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作物学报 ›› 2010, Vol. 36 ›› Issue (3): 517-525.doi: 10.3724/SP.J.1006.2010.00517

• 研究简报 • 上一篇    下一篇

甘蓝和白菜紫色酸性磷酸酶17基因家族的克隆和比较分析

卢坤1,2，张凯1,2,**，柴友荣1,2，陆俊杏1,2，唐章林1,2，李加纳1,2,*   

  1. 1西南大学农学与生物科技学院，重庆400716；2重庆市油菜工程技术研究中心，重庆400716
  • 收稿日期:2009-09-18 修回日期:2009-12-08 出版日期:2010-03-12 网络出版日期:2010-01-22
  • 通讯作者: 李加纳, E-mail: ljn1950@swu.edu.cn
  • 基金资助:

    本研究由国家高技术研究发展计划（863计划）项目（2006AA10Z1E6和2008AA10Z147），引进国际先进农业科学技术计划（948计划）项目（2006-Q04）和中国博士后科学基金资助项目（20090460717）资助。

Cloning and Comparative Analysis of PURPLE ACID PHOSPHATASE 17 Gene Families in Brassica oleracea and Brassica rapa

LU Kun1,2,ZHANG Kai1,2,**,CHAI You-Rong1,2,LU Jun-Xing1,2,TANG Zhang-Lin1,2,*   

  1. 1 College of Agronomy and Life Sciences, Southwest University, Chongqing 400716, China; 2 Chongqing Rapeseed Technology Research Center, Chongqing 400716, China
  • Received:2009-09-18 Revised:2009-12-08 Published:2010-03-12 Published online:2010-01-22
  • Contact: LI Jia-Na, E-mail: ljn1950@swu.edu.cn

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1686

被引次数

13

摘要：

从甘蓝和白菜中分别克隆到2个PAP17, 根据序列相似性, 将PAP17分为类型I (BoPAP17-1和BrPAP17-1)和类型II (BoPAP17-2和BrPAP17-2)。Southern杂交表明, 白菜和甘蓝中均只存在2个PAP17成员, 与克隆结果吻合。系统发育和分子进化分析表明, PAP17基因家族受纯化选择, 编码低分子量PAP蛋白。荧光定量PCR结果表明, PAP17在所检测的9个组织器官中均有表达, 尤以花和蕾中的表达量特别高, 种子中也有一定程度的表达, 表明其与体内储备态磷的动用有关, 尤其在花和蕾的发育阶段。磷饥饿诱导表达结果表明, 除BrPAP17-2在幼苗叶片中表达受抑制外, BrPAP17和BoPAP17在幼苗根部和叶片中均受磷饥饿诱导, 表达量先降后升, 诱导4 d或8 d后达到峰值; 恢复供磷4 d后, 表达量迅速降低至基础表达水平以下。与幼苗叶片相比, 白菜和甘蓝幼苗根部的PAP17磷饥饿诱导表达似乎更为强烈。这些结果表明PAP17在白菜和甘蓝根部可能参与了根外磷的活化与吸收及无机磷由根部向其他组织器官的转运。

关键词: 白菜, 甘蓝, 紫色酸性磷酸酶, 基因家族, 磷饥饿

Abstract:

Two PAP17 genes were cloned from parent species of Brassica napus, B. oleracea and B. rapa, respectively. According to sequence similarity, PAP17 genes could be divided into two types, type I (BoPAP17-1 and BrPAP17-1) and type II (BoPAP17-2 and BrPAP17-2). Southern hybridization resulted in two bands both in B. oleracea and B. rapa, this is accordance with former cloning results. Phytogenetic and molecular evolution analysis indicated that PAP17 genes in Brassica species underwent purifying selection, and their deduced proteins are typical low molecular weight PAP proteins. Expression patterns of BoPAP17 and BrPAP17 genes were assayed by fluorescent quantitative PCR. The results revealed that PAP17 genes expressed in all nine tested tissues and organs, with the extremely high expression in flower and bud, and certain expression in seeds at different stages, implying these PAP17 genes most likely mobilize phosphorus reserves in plants, particularly during flower and bud development stages. Under phosphate starvation conditions, expression of BrPAP17-2 in seedling leaf was restrained, while that of BrPAP17 and BoPAP17 in seedling root and leaf was induced, the expression levels declined in the first 24 hours, and then continuously increased with the maximal levels between four days and eight days after treatment. After four days of Pi-resupply, their expression declined below un-induced basal levels. In comparison with seedling leaf, it seems that BrPAP17 and BoPAP17 showed stronger phosphate starvation induced expression in seedling root. These results thus suggested that PAP17 genes in B. oleracea and B. rapa may be involved in external phosphorus assimilation and transferring inorganic phosphate from root to other tissues or organs.

Key words: Brassica rapa, brassica oleracea, Purple acid phosphatase, Gene family, Phosphate starvation


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