【目的】分析望春玉兰基因组微卫星序列和分布特征,开发多态性高和稳定性好的SSR引物,并构建玉兰商业品种指纹图,为玉兰属植物群体遗传结构和遗传多样性研究提供分子标记资源,也为玉兰优良品种的遗传真实性鉴别和育种工作者知识产权保护提供技术支撑和科学依据。【方法】基于已公布的望春玉兰基因组序列信息,采用MISA软件对全基因组微卫星序列进行查找和分析,使用Primer Premier 5.0软件设计SSR引物。随机合成203对引物,利用6株不同望春玉兰的基因组DNA进行筛选,并进一步利用筛选出的多态性SSR引物构建26个玉兰商业品种的DNA指纹图谱。【结果】望春玉兰基因组中共检测出2820303个SSR位点,6碱基重复类型占比最高(62.13%),其次是2碱基(12.02%)和单碱基重复(9.98%);共发现501种重复基序,其中出现频率高的基序为AAACCT/AGGTTT(16.46%),其次是A/T(8.63%)和AG/CT(5.9%)。在随机合成的203对引物中,有94对(46.31%)能够对6株不同望春玉兰的基因组DNA进行有效扩增,有25对引物(12.32%)的扩增谱带清晰、易于判读、多态性高,多态信息量(PIC)在0.24~0.72之间。利用PIC值最高的前10对引物构建了26个玉兰商业品种的DNA指纹图谱数据库,共扩增出85种基因型,每对引物产生的基因型在4~18之间;引物鉴别力在0~13之间,最少采用3对引物即可将26个玉兰品种完全区分开。【结论】本研究在望春玉兰全基因组微卫星序列统计和分析的基础上,开发了10对条带清晰、多态性高、重复性好的SSR引物,并利用这些引物构建了26个玉兰商业品种的DNA指纹图谱,DNA指纹图比对结果显示本研究所涉及的26个玉兰品种不存在同种异名或异种同名现象。
The fiber length trait （FLT） of 538 individuals from nature birch population in Maorshan region, Heilongjang, China were measured, of which 100 individuals were selected as representative variety of correlated fragm...
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The fiber length trait （FLT） of 538 individuals from nature birch population in Maorshan region, Heilongjang, China were measured, of which 100 individuals were selected as representative variety of correlated fragments screening with random amplified polymorphism DNA （RAPD） technique. In total of 20 RAPD primers were tested through multiple regression analysis between amplified strip and the character behaviors, and a correlative segment BFLR-16 was obtained. The correlation coefficient between BFLI-16 and FLT was 0.6144, with the significant level of 1%. BFLI-16 was then cloned, sequenced and transformed into SCAR marker. The percentage of identifying long fiber birches by this SCAR was more than 92. The result indicates that the SCAR markers has high specificity for the long fiber individuals and is highly linked with the gene controlling the character of fiber length, and its existence is significantly correlative with the increase in the fiber length.
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