清香桂红色与绿色茎干株系的RAPD差异分析

RAPD Difference Analysis between Red and Green Stem Plants of Sarcococca ruscifolia

【目的】分别以红色和绿色茎干的清香桂植株为材料,对其遗传物质进行分子水平上的差异检测。【方法】用混合DNA样品对200个10碱基随机引物进行筛选,然后用筛出的引物对红色和绿色茎干的清香桂单株DNA样品各14个进行RAPD-PCR扩增,对差异性条带进行回收测序和核苷酸序列分析。【结果】43个引物在28个样品中具有多态性,8个能在多数红色和绿色茎干植株中产生差异条带,其中3个引物(C06、M09和N10)可稳定地产生清晰的7个差异性条带,且条带C06-350在所有14个红茎植株类型中都出现而在14个绿茎植株中均缺失,其核苷酸序列长度为352～381 bp,在NCBI上未搜索到同源的核苷酸或氨基酸序列。【结论】红茎与绿茎清香桂植株之间发生了遗传变化,C06-350是与清香桂红茎特征紧密联系的1个标记。

[Purpose]The difference of total DNA between red stem plants and the green ones of Sarcococca ruscifolia was studied in order to understand the molecular basis controlling stem color.[Methods]The mixed DNA template was amplified with a total of 200 RAPD primers of 10 bp,and the primers which had polymorphic bands were screened by 14 red stem plants and 14 green ones.The differential bands were retrieve,sequenced and analysed.[Results]Forty-three of primers were polymorphic in 28 DNA samples,8 of which could produce different bands in most of red and green stem plants.Three specific primers,C06,M09 and N10,could stably produce 7 distinct bands,of which band C06-350 appeared in all red stem plants,but was absent from all green ones.The sequencing result of the C06-350 showed that the sequence size was 352-381 bp and no homologous nucleotide or amino acid sequence was matched in NCBI database.[Conclusion]The genetic material of the red stem type had changed indeed compared with the green one,and C06-350 was a marker closely related to the characteristics of the red stem.