豇豆子叶节再生体系建立及Pti4基因遗传转化

Establishment of the Regeneration System of Cotyledonary Node and Genetic Transformation of Pti4 Gene in Cowpea

摘要:

为获得豇豆(Vigna unguiculata)遗传转化体系，以‘成豇七号’带一片子叶的子叶节作为外植体，对其高效再生体系和农杆菌介导抗病基因Pti4的遗传转化进行了研究。结果表明，豇豆无菌苗、不定芽诱导和不定芽伸长培养的最适培养基分别为MSB5 + 6-BA 3.0 mg L–1、MSB5 + 6-BA 1.0 mg L–1 + KT 0.06 mg L–1和MSB5 + 6-BA 0.5 mg L–1 + IBA 0.2 mg L–1。不定芽在MS培养基上能迅速诱导生根，获得完整植株。以豇豆子叶节为受体，通过农杆菌介导成功将 Pti4整合到‘成豇七号’抗性芽基因组中。因此，豇豆高效再生体系的建立为遗传育种研究奠定了基础。

Abstract:

In order to obtain genetic transformation system of cowpea (Vigna unguiculata), its cotyledon nodes with one cotyledon of ‘Chengjiang 7’ was used as explants, the regeneration system and Pti4 genetic transformation system by Agrobacterium-medium were studied. The results showed that the optimum mediums for aseptic seedling, adventitious bud introduction and adventitious bud elongation were MSB5 + 6-BA 3.0 mg L–1, MSB5 + 6-BA 1.0 mg L–1 + KT 0.06 mg L–1 and MSB5 + 6-BA 0.5 mg L–1 + IBA 0.2 mg L–1, respectively. Adventitious buds could rapidly rooting on MS medium to obtain full plantlets. The cotyledon node was used as receptor, the pit4 gene mediated by Agrobacterium was successfully transformed into the genome of resistant buds of ‘Chengjiang 7’. Therefore, the establishment of high efficient regeneration system for cowpea laid basis for studying on genetic transformation.