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《Acta Botanica Boreali-Occidentalia Sinica》 2011-02
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Genetic Transformation of phbA Gene into Arabidopsis thaliana Plastid and Pollen Phenotype of Transgenic Plants

YANG Zheng-an1,2,DING Yu-mei3,WU Xing-en1,CHI Shu-juan1,HE Yong-qin1,ZHANG Ying-hua1(1 Faculty of Horticulture and Landscape,Yunnan Agricultural University,Kunming 650201,China;2 Biotechnology and Germplasm Institute,Yunnan Academy of Agriculture Science,Kunming 650223,China;3 School of Horticulture and Landscape Architecture,Southwest University,Chongqing 400716,China)  
The most effective method in hybrid breeding is that many male sterility lines have been exploited for the production of F1 hybrid seeds.Based on the previously reported sequences of phbA gene,cloned from(Alcaligenes eutrophus) coding β-ketothiolase,we amplified phbA gene from plastid pBHR68 by PCR.After sequencing,digesting and ligasing,we substituted phbA gene for gfp gene in plastid vector pBio3-GFP,thus constructed plastid transformation vector pCTHBA that carries the expression cassette of prrn-phbA-aadA-TpsbA-ter.Then the vector was transformed into plastid of Arabidopsis thaliana using PDS-1000/He biolistic particle delivering system.We totally obtained 12 transgenic plants.The further PCR analysis fundamentally indicated that the phbA gene was intergrated into the plastid genome of A.thaliana.All 12 transgenic plants performed the male-sterile phenotype with 11 plants were lacking stamens and 1 plant failed to produce viable pollen by phenotype examination and microscopy scanning.
【Fund】: 国家自然科学基金(30972012);; 云南省自然科学基金(2007C205M 2008CD136 2008CD125)
【CateGory Index】: Q943.2
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