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“华心”叶片转录组分析揭示与长期冷胁迫相关的候选基因。

Transcriptomic Analyses of 'Huaxin' Leaf Reveal Candidate Genes Related to Long-Term Cold Stress.

机构信息

Key Laboratory of Cultivation and Protection for Non-wood Forest Trees of Ministry of Education and the Key Laboratory of Non-Wood Forest Products of Forestry Ministry, Central South University of Forestry and Technology, Changsha 410004, China.

Engineering Technology Research Center of Southern Hilly and Mountainous Ecological Non-Wood Forest Industry of Hunan Province, Changsha 410004, China.

出版信息

Int J Mol Sci. 2020 Jan 28;21(3):846. doi: 10.3390/ijms21030846.

Abstract

'Huaxin' is a new high-yielding timber cultivar of of high economic value, and has been widely cultivated in the red soil hilly region of Hunan Province of the People´s Republic of China in recent years. However, its quality and production are severely affected by low temperatures during flowering. To find genes related to cold tolerance and further explore new candidategenes for chilling-tolerance, Illumina NGS (Next Generation Sequencing) technology was used to perform transcriptomic analyses of . Huaxin' leaves under long-term cold stress. Nine cDNA libraries were sequenced, and 58.31 Gb high-quality clean reads were obtained with an average of 5.92 Gb reads for each sample. A total of 191,150 transcripts were obtained after assembly. Among them, 100,703 unigenes were generated, and 44,610 unigenes were annotated. In total, 1564 differentially expressed genes (DEGs) were identified both in the A_B and A_C gene sets. In the current study, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed, andrevealed a group of cold-responsive genes related to hormone regulation, photosynthesis, membrane systems, and osmoregulation; these genes encoded many key proteins in plant biological processes, such as serine/threonine-protein kinase (STPK), transcription factors (TFs), fatty acid desaturase (FAD), lipid-transfer proteins (LTPs), soluble sugars synthetases, and flavonoid biosynthetic enzymes. Some physiological indicators of . 'Huaxin' were determined under three temperature conditions, and the results were consistent with the molecular sequencing. In addition, the expression levels of 12 DEGs were verified using quantitative real-time polymerase chain reaction (qRT-PCR). In summary, the results of DEGs analysis together with qRT-PCR tests contribute to the understanding of cold tolerance and further exploring new candidate genes for chilling-tolerance in molecular breeding programs of 'Huaxin'.

摘要

“华鑫”是一个新的高产木材品种,具有很高的经济价值,近年来已在中华人民共和国湖南省的红壤丘陵地区广泛种植。然而,其品质和产量受到开花期低温的严重影响。为了寻找与耐寒性相关的基因,并进一步探索新的候选耐寒基因,我们利用 Illumina NGS(下一代测序)技术对长期冷胁迫下的“华鑫”叶片进行了转录组分析。共测序了 9 个 cDNA 文库,每个样本平均获得了 5.92Gb 的高质量清洁读数,共获得了 58.31Gb 的高质量清洁读数。组装后共获得 191150 条转录本。其中,有 100703 条 unigenes 被生成,44610 条 unigenes 被注释。共鉴定出 A_B 和 A_C 两组基因中 1564 个差异表达基因(DEGs)。在本研究中,进行了基因本体(GO)和京都基因与基因组百科全书(KEGG)通路富集分析,揭示了一组与激素调节、光合作用、膜系统和渗透调节相关的冷响应基因;这些基因编码了植物生物学过程中的许多关键蛋白,如丝氨酸/苏氨酸蛋白激酶(STPK)、转录因子(TFs)、脂肪酸去饱和酶(FAD)、脂质转移蛋白(LTPs)、可溶性糖合成酶和类黄酮生物合成酶。在三种温度条件下测定了“华鑫”的一些生理指标,结果与分子测序一致。此外,还使用定量实时聚合酶链反应(qRT-PCR)验证了 12 个 DEGs 的表达水平。总之,DEGs 分析结果与 qRT-PCR 测试一起有助于理解华鑫的耐寒性,并进一步探索新的候选耐寒基因,用于分子育种计划。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f84/7037897/c8f54162f6d0/ijms-21-00846-g001.jpg

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