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An efficient and cost effective method of RNA extraction from mucilage phenol and secondary metabolite rich bark tissue of tossa jute (C. olitorius L.) actively developing phloem fiber

机译:一种有效且经济有效的方法从活跃发育韧皮部纤维的黄麻(C. olitorius L.)的粘液富含苯酚和次生代谢产物的树皮组织中提取RNA。

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摘要

Tossa jute is an important natural fiber crop of Southeast Asian countries including India, Bangladesh, China, Thailand, Myanmar etc. Traditional industrial application of jute fiber is limited to the packaging products like hessians, sacks, etc. and the fiber found unsuitable for textile industries largely due to significantly high lignin content. Therefore, understanding genetic factors underlying lignin biosynthesis in tossa jute holds promise for jute based product diversification. The major limiting factor in undertaking such study is unavailability of efficient protocol for RNA extraction at secondary growth active stage of tossa jute. Here we report a simplified, swift and cost effective protocol for isolating fairly good quality RNA from bark tissue of 65-days-old field grown tossa jute plant with active secondary growth. The purity, concentration and integrity of extracted RNA ascertained. To confirm downstream amenability, isolated RNA samples were reverse transcribed and PCR analysis done by using CCoAMT1 primer. Results established that method of RNA extraction presented here is an improvement over the other methods, particularly for bark tissue of field grown tossa jute at advance developmental stage. Therefore, present study will enhance our ability to understand expression pattern of fiber formation and maturation related genes in mature bark tissue that holds key for much talked genetic manipulation of fiber quality via lignin optimisation in the crop.
机译:黄麻纤维是东南亚国家的重要天然纤维作物,包括印度,孟加拉国,中国,泰国,缅甸等。黄麻纤维的传统工业应用仅限于粗麻布,麻袋等包装产品,并且发现这种纤维不适合用于纺织工业主要是由于木质素含量很高。因此,了解黄麻木质素生物合成基础的遗传因素为基于黄麻的产品多样化提供了希望。进行此类研究的主要限制因素是无法获得在黄麻次生生长活跃期进行RNA提取的有效方案。在这里,我们报告了一种简化,快速且具有成本效益的方案,该方案可用于从65天大田间生长的tossa黄麻植物的树皮组织中分离出质量较好的RNA,该植物具有活跃的次级生长。确定提取RNA的纯度,浓度和完整性。为了确认下游适应性,将分离的RNA样品进行反转录,并使用CCoAMT1引物进行PCR分析。结果表明,本文介绍的RNA提取方法是对其他方法的改进,特别是对于在发育后期生长的黄麻大田的树皮组织。因此,本研究将增强我们理解成熟树皮组织中纤维形成和成熟相关基因的表达模式的能力,而成熟树皮组织中通过木质素最优化对纤维质量进行遗传控制的遗传控制至关重要。

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