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Mixing of maize and wheat genomic DNA by somatic hybridization in regenerated sterile maize plants

机译:通过体细胞杂交将玉米和小麦基因组DNA混合在再生的不育玉米植物中

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Intergeneric somatic hybridization was performed between albino maize (Zea mays L.) protoplasts and mesophyll protoplasts of wheat (Triticum aestivum L.) by polyethylene glycol (PEG) treatments. None of the parental protoplasts were able to produce green plants without fusion. The maize cells regenerated only rudimentary albino plantlets of limited viability, and the wheat mesophyll protoplasts were unable to divide. PEG-mediated fusion treatments resulted in hybrid cells with mixed cytoplasm. Six months after fusion green embryogenic calli were selected as putative hybrids. The first-regenerates were discovered as aborted embryos. Regeneration of intact, green, maize-like plants needed 6 months of further subcultures on hormone-free medium. These plants were sterile, although had both male and female flowers. The cytological analysis of cells from callus tissues and root tips revealed 56 chromosomes, but intact wheat chromosomes were not observed. Using total DNA from hybrid plants, three RAPD primer combinations produced bands resembling the wheat profile. Genomic in situ hybridization (GISH) using total wheat DNA as a probe revealed the presence of wheat DNA islands in the maize chromosomal background. The increased viability and the restored green color were the most-significant new traits as compared to the original maize parent. Other intermediate morphological traits of plants with hybrid origin were not found.
机译:通过聚乙二醇(PEG)处理,在白化玉米(Zea mays L.)原生质体和小麦(Triticum aestivum L.)的叶肉原生质体之间进行属间体细胞杂交。没有融合的亲本原生质体都不能产生绿色植物。玉米细胞仅能再生活力有限的基本白化苗,而小麦的叶肉原生质体却无法分裂。 PEG介导的融合处理产生具有混合细胞质的杂交细胞。融合后六个月,选择绿色胚性愈伤组织作为推定的杂种。发现第一个再生子是流产的胚胎。完整,绿色,玉米样植物的再生需要在无激素培养基上进行6个月的进一步传代培养。这些植物是不育的,尽管有雄花和雌花。愈伤组织和根尖细胞的细胞学分析显示56条染色体,但未观察到完整的小麦染色体。利用杂交植物的总DNA,三种RAPD引物组合产生了类似于小麦的条带。使用小麦总DNA作为探针的基因组原位杂交(GISH)表明,玉米染色体背景中存在小麦DNA岛。与原始玉米亲本相比,增加的生存力和恢复的绿色是最重要的新性状。没有发现具有杂种起源的植物的其他中间形态学特征。

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