| 引用本文: | 许 鹏, 谭秋锦, 宋海云, 郑树芳, 杨小州, 环秀菊,
张 涛, 周春衡, 韦媛荣, 王文林.澳洲坚果果仁MiMYB44L基因克隆与结构功能分析[J].广西植物,2024,44(12):2232-2241.[点击复制] |
| XU Peng, TAN Qiujin, SONG Haiyun, ZHENG Shufang, YANG Xiaozhou, HUAN Xiuju,
ZHANG Tao, ZHOU Chunheng, WEI Yuanrong, WANG Wenlin.Cloning, structural and function analysis of MiMYB44L gene in kernels of Macadamia integrifolia[J].Guihaia,2024,44(12):2232-2241.[点击复制] |
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| 澳洲坚果果仁MiMYB44L基因克隆与结构功能分析 |
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许 鹏, 谭秋锦, 宋海云, 郑树芳, 杨小州, 环秀菊,
张 涛, 周春衡, 韦媛荣, 王文林*
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广西南亚热带农业科学研究所, 广西 崇左 532400
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| 摘要: |
| 澳洲坚果(Macadamia integrifolia)是一种具有高经济价值的常绿坚果树,其果仁富含脂肪酸和蛋白质等营养成分。为鉴定澳洲坚果果仁中与营养成分形成的相关基因,该研究采用转录组学、基因克隆、荧光定量和生物信息学等技术从澳洲坚果果仁营养成分含量有显著差异的品种‘桂热1号'和‘A4'的果仁转录组中挖掘调控基因。结果表明:(1)转录组分析发现‘桂热1号'相比‘A4'果仁有上调基因1 667个,下调基因1 798个; KEGG富集分析发现差异基因主要在淀粉和糖代谢、氨基酸生物合成和碳代谢路径中。(2)发现一个差异表达基因gene-LOC122077931,其编码R2R3-MYB转录因子MYB44L,并采用RACE技术在‘桂热1号'果仁中克隆了MiMYB44L,其全长1 165 bp,ORF长度999 bp,编码332个氨基酸。(3)生物信息学技术证明MiMYB44L蛋白含有R2R3-MYB家族特征的SANT结构域,不含有信号肽和跨膜结构域,含有磷酸化位点。(4)测定了10个澳洲坚果品种果仁中蛋白质含量,发现MiMYB44L在澳洲坚果高蛋白质含量品种中的表达量显著高于低蛋白质含量品种,整体相关系数为0.54,达到极显著水平。该研究结果为深入解析MiMYB44L在澳洲坚果营养成分形成中的调控作用机制提供了理论指导。 |
| 关键词: 澳洲坚果, MiMYB44L, 果仁, 结构, 表达分析 |
| DOI：10.11931/guihaia.gxzw202401062 |
| 分类号:Q943 |
| 文章编号:1000-3142(2024)12-2232-10 |
| 基金项目:广西重点研发计划(桂科AB22035012); 崇左市科技计划项目(崇科20210710); 广西农科院基本科研业务费项目(桂农科2023YM46)。 |
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| Cloning, structural and function analysis of MiMYB44L gene in kernels of Macadamia integrifolia |
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XU Peng, TAN Qiujin, SONG Haiyun, ZHENG Shufang, YANG Xiaozhou, HUAN Xiuju,
ZHANG Tao, ZHOU Chunheng, WEI Yuanrong, WANG Wenlin*
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Guangxi South Subtropical Agricultural Science Research Institute, Chongzuo 532400, Guangxi, China
Guangxi South Subtropical Agricultural Science Research Institute, Chongzuo 532400, Guangxi, China
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| Abstract: |
| Macadamia nut(Macadamia integrifolia)is an evergreen nut tree with high economic value. Its kernel is rich in nutrients such as fatty acid and protein, etc. In order to further explore the main regulatory genes related to nutrient formation in M. integrifolia kernels, transcriptomics, gene cloning, fluorescence quantification PCR and bioinformatics techniques were used to screen potential regulatory genes from the kernel transcriptomes of ‘Guire No. 1' and ‘A4', which have significantly different nutrient contents in M. integrifolia kernels. The results were as follows:(1)Transcriptome analysis showed that 1 667 genes were up-regulated and 1 798 genes down-regulated in ‘Guire No. 1' kernel compared with those of ‘A4' kernel; KEGG enrichment analysis showed that the differential genes were mainly in starch and glucose metabolism, amino acid biosynthesis and carbon metabolism.(2)A significant differentially expressed gene gene-LOC122077931 encoding the R2R3-MYB transcription factor MYB44L was discovered. The MiMYB44L gene was cloned in kernels of ‘Guire No. 1' using RACE technology, which was 1 165 bp in length, 999 bp in ORF in length, and encoded 332 amino acids.(3)Bioinformatics analysis confirmed the presence of the SANT domain in the MiMYB44L protein, a hallmark feature of the R2R3-MYB family. The protein lacked both a signal peptide and a transmembrane domain but featured phosphorylation sites.(4)The protein contents in kernels of 10 M. integrifolia varieties were determined. And it was found that the expression of MiMYB44L gene in M. integrifolia varieties with high protein content was significantly higher than that in varieties with low protein content, and the overall correlation coefficient was 0.54, reaching a extremely significant level. The results of this study provide theoretical guidance for in-depth analysis of the regulatory mechanism of MiMYB44L gene in the formation of protein content in M. integrifolia. |
| Key words: Macadamia integrifolia, MiMYB44L, kernel, structure, expression analysis |
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