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园艺学报 ›› 2020, Vol. 47 ›› Issue (12): 2317-2330.doi: 10.16420/j.issn.0513-353x.2020-0422

• 研究论文 • 上一篇    下一篇

杧果MiPGP1和MiPGP2的表达分析及其对拟南芥不定根形成的影响

李运合1,2,*,刘 敏2,3,刘建东2,吴青松2   

  1. 1闽台特色园林植物福建省高校重点实验室,闽南师范大学生物科学与技术学院,福建漳州 363099;2农业部热带果树生物学重点实验室,中国热带农业科学院南亚热带作物研究所,广东湛江 524091;3海南大学园艺学院,海口 570228
  • 出版日期:2020-12-25 发布日期:2021-01-06
  • 基金资助:
    国家自然科学基金项目(31372053);闽南师范大学校长基金项目(KJ2020004)

Expression Analysis of MiPGP1 and MiPGP2 and Their Effects on Adventitious Root Formation in Arabidopsis

LI Yunhe1,2,*,LIU Min2,3,LIU Jiandong2,and WU Qingsong2   

  1. 1Key Laboratory of Landscape Plants with Fujian and Taiwan Characteristics,Fujian Province University,School of Biological Science and Biotechnology,Minnan Normal University,Zhangzhou,Fujian 363099,China;2Key Laboratory of Tropical FruitBiology,Ministry of Agriculture,South Subtropical Crop Research Institute,Chinese Academy of Tropical Agricultural Sciences,Zhanjiang,Guangdong 524091,China;3 College of Horticulture,Hainan University,Haikou 570228,China
  • Online:2020-12-25 Published:2021-01-06

摘要: 杧果子叶切段不定根仅在靠近胚轴端的切面上形成,而在远离胚轴端的切面上则没有不定根形成,前期研究表明生长素极性运输可能在这个过程中起着重要作用,但生长素极性运输载体基因PGP在其中的作用机制尚不清楚。以‘粤西1号’杧果(Mangifera indica L.‘Yuexi 1’)子叶切段为材料,采用RT-PCR结合RACE方法得到了2个生长素极性运输载体基因(MiPGP1和MiPGP2)的cDNA及基因组DNA全长。MiPGP1和MiPGP2的cDNA全长分别为4 650和4 107 bp,其开放读码框长度分别为4 092和3 851 bp,分别编码1 363和1 289个氨基酸;其基因组DNA全长分别为6 430和5 747 bp,从起始密码子到终止密码子的长度分别为6 082和5 633 bp。氨基酸序列多重比对及系统发育树结果显示MiPGP1和MiPGP2属于多重抗药性/磷酸糖蛋白家族。荧光定量PCR结果表明,MiPGP1和MiPGP2在杧果子叶切段不定根形成过程中,在近胚轴端(生根端)和远离胚轴端(非生根端)都有表达,0 h时生根端和非生根端表达量都比较低,培养6 h后二者的表达量都显著上调直到第10天不定根形成,但非生根端表达量始终高于生根端;吲哚丁酸(IBA)预处理后,MiPGP2的表达量明显上调, MiPGP1变化不大;2,3,5–三碘苯甲酸(TIBA)预处理后,MiPGP1和MiPGP2的表达量变化不大,仅在个别时间点受到抑制。转基因功能验证结果表明,过量表达MiPGP2显著促进拟南芥下胚轴不定根的形成。结果表明MiPGP2可能参与了杧果子叶切段不定根的形成。

关键词: 杧果, MiPGP1, MiPGP2, 表达分析, 过量表达, 拟南芥, 不定根

Abstract: Adventitious roots form only at the proximal cut surface(PCS)but not at the distal cut surface(DCS)of mango cotyledon segments,it was suggested that polar auxin transport play important role in this process but the role of multi-drug-resistant/P-glycoprotein(MDR/PGP)in this rooting was unclear. In this study,two mango MDR/PGP genes,designated as MiPGP1 and MiPGP2,were isolated from the cotyledon of mango(Mangifera indica L.‘Yuexi 1’)using RT-PCR and RACE. The full-length cDNA of MiPGP1 and MiPGP2 were 4 650 and 4 107 bp,with the open reading frames of 4 092和3 851 bp,which encode two putative proteins of 1 363 and和1 289 amino acids,respectively. The genomic DNA sequences of MiPGP1 and MiPGP2 were 6 430 and 5 747 bp,with the lengths of 6 082 and 5 633 bp,respectively,from the start codon to the terminator codon. Phylogenetic tree analysis indicated that MiPGP1 and MiPGP2 belong to plant PGP1 group. Quantitative real-time PCR showed that MiPGP1 and MiPGP2 expressed in PCS(where adventitious root formed)or DCS(no adventitious root formed)throughout the adventitious root formation period. Meanwhile,the relative expression of MiPGP1 and MiPGP2 was low at 0 d,but which was significantly up-regulated from 6 h until adventitious root appeared at 10 d,and the relative expression was always higher in DCS than that of PCS. However,the relative expression of MiPGP2 was significantly up-regulated by the pre-treatment with indole-3-butyric acid(IBA)but not MiPGP1. Pre-treatment with 2,3,5-triiodobenzoic acid(TIBA)inhibited the relative expression of MiPGP1 and MiPGP2 at some time points. In additon,the results of MiPGP1 and MiPGP2 overexpression in Arabidopsis plants showed that there was a significant increasment in adventitious root quantity in MiPGP2 transgenic line in hypocotyl. In conclusion,MiPGP2 might play an important role during the adventitious root formation of mango cotyledon segments.

Key words: Mangifera indica, MiPGP1, MiPGP2, expression analysis, overexpression, Arabidopsis, adventitious root

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