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园艺学报 ›› 2013, Vol. 40 ›› Issue (1): 69-78.

• 蔬菜 • 上一篇    下一篇

自交不亲和甘蓝亲和花粉授粉早期差异蛋白质分析

陈松, 曾静, 高启国, 朱利泉, 刘豫东, 任雪松, 王小佳   

  1. (1 西南大学农学与生物科技学院,重庆400716;2 西南大学园艺园林学院,南方山地园艺学教育部重点实验室,重庆400716)
  • 出版日期:2013-01-25 发布日期:2013-01-25

Differential Proteomic Analysis of Brassica oleracea Stigma and Pollen Proteins During the Early Stage of Compatible Pollination

 CHEN  Song, ZENG  Jing, GAO  Qi-Guo, ZHU  Li-Quan, LIU  Yu-Dong, REN  Xue-Song, WANG  Xiao-Jia   

  1. (1College of Agronomy and Biotechnology,Southweat University,Chonqging 400716,China; 2 College of Horticulture and Landscape Architecture,Southwest University,Key Laboratory of Horticulture Science for Southern Mountainous Regions,Ministry of Education,Chongqing 400716,China)
  • Online:2013-01-25 Published:2013-01-25

摘要: 为从蛋白质的角度揭示自交不亲和甘蓝亲和授粉早期花粉与柱头相互作用,将蛋白质组学方法应用于自交不亲和性甘蓝柱头与亲和花粉互作早期差异蛋白质的研究。结果表明,亲和授粉后3 ~ 5 min与1 h对比,有116个差异蛋白质点,而授粉后1 h与2 h差异不显著。按照特异表达与变化量大的蛋白质点优先原则,初步选取差异点中71个点做质谱分析,鉴定出31个可信差异蛋白质。与亲和授粉后3 ~ 5 min组相比,1 h组中特异表达蛋白质有19个,上调表达9个,下调表达3个。31种蛋白质的生物信息学分析表明,有两种蛋白与前人发现的与花粉管在柱头中生长发育相关蛋白相同,其他29种参与包括胁迫与防御应答在内多种生理过程。对早期差异蛋白中这种“既有促进花粉管发育的蛋白,也有与防御相关蛋白”现象分析表明,花粉管生长性侵入柱头的过程有可能伴随着柱头抗性的提高

关键词: 甘蓝, 授粉, 花粉—柱头相互作用, 蛋白质组学, 双向电泳

Abstract: In order to study the early pollen-stigma interaction in Brassica oleracea through proteomics analysis,inbred line A4 was pollinated with inbred line K101. Total stigma/pollen proteins were extracted by TCA/acetone at 3–5 min,1 h and 2 h after pollination,respectively. Total proteins were separated by two-dimensional gel electrophoresis and analyzed through MALDI-TOF-TOF-MS mass spectrometry. 116 proteins were differentially expressed contrasted the total stigma/pollen proteins of 1 h and 3–5 min after pollination,but only 7 proteins expressed differentially contrasted the total stigma/pollen proteins of 1 h and 2 h after pollination. 71 protein spots were analyzed through MALDI-TOF-TOF-MS mass spectrometry,and 31 proteins were identified homologous to known proteins by MASCOT analysis. Compared with total stigma/ pollen proteins of 3–5 min after pollination,19 of 31 identified proteins were specifically expressed 1 h after pollination,9 up-regulated and 3 down-regulated ones. The predicted functions of the 31 protein were related to pollen tube development,Ca2+ binding,biosynthetic process,ransmembrane transport,defense response,protein folding,carbohydrate and energy metabolism,regulation of translation and methylation. This study has documented the dynamics of protein expression during the early pollen-stigma interaction in Brassica oleracea and provides insights into the fundamental mechanisms involved in these processes.

Key words: Brassica oleracea L., pollination, pollen-stigma interaction, proteomics, 2-DE

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