温度和盐胁迫对草莓开花进程和相关基因表达的影响
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摘要
为探究草莓花器官对高温、低温、盐胁迫的响应特征,该研究以草莓品种‘甜查理’为试材,设置0℃、4℃、25℃(CK)、37℃、200 mmol/L NaCl、400 mmol/L NaCl等逆境胁迫处理,考察草莓花外观形态、生长发育状态、花器官抗氧化酶活性,以及3个草莓花器官分生组织特异性基因、12个成花途径基因、3个抗氧化酶基因、6个抗逆基因的表达等。结果显示:(1)0℃、4℃、37℃、200 mmol/L NaCl、400 mmol/L NaCl溶液处理均抑制了草莓花的开放,0℃低温显著抑制草莓花器官的开放,并对草莓的花器官造成伤害。(2)当草莓植株受到高温、低温、盐胁迫时,其花器官内抗氧化酶SOD、POD、CAT活性及MDA含量均得到提高,其相关抗氧化酶基因的表达水平也相应提高。(3)0℃、4℃、37℃以及200 mmol/L NaCl、400 mmol/L NaCl溶液处理均显著降低了草莓花器官FaAP1、FaLFY、FaFD、FaSOC1等成花基因的表达量;0℃低温诱导了FaICE、FaRD29A、FaCOR、FaAFP等4个抗寒基因的表达水平;200 mmol/L NaCl、400 mmol/L NaCl的盐胁迫显著提高了FaP5CS基因的表达量。研究表明,0℃、4℃、37℃、200 mmol/L NaCl、400 mmol/L NaCl溶液处理不同程度抑制了草莓的开花进程,对草莓花器官造成伤害;不同逆境胁迫处理显著提高了草莓花器官SOD、POD、CAT活性以及MDA含量,相应的抗氧化酶基因表达得到上调,同时显著降低了草莓成花途径相关基因表达,增强了相关抗逆基因的表达水平。
In order to explore the response characteristics of strawberry flower organs to high temperature, low temperature and salt stress, we used strawberry variety ‘Sweet Charlie' as the test material. 0 ℃, 4 ℃, 25 ℃(CK), 37 ℃, 200 mmol/L NaCl, 400 mmol/L NaCl and other stress treatments were used to investigate the appearance, growth and development of flowering organs, antioxidant enzyme activities of flower organs, and the expression of three strawberry flower organ meristem genes, 12 genes of flowering pathways, and three antioxidants enzyme genes and six stress-resistant genes. The results showed that:(1) 0 ℃, 4 ℃, 37 ℃, 200 mmol/L NaCl, 400 mmol/L NaCl treatment inhibited the opening of strawberry flowers, 0 ℃ low temperature significantly inhibited the opening of strawberry flower organs, and caused the flower organs of strawberries hurt.(2) When strawberry plants were exposed to high temperature, low temperature and salt stress, the activities of antioxidant enzymes SOD, POD, CAT and MDA content in flower organs were increased, and the expression levels of related antioxidant enzyme genes were also increased.(3) 0 ℃, 4 ℃, 37 ℃ and 200 mmol/L NaCl, 400 mmol/L NaCl solution significantly reduced the expression of FaAP1, FaLFY, FaFD, FaSOC1 and other flowering genes in strawberry flower organs; 0 ℃ low temperature induced the expression levels of four cold-resistant genes such as FaICE, FaRD29A, FaCOR and FaAFP; salt stress of 200 mmol/L NaCl and 400 mmol/L NaCl significantly increased the expression of FaP5CS gene. Studies showed that 0 ℃, 4 ℃, 37 ℃, 200 mmol/L NaCl, 400 mmol/L NaCl solution inhibited the flowering process of strawberry and caused damage to strawberry flower organs. Different stress treatment significantly increased the SOD of strawberry flower organs. The activities of POD, CAT and MDA content, the expression of the corresponding antioxidant enzyme genes were up-regulated, and the expression of genes involved in the flowering pathway of strawberry was significantly decreased, and the expression level of related stress-resistant genes was enhanced.
