应用原生质体融合技术培育平菇与杏鲍菇优良新菌株

doi:10.3969/j.issn.1004-1524.2020.08.09

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (8): 1397-1404.DOI: 10.3969/j.issn.1004-1524.2020.08.09

应用原生质体融合技术培育平菇与杏鲍菇优良新菌株

武秋颖¹, 张运峰¹, 张淑红¹, 高凤菊¹, 刘海英², 范永山^1,*

1.唐山师范学院生命科学系,河北唐山063000;
2.唐山市农业科学研究院蔬菜研究所,河北唐山 063000

收稿日期:2020-01-16 出版日期:2020-08-25 发布日期:2020-08-28
通讯作者: *,范永山,E-mail: fanyongshan@126.com
作者简介:武秋颖(1985—),女,河北唐山人,硕士,实验师,研究方向为生物化学与微生物学。E-mail:wuqiuying123@126.com
基金资助:
唐山市科技计划(17120204a); 唐山市人才资助项目(A201903018); 唐山师范学院科学基金(2017C03,2016C05)

Application of protoplast fusion technology to breed new strains of Pleurotus ostreatus and Pleurotus eryngii

WU Qiuying¹, ZHANG Yunfeng¹, ZHANG Shuhong¹, GAO Fengju¹, LIU Haiying², FAN Yongshan^1,*

1. Department of Life Sciences, Tangshan Normal College, Tangshan 063000, China;
2. Institute of Vegetables,Tangshan Academy of Agricultural Sciences, Tangshan 063000, China

Received:2020-01-16 Online:2020-08-25 Published:2020-08-28

摘要/Abstract

摘要： 为了培育出具有优良互补性状的平菇和杏鲍菇新菌株,以高产、抗杂能力强的平菇品种CCEF89和优质、适应性强的杏鲍菇品种PL7为亲本菌株,建立原生质体聚乙二醇(PEG)融合体系,得到最佳融合条件:两亲本原生质体数量按1∶1混合,30% PEG 6000,0.01 mol·L^-1Ca²⁺,32 ℃水浴30 min,融合率达0.010 1%~0.028 2%。通过拮抗反应试验和Rep-PCR分子鉴定,从515个融合再生菌株中获得12个融合菌株P1~P12;出菇试验表明,P1和P5为杏鲍菇新菌株,其他为平菇新菌株。P1和P5的产量、生物学效率均显著高于其亲本菌株PL7,并且抗杂能力显著提高;与亲本菌株CCEF89相比,平菇新菌株P4、P10和P11长满培养料和形成原基的时间缩短5.5~7.3 d,第1茬平均产量提高18.0%~24.0%,总生物学效率提高21.00%~27.67%。利用原生质体融合技术培育出了具有亲本优良互补性状的平菇和杏鲍菇新菌株,为同时进行2种不同食用菌的种质资源创新和新品种选育提供了一种新途径。

关键词: 原生质体融合, 平菇, 杏鲍菇, 融合菌株, 种质资源

Abstract: In order to breed new strains combining the elite complementary traits of Pleurotus ostreatus and Pleurotus eryngii, Pleurotus ostreatus variety CCEF89 with the properties of high yield and strong ability to resist contamination, and Pleurotus eryngii variety PL7 with the properties of high quality and strong adaptability were used as the parent strains to build the PEG (polyethylene glycol)protoplast fusion system.Under the optimal protoplast fusion condition of 1∶1 quantity ratio of the parent strains, 32 ℃ water bath, 30% PEG 6000, 0.01 mol·L^-1 Ca²⁺ and 30 min co-incubation, the fusion rate was up to 0.010 1%-0.028 2%. 12 protoplast fusants (P1-P12) were identified by antagonistic reaction test and Rep-PCR molecular identification from 515 protoplast fusion regeneration isolates. After the mushroom cultivation experiments, P1 and P5 were identified as new strains of P. eryngii, and the other 10 protoplast fusants were new strains of P. ostreatus. The mushroom yield and biological efficiency of new strains P1 and P5 were significantly higher than those of their parent PL7, and their ability to resist contamination were increased significantly. Comparing to the parent CCEF89, the time of medium cultivation and mushroom primordia formation of P. ostreatus new strains P4, P10 and P11 were 5.5-7.3 days earlier, the yield of the first harvest mushroom and the total biological efficiency increased by 18.0%-24.0% and 21.00%-27.67%, respectively.Using the protoplast fusion technology, we acquired P. ostreatus and P. eryngii new strains with excellent complementary properties of the parents simultaneously, and proposed a new pathway to innovate new germplasm resources and bred new varieties of two different edible fungiat the same time.

