Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (10): 26-33.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0588
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LI E1(
), HUANG Yong1, MENG Yuan-yuan1, LI Xuan2, DU Guang-hui2(), LIU Fei-hu1,2()
Received:2020-05-14
Online:2021-10-26
Published:2021-11-12
Contact:
DU Guang-hui,LIU Fei-hu
E-mail:2049829995@qq.com;dgh2012@ynu.edu.cn;dmzpynu@126.com
LI E, HUANG Yong, MENG Yuan-yuan, LI Xuan, DU Guang-hui, LIU Fei-hu. Isolation and Identification of the Endophytic Fungi of‘Bama hemp’ Under Salt Stress and Its Diversity Analysis[J]. Biotechnology Bulletin, 2021, 37(10): 26-33.
Fig. 1 Sterilization effect of endophytic fungi A, B and C respectively refers to tissue compression test,coating test and ultra clean bench test
| 盐浓度Salt concentration /（mmol·L-1） | 科Families | 属Genera | 种Species |
|---|---|---|---|
| 0 | Xylariaceae炭角科,Chaetomiaceae 毛壳科,Pleosporaceae多孢菌科,Cladosporiaceae枝孢菌科,Trichosphaeriaceae毛球藻科,Hypocreales incertae sedis,Diaporthaceae,Nectriaceae从赤壳科,Glomerellaceae小从赤壳科,Hypocreaceae竹盖菌科（9） | Annulohypoxylon炭团菌属,Nigrospora 黑孢霉属,Chaetomium毛壳菌属,Cladosporium 枝孢霉属,Nemania炭垫菌属,Alternaria链格孢属,Aphanocladium丝枝霉属,Colletotrichum炭疽病属,Diaporth茎点霉属,Fusarium镰孢菌属,Neocosmospora炭疽病属,Pleosporales格孢腔菌,Hypoxylon炭团菌属,Trichoderma木霉菌属（14） | Diaporthe novem,Nemania diffusa②,Alternaria sp. R60.1,Annulohypoxylon bovei,Cladosporium sp②.,Annulohypoxylon stygium,Aphanocladium album,Pleosporales sp. 9 MSP-2018,Colletotrichum kahawae,Neocosmospora rubicola,Hypoxylon fragiforme,Nigrospora oryzae,Trichoderma harzianum,Nigrospora sphaerica,Annulohypoxylon atroroseum,Fusarium oxysporum②,Chaetomium globosum③（22） |
| 100 | Xylariaceae炭角科,Chaetomiaceae 毛壳科,Pleosporaceae多孢菌科,Cladosporiaceae枝孢菌科,Trichosphaeriaceae毛球藻科,Hypocreales incertae sedis,Nectriaceae从赤壳科,Hypocreaceae竹盖菌科,Xylariales incertae sedis（9） | Annulohypoxylon炭团菌属,Nigrospora 黑孢霉属,Chaetomium毛壳菌属,Cladosporium 枝孢霉属,Nemania炭垫菌属,Alternaria链格孢属,Aphanocladium丝枝霉属,Fusarium镰孢菌属,Neocosmospora炭疽病属,Hypoxylon炭团菌属,Trichoderma木霉菌属,Hansfordia汉斯霉属,Kretzschmaria炭墩属（13） | Nigrospora sphaerica,Nemania diffusa,Nigrospora oryzae,Annulohypoxylon stygium,Aphanocladium album,Hansfordia sp.,Kretzschmaria pavimentosa,Alternaria sp.,Fusarium equiseti②,Neocosmospora rubicola,Trichoderma harzianum②,Fusarium solani,Chaetomium globosum②,Hypoxylon pilgerianum,Alternaria macrospora,Cladosporium sp.,Hypoxylon trugodes（20） |
| 200 | Xylariaceae炭角科,Chaetomiaceae 毛壳科,Cladosporiaceae枝孢菌科,Nectriaceae从赤壳科,Hypocreaceae竹盖菌科,Xylariales incertae sedis（6） | Annulohypoxylon炭团菌属,Chaetomium毛壳菌属,Cladosporium 枝孢霉属,Nemania炭垫菌属,Fusarium镰孢菌属,Trichoderma木霉菌属,Hansfordia汉斯霉属,Diaporth茎点霉属（8） | Nemania sp. AX40,Diaporthe novem,Nemania diffusa,Annulohypoxylon stygium,Fusarium sp.,Hansfordia sp. JP38-6,Chaetomium globosum,Trichoderma harzianum,Cladosporium sp.,Fusarium equiseti,Fusarium oxysporum（11） |