In order to explore the response characteristics of strawberry flower organs to high temperature, low temperature and salt stress, we used strawberry variety ‘Sweet Charlie' as the test material. 0 ℃, 4 ℃, 25 ℃(CK), 37 ℃, 200 mmol/L NaCl, 400 mmol/L NaCl and other stress treatments were used to investigate the appearance, growth and development of flowering organs, antioxidant enzyme activities of flower organs, and the expression of three strawberry flower organ meristem genes, 12 genes of flowering pathways, and three antioxidants enzyme genes and six stress-resistant genes. The results showed that:(1) 0 ℃, 4 ℃, 37 ℃, 200 mmol/L NaCl, 400 mmol/L NaCl treatment inhibited the opening of strawberry flowers, 0 ℃ low temperature significantly inhibited the opening of strawberry flower organs, and caused the flower organs of strawberries hurt.(2) When strawberry plants were exposed to high temperature, low temperature and salt stress, the activities of antioxidant enzymes SOD, POD, CAT and MDA content in flower organs were increased, and the expression levels of related antioxidant enzyme genes were also increased.(3) 0 ℃, 4 ℃, 37 ℃ and 200 mmol/L NaCl, 400 mmol/L NaCl solution significantly reduced the expression of FaAP1, FaLFY, FaFD, FaSOC1 and other flowering genes in strawberry flower organs; 0 ℃ low temperature induced the expression levels of four cold-resistant genes such as FaICE, FaRD29A, FaCOR and FaAFP; salt stress of 200 mmol/L NaCl and 400 mmol/L NaCl significantly increased the expression of FaP5CS gene. Studies showed that 0 ℃, 4 ℃, 37 ℃, 200 mmol/L NaCl, 400 mmol/L NaCl solution inhibited the flowering process of strawberry and caused damage to strawberry flower organs. Different stress treatment significantly increased the SOD of strawberry flower organs. The activities of POD, CAT and MDA content, the expression of the corresponding antioxidant enzyme genes were up-regulated, and the expression of genes involved in the flowering pathway of strawberry was significantly decreased, and the expression level of related stress-resistant genes was enhanced.
引文
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[2] 刘贝贝,陈利娜,李好先,等.果树响应低温胁迫的分子机制及抗寒性鉴定方法研究进展[J].北方园艺,2017,(15):174-179.LIU B B,CHEN L N,LI H X,et al.Advances in molecular mechanisms and identification methods of cold resistance in response to low temperature stress[J].Northern Gardening,2017,(15):174-179.
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[8] 张超博,李傲,崔梦杰,等.干旱胁迫下‘宁玉’草莓生长发育及相关基因表达特点[J].园艺学报,2017,(12):2 351-2 360.ZHANG C B,LI A,CUI M J,et al.Growth and development of related genes in ‘Ningyu’ under drought stress[J].Journal of Horticulture,2017,(12):2 351-2 360.
[9] STRIK B C.Flower bud initiation in strawberry cultivars[J].Fruit Varieties Journal,1985,39(1):5.
[10] LEDESMA N A,NAKATA M,SUGIYAMA N.Effect of high temperature stress on the reproductive growth of strawberry cvs.‘Nyoho’ and ‘Toyonoka’[J].Scientia Horticulturae,2008,116(2):186-193.
[11] 李青云.草莓对盐胁迫的反应及调控研究[D].河北保定:河北农业大学,2005.
[12] LI K ,WANG Y,HAN C,et al.GA signaling and CO/FTregulatory module mediate salt-induced late flowering in Arabidopsis thaliana[J].Plant Growth Regulation,2007,53(3):195-206.
[13] 梁政杰,田芳,王晓云,等.早熟草莓品种甜查理优质高产栽培技术[J].农业科技通讯,2017,(3):254-255.LIANG Z J,TIAN F,WANG X Y,et al.Early-ripening strawberry variety sweet Charlie high-qualit and high-yield cultivation techniques[J].Agricultural Technology Communication,2017,(3):254-255.
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[17] 陈毓荃.生物化学实验方法和技术[M].北京:科学出版社,2000.
[18] BAIER M,NOCTOR G,FOVER C H,et al.Antisense suppression of 2-cysteine peroxiredoxin in arabidopsis specifically enhances the activities and expression of enzymes associated with ascorbate metabolism but not glutathione metabolism[J].Plant Physiology,2000,124(2):823-832.
[19] MILLER G,MITTLER R.Double mutants deficient in cytosolic and thylakoid ascorbate peroxidase reveal a complex mode of interaction between reactive oxygen species,plant development,and response to abiotic stresses[J].Plant Physiology,2007,144(4):1 777-1 785.
[20] SU Z,MA X,GUO H,et al.Flower development under drought stress:morphological and transcriptomic analyses reveal acute responses and long-term acclimation in Arabidopsis[J].Plant Cell,2013,25(10):3 785-3 807.
[21] SEO E,LEE H,JIN J,et al.Crosstalk between cold response and flowering in arabidopsis is mediated through the flowering-time gene SOC1 and its upstream negative regulator FLC[J].Plant Cell,2009,21(10):3 185.