Key words: protoplast fusion, Pleurotus ostreatus, Pleurotus eryngii, fusant, germplasm resource

中图分类号:

S646.1⁺4

武秋颖, 张运峰, 张淑红, 高凤菊, 刘海英, 范永山. 应用原生质体融合技术培育平菇与杏鲍菇优良新菌株[J]. 浙江农业学报, 2020, 32(8): 1397-1404.

WU Qiuying, ZHANG Yunfeng, ZHANG Shuhong, GAO Fengju, LIU Haiying, FAN Yongshan. Application of protoplast fusion technology to breed new strains of Pleurotus ostreatus and Pleurotus eryngii[J]. , 2020, 32(8): 1397-1404.

参考文献

[1] 桂明英, 王刚, 郭永红, 等. 食用菌育种技术的研究进展[J]. 中国食用菌, 2006, 25(5): 3-5.
GUI M Y, WANG G, GUO Y H, et al. Research aadvancement of breeding technique about edible fungi[J]. Edible Fungi of China, 2006, 25(5): 3-5.(in Chinesewith English abstract)
[2] 付立忠, 吴学谦, 魏海龙, 等. 我国食用菌育种技术应用研究现状与展望[J]. 食用菌学报, 2005, 12(3): 63-68.
FU L Z, WU X Q, WEI H L, et al. Advances in the study on breeding technique of edible fungi in China and the prospect[J]. Acta Edulis Fungi, 2005, 12(3): 63-68.(in Chinesewith English abstract)
[3] 贺立虎, 李黔蜀. 食用菌遗传育种技术的研究进展[J]. 陕西农业科学, 2012, 58(1): 116-118.
HE L H, LI Q S. Advances on genetic breeding technologies of edible fungi[J].Shaanxi Journal of Agricultural Sciences, 2012, 58(1): 116-118.(in Chinese)
[4] 康林芝, 熊荣园, 金磊, 等. 金针菇与平菇远缘亲本原生质体融合研究[J]. 食品研究与开发, 2019, 40(19): 210-214.
KANG L Z, XIONG R Y, JIN L, et al. The research of protoplast fusion of Flammulina velutipes and Pleurotus ostreatus[J]. Food Research and Development, 2019, 40(19): 210-214.(in Chinese with English abstract)
[5] 王珂. 羊肚菌菌核发育形态、IMV检测及原生质体融合技术研究[D]. 郑州: 郑州轻工业大学,2019.
WANG K. Study on morphology of sclerotial development, IMV detection and protoplast fusion of Morchella mushrooms [D].Zhengzhou: Zhengzhou University of Light Industry, 2019. (in Chinese with English abstract)
[6] 杨珊. 猴头菌单核体杂交及原生质体融合品质育种的研究[D]. 上海: 上海海洋大学, 2018.
YANG S. Study on mononuclear hybridization and protoplast fusion quality breeding of Hericium erinaceus[D]. Shanghai: Shanghai Ocean University, 2018.(in Chinese with English abstract)
[7] 张运峰, 张淑红, 武秋颖, 等. 平菇原生质体再生菌株的ISSR-PCR遗传多样性分析[J]. 河北农业大学学报, 2016, 39(2): 100-104.
ZHANG Y F, ZHANG S H, WU Q Y, et al. Genetic diversity analysis of protoplast regeneration isolates of Pleurotus ostreatus by ISSR-PCR markers[J]. Journal of Agricultural University of Hebei, 2016, 39(2): 100-104.(in Chinese with English abstract)
[8] 董汪洋, 王郑隆, 杨云乔, 等. 杏鲍菇的菌株筛选与原生质体制备条件优化[J]. 浙江农业学报, 2015, 27(5): 782-786.
DONG W Y, WANG Z L, YANG Y Q, et al. Screening and optimization on protoplast preparation conditions of Pleurotus eryngii[J]. Acta Agriculturae Zhejiangensis, 2015, 27(5): 782-786.(in Chinese with English abstract)
[9] 张国锁, 张淑红, 张运峰, 等. 杏鲍菇多糖对糖尿病小鼠的降血糖作用[J]. 中国食用菌, 2019, 38(1): 38-40.
ZHANG G S, ZHANG S H, ZHANG Y F, et al. Hypoglycemic effect of Pleurotus eryngii polysaccharides on diabetic mice[J]. Edible Fungi of China, 2019, 38(1): 38-40.(in Chinese with English abstract)
