Isolation of the endosperm-specific LPAAT gene promoter from coconut (Cocos nucifera L.) and its functional analysis in transgenic rice plants
文献类型: 外文期刊
作者: Xu, Li 2 ; Ye, Rongjian 4 ; Zheng, Yusheng 1 ; Wang, Zhekui 1 ; Zhou, Peng 3 ; Lin, Yongjun 4 ; Li, Dongdong 1 ;
作者机构: 1.Hainan Univ, Minist Educ, Key Lab Trop Biol Resources, Haikou 570228, Hainan, Peoples R China
2.Chinese Acad Trop Agr Sci, Trop Crops Genet Resources Inst, Minist Agr, Key Lab Trop Crops Germplasm Resources Utilizat, Danzhou 571737, Hainan, Peoples R China
3.Chinese Acad Trop Agr Sci, Minist Agr, Inst Trop Biosci & Biotechnol, Key Lab Trop Crop Biotechnol, Danzhou 571737, Hainan, Peoples R China
4.Huazhong Agr Univ, Natl Key Lab Crop Genet Improvement, Wuhan 430070, Peoples R China
5.Huazhong Agr Univ, Natl Ctr Plant Gene Res, Wuhan 430070, Peoples R China
关键词: Coconut (Cocos nucifera L.);Endosperm-specific;Function analysis;Promoter;Transgenic rice
期刊名称:PLANT CELL REPORTS ( 影响因子:4.57; 五年影响因子:4.463 )
ISSN: 0721-7714
年卷期: 2010 年 29 卷 9 期
页码:
收录情况: SCI
摘要: As one of the key tropical crops, coconut (Cocos nucifera L.) is a member of the monocotyledonous family Aracaceae (Palmaceae). In this study, we amplified the upstream region of an endosperm-specific expression gene, Lysophosphatidyl acyltransferase (LPAAT), from the coconut genomic DNA by chromosome walking. In this sequence, we found several types of promoter-related elements including TATA-box, CAAT-box and Skn1-motif. In order to further examine its function, three different 5'-deletion fragments were inserted into pBI101.3, a plant expression vector harboring the LPAAT upstream sequence, leading to pBI101.3-L1, pBI101.3-L2 and pBI101.3-L3, respectively. We obtained transgenic plants of rice by Agrobacterium-mediated callus transformation and plant regeneration and detected the expression of gus gene by histochemical staining and fluorometric determination. We found that gus gene driven by the three deletion fragments was specifically expressed in the endosperm of rice seeds, but not in the empty vector of pBI101.3 and other tissues. The highest expression level of GUS was at 15 DAF in pBI101.3-L3 and pBI101.3-L2 transgenic lines, while the same level was detected at 10 DAF in pBI101.3-L1. The expression driven by the whole fragment was up to 1.76- and 2.8-fold higher than those driven by the -817 bp and -453 bp upstream fragments, and 10.7-fold higher than that driven by the vector without the promoter. Taken together, our results strongly suggest that these promoter fragments from coconut have a significant potential in genetically improving endosperm in main crops.
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