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利用小麦品种烟农15与中间偃麦草直接杂交,以烟农15为回交亲本对其杂种F1回交2次,再经自交3次,从BC2F4中选出种质系山农0095。利用形态学、细胞学、种子醇溶蛋白酸性聚丙烯酰胺凝胶电泳(A-PAGE)、RAPD和GISH等方法对其进行鉴定。结果表明,山农0095平均株高78cm,穗长17.3cm,穗粒数74个,旗叶长36.3cm,旗叶宽3.03cm,茎秆粗壮,多花多实,繁茂性好;其根尖细胞染色体数目为2n=42,花粉母细胞减数分裂中期Ⅰ(PMCMⅠ)染色体构型为2n=21Ⅱ;它与烟农15的杂种F1在减数分裂中期Ⅰ的大多数花粉母细胞能观察到2个单价体,平均染色体构型为2n=20.08Ⅱ+1.84Ⅰ。A-PAGE鉴定结果表明,山农0095在β区出现一条中间偃麦草的特异带;从180个RAPD引物中筛选出一个特异引物S186(5′-GATACCTCGG-3′),能够在山农0095中扩增出一条约900bp中间偃麦草的特异带,标记为S186900;以中间偃麦草基因组DNA为标记探针、中国春基因组DNA为封阻的原位杂交结果进一步表明,山农0095的42条染色体中含有2条完整的中间偃麦草染色体,是小麦-中间偃麦草的二体异代换系。
Wheat germplasm Yannong 15 and Thinopyrum intermedium were directly hybridized, and their hybrid F1 was backcrossed twice with backcrossing parents of tobacco cultivar Nongnong 15, followed by selfing for 3 times to select germplasm Shannong 0095 from BC2F4. Morphological, cytological, seed gliadin acidic polyacrylamide gel electrophoresis (A-PAGE), RAPD and GISH and other methods to identify them. The results showed that Shannong 0095 had an average plant height of 78cm, a spike length of 17.3cm, a number of spikelets of 74, a flag leaf length of 36.3cm and a flag leaf width of 3.03cm. The stalk was sturdy, The number of chromosomes was 2n = 42, and the chromosome configuration of PMCMⅠ was 2n = 21Ⅱ. The hybrid F1 with tobacco cultivar Nongnong 15 could be observed in most pollen mother cells of metaphase Ⅰ A unit price, the average chromosome configuration of 2n = 20.08 Ⅱ + 1.84 Ⅰ. The results of A-PAGE showed that Shannong 0095 appeared a specific band in the β region and a specific primer S186 (5’-GATACCTCGG-3 ’) was screened out from 180 RAPD primers, which can be used in Shannon 0095 A specific band about 900bp of Agropyron chinense was amplified and labeled as S186900. The results of in situ hybridization with Chinese spring genomic DNA as a marker probe using the genome DNA of T. triticina further showed that 42 chromosomes of Shannong 0095 Contains two complete chromosomes of Th. Intermedium, and is a two-body alien substitution line of wheat-Thinopyrum intermedium.