用流式细胞仪和RAPD快速鉴定柑橘体细胞杂种

Rapid identification of Citrus somatic hybrids through flow cytometry and RAPD analysis

利用流式细胞仪和随机扩增多态DNA(RAPD)方法对获得的3例柑橘体细胞杂种进行鉴定,结果表明,两者相结合可快速有效地鉴定体细胞杂种.所检测的3个组合共9株再生植株,有8株是四倍体体细胞杂种,1株为二倍体叶肉亲本型杂种.体细胞杂种一般表现为具有双亲的特征带,且综合双亲的所有带,但在部分杂种中检测到了双亲都没有的新带,也发现有丢失亲本的特征带或共有带的现象,说明融合后染色体发生了重组和交换;有的引物只检测到叶肉亲本的特征带,根据柑橘叶肉细胞无论单独培养还是共培养均不能再生的事实,可推测其为体细胞杂种或二倍体叶肉亲本型胞质杂种.对这两种方法相结合用于体细胞杂种鉴定的可行性进行了讨论.

In this paper, a rapid and efficient method to identify the somatic hybrids of Citrus was demonstrated. With the combination of flow cytometric technique and random amplification polymorphism DNA （RAPD） markers, regeneration from three fusions were successfully identified. Among the tested nine somatic hybrid plants, eight were proved tetraploid, and the rest one was diploid. The somatic hybrids usually showed specific bands and combined all the bands of both parents. In combinations, some new bands were detected, and occasionally specific bands of parents were lost, which indicated that chromosome recombination and exchange happened after fusion. Additionally, in some combinations, specific bands from leaf parent were detected, which revealed the plants were of putative hybrid origin, because leaf protoplast cannot redifferentiate by both single culture and co-culture. The reliability of this strategy in somatic hybrid identification was also discussed.