Agronomic Characteristics and ISSR Molecular Markers of Auricularia cornea Strains
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摘要:
目的 明确主栽毛木耳菌株的亲缘关系,为毛木耳选育新品种、种质资源构建保存研究等提供参考依据。 方法 收集省内外主栽以及前期杂交的毛木耳菌株共8个,进行农艺性状以及ISSR遗传分析,农艺性状包括菌种菌丝生长速度、子实体形态特征、产量、干湿比等进行分析。 结果 供试菌株的农艺性状有一定的差异,其中菌株43012、漳耳43-28菌丝生长速度较快,耳片平均直径大,分别为11.8 、11.3 cm,耳片厚,分别为0.128 、0.126 cm,产量高,干耳总产量分别达到72.5 、71.5 g·袋−1。菌株杂10表现耳片多,菊花状。菌株CR5表现耳片腹面浅红褐色。根据晒干后毛木耳背面的色泽,将供试菌株分为白背毛木耳和黄背毛木耳两大类,其中永耳13、781、漳平42等3个菌株为黄背毛木耳,其他菌株为白背毛木耳。ISSR分子标记分析结果表明,供试菌株在相似性75%水平上,聚为5大类群,菌株781和永耳13为第一类群,2个菌株的相似性为98%,菌株CR5为第二类群,漳耳43-28、杂10为第三类群,2个菌株的相似性为76%,苏毛3号、43012为第四类群,2个菌株的相似性为83%。菌株漳平42为第五类群,漳平42与其他菌株的相似性最小为53%。 结论 结合农艺性状及ISSR聚类分析,初步判断菌株781与永耳13可能为同一品种,为同种异名现象。菌株杂10、CR5、43012、漳耳4328各有优势,可以作为杂交育种的亲本。 Abstract:Objective Morphological and genetic characteristics of strains of Auricularia cornea were studied to facilitate new variety breeding and germplasm conservation. Method The agronomic properties and ISSR molecular markers of 8 major strains of cultivated or hybrid A. cornea collected from Fujian and other producing provinces in China were determined for comparison. Result The strains had significant morphological and genetic differences. Zhanger 43-28 and Strain 43012 showed the highest rate of mycelial growth with an average diameter of 11.8 cm and 11.3 cm, respectively, a thickness of 0.128 cm and 0.126 cm, respectively, and an output of 72.5 g·bag−1 and 71.5 g·bag−1, respectively, on earpieces. Za 10 had numerous earpieces with the fruiting body a chrysanthemum-like appearance. And the ventral surface of Strain CR5 was light reddish brown in color. The dried fruiting bodies of the 8 strains were either white or yellow on the back. Thus, Yonger 13, Strain 781, and Zhangping 42 were commercially categorized as yellow-backed and the other strains white-backed A. cornea. At 75% similarity on ISSR molecular markers, the 8 strains were clustered into Group 1 that consisted of Strain 781 and Yonger 13, the similarity of the two strains was 98%. Group 2 of Strain CR5, Group 3 of Zhanger 43-28 and Za 10, the similarity of the two strains was 76%. Group 4 of Sumao 3 and Strain 43012, the similarity of the two strains was 83% and Group 5 of Zhangping 42, Zhangping 42 and the remainders at 53%. Conclusion Combined the agronomic characteristics and ISSR clustering result, Yonger 13 and Strain 781 appeared phylogenetically closely related. They were same species but had been given different names. Strains Za 10, CR5, 43012 and Zhanger 4328 have their own advantages and can be used as parents of cross breeding. -
Key words:
- Auricularia cornea /
- agronomic characters /
- ISSR /
- genetic analysis
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表 1 供试菌株
Table 1. Eight A. cornea strains studied
编号
NO.菌株名称
Strain来源
Source1 781 三明市真菌研究所
Fungi Institute of Sanming2 永耳13
Yonger13永安市耳农赠送
Yongan er nong giving3 漳耳43-28
Zhanger43-28漳州市农业科学研究所
Zhangzhou Institute of Agricultural Sciences4 苏毛3号
Sumao3江苏省农业科学院蔬菜研究所
Vegetable Research Institute, Jiangsu Academy of Agricultural Sciences5 漳平42
Zhangping42漳平市耳农赠送
Zhangping er nong giving6 43012 漳州市农业科学研究所
Zhangzhou Institute of Agricultural Sciences7 杂10
Za10漳州市农业科学研究所
Zhangzhou Institute of Agricultural Sciences8 CR5 四川省农业科学院土壤肥料研究所
Institute of Soil and Fertilizer, Sichuan Academy of Agricultural Sciences表 2 ISSR引物列表
Table 2. Sequence of ISSR primers
引物名称
Name引物序列
