基于MPMS方法的黑木耳优良菌株选育

doi:10.3969/j.issn.1000-1565.2022.06.013

摘要/Abstract

摘要： 为获得高产优质的黑木耳菌株,以黑木耳(Auricularia auricula)菌株Aa66为出发菌株,采用多功能等离子体诱变系统(multifunction plasma mutagenesis system,MPMS)进行诱变育种实验,共获得315株诱变菌株.通过初筛拮抗实验及复筛对菌株长速长势、酶活及生物学效率的测定,共得到8株优良诱变菌株,且使用简单重复序列区间扩增多态性分子标记技术(inter-simple sequence repeat,ISSR)在分子水平上确定其均与出发菌株Aa66有明显差异,其中优良诱变菌株Y9生长速度最快,为4.98 mm/d,较出发菌株Aa66提高0.54 mm/d,生物学效率为115.34%,较出发菌株Aa66提高8.43%,表明MPMS可以作为一种选育优良黑木耳菌株的新方法.

关键词: 黑木耳, 多功能等离子体诱变系统(MPMS), 诱变, 分子标记

Abstract: In order to obtain high-yield and high-quality Auricularia auricula strains, using A. auricula strain Aa66 as the starting strain, multifunction plasma mutagenesis system(MPMS)was used to carry out mulation breeding experiments, and a total of 315 mutant strains were obtained. The antagonism experiment and re-screening were used to determine the growth rate, enzyme activity and biological efficiency of the strain. Through the preliminary screening antagonistic experiment and the determination of strain growth rate,growth trend,enzyme activity and biological efficiency by re-screening, a total of 8 excellent mutagenic strains were obtained, and the inter-simple sequence repeat(ISSR)technology was used to amplify polymorphic molecular markers. They were determined to be significantly different from the starting strain Aa66 at the molecular level. Among them, the excellent mutant strain Y9 grew with the fastest speed of 4.98 mm/d, which was 0.54 mm/d higher than that of the starting strain Aa66, and the biological efficiency was 115.34%, which was 8.43% higher than that of the starting strain Aa66, indicating that MPMS can be used as a new method for breeding excellent A. auricula strains. - DOI:10.3969/j.issn.1000-1565.2022.06.013基于MPMS方法的黑木耳优良菌株选育尹显达¹,刘敏¹,杨超上¹,张铁军²,王谦¹(1.河北大学生物工程技术创新中心,河北保定 071002;2.承德市农业环境保护监测站,河北承德 067000)摘要:为获得高产优质的黑木耳菌株,以黑木耳(Auricularia auricula)菌株Aa66为出发菌株,采用多功能等离子体诱变系统(multifunction plasma mutagenesis system,MPMS)进行诱变育种实验,共获得315株诱变菌株.通过初筛拮抗实验及复筛对菌株长速长势、酶活及生物学效率的测定,共得到8株优良诱变菌株,且使用简单重复序列区间扩增多态性分子标记技术(inter-simple sequence repeat,ISSR)在分子水平上确定其均与出发菌株Aa66有明显差异,其中优良诱变菌株Y9生长速度最快,为4.98 mm/d,较出发菌株Aa66提高0.54 mm/d,生物学效率为115.34%,较出发菌株Aa66提高8.43%,表明MPMS可以作为一种选育优良黑木耳菌株的新方法.关键词:黑木耳;多功能等离子体诱变系统(MPMS);诱变;分子标记中图分类号:S646.6 文献标志码:A 文章编号:1000-1565(2022)06-0650-07Breeding of fine strains of Auricularia auriculata based on MPMS methodYIN Xianda¹, LIU Min¹, YANG Chaoshang¹, ZHANG Tiejun², WANG Qian¹(1.Bioengineering Technology Innovation Center, Hebei University, Baoding 071002, China; 2.Chengde City Agricultural Environmental Protection Monitoring Station, Chengde 067000, China)Abstract: In order to obtain high-yield and high-quality Auricularia auricula strains, using A. auricula strain Aa66 as the starting strain, multifunction plasma mutagenesis system(MPMS)was used to carry out mulation breeding experiments, and a total of 315 mutant strains were obtained. The antagonism experiment and re-screening were used to determine the growth rate, enzyme activity and biological efficiency of the strain. Through the preliminary screening antagonistic experiment and the determination of strain growth rate,growth trend,enzyme activity and biological efficiency by re-screening, a total of 8 excellent mutagenic strains were obtained, and the inter-simple sequence repeat(ISSR)technology was used to amplify polymorphic molecular markers. They were determined to be significantly different from the starting strain Aa66 at the molecular level. Among them, the excellent mutant strain Y9 grew with the fastest speed of 4.98 mm/d, which was 0.54 mm/d higher than that of the starting strain Aa66, and the biological efficiency was 115.34%, which was 8.43% higher than that of the starting strain Aa66, indicating that MPMS can be used as a new method for breeding excellent A. auricula strains. - 收稿日期:2022-04-28 基金项目:河北省现代农业食用菌产业技术体系创新团队(第2期)(HBCT2018050204) 第一作者:尹显达(1996—),男,河北廊坊人,河北大学在读硕士研究生,主要从事大型真菌研究.E-mail:yin19965@sina.com 通信作者:王谦(1962—),男,北京人,河北大学研究员,主要从事食药用真菌研究与开发工作.E-mail:wq6203_cn@126.com第6期尹显达等:基于MPMS方法的黑木耳优良菌株选育

Key words: Auricularia auricula, multifunction plasma mutagenesis system(MPMS), mutation, molecular markers

中图分类号:

S646.6

尹显达,刘敏,杨超上,张铁军,王谦. 基于MPMS方法的黑木耳优良菌株选育[J]. 河北大学学报(自然科学版), 2022, 42(6): 650-656.

