Agrobacterium-mediated transformation of B. juncea reveals that BjuLKP2 functions in plant yellowing.

IF 4.4 1区 农林科学 Q1 AGRONOMY Theoretical and Applied Genetics Pub Date : 2024-08-09 DOI:10.1007/s00122-024-04707-9
Jing Zeng, Liang Zhao, Yuanqing Lu, Tonghong Zuo, Baowen Huang, Diandong Wang, Yawen Zhou, Zhongxin Lei, Yanling Mo, Yihua Liu, Jian Gao
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Abstract

Key message: A stable Agrobacterium-mediated transformation system was constructed for B. juncea, and BjuLKP2 was overexpressed, leading to plant yellowing. A stable and efficient transformation system is necessary to verify gene functions in plants. To establish an Agrobacterium-mediated transformation system for B. juncea, various factors, including the explant types, hormone combination and concentration, infection time and concentration, were optimized. Eventually, a reliable system was established, and two BjuLKP2 overexpression (OE) lines, which displayed yellowing of cotyledons, shoot tips, leaves and flower buds, as well as a decrease in total chlorophyll content, were generated. qRT-PCR assays revealed significant upregulation of five chlorophyll synthesis genes and downregulation of one gene in the BjuLKP2 OE line. Furthermore, antioxidant capacity assays revealed reduced activities of APX, CAT and SOD, while POD activity increased in the BjuLKP2 OE26. Additionally, the kinetic determination of chlorophyll fluorescence induction suggested a decrease in the photosynthetic ability of BjuLKP2 OE26. GUS assays revealed the expression of BjuLKP2 in various tissues, including the roots, hypocotyls, cotyledons, leaf vasculature, trichomes, sepals, petals, filaments, styles and stigma bases, but not in seeds. Scanning electron revealed alterations in chloroplast ultrastructure in both the sponge and palisade tissue. Collectively, these findings indicate that BjuLKP2 plays a role in plant yellowing through a reduction in chlorophyll content and changes in chloroplasts structure.

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农杆菌介导的君子兰转化揭示了 BjuLKP2 在植物黄化中的功能。
关键信息:构建了一个稳定的农杆菌介导的君子兰转化系统,BjuLKP2 被过量表达,导致植株黄化。要验证基因在植物中的功能,需要一个稳定高效的转化系统。为了建立农杆菌介导的君子兰转化系统,我们优化了各种因素,包括外植体类型、激素组合和浓度、感染时间和浓度。通过 qRT-PCR 检测发现,在 BjuLKP2 OE 株系中,五个叶绿素合成基因显著上调,一个基因下调。此外,抗氧化能力测定显示,BjuLKP2 OE26 的 APX、CAT 和 SOD 活性降低,而 POD 活性提高。此外,叶绿素荧光诱导的动力学测定表明,BjuLKP2 OE26 的光合能力下降。GUS 检测表明,BjuLKP2 在不同组织中都有表达,包括根、下胚轴、子叶、叶脉、毛状体、萼片、花瓣、花丝、花柱和柱头基部,但在种子中没有表达。扫描电子显微镜显示,海绵和栅栏组织的叶绿体超微结构都发生了改变。总之,这些研究结果表明,BjuLKP2 通过降低叶绿素含量和改变叶绿体结构在植物黄化中发挥作用。
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来源期刊
CiteScore
9.60
自引率
7.40%
发文量
241
审稿时长
2.3 months
期刊介绍: Theoretical and Applied Genetics publishes original research and review articles in all key areas of modern plant genetics, plant genomics and plant biotechnology. All work needs to have a clear genetic component and significant impact on plant breeding. Theoretical considerations are only accepted in combination with new experimental data and/or if they indicate a relevant application in plant genetics or breeding. Emphasizing the practical, the journal focuses on research into leading crop plants and articles presenting innovative approaches.
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