摘要
用ISSR分子标记技术对4个不同色泽坛紫菜(Porphyra haitanensis)丝状体品系(分别为桔黄色,褐绿色,棕红色和紫色)及1个野生型对照丝状体品系(褐红色)进行了遗传分析,共筛选出11条引物,PCR反应得到了129个扩增位点,多态位点比率达88.4%,有效等位基因数为1.6369,期望杂合度为0.3609,Shannon多样性指数为0.5265;根据Nei方法计算了这5个坛紫菜丝状体品系间的遗传相似性系数和遗传距离,结果发现4个不同色泽丝状体间的平均遗传距离为0.5727,它们同野生型对照的平均遗传距离为0.6564,同时应用UPMGA法对它们进行了聚类分析。最后应用引物812扩增的4个多态性位点构建了这5个不同坛紫菜丝状体的DNA指纹图谱,使每一品系都具有独特的DNA指纹,并将其转化成了计算机可以识别的数码指纹,可以方便地应用于坛紫菜丝状体的种质鉴定中。结果表明ISSR分子标记技术可以作为坛紫菜丝状体种质鉴定的有效技术手段,提供快捷、准确的鉴定结果。
Parphyra haitanensi.s isthe most economically important seaweeds in south China. ISSR （intersimple sequence repeat） markers were applied to analyze the genetic variation among 4 different color＇ s P. haitanensis filament lines （saffron, green, brown and purple） and a wild-type line （maroon）. 11 ISSR primers （selected from 35 primers） gave rise to 129 discernible DNA bands of which 114 （88.4%） were polymorphic; Effective number of alleles （ Ne）, expected heterozygosity （h） and Shannon＇ s information index （I） were 1. 6369, 0. 3609 and 0.5265 respectively which were also calculated from the ISSR results by software popgene 1.32 ; Genetic similarity （GS） and genetic distances （GD） were calculated by using Nei＇ s matching coefficient, the average GD among the 4 different color＇ s filament lines is 0. 5727, and the average GD between the 4 different color＇ s filament lines and the wild-type filament line is 0. 6564, all these results indicating a considerable genetic variation among the 5 filament lines. UPGMA cluster analysis based on Nei＇ s genetic distances divided the 5 filament lines into three groups： the saffron and green in one group, the brown and purple in the other group and the wild-type line （maroon） in another group. Then we used the 4 polymorphic bands amplified from the primer 812 to construct the DNA fingerprints of the 5 P. haitanensis filament lines, in the constructed DNA fingerprints, each P. haitanensis filament line has its unique fingerprinting pattern and can be easily distinguished from each other. At last we transformed the DNA fingerprints into digital fingerprints which can be recognized by computer to predigest the variety identified process. All the results of this paper show that the ISSR marker is a useful technique in the variety identification of P. haitanensis, which can offer the correct results quicker than other molecular markers.
出处
《水产学报》
CAS
CSCD
北大核心
2007年第1期105-111,共7页
Journal of Fisheries of China
基金
国家自然科学基金(40676077)
福建省科技重大专项资助项目(2004NZ03)
关键词
坛紫菜
ISSR标记
遗传分析
种质鉴定
Porphyra haitanensis
ISSR marker
genetic analysis
germplasm identification
作者简介
谢潮添（1977-），男，福建龙岩人，讲师，博士，主要从事海藻分子生物学研究。E-mail：ctxie@126．com
通讯作者：陈昌生，E—mail：cschen@jmu．edu．cn
