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铁结合蛋白(Ferritin,Fer)参与干旱胁迫应答反应,开发Fer基因抗旱相关的分子标记可为抗旱小麦品种选育提供依据。本研究依据2个强抗旱性和2个弱抗旱性小麦品种1A染色体上铁结合蛋白基因(TaFer-A1)序列的差异,开发TaFer-A1基因抗旱相关的分子标记,用150份萌发期抗旱性不同的小麦品种(系)对标记的有效性进行验证。克隆序列比对发现,TaFer-A1基因在4份抗旱性极端品种间仅存在7个变异位点,包括6个SNP位点和1个3bp碱基Del/Ins位点,其中仅在基因第1个内含子区域的3个连续碱基TCT的Del/Ins位点变异与4份品种抗旱性的表型相对应,而其余6个SNP位点在强与弱抗旱性品种之间随机发生。根据两组抗旱性极端品种间TCT碱基的差异,设计1对引物开发出了分子标记FerA1i-ntr1。该分子标记在2个强抗旱性品种中扩增出了167bp特异性条带,定名为TaFer-A1a等位变异;在2个弱抗旱性品种中扩增出了170bp特异性条带,定名为TaFer-A1b等位变异,表明FerA1i-ntr1为共显性标记。在分子标记FerA1i-ntr1检测的150份小麦品种(系)中,有73份被检测为TaFer-A1a等位变异类型,平均相对发芽率为70.1%;77份被检测为TaFer-A1b等位变异类型,平均相对发芽率为55.1%。TaFer-A1a等位变异类型品种(系)的平均相对发芽率极显著高于TaFer-A1b等位变异类型的(P<0.01),说明该共显性标记FerA1-intr1可用于小麦抗旱性的鉴定和筛选,也表明TaFer-A1基因与小麦抗旱性有关。
Iron-binding protein (Ferritin, Fer) participates in drought stress response and development of molecular markers related to drought resistance of Fer gene may provide the basis for drought-resistant wheat breeding. In this study, TaFer-A1 gene was developed based on the difference of the TaFer-A1 sequence of 1A chromosomal DNA of two wheat cultivars with strong drought resistance and two weakly drought-tolerant cultivars. Different wheat varieties (lines) validate the validity of the marker. Cloning sequence alignment revealed that TaFer-A1 had only seven polymorphic sites among the four extreme drought resistance cultivars, including six SNP sites and one 3bp Del / Ins site, of which only the first The variation of Del / Ins locus of three consecutive base TCTs in intron regions corresponded with the phenotypes of drought resistance of four cultivars, while the other six SNP loci were randomly generated between strong and weak drought tolerant cultivars. According to the difference of TCT bases between two extreme drought tolerance cultivars, a pair of primers was designed to develop a molecular marker FerA1i-ntr1. The molecular marker amplified a 167bp-specific band in two strong drought-resistance cultivars and named TaFer-A1a allelic variation. A 170bp-specific band was amplified from two weakly drought-tolerant cultivars and named TaFer-A1b allelic variation, indicating that FerA1i-ntr1 is a codominant marker. Among 150 wheat cultivars (lines) with molecular markers FerA1i-ntr1, 73 were detected as TaFer-A1a alleles with an average relative germination rate of 70.1%; 77 were detected as TaFer-A1b alleles Type, the average relative germination rate was 55.1%. The average relative germination rate of TaFer-A1a alleles was significantly higher than that of TaFer-A1b alleles (P <0.01), indicating that the codominant marker FerA1-intr1 could be used to identify the drought resistance of wheat And screening, also showed TaFer-A1 gene and wheat drought resistance.