[22] FRANKLIN K A,WHITELAM G C.Light-quality regulation of freezing tolerance in Arabidopsis thaliana[J].Nature Genetics,2007,39(11):1 410-1 413.
[23] BBARNABAS B,JAGER K,FEHER A.The effect of drought and heat stress on reproductive processes in cereals.[J].Plant Cell & Environment,2010,31(1):11-38.
[24] BALASUBRAMANIAN S,SURESHKUMAR S,et al.Potent induction of Arabidopsis thaliana flowering by elevated growth temperature[J].Plos Genetics,2006,2(7):e106.
[25] ZINN K E,TUNCOZDEMIR M,HARPERr J F.Temperature stress and plant sexual reproduction:uncovering the weakest links[J].Journal of Experimental Botany,2010,61(7):1 959-1 968.
[26] 管志勇,陈发棣,陈素梅,等.NaCl胁迫对菊花花序形态及生理指标的影响[J].西北植物学报,2009,29(8):1 624-1 629.GUAN Z Y,CHEN F D,CHEN S M,et al.Effects of NaCl stress on the morphology and physiological indexes of chrysanthemum inflorescence[J].Acta Botanica Boreali-Occidentalia Sinica,2009,29(8):1 624-1 629.
[27] KIM H J,HYUN Y,PARK J Y,et al.A genetic link between cold responses and flowering time through FVE in Arabidopsis thaliana[J].Nature Genetics,2004,36(2):167.
[28] BLAZQUEZ M A,AHN J H,WEIGEL D.A thermosensory pathway controlling flowering time in Arabidopsis thaliana[J].Nature Genetics,2003,33(2):168.
[29] BALASUBRAMANIAN S,SURESHKUMAR S,LEMPE J,et al.Potent induction of Arabidopsis thaliana flowering by elevated growth temperature[J].Plos Genetics,2006,2(7):e106.
[30] YOSHIHIRO N,YOHEI H,KATSUHIKO S,et al.Flowering retardation by high temperature in chrysanthemums:involvement of FLOWERING LOCUS T-like 3 gene repression[J].Journal of Experimental Botany,2013,64(4):909-920.
[31] PNUELI L,LIANG H,ROZENBERG M,et al.Growth suppression,altered stomatal responses,and augmented induction of heat shock proteins in cytosolic ascorbate peroxidase (Apx1)-deficient Arabidopsis plants[J].Plant Journal,2003,34(2):187-203.
[32] 杨春祥,李宪利,高东升,等低温胁迫对油桃花器官膜脂过氧化和保护酶活性的影响[J].果树学报,2005,22(1).YANG C X,LI X L,GAO D S.et al.Effects of low temperature stress on membrane lipid peroxidation and protective enzyme activities in nectarine flower organs[J].Journal of Fruit Science,2005,22(1)
[33] 马翠云,封雷,陶书田,等.低温胁迫对梨花器官及授粉受精的影响[J].南京农业大学学报,2014,37(1):53-59.MA C Y,FENG L,TAO S T,et al.Effects of low temperature stress on pear organs and pollination and fertilization[J].Journal of Nanjing Agricultural University,2014,37(1):53-59.
[34] KAMIAB F,TALAIE A,KHEZRI M,et al.Exogenous application of free polyamines enhance salt tolerance of pistachio (Pistacia vera L.) seedlings[J].Plant Growth Regulation,2014,72(3):257-268.
[35] 朱婷婷,王彦霞,裴丽丽,等.植物蛋白激酶与作物非生物胁迫抗性的研究[J].植物遗传资源学报,2017,18(4):763-770.ZHU T T,WANG Y X,PEI L L,et al.Study on plant protein kinase and resistance to abiotic stress in crops[J].Journal of Plant Genetic Resources,2017,18(4):763-770.
[36] 黄永会,刘永翔,朱英,等.COR基因在植物抗寒基因工程中的作用[J].贵州农业科学,2014,(12):37-42.HUANG Y H,LIU Y X,ZHU Y,et al.The role of COR gene in plant cold resistance genetic engineering[J].Guizhou Agricultural Science,2014,(12):37-42.
[37] THOMASHOW M F.Role of cold-responsive genes in plant freezing tolerance[J].Plant Physiology,1998,118(1):1-7.
[2] 刘贝贝,陈利娜,李好先,等.果树响应低温胁迫的分子机制及抗寒性鉴定方法研究进展[J].北方园艺,2017,(15):174-179.LIU B B,CHEN L N,LI H X,et al.Advances in molecular mechanisms and identification methods of cold resistance in response to low temperature stress[J].Northern Gardening,2017,(15):174-179.
