混合小麦亲本与新麦草不对称体细胞杂交体系的建立

Asymmetric Somatic Hybridization Between Mixed Wheat and Psathyrostachys juncea

以小麦品种济南 1 77悬浮细胞系来源的原生质体与同品系胚性愈伤组织制备的原生质体混合后作为受体 ;以经过 380 μW cm2 紫外线照射 1min、2min的新麦草原生质体分别作为供体 ,用PEG法诱导融合。组合Ⅰ (1 76 +cha9+新麦草UV 1min)获得 1 6个再生克隆。经过形态学、同工酶、染色体和RAPD分析 ,确定其全部为属间体细胞杂种。其中的 5个克隆再生杂种植株。用 7对小麦SSR引物对杂种克隆的叶绿体基因组进行了分析 ;组合Ⅱ (1 76+cha9+新麦草UV 2min)只获得 3个克隆 ,且逐渐褐化死亡。表明以小麦济南 1 77的两种培养细胞混合作受体的融合体系有利于杂种的获得及再生 ;

Psathyrostachys juncea is a potential source of useful genes, such as the barley yellow dwarf virus resistance, salt tolerance and drought tolerance, for wheat improvement. Conventional sexual hybridization between wheat and Psathyrostachys juncea is very difficult to occur as the two are sexual incompatible. Somatic hybridization is a promising technique for creating hybrids across the sexual border. Here we report a fusion system for somatic hybridization of wheat using PEG method. Mixed protoplasts of two wheat (Triticum aestivum L. cv. Jinan 177) culture cells (cha9 and 176) were used as the recipients to fuse with the donors, the protoplasts of Psathyrostachys juncea (Fisch.) Nevski irradiated with ultraviolet light (UV) at an intensity of 380μW/cm 2 for 1 min or 2 min. Sixteen clones were generated in the combination Ⅰ, (wheat 176 + wheat cha9 + P. juncea 1 min UV treatment) and five of the hybrid clones could differentiate to green plants. All the regenerated clones were confirmed as somatic hybrids by cytological, isozyme, chromosome and random amplified polymorphic DNA (RAPD) analysis. Chloroplast genome of the hybrids was analyzed using 7 pairs of wheat specific chloroplast microsatellite (SSR) primers. Three clones were obtained from the combination Ⅱ(wheat 176 + wheat cha9 + P. juncea 2 min UV treatment), and all browned slowly and died in 3 months. This result indicated that the mixed wheat cells was helpful to the formation and regeneration of hybrid callus and the dosage of the UV had significant effect on the development of the fusion products.