YIN Xianda, LIU Min, YANG Chaoshang, ZHANG Tiejun, WANG Qian. Breeding of fine strains of Auricularia auriculata based on MPMS method[J]. Journal of Hebei University(Natural Science Edition), 2022, 42(6): 650-656.

参考文献

[1] 吴芳,员瑗,刘鸿高,等.木耳属研究进展[J].菌物学报, 2014, 33(2): 198-207. DOI:10.13346/j.mycosystema.130282.
[2] 戴玉成,杨祝良.中国药用真菌名录及部分名称的修订[J].菌物学报, 2008, 27(6): 801-824. DOI:10.3969/j.issn.1672-6472.2008.06.001.
[3] 刘文贺,苏玲,王琦.不同产区黑木耳中营养成分比较分析[J].北方园艺, 2020(5): 121-128.
[4] 罗晓莉,张沙沙,曹晶晶,等.云南3种胶质食用菌营养成分分析与蛋白质营养价值评价[J].食品工业科技, 2021, 42(14): 328-333. DOI:10.13386/j.issn1002-0306.2020090143.
[5] LU A X, YU M G, SHEN M, et al. Preparation of the Auricularia auricular polysaccharides simulated hydrolysates and their hypoglycaemic effect[J]. Int J Biol Macromol, 2018, 106: 1139-1145. DOI:10.1016/j.ijbiomac.2017.08.118.
[6] SUN S J, ZHANG X J, SUN S W, et al. Production of natural melanin by Auricularia auricula and study on its molecular structure[J]. Food Chem, 2016, 190: 801-807. DOI:10.1016/j.foodchem.2015.06.042.
[7] BOONSONG S, KLAYPRADIT W, WILAIPUN P. Antioxidant activities of extracts from five edible mushrooms using different extractants[J]. Agric Nat Resour, 2016, 50(2): 89-97. DOI:10.1016/j.anres.2015.07.002.
[8] ZHANG J, SUN T T, WANG S X, et al. Transcriptome exploration to provide a resource for the study of Auricularia heimuer[J]. J For Res, 2020, 31(5): 1881-1887. DOI:10.1007/s11676-019-00989-8.
[9] 李江宇,迟蔚,刘斌,等.我国黑木耳产业经济发展现状及生产效益对比分析[J]. 食用菌, 2019, 41(6): 6-8.
[10] 寇博翔,杨嘉雪,杨林英,等.利用多功能等离子体诱变技术选育糙皮侧耳新菌株[J].食用菌学报, 2021, 28(4): 7-14. DOI:10.16488/j.cnki.1005-9873.2021.04.002.
[11] 王晨,马欣欣,平琳琳,等.ARTP诱变技术选育黑木耳优良发酵菌株初探[J].中国食用菌, 2021, 40(3): 17-22. DOI:10.13629/j.cnki.53-1054.2021.03.004.
[12] 吴亚楠,尹显达,耿培妍,等.基于MPMS诱变体系的茶树菇细胞工程育种[J].中国食用菌, 2021, 40(8): 18-23. DOI:10.13629/j.cnki.53-1054.2021.08.005.
[13] 中华人民共和国农业部.食用菌菌种区别性鉴定拮抗反应:NY/T 1845—2010[S].北京:中国农业出版社, 2010.
[14] 李碧琼,陈政明,林俊扬,等.茶树菇高效栽培优良菌株筛选试验研究[J].中国食用菌, 2012, 31(5): 6-8. DOI:10.13629/j.cnki.53-1054.2012.05.013.
[15] 回晶,赵文静,邹志远,等.金耳液体培养过程中几种胞外酶活性的变化规律[J].食用菌学报, 2007, 14(3): 29-32. DOI:10.16488/j.cnki.1005-9873.2007.03.003.
[16] 李守勉,李明,田景花,等.八个杏鲍菇菌株胞外酶活性及蛋白质含量的研究比较[J].食用菌, 2007, 29(6): 11-12.
[17] 宋爱荣,郭立忠,刘作亭,等.七个白色金针菇菌株发酵液中四种胞外酶活性的测定与分析[J].中国食用菌, 1999, 18(4):31-34.
[18] 韩增华,张丕奇,孔祥辉,等.黑木耳胞外酶活变化与栽培性状比较的研究[J].食用菌学报, 2007, 14(4): 41-46. DOI:10.16488/j.cnki.1005-9873.2007.04.006.
[19] 中华人民共和国农业部.食用菌菌种真实性鉴定 ISSR法:NY/T 1730—2009[S].北京:中国标准出版社, 2009.
[20] JEDRZEJCZYK I, REWERS M. Genome size and ISSR markers for Mentha L.(Lamiaceae)genetic diversity assessment and species identification[J]. Ind Crops Prod, 2018, 120: 171-179. DOI:10.1016/j.indcrop.2018.04.062.
[21] HAIDER N. Determining phylogenetic relationships among date palm cultivars using random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR)markers[J]. Methods Mol Biol, 2017, 1638: 153-172. DOI:10.1007/978-1-4939-7159-6_14. (