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作物杂志,2024, 第6期: 194–204 doi: 10.16035/j.issn.1001-7283.2024.06.026

• 生理生化·植物营养·栽培耕作 • 上一篇    下一篇

燕麦地上部可培养内生真菌多样性分析及其功能研究

李晓婷1(), 张婷婷2, 张艳丽1, 李志伟1, 韩丽1, 赵鑫瑶1, 张永平1, 李立军1()   

  1. 1内蒙古农业大学农学院，010019，内蒙古呼和浩特
    2乌兰察布市农林科学研究所，012000，内蒙古乌兰察布
  • 收稿日期:2023-07-28 修回日期:2023-10-31 出版日期:2024-12-15 发布日期:2024-12-05
  • 通讯作者: 李立军，研究方向为寒旱区农作制与农业生态，E-mail：imaullj@163.com
  • 作者简介:李晓婷，研究方向为农牧交错区农作制与农业生态，E-mail：lixt1229@163.com
  • 基金资助:
    内蒙古自治区科技计划项目(2022YFXZ0031);呼和浩特市科技计划项目(2020-农-1)

Diversity Analysis and Function Study of Culturable Endophytic Fungi in Oat Shoot

Li Xiaoting1(), Zhang Tingting2, Zhang Yanli1, Li Zhiwei1, Han Li1, Zhao Xinyao1, Zhang Yongping1, Li Lijun1()   

  1. 1Agricultural College of Inner Mongolia Agricultural University, Hohhot 010019, Inner Mongolia, China
    2Ulanqab Institute of Agriculture and Forestry Sciences, Ulanqab 012000, Inner Mongolia, China
  • Received:2023-07-28 Revised:2023-10-31 Online:2024-12-15 Published:2024-12-05

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摘要：

为探究燕麦叶和茎中可培养内生真菌分布情况、群落组成及其功能特性，通过组织培养分离法对阴山北麓区燕麦拔节期叶片和茎的内生真菌分离纯化，利用ITS鉴定并分析其多样性，同时测定其溶磷、解钾和分泌生长素（IAA）能力。结果发现，燕麦的叶和茎中分离得到74株内生真菌，总定殖率为61.94%，总分离率为20.56%，经过分子鉴定属于2门12属，其中优势菌门为子囊菌门（Ascomycota），优势菌属为链格孢菌属（Alternaria，分离率=5.56%，分离频率=27.03%）和镰刀菌属（Fusarium，分离率=8.06%，分离频率=39.19%）。多样性分析发现，茎中分离的内生真菌多样性高于叶，其多样性指数和辛普森指数分别为3.51和0.78。相似性分析发现，叶和茎中内生真菌的相似系数为0.15。对分离得到的内生真菌的功能分析发现，79.73%的菌株具有溶磷能力，72.97%的菌株具有解钾能力，40.54%的菌株能够分泌IAA。根据内生真菌的功能将74株内生真菌分为4类，分别有41、5、21和7株菌株，最优第4类菌株大部分均具有溶磷、解钾能力，且分泌IAA的能力最强。燕麦地上部组织中的内生真菌物种较为丰富，可为内生真菌的应用提供菌种资源，不同内生真菌的功能差异较大，功能性较强菌株在农业生产中可能会发挥其潜在促生长特性。

关键词: 燕麦, 内生真菌, 多样性, 溶磷, 解钾, 生长素

Abstract:

