贵州小麦品种(系)籽粒低PPO活性种质资源筛选

Screening of the low PPO activity germplasm resources in seeds of wheat variety(line)in Guizhou

【目的】筛选出小麦籽粒低多酚氧化酶(PPO)活性的种质资源,为贵州小麦籽粒品质的遗传改良及育种提供参考依据。【方法】以生产中表现优异的135份贵州小麦品种(系)为材料,采用苯酚染色法进行染色,在此基础上以STS分子标记(PPO16、PPO18和PPO29)检测小麦PPO活性相关基因,并通过Fragment AnalyzerTM毛细管电泳鉴定相关基因,进而对小麦籽粒低PPO活性基因的组成进行鉴定和筛选。【结果】135份小麦材料籽粒经苯酚染色后,有4份材料未染色(A级),9份呈浅绿色(B级),65份材料呈棕色(C级),57份材料呈黑色(D级)。STS分子标记检测结果表明,在A级和B级材料中检测到10份材料含Ppo-A1b基因、4份材料含Ppo-D1a基因,其中4份材料同时含有Ppo-A1b/Ppo-D1a基因,分别是贵麦2号、惠水1-23、石无芒和08-9选单-2-7;在C级材料中检测到3份材料含Ppo-A1b基因、17份材料含Ppo-D1a基因,其中1份材料(贵农08-9)同时含有Ppo-A1b/Ppo-D1a基因;在D级材料中均未检测到Ppo-A1b和Ppo-D1a基因。【结论】采用苯酚染色法结合STS分子标记检测可对小麦籽粒是否含低PPO活性基因进行准确鉴定。贵麦2号、惠水1-23、石无芒、08-9选单-2-7和贵农08-9等5份含有双低PPO活性基因(Ppo-A1b/Ppo-D1a)的种质资源,可作为亲本材料直接用于小麦籽粒低PPO活性遗传改良及新品种选育。

【Objective】The low PPO activity germline resources in seeds of wheat variety(line)were selected to provided a reference for genetic improvement and breeding of grain in Guizhou wheat(Triticum aestivum L.).【Method】Using 135 Guizhou wheat varieties(lines)with sound performance in planting as materials,they were stained by phenol staining method.Then the PPO activity related genes of wheat were detected with STS molecular marker(PPO16,PPO18 and PPO29),and compositions of low PPO activity related genes were identified by Fragment AnalyzerTM capillary electrophoresis.【Result】The results of phenol staining showed that among 135 wheat materials,4 materials were unstained(A level),9 materials were light green(B level),65 materials were brown(C level)and 57 materials were black(D level).The results of STS molecular marker detection showed that in A and B levels materials,10 materials contained Ppo-A1b gene,4 materials contained Ppo-D1a gene,4 materials contained Ppo-A1b/Ppo-D1a genes(Guimai 2,Huishui 1-23,Shiwumang and 08-9 Xuandan-2-7).In the C level materials,3 materials contained Ppo-A1b gene and 17 materials contained Ppo-D1a gene,1 material contained Ppo-A1b/Ppo-D1a gene at the same time.No Ppo-A1b and Ppo-D1a genes were detected in level D materials.【Conclusion】Phenol staining and STS molecular marker are combined to identify whether wheat grains contain low PPO activity genes.There are only 5 germplasm resources with low PPO activity containing Ppo-A1b/Ppo-D1a gene,which are Guimai 2,Huishui 1-23,Shiwumang,08-9 Xuandan-2-7 and Guinong 08-9,which can be applied as parents to genetic improvement and new variety breeding of wheat with low PPO activity.