花生叶片旱后恢复生长的生理特征及相关基因表达的研究
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摘要
为研究花生叶片在干旱胁迫后恢复生长的生理和相关基因表达,本论文采用不同浓度聚乙二醇6000(PEG)模拟干旱条件处理花生幼苗,复水后观察花生幼苗恢复生长的能力,并分析在干旱和复水过程中叶片含水量、叶绿素含量、ABA含量以及相关基因表达的变化。结果表明, 10%PEG处理花生幼苗24 h后复水,不同叶片旱后恢复的能力不同,嫩叶较老叶恢复能力强;部分展开叶片在干旱和复水后叶绿素含量均高于正常水平;而全展开叶片中叶绿素含量在干旱时升高,复水后降低;干旱时,幼叶中的ABA含量显著上升,复水后降低。而老叶(成熟叶)中的ABA含量持续降低; AhATL1、AhUGT、AhCYP707A1、AhGLK1和AhPORA的表达在干旱时降低,复水后上调。AhNCED1和AhBG的表达在干旱时升高,复水后降低。在不同叶片中, 7个基因的变化规律不完全一致,表明不同叶片响应干旱和复水恢复生长的能力可能与这些基因的表达差异有关。
To study the physiological and molecular basis of Arachis hypogaea(peanut) leaves during recovery after drought, different concentrations of polyethylene glycol 6000(PEG) were used to simulate drought condition. Peanut seedlings were treated by PEG, and the recovery ability of peanut seedlings was observed. The relative water content, chlorophyll content, ABA content and the expression of seven genes were analyzed after drought and rewatering. The results showed that 10% PEG was appropriate to treat peanut seedlings for 24 h. The recovery ability of different leaves during recovery from drought was different, showing that young leaves were better than old leaves. The chlorophyll contents of gently unfolded leaves in drought and rewatering process were higher than that of normal condition. However, the chlorophyll content of completely unfolded was increased during drought but decreased after rewatering. ABA content of young leaves was increased significantly during drought, but reduced after rewatering, while the content of ABA in old leaves(mature leaves) was decreased continuously. The expressions of AhATL1, AhUGT, AhCYP707 A1, AhGLK1 and AhPORA were decreased during drought and up-regulated after rewatering. In contrast, the expression of AhNCED1 and AhBG were increased during drought but decreased after rewartering. The expression trend of seven genes was inconsistent in different leaves, indicating that the recovery ability of different leaves may be related to the different expressions of these genes.
To study the physiological and molecular basis of Arachis hypogaea(peanut) leaves during recovery after drought, different concentrations of polyethylene glycol 6000(PEG) were used to simulate drought condition. Peanut seedlings were treated by PEG, and the recovery ability of peanut seedlings was observed. The relative water content, chlorophyll content, ABA content and the expression of seven genes were analyzed after drought and rewatering. The results showed that 10% PEG was appropriate to treat peanut seedlings for 24 h. The recovery ability of different leaves during recovery from drought was different, showing that young leaves were better than old leaves. The chlorophyll contents of gently unfolded leaves in drought and rewatering process were higher than that of normal condition. However, the chlorophyll content of completely unfolded was increased during drought but decreased after rewatering. ABA content of young leaves was increased significantly during drought, but reduced after rewatering, while the content of ABA in old leaves(mature leaves) was decreased continuously. The expressions of AhATL1, AhUGT, AhCYP707 A1, AhGLK1 and AhPORA were decreased during drought and up-regulated after rewatering. In contrast, the expression of AhNCED1 and AhBG were increased during drought but decreased after rewartering. The expression trend of seven genes was inconsistent in different leaves, indicating that the recovery ability of different leaves may be related to the different expressions of these genes.
