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无籽沙糖桔WUBE1基因的功能验证
李鹏1, 苗红霞2, 胡桂兵1, 秦永华1
1.华南农业大学园艺学院, 广州 510642;2.中国热带农业科学院热带生物技术研究所, 海口 571101
摘要:  为了解泛素活化酶E1基因(UBE1)在无籽沙糖桔自交不亲和反应中的作用,通过根癌农杆菌介导法将来源于自交不亲和无籽沙糖桔(Citrus reticulata ‘Wuzishatangju’)WUBE1基因转化烟草(Nicotiana tabacum)。结果表明,外源基因WUBE1已导入烟草基因组中并得到表达。转WUBE1基因的自交授粉组合花粉管在生长过程中,部分花粉管出现停止生长的现象,到达花柱基部的花粉管数量少于异交授粉和野生型自交组合。转WUBE1基因烟草的花粉生活力、发芽率、自交和异交后每个果荚中的种子数与野生型烟草无显著差异。这表明单一的WUBE1基因不能调控无籽沙糖桔自交不亲和反应,很可能是通过Ub/26S途径参与了无籽沙糖桔自交不亲和反应。
关键词:  无籽沙糖桔  自交不亲和  UBE1基因  烟草  遗传转化  功能
DOI:10.11926/jtsb.4002
分类号:
基金项目:国家自然科学基金项目(31471858,31000899);广州市科技计划项目(201607010314);广东省自然科学基金项目(S2013020013084,S2013010011950)资助
Functional Analyses of WUBE1 Gene Isolated from Citrus reticulata ‘Wuzishatangju’
LI Peng1, MIAO Hong-xia2, HU Gui-bing1, QIN Yong-hua1
1.College of Horticulture, South China Agricultural University, Guangzhou 510642, China;2.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China
Abstract:  In order to understand the role of UBE1 gene in self-incompatibility (SI) response of Citrus reticulata ‘Wuzishatangju’, a WUBE1 gene from the self-incompatible cultivar 'Wuzishatangju' was transferred into tobacco (Nicotiana tabacum) mediated by Agrobacterium tumefaciens. The results showed that the WUBE1 gene was integrated into genome and expressed in transgenic tobacco. Some pollen tubes of self-pollinated transgenic tobacco with WUBE1 became twisted and could not enter the embryo sac. The number of pollen tube entering into ovules in self-pollinated WUBE1 tobacco was fewer than that in wild type (WT) and cross-pollination (transgenic tobacco×WT). However, there was no significant difference in pollen viability, germination rate and seed number per pod between transgenic tobacco and WT. Therefore, it was suggested that only the WUBE1 could not regulate SI response of ‘Wuzishatangju’, and it was possible to form a complex in Ub/26S pathway involved in the SI reaction.
Key words:  Citrus reticulata  Self-incompatibility  UBE1 gene  Tobacco  Genetic transformation  Function

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