CAO Yang,LUO Shunzhen,HE Chunmei,et al.Mating-type Molecular Identification of Phlebopus portentosus Homokaryotic Strains[J].Northern Horticulture,2024,(22):101-108.[doi:10.11937/bfyy.20241213]
暗褐脉柄牛肝菌同核菌株交配型的分子鉴定
- Title:
- Mating-type Molecular Identification of Phlebopus portentosus Homokaryotic Strains
- 文章编号:
- 1001-0009(2024)22-0101-08
- Keywords:
- Phlebopus portentosus; mating-type; identification; SNP marker
- 分类号:
- S 646.3
- 文献标志码:
- A
- 摘要:
- 以暗褐脉柄牛肝菌异核体及其担孢子萌发所得的同核菌株为试材，采用基于β-flanking基因和ste3.2基因开发分子标记的方法，研究了暗褐脉柄牛肝菌同核菌株分子鉴定的手段，以期为提高其杂交育种的工作效率提供参考依据。结果表明:用于扩增β-flanking基因区域的5对引物，产物叠加覆盖该基因91%的区域。用于扩增ste3.2基因区域的5对引物，产物叠加覆盖该基因71%的区域。10对引物在2个异核亲本中均能扩出，但可用序列长度及SNP位点数目有差异。根据第一轮PCR扩增结果，选择beta2和3.22引物对HZ19110的同核菌株进行第二轮PCR扩增并分型，交配型为A1B1、A2B2、A1B2、A2B1的同核菌株分别有5、4、4、4株。选择beta5和3.25引物对HZ20029的同核菌株进行第二轮PCR扩增并分型，交配型为A3B3、A4B4、A3B4、A3B4的同核菌株分别有4、3、3、3株。上述分型结果与两两对峙培养、镜检锁状联合验证交配型结果一致。该研究中开发的分子鉴定方法能够快速、准确地对来源不同异核亲本的暗褐脉柄牛肝菌同核菌株进行交配型鉴定，具有一定的通用性，可有效提升杂交育种工作效率。
- Abstract:
- Heterokaryotic parent strains and their offspring homokaryotic strains of Phlebopus portentosus were used as the test materials.β-flanking gene and ste3.2 gene，which respectively represent mating-type A and B genotype，was used to develop molecular markers to study the mating-type identification of P.portentosus homokaryotic strains,in order to provide reference for improving the work efficiency of its cross breeding.The results showed that 5 pairs the primers could amplify 91% of the β-flanking gene region，other 5 pairs could amplify 71% of the ste3.2 gene region.Amplification sequencing was successfully performed on 2 heterokaryotic parent strains using all primers above although the usable sequence lengths and SNP sites numbers were visibly different.Based on the results of first round PCR，primers beta2 and 3.22 were chosen for the second round PCR and mating-typing of HZ19110 homokaryotic strains.There were 5，4，4 and 4 homokaryotic strains of mating-type A1B1,A2B2，A1B2，and A2B1 respectively.Primers beta5 and 3.25 were chosen for the second round PCR and mating-typing of HZ20029 homokaryotic strains.There were 4，3，3 and 3 homokaryotic strains of mating-type A3B3,A4B4，A3B4，and A4B3 respectively.The mating-type identification of homokaryotic strains was consistent with the results of pairwise culture and clamp screening under microscope.Mating-type of P.portentosus homokaryotic strains from different heterokaryotic parent strains could be rapidly and accurately identified by using the molecular methods in this study.The breeding efficiency of this species could be consequently improved.
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备注/Memo
第一作者简介：曹旸（1983-），女，博士，副研究员，现主要从事食用菌学等研究工作。E-mail：caoyang8352@163.com.责任作者：纪开萍（1966-），女，本科，研究员，现主要从事牛肝菌人工驯化及栽培等研究工作。E-mail：jkpcnlc@126.com.基金项目：云南省科技计划资助项（202104AP080138）；黔西南州科技计划资助项目（2022-1-48）；云南省市场监督管理局科技计划资助项目（2022YSJK01）。收稿日期：2024-03-25