老枞根际放线菌鉴定及其对可可毛色二孢抑制效果研究

陈小尘; 徐金荣; 高培妍; 罗正朵; 郑智胜; 吴宝川; 周艳; 黄兆斌; 陈洪彬; 张秋芳

doi:10.19303/j.issn.1008-0384.2023.09.012

老枞根际放线菌鉴定及其对可可毛色二孢抑制效果研究

doi: 10.19303/j.issn.1008-0384.2023.09.012

1.
泉州师范学院海洋与食品学院，福建　泉州　362000
2.
福建农林大学食品科学学院，福建　福州　350002
3.
武夷山国家公园科研监测中心，福建　南平　354300

基金项目: 福建省科技计划引导性项目(2020N0032)

详细信息

作者简介:
陈小尘（1997 —），男，硕士研究生，主要从事放线菌抗菌活性物质提取研究，E-mail：872771920@qq.com

通讯作者:
张秋芳（1973 —），女，博士，教授，主要从事环境微生物生态、生物资源利用和生态修复研究，E-mail: qfzhang@163.com

中图分类号: TS201.3
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出版历程
- 收稿日期: 2023-04-17
- 修回日期: 2023-06-13
- 网络出版日期: 2023-10-13
- 刊出日期: 2023-09-28

Identification and Inhibitory Effect on Lasiodiplodia theobromae of Actinomycetes in Camellia sinensis Rhizosphere Soil

1.
College of Oceanology and Food Science, Quanzhou Normal University, Quanzhou, Fujian　362000, China
2.
College of Food Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350007, China
3.
Research Monitoring Center, Wuyishan National Park, Nanping, Fujian　354300, China

摘要

摘要: 目的从老枞根际土壤分离可培养放线菌，获取潜在新物种，并筛选可可毛色二孢菌拮抗菌株。方法采用土壤稀释涂布法和三区划线法进行分离和纯化，获得放线菌菌株；通过比对16S rRNA基因序列和构建系统发育树，对菌株进行初步分类和鉴定；采用平板对峙法进行抑菌活性筛选。结果（1）共获得81株不同放线菌菌株，分别隶属于链霉菌Streptomyces（54.32%）、节杆菌Arthrobacter（27.16%）、微杆菌Microbacterium（11.11%）、北里孢菌Kitasatospor（4.94%）和短小杆菌Curtobacterium（2.47%）5个属；（2）与已知模式菌株相比，相似度小于98.65%的潜在新种共有20 株；（3）经可可毛色二孢平板对峙试验筛选后，获得7株高抑菌活性链霉菌，其中链霉菌ON316885的抑菌率最高，达到63.92%。结论武夷山老枞根际土壤中蕴含着丰富的放线菌资源，具有深度挖掘研究的价值，其中一些链霉菌具有较高的生防应用价值。
- 老枞根际土壤 /
- 放线菌 /
- 可可毛色二孢 /
- 16S rRNA基因
Abstract: Objective Actinomycetes collected from the rhizosphere soils of old tea plants were isolated to screen for species unknown previously and their potential pathogenic inhibitory effect. Method Soil specimens were gathered from the target lots at Shuixian, Fujian to isolate actinomycetes by plate culture. Phylogenetic trees based on 16S rRNA gene for the isolates were constructed and diversity index calculated in the tea plant rhizosphere soils. Result (1) Eighty-one actinomycete strains were obtained that belonged to Streptomyces(54.32%), Arthrobacter (27.16%), Microbacterium (11.11%), Kitasatospora(4.94%), and Curtobacterium (2.47%). (2) In comparing with the reference strain, 20 of the 81 taxa showed less than 98.65% similarity. (3) Seven strains of the Streptomyces significantly inhibited the growth of Lasiodiplodia theobromae, of which, the one code-named WYS-24 reached the highest rate at 63.92%. Conclusion Abundantly diverse actinomycetes resided in the rhizosphere of old tea plants in Wuyishan region. They were a rich resource for in-depth studies and development for biological inhibitors against plant pathogens.
- Rhizosphere /
- Camellia sinensis /
- Lasiodiplodia theobromae /
- 16S rRNA gene

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图 1 武夷山老枞茶园根际土壤采样点分布

Figure 1. Sites for sampling actinomycetes in old tea plant rhizosphere soils at plantations in Wuyishan

下载: 全尺寸图片幻灯片

图 2 放线菌物种组成

A. 放线菌属水平物种组成；B. 链霉菌组成；C. 节杆菌组成。

Figure 2. Actinomycetes taxa

A: Genera of Actinomycetes; B: species of Streptomyces; C: species of Arthrobacter.

下载: 全尺寸图片幻灯片

图 3 老枞根际土壤中可培养放线菌系统发育进化树

粗体表示与模式菌株相似度小于98.65%。

Figure 3. Phylogenetic tree of culturable actinomycetes in old tea plant rhizosphere soil

Bold font indicates less than 98.65% similarity with reference strain.

