Articles
TRANSFORMATION OF IAAM GENE INTO ANTHER-DERIVED EMBRYOS OF LOQUAT MEDIATED BY AGROBACTERIUM TUMEFACIENS
Article number
1092_6
Pages
49 – 53
Language
English
Abstract
The parthenocarpic gene iaaM was transferred into anther-derived embryos, using Agrobacterium-mediated transformation.
The results showed that 2-3-day pre-culture, 15-min infection and 3-day co-culture with 10 mg L-1 AS in the co-culture medium led to an increase in transformation efficiency.
Cefotaxime at 250 mg L-1 or Ampicillin at 750 mg L-1 were optimal.
Sixteen transgenic lines were obtained.
The iaaM gene-specific amplification results showed that 9 strains were amplified at the expected target bands, and the PCR positive rate was 56.3%. The PCR-Southern analysis indicated that 8 strains showed hybridization signal, which further proved that the iaaM gene was integrated into the genome of the embryos, and the transformation rate was about 0.5%.
The results showed that 2-3-day pre-culture, 15-min infection and 3-day co-culture with 10 mg L-1 AS in the co-culture medium led to an increase in transformation efficiency.
Cefotaxime at 250 mg L-1 or Ampicillin at 750 mg L-1 were optimal.
Sixteen transgenic lines were obtained.
The iaaM gene-specific amplification results showed that 9 strains were amplified at the expected target bands, and the PCR positive rate was 56.3%. The PCR-Southern analysis indicated that 8 strains showed hybridization signal, which further proved that the iaaM gene was integrated into the genome of the embryos, and the transformation rate was about 0.5%.
Publication
Authors
L. Tao, Y.Q. Wang, C.P. Pan, Z.W. Yang, Q.X. Deng, J. Liu
Keywords
culture media, Eriobotrya japonica, genetic transformation, growth regulator, parthenocarpy gene
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