Regulatory roles of Jatropha curcas transcription factor JcTCP2 in leaf morphogenesis
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摘要: TCP（TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR1/2）转录因子是一类植物特有的转录因子家族，可分为I类和II类两大亚族；其中II类TCP转录因子是重要的叶片发育调节因子，在调控叶片大小和形态方面起主导作用。采用CRISPR-Cas9基因编辑技术通过农杆菌介导的遗传转化方法获得了木本油料植物小桐子（Jatropha curcas）的II类TCP转录因子基因JcTCP2的突变体jctcp2植株，并对其进行了表型观察和比较转录组分析，揭示了小桐子JcTCP2在叶片形态建成方面的调控作用。结果表明：获得的3个突变体（jctcp2-6、jctcp2-8和jctcp2-34）植株的JcTCP2基因发生移码突变及编码的蛋白质序列长度缩短。与野生型小桐子相比，jctcp2突变体表现出明显的叶片形态学异常，其叶片边缘的叶裂位置向上卷曲；进一步的组织学观察分析显示，突变体叶片的栅栏组织和上表皮细胞数量增加、排列紧密。jctcp2突变体叶片与野生型叶片的比较转录组分析结果表明，生长素信号通路（JcAX22D-1、JcAX22D-2、JcARF5、JcGH3-1、JcSAUR14-1、JcSAUR14-2、JcSAUR14-3、JcSAUR32、JcSAUR48-1、JcSAUR48-2、JcSAUR52、JcSAUR76、JcWALK22和JcTAR2）、细胞周期相关基因（JcCYCD5;1、JcSMR3、JcSMR6、JcKPR7）和细胞伸长相关基因（JcEXPA1、JcEXPA8和JcEXPA15）以及卷叶相关基因（JcYABBY1、JcNGA1）可能参与JcTCP2调控叶片卷曲的过程。
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关键词:
- 小桐子 /
- JcTCP2 /
- CRISPR-Cas9 /
- 叶片卷曲
Abstract: TCP (TEOSINTE BRANCHED1/CYCLOIDEA/ PROLIFERATING CELL FACTOR1/2) transcription factors constitute a plant-specific family divided into two major subclades: Class I and Class II. Among these, Class II TCP transcription factors serve as pivotal regulators of leaf development, playing a dominant role in modulating leaf size and morphology. This study employed CRISPR-Cas9 gene editing technology to generate mutant plants (jctcp2) of the Class II TCP transcription factor gene JcTCP2 in the woody oilseed plant Jatropha curcas through Agrobacterium-mediated genetic transformation. Phenotypic observation and comparative transcriptome analysis were conducted, revealing the regulatory role of JcTCP2 in leaf morphogenesis in J. curcas. The main findings were as follows: Three mutant lines (jctcp2-6, jctcp2-8, and jctcp2-34) were obtained, in which the JcTCP2 protein underwent frameshift mutations resulting in truncated protein sequences. Compared to wild-type J. curcas, the jctcp2 mutants exhibited distinct leaf morphological abnormalities, showing upward curling at leaf margin serrations. Histological analysis further revealed that palisade tissue and upper epidermal cells were more densely packed in mutant leaves. Comparative transcriptome analysis of jctcp2 mutant leaves versus wild-type leaves revealed that the auxin signaling pathway (JcAX22D-1, JcAX22D-2, JcARF5, JcGH3-1, JcSAUR14-1, JcSAUR14-2, JcSAUR14-3, JcSAUR32, JcSAUR48-1, JcSAUR48-2, JcSAUR52, JcSAUR76, JcWALK22, and JcTAR2), cell cycle-related genes (JcCYCD5;1, JcSMR3, JcSMR6, JcKPR7), cell elongation-related genes (JcEXPA1, JcEXPA8, and JcEXPA15), and leaf-rolling-related genes (JcYABBY1, JcNGA1) may participate in the JcTCP2-mediated regulation of leaf curling.-
Key words:
- Jatropha curcas /
- JcTCP2 /
- CRISPR-Cas9 /
- leaf curl
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