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通过抑制性消减杂交鉴定与黄麻韧皮纤维发育相关的差异表达转录本。

Identification of differentially expressed transcripts associated with bast fibre development in Corchorus capsularis by suppression subtractive hybridization.

作者信息

Samanta Pradipta, Sadhukhan Sanjoy, Basu Asitava

机构信息

Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology, Kharagpur, 721302, India.

出版信息

Planta. 2015 Feb;241(2):371-85. doi: 10.1007/s00425-014-2187-y. Epub 2014 Oct 16.

DOI:10.1007/s00425-014-2187-y
PMID:25319611
Abstract

The present study documented the predominant role of WRKY transcription factor in controlling genes of different pathways related to fibre formation in jute and could be a candidate gene for the improvement of jute fiber. Understanding of molecular mechanism associated with bast fibre development is of immense significance to achieve desired improvement in jute (Corchorus sp.). Therefore, suppression subtractive hybridization was successfully applied to identify genes involved in fibre developmental process in jute. The subtracted library of normal Corchorus capsularis as tester with respect to its fibre-deficient mutant as driver resulted in 2,685 expressed sequence tags which were assumed to represent the differentially expressed genes between two genotypes. The identified expressed sequence tags were assembled and clustered into 225 contigs and 231 singletons. Among these 456 unigenes, 377 were classified into 15 different functional categories while others were of unknown functional category. Reverse Northern analysis of the unigenes showed distinct variation in hybridization intensity of 11 transcripts between two genotypes tested. The findings were also documented by Northern and real-time PCR analysis. Varied expression level of these transcripts suggested their crucial involvement in fibre development in this species. Among these transcripts, WRKY transcription factor was documented to be a most important transcript which was in agreement with its known role in other plant species in possible regulation related to cell wall biosynthesis, expansion and lignification. This report constitutes first systematic analysis of genes involved in fibre development process in jute. It may be suggested that the information generated in this study would be useful for genetic improvement of fibre traits in this plant species.

摘要

本研究记录了WRKY转录因子在控制黄麻纤维形成相关不同途径基因中的主导作用,其可能是用于改良黄麻纤维的候选基因。了解与韧皮纤维发育相关的分子机制对于实现黄麻(黄麻属)的理想改良具有极其重要的意义。因此,抑制性消减杂交技术被成功应用于鉴定参与黄麻纤维发育过程的基因。以正常圆果黄麻为测试方、其纤维缺陷型突变体为驱动方构建的消减文库,产生了2685个表达序列标签,这些标签被认为代表了两种基因型之间的差异表达基因。所鉴定的表达序列标签被组装并聚类为225个重叠群和231个单拷贝序列。在这456个单基因中,377个被归类为15个不同的功能类别,其他的功能类别未知。对这些单基因的反向Northern分析显示,在测试的两种基因型之间,11个转录本的杂交强度存在明显差异。Northern分析和实时PCR分析也证实了这些结果。这些转录本的表达水平各异,表明它们在该物种的纤维发育中起着关键作用。在这些转录本中,WRKY转录因子被证明是最重要的转录本,这与其在其他植物物种中可能参与细胞壁生物合成、扩张和木质化调控的已知作用相一致。本报告首次对参与黄麻纤维发育过程的基因进行了系统分析。可以认为,本研究产生的信息将有助于该植物物种纤维性状的遗传改良。

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本文引用的文献

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