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利用 PacBio 长读长 RNA 测序技术揭示棉花在时间盐胁迫下转录组的变化及其调控机制。

Temporal salt stress-induced transcriptome alterations and regulatory mechanisms revealed by PacBio long-reads RNA sequencing in Gossypium hirsutum.

机构信息

College of Agriculture, Xinjiang Agricultural University, 311 Nongda East Road, Urumqi, 830052, P. R. China.

State Key Laboratory of Cotton Biology/Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture/Institute of Cotton Research of Chinese Academy of Agricultural Science, Anyang, 455000, Henan, China.

出版信息

BMC Genomics. 2020 Nov 27;21(1):838. doi: 10.1186/s12864-020-07260-z.

Abstract

BACKGROUND

Cotton (Gossypium hirsutum) is considered a fairly salt tolerant crop however, salinity can still cause significant economic losses by affecting the yield and deteriorating the fiber quality. We studied a salt-tolerant upland cotton cultivar under temporal salt stress to unfold the salt tolerance molecular mechanisms. Biochemical response to salt stress (400 mM) was measured at 0 h, 3 h, 12 h, 24 h and 48 h post stress intervals and single-molecule long-read sequencing technology from Pacific Biosciences (PacBio) combined with the unique molecular identifiers approach was used to identify differentially expressed genes (DEG).

RESULTS

Antioxidant enzymes including, catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) were found significantly induced under temporal salt stress, suggesting that reactive oxygen species scavenging antioxidant machinery is an essential component of salt tolerance mechanism in cotton. We identified a wealth of novel transcripts based on the PacBio long reads sequencing approach. Prolonged salt stress duration induces high number of DEGs. Significant numbers of DEGs were found under key terms related to stress pathways such as "response to oxidative stress", "response to salt stress", "response to water deprivation", "cation transport", "metal ion transport", "superoxide dismutase", and "reductase". Key DEGs related to hormone (abscisic acid, ethylene and jasmonic acid) biosynthesis, ion homeostasis (CBL-interacting serine/threonine-protein kinase genes, calcium-binding proteins, potassium transporter genes, potassium channel genes, sodium/hydrogen exchanger or antiporter genes), antioxidant activity (POD, SOD, CAT, glutathione reductase), transcription factors (myeloblastosis, WRKY, Apetala 2) and cell wall modification were found highly active in response to salt stress in cotton. Expression fold change of these DEGs showed both positive and negative responses, highlighting the complex nature of salt stress tolerance mechanisms in cotton.

CONCLUSION

Collectively, this study provides a good insight into the regulatory mechanism under salt stress in cotton and lays the foundation for further improvement of salt stress tolerance.

摘要

背景

棉花(Gossypium hirsutum)被认为是一种相对耐盐的作物,但盐分仍会通过影响产量和降低纤维质量而造成重大经济损失。本研究通过时间胁迫研究了一种耐盐陆地棉品种,以揭示其耐盐分子机制。在胁迫后 0h、3h、12h、24h 和 48h 时间点测量了盐胁迫(400mM)的生化响应,并采用太平洋生物科学公司(PacBio)的单分子长读测序技术和独特的分子标识符方法来鉴定差异表达基因(DEG)。

结果

在时间胁迫下,抗氧化酶(CAT、POD、SOD)被发现显著诱导,表明活性氧清除抗氧化机制是棉花耐盐机制的重要组成部分。基于 PacBio 长读测序方法,我们鉴定了大量的新转录本。延长盐胁迫持续时间会诱导大量 DEG。在与应激途径相关的关键术语下发现了大量的 DEG,如“对氧化应激的反应”、“对盐胁迫的反应”、“对水分胁迫的反应”、“阳离子转运”、“金属离子转运”、“超氧化物歧化酶”和“还原酶”。与激素(脱落酸、乙烯和茉莉酸)生物合成、离子稳态(CBL 相互作用丝氨酸/苏氨酸蛋白激酶基因、钙结合蛋白、钾转运体基因、钾通道基因、钠/氢交换体或反向转运体基因)、抗氧化活性(POD、SOD、CAT、谷胱甘肽还原酶)、转录因子(髓细胞瘤、WRKY、APETALA 2)和细胞壁修饰相关的关键 DEG 对棉花的盐胁迫反应非常活跃。这些 DEG 的表达倍数变化既有正响应,也有负响应,突出了棉花耐盐机制的复杂性。

结论

总之,本研究为棉花在盐胁迫下的调控机制提供了很好的见解,并为进一步提高棉花的耐盐性奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6b97/7694341/5f55f5a7fe8b/12864_2020_7260_Fig1_HTML.jpg

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