| 300 | Xylariaceae炭角科,Chaetomiaceae 毛壳科,Pleosporaceae多孢菌科,Cladosporiaceae枝孢菌科,Trichosphaeriaceae毛球藻科,Diaporthaceae,Glomerellaceae小从赤壳科,Hypocreaceae竹盖菌科,Leptosphaeriaceae小球腔菌科,Ceratobasidiaceae（9） | Annulohypoxylon炭团菌属,Nigrospora 黑孢霉属,Chaetomium毛壳菌属,Cladosporium 枝孢霉属,Alternaria链格孢属,Colletotrichum炭疽病属,Diaporth茎点霉属,Fusarium镰孢菌属,Pleosporales格孢腔菌,Hypoxylon炭团菌属,Trichoderma木霉菌属,Leptosphaeria小球腔菌属（13） | Diaporthe novem②,Nemania diffusa,Leptosphaeria microscopica,Annulohypoxylon stygium,Nigrospora oryzae,Fusarium sp.,Pleosporales sp. 9 MSP-2018,Alternaria sp,Annulohypoxylon atroroseum,Hypoxylon fragiforme,Colletotrichum sp.,Trichoderma harzianum,Chaetomium globosum②,Cladosporium sp.,Fusarium equiseti,Fusarium oxysporum②,Thanatephorus cucumeris（20） |
Table 1 Comparison of species of endophytic fungi isolated from “Bama hemp” under different concentrations of salt stress
| 盐浓度Salt concentration /（mmol·L-1） | 科Families | 属Genera | 种Species |
|---|---|---|---|
| 0 | Xylariaceae炭角科,Chaetomiaceae 毛壳科,Pleosporaceae多孢菌科,Cladosporiaceae枝孢菌科,Trichosphaeriaceae毛球藻科,Hypocreales incertae sedis,Diaporthaceae,Nectriaceae从赤壳科,Glomerellaceae小从赤壳科,Hypocreaceae竹盖菌科（9） | Annulohypoxylon炭团菌属,Nigrospora 黑孢霉属,Chaetomium毛壳菌属,Cladosporium 枝孢霉属,Nemania炭垫菌属,Alternaria链格孢属,Aphanocladium丝枝霉属,Colletotrichum炭疽病属,Diaporth茎点霉属,Fusarium镰孢菌属,Neocosmospora炭疽病属,Pleosporales格孢腔菌,Hypoxylon炭团菌属,Trichoderma木霉菌属（14） | Diaporthe novem,Nemania diffusa②,Alternaria sp. R60.1,Annulohypoxylon bovei,Cladosporium sp②.,Annulohypoxylon stygium,Aphanocladium album,Pleosporales sp. 9 MSP-2018,Colletotrichum kahawae,Neocosmospora rubicola,Hypoxylon fragiforme,Nigrospora oryzae,Trichoderma harzianum,Nigrospora sphaerica,Annulohypoxylon atroroseum,Fusarium oxysporum②,Chaetomium globosum③（22） |
| 100 | Xylariaceae炭角科,Chaetomiaceae 毛壳科,Pleosporaceae多孢菌科,Cladosporiaceae枝孢菌科,Trichosphaeriaceae毛球藻科,Hypocreales incertae sedis,Nectriaceae从赤壳科,Hypocreaceae竹盖菌科,Xylariales incertae sedis（9） | Annulohypoxylon炭团菌属,Nigrospora 黑孢霉属,Chaetomium毛壳菌属,Cladosporium 枝孢霉属,Nemania炭垫菌属,Alternaria链格孢属,Aphanocladium丝枝霉属,Fusarium镰孢菌属,Neocosmospora炭疽病属,Hypoxylon炭团菌属,Trichoderma木霉菌属,Hansfordia汉斯霉属,Kretzschmaria炭墩属（13） | Nigrospora sphaerica,Nemania diffusa,Nigrospora oryzae,Annulohypoxylon stygium,Aphanocladium album,Hansfordia sp.,Kretzschmaria pavimentosa,Alternaria sp.,Fusarium equiseti②,Neocosmospora rubicola,Trichoderma harzianum②,Fusarium solani,Chaetomium globosum②,Hypoxylon pilgerianum,Alternaria macrospora,Cladosporium sp.,Hypoxylon trugodes（20） |
