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园艺学报 ›› 2022, Vol. 49 ›› Issue (6): 1247-1260.doi: 10.16420/j.issn.0513-353x.2021-0323

• 研究论文 • 上一篇    下一篇

甜橙SWEET2a促进柑橘溃疡病菌侵染

麻明英1,2, 郝晨星1,2, 张凯1,2, 肖桂华1,2, 苏翰英1,2, 文康1, 邓子牛1,2, 马先锋1,2,*()   

  1. 1湖南农业大学园艺学院,国家柑橘改良中心长沙分中心,长沙 410128
    2教育部园艺作物种质创新与新品种选育工程研究中心,长沙 410128
  • 收稿日期:2021-12-10 修回日期:2022-02-28 出版日期:2022-06-25 发布日期:2022-07-04
  • 通讯作者: 马先锋 E-mail:maxf8006@126.com
  • 基金资助:
    国家重点研发计划项目(2018YFD10003000);湖南省自然科学基金项目(2020JJ4040)

CsSWEET2a Promotes the Infection of Xanthomonas citri subsp. citri

MA Mingying1,2, HAO Chenxing1,2, ZHANG Kai1,2, XIAO Guihua1,2, SU Hanying1,2, WEN Kang1, DENG Ziniu1,2, MA Xianfeng1,2,*()   

  1. 1National Center for Citrus Improvement Changsha,College of Horticulture,Hunan Agricultural University, Changsha 410128,China
    2Engineering Research Center for Horticultural Crop Germplasm Creation and New Variety Breeding,Ministry of Education,Changsha 410128,China
  • Received:2021-12-10 Revised:2022-02-28 Online:2022-06-25 Published:2022-07-04
  • Contact: MA Xianfeng E-mail:maxf8006@126.com

摘要:

柑橘溃疡病菌(Xanthomonas citri subsp. citri,Xcc)侵染感病种质冰糖橙叶片后,在侵染部位形成典型的火山口凸起症状,但在抗病种质枸橼C-05叶片侵染部位呈现逐渐褐色坏死,未形成溃疡病典型症状。基于前期冰糖橙和枸橼C-05叶片接种Xcc的转录组测序结果,分析鉴定的20个甜橙SWEETXcc诱导表达变化,结果显示SWEET2aSWEET17d在冰糖橙中受Xcc诱导上调表达,SWEET12b在枸橼C-05中受Xcc诱导上调表达;实时荧光定量PCR验证,SWEET2a在冰糖橙叶片中受Xcc诱导高表达而在枸橼C-05叶片中表达变化不显著;SWEET2a在不同柑橘种质同源基因编码的氨基酸序列相似性为94.6%,其启动子顺式作用元件在冰糖橙和枸橼C-05中的种类和数量存在较大差异。冰糖橙和枸橼C-05叶片瞬时过表达结果显示,SWEET2a促进Xcc的繁殖,且拟南芥过表达转基因株系接种毒性菌Pst.DC3000(Pseudomonas syringae tomato DC3000)后,单位叶面积菌含量显著高于野生型。SWEET2a蛋白部分定位于质膜,表明SWEET2a可能被Xcc劫持用于跨膜运输糖类物质至胞外,供给Xcc在细胞间隙定殖所需要的能量。

关键词: 柑橘, 柑橘溃疡病菌, Xcc, SWEET2a, 基因功能

Abstract:

After the infection of citrus canker pathogen Xanthomonas citri subsp. citriXcc),infected the leaves of the susceptible germplasm‘Bingtang’Sweet Orange,it formed typical crater bulge symptoms at the infected site,but gradually showed brown necrosis at the infected site of the leaves of the resistant germplasm Citron C-05,without typical symptoms of canker disease. Based on the transcriptome sequencing analysis of‘Bingtang’Sweet Orange and Citron C-05. leaves inoculated with Xcc,the expression of 20 CsSWEET induced by Xcc was compared. The results showed that SWEET2a and SWEET17d were up-regulated by Xcc in‘Bingtang’Sweet Orange,and SWEET12b was up-regulated by Xcc in Citron C-05;qRT-PCR verification only SWEET2a induced by Xcc high expression in ‘Bingtang’Sweet Orange and no significant change in Citron C-05,The amino acids sequence similarity of SWEET2a in different citrus germplasm homologs was 94.6%. The cis-acting elements of the promoter in‘Bingtang’Sweet Orange and Citron C-05 were significantly different in types and quantities. The transient overexpression of SWEET2a in‘Bingtang’Sweet Orange and Citron C-05 showed that SWEET2a promoted the reproduction of Xcc,and the transgenic Arabidopsis thaliana lines inoculated with the virulence strain Pst.DC3000(Pseudomonas syringae tomato DC3000)was significantly higher than that of the wild type. The SWEET2a protein is partially localized to the plasma membrane,suggesting that SWEET2a may be hijacked by Xcc for transmembrane transport of sugar extracellular to supply Xcc with energy needed for intercellular colonization.

Key words: citrus, Xcc, Citron C-05, SWEET2a, gene function

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