Identification and Validation of Expressed Sequence Tags from Pigeonpea (Cajanus cajan L.) Root.

International journal of plant genomics Pub Date : 2014-01-01 Epub Date: 2014-05-06 DOI:10.1155/2014/651912
Ravi Ranjan Kumar, Shailesh Yadav, Shourabh Joshi, Prithviraj P Bhandare, Vinod Kumar Patil, Pramod B Kulkarni, Swati Sonkawade, G R Naik
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Abstract

Pigeonpea (Cajanus cajan (L) Millsp.) is an important food legume crop of rain fed agriculture in the arid and semiarid tropics of the world. It has deep and extensive root system which serves a number of important physiological and metabolic functions in plant development and growth. In order to identify genes associated with pigeonpea root, ESTs were generated from the root tissues of pigeonpea (GRG-295 genotype) by normalized cDNA library. A total of 105 high quality ESTs were generated by sequencing of 250 random clones which resulted in 72 unigenes comprising 25 contigs and 47 singlets. The ESTs were assigned to 9 functional categories on the basis of their putative function. In order to validate the possible expression of transcripts, four genes, namely, S-adenosylmethionine synthetase, phosphoglycerate kinase, serine carboxypeptidase, and methionine aminopeptidase, were further analyzed by reverse transcriptase PCR. The possible role of the identified transcripts and their functions associated with root will also be a valuable resource for the functional genomics study in legume crop.

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鸽子豆(Cajanus cajan L.)根部表达序列标签的鉴定与验证
鸽子豆(Cajanus cajan (L) Millsp.)是世界干旱和半干旱热带地区雨养农业的重要食用豆类作物。它的根系深而广,在植物发育和生长过程中发挥着许多重要的生理和代谢功能。为了鉴定与鸽子豆根部相关的基因,研究人员通过归一化 cDNA 文库从鸽子豆(GRG-295 基因型)的根部组织中生成了ESTs。通过对 250 个随机克隆进行测序,共产生了 105 个高质量的 ESTs,其中 72 个单体由 25 个等位基因和 47 个单体组成。ESTs根据其推定功能被归入9个功能类别。为了验证转录本的可能表达,对四个基因,即 S-腺苷蛋氨酸合成酶、磷酸甘油酸激酶、丝氨酸羧肽酶和蛋氨酸氨肽酶,进一步进行了反转录酶 PCR 分析。已鉴定转录本的可能作用及其与根相关的功能也将成为豆科作物功能基因组学研究的宝贵资源。
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