中国农业科学 ›› 2019, Vol. 52 ›› Issue (22): 4002-4015.doi: 10.3864/j.issn.0578-1752.2019.22.006

• 分子遗传 • 上一篇    下一篇

不同耐盐性高粱在盐逆境下的比较转录组分析

张飞,王艳秋,朱凯,张志鹏,朱振兴,卢峰(),邹剑秋()   

  1. 辽宁省农业科学院高粱研究所,沈阳110161
  • 收稿日期:2019-06-14 接受日期:2019-08-12 出版日期:2019-11-16 发布日期:2019-11-16
  • 通讯作者: 卢峰,邹剑秋
  • 作者简介:张飞,Tel:024-31029903;E-mail:zhangfei19821121@163.com
  • 基金资助:
    国家现代农业产业技术体系(CARS-06-13.5-A11);国家现代农业产业技术体系(CARS-06-13.5-A22);辽宁省自然科学基金(2019-MS-197)

Comparative Transcriptome Analysis of Different Salt Tolerance Sorghum (Sorghum bicolor L. Moench) Under Salt Stress

ZHANG Fei,WANG YanQiu,ZHU Kai,ZHANG ZhiPeng,ZHU ZhenXing,LU Feng(),ZOU JianQiu()   

  1. Sorghum Institute, Liaoning Academy of Agricultural Sciences, Shenyang 110161
  • Received:2019-06-14 Accepted:2019-08-12 Online:2019-11-16 Published:2019-11-16
  • Contact: Feng LU,JianQiu ZOU

摘要:

【目的】土壤盐渍化是制约作物生产的重要非生物胁迫因子之一,高粱耐盐性强,进行高粱耐盐基因挖掘及分子机制研究是开发和利用盐渍土壤的有效途径,通过转录组测序分析与高粱耐盐相关的基因调控机制和代谢通路,挖掘高粱耐盐潜力。【方法】 通过以筛选出的极耐盐品种八叶齐和盐极敏感品种PL212为试验材料,采用盆栽沙培,在播种后20 d(5叶期)采用180 mmol·L -1的 NaCl 溶液漫灌模拟盐逆境,盐胁迫48 h后取幼叶,并连同对照(未经过盐处理)的同期幼苗共4个样品提取RNA,进行转录组测序,采用qRT-PCR方法对测序结果进行验证。 【结果】 耐盐和盐敏感材料分别在盐渍和非盐渍处理下的4个样品间共检测到1 338个差异表达基因,包括819个上调基因和519个下调基因。聚类分析发现在应答盐渍胁迫逆境时,5个依赖性氧合酶超家族蛋白、4个富含半胱氨酸的激酶、3个谷胱甘肽S-转移酶和3个重金属运输/解毒超家族蛋白相关基因表现出明显的上调表达和下调表达,还发现1个K +转运蛋白基因在耐盐调节中起着重要作用。GO分析发现在15 418个基因中获得4 528个有效GO注释条目,同时耐盐和盐敏感材料在遭受盐逆境时的生物过程、细胞组分和分子功能3个方面均存在较大差异。生物过程中代谢过程、细胞过程耐盐材料明显高于盐敏感材料,耐盐材料的生理过程中较盐敏感材料增加了多生物过程和定位这两个过程,很可能与耐盐材料盐抗性较强密切相关。差异基因KEGG分析结果显示耐盐和盐敏感材料在对照和盐渍胁迫条件下的苯丙烷类生物合成、苯丙氨酸代谢、类黄酮生物合成3个途径中差异基因表达较多,可能是造成耐盐和盐敏感材料耐盐性差异较大的重要原因。 【结论】 高粱耐盐调控基因表达涉及生物过程、细胞组分和分子功能多个方面,生物过程和定位这两个过程是提高高粱耐盐性的关键;苯丙烷类生物合成、苯丙氨酸代谢、类黄酮生物合成3个途径的基因表达很可能是造成盐害的重要原因。

关键词: 高粱, 耐盐, 转录组, 差异基因表达, 生理调控

Abstract:

【Objective】Soil salinization is one of the important abiotic stress factors that restricts crop production. Understanding the salt-tolerant mechanism of sorghum may provide a novel avenue to utilize saline soil for sorghum production. The objective of this study was to explore gene regulation mechanisms and metabolic pathways that related to salt tolerance of sorghum by transcriptome sequencing. 【Method】 The salt-tolerant genotype Bayeqi and salt-sensitive genotype PL212 were planted in plastic pots. At five-leaf stage (20 days after sowing), plants were treated with 180 mmol L -1 NaCl. Forty-eight hours after treatment, leaves treated by NaCl and unstressed control were sampled and were used for RNA extraction and transcriptome sequencing. Sequencing results were verified by qRT-PCR. 【Result】 Results showed that a total of 1 338 deferentially expressed genes, including 819 up-regulated and 519 down-regulated genes were detected. Cluster analysis revealed that in response to salt stress, five dependent oxygenase superfamily proteins, four cysteine-rich RLKs, three Glutathione S-transferase and three heavy metal transport/detoxification superfamily protein-related genes were significant up-regulated and/or down-regulated, and one K + ion transporter gene was also found to play an important role in salt-tolerance regulation. GO analysis found that 4 528 valid GO annotation entries were obtained from 15 418 genes, and salt-tolerant and salt-sensitive materials showed significant difference in biological processes, cellular components and molecular functions under salt stress treatment. The salt-tolerant materials exhibited obviously higher metabolic processes and cellular processes than salt-sensitive materials. Compared with salt-sensitive materials, multiple biological processes and localization processes were increased in salt-tolerant genotype, which might be the reasons of salt-tolerance. KEGG analysis showed that the salt-tolerant and salt-sensitive materials had more differential gene expression in phenylpropanoid biosynthesis, phenylalanine metabolism and flavonoid biosynthesis under control and salt stress conditions, which may be an important reason for the weak salt tolerance of sensitive materials. 【Conclusion】 The expression of salt-tolerant genes in sorghum is involved in many aspects of biological processes, cellular components and molecular functions. The gene expression in multiple processes and localization processes contributes to the salt tolerance, while excessive gene expression in phenylpropanoid biosynthesis, phenylalanine metabolism, and flavonoid biosynthesis likely contributes to the damage under salt stress.

Key words: sorghum, salt tolerance, transcriptome, differential gene expression, physiological regulation