬,王祥宁,蒋亚莲,等.OT百合新品种‘金门’[J].园艺学报,2014,41(7):1517-1518.
[28] 冯丽媛,王文和,赵祥云,等.景观应用百合新品种‘云景红’[J].园艺学报,2014,41(6):1275-1276.
[29] 尚爱芹,高永鹤,段龙飞,等.逆境诱导转录因子AtDREB2A转化百合的研究[J].园艺学报,2014,41(1):149-156.
[30] GONG B H,YI J,WU J,et al.LlHSFA1,a novel heat stress transcription factor in lily(Lilium longiflorum), can interact with LlHSFA2 and enhance the thermotolerance of transgenic Arabidopsis thaliana[J].Plant Cell Reports,2014,33(9):1519-1533.
[31] XIN H B,ZHANG H,CHEN L,et al.Cloning and characterization of HsfA2 from lily(Lilium longiflorum)[J].Plant Cell Reports,2010,29(8):875-885.
[32] XIN H B,ZHANG H,ZHONG X H,et al.Over-expression of LlHsfA2b,a lily heat shock transcription factor lacking trans-activation activity in yeast,can enhance tolerance to heat and oxidative stress in transgenic arabidopsis seedlings[J].Plant Cell Tissue and Organ Culture,2017,130(3):617-629.
[33] 宫本贺.百合热激转录因子L1HSFA1及其下游热激蛋白L1HSP70响应热胁迫的机制解析[D].北京:中国农业大学,2014.
[34] MU C J,ZHANG S J,YU G Z,et al.Overexpression off small heat shock protein LimHSP16.45 in Arabidopsis enhances tolerance to abiotic stresses[J].PLoS ONE,2013,8(12):e82264.
[35] WANG W,VINOCUR B,SHOSEYOV O,et al.Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response[J].Trends in Plant Science,2004,9(5):244-252.