金魏佳,何 佳,章燕如,范灵希,唐露静,叶佳燕,郑 颖,蒋 明.青花菜病程相关蛋白基因 BoPR2 的克隆与表达[J].广东农业科学,2018,45(12):76-81 |
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青花菜病程相关蛋白基因 BoPR2 的克隆与表达 |
Cloning and expression analysis of a pathogenesis-related protein gene BoPR2 from Brassica oleracea var. italica |
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DOI:10.16768/j.issn.1004-874X.2018.12.013 |
中文关键词: 青花菜 病程相关蛋白 BoPR2 克隆 表达分析 |
英文关键词: Brassica oleracea var. italica pathogenesis-related protein BoPR2 cloning expression analysis |
基金项目:浙江省大学生科技创新活动计划(新苗人才计划)项目(2017R430012);台州市科技计划项目
(162ny14);浙江省自然科学基金(LY19C150004) |
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中文摘要: |
病程相关蛋白基因以家族形式存在于植物中,在抗病反应中起着重要作用。以青花菜为试材,
利用 PCR法克隆到 1个PR2基因,定名为BoPR2。测序结果表明,BoPR2基因组全长 1 188 bp,具 1个内含子,
编码区全长 1 092 bp,编码 351 个氨基酸;序列比对和系统发育分析结果显示,BoPR2 与野甘蓝、甘蓝型
油菜和白菜 PR2 的相似度最高,在发育树上聚为一组,与亚麻荠、荠菜的相似度最低,亲缘关系最远;实
时定量 PCR 结果表明,BoPR2 的表达受芸薹根肿菌诱导,在 10 d 时的相对表达量最大,约为对照的 6 倍;
但 BoPR2 的表达不受核盘菌的诱导。BoPR2 基因的克隆与表达分析,为青花菜抗病机理研究及开展分子
育种奠定了基础。 |
英文摘要: |
Pathogenesis-related protein genes comprise a gene family in plants, and they play a key role in
disease resistance. In this study, a gene designated BoPR2 was isolated from broccoli material using the PCR method.
Sequencing results indicated that the full-length genomic DNA was 1 188 bp with an intron, and its coding sequence
was 1 092 bp encoding 351 amino acids. Sequence comparison and phylogenetic analysis results showed that higher
similarities were found between BoPR2 and PR2 proteins from Brassica oleracea var. oleracea and B. rapa subsp.
chinensis, and they were clustered into the same clade. The lowest similarities were seen between BoPR2 and PR2s
from both Camelina sativa and Capsella rubella, indicating their distinct relationships. Real-time quantitative PCR
results revealed that the expression of BoPR2 gene was induced by Plasmodiophora brassicae, and the highest relative
expression level was detected at 10 d after incubation with an approximately 6-fold increase. However, the expression
of BoPR2 was not affected by Sclerotinia sclerotiorum. Isolation and expression analysis of BoPR2 provide evidence for
further studies on disease resistance mechanism and broccoli molecular breeding. |
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