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深度测序抑制性消减文库鉴定出受盐胁迫影响的甜高粱差异表达转录本。

Deep sequencing of suppression subtractive library identifies differentially expressed transcripts of Saccharum spontaneum exposed to salinity stress.

机构信息

Division of Crop Improvement, ICAR-Sugarcane Breeding Institute, Coimbatore, Tamil Nadu, India.

Biosystematics Group, Wageningen University & Research, Wageningen, The Netherlands.

出版信息

Physiol Plant. 2022 Jan;174(1):e13645. doi: 10.1111/ppl.13645.

DOI:10.1111/ppl.13645
PMID:35112353
Abstract

Saccharum spontaneum, a wild relative of sugarcane, is highly tolerant to drought and salinity. The exploitation of germplasm resources for salinity tolerance is a major thrust area in India. In this study, we utilized suppression subtractive hybridization (SSH) followed by sequencing for the identification of upregulated transcripts during salinity stress in S. spontaneum clones coming from different geographical regions of India. Our sequencing of the SSH library revealed that 95% of the transformants contained inserts of size 200-1500 bp. We have identified 314 differentially expressed transcripts in the salinity-treated samples after subtraction, which were subsequently validated by quantitative real-time polymerase chain reaction. Functional annotation and pathway analysis revealed that the upregulated transcripts were a result of protein modifications, stress, and hormone signaling along with cell wall development and lignification. The prominently upregulated transcripts included UDP glucose dehydrogenase, cellulose synthase, ribulose, cellulose synthase COBRA, leucine-rich protein, NAC domain protein, pectin esterase, ABA-responsive element binding factor 1, and heat stress protein. Our results is a step forward the understanding of the molecular response of S. spontaneum under salinity stress, which will lead to the identification of genes and transcription factors as novel targets for salinity tolerance in sugarcane.

摘要

甜高粱是甘蔗的野生近缘植物,具有很强的耐旱性和耐盐性。开发耐盐性种质资源是印度的一个主要重点领域。在这项研究中,我们利用抑制性消减杂交(SSH)结合测序技术,鉴定了来自印度不同地理区域的甜高粱克隆在盐胁迫下表达上调的转录本。我们对 SSH 文库的测序结果表明,95%的转化体含有 200-1500bp 大小的插入片段。经过消减处理后,我们在盐处理的样品中鉴定出了 314 个差异表达的转录本,随后通过定量实时聚合酶链反应进行了验证。功能注释和途径分析表明,上调的转录本是蛋白质修饰、应激和激素信号以及细胞壁发育和木质素形成的结果。显著上调的转录本包括 UDP 葡萄糖脱氢酶、纤维素合酶、核酮糖、纤维素合酶 COBRA、富含亮氨酸的蛋白、NAC 结构域蛋白、果胶酯酶、ABA 反应元件结合因子 1 和热应激蛋白。我们的研究结果进一步了解了甜高粱在盐胁迫下的分子响应,这将有助于鉴定甘蔗耐盐性的基因和转录因子等新靶标。

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