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绿豆象卵诱导的黑绿豆(绿豆)发育种子中的转录本动态变化。

Bruchid egg induced transcript dynamics in developing seeds of black gram (Vigna mungo).

作者信息

Baruah Indrani K, Panda Debashis, M V Jagadale, Das Deba Jit, Acharjee Sumita, Sen Priyabrata, Sarmah Bidyut Kumar

机构信息

DBT-AAU Centre, Assam Agricultural University, Jorhat, Assam, India.

Distributed Information Centre, Department of Agricultural Biotechnology, Assam Agricultural University, Jorhat, Assam, India.

出版信息

PLoS One. 2017 Apr 27;12(4):e0176337. doi: 10.1371/journal.pone.0176337. eCollection 2017.

Abstract

Black gram (Vigna mungo) seeds are a rich source of digestible proteins, however, during storage these seeds are severely damaged by bruchids (Callosobruchus spp.), reducing seed quality and yield losses. Most of the cultivated genotypes of black gram are susceptible to bruchids, however, few tolerant genotypes have also been identified but the mechanism of tolerance is poorly understood. We employed Suppression Subtractive Hybridization (SSH) to identify specifically, but rarely expressed bruchid egg induced genes in black gram. In this study, Suppression Subtractive Hybridization (SSH) library was constructed to study the genes involved in defense response in black gram against bruchid infestation. An EST library of 277 clones was obtained for further analyses. Based on CAP3 assembly, 134 unigenes were computationally annotated using Blast2GOPRO software. In all, 20 defense related genes were subject to quantitative PCR analysis (qPCR) out of which 12 genes showed up-regulation in developing seeds of the pods oviposited by bruchids. Few major defense genes like defensin, pathogenesis related protein (PR), lipoxygenase (LOX) showed high expression levels in the oviposited population when compared with the non-oviposited plants. This is the first report on defense related gene transcript dynamics during the bruchid-black gram interaction using SSH library. This library would be useful to clone defense related gene(s) such as defensin as represented in our library for crop improvement.

摘要

黑豆(绿豆)种子是易消化蛋白质的丰富来源,然而,在储存期间,这些种子会受到豆象(Callosobruchus spp.)的严重损害,从而降低种子质量并造成产量损失。大多数黑豆栽培基因型对豆象敏感,不过,也已鉴定出少数耐受基因型,但对其耐受机制了解甚少。我们采用抑制性消减杂交(SSH)技术来特异性鉴定黑豆中由豆象卵诱导但很少表达的基因。在本研究中,构建了抑制性消减杂交(SSH)文库,以研究黑豆中参与抵御豆象侵害的防御反应的基因。获得了一个包含277个克隆的EST文库用于进一步分析。基于CAP3组装,使用Blast2GOPRO软件对134个单基因进行了计算注释。总共对20个与防御相关的基因进行了定量PCR分析(qPCR),其中12个基因在被豆象产卵的豆荚发育种子中表现出上调。与未被产卵的植株相比,少数主要防御基因如防御素、病程相关蛋白(PR)、脂氧合酶(LOX)在被产卵群体中表达水平较高。这是首次利用SSH文库报道豆象与黑豆相互作用过程中与防御相关的基因转录动态。该文库将有助于克隆如我们文库中所代表的防御素等与防御相关的基因,以用于作物改良。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c85d/5407641/e3d64a489677/pone.0176337.g001.jpg

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