CRISPR/Cas9诱导的尼古丁生物合成核心基因A622和BBL突变对烟草的影响:尼古丁含量减少和发育异常

IF 5.4 Q1 PLANT SCIENCES
Jin-hee Jeong , Eun-young Jeon , Young Jong Song , Min Ki Hwang , Yeongji Gwak , Jae-Yean Kim
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引用次数: 0

摘要

烟草(Nicotiana tabacum)因其具有精神活性的生物碱尼古丁而闻名,这给公共卫生带来了重大挑战。最近的研究将尼古丁生物合成的最后阶段与 BBL 和 A622 基因联系起来,但生物合成途径的这一部分在很大程度上仍未被探索,是我们认识过程中的一个 "黑箱"。在我们的研究中,我们采用了多靶点CRISPR/Cas9系统,以商业烟草品种弗吉尼亚的BBL和A622的同源基因为靶点,创建了各种突变体。这导致了植物发育和生物碱含量的显著变化。值得注意的是,外显子内含子边界缺失的突变株系a622a-38-5和a622l-3-9表现出植株高度和叶片数量的显著下降,以及生物碱(包括烟碱)的大幅减少。特别是 A622 家族的双突变体比六倍 BBL 突变体表现出更严重的影响,强调了 A622 在烟碱合成和植物发育中的独特作用。我们的研究结果表明,A622和BBL基因突变可大幅降低尼古丁和安他滨碱的含量,某些情况下可降低99.6%。这些结果凸显了基因组编辑在开发尼古丁含量显著降低的烟草品种方面的潜力。这项研究不仅加深了我们对尼古丁生物合成的了解,还为开发成瘾性更低的烟草产品提供了途径,从而为公共卫生事业做出了贡献。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Impact of CRISPR/Cas9-induced mutations in nicotine biosynthesis core genes A622 and BBL on tobacco: Reduction in nicotine content and developmental abnormalities

Tobacco (Nicotiana tabacum) is known for its psychoactive alkaloid nicotine, which presents significant public health challenges. Recent research has linked the final stages of nicotine biosynthesis with the BBL and A622 genes, yet this part of the biosynthetic pathway remains largely unexplored, representing a 'black box' in our understanding. In our study, we employed a multi-target CRISPR/Cas9 system to target homologous genes of BBL and A622 in commercial tobacco varieties Virginia, creating various mutants. This led to significant variations in plant development and alkaloid content. Notably, mutant lines a622a-38–5 and a622l-3–9 with exon-intron boundary deletions exhibited significantly decreased plant height and leaf number, along with a substantial reduction in alkaloids, including nicotine. Particularly, double mutants in the A622 family displayed more severe effects than sextuple BBL mutants, emphasizing the distinctive role of A622 in nicotine synthesis and plant development. Our findings demonstrate that mutations in A622 and BBL genes can drastically reduce nicotine and anatabine content, with some cases showing reductions up to 99.6%. These results underscore the potential of genome editing in developing tobacco varieties with significantly lower nicotine levels. This study not only enhances our understanding of nicotine biosynthesis but also contributes to public health efforts by providing a pathway to develop less addictive tobacco products.

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来源期刊
Current Plant Biology
Current Plant Biology Agricultural and Biological Sciences-Plant Science
CiteScore
10.90
自引率
1.90%
发文量
32
审稿时长
50 days
期刊介绍: Current Plant Biology aims to acknowledge and encourage interdisciplinary research in fundamental plant sciences with scope to address crop improvement, biodiversity, nutrition and human health. It publishes review articles, original research papers, method papers and short articles in plant research fields, such as systems biology, cell biology, genetics, epigenetics, mathematical modeling, signal transduction, plant-microbe interactions, synthetic biology, developmental biology, biochemistry, molecular biology, physiology, biotechnologies, bioinformatics and plant genomic resources.
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