Liu Ai Feng, Wang Hong Gang, Hao Yuan Feng, Duan You Chen, Wang Yu Hai, Wu Xin Ru, Li Yan, Zhu Yu Li, Gao Ju Rong
Agronomy college of Shandong Agricultural University/Taian Subcentre of National Wheat Improvement Centre, Taian.
Fen Zi Xi Bao Sheng Wu Xue Bao. 2007 Jun;40(3):217-23.
In this paper, Shannong 87074-519, a derivative of wheat-decaploid Elytrigia elongata, was identified by inoculation assessment, cytological analysis, simple sequence repeat (SSR), molecular marker technique,and genomic in situ hybridization (GISH). The results are as follows: the chromosome number of Shannong87074-519 in root tip cells was 2n=44, 22 bivalents were observed in most PMC at MI, and the average chromosome configuration was 2n=44=21.82 II+0.36 I , and the chromosome configuration (2n=43=21 II +1 I) was observed in most PMC of F1 between Shannong87074-519 and C.S. at MI. Therefore, it was an alien disomic addition line with one pair chromosome of Elytrigia elongata. Then the St total genomic DNA was labeled as probe in GISH, the green-yellow hybridization signal was observed in two intact chromosomes, indicating that Shannong87074-519 was added by one pair chromosome of St genome. The SSR-PCR technique was employed in the primer filtration, and the molecular marker BARC165 was singled out from 170 primers, which could amplify the specific molecular marker BARC165(268) of Elytrigia elongata in Shannong87074-519. Subsequently, the specific segment in Elytrigia elongata was cloned and labeled as probe in GISH of root tip cells of Shannong87074-519, the light yellow hybridization signal was observed in both chromatin at interphase and chromosome at mitotic metaphase, thus the BARC165(268) could be applied as a specific molecular marker to detect alien chromatin of Elytrigia elongata in Shannong87074-519. Because of the good agronomic characteristics, high immunity to yellow rust, and dominant new yellow rust resistant gene located at the added chromosome St, assigned as YrSt temporarily, Shannong87074-519 has very important value in wheat breeding and genetics improvement.
本文通过接种鉴定、细胞学分析、简单序列重复(SSR)分子标记技术和基因组原位杂交(GISH)对小麦-十倍体长穗偃麦草衍生系山农87074-519进行了鉴定。结果如下:山农87074-519根尖细胞染色体数为2n=44,减数分裂中期Ⅰ多数花粉母细胞观察到22个二价体,平均染色体构型为2n=44=21.82Ⅱ+0.36Ⅰ,山农87074-519与中国春杂交F1的多数花粉母细胞减数分裂中期Ⅰ观察到染色体构型为(2n=43=21Ⅱ+1Ⅰ)。因此,它是一个附加一对长穗偃麦草染色体的异源二体附加系。然后以St基因组总DNA为探针进行GISH分析,在两条完整染色体上观察到绿黄色杂交信号,表明山农87074-519附加了一对St基因组染色体。利用SSR-PCR技术进行引物筛选,从170对引物中筛选出分子标记BARC165,该引物能在山农87074-519中扩增出长穗偃麦草的特异分子标记BARC165(268)。随后,克隆长穗偃麦草中的特异片段并作为探针用于山农87074-519根尖细胞GISH分析,在间期染色质和有丝分裂中期染色体上均观察到浅黄色杂交信号,因此BARC165(268)可作为检测山农87074-519中长穗偃麦草外源染色质的特异分子标记。山农87074-519农艺性状优良,对条锈病免疫,且附加的St染色体上携带显性抗条锈病新基因,暂命名为YrSt,在小麦育种和遗传改良中具有重要价值。