青菜与芥菜花粉－体细胞原生质体融合的研究

STUDIES ON POLLEN-SOMATIC PROTOPLAST FUSION BETWEEN BRASSICA CHINENSIS AND B.JUNCEA

以聚乙二醇（ＰＥＧ）为诱导剂，试验用４种方法进行青菜（Ｂｒａｓｓｉｃａｃｈｉｎｅｎｓｉｓ）幼嫩花粉原生质体与芥菜（Ｂ．Ｊｕｎｃｅａ）下胚轴原生质体的融合，结果以修改的Ｔｅｒａｄａ等的方法最好。融合后原生质体的培养形成了愈伤组织，但未分化出芽，个别愈伤组织分化了根。用ＲＡＰＤ技术对任选的５个细胞系进行鉴定，发现１个细胞系含双亲的ＤＮＡ片段，可确认是青菜幼嫩花粉原生质体与芥菜下胚轴原生质体融合后产生的杂种细胞系。

The fusion between young plooen (mid-late unicellular to early bicellular) protoplasts of Brassica chinensis and hypocotyl protoplasts of B.juncea was induced by polyethylene glycol(PEG). The results showed that among four methods tried the modified Terada et al’s( 1987)method was the best. PEG- treated protoplasts divided and formed calli.A few calli differentiated into roots, but no shoots were regenerated. Arbitrarily selected 5 lines originated from PEG-treated protoplasts were analysed with random amplified polymorphic DNA(RAPD) markers,and one clone had a combination of both B. chinensis and B. juncea DNA band pattern. The conclusion is that this was a gameto-somatic hybrid clone resulted from the fusion between pollen protoplast of B.chinensis and hypocotyl protoplast of B. juncea.