[3] 任军,黄志霖,曾立雄,等.低温胁迫下植物生理反应机理研究进展[J].世界林业研究,2013,26(6):15-20.REN J,HUANG Z L,ZENG L X,et al.Advances in research on mechanism of plant physiological response under low temperature stress[J].World Forestry Research,2013,26(6):15-20.
[4] VINOCUR B,ALTMAN A.Recent advances in engineering plant tolerance to abiotic stress:achievements and limitations[J].Curr.Opin.Biotechnol,2005,16(2):123-132.
[5] MITTAL D,MADHYASTHA D A,Grover A.Gene expression analysis in response to low and high temperature and oxidative stresses in rice:combination of stresses evokes different transcriptional changes as against stresses applied individually[J].Plant Science,2012,197(4):102-113.
[6] 何晓明,林毓娥,陈清华,等.高温对黄瓜幼苗生长、脯氨酸含量及SOD酶活性的影响[J].上海交通大学学报(农业科学版),2002,20(1):30-33.HE X M,LIN S E,CHEN Q H,et al.Effects of high temperature on growth,proline content and SOD activity of cucumber seedlings[J].Journal of Shanghai Jiaotong University (Agricultural Science Edition),2002,20(1):30-33.
[7] 黄建昌.草莓对高温胁迫的有关生理反应[J].仲恺农业工程学院学报,1993,6(1):34-38.HUANG J C.Physiological responses of strawberry to high temperature stress[J].Journal of Zhongkai University of Agriculture and Engineering,1993,6(1):34-38.
[8] 张超博,李傲,崔梦杰,等.干旱胁迫下‘宁玉’草莓生长发育及相关基因表达特点[J].园艺学报,2017,(12):2 351-2 360.ZHANG C B,LI A,CUI M J,et al.Growth and development of related genes in ‘Ningyu’ under drought stress[J].Journal of Horticulture,2017,(12):2 351-2 360.
[9] STRIK B C.Flower bud initiation in strawberry cultivars[J].Fruit Varieties Journal,1985,39(1):5.
[10] LEDESMA N A,NAKATA M,SUGIYAMA N.Effect of high temperature stress on the reproductive growth of strawberry cvs.‘Nyoho’ and ‘Toyonoka’[J].Scientia Horticulturae,2008,116(2):186-193.
[11] 李青云.草莓对盐胁迫的反应及调控研究[D].河北保定:河北农业大学,2005.
[12] LI K ,WANG Y,HAN C,et al.GA signaling and CO/FTregulatory module mediate salt-induced late flowering in Arabidopsis thaliana[J].Plant Growth Regulation,2007,53(3):195-206.
[13] 梁政杰,田芳,王晓云,等.早熟草莓品种甜查理优质高产栽培技术[J].农业科技通讯,2017,(3):254-255.LIANG Z J,TIAN F,WANG X Y,et al.Early-ripening strawberry variety sweet Charlie high-qualit and high-yield cultivation techniques[J].Agricultural Technology Communication,2017,(3):254-255.
[14] DARNELL R L,CANTLIFFE D J,KIRSCHBAUM D S ,et al.The Physiology of Flowering in Strawberry[M]// Horticultural Reviews,Volume 28.John Wiley & Sons,Inc.2003.
[15] 李合生.植物生理生化试验原理和技术[M].北京:高等教育出版社,2000.
[16] 李璇,王升,岳红,等.不同 pH 值和酶提取体系对丹参抗氧化酶活性的影响[J].中国现代中药,2011,13(12):46-49.LI X,WANG S,YUE H,et al.Effects of different pH values and enzyme extraction systems on the activities of antioxidant enzymes in Salvia miltiorrhiza[J].Chinese Modern Chinese Medicine,2011,13(12):46-49.
[17] 陈毓荃.生物化学实验方法和技术[M].北京:科学出版社,2000.
[18] BAIER M,NOCTOR G,FOVER C H,et al.Antisense suppression of 2-cysteine peroxiredoxin in arabidopsis specifically enhances the activities and expression of enzymes associated with ascorbate metabolism but not glutathione metabolism[J].Plant Physiology,2000,124(2):823-832.
[19] MILLER G,MITTLER R.Double mutants deficient in cytosolic and thylakoid ascorbate peroxidase reveal a complex mode of interaction between reactive oxygen species,plant development,and response to abiotic stresses[J].Plant Physiology,2007,144(4):1 777-1 785.
[20] SU Z,MA X,GUO H,et al.Flower development under drought stress:morphological and transcriptomic analyses reveal acute responses and long-term acclimation in Arabidopsis[J].Plant Cell,2013,25(10):3 785-3 807.
