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园艺学报 ›› 2021, Vol. 48 ›› Issue (2): 355-366.doi: 10.16420/j.issn.0513-353x.2020-0377

• 研究报告 • 上一篇    下一篇

石榴花器官发育相关基因PgWUSPgBEL1克隆及其时空表达分析

赵玉洁, 刘翠玉, 招雪晴, 汪钰莹, 闫明, 苑兆和*()   

  1. 南京林业大学南方现代林业协同创新中心,南京林业大学林学院,南京 210037
  • 收稿日期:2020-08-04 修回日期:2020-11-01 出版日期:2021-02-25 发布日期:2021-03-09
  • 通讯作者: 苑兆和 E-mail:zhyuan88@hotmail.com
  • 基金资助:
    国家自然科学基金项目(31901341);江苏省自然科学基金青年基金项目(BK20180768);南京林业大学高层次人才科研启动基金项目(GXL2014070);南京林业大学高层次人才科研启动基金项目(GXL2018032);江苏省高校优势学科建设工程项目(PAPD)

Cloning and Spatiotemporal Expression Analysis of PgWUS and PgBEL1 in Punica granatum

ZHAO Yujie, LIU Cuiyu, ZHAO Xueqing, WANG Yuying, YAN Ming, YUAN Zhaohe*()   

  1. College of Forestry,Nanjing Forestry University,Nanjing 210037,China
  • Received:2020-08-04 Revised:2020-11-01 Online:2021-02-25 Published:2021-03-09
  • Contact: YUAN Zhaohe E-mail:zhyuan88@hotmail.com

摘要:

在前期构建的‘泰山红’石榴(Punica granatum L.)基因组数据库的基础上,鉴定并采用同源克隆技术克隆得到PgWUSPgBEL1全长CDS(Coding sequence)序列。PgBEL1编码区全长为1 851 bp,PgWUS为936 bp,分别编码616个和311个氨基酸。蛋白序列比对及系统进化分析发现,PgWUS与甜瓜(Cucumis melo var. makuwa)、黄瓜(C. sativus)的WUS进化关系较近;PgBEL1和巨桉(Eucatyptus grandis)具有较高的相似性,聚在同一分支。PgBEL1和PgWUS行使功能区域定位在细胞核。实时荧光定量PCR分析表明,PgBEL1在花发育的P1 ~ P4阶段(花蕾纵径小于12.0 mm),两性花中的表达量高于功能性雄花(雌蕊败育);PgBEL1在叶片中的表达量最高,是花萼的3.2倍、茎段的1.2倍,茎尖中最低;雌蕊中PgBEL1是雄蕊的1.16倍。在石榴花发育的P2和P3阶段(花蕾纵径5.1 ~ 10.0 mm),PgWUS在两性花中的表达量高于功能性雄花;PgWUS在花萼中的表达量最低,茎段中最高;PgWUS在雌蕊中的表达量是雄蕊的1.5倍、茎尖的1.8倍。

关键词: 石榴, WUS, BEL1, 基因克隆, 表达分析

Abstract:

Based on the pomegranate genome database,PgWUS and PgBEL1,the homolog of WUS and BEL1 were identified and cloned by homologous cloning. The full-length CDS sequences of PgBEL1 and PgWUS were 1 851 bp and 936 bp,encoding 616 and 311 amino acids,respectively. The protein sequence multiple alignment and phylogenetic analysis suggested that PgWUS had the highest evolution relationship with WUS in Cucumis melo var. makuwa and C. sativus. There was high similarity of amino acid sequences in PgBEL1 and BEL1 of Eucatyptus grandis. Subcellular localization results showed that both PgBEL1 and PgWUS were located in the nucleus. Real-time quantitative PCR analysis suggested that the expression level of PgBEL1 in bisexual flowers was higher than that in functional male flowers at P1 P4 periods(bud vertical diameter < 12.0 mm). PgBEL1 was highly expressed in leaf,it was 3.2 times than that in calyx while 1.2 times than that in stem,respectively. PgBEL1 expression was low in shoot apex. The expression level of PgBEL1 in pistil was 1.16 times than that in stamen. At the P2 and P3 periods of pomegranate flower development that bud vertical diameter was 5.1-10.0 mm,the expression level of PgWUS in bisexual flowers was higher than that in functional male flowers. PgWUS expression was the lowest in calyx and was the highest in stem. The expression level of PgWUS in pistil was 1.5 times than that in stamen and 1.8 times than that in shoot apex. There was a significant difference in the expression levels of PgWUS in pistil and stamen.

Key words: pomegranate, WUS, BEL1, gene clone, expression analysis

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