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作物学报 ›› 2010, Vol. 36 ›› Issue (05): 779-787.doi: 10.3724/SP.J.1006.2010.00779

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦光敏色素基因TaPhyB3的克隆和表达分析

李壮1,2,马燕斌1,2,**,蔡应繁3,吴锁伟2,肖阳4,孟凡华2,付风铃1,黄玉碧1*,杨建平2,3,*   

  1. 1四川农业大学玉米研究所,四川雅安625014;2中国农业科学院作物科学研究所,北京100081;3重庆邮电大学生物信息学院,重庆400065;4中国农业科学院研究生院,北京100081
  • 收稿日期:2009-11-26 修回日期:2010-02-08 出版日期:2010-05-12 网络出版日期:2010-03-15
  • 通讯作者: 杨建平, E-mail: yangjianping@caas.net.cn, Tel: 010-82105859; 黄玉碧, E-mail: yubihuang@sohu.com, Tel: 0835-2882331
  • 基金资助:

    本研究由转基因生物新品种培育重大专项(2008ZX08009-003, 2008ZX08010-002), 国家高技术研究发展计划(863计划)项目(2008AA10Z121, 2006AA10Z1F2), 重庆市自然科学基金(CSTC2009BA1088), 中央级公益性科研院所基本科研业务费专项(2060302-2), 国家博士后科学基金(20080440462), 教育部归国人员启动费和农业部回国人员择优项目资助。

Cloning and Expression Analysis of TaPhyB3 in Triticum aestivum

LI Zhuang1,2, MA Yan-Bin1,2,**,CAI Ying-Fan3,WU Suo-Wei2,XIAO Yang4,MENG Fan-Hua2,FU Feng-Ling1,HUANG Yu-Bi1,*,YANGJian-Ping2,3,*   

  1. 1 Maize Research Institute, Sichuan Agricultural University, Ya’an 450002, China; 2 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 3 College of Bio-information, Chongqing University of Posts and Tele-communication, Chongqing 400065, China; 4 Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2009-11-26 Revised:2010-02-08 Published:2010-05-12 Published online:2010-03-15
  • Contact: YANG Jian-Ping,E-mail:yangjianping@caas.net.cn,Tel:010-82105859;HUANG Yu-Bi,E-mail:yubihuang@sohu.com,Tel: 0835-2882331

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2066

被引次数

5

摘要：

从小麦品种中国春中克隆了编码光敏色素基因B的脱辅基蛋白，将其定位于4D染色体的长臂上，并命名为TaPhyB3。TaPhyB3的开放阅读框为3 501 bp，编码一个具有1 165个氨基酸、128 kD的蛋白质。它与水稻、玉米和拟南芥在氨基酸水平上的一致性分别为93%、90%和73%。对不同光线处理7 d小麦植株表达的分析表明，TaPhyB3在黑暗中表达最低、在白光下的表达水平最高，在远红光、红光、蓝光和白光下的表达水平分别是黑暗中的2.2、7.7、7.4和37.3倍。组织特异性表达分析表明，TaPhyB3在小麦幼苗所有组织中都表达，叶片中的表达水平是根的11.4倍。推测TaPhyB3的表达水平与小麦的光形态建成的程度呈正相关。

关键词: 小麦, 光敏色素B, 基因克隆, 基因表达

Abstract:

As an important regulator in growth, development, and metabolic activities of plant, light is perceived by a variety of photoreceptors that control developmental processes, such as germination, photomorphogenesis, flowering, and senescence. Phytochromes play a pivotal role in plant adaptability to ambient environment. AtPhyB has been found to be involved in the response to red light. At present, PhyB genes have been cloned in various plants, and the expression patterns and functions of the gene family have been studied in Arabidopsis thaliana, rice (Oryza sativa L.), and maize (Zea mays L.), but not in wheat (Triticum aestivum L.). The objectives of this study were to clone the full length of PhyB and study its structure and expression under different lights. The full-length cDNA sequence of PhyB, encoding the apoproteinof phytochrome B, was cloned from wheat cultivar Chinese Spring. This gene is located on chromosome 4D and designated TaPhyB3. This gene possesses four extrons and three introns and the open reading frame TaPhyB3 is 3 501 bases in length, which encodes predicted protein of 1 166 amino acids. The conserved domains of PhyB gene family, i.e., DAF-DOMAIN, PHYTOCHROME REGION, PAS-A DOMAIN, PAS-B DOMAIN, HISTIDINE RELATED DOMAIN 1, and HISTIDINE RELATED DOMAIN 2, were also observed in the predicted protein sequence. The alignment analysis of amino acid sequence showed that TaPhyB3 shared 93% or 90% identity with the PHYBs of rice or maize, respectively, but only 73% with that of Arabidopsis. After treated with continuous darkness, far-red, red, blue, and white lights for 7 d, young seedlings of Chinese Spring were sampled for TaPhyB3 expression analysis using real-time reverse transcription polymerase chain reaction (RT-PCR). TaPhyB3 expression levels in the wheat seedlings under far-red, red, blue, and white lights were 2.2, 7.7, 7.4, and 37.3 times as high as that in seedlings under darkness. When exposed to white light for 90 d, the TaPhyB3 expression was detected in root, stem, leaf, and spike. However, the gene was mainly expressed in above-ground organs of wheat seedling, and TaPhyB3 expression level in leaf was 11.4 times as high as that in root. The expression level of TaPhyB3 is speculated to positively correlate with the degree of the seedling photomorphogenesis.

Key words: Triticum aestivum, Phytochrome B, Gene cloning, Gene expression

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