[21] SEO E,LEE H,JIN J,et al.Crosstalk between cold response and flowering in arabidopsis is mediated through the flowering-time gene SOC1 and its upstream negative regulator FLC[J].Plant Cell,2009,21(10):3 185.
[22] FRANKLIN K A,WHITELAM G C.Light-quality regulation of freezing tolerance in Arabidopsis thaliana[J].Nature Genetics,2007,39(11):1 410-1 413.
[23] BBARNABAS B,JAGER K,FEHER A.The effect of drought and heat stress on reproductive processes in cereals.[J].Plant Cell & Environment,2010,31(1):11-38.
[24] BALASUBRAMANIAN S,SURESHKUMAR S,et al.Potent induction of Arabidopsis thaliana flowering by elevated growth temperature[J].Plos Genetics,2006,2(7):e106.
[25] ZINN K E,TUNCOZDEMIR M,HARPERr J F.Temperature stress and plant sexual reproduction:uncovering the weakest links[J].Journal of Experimental Botany,2010,61(7):1 959-1 968.
[26] 管志勇,陈发棣,陈素梅,等.NaCl胁迫对菊花花序形态及生理指标的影响[J].西北植物学报,2009,29(8):1 624-1 629.GUAN Z Y,CHEN F D,CHEN S M,et al.Effects of NaCl stress on the morphology and physiological indexes of chrysanthemum inflorescence[J].Acta Botanica Boreali-Occidentalia Sinica,2009,29(8):1 624-1 629.
[27] KIM H J,HYUN Y,PARK J Y,et al.A genetic link between cold responses and flowering time through FVE in Arabidopsis thaliana[J].Nature Genetics,2004,36(2):167.
[28] BLAZQUEZ M A,AHN J H,WEIGEL D.A thermosensory pathway controlling flowering time in Arabidopsis thaliana[J].Nature Genetics,2003,33(2):168.
[29] BALASUBRAMANIAN S,SURESHKUMAR S,LEMPE J,et al.Potent induction of Arabidopsis thaliana flowering by elevated growth temperature[J].Plos Genetics,2006,2(7):e106.
[30] YOSHIHIRO N,YOHEI H,KATSUHIKO S,et al.Flowering retardation by high temperature in chrysanthemums:involvement of FLOWERING LOCUS T-like 3 gene repression[J].Journal of Experimental Botany,2013,64(4):909-920.
[31] PNUELI L,LIANG H,ROZENBERG M,et al.Growth suppression,altered stomatal responses,and augmented induction of heat shock proteins in cytosolic ascorbate peroxidase (Apx1)-deficient Arabidopsis plants[J].Plant Journal,2003,34(2):187-203.
[32] 杨春祥,李宪利,高东升,等低温胁迫对油桃花器官膜脂过氧化和保护酶活性的影响[J].果树学报,2005,22(1).YANG C X,LI X L,GAO D S.et al.Effects of low temperature stress on membrane lipid peroxidation and protective enzyme activities in nectarine flower organs[J].Journal of Fruit Science,2005,22(1)
[33] 马翠云,封雷,陶书田,等.低温胁迫对梨花器官及授粉受精的影响[J].南京农业大学学报,2014,37(1):53-59.MA C Y,FENG L,TAO S T,et al.Effects of low temperature stress on pear organs and pollination and fertilization[J].Journal of Nanjing Agricultural University,2014,37(1):53-59.
[34] KAMIAB F,TALAIE A,KHEZRI M,et al.Exogenous application of free polyamines enhance salt tolerance of pistachio (Pistacia vera L.) seedlings[J].Plant Growth Regulation,2014,72(3):257-268.
[35] 朱婷婷,王彦霞,裴丽丽,等.植物蛋白激酶与作物非生物胁迫抗性的研究[J].植物遗传资源学报,2017,18(4):763-770.ZHU T T,WANG Y X,PEI L L,et al.Study on plant protein kinase and resistance to abiotic stress in crops[J].Journal of Plant Genetic Resources,2017,18(4):763-770.
[36] 黄永会,刘永翔,朱英,等.COR基因在植物抗寒基因工程中的作用[J].贵州农业科学,2014,(12):37-42.HUANG Y H,LIU Y X,ZHU Y,et al.The role of COR gene in plant cold resistance genetic engineering[J].Guizhou Agricultural Science,2014,(12):37-42.
[37] THOMASHOW M F.Role of cold-responsive genes in plant freezing tolerance[J].Plant Physiology,1998,118(1):1-7.