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载有抗逆性基因的多枝赖草染色体连锁群的特定TAC克隆的鉴定

DOI: 10.7668/hbnxb.2008.05.002, PP. 6-12

Keywords: 多枝赖草,TAC克隆,二体(易位)附加系,抗逆性,双色荧光原位杂交

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Abstract:

分别以耐黄矮病耐蚜虫小赖麦附加系Line24、耐盐耐旱小赖麦附加系Line15、耐盐小赖麦易位附加系Line14为材料,以它们各自所附加的含有相应抗逆基因的多枝赖草染色体(简称为抗逆染色体)的6个特定SSR分子标记为探针,通过两轮菌落杂交法筛选多枝赖草基因组可转化人工染色体(TAC)文库,最终得到经过验证的阳性克隆个,它们分别对应于3个(易位)附加系中所附加抗逆染色体(臂)连锁群的6个特定SSR标记,每一标记所对应的TAC克隆数目变化在5~19个,平均约13个。进一步从来自Line24的抗逆染色体连锁群的同一个探针筛选到的阳性克隆中抽查2个TAC克隆15J18和09I02,通过双色荧光原位杂交技术(Double-colorFISH)进行初步定位,直观地证明了文库筛选结果的准确性。为各条抗逆染色体的识别鉴定和相关抗逆基因的精细定位奠定了基础。

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