基于SSR分子标记的Nicotiana tobacum–N.plumbaginifolia异源染色体植株的鉴定与筛选  

Identification and Screening of Nicotiana tobacam–N. plumbaginifolia Heterologous Chromosome Plants Based on SSR Marker

作　　者：尚维 赵申清玉 党江波[1] 郭启高[1] 梁国鲁[1] 杨超 张艳 陈益银 SHANG Wei;ZHAO Shen-Qing-Yu;DANG Jiang-Bo;GUO Qi-Gao;LIANG Guo-Lu;YANG Chao;ZHANG Yan;CHEN Yi-Yin(College of Horticulture and Landscape,Southwest University,Chongqing 400716,China;Chongqing Tobacco Research Institute,Chongqing Municipal Tobacco Company,Chongqing 400716,China)

机构地区：[1]西南大学园艺园林学院,重庆400716 [2]中国烟草总公司重庆市公司烟草科学研究所,重庆400716

出　　处：《作物学报》2018年第11期1640-1649,共10页Acta Agronomica Sinica

基　　金：中国烟草总公司重庆市公司科技项目(NY20170401070001)资助~~

摘　　要：以烟草(Nicotiana tabacum)品种云烟87的八倍体(2n=8x=96)和野生烟草N. plumbaginifolia (2n=2x=20)的基因组DNA为模板,对340对烟草SSR引物进行筛选以获得能扩增多态性条带的引物。利用多态性引物对种间杂交后代及190株回交后代的基因组DNA进行扩增,并对N.plumbaginifolia中的SSR标记的连锁情况进行简要分析。经筛选获得了多态性引物29对。结果显示,在190株后代中, 159株的基因组DNA能扩增出N. plumbaginifolia的特异SSR位点,可以判定该159株为N. tabacum的N. plumbaginifolia异源染色体植株,其余31株植株可能不含有N. plumbaginifolia的染色体。经UPGMA聚类分析,本群体中植株的遗传多样性较为丰富,部分分子标记在后代中的出现具有完全相关性。29个标记中14个可确定来源于5条不同染色体,N.plumbaginifolia的29个位点在回交后代中的扩增效率并不相同,且效率均较低(低于31.00%),说明该杂种中N. plumbaginifolia基因组的垂直传递效率较低。利用SSR分子标记可以判定云烟87八倍体与N.plumbaginifolia杂交获得的后代为真杂种,且自该远缘杂种回交后代中筛选获得大量异源染色体植株。这些结果和筛选获得异源染色体植株为进一步创制N.tabacum-N.plumbaginifolia抗黑胫病单体附加系以及易位系奠定了基础。The 340 pairs SSR primers were amplified to select polymorphic primers amplifying polymorphic bands,in which the genomic DNA of the octoploid of Nicotiana tabacum Yunyan 87(2n=8x=96)and N.plumbaginifolia was extracted as a template.The polymorphic primers were used to amplify the genomic DNA of interspecific hybrids and 190 backcross progenies,and the linkage of SSR markers in N.plumbaginifolia was briefly analyzed.A total of 29 pairs of polymorphic primers.We found that N.plumbaginifolia specific SSR loci were amplified from 159 of the 190 BC1 plants.It was verified that 159 plants were N.tabacum– N.plumbaginifolia alien chromosome plants.By contrast,the other 31 plants might have no chromosomes of N.plumbaginifolia.Clustering analysis based on UPGMA indicated that the genetic diversity of the plants in BC1 population was relatively high,the appearance of some of the molecular markers in the offspring had a complete correlation.Fourteen in 29 markers could be identified from five chromosomes.The 29 N.plumbaginifolia specific loci were detected in BC1 plants in different rates,which were all lower than 31.00%,showing that vertical transmission rate of N.plumbaginifolia genome in the hybrid is low.The interspecies distant hybrids of Yunyan 87 octuploid(2n=8x=96)and N.plumbaginifolia could be identified as a true hybrid by using SSR marker,and a large number of N.tabacum–N.plumbaginifolia alien chromosome plants were screened out.All these results lay a foundation for production of black shank-resistant N.tabacum–N.plumbaginifolia monomer alien addition lines and alien translocations lines.

关 键 词：NICOTIANA tabacum N.plumbaginifolia SSR分子标记 异源染色体 黑胫病 

分 类 号：S572[农业科学—烟草工业]