平菇单核体与双核体差异的RAPD分析

张朝辉; 王勋; 陈天航; 梅会元; 韩盼盼; 刘天翔; 王振河; 邱立友

doi:10.19303/j.issn.1008-0384.2022.005.013

平菇单核体与双核体差异的RAPD分析

doi: 10.19303/j.issn.1008-0384.2022.005.013

张朝辉^{1, 2,},
王勋¹,
陈天航²,
梅会元²,
韩盼盼²,
刘天翔¹,
王振河²,
邱立友^1, ,

1.
河南农业大学生命科学学院/农业农村部农业微生物酶工程重点实验室，河南　郑州　450002
2.
河南科技学院生命科技学院，河南　新乡　453003

基金项目: 河南省科技攻关计划项目（202102110019、212102110430）；国家级大学生创新创业训练计划项目（202110467035X）

详细信息

作者简介:
张朝辉（1983−），男，博士，副教授，主要从事食药用菌生理生化研究（E-mail：p686@163.com）

通讯作者:
邱立友（1963−），男，博士，教授，主要从事真菌学研究（E-mail：qliyou@henau.edu.cn）

中图分类号: S 646.1+4
计量
- 文章访问数: 229
- HTML全文浏览量: 60
- PDF下载量: 16
- 被引次数: 0
出版历程
- 收稿日期: 2022-03-02
- 修回日期: 2022-04-04
- 刊出日期: 2022-05-28

RAPD Analysis on Differences between Monokaryotic and Dikaryotic Pleurotus ostreatus

1.
College of Life Sciences, Henan Agricultural University/Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture and Rural Affairs, Zhengzhou, Henan　450002, China
2.
College of Life Science and Technology, Henan Institute of Science and Technology, Xinxiang, Henan　453003, China

摘要

摘要: 目的探究平菇单双核菌丝体间的差异，为平菇品种培育提供技术理论基础。方法从平菇TD300的担孢子中分离得到两个单核菌株MK13和MK3，通过交配得到优良双核菌株DK300，进行平板试验和出菇试验，测定4个菌株的菌丝生长状况和出菇产量，再利用随机扩增多态性DNA（Randomly Amplified Polymorphic DNA，RAPD）标记技术检测4个菌株之间的遗传差异性。结果 DK300与亲本TD300在菌丝生长状况和产量上没有显著性差异，但MK13、MK3和交配所得的DK300与亲本TD300间存在较大的遗传差异性，而MK13、MK3和DK300间遗传差异性较小，尤其是MK3和DK300间遗传差异性最小。结论经过减数分裂形成担孢子时可能发生有基因重组，而通过两个单核菌株交配得到双核菌株过程则较少发生基因重组。
- 平菇 /
- 单核体 /
- 双核体 /
- 产量 /
- 遗传差异性
Abstract: Objective Differences between uni- and bi-nucleate mycelia of Pleurotus ostreatus were studied to aid the mushroom breeding and cultivation. Method Monokaryons MK13 and MK3 were isolated from basidiospores of P. ostreatus TD300 and mated to obtain the dikaryotic DK300. Conventional plate and fruiting tests were applied to compare the mycelial growth and fruiting body yield of the 4 strains. Genetic differences of the strains were determined by random amplified polymorphic DNA (RAPD) markers. Result There were no significant differences in mycelial growth and fruiting body yield between DK300 and TD300. However, a significant genetic difference existed between the offspring (i.e., MK13, MK3, and DK300) and parent TD300. The differences among MK13, MK3, and DK300 were minute, especially between MK3 and DK300. Conclusion Gene recombination might occur during the basidiospore formation by meiosis, but less likely in the mating of two monokaryotic strains to generate a dikaryotic offspring.
- Pleurotus ostreatus /
- monokaryon /
- dikaryon /
- yield /
- genetic diversity

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图 1 菌株在不同培养基中生长状况

Figure 1. Mycelial growth of strains in different culture media

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图 2 DNA电泳检测结果

M为λ-Hind Ⅲ。

Figure 2. DNA electrophoresis of 4 mushroom strains

M: λ-Hind III.

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图 3 随机引物扩增电泳图谱

Figure 3. PCR electrophoresis using random primers

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图 4 RAPD聚类分析

Figure 4. RAPD clustering on mushroom strains

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表 1 供试随机引物

Table 1. Random primers applied

引物编号 Primer number	引物序列（5′-3′） Primer sequence(5′−3′)
S10	CTGCTGGGAC
S22	TGCCGAGCTG
S23	AGTCAGCCAC
S28	GTGACGTAGG
S36	AGCCAGCGAA
S39	CAAACGTCGG
S43	GTCGCCGTCA
S62	GTGAGGCGTC
S75	GACGGATCAG
S78	TGAGTGGGTG
S79	GTTGCCAGCC
S124	GGTGATCAGG
S126	GGGAATTCGG

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表 2 菌株的菌丝形态

Table 2. Mycelium morphology of tested strains

菌株 Bacterial strain	菌落形态 Colonial morphology	菌丝长势 Mycelium growth vigor	菌丝颜色 Color of mycelium
TD300	半气生型 Semi-air type	+	浅白 Light white
DK300	半气生型 Semi-air type	++	白 White
MK13	匍匐型 Prostrate type	+++	白 White
MK3	匍匐型 Prostrate type	+++	白 White
+++：粗壮浓密；++：较粗壮浓密；+：纤细稀疏。 +++: thick; ++: thick and dense; +: slender sparse.

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表 3 菌株出菇情况及子实体性状

Table 3. Germination and fruiting body properties of mushrooms

菌株 Bacterial strain	出菇情况 Mushroom-budding	子实体形状 Fruit-body shape	子实体颜色 Fruit-body color
TD300	出菇 Producing mushroom	贝壳状 Conchoid type	深褐色 Dark brown
DK300	出菇 Producing mushroom	扇叶状 Fan leaf type	浅褐色 Light brown
MK13	不出菇 No mushrooms	–	–
MK3	不出菇 No mushrooms	–	–
–：不出菇。 –: No mushrooms.

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表 4 菌株出菇产量

Table 4. Yields of mushrooms

菌株 Strain	第一茬均产量 Average yield of the first crop/ （kg·袋⁻¹）	第二茬均产量 Average yield of the second crop/ （kg·袋⁻¹）	平均总产量 Average total output/ （kg·袋⁻¹）
TD300	0.273 a	0.139 a	0.412 a
DK300	0.285 a	0.114 b	0.399 a
MK13	–	–	–
MK3	–	–	–
表中不同字母表示差异显著（P＜0.05）；–：不出菇。 Different letters indicate significant difference at P＜0.05; –: no mushrooms.

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