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首页> 外文期刊>Plant Science: An International Journal of Experimental Plant Biology >Functional validation of Capsicum frutescens aminotransferase gene involved in vanillylamine biosynthesis using Agrobacterium mediated genetic transformation studies in Nicotiana tabacum and Capsicum frutescens calli cultures.
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Functional validation of Capsicum frutescens aminotransferase gene involved in vanillylamine biosynthesis using Agrobacterium mediated genetic transformation studies in Nicotiana tabacum and Capsicum frutescens calli cultures.

机译：使用农杆菌介导的烟草和辣椒愈伤组织培养中农杆菌介导的遗传转化研究，验证了香草醛生物合成中涉及的辣椒果实氨基转移酶基因的功能验证。

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摘要

Capsaicinoid biosynthesis involves the participation of two substrates viz. vanillylamine and C9-C11 fatty acid moieties. Vanillylamine which is a derivative of vanillin is synthesized through a transaminase reaction in the phenylpropanoid pathway of capsaicinoid synthesis. Here we report the functional validation of earlier reported putative aminotransferase gene for vanillylamine biosynthesis in heterologous system using Agrobacterium mediated genetic transformation studies in Nicotiana tabacum and Capsicum frutescens calli cultures. Molecular analysis tools comprising PCR and Southern blot analysis have shown the integration of the foreign gene in N. tabacum and C. frutescens calli cultures. The study shows the production of vanillylamine in transformed N. tabacum callus cultures and also the reduction of vanillylamine production when whole gene based antisense binary vector construct was used in transformation of C. frutescens callus cultures. Vanillylamine production, aminotransferase assay with Western blot analysis for crude proteins of transformants established the production of putative aminotransferase (pAMT) protein in alternate plant. The result is a clear evidence of involvement of the reported putative aminotransferase responsible for vanillylamine biosynthesis in capsaicinoid biosynthesis pathway, confirming the gene function through functional validation.
机译：辣椒素的生物合成涉及两种底物的参与。香草胺和C 9 -C 11 脂肪酸部分。香草醛衍生物香草醛胺是通过辣椒素合成的苯基丙烷途径中的转氨酶反应合成的。在这里，我们报告使用农杆菌介导的烟草和辣椒辣椒愈伤组织培养的农杆菌介导的遗传转化研究，较早报道的推定的氨基转移酶基因在异源系统中香草醛生物合成的功能验证。包括PCR和Southern印迹分析在内的分子分析工具表明，外源基因已整合到烟草和油状愈伤组织的愈伤组织中。该研究表明，当将基于全基因的反义二元载体构建体用于转化C. frutescens愈伤组织培养时，转化的烟草愈伤组织培养物中产生了香草醛胺，并且降低了香草胺的产生。香草醛胺的生产，用Western blot分析氨基转移酶的方法检测转化株的粗蛋白，从而确定了备用植物中推定的氨基转移酶（pAMT）蛋白的生产。结果清楚地证明了报道的负责香草醛生物合成的推定氨基转移酶参与了辣椒素生物合成途径，从而通过功能验证确认了基因功能。

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