根癌农杆菌介导大果西番莲基因转化

以大果西番莲种子无菌苗的子叶和下胚轴为外植体,在添加不同质量浓度BA和IAA的MS系列培养基上进行不定芽的诱导,确定MS+BA2.0mg·L-1+IAA0.1 mg·L-1为最适的不定芽诱导培养基,下胚轴、子叶外植体诱导芽分化率分别为88.33％,90.00％。将诱导出的不定芽接种在添加不同质量浓度IBA和NAA的MS系列培养基上,进行不定根的诱导,确定MS+NAA0.5mg·L-1+IBA0.2mg·L-1为最适的不定根诱导培养基,生根率达92.86％。用三亲交配法将质粒pBICr先转移到E.coli DH5α,然后转移到根癌农杆菌。利用含有NPT-Ⅱ基因和CaMV35S启动子控制下的黄瓜花叶病毒外壳蛋白（CMV-CP）反义基因的表达载体,通过根癌农杆菌介导法将其导入西番莲受体细胞,在质量浓度为50mg·L-1的Kan+选择压力下培养,获得大果西番莲CP转基因再生植株。 

Establishment of Transgenic and Regeneration System of Giant Granadilla(Passiflora quadrangularis L.)

The establishment of regeneration system of Passiflora quadrangularis from cotyledon and hypocoty explants and the transformation of this plant with reverse CMV-CP gene are described in this paper. MS based medium containing 2 mg. L-BA and 0.1 mg.L-'IAA was confirmed the best medium for inducing shoots, and the medium of MS +0.5 mg.L"NAA 0.2 mg.L-IBA for rooting. The regeneration frequency reached 88.33 %~90.00% using this system. BI121 based expressive vector pBICr containing NPT-lI gene cassette as well as anti-CMVCP sequence controlled by CaMV 35S promoter and NOS terminator, was transformed into P. quadrangularis mediated by Agro-bacterium tumefaciens. Putative transgenic plantlets were screened on kanamycin medium and confirmed by dot blot hybridization.