Integration and expression of Xa21 in transgenic rice CX8621.

Lifen Gao; Pengcheng Liu; Zhihui Xia; Jiying Zhao; Jianan Shi; Guanghuai Jiang; Guozhen Liu; Wenxue Zhai

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Integration and expression of Xa21 in transgenic rice CX8621.

Author: Lifen GAO ¹ ; Pengcheng LIU ¹ ; Zhihui XIA ¹ ; Jiying ZHAO ¹ ; Jianan SHI ² ; Guanghuai JIANG ¹ ; Guozhen LIU ² ; Wenxue ZHAI ¹
Author Information

1. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
2. College of Life Sciences, Hebei Agricultural University, Baoding 071001, Hebei, China.
Publication Type:Journal Article
Keywords: T-DNA integration; Xa21; bacteria blight resistance; expression analysis; transgenic rice
MeSH: Animals; DNA Primers; Disease Resistance; genetics; Oryza; genetics; Plant Proteins; genetics; Plants, Genetically Modified; genetics; Polymerase Chain Reaction; Protein-Serine-Threonine Kinases; genetics; Seeds
From: Chinese Journal of Biotechnology 2016;32(9):1255-1263
CountryChina
Language:Chinese
Abstract: Agrobacterium tumefaciens-mediated transformation system has been widely applied. However, the function of target gene is affected by multiple factors. With this system, we obtained a transgenic rice line CX8621 carrying the bacterial blight resistance gene Xa21. In previous work, we have confirmed that it was selectable maker-free and vector backbone-free. And after 16 generations of breeding, it still maintained perfect resistance to bacterial blight disease. On this basis, we analyzed the integration and expression of Xa21 in CX8621 at the present study. First, based on the border sequences of plasmid pBXa21 and Xa21, we designed nested primers and assured the integrity of Xa21 in CX8621. Second, we cloned the flanking sequences and located Xa21 on chromosome 2 using improved Tail-PCR. Then we analyzed the expression pattern of Xa21 in several tissues and at different developmental stages by RT-PCR. The results show that Xa21 can be stably expressed in CX8621, agreeing well with the disease resistance response as reported previously. In addition, we detected the protein levels of XA21 in CX8621 with antibody of natural XA21 protein. Surprisingly, no XA21 protein was detected in the seeds of CX8621. Thus, the integration and expression analysis of Xa21 in CX8621 provided a part of scientific evidences for the safety assessment of genetically modified rice.