SequenceH3 AGAGAGAGAGAGAGAGG H4 AGAGAGAGAGAGAGAGC H5 GSGTGTGTGTGTGTGTGT H7 TCTCTCTCTCTCTCTCC H8 AGAGAGAGAGAGAGAGYA H9 AGAGAGAGAGAGAGAGYC H10 GAGAGAGAGAGAGAGAYC H11 GAGAGAGAGAGAGAGAYT H12 GAGAGAGAGAGAGAGAYG H14 TGCACACACACACAC H15 GTGACACACACACAC H16 AGTGTGTGTGTGTGT H18 GGAGTGGTGGTGGTG H19 AGAGAGAGAGAGAGAGGC H20 ACACACACACACACACCG H22 ACACACACACACACACC H23 GTGACGACTCTCTCTCT H25 ACACACACACACACACG H26 AGAGAGAGAGAGAGAGYT H27 CACACACACACACACARC H28 ACACACACACACACACYA P4 GGATGCAACACACACACAC P5 CGTGTGTGTGTGTGT P11 GAGAGAGAGAGAGAGAAC P21 AGCAGCAGCAGCAGCAGCG P26 CACCACACACACACACA 表 3 菌种菌丝生长情况
Table 3. Mycelial growth
供试菌株
Strain母种菌丝生长速度
Mycelial growth rate of
primary spawn/(mm·d−1)原种菌丝长速度
Mycelial growth rate of
secondary spawn/(mm·d−1)栽培种菌丝生长速度
Mycelial growth rate of
third spawn/(mm·d−1)781 4.10±0.015 cC 4.20±0.006 fE 4.35±0.0006 gF 永耳13
Yonger134.26±0.011 bB 4.04±0.025 gF 4.68±0.005 fE 漳耳43-28
Zhanger43-283.31±0.029 eE 4.48±0.003 bB 4.92±0.009 dC 苏毛3号
Sumao34.12±0.009 cC 4.43±0.006 dC 5.10±0.04 cB 漳平42
Zhangping423.77±0.003 dD 4.98±0.006 aA 5.25±0.011 aA 43012 4.33±0.006 aA 4.46±0.006 cBC 5.22±0.006 aA 杂10
Za102.15±0.005 gG 3.60±0.01 hG 4.80±0.044 eD CR5 3.14±0.002 fF 4.24±0.004 eD 5.15±0.04 bB 注:同列不同大写字母表示差异极显著 (P<0.01),小写字母不同表示差异显著(P<0.05),表4同.
Note: Data with different capital letters on same column indicate significant difference at P<0.01; those with different lowercase letters significant difference at P<0.05. The same as Table 4.表 4 各菌株子实体的形态特征
Table 4. Morphological characteristics of 8 A. cornea strains
菌株
Strain耳片平均直径
Average diameter/cm耳片厚度
Average diameter/cm绒毛数量
Number of villi腹面 干耳颜色
Dry color横径
Transverse diameter纵径
Longitudinal diameter棱脊
Ventral ridge背面
Back腹面
Ventral surface781 10.6±0.017 dCD 8.1±0.038 eE 0.095±0.007 cdC 少量 less 少量 less 黄褐色
yellowish brown暗紫红色
dark purple永耳13
Yonger1310.51±0.095 eD 8.2±0.01 dD 0.101±0.003 cC 少量less 多,棱形
more, prismatic浅黄褐色
light yellowish brown暗紫红色
dark purple漳耳43-28
Zhanger43-2811.3±0.05 bB 8.89±0.04 aA 0.126±0.001 aA 多more 无 none 浅灰白色
light gray white紫褐色
purplish brown苏毛3号
Sumao 311.22±0.026 bB 8.19±0.014 dD 0.125±0.003 aA 多 more 无 none 浅灰白色
light gray white紫褐色
purplish brown漳平42
Zhangping4210.5±0.015 eD 8.04±0.031 fF 0.092±0.002 dC 多 more 无 none 浅黄褐色
light yellowish brown紫褐色
purplish brown43012 11.8±0.015 aA 8.5±0.013 bB 0.128±0.006 dC 多 more 无 none 浅灰白色
light gray white紫褐色
purplish brown杂10 Za10 10.3±0.021 fE 7.8±0.01 gG 0.113±0.001 bB 多 more 多,网状
more, reticular浅灰色
light grey紫褐色
purplish brownCR5 10.7±0.086 cC 8.3±0.007 cC 0.122±0.003 aA 少量 less 少量 less 白色 white 浅红褐色
light reddish brown表 5 各菌株产量情况表
Table 5. Average yields of 8 A. cornea strains
菌株
Strain第一潮平均产量
First average production/
(g·袋−1)第二潮平均产量
Second average production/
(g·袋−1)总产量
Total production/
(g·袋−1)5%显著水平
Significance level第一潮产量/总产量
First average production/
Total production781 61.0 6.1 67.1 cd 90.9% 永耳13
Yonger1361.5 7.5 69.0 c 89.13% 漳耳43-28
Zhanger43-2866.5 5.0 71.5 ab 93.01% 苏毛3号
Sumao 365.0 5.2 70.2 b 92.59% 漳平42
Zhangping4255.0 7.5 62.5 e 88.00% 43012 63.5 9.0 72.5 a 87.59% 杂10
Za1062.5 5.0 67.5 cd 92.59% CR5 57.5 8.5 66.0 d 87.12% 表 6 不同菌株子实体干湿比分析
Table 6. Dry to wet fruiting body weight ratio of 8 A. cornea strains
菌株
Strain毛木耳干湿比
Dry and wet ratio1.5 h 3 h 18 h 20 h 781 4.70 5.41 6.00 6.02 永耳13 Yonger13 4.73 5.47 6.06 6.07 漳耳43-28 Zhanger43-28 4.89 5.2 5.51 5.54 苏毛3号 Sumao No.3 4.81 5.13 5.5 5.52 漳平42 Zhangping42 4.63 5.03 5.39 5.39 43012 4.62 5.06 5.52 5.53 杂10 Za10 5.32 5.68 6.34 6.33 CR5 3.65 3.76 3.96 3.95 -
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