In order to investigate the distribution, community composition, and functional characteristics of culturable endophytic fungi leaves and stems of oats (Avena sativa L.), the endophytic fungi in leaves and stems of oats at the jointing stage in the northern foothills of Yinshan were isolated and purified by the tissue culture isolation method. Their diversity was identified and analyzed by using ITS, and the abilities of phosphate- solubilizing and potassium-solubilizing, secreting auxin (IAA) were measured. The results showed that a total of 74 endophytic fungi were isolated from the leaves and stems of oats, with the total colonization rate and the total isolation rate of 61.94% and 20.56%, respectively, they were identified as belonging to two phyla and 12 genera by molecular identification, among which Ascomycota was the dominant phylum, and Alternaria (isolation rate=5.56%, isolation frequency=27.03%) and Fusarium (isolation rate=8.06%, isolation frequency=39.19%) were the dominant species. Diversity analysis showed that the diversity of endophytic fungi isolated from stems was higher than those from leaves, with the diversity index and Simpson index being 3.51 and 0.78, respectively. Similarity analysis showed that the similarity coefficient of endophytic fungi in leaves and stems was 0.15. The functional analysis of the isolated endophytic fungi showed that 79.73% of the strains had phosphate- solubilizing, 72.97% had potassium-solubilizing, and 40.54% could secrete IAA. According to the function of endophytic fungi, 74 strains of endophytic fungi were divided into four categories, including 41, 5, 21 and 7 strains, respectively. Most of the optimal strains of the 4th category had the ability to dissolve phosphorus and potassium, and the ability to secrete IAA was the highest. There are abundant endophytic fungi species in the leaves and stems of oat, which would provide strain resources for the application of endophytic fungi. The functions of different endophytic fungi are quite distinct, and the strains with stronger functions may exert their potential growth-promoting characteristics in agricultural production, which is beneficial to the sustainable development of agriculture.

Key words: Oat, Endophytic fungi, Diversity, Phosphate-solubilizing, Potassium-solubilizing, IAA

表1

燕麦内生真菌序列鉴定

菌株
Strain		 登录号
GenBank ID		 来源
Source		 DNA鉴定结果
DNA identification result		 相似性
Identity (%)		 菌株
Strain		 登录号
GenBank ID		 来源
Source		 DNA鉴定结果
DNA identification result		 相似性
Identity (%)
OL-1 ON495971 叶 链格孢菌 99.81 OL-38 ON500580 叶 木贼镰孢菌 100.00
OL-2 ON495943 叶 木贼镰孢菌 99.61 OL-39 ON500581 叶 链格孢菌 100.00
OL-3 ON495944 叶 链格孢菌 100.00 OL-40 ON500582 叶 真菌 99.40
OL-4 ON495945 叶 互隔链格孢 99.44 OL-41 ON500583 叶 子囊菌 99.60
OL-5 ON495946 叶 木贼镰孢菌 99.41 OL-42 ON500584 叶 木贼镰孢菌 99.80
OL-6 ON495947 叶 木贼镰孢菌 100.00 OL-43 ON500585 叶 木贼镰孢菌 99.61
OL-7 ON495948 叶 内生真菌 99.63 OL-44 ON500586 叶 互隔链格孢 99.81
OL-8 ON495949 叶 木贼镰孢菌 99.61 OL-45 ON500587 叶 木贼镰孢菌 99.61
OS-9 ON495950 茎 木贼镰孢菌 99.81 OS-46 ON500588 茎 紧密帚枝霉 99.45
OL-10 ON495951 叶 木贼镰孢菌 99.22 OS-47 ON500589 茎 裂褶菌 99.50
OL-11 ON500553 叶 木贼镰孢菌 99.22 OL-48 ON500590 叶 细极链格孢 99.44
OS-12 ON500554 茎 互隔链格孢 99.26 OL-49 ON500591 叶 互隔链格孢 99.63
OL-13 ON500555 叶 木贼镰孢菌 100.00 OS-50 ON500592 茎 芸苔链格孢 99.26
OL-14 ON500556 叶 木贼镰孢菌 99.41 OL-51 ON500593 叶 木贼镰孢菌 99.80
OS-15 ON500557 茎 木贼镰孢菌 100.00 OL-52 ON500594 叶 子囊菌 99.40
OS-16 ON500558 茎 互隔链格孢 99.63 OL-53 ON500595 叶 互隔链格孢 100.00
OS-17 ON500559 茎 芸苔链格孢 99.81 OL-54 ON500596 叶 木贼镰孢菌 99.41
OS-18 ON500560 茎 派伦霉 99.21 OL-55 ON500597 叶 木贼镰孢菌 99.61
OS-19 ON500561 茎 茎点霉 99.60 OL-56 ON500598 叶 木贼镰孢菌 99.41
OL-20 ON500562 叶 派伦霉 99.21 OL-57 ON500599 叶 木贼镰孢菌 99.41
OS-21 ON500563 茎 高粱附球菌 99.40 OL-58 ON500600 叶 木贼镰孢菌 99.80
OS-22 ON500564 茎 互隔链格孢 99.44 OL-59 ON500601 叶 木贼镰孢菌 99.03
OS-23 ON500565 茎 高粱附球菌 99.60 OL-60 ON500602 叶 互隔链格孢 99.63
OL-24 ON500566 叶 子囊菌 99.40 OS-61 ON500603 茎 廯囊腔菌 99.61
OS-25 ON500567 茎 雪球微座孢 99.42 OL-62 ON500604 叶 木贼镰孢菌 99.61
OS-26 ON500568 茎 派伦霉 99.40 OL-63 ON500605 叶 木贼镰孢菌 99.41
OS-27 ON500569 茎 小双孢腔菌 99.40 OL-64 ON500606 叶 木贼镰孢菌 99.61
OS-28 ON500570 茎 镰刀菌属 99.61 OS-65 ON500607 茎 互隔链格孢 99.45
OS-29 ON500571 茎 派伦霉 99.21 OS-66 ON500608 茎 链格孢菌 100.00
OS-30 ON500572 茎 春孔菌 99.62 OL-67 ON500609 叶 木贼镰孢菌 99.61
OS-31 ON500573 茎 链格孢菌 99.44 OL-68 ON500610 叶 子囊菌 99.40
OS-32 ON500574 茎 木贼镰孢菌 99.61 OS-69 ON500611 茎 廯囊腔菌 99.61
OL-33 ON500575 叶 小双孢腔菌 99.60 OS-70 ON500612 茎 雪球微座孢 99.23
OL-34 ON500576 叶 木贼镰孢菌 99.61 OS-71 ON500613 茎 紧密帚枝霉 99.45
OL-35 ON500577 叶 木贼镰孢菌 99.42 OL-72 ON500614 叶 链格孢菌 100.00
OL-36 ON500578 叶 子囊菌 99.40 OL-73 ON500615 叶 芸苔链格孢 99.81
OL-37 ON500579 叶 澳大利亚小光壳 99.20 OS-74 ON500616 茎 互隔链格孢 99.44