[10] 李小六, 张运峰, 张淑红, 等. 原生质体EMS诱变选育杏鲍菇高产菌丝多糖菌株[J]. 核农学报, 2017, 31(8): 1463-1468.
LI X L, ZHANG Y F, ZHANG S H, et al. Selection of Pleurotus eryngii strains with high yield of mycelial polysaccharide by EMS protoplast mutagenesis[J]. Journal of Nuclear Agricultural Sciences, 2017, 31(8): 1463-1468.(in Chinese with English abstract)
[11] 刘海英, 张运峰, 范永山, 等. 紫外线对杏鲍菇原生质体的诱变作用[J]. 核农学报, 2011, 25(4): 719-723.
LIU H Y, ZHANG Y F, FAN Y S, et al. UV-mutagenesis of protoplasts of Pleurotus eryngii[J]. Journal of Nuclear Agricultural Sciences, 2011, 25(4): 719-723.(in Chinese with English abstract)
[12] 虞磊,李蕤.食用菌原生质体融合育种技术及其进展[J].合肥联合大学学报, 2000, 10(4): 97-100.
YU L, LI R. Review on the fusion breeding technology of protoplast from domestic fungus[J]. Journal of Hefei Union University, 2000, 10(4): 97-100. (in Chinese with English abstract)
[13] 孙娇娇. 松乳菇与平菇原生质体融合的初步研究[D]. 南昌: 江西农业大学, 2012.
SUN J J. Preliminary study on protoplast fusion of Lactarius delicious and Pleurotus ostreatus[D]. Nanchang: Jiangxi Agricultural University, 2012.(in Chinese with English abstract)
[14] 杨土凤, 王立洪, 杨涛, 等. 平菇与香菇原生质体融合新菌株的生物学特性研究[J]. 四川大学学报(自然科学版), 2010, 47(1): 202-206.
YANG T F, WANG L H, YANG T, et al. Studies on the biological properties of novel strains from protoplast fusants between Pleurotus ostreatus and Lentinus edodes[J]. Journal of Sichuan University (Natural Science Edition), 2010, 47(1): 202-206.(in Chinese with English abstract)
[15] FUKUDA M, WAKAYAMA M, UCHIDA M, et al. Introduction of mitochondrial DNA from Pleurotus ostreatus into Pleurotus pulmonarius by interspecific protoplast fusion[J]. Journal of Wood Science, 2007, 53(4): 339-343.
[16] SELVAKUMAR P, RAJASEKAR S, BABU A G, et al. Improving biological efficiency of Pleurotus strain through protoplast fusion between P. ostreatus var. florida and P. djamor var.roseus[J]. Food Science and Biotechnology, 2015, 24(5): 1741-1748.
[17] 孙晓瑞. 秀珍菇与榆黄蘑原生质体融合育种技术研究[D]. 太古: 山西农业大学,2018.
SUN X R. Technical research of protoplast fusion between Pleurotus geesterani and Pleurotus citrinopileatus sing [D]. Taigu: Shanxi Agricultural University, 2018. (in Chinese with English abstract)
[18] 任轩, 贾乐, 宋振, 等. 双亲灭活原生质体电诱导融合法选育高漆酶活性菌株[J]. 山东农业科学, 2008, 40(8): 66-69.
REN X, JIA L, SONG Z, et al. Screening for strain with high laccase activity by protoplast electrofusion of inactivated parental strains[J]. Shandong Agricultural Sciences, 2008, 40(8): 66-69.(in Chinese with English abstract)
[19] 张甜甜,张治中,张墨楠,等.食用菌原生质体融合育种技术的研究进展[J].浙江食用菌,2009,17(3):29-31.
ZHANG T T, ZHANG Z Z, ZHANG M N, et al. Advances in the protoplast fusion technique in edible mushroom breeding[J]. Edible Fungi of Zhejiang, 2009,17(3): 29-31.(in Chinese)

应用原生质体融合技术培育平菇与杏鲍菇优良新菌株

Application of protoplast fusion technology to breed new strains of Pleurotus ostreatus and Pleurotus eryngii

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