东北草地野大麦对混合盐碱胁迫的生理响应及转录组分析
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摘要
为了研究盐碱胁迫下野大麦(Hordeum brevisubulatum)的生理响应及耐盐碱基因的发掘,采用不同浓度NaCl与NaHCO3混合液胁迫处理45日龄幼苗,通过理化分析方法确定野大麦盐碱逆境胁迫临界值,通过转录组测序技术(RNA-Seq)分析基因表达情况。结果表明,不同浓度(100、200、300、400、500 mmol·L~(–1))混合盐碱胁迫2 d后,野大麦叶片可溶性蛋白含量、超氧化物歧化酶(SOD)活性呈先升高后降低的趋势,均在300 mmol·L~(–1)达到最大值,脯氨酸、可溶性糖含量呈连续上升趋势。选取300 mmol·L~(–1)处理组的野大麦叶片进行转录组测序分析,与对照组(无盐碱处理)相比有4 163个基因上调,1 936个基因下调;对差异表达基因进行GO功能分类,可分为生物过程、细胞组分和分子功能3个主类52个小类;4 405个差异基因被注释到132个通路中,主要涉及蔗糖合成、脯氨酸合成、过氧化物酶体等代谢途径,蔗糖合成酶(SuS)、鸟氨酸转氨酶(OAT)、谷氨酸激酶(ProB)、谷氨酸-5-半醛脱氢酶(ProA)、超氧化物歧化酶(SOD)等的编码基因上调表达。野大麦可以通过提高某些渗透调节物质含量适应盐碱逆境胁迫,这种代谢过程调节蕴涵相关基因表达调控的生理机制。
In order to study the physiological response and salt tolerance genes of wild barley(Hordeum brevisubulatum)under saline alkali stress, 45-day-old seedlings were treated with different concentrations of NaCl and NaHCO_3. The critical value of saline alkali stress was determined by physicochemical analysis. The gene expression was analysed by RNA-Seq.The soluble protein content and superoxide dismutase(SOD) activity of wild barley leaves increased first and then decreased after treatment with different concentrations of NaCl and NaHCO3(100, 200, 300, 400, 500 mmol·L~(–1)) for 2 days. The maximum value was reached at 300 mmol·L~(–1), and the content of proline and soluble sugar showed a continuous upward trend. The wild barley leaves in the 300 mmol·L~(–1) treatment group were sequenced and analysed. Compared with the control group(treatment of salt and alkali free), 4 163 genes were up-regulated and 1 936 genes were down-regulated. The Gene Ontology(GO) function classification of the differentially expressed genes could be divided into 52 subclasses of three main categories: biological process, cell component, and molecular function. A total of 4 405 differential genes were annotated into 132 pathways, mainly involved in sucrose synthesis, proline synthesis, peroxisome metabolic pathways. The genes encoding sucrose synthase(SuS), ornithine aminotransferase(OAT), glutamate kinase(proB), glutamate-5-semialdehyde dehydrogenase(proA), superoxide dismutase(SOD) were up-regulated. Wild barley can adapt to saline alkali stress by increasing the content of some osmotic regulators, this metabolic process regulates the physiological mechanism of regulating the expression of related genes.
In order to study the physiological response and salt tolerance genes of wild barley(Hordeum brevisubulatum)under saline alkali stress, 45-day-old seedlings were treated with different concentrations of NaCl and NaHCO_3. The critical value of saline alkali stress was determined by physicochemical analysis. The gene expression was analysed by RNA-Seq.The soluble protein content and superoxide dismutase(SOD) activity of wild barley leaves increased first and then decreased after treatment with different concentrations of NaCl and NaHCO3(100, 200, 300, 400, 500 mmol·L~(–1)) for 2 days. The maximum value was reached at 300 mmol·L~(–1), and the content of proline and soluble sugar showed a continuous upward trend. The wild barley leaves in the 300 mmol·L~(–1) treatment group were sequenced and analysed. Compared with the control group(treatment of salt and alkali free), 4 163 genes were up-regulated and 1 936 genes were down-regulated. The Gene Ontology(GO) function classification of the differentially expressed genes could be divided into 52 subclasses of three main categories: biological process, cell component, and molecular function. A total of 4 405 differential genes were annotated into 132 pathways, mainly involved in sucrose synthesis, proline synthesis, peroxisome metabolic pathways. The genes encoding sucrose synthase(SuS), ornithine aminotransferase(OAT), glutamate kinase(proB), glutamate-5-semialdehyde dehydrogenase(proA), superoxide dismutase(SOD) were up-regulated. Wild barley can adapt to saline alkali stress by increasing the content of some osmotic regulators, this metabolic process regulates the physiological mechanism of regulating the expression of related genes.
引文
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[15]成铁龙,李焕勇,武海雯,刘正祥,武香,杨升,张华新,杨秀艳.盐胁迫下4种耐盐植物渗透调节物质积累的比较.林业科学研究,2015,28(6):826-832.CHENG T L, LI H Y, WU H W, LIU Z X, WU X, YANG S, ZHANG X H, YANG X Y. Comparison on osmotica accumulation of different salt-tolerant plants under salt stress. Forest Research, 2015, 28(6):826-832.
[16]刘凤歧,刘杰淋,朱瑞芬,张悦,郭勇,韩贵清,唐凤兰.4种燕麦对NaCl胁迫的生理响应及耐盐性评价.草业学报,2015, 24(1):183-189.LIU F Q, LIU J, ZHU R F, ZHANG Y, GUO Y, HAN G Q, TANG F L. Physiological responses and tolerance of four oat varieties to salt stress. Acta Prataculturae Sinica, 2015, 24(1):183-189.
[17]赵江涛,李晓峰,李航,徐睿态.可溶性糖在高等植物代谢调节中的生理作用.安徽农业科学,2006(24):6423-6425.ZHAO J T, LI X F, LI H, XU R M. Research on the role of the soluble sugar in the regulation of physiological metabolism in higher plant. Journal of Anhui Agricultural Sciences, 2006(24):6423-6425.
[18] PRADO F E,BOERO C,GALLARDO M, GONZALEZ J A. Effect of NaCl on germination,growth,and soluble sugar content in Chenopodium quinoa willd.seeds. Botanical Bulletin of Academia Sinica, 2000, 41(1):27-34.
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