工业大麻EMS突变体及大麻素合成相关基因表达分析
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作者:
作者单位:

1.湖南科技职业学院药学院,长沙 410004;2.中国农业科学院麻类研究所,长沙 410205

作者简介:

研究方向为中药资源、天然药物,E-mail : ybliu2018@163.com

通讯作者:

张小雨,研究方向为作物种质资源学,E-mail : zhangxiaoyuyuyuyu@163.com

中图分类号:

基金项目:

湖南省自然科学基金部门联合基金(2023JJ60211);国家自然科学基金青年基金(32200219)


Construction of the EMS Mutagenesis Library for Industrial Hemp and Expression of Genes Related to Cannabinoid Biosynthesis
Author:
Affiliation:

1.School of Pharmacy, Hunan Vocational College of Science and Technology, Changsha 410004;2.Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205

Fund Project:

Foundation projects: Natural Science Foundation of Hunan Province, China(2023JJ60211); National Natural Science Foundation of China (32200219)

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    摘要:

    工业大麻(Cannabis sativa L.)是一种应用在造纸、医疗保健、纺织等领域的高价值经济作物。创制工业大麻新种质是开展品种选育和功能基因组研究的重要基础。本研究利用甲基磺酸乙酯(EMS)构建了中大麻资4号的突变体群体。通过设置8个EMS诱变剂浓度梯度处理,明确了0.8% EMS处理浓度对中大麻资4号诱变效果最好。对2000粒中大麻资4号种子进行诱变后,M1共成苗644株,萌发率为32.2%,其中包括叶色、花序和株高变异单株90株,突变率为13.98%。对M1突变体进行大麻二酚含量测定,获得3个高大麻二酚和2个低大麻二酚含量突变株;通过测序发现大麻二酚含量突变体中5个大麻素关键催化酶基因(CsPT4CsOAC1CsOLS1CsCBDASCsTHCA)及其启动子区域DNA序列均未发生突变,但是大麻二酚含量突变体中以上5个催化酶基因的表达水平较对照出现显著变化。由此推断,EMS处理导致了突变体中影响大麻二酚代谢途径的调控基因突变进而调控大麻二酚合成途径催化酶的表达,最终表现为突变体花头中大麻二酚含量显著变化。本研究通过创制EMS突变体群体为工业大麻育种提供了优异种质资源,也为进一步解析大麻素合成代谢途径的调控机制及挖掘相关基因提供良好材料和基础。

    Abstract:

    Industrial hemp (Cannabis sativa L.) is a high-value cash crop with applications in papermaking, healthcare, textiles, and other industries. The development of new industrial hemp germplasm is crucial for both breeding programs and functional genomic research. In this study, we constructed a mutant library of the variety ZHONGDAMAZI 4 (ZD4) using ethyl methanesulfonate (EMS) mutagenesis. By establishing a gradient of eight EMS concentrations, we determined that a 0.8% EMS concentration was optimal for inducing mutations in ZD4. After treating 2000 seeds with EMS solution, a total of 644 M1 seedlings were obtained, with a germination rate of 32.2%. Ninety individual plants exhibited variations in leaf color, inflorescence, and plant height, resulting in a mutation rate of 13.98%. High-performance liquid chromatography (HPLC) was used to measure the cannabidiol (CBD) content of the M1 plants, resulting in the identification of 3 high-CBD and 2 low-CBD mutants. Sequencing analysis didn’t reveal sequence variations in the coding and promoter sequences of five key cannabinoid biosynthetic enzyme genes (CsPT4CsOAC1CsOLS1CsCBDAS, and CsTHCA) in the CBD content mutants remained unchanged. However, the expression levels of these enzymes varied significantly compared to the control. It is inferred that EMS treatment induced mutations in regulatory genes affecting the CBD metabolic pathway in the mutants, thereby regulating the transcriptional expression and resulting in significant changes in CBD content. This study provides germplasm resources for industrial hemp breeding through the establishment of an EMS mutant library and lays a solid foundation for further elucidating the regulatory mechanisms of cannabinoid biosynthesis and related genes.

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刘永蓓,张小雨.工业大麻EMS突变体及大麻素合成相关基因表达分析[J].植物遗传资源学报,2025,26(2):394-403.

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  • 收稿日期:2024-04-30
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  • 在线发布日期: 2025-01-23
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