通过不对称体细胞杂交向小麦转移青苗碱谷DNA

普通小麦"济南177"(Triticum aestivum L.cv.Jinan177)经继代培养和选择,形成了性质不同的愈伤组织,一种生长迅速,易于形成悬浮系,游离的原生质具有旺盛的分裂能力,但不能分化,称为Cha9;另一种具有一定能力,但游离原生质体分裂能力低,称为176.二者之一来源的原生质体与紫外线照射的青苗碱谷原生质体融合均不能获得再生植株.而将源于Cha9和176的两种原生质体混合,与经紫外线照射的青苗碱谷原生质体在PEG诱导下融合,融合产物再生了大量植株.再生的愈伤组织及植株经表现型、细胞学、有工酶、RAPD分析,证明了其杂种性质,用不麦叶绿体特异的简单得复序列(SSR)引物分析了再生杂种的叶绿体遗传组成情况.在不同的杂种克隆中,同时带有Cha9和176的遗传物质并含有供体核及胞质基因组的克隆4具有较高的分化能力,再生了大量生长旺盛的完整植株.

DNA Transfer from Wild Millet to Common Wheat by Asymmetric Somatic Hybridization

Following differing periods of long-term subculture and selection, two types of calli of the same wheat (Triticum aestivum L. cv. Jinan 177) were obtained. The one (named Cha 9) grew fast and easily formed cell suspensions that were non-regenerable, but the protoplasts possessed a high division capacity. The other (named 176) was regenerable, but the derived protoplasts grew slowly. Fusion combination between either Cha 9 or 176 protoplasts and UV-treated wild millet (Setaria italica L. Beaur.) protoplasts failed to produce regenerated green plants. However, when the two types of wheat protoplasts were mixed together as recipient and fused with wild millet, green plants were obtained. The hybrid nature of regener-ated calli and plants was confirmed by the analysis of cytological, isozyme, 5S rDNA spacer sequences and random amplified polymorphic DNA (RAPD). The chloroplast genomes of hybrids were analyzed with several wheat-specific chloroplast microsatellite (simple sequence repeat, SSR) primers. A hybrid clone carrying recipient DNA of Cha 9 and 176, as well as both nuclear and chloroplast donor DNA had a high regeneration capacity and produced more vigorous green plants than did the other clones.