首页|灰毡毛忍冬MADS-box基因家族鉴定与CMB1基因克隆

灰毡毛忍冬MADS-box基因家族鉴定与CMB1基因克隆

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目的 鉴定灰毡毛忍冬Lonicera macranthoides MADS-box家族基因并进行生物信息学分析与表达模式验证,克隆湘蕾型和野生型灰毡毛忍冬MADS-box家族成员CMB1 全长.方法 基于转录组数据,利用在线工具对灰毡毛忍冬MADS-box进行生物信息学分析,并利用qRT-PCR验证MADS-box基因在不同品种中的表达模式,通过RT-PCR、RACE技术克隆湘蕾型和野生型灰毡毛忍冬CMB1基因全长.结果 28个灰毡毛忍冬MADS-box蛋白长度为89～359 aa,碱性蛋白占比85.7%,不稳定蛋白占比96.4%,亲水性蛋白占比 96.4%,均定位于细胞核,均为无跨膜结构的非分泌蛋白.转录组显示,与野生型比较,湘蕾型中 28个MADS-box基因有 17.86%表达下调,MIKCC型基因中有 18.75%表达下调.克隆得到在两个品种的灰毡毛忍冬花中高度特异性表达的CMB1基因全长,均包含一个738 bp的ORF,编码245个氨基酸.CMB1基因在两个品种的花中表达量存在显著差异(P<0.01),且与茎、叶相比,在花中高度特异性表达(P<0.01).结论 基于灰毡毛忍冬转录组数据,鉴定了 28个MADS-box家族基因,克隆得到湘蕾型与野生型灰毡毛忍冬CMB1 基因全长,为进一步研究灰毡毛忍冬花发育、优良表型形成的分子机制提供研究基础与理论依据.
Identification of the MADS-box gene family and cloning of the CMB1 gene in Lonicera macranthoides
Objective To identify the MADS-box gene family of Lonicera macranthoides,analyze its bioinformatics,validate the expression patterns,and clone the full-length sequences of CMB1,a member of the MADS-box family of Lonicera macranthoides of Xianglei and wild types.Methods Based on the transcriptome data,the bioinformatics of Lonicera macranthoides MADS-box was analyzed with online tools,the expression patterns of MADS-box genes in different cultivars were verified using qRT-PCR,and the full-length sequences of Lonicera macranthoides CMB1 gene of the Xianglei and wild types were cloned by RT-PCR and RACE techniques.Results The sequence lengths of 28 MADS-box proteins of Lonicera macranthoides ranged from 89 aa to 359 aa,with alkali proteins accounting for 85.7%,unstable proteins accounting for 96.4%,and hydrophilic proteins accounting for 96.4%,which were localized in the nucleus,and were non-secretory proteins without transmembrane structure.The transcriptome showed that compared with the wild type,17.86%of the 28 MADS-box genes in the Xianglei type and 18.75%of the genes of MIKCC type were downregulated.The full-length sequences of CMB1 genes,which were highly specifically expressed in two varieties of flowers of Lonicera macranthoides,were cloned.Both of them contained a 738 bp ORF,encoding 245 amino acids.There was a significant difference in expression level of CMB1 gene between the two varieties of flowers(P<0.01),and it was highly and specifically expressed in flowers compared to that in stems and leaves(P<0.01).Conclusion Based on the transcriptome data of Lonicera macranthoides,28 MADS-box family genes were identified,and the full-length sequences of CMB1 gene of Xianglei and wild types were cloned,which provides the research foundation and theoretical basis for the further study of molecular mechanism of development and superior phenotype formation of Lonicera macranthoides.

Lonicera macranthoidesMADS-box gene familyexpression analysisCMB1 genegene clonephenotypic variation

付学森、刘紫璇、王玲、龙雨青、曾娟、周日宝、刘湘丹

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湖南中医药大学药学院,湖南 长沙 410208

湘产大宗道地药材种质资源及规范化种植重点研究室,湖南 长沙 410208

湖南省普通高等学校中药现代化研究重点实验室,湖南 长沙 410208

湖南省中药饮片标准化及功能工程技术研究中心,湖南 长沙 410208

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灰毡毛忍冬 MADS-box基因家族 表达分析 CMB1基因 基因克隆 表型变异

国家自然科学基金项目湖南省自然科学基金项目湖南省自然科学基金项目国家现代农业产业技术体系湖南中医药大学研究生创新训练项目湖南中医药大学研究生创新训练项目2020年湖南省一流专业建设点湖南中医药大学重点学科中药学科

823739922021JJ304972021JJ30515CARS-212022CX792023CX152校行发规字[2023]2号

2024

10.3969/j.issn.1674-070X.2024.03.006

湖南中医药大学学报
湖南中医药大学

湖南中医药大学学报

CSTPCD
影响因子：1.098
ISSN：1674-070X
年,卷(期)：2024.44(3)
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