下载: 全尺寸图片幻灯片

图 4 强拮抗活性菌株对可可毛色二孢生长的抑制效果

A. 平板正面；B. 平板背面。

Figure 4. Inhibitory effects of highly active actinomycetes against L. theobromae

A: Front of petri dish; B: back of petri dish.

下载: 全尺寸图片幻灯片

表 1 老枞根际土壤可培养放线菌代表菌株与最相似模式菌株比较

Table 1. Comparison between actinomycetes isolated from old tea plant rhizosphere and that closest to reference strain

菌株登录号 Accession No. of strain	最相似菌株及登录号 The closest type strain and its accession	中文名称 Chinese name	相似度 Similarity/%
ON165519	Microbacterium proteolyticum (KM359785)	蛋白水解微杆菌	98.08
ON165520	Microbacterium testaceum (BJML01000022)	种皮微杆菌	98.53
ON165521	Streptomyces acidiscabies (D63865)	酸性疮链霉菌	97.91
ON165522	Streptomyces pseudovenezuelae (KQ948163)	金霉素链霉菌	97.56
ON165523	Streptomyces acidiscabies (D63865)	酸性疮链霉菌	97.94
ON165524	Arthrobacter livingstonensis (GQ406811)	利文斯顿岛节杆菌	97.97
ON165525	Arthrobacter bambusae (KF150696)	竹节杆菌	98.63
ON165526	Arthrobacter bambusae (KF150696)	竹节杆菌	98.42
ON165527	Streptomyces xiamenensis (EF012099)	厦门链霉菌	97.55
ON165528	Streptomyces alanosinicus (AB184442)	阿拉诺链霉菌	97.43
ON165529	Streptomyces graminifolii (HQ267984)	禾叶链霉菌	98.58
ON165530	Streptomyces acidiscabies (D63865)	酸性疮链霉菌	97.96
ON165531	Arthrobacter livingstonensis (GQ406811)	利文斯顿岛节杆菌	97.46
ON165532	Streptomyces acidiscabies (D63865)	酸性疮链霉菌	98.56
ON165533	Streptomyces philanthi (DQ375802)	嗜酸链霉菌	96.73
ON165534	Kitasatospora psammotica (MT760550)	—	98.54
ON165535	Streptomyces intermedius (AB184277)	中间型链霉菌	98.61
ON165536	Microbacterium proteolyticum (KM359785)	蛋白水解微杆菌	98.22
ON165537	Streptomyces griseocarneus (MT760576)	灰色链霉菌	97.80
ON165538	Streptomyces aquilus (MH718844)	棕色链霉菌	97.78
ON316876	Streptomyces ardesiacus (DQ026631)	桔橙链霉菌	99.16
ON316877	Streptomyces pratensis (JQ806215)	草地链霉菌	99.56
ON316878	Streptomyces mexicanus (AF441168)	墨西哥链霉菌	98.92
ON316879	Streptomyces halstedii (AB184142)	郝氏链霉菌	99.86
ON316880	Streptomyces microflavus (AB184284)	细黄链霉菌	99.36
ON316881	Streptomyces badius (AY999783)	栗褐链霉菌	99.03
ON316882	Streptomyces zaomyceticus (AB184346)	沙阿霉素链霉菌	98.95
ON316883	Streptomyces niveus (DQ442532)	雪百链霉菌	100.00
ON316884	Streptomyces xylanilyticus (LC128341)	—	99.92
ON316885	Streptomyces rochei (MUMD01000370)	罗氏链霉菌	98.88
ON316886	Streptomyces coelicoflavus (AB184650)	天蓝黄链霉菌	98.95
ON316887	Kitasatospora xanthocidica (AB184427)	—	99.50
ON316888	Arthrobacter bambusae (KF150696)	竹节杆菌	99.50
ON316889	Arthrobacter gyeryongensis (JX141781)	绞股蓝节杆菌	99.06
ON316890	Microbacterium proteolyticum (KM359785)	—	99.02
ON316891	Microbacterium azadirachtae (JYIT01000023)	—	99.29
ON316892	Microbacterium paraoxydans (BCRH01000180)	—	99.22
ON316893	Curtobacterium citreum (X77436)	柠檬色短小杆菌	99.36
ON316894	Curtobacterium albidum (AM042692)	—	98.79
粗体表示与模式菌株的相似度小于98.65%。 Bold font indicates a less than 98.65% similarity with reference strain.

下载: 导出CSV

表 2 强拮抗活性菌株对可可毛色二孢的抑制作用

Table 2. Inhibitory effects of highly active actinomycetes against L. theobromae

菌株登记号 Accessions of strain	病原菌直径 Diameter of pathogen/mm	抑制率 Inhibition rate/%
CK	90.000	—
ON316884	59.725±0.111	33.64±0.37
ON165521	54.923±0.086	38.97±0.25
ON165523	52.211±0.095	41.99±0.25
ON165532	34.106±0.108	62.10±0.19
ON316885	32.476±0.106	63.92±0.18
ON165522	35.657±0.097	60.38±0.18
ON165537	39.736±0.062	55.85±0.12

下载: 导出CSV

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