棉花根系干旱响应序列标签的分离、鉴定及表达分析

June 1 Pub Date : 2020-03-25 DOI:10.36899/japs.2020.3.0088
Adil Jamal, M. N. Shahid, B. Aftab, A. Johargy, M. S. Alshmemri, B. Rashid, T. Husnain
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引用次数: 1

摘要

棉花是重要的经济纤维作物。季节性缺水和长期缺水会影响棉花产量。本研究旨在探讨干旱胁迫下棉花根系的转录组。采用渗透胁迫(5%重力湿度)处理棉花根部提取的mRNA构建cDNA文库。表达的序列标签提交到基因库EST数据库(JK757087-JK757798)进一步注释,预测同源性和功能。在104个e值小于1e -33的转录本中,已知同源物82个(78.84%),未知同源物22个(21.15%)。通过基因本体和KEGG分析,阐明了这些干旱响应est在生物调控、分子功能和细胞器等方面的关键作用。通过RT-qPCR验证了10个单基因在根和叶中的表达模式。这些单一基因包括WD重复序列、FRIGIDA、过氧化物酶、E3泛素连接酶、U-box结构域、RNA结合、钙环素结合、谷胱甘肽S转移酶、几丁质内质酶和金属硫氨酸样蛋白。从棉花根中成功构建了cDNA文库,并发现了几个关键的干旱响应转录本。本研究发现的新序列可为进一步探索其在干旱响应机制基因组学中的作用提供宝贵资源。
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ISOLATION, CHARACTERIZATION AND EXPRESSION ANALYSIS OF PUTATIVE DROUGHT RESPONSIVE EXPRESSED SEQUENCED TAGS FROM GOSSYPIUM ARBOREUM ROOTS
Cotton is an important economic fibre crop. Seasonal water shortages and long term water deficit can effect cotton yield. Current study aimed to explore the cotton root transcriptome under drought stress. mRNA extracted from cotton roots subjected to osmotic stress treatment (5% gravimetric humidity) was used to construct cDNA library. Expressed sequence tags submitted to gene bank EST database (JK757087-JK757798) further annotated to predict the homology and function. Total 104 transcripts with an E-value less than 1e -33 revealed 82 (78.84%) known homologs and 22 (21.15%) with uncharacterized proteins. Gene ontology and KEGG analysis of these drought responsive ESTs elucidated their key role in biological regulation, molecular functions and cellular organelles. Expression pattern of 10 unigenes were validated by RT-qPCR in roots and leaves. These unigenes included WD repeat, FRIGIDA, peroxidase, E3 ubiquitin ligase, U-box domain, RNA binding, calcyclin binding, Glutathione S transferase, endochitinase and metallothionine like protein. cDNA library was successfully constructed from cotton roots and revealed several key drought responsive transcripts. Novel sequences identified in this study can be valuable resource for further exploration studies to exploit their role in genomics of drought responsive mechanism.
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