野大麦对干旱胁迫的生理响应与转录组分析
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摘要
为了研究野大麦在干旱胁迫下的生理响应以及发掘耐旱性相关基因,对不同浓度聚乙二醇(PEG) 6000胁迫条件下的野大麦幼苗进行生理指标测定及转录组测序分析。结果表明:不同浓度PEG 6000胁迫2 d后,野大麦叶片脯氨酸含量呈连续上升趋势,可溶性糖及超氧化物歧化酶(SOD)活性呈先上升后下降趋势。通过隶属函数法选取30 mmol·L~(-1)PEG 6000处理组野大麦叶片进行转录组分析测序,与对照组相比上调表达基因6 868个,下调表达基因2 081个;对差异表达基因进行GO(gene ontology,基因本体)功能分类,可以分为3大类并包含54个功能组;通过KEGG通路富集将6 579个差异表达基因富集到136个通路中,主要包括精氨酸和脯氨酸代谢、糖酵解和糖异生、淀粉和蔗糖代谢、过氧化物酶体等途径,并发掘了各个代谢途径中相关上调表达基因。
The physiological response of Hordeum brevisubulatum to drought stress was researched so as to identify the drought tolerance related to the genes,physiological indexes and transcriptome sequencing of H. brevisubulatum seedlings under different concentrations of polyethylene glycol (PEG) 6000 stress. The results showed that the content of proline in leaves of H. brevisubulatum was increased continuously after the PEG 6000 stress for 2 days,and the activity of soluble sugar and superoxide dismutase (SOD) was increased at first and then decreased. The membership function method was used to select the transcriptome analysis and sequencing of H. brevisubulatum leaves under the treatment of 30 mmol·L~(-1) PEG 6000. Compared with the control group,the 6 868 up-regulated genes and 2 081 down-regulated genes were identified. The differentially expressed genes were classified into three categories and 54 functional groups by Go function classification. The 6 579 differentially expressed genes were enriched into 136 pathways by KEGG pathway enrichment. Arginine and proline metabolism,glycolysis and gluconeogenesis,starch and sucrose metabolism,peroxisome pathways and other pathways,and the related up-regulated expression genes in various metabolic pathways were discovered.
The physiological response of Hordeum brevisubulatum to drought stress was researched so as to identify the drought tolerance related to the genes,physiological indexes and transcriptome sequencing of H. brevisubulatum seedlings under different concentrations of polyethylene glycol (PEG) 6000 stress. The results showed that the content of proline in leaves of H. brevisubulatum was increased continuously after the PEG 6000 stress for 2 days,and the activity of soluble sugar and superoxide dismutase (SOD) was increased at first and then decreased. The membership function method was used to select the transcriptome analysis and sequencing of H. brevisubulatum leaves under the treatment of 30 mmol·L~(-1) PEG 6000. Compared with the control group,the 6 868 up-regulated genes and 2 081 down-regulated genes were identified. The differentially expressed genes were classified into three categories and 54 functional groups by Go function classification. The 6 579 differentially expressed genes were enriched into 136 pathways by KEGG pathway enrichment. Arginine and proline metabolism,glycolysis and gluconeogenesis,starch and sucrose metabolism,peroxisome pathways and other pathways,and the related up-regulated expression genes in various metabolic pathways were discovered.
引文
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[19]Scandalios J G.Oxygen stress and superoxide dismutase[J].plant physiology,1993,101(1):7-11.
[20]李雪,田新会,杜文华.小黑麦品系成株期抗旱性研究[J].核农学报,2018,32(2):377-388.[Li Xue,Tian Xinhui,Du Wenhua.Study on drought resistance of triticale lines at the adult plant stage[J].Journal of Nuclear Agricultural Sciences,2018,32(2):377-388.]
[21]Kishor P B K,Sangam S,Amrutha R N,et al.Regulation of proline biosynthesis,degradation,uptake and transport in higher plants:Its implications in plant growth and abiotic stress tolerance[J].Current Science,2005,54(4):271-275.
