| 引用本文: | 朱云娜, 陈凤梅, 李芷娴, 王 斌, 冯慧敏, 胡 芳, 李海渤.芥菜BjGSTF12基因克隆及表达分析[J].广西植物,2024,44(2):267-280.[点击复制] |
| ZHU Yunna, CHEN Fengmei, LI Zhixian, WANG Bin,
FENG Huimin, HU Fang, LI Haibo.Cloning and expression analysis of BjGSTF12 in Brassica juncea[J].Guihaia,2024,44(2):267-280.[点击复制] |
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| 芥菜BjGSTF12基因克隆及表达分析 |
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朱云娜, 陈凤梅, 李芷娴, 王 斌, 冯慧敏, 胡 芳, 李海渤*
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广东省粤北食药资源利用与保护重点实验室/韶关学院 生物与农业学院, 广东 韶关 512005
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| 摘要: |
| 为了探究谷胱甘肽转移酶编码基因(GST)在芥菜花青素积累中的作用,该文以紫薹-绿薹芥菜近等位基因系为材料,克隆到1个花青素积累相关的GST基因,命名为BjGSTF12。该文对BjGSTF12编码蛋白及其启动子进行生物信息学分析,并分析其在绿薹、紫薹芥菜中的表达水平及其与花青素含量的关系。结果表明:(1)BjGSTF12的基因组和cDNA全长分别为808、651 bp,编码216个氨基酸,具有GST_N端和GST_C端保守结构域。然而,绿薹、紫薹芥菜BjGSTF12序列无区别。(2)BjGSTF12与拟南芥AtGSTF12亲缘关系最近,同属于φ亚家族。(3)2个芥菜品系BjGSTF12启动子序列存在4处碱基突变/插入,但二者顺式作用元件种类与数目相同,均含9个MYB结合位点、1个赤霉素响应元件、3个非生物胁迫响应元件。(4)紫薹芥菜花青素含量显著高于绿薹芥菜,BjGSTF12表达水平与花青素含量表现出类似变化规律。(5)互作蛋白网络分析表明,BjGSTF12与花青素合成关键酶、糖基化修饰、转运蛋白等蛋白存在互作。综上认为,BjGSTF12在芥菜薹茎花青素积累中可能发挥重要作用,BjGSTF12可能通过互作蛋白调控芥菜花青素合成、修饰、转运从而影响花青素积累。该文对深入研究GST在芥菜薹茎花青素积累的功能及作用机制奠定了一定理论基础。 |
| 关键词: 芥菜, GST, 生物信息学分析, 表达分析, 花青素积累 |
| DOI：10.11931/guihaia.gxzw202305039 |
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| 文章编号:1000-3142(2024)02-0267-15 |
| 基金项目:广东省科技专项资金项目(210903164532210); 韶关学院大学生创新创业计划项目(Sycxcy2022155)。 |
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| Cloning and expression analysis of BjGSTF12 in Brassica juncea |
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ZHU Yunna, CHEN Fengmei, LI Zhixian, WANG Bin,
FENG Huimin, HU Fang, LI Haibo*
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Guangdong Provincial Key Laboratory of Utilization and Conservation of Food and Medicinal Resources in Northern Region /
College of Biology and Agriculture, Shaoguan University, Shaoguan 512005, Guangdong, China
Guangdong Provincial Key Laboratory of Utilization and Conservation of Food and Medicinal Resources in Northern Region /
College of Biology and Agriculture, Shaoguan University, Shaoguan 512005, Guangdong, China
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| Abstract: |
| To explore the role of glutathione S-transferase gene(GST)in the accumulation of anthocyanin in Brassica juncea, one GST gene related to the anthocyanidin accumulation was cloned from near-isogenic lines of B. juncea with purple stalk and green stalk, and named as BjGSTF12. In this study, the bioinformatics characteristics of BjGSTF12 encoding protein and promoter were analyzed, the expression level of BjGSTF12 and the relationship with total anthocyanidin content were analyzed in B. juncea lines with purple stalk and green stalk. The results were as follow:(1)BjGSTF12 genes from B. juncea were successfully cloned, whose the full length of BjGSTF12 in genome and cDNA was 808 bp and 651 bp, encoding a protein of 216 amino acids. The BjGSTF12 contained the typical domains of GST proteins, namely GST_N and GST_C. However, their sequences of BjGSTF12 did not exhibit any differences between the two lines of B. juncea.(2)BjGSTF12 was closely related to AtGSTF12 in Arabidopsis, and both belonged to the φ subfamily of GST family.(3)The BjGSTF12 promoter sequences were cloned from two Brassica juncea strains of green stalk and purple stalk, and they exhibited four base mutations/insertions. However, the types and numbers of cis-acting elements did not show obvious differences between the two strains, including nine MYB transcription factor binding sites, one hormone response element, and three abiotic corresponding elements.(4)The total anthocyanidin content in B. juncea of purple stalk was significantly higher than that in green stalk ones, and the expression levels of BjGSTF12 in two lines were found to be similar to the total anthocyanidin content in both lines.(5)Protein interaction network analysis revealed that BjGSTF12 may interact with the key enzymes of anthocyanidin biosynthesis, glycosylation modification, and transporter proteins. In summary, BjGSTF12 is likely to play a key role in the accumulation of anthocyanidin in B. juncea by regulating its biosynthesis, modification, and transportation through interactions with other proteins. This study provides a theoretical reference for further study on the function of GST and the mechanism of anthocyanidin accumulation in B. juncea. |
| Key words: Brassica juncea, GST, bioinformatics analysis, expression analysis, anthocyanidin accumulation |
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