引文
Arnon DI(1949).Copper enzymes in isolated chloroplasts.polyphenol oxidase in Beta vulgaris.Plant Physiol,24:1-15
Chaves MM,Flexas J,Pinheiro C(2009).Photosynthesis under drought and salt stress:regulation mechanisms from whole plant to cell.Ann Bot,103:551-560
Duan M,Zhang RX,Zhu FG,et al(2017).A lipid-anchored NAC transcription factor is translocated into the nucleus and activates glyoxalase I expression during drought stress.Plant Cell,29:1748-1772
Duan Na,Wang Jia,Liu Fang,et al(2018).Research progress on drought resistance of plant.Mol Plant Breed,16:5093-5099(in Chinese with English abstract)[段娜,王佳,刘芳等(2018).植物抗旱性研究进展.分子植物育种,16:5093-5099]
Fang Y,Xiong L(2015).General mechanisms of drought response and their application in drought resistance improvement in plants.Cell Mol Life Sci,72:673-689
Fukao T,Yeung E,Baileyserres J(2011).The submergence tolerance regulator SUB1A mediates crosstalk between submergence and drought tolerance in rice.Plant Cell,23:412-427
Ge K,Liu X,Li X,et al(2017).Isolation of an ABA Transporter-Like 1 gene from Arachis hypogaea that affects ABA import and reduces ABA sensitivity in Arabidopsis.Front Plant Sci,8:1150
Hayashi M,Kinoshita T(2011).Crosstalk between blue-lightand aba-signaling pathways in stomatal guard cells.Plant Signal Behav,6:1662-1664
Li XY,Liu X,Yao Y,et al(2013).Overexpression of Arachis hypogaea AREB1 gene enhances drought tolerance by modulating ROS scavenging and maintaining endogenous ABA content.Int J Mol Sci,14:12827-12842
Liu JL,Zhao CX,Wu N,et al(2011).Effects of drought and rewatering at seedling stage on photosynthetic characteristics and water use efficiency of peanut.Sci Agric Sin,44:469-476(in Chinese with English abstract)[刘吉利,赵长星,吴娜等(2011).苗期干旱及复水对花生光合特性及水分利用效率的影响.中国农业科学,44:469-476]
Liu JS,Li L(2006).Drought resistance of different peanut and its relationship with endogenous ABA.Plant Physiol Commun,42:1115-1116[刘吉升,李玲(2006).不同品种花生的抗旱能力及其与内源ABA的关系.植物生理通讯,42:469-476]
Liu X,Hong L,Li,XY,et al(2011).Improved drought and salt tolerance in transgenic Arabidopsis overexpressing a NAC transcriptional factor from Arachis hypogaea.Biosci Biotech Bioch,75:443-450
Liu X,Li LM,Li MJ,et al(2018).AhGLK1 affects chlorophyll biosynthesis and photosynthesis in peanut leaves during recovery from drought.Sci Rep,8:2250
Liu X,Liu S,Wu J,et al(2013).Overexpression of Arachis hypogaea NAC3 in tobacco enhances dehydration and drought tolerance by increasing superoxide scavenging.Plant Physiol Bioc,70:354-359
Morimoto K,Ohama N,Kidokoro S,et al(2017).BPM-CUL3E3 ligase modulates thermo tolerance by facilitating negative regulatory domain-mediated degradation of DREB2A in Arabidopsis.Proc Natl Acad Sci USA,114:8528-8536
Murmu J,Wilton M,Allard G,et al(2014).Arabidopsis GOLDEN2-LIKE(GLK)transcription factors activate jasmonic acid(JA)-dependent disease susceptibility to the biotrophic pathogen Hyaloperonospora arabidopsidis,as well as JA-independent plant immunity against the necrotrophic pathogen Botrytis cinerea.Mol Plant Pathol,15:174-184
Ni FR,Wu L,Wang Q,et al(2017).Turnip yellow mosaic virus P69 interacts with and suppresses GLK transcription factors to cause pale-green symptoms in Arabidopsis.Mol Plant,10:764-766
Rauf M,Arif M,Dortay H.et al(2013).ORE1 balances leaf senescence against maintenance by antagonizing G2-like-mediated transcription.EMBO Rep,14:382-388
Saito S,Hirai N,Matsumoto C,et al(2004).Arabidopsis CY-P707As encode(+)-abscisic acid 8′-hydroxylase,a key enzyme in the oxidative catabolism of abscisic acid.Plant Physiol,134:1439-1449
Toh S,Imamura A,Watanabe A,et al(2008).High temperature-induced ABA biosynthesis and its role in the inhibition of GAS action in Arabidopsis seeds.Plant Physiol,146:1368-1385
Tamburino R,Vitale M,Ruggiero A,et al(2017).Chloroplast proteome response to drought stress and recovery in tomato(Solanum lycopersicum L.).BMC Plant Biol,17:40
VankováR,DobráJ,?torchováH(2012).Recovery from drought stress in tobacco.Plant Signal Behav,7:19-21
Verslues PE,Agarwal M,Katiyar-Agarwal S,et al(2016).Methods and concepts in quantifying resistance to drought,salt and freezing,abiotic stresses that affect plant water status.Plant J,45:523-539
Wan X,Li L(2005).Molecular cloning and characterization of a dehydration-inducible cDNA encoding a putative9-cis-epoxycarotenoid dioxygenase in Arachis hygogaea L.Dna Sequence,16:217-223
Waters MT,Wang P,Korkaric M,et al(2009).GLK transcription factors coordinate expression of the photosynthetic apparatus in Arabidopsis.Plant Cell,21:1109-1128