[22]刘家林,欧阳林娟,曾嘉丽,等.水稻SOD基因家族的全基因组分析及逆境胁迫下表达研究[J].分子植物育种,2018,16(9):2 753-2 760.[Liu Jialin,Ouyang Linjuan,Zeng Jiali,et al.Genome-wide analysis of rice SOD gene family and expression research under stress[J].Molecular Plant Breeding,2018,16(9):2 753-2 760.]
[2]刘晓静,马东来,张继权,等.辽西北地区玉米干旱的识别方法[J].干旱区研究,2018,35(3):722-727.[Liu Xiaojing,Ma Donglai,Zhang Jiquan,et al.Identification method of maize drought in Northwest Liaoning Province[J].Arid Zone Research,2018,35(3):722-727.]
[3]Fang Y J,Xiong L Z.General mechanisms of drought response and their application in drought resistance improvement in plants[J].Cellular and Molecular Life Sciences,2015,72(4):673-689.
[4]张文超,王玉凤,张翼飞,等.耕作方式对松嫩平原半干旱区土壤养分含量和玉米产量的影响[J].作物杂志,2017(4):123-128.[Zhang Wenchao,Wang Yufeng,Zhang Yifei,et al.Effects of different tillage methods on changes of soil nutrients and grain yield of maize in semi-arid regions of Songnen Plain[J].Crops,2017(4):123-128.]
[5]张玉红,张树清,苏立英.基于3S的扎龙湿地土地盐碱化趋势分析.农业系统科学与综合研究,2010,26(2):140-144.[Zhang Yuhong,Zhang Shuqing,Su Liying.Analysis of basification trend at Zhalong Wetland based on 3S[J].System Sciences and Comprehensive Studies in Agriculture,2010,26(2):140-144.]
[6]罗金明,王永洁,柏林,等.扎龙盐沼湿地旱化特征及其对演替的影响[J].干旱区研究,2018,35(5):1 167-1 172.[Luo Jinming,Wang Yongjie,Bai Lin,et al.Effect of drought stress on succession of the Zhalong Wetland[J].Arid Zone Research,2018,35(5):1 167-1 172.]
[7]焦德志,宋士伟.扎龙湿地野大麦对Na Cl胁迫的生理响应[J].黑龙江畜牧兽医,2018(15):147-150.[Jiao Dezhi,Song Shiwei.The physiological response of Hordeum brevisubulatum to Na Cl stress in Zhalong Wetland[J].Heilongjiang Animal Science and Veterinary Medicine,2018(15):147-150.]
[8]张小芳,王冰冰,徐燕,等.PEG模拟干旱胁迫下野生大豆转录组分析[J].大豆科学,2018(5):681-689.[Zhang Xiaofang,Wang Bingbing,Xu Yan,et al.The Transcriptome analysis of wild soybean under drought stress simulated by PEG[J].Soybean Science,2018(5):681-689.]
[9]冷暖,刘晓巍,张娜,等.草地早熟禾干旱胁迫转录组差异性分析[J].草业学报,2017,26(12):128-137.[Leng Nuan,Liu Xiaowei,Zhang Na,et al.Differential gene analysis of Poa pratensis in response to drought stress[J].Acta Prataculturae Sinica,2017,26(12):128-137.]
[10]王丹,龚荣高.干旱胁迫下枇杷叶片的转录组分析[J].华北农学报,2017,32(1):60-67.[Wang Dan,Gong Ronggao.Transcriptome profiling of Loquat Leaves under drought stress[J].Acta Agriculturae Boreali-Sinica,2017,32(1):60-67.]
[11]刘志洋.游离脯氨酸含量测定法比较宿根花卉抗寒性[J].黑龙江农业科学,2016(11):102-105.[Liu Zhiyang.Cold resistance comparison of perennial flowers by determination of free proline content[J].Heilongjiang Agricultural Sciences,2016(11):102-105.]
