Transformation of Phomopsis perniciosa Mediated by Agrobacterium tumefaciens

doi:10.3969/j.issn.1000-7091.2013.04.040

Abstract

Abstract: We developed an Agrobacterium- mediated transformation system for P．perniciosa by using the－ conidia of strain LXS230101 as transformation recipients，A． tumefacien strain EHA105 carring plasmid pBIG3C har- boring the hygromycin B phosphotransferase gene (hph)．Successful transformation of P．perniciosa was performord and the highest effiency reached on 686 transformants per 1 × 10⁶spores．The optimal transformation conditions were that 1 × 10⁶spores per milliliter of P．perniciosa－ conidia suspension were co- cultured with Agrobacterium cells at 25 ℃ for 72 h，in the presence of Co- culture medium containing acetosyringone (AS) at 200 mol /mL．The trans- formants were verified by PCR amplification and by Southern blot analysis with the hph primers and probe，respec- tively．The results showed that all the detected transformants could be amplified the target bands and the T- DNA was inserted into the genome of P．perniciosa． In addition，the transformants were stable when grown on PDA medium without hygromycin for five times．

Key words: Phomopsis perniciosa, Genetic transformation, Transformation efficiency, Transformants identification

CLC Number:

S436.6

WANG Hai-yan, LI Bao-hua, LI Gui-fang, WANG Cai-xia. Transformation of Phomopsis perniciosa Mediated by Agrobacterium tumefaciens[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(4): 218-222. doi: 10.3969/j.issn.1000-7091.2013.04.040.

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