表2

燕麦内生真菌群落组成

门Phylum 纲Class 目Order 科Family 属Genus 种Specie N IR (%) IF (%)
Ascomycota子囊菌门 座囊菌纲 格孢腔菌目 格孢菌科 链格孢属 互隔链格孢 10 2.78 13.51
芸苔链格孢 3 0.83 4.05
链格孢菌 5 1.39 6.76
链格孢菌 1 0.28 1.35
细极链格孢 1 0.28 1.35
亚隔孢壳科 亚隔孢壳属 小双胞腔菌 2 0.56 2.70
附球菌属 高粱附球菌 2 0.56 2.70
茎点霉属 茎点霉 1 0.28 1.35
小光壳属 澳大利亚小光壳 1 0.28 1.35
派伦霉属 派伦霉 2 0.56 2.70
Peyronellaea sp.派伦霉 2 0.56 2.70
子囊菌纲 肉座菌目 丛赤壳科 镰刀菌属 木贼镰孢菌 28 7.78 37.84
镰刀菌 1 0.28 1.35
Sarocladiaceae 帚枝霉属 紧密帚枝霉 2 0.56 2.70
炭角菌目 微座囊菌科 微座孢属 Microdochium bolleyi 2 0.56 2.70
小丛壳目 小不整球囊菌科 小不整球壳属 癣囊腔菌 2 0.56 2.70
未识别的子囊菌 子囊菌 5 1.39 6.76
Basidiomycota担子菌门 伞菌纲 多孔菌目 多孔菌科 春孔菌属 春孔菌 1 0.28 1.35
伞菌目 裂褶菌科 裂褶菌属 裂褶菌 1 0.28 1.35
Unclassified_fungi未识别的真菌 2 0.56 2.70
总计Total 74 20.56 100.00

表3

燕麦内生真菌定殖率和分离率

项目Index 叶Leaf 茎Stem
培养组织块数（Nt）Number of tissue blocks 180 180
有菌组织块数（N0）Number of bacterial tissue blocks 163 160
分离菌株数（Ni）Number of isolated strains 46 28
定殖率（CR）Colonization rate (%) 90.56 88.89
分离率（IR）Isolation rate (%) 28.22 17.50

