Reliable Production of Transgenic Cassava Plants Mediated by Agrobacterium tumefaciens
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Transformed cassava plants were successfully produced from friable embryogenic calli derived from leaf lobes byAgrobacterium tumefaciens-mediated transformation. We used A. tumefaciens strain LBA4404 containing pCAMmPHY whichcontained a binary vector with genes for β-glucuronidase (GUS) as a reporter and neomycin phosphotransferaseII (NPTI) forselection. Transformed calli and plantlets were selected on medium containing 20 or 30 mg l1 geneticin, respectively. At eachselection step, GUS activity in transformants resistant to geneticin was histochemically assayed. Southern blot analysis showed thatseven independent transgenic plants were identified and one to four copies of the transgenes was integrated into the casavagenome. RT-PCR confirmed GUS expression in the transgenic cassava. A leaf bleach assay by application of paromomycinshowed that the NPTII gene was functionally expressed in the transgenic cassava plants. FEC, genetic engineering, GUS, Manihot esculenta Crantz, NPTII, regeneration
Abstract
Transformed cassava plants were successfully produced from friable embryogenic calli derived from leaf lobes byAgrobacterium tumefaciens-mediated transformation. We used A. tumefaciens strain LBA4404 containing pCAMmPHY whichcontained a binary vector with genes for β-glucuronidase (GUS) as a reporter and neomycin phosphotransferaseII (NPTI) forselection. Transformed calli and plantlets were selected on medium containing 20 or 30 mg l1 geneticin, respectively. At eachselection step, GUS activity in transformants resistant to geneticin was histochemically assayed. Southern blot analysis showed thatseven independent transgenic plants were identified and one to four copies of the transgenes was integrated into the casavagenome. RT-PCR confirmed GUS expression in the transgenic cassava. A leaf bleach assay by application of paromomycinshowed that the NPTII gene was functionally expressed in the transgenic cassava plants. FEC, genetic engineering, GUS, Manihot esculenta Crantz, NPTII, regeneration