| 200 | Xylariaceae炭角科,Chaetomiaceae 毛壳科,Cladosporiaceae枝孢菌科,Nectriaceae从赤壳科,Hypocreaceae竹盖菌科,Xylariales incertae sedis（6） | Annulohypoxylon炭团菌属,Chaetomium毛壳菌属,Cladosporium 枝孢霉属,Nemania炭垫菌属,Fusarium镰孢菌属,Trichoderma木霉菌属,Hansfordia汉斯霉属,Diaporth茎点霉属（8） | Nemania sp. AX40,Diaporthe novem,Nemania diffusa,Annulohypoxylon stygium,Fusarium sp.,Hansfordia sp. JP38-6,Chaetomium globosum,Trichoderma harzianum,Cladosporium sp.,Fusarium equiseti,Fusarium oxysporum（11） |
| 300 | Xylariaceae炭角科,Chaetomiaceae 毛壳科,Pleosporaceae多孢菌科,Cladosporiaceae枝孢菌科,Trichosphaeriaceae毛球藻科,Diaporthaceae,Glomerellaceae小从赤壳科,Hypocreaceae竹盖菌科,Leptosphaeriaceae小球腔菌科,Ceratobasidiaceae（9） | Annulohypoxylon炭团菌属,Nigrospora 黑孢霉属,Chaetomium毛壳菌属,Cladosporium 枝孢霉属,Alternaria链格孢属,Colletotrichum炭疽病属,Diaporth茎点霉属,Fusarium镰孢菌属,Pleosporales格孢腔菌,Hypoxylon炭团菌属,Trichoderma木霉菌属,Leptosphaeria小球腔菌属（13） | Diaporthe novem②,Nemania diffusa,Leptosphaeria microscopica,Annulohypoxylon stygium,Nigrospora oryzae,Fusarium sp.,Pleosporales sp. 9 MSP-2018,Alternaria sp,Annulohypoxylon atroroseum,Hypoxylon fragiforme,Colletotrichum sp.,Trichoderma harzianum,Chaetomium globosum②,Cladosporium sp.,Fusarium equiseti,Fusarium oxysporum②,Thanatephorus cucumeris（20） |
Fig. 2 Relative abundance of endophytic fungi under diff-erent salt treatments
| 菌株编号 Strain code | 鉴定结果 Identification results | 中文名 Chinese name | 登录号 Login number | 相似度 Similarity/% |
|---|---|---|---|---|
| B17,B51,B58 | Nemania diffusa | 炭垫菌 | MK247796.1 | 100 |
| B26,B19,B33,B52,B23,B60 | Annulohypoxylon stygium | 炭团菌 | FJ848865.1 | 100 |
| B44,B37,B56,B62,B16,B18 | Chaetomium globosum | 球毛壳菌 | KF156299.1 | 100 |
| B10,B70,B67,B65 | Cladosporium sp. | 枝孢霉 | MK268136.1 | 99.80 |
| B96,B101,B91,B86,B95,B90,B76,B93 | Trichoderma harzianum | 哈茨木霉 | KT323178.2 | 100 |
Table 2 Endophytic fungi and its sequence analysis of ‘Bama hemp’ under different concentrations of salt stress
| 菌株编号 Strain code | 鉴定结果 Identification results | 中文名 Chinese name | 登录号 Login number | 相似度 Similarity/% |
|---|---|---|---|---|
| B17,B51,B58 | Nemania diffusa | 炭垫菌 | MK247796.1 | 100 |
| B26,B19,B33,B52,B23,B60 | Annulohypoxylon stygium | 炭团菌 | FJ848865.1 | 100 |
| B44,B37,B56,B62,B16,B18 | Chaetomium globosum | 球毛壳菌 | KF156299.1 | 100 |
| B10,B70,B67,B65 | Cladosporium sp. | 枝孢霉 | MK268136.1 | 99.80 |
| B96,B101,B91,B86,B95,B90,B76,B93 | Trichoderma harzianum | 哈茨木霉 | KT323178.2 | 100 |
| 共有菌名称 Name of common endophytic fungi | CK | 100 mmol/L | 200 mmol/L | 300 mmol/L | 总计 Total | ||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 上 叶 | 中 叶 | 下 叶 | 上 茎 | 中 茎 | 下 茎 | 根 | 上 叶 | 中 叶 | 下 叶 | 上 茎 | 中 茎 | 下 茎 | 根 | 上 叶 | 中 叶 | 下 叶 | 上 茎 | 中 茎 | 下 茎 | 根 | 上 叶 | 中叶 | 下 叶 | 上茎 | 中茎 | 下茎 | 根 | ||