图1

燕麦不同部位内生真菌分离频率

表4

燕麦不同部位内生真菌多样性指数和相似性指数

项目Index 叶
Leaf		 茎
Stem		 内生真菌
Endophytic
fungi
物种数（S）Number of species 5 9 12
卡玛戈指数Camargo’s index 0.20 0.11 0.083
多样性指数H’
Shannon-Weiner diversity index		 2.00 3.51 1.88
均匀度指数E（ACE） 1.03 1.46 0.71
丰富度指数R（Chao1） 1.57 1.80 3.02
辛普森指数D（Simpson） 0.62 0.78 0.76
相似性指数Cs
Sorenson’s similarity coefficient		 叶 － －
茎 0.15 －

表5

燕麦内生真菌功能的分类

类群
Group		 菌株
Strain		 解有机磷能力
Ability of organic phosphate-solubilizing		 解无机磷能力
Ability of inorganic phosphate-solubilizing		 解钾能力
Ability of potassium-solubilizing		 IAA含量
IAA content
Ⅰ OL-1 21.61±2.21 23.49±1.65 17.50±1.32 0.00±0.00
OL-6 4.34±0.49 33.05±1.35 10.73±1.31 0.00±0.00
OS-9 2.01±0.27 48.97±1.21 0.00±0.00 0.00±0.00
OL-11 0.00±0.00 41.29±0.66 6.00±0.26 0.00±0.00
OL-13 1.38±0.24 26.18±1.55 0.00±0.00 0.00±0.00
OL-14 1.67±0.93 18.30±1.86 1.60±0.60 0.00±0.00
OS-15 1.51±0.84 45.07±4.86 11.05±0.93 0.00±0.00
OS-16 2.66±0.12 27.28±1.37 28.67±1.29 0.00±0.00
OS-21 7.37±0.90 19.65±0.60 0.00±0.00 0.00±0.00
OS-22 14.75±3.93 28.13±1.58 0.00±0.00 0.00±0.00
OS-23 13.57±1.05 20.39±1.68 0.00±0.00 0.00±0.00
OL-24 7.18±0.52 0.00±0.00 0.00±0.00 0.00±0.00
OS-26 5.56±0.27 23.54±2.78 19.13±0.35 0.00±0.00
OS-27 6.23±0.54 16.41±0.58 16.55±1.27 0.00±0.00
OS-29 9.37±0.34 20.89±1.59 33.77±0.90 0.00±0.00
OS-31 12.21±2.75 34.29±2.72 11.20±0.42 0.00±0.00
OS-32 1.93±0.16 41.59±3.93 4.10±0.66 0.00±0.00
OL-33 5.23±0.35 12.38±0.56 0.00±0.00 0.00±0.00
OL-34 4.20±0.28 21.46±0.94 20.10±0.57 0.00±0.00
OL-35 7.17±0.34 37.22±2.72 3.47±0.57 0.00±0.00
OL-36 10.70±0.46 0.00±0.00 0.00±0.00 0.00±0.00
OL-37 12.95±2.11 18.12±0.99 10.27±0.36 0.00±0.00
OL-38 4.97±0.23 28.50±2.47 0.00±0.00 0.00±0.00
OL-40 1.66±0.22 18.76±1.62 0.00±0.00 0.00±0.00
OL-41 5.31±0.12 0.00±0.00 6.23±0.21 0.00±0.00
OL-43 0.00±0.00 19.38±1.51 1.63±0.29 0.00±0.00
OL-44 9.67±0.79 9.46±0.37 0.00±0.00 0.00±0.00
OL-45 1.13±0.26 20.08±2.95 22.40±2.93 0.00±0.00
OS-47 1.47±0.13 22.93±1.94 6.15±0.21 0.00±0.00
OL-48 5.14±0.93 18.85±1.02 24.37±5.40 0.00±0.00
OL-49 6.69±0.81 24.13±1.84 33.40±2.55 0.00±0.00
OL-52 6.87±0.15 32.84±3.12 0.00±0.00 0.00±0.00
OL-58 4.79±0.24 0.00±0.00 0.00±0.00 0.00±0.00
OL-59 1.57±0.53 28.86±1.83 0.00±0.00 0.00±0.00
OS-61 0.00±0.00 47.13±3.58 0.00±0.00 13.51±1.26
OL-64 0.00±0.00 24.07±1.22 8.85±0.46 0.00±0.00
OL-68 9.71±0.25 17.39±0.73 7.77±0.11 0.00±0.00
OS-70 15.67±0.41 24.62±1.31 0.00±0.00 0.00±0.00