[12]王法琴,陆兔林,毛春芹,等.3种比色法测定五味子中多糖[J].中成药,2015,37(4):814-818.[Wang Faqin,Lu Tulin,Mao Chunqin,et al.Determination of polysaccharides from Schisandrae chinensis Fructus by three colorimetric method[J].Chinese Traditional Patent Medicine,2015,37(4):814-818.]
[13]曲敏,秦丽楠,刘羽佳,等.两种检测SOD酶活性方法的比较[J].食品安全质量检测学报,2014,5(10):3 318-3 323.[Qu Min,Qin Linan,Liu Yujia,et al.The comparison of two methods of testing superoxide dismutase activity[J].Journal of Food Safety&Quality,2014,5(10):3 318-3 323.]
[14]赵江涛,李晓峰,李航,等.可溶性糖在高等植物代谢调节中的生理作用[J].安徽农业科学,2006,34(24):6 423-6 425,6 427.[Zhao Jiangtao,Li Xiaofeng,Li Hang,et al.Research on the role of the soluble sugar in the regulation of physiological metabolism in higher plant[J].Journal of Anhui Agricultural Sciences,2006,34(24):6 423-6 425,6427.]
[15]冯淑华,陈雅君,周阳.干旱胁迫下三叶草渗透调节物质动态及与叶片水分的相关性[J].中国草地学报,2011,33(6):70-74.[Feng Shuhua,Chen Yajun,Zhou Yang.Correlation analysis between dynamic change of osmoregulation substance and moisture in leaves of clovers under drought stress[J].Chinese Journal of Grassland,2011,33(6):70-74.]
[16]刘艳,陈贵林,蔡贵芳,等.干旱胁迫对甘草幼苗生长和渗透调节物质含量的影响[J].西北植物学报,2011,31(11):2 259-2 264.[Liu Yan,Chen Guilin,Cai Guifang,et al.Growth and osmoregulation substances accumulation of Glycyrrhiza uralensis seedling under drought stress[J].Acta Botanica Boreali-Occidentalia Sinica,2011,31(11):2 259-2 264.]
[17]许翩翩,王建柱.三种常见边坡植物对模拟干旱环境抗旱性能的研究[J].草业学报,2018,27(2):36-47.[Xu Pianpian,Wang Jianzhu.Drought resistance of three common slope plants determined in a simulated drought experiment[J].Acta Prataculturae Sinica,2018,27(2):36-47]
[18]崔慧萍,周薇,郭长虹.植物过氧化物酶体在活性氧信号网络中的作用[J].中国生物化学与分子生物学报,2017,33(3):220-226.[Cui Huiping,Zhou Wei,Guo Changhong.The role of plant peroxisomes in ROS signalling network[J].Chinese Journal of Biochemistry and Molecular Biology,2017,33(3):220-226.]
[19]Scandalios J G.Oxygen stress and superoxide dismutase[J].plant physiology,1993,101(1):7-11.
[20]李雪,田新会,杜文华.小黑麦品系成株期抗旱性研究[J].核农学报,2018,32(2):377-388.[Li Xue,Tian Xinhui,Du Wenhua.Study on drought resistance of triticale lines at the adult plant stage[J].Journal of Nuclear Agricultural Sciences,2018,32(2):377-388.]
[21]Kishor P B K,Sangam S,Amrutha R N,et al.Regulation of proline biosynthesis,degradation,uptake and transport in higher plants:Its implications in plant growth and abiotic stress tolerance[J].Current Science,2005,54(4):271-275.
[22]刘家林,欧阳林娟,曾嘉丽,等.水稻SOD基因家族的全基因组分析及逆境胁迫下表达研究[J].分子植物育种,2018,16(9):2 753-2 760.[Liu Jialin,Ouyang Linjuan,Zeng Jiali,et al.Genome-wide analysis of rice SOD gene family and expression research under stress[J].Molecular Plant Breeding,2018,16(9):2 753-2 760.]