摘要
以菊花品种‘秋水长流’DNA为试材,利用简并引物对菊花品种‘秋水长流’的DNA进行PCR扩增,对条带进行回收、克隆、测序,并通过生物信息学方法对获得的序列进行了碱基序列分析、同源性分析及聚类分析,以期为进一步探索反转录转座子在菊花遗传多样性形成中的作用奠定基础。结果表明:得到了一条230bp左右的目的条带。28条来自菊花的Ty1-copia反转录转座子反转录酶序列长度变化范围为207～232bp,GC与AT比例为1.06～1.63,同源性范围为25.3%～99.1%。其核苷酸序列经过系统聚类分析后可分为4个家族。所有氨基酸序列出现2～13个不同程度的终止密码子突变。并且将核苷酸序列与NCBI中已登录的不同物种Ty1-copia类反转录转座子反转录酶的核苷酸序列进行聚类分析。在菊花中得到4个家族的Ty1-copia类反转录转座子反转录酶序列在长度、碱基变化上的表现是不完全一致的,且序列同源性上存在多态性。系统进化树分析的结果表明,获得序列与其它植物中的Ty1-copia类反转录转座子反转录酶序列具有较高的同源性。菊花中的Ty1-copia类反转录转座子反转录酶序列间具有较高的异质性。
DNA of the chrysanthemum variety ‘Qiushui Changliu’was used as test material,and the DNA of the chrysanthemum variety‘Qiushui Changliu’was PCR amplified using degenerate primers.After the band was recovered,cloned,sequenced and analyzed by bioinformatics,and the sequence was analyzed by base sequence,homology analysis and cluster analysis.In order to further explore the function of retrotransposons in their genetic diversity and isolate Chrysanthemum Ty1-copia retrotransposons.The results showed that a target band of about 230 bp was obtain,the length of 28Ty1-copia retrotransposon reverse transcriptase sequences from chrysanthemums varied from 207-232 bp,the ratio of GC and AT was between 1.06 and 1.63,and the homology range was 25.3%-99.1%.These nucleotide sequences could be divided into 4families after systematic cluster analysis.After translation into amino acids,all sequences appeared to be 2-13 different degrees of termination codon mutation.Cluster and alignment analyses of the 28 sequences and other published sequences of the reverse transcriptase from other species in NCBI showed that four chrysanthemumfamilies of Ty1-copia retrotransposons reverse transcriptase sequences was obtained,and it was not completely consistent in length and base change,with diverse in the sequence homology.Phylogenetic tree analysis showed that it was highly homologous between the Ty1-copia retrotransposons reverse transcriptase sequences in the other plants and the obtained sequence.There existed a high heterogeneity in Chrysanthemum Ty1-copia retrotransposons reverse transcriptase sequences.
作者
王晓宇
王亚楠
李茫茫
李帅磊
丁红旭
王子成
WANG Xiaoyu;WANG Yanan;LI Mangmang;LI Shuailei;DING Hongxu;WANG Zicheng(College of Life Science,Henan University,Kaifeng,Henan 475004)
出处
《北方园艺》
CAS
北大核心
2019年第2期103-111,共9页
Northern Horticulture
基金
国家自然科学基金资助项目(31372090)
河南省自然科学基金资助项目(182300410026)
关键词
菊花
反转录转座子
反转录酶
RT基因
Chrysanthemum
reverse retrotransposon
reverse transcriptase
RT gene