OS-71 9.85±0.40 30.84±3.74 4.57±0.38 0.00±0.00
OL-73 9.61±0.94 24.57±1.09 12.90±1.84 0.00±0.00
OS-74 0.00±0.00 28.50±2.12 14.90±1.98 0.00±0.00
Ⅱ OL-4 10.95±1.33 27.37±2.06 53.33±1.25 21.12±0.26
OL-5 3.39±0.43 29.58±1.09 43.00±2.13 0.00±0.00
OS-25 4.57±0.26 13.75±0.42 51.77±1.89 0.00±0.00
OS-28 2.42±0.61 47.13±1.93 25.90±0.71 0.00±0.00
OL-62 1.32±0.18 40.01±2.87 40.67±1.76 0.00±0.00
Ⅲ OL-2 3.41±0.69 27.97±0.97 31.27±2.62 53.33±1.22
OL-3 13.17±2.81 20.38±1.12 13.33±2.32 37.68±1.88
OS-12 6.24±0.54 24.93±1.18 26.37±1.78 49.36±0.46
OS-17 8.06±0.21 16.09±0.11 12.80±1.47 53.98±1.85
OS-18 5.05±0.11 18.10±0.74 46.27±2.87 57.22±0.62
OS-19 14.81±0.28 21.47±1.99 27.37±2.95 61.18±1.89
OL-20 13.82±4.57 0.00±0.00 0.00±0.00 58.75±0.88
OS-30 0.00±0.00 41.28±2.84 0.00±0.00 47.97±1.29
OL-42 4.84±0.28 32.87±0.64 34.25±2.33 60.75±2.19
OL-51 8.97±0.25 22.42±1.69 22.14±3.37 38.32±0.10
OL-53 10.40±0.61 23.62±2.43 18.27±2.83 60.05±2.06
OL-54 5.88±0.17 28.72±1.22 12.62±0.21 65.99±3.35
OL-55 0.00±0.00 24.34±1.59 10.45±1.06 47.46±1.38
OL-56 2.05±0.25 19.13±1.13 8.70±0.14 50.61±0.44
OL-57 1.09±0.09 0.00±0.00 31.73±2.75 75.26±2.24
OL-60 5.73±0.27 22.60±1.46 59.37±3.16 53.99±0.24
OL-63 7.25±0.13 26.34±0.89 20.30±3.56 49.52±0.89
OS-65 9.69±0.35 22.23±0.46 0.00±0.00 57.01±0.53
OL-67 5.87±0.18 33.27±2.07 11.90±1.62 54.09±1.58
OS-69 1.56±0.17 28.59±1.95 33.63±2.61 41.19±0.53
OL-72 6.63±0.78 24.77±1.73 17.00±1.75 40.48±1.18
Ⅳ OL-7 8.49±0.43 20.28±0.19 18.05±0.43 89.45±5.13
OL-8 3.61±0.15 28.53±0.97 11.65±0.35 81.75±1.88
OL-10 0.00±0.00 42.07±0.62 33.30±1.61 96.61±2.15
OL-39 7.51±0.32 0.00±0.00 7.63±0.38 99.90±3.58
OS-46 2.55±0.16 28.87±2.30 0.00±0.00 85.08±2.16
OS-50 15.14±1.85 22.37±1.33 12.20±1.44 79.48±0.20
OS-66 16.53±0.52 24.02±0.44 39.13±0.75 93.71±3.55

表6

不同分类的各指标平均值的同等检验

指标Index F值F-value df1 df2 P值P-value
解有机磷能力Ability of organic phosphorus-solubilizing 0.432 3 70 0.730
解无机磷能力Ability of inorganic phosphate-solubilizing 0.805 3 70 0.495
解钾能力Ability of potassium-solubilizing 14.961 3 70 0.000
IAA含量IAA content 642.887 3 70 0.000

图2

燕麦内生真菌分类散点图

表7

判别分析函数特征值

判别函数
Discriminant function		 特征值
Eigenvalue		 方差贡献率
Variance contribution rate (%)		 累计贡献率
Accumulated contribution rate (%)		 典型相关系数
Canonical correlation coefficient
1 28.149 97.94 97.94 0.983
2 0.584 2.03 99.97 0.607
3 0.009 0.03 100.00 0.092
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