| Nemania diffusa | — | 1 | — | — | — | — | — | — | 1 | 2 | — | — | — | — | — | 1 | — | — | — | — | — | — | 1 | — | — | — | — | 6 | |
| Annulohypoxylon stygium | — | — | 4 | — | — | — | — | — | 1 | 1 | — | — | — | — | — | 15 | 1 | — | — | — | — | — | — | 1 | — | — | — | — | 23 |
| Chaetomium globosum | — | 1 | 4 | — | — | — | — | — | 2 | 5 | — | — | — | — | — | — | 4 | — | — | — | — | — | — | 1 | — | — | — | — | 17 |
| Cladosporium sp. | — | 2 | 8 | 6 | — | 1 | — | — | 5 | 5 | 2 | 6 | 1 | — | 2 | — | 2 | 4 | 6 | 3 | 0 | 3 | 2 | 5 | — | 5 | 4 | — | 112 |
| Trichoderma harzianum | — | — | — | — | — | — | 50 | — | — | — | — | — | 8 | 18 | — | — | — | — | — | 2 | 30 | — | — | — | — | — | 7 | 46 | 161 |
Table 3 Number of endophytic fungi isolated from ‘Bama hemp’ under different concentrations of salt stress
| 共有菌名称 Name of common endophytic fungi | CK | 100 mmol/L | 200 mmol/L | 300 mmol/L | 总计 Total | ||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 上 叶 | 中 叶 | 下 叶 | 上 茎 | 中 茎 | 下 茎 | 根 | 上 叶 | 中 叶 | 下 叶 | 上 茎 | 中 茎 | 下 茎 | 根 | 上 叶 | 中 叶 | 下 叶 | 上 茎 | 中 茎 | 下 茎 | 根 | 上 叶 | 中叶 | 下 叶 | 上茎 | 中茎 | 下茎 | 根 | ||
| Nemania diffusa | — | 1 | — | — | — | — | — | — | 1 | 2 | — | — | — | — | — | 1 | — | — | — | — | — | — | 1 | — | — | — | — | 6 | |
| Annulohypoxylon stygium | — | — | 4 | — | — | — | — | — | 1 | 1 | — | — | — | — | — | 15 | 1 | — | — | — | — | — | — | 1 | — | — | — | — | 23 |
| Chaetomium globosum | — | 1 | 4 | — | — | — | — | — | 2 | 5 | — | — | — | — | — | — | 4 | — | — | — | — | — | — | 1 | — | — | — | — | 17 |
| Cladosporium sp. | — | 2 | 8 | 6 | — | 1 | — | — | 5 | 5 | 2 | 6 | 1 | — | 2 | — | 2 | 4 | 6 | 3 | 0 | 3 | 2 | 5 | — | 5 | 4 | — | 112 |
| Trichoderma harzianum | — | — | — | — | — | — | 50 | — | — | — | — | — | 8 | 18 | — | — | — | — | — | 2 | 30 | — | — | — | — | — | 7 | 46 | 161 |
Fig. 3 Diversity analysis of endophytic fungi in hemp under different concentrations of salt treatment Small letters a,b and c indicate significant difference（P < 0.05）. The same below
| 盐浓度Salt concentr ation/（mmol·L-1） | 0 | 100 | 200 | 300 |
|---|---|---|---|---|
| 0 | ||||
| 100 | 0.36±0.12a | |||
| 200 | 0.30±0.08a | 0.20±0.13a | ||
| 300 | 0.33±0.06a | 0.17±0.17a | 0.35±0.21a |
Table 4 Comparison of similarity coefficient of hemp end-ophytic fungi among different salt concentrations
| 盐浓度Salt concentr ation/（mmol·L-1） | 0 | 100 | 200 | 300 |
|---|---|---|---|---|
| 0 | ||||
| 100 | 0.36±0.12a | |||
| 200 | 0.30±0.08a | 0.20±0.13a | ||
| 300 | 0.33±0.06a | 0.17±0.17a | 0.35±0.21a |
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