基于3个DNA序列的不同形态类型诸葛菜的遗传关系分析及后代性状观察

Genetic relationship of different morphological types of Orychophragmus violaceus based on three DNA sequences and character observation of their progenies

选择4种形态类型(花紫色-果实光滑、花紫色-果实密生毛、花白色-果实光滑和花白色-果实密生毛)诸葛菜〔Orychophragmus violaceus(L.) O.E.Schulz〕共11个单株为实验材料,在进行ITS、trnL-F和psbA-trnH序列分析的基础上,采用邻接法(NJ)构建系统发育树,并对各形态类型的后代性状进行观察和分析。结果表明,诸葛菜的ITS、trnL-F和psbA-trnH序列长度分别为704、767和216 bp,合并序列长度为1 687 bp,G+C含量为43.5%;共有7个变异位点和7个信息位点,变异率0.41%。4个形态类型诸葛菜的trnL-F序列完全一致,ITS和psbA-trnH序列略有差异。其中,花紫色和花白色的果实密生毛类型中各有1个单株的ITS序列第519位碱基为C,其他单株为T;花紫色和花白色的果实光滑类型中各有1个单株的psbA-trnH序列第68位至第73位碱基依次为CAAAAA,其他单株均为TTTTTG。NJ系统发育树显示,4个类型诸葛菜之间没有完全清晰的界限,亲缘关系很近。后代性状观察结果表明,诸葛菜白色花为特化现象,果实光滑和密生毛是不稳定的性状。研究结果支持将毛果诸葛菜(O.violaceus var.lasiocarpus Migo)并入诸葛菜的分类处理。

The eleven individuals belonging to four morphological types（purple petal and glabrous fruit,purple petal and hairy fruit,white petal and glabrous fruit,white petal and hairy fruit） of Orychophragmus violaceus（L.） O.E.Schulz were chosen as experimental materials,the NJ phylogenetic tree was constructed based on ITS,trnL-F and psbA-trnH sequence analysis,and the characters of their progenies were observed.The results show that the length of ITS,trnL-F and psbA-trnH sequences is 704,767 and 216 bp,respectively,and the combined length of the three sequences is 1 687 bp with a G＋C content of 43.5%.In which,there are seven variable sites and seven informative sites with a variation rate of 0.41%.The trnL-F sequences of the four morphological types are complete agreement,while ITS and psbA-trnH sequences have a little difference.The base of 519th site of ITS sequence of only one purple petal individual and one white petal individual in hairy fruit types is C,while the others are T.And the bases from 68th to 73th site of psbA-trnH sequence of only one purple petal individual and one white petal individual in glabrous fruit types are CAAAAA,while the others are TTTTTG.The NJ phylogenetic tree shows that there is not a clearly bound among the four morphological types of O.violaceus,so the four types have a close relationship.The observation results of progeny character show that the white petal of O.violaceus is a special phenomenon,and the character of glabrous or hairy fruit is unstable.The research results support the classification that O.violaceus var.lasiocarpus Migo can be incorporated into O.violaceus.