Development and application of PCR markers specific to the 1Ns chromosome of Psathyrostachys huashanica Keng with leaf rust resistance
Development and application of PCR markers specific to the 1Ns chromosome of Psathyrostachys huashanica Keng with leaf rust resistance
复制标题
DOI：
10.1007/s10681-014-1145-x
复制
发表时间：
2014-11-01
期刊：
影响因子：
1.9
通讯作者：
Chen, Xinhong
中科院分区：
文献类型：
--
作者：
Du, Wanli;Wang, Jing;Chen, Xinhong
利用基因组原位杂交(GISH)、表达序列标签-序列标签位点(EST-STS)和序列特征扩增区(SCAR)标记对小麦-华山新麦草二体附加系12-3进行了鉴定。有丝分裂和减数分裂GISH分析表明，该染色体含有42条小麦染色体和1对华山杨染色体。位于小麦同源第1组染色体上的8个EST-STS多位点标记在1NS二体附加系12-3中扩增出多态带，为华山杨所特有。此外，对随机扩增多态DNA标记OPAG10(986)和简单序列重复标记Xgwm601(135)的诊断片段进行了克隆、测序，分别转化为SCAR标记RHS153和SHS10，并用一系列不同的植物和一套小麦-P进行了验证。华山二体附加系(1NS-7 NS，2n=44=22 II)。结果表明，SCAR标记定位于华山松的NS基因组，并与1NS染色体连锁。此外，还对12-3进行了成虫期叶锈病抗性鉴定和农艺性状鉴定。这些新开发的EST-STS和SCAR标记将成为小麦育种者的强大工具，他们希望以低成本和高产量的方式筛选包含1NS染色体的基因型。
Wheat-Psathyrostachys huashanica Keng disomic addition line 12-3 was developed and characterized using genomic in situ hybridization (GISH), expressed sequence tag-sequence tagged site (EST-STS), and sequence characterized amplified region (SCAR) markers. Mitotic and meiotic GISH analyses indicated that it contained 42 wheat chromosomes and a pair of P. huashanica chromosomes. Eight EST-STS multiple-loci markers located on the homoeologous group 1 chromosomes of wheat amplified polymorphic bands in the 1Ns disomic addition line 12-3, which were unique to P. huashanica. These markers suggested that the introduced Ns chromosomes belonged to homoeologous group 1. Furthermore, diagnostic fragments of random amplified polymorphic DNA marker OPAG10(986) and simple sequence repeat marker Xgwm601 (135) were cloned, sequenced, and converted into SCAR markers, i.e., RHS153 and SHS10, respectively, which were validated using a range of distinct plant species and a complete set of wheat-P. huashanica disomic addition lines (1Ns-7Ns, 2n = 44 = 22 II). The results demonstrated that the SCAR markers targeted the Ns genome of P. huashanica and they were linked to the 1Ns chromosome. In addition, 12-3 was evaluated to test its leaf rust resistance in the adult stages and its agronomic traits. These newly developed EST-STS and SCAR markers will be powerful tools for wheat breeders who want to screen for genotypes containing the 1Ns chromosome, with low costs and high throughput.