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中国板栗与美洲栗杂种抗性遗传标记——莽草酸脱氢酶(SKD)的遗传分析

Genetic Analysis of Shikimate Dehydrogenase Allozymes,a Genetic Marker Resisting to Chustnut-blight,in Hybrid of Castanea mollissima×C.dentata

  • 摘要: 采用超薄平板聚丙烯酰胺等电聚焦技术和单株家系、杂交家系分析方法对中国板栗(Castanea mollissima)和美洲栗(C.dentata)莽草酸途径的关键性酶——莽草酸脱氢酶(SKD)的遗传变异进行了研究。对单株家系、种内杂交家系和种间回交BC1群体的研究结果表明:栗属莽草酸脱氢酶共有4个活性区域,活性基因座具有物种特异性,并符合孟德尔式遗传,美洲栗的Skd-1与Skd-2为独立分离基因座,但中国板栗的Skd-3与Skd-4则为紧密连锁基因座。通过对中国板栗和美洲栗杂种F2代群体分离的杂种酶谱表型分析,表明中国板栗Skd-3,-4连锁群可能在基因组中存在重复,并推断美洲栗的Skd-1和Skd-2可能分别由重复连锁群独自变异演替而成。莽草酸脱氢酶重复基因座的存在并表达出酶活性,可能使中国板栗的莽草酸途径更有效率,增强中国板栗在受到栗疫病菌等微生物攻击时的反应应答,从而提高其抗性。研究认为莽草酸脱氢酶有作为抗性辅助选择遗传标记的潜力,在回交育种拯救美洲栗计划中具有潜在的应用价值。

     

    Abstract: In plants,the shikimate pathway is an important metabolism pathway in response to wounding and fungal attack.The genetic analysis of enzymes in the pathway can provide the basic information about their applicability as a genetic marker.In this study,variability and inheritance of shikimate dehydrogenase (SKD;EC 1.1.1.25 ) allozyme,one of the key enzymes in shikimate pathway,was investigated in Castanea mollissima and C.dentata using isoelectric focusing in thin-layer polyacrylamide slad gels.The single-tree progeny method and intra-and interspecific controlled cross families were used for the analyses.The results showed SKD had four activity zones and exhibited species specific banding patterns in Castanea.Two alleles were detected at each four locus and inherited in Mendelian fashion.Locus Skd-1 and Skd-2 unique to C.dentata seemed to be segregated independantly,while a highly significant linkage was observed between two loci Skd-3 and Skd-4 unique to C.mollissima.However,the phenotypic analysis of SKD allozyme banding patterns in F2 populations of C.mollissima×C.dentate revealed that the linkage group between Skd-3 and Skd-4 maybe have been replicated in C.mollissima.It is speculated that Skd-1 and Skd-2 in C.dentata possibly come from repetition linkage groups during a speciation process.Because of expression of the double SKD loci activity,the shikimate pathway in C.mollissima is considered more efficient and consequently,responses of C.mollissima to Cryphonectria parasitica invasion and other microbial attack may be elevated.So the resistance of C.mollissima to chestnut blight is considered increase.This study suggested that SKD allozyme could be a potential genetic marker for aided selection on blight-resistance and have potential application in the backcross-breeding program of C.dentata.

     

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