Xu Z,Zhou G,Shimizu H(2010).Plant responses to drought and rewatering.Plant Signal Behav,5:649-654
Chaves MM,Flexas J,Pinheiro C(2009).Photosynthesis under drought and salt stress:regulation mechanisms from whole plant to cell.Ann Bot,103:551-560
Duan M,Zhang RX,Zhu FG,et al(2017).A lipid-anchored NAC transcription factor is translocated into the nucleus and activates glyoxalase I expression during drought stress.Plant Cell,29:1748-1772
Duan Na,Wang Jia,Liu Fang,et al(2018).Research progress on drought resistance of plant.Mol Plant Breed,16:5093-5099(in Chinese with English abstract)[段娜,王佳,刘芳等(2018).植物抗旱性研究进展.分子植物育种,16:5093-5099]
Fang Y,Xiong L(2015).General mechanisms of drought response and their application in drought resistance improvement in plants.Cell Mol Life Sci,72:673-689
Fukao T,Yeung E,Baileyserres J(2011).The submergence tolerance regulator SUB1A mediates crosstalk between submergence and drought tolerance in rice.Plant Cell,23:412-427
Ge K,Liu X,Li X,et al(2017).Isolation of an ABA Transporter-Like 1 gene from Arachis hypogaea that affects ABA import and reduces ABA sensitivity in Arabidopsis.Front Plant Sci,8:1150
Hayashi M,Kinoshita T(2011).Crosstalk between blue-lightand aba-signaling pathways in stomatal guard cells.Plant Signal Behav,6:1662-1664
Li XY,Liu X,Yao Y,et al(2013).Overexpression of Arachis hypogaea AREB1 gene enhances drought tolerance by modulating ROS scavenging and maintaining endogenous ABA content.Int J Mol Sci,14:12827-12842
Liu JL,Zhao CX,Wu N,et al(2011).Effects of drought and rewatering at seedling stage on photosynthetic characteristics and water use efficiency of peanut.Sci Agric Sin,44:469-476(in Chinese with English abstract)[刘吉利,赵长星,吴娜等(2011).苗期干旱及复水对花生光合特性及水分利用效率的影响.中国农业科学,44:469-476]
Liu JS,Li L(2006).Drought resistance of different peanut and its relationship with endogenous ABA.Plant Physiol Commun,42:1115-1116[刘吉升,李玲(2006).不同品种花生的抗旱能力及其与内源ABA的关系.植物生理通讯,42:469-476]
Liu X,Hong L,Li,XY,et al(2011).Improved drought and salt tolerance in transgenic Arabidopsis overexpressing a NAC transcriptional factor from Arachis hypogaea.Biosci Biotech Bioch,75:443-450
Liu X,Li LM,Li MJ,et al(2018).AhGLK1 affects chlorophyll biosynthesis and photosynthesis in peanut leaves during recovery from drought.Sci Rep,8:2250
Liu X,Liu S,Wu J,et al(2013).Overexpression of Arachis hypogaea NAC3 in tobacco enhances dehydration and drought tolerance by increasing superoxide scavenging.Plant Physiol Bioc,70:354-359
Morimoto K,Ohama N,Kidokoro S,et al(2017).BPM-CUL3E3 ligase modulates thermo tolerance by facilitating negative regulatory domain-mediated degradation of DREB2A in Arabidopsis.Proc Natl Acad Sci USA,114:8528-8536
Murmu J,Wilton M,Allard G,et al(2014).Arabidopsis GOLDEN2-LIKE(GLK)transcription factors activate jasmonic acid(JA)-dependent disease susceptibility to the biotrophic pathogen Hyaloperonospora arabidopsidis,as well as JA-independent plant immunity against the necrotrophic pathogen Botrytis cinerea.Mol Plant Pathol,15:174-184
Ni FR,Wu L,Wang Q,et al(2017).Turnip yellow mosaic virus P69 interacts with and suppresses GLK transcription factors to cause pale-green symptoms in Arabidopsis.Mol Plant,10:764-766
Rauf M,Arif M,Dortay H.et al(2013).ORE1 balances leaf senescence against maintenance by antagonizing G2-like-mediated transcription.EMBO Rep,14:382-388
Saito S,Hirai N,Matsumoto C,et al(2004).Arabidopsis CY-P707As encode(+)-abscisic acid 8′-hydroxylase,a key enzyme in the oxidative catabolism of abscisic acid.Plant Physiol,134:1439-1449
Toh S,Imamura A,Watanabe A,et al(2008).High temperature-induced ABA biosynthesis and its role in the inhibition of GAS action in Arabidopsis seeds.Plant Physiol,146:1368-1385
Tamburino R,Vitale M,Ruggiero A,et al(2017).Chloroplast proteome response to drought stress and recovery in tomato(Solanum lycopersicum L.).BMC Plant Biol,17:40
VankováR,DobráJ,?torchováH(2012).Recovery from drought stress in tobacco.Plant Signal Behav,7:19-21
Verslues PE,Agarwal M,Katiyar-Agarwal S,et al(2016).Methods and concepts in quantifying resistance to drought,salt and freezing,abiotic stresses that affect plant water status.Plant J,45:523-539
Wan X,Li L(2005).Molecular cloning and characterization of a dehydration-inducible cDNA encoding a putative9-cis-epoxycarotenoid dioxygenase in Arachis hygogaea L.Dna Sequence,16:217-223
Waters MT,Wang P,Korkaric M,et al(2009).GLK transcription factors coordinate expression of the photosynthetic apparatus in Arabidopsis.Plant Cell,21:1109-1128
Xu Z,Zhou G,Shimizu H(2010).Plant responses to drought and rewatering.Plant Signal Behav,5:649-654