一株茶根腐病拮抗真菌的筛选鉴定与生物学特性研究

doi:10.3969/j.issn.1004-1524.20240565

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (7): 1469-1480.DOI: 10.3969/j.issn.1004-1524.20240565

一株茶根腐病拮抗真菌的筛选鉴定与生物学特性研究

邓玉莲¹^,²(), 谭琳³, 牛丽¹^,², 卢前程¹^,², 周玲红⁴, 李桂花¹^,², 胡秋龙¹^,²^,^*()

1.湖南农业大学茶学教育部重点实验室,湖南长沙 410128
2.湖南农业大学园艺学院,湖南长沙 410128
3.湖南农业大学植物保护学院,湖南长沙 410128
4.郴州市农业科学研究所,湖南郴州 423001

收稿日期:2024-06-27 出版日期:2025-07-25 发布日期:2025-08-20
作者简介:邓玉莲(2000—),女,湖南岳阳人,硕士研究生,研究方向为茶树病虫害。E-mail:2214194556@qq.com
通讯作者: ^*胡秋龙,E-mail:huqiulongnet@126.com
基金资助:
国家重点研发计划(2022YFD16000803-6);湖南省重点研发计划(2022NK2051);湖南省科技创新重大项目(2021NK1020)

Screening and identification of an antagonistic fungus strain against root rot disease in tea plants and its biological characteristics

DENG Yulian¹^,²(), TAN Lin³, NIU Li¹^,², LU Qiancheng¹^,², ZHOU Linghong⁴, LI Guihua¹^,², HU Qiulong¹^,²^,^*()

1. Key Lab of Tea Science, Ministry of Education, Hunan Agricultural University, Changsha 410128, China
2. College of Horticulture, Hunan Agricultural University, Changsha 410128, China
3. College of Plant Protection, Hunan Agricultural University, Changsha 410128, China
4. Chenzhou Institute of Agricultural Science, Chenzhou 423001, Hunan, China

Received:2024-06-27 Online:2025-07-25 Published:2025-08-20

摘要/Abstract

摘要： 采用平板对峙法,从茶树根际土壤中筛选出一株对茶根腐病病原菌具有拮抗效果的真菌,并利用形态学和分子生物学方法对其进行鉴定,研究其生物学特性,测定其抑菌谱、代谢物质抑菌能力和产胞外水解酶的活性。结果表明,分离筛选出的菌株FX2为橘青霉(Penicillium citrinum),对茶树根腐病病原菌Fusarium cugenangense具有较好的拮抗作用,抑菌率为72.98%。菌株FX2对碳氮比的响应不敏感,最适碳源是乳糖,最适氮源是蛋白胨或酵母提取物,最适硫酸盐为MgSO₄或K₂SO₄,对NaCl的耐受范围为0~140 g·L^-1,最适pH值为6,最高耐受温度为60 ℃,紫外光照射会对该菌株的生长产生一定的抑制作用。该菌株对链格孢菌(Alternaria alternata)、禾谷镰孢菌(Fusarium graminearum)、大斑病凸脐蠕孢(Exserohilum turcicum)、柑橘间座壳菌(Diaporthe citri)、尖孢镰刀菌(Fusarium oxysporum)、葱刺盘孢(Colletotrichum circinans)、茶炭疽病菌刺盘孢菌(Colletotrichum camelliae)、茶拟盘多毛孢(Pseudopestalotiopsis theae)8种植物病原菌的抑制率均在50%以上,具有较好的抗菌广谱性。该菌株5%、10%、15%、20%的无菌发酵滤液对茶树根腐病病原菌的抑制率分别为35.12%、45.98%、52.80%、59.46%,非挥发性代谢物、挥发性代谢物对病原菌的抑制率分别为71.73%、22.14%。此外,该菌株还具有分泌蛋白酶、淀粉酶、几丁质酶和纤维素酶的能力。综上,分离得到的菌株FX2是具有较好前景的茶树根腐病病原菌生防资源。

关键词: 茶树, 根腐病, 拮抗真菌, 抑制率, 生物学特性

Abstract:

In the present study, a fungal isolate with antagonistic effect on tea root rot pathogen was screened from the rhizosphere soil of tea plants infected by tea root rot pathogen through plate confrontation method. The isolated strain was identified by the morphological and molecular biology methods, and its biological characteristics were studied, as well as its antimicrobial spectrum, antimicrobial ability of metabolites and extracellular hydrolase activities. The results showed that the obtained strain FX2 was finally identified as Penicillium citrinum, which exhibited an inhibition rate of 72.98% against the pathogen Fusarium cugenangense of tea root rot. The strain FX2 was relatively insensitive to the carbon-nitrogen ratio. The suitable carbon source of this strain was lactose, the suitable nitrogen source was peptone or yeast extract, the optimal sulfate was MgSO₄ or K₂SO₄. The tolerance range of this strain to NaCl was 0-140 g·L^-1, the optimal pH value was 6, and the maximum withstand temperature was 60 ℃. The ultraviolet light irradiation exhibited certain inhibitory effect on the growth of this strain. The inhibition rate of this strain against other 8 plant pathogens (including Alternaria alternata, Fusarium graminearum, Exserohilum turcicum, Diaporthe citri, Fusarium oxysporum, Colletotrichum circinans, Colletotrichum camelliae, Pseudopestalotiopsis theae) were higher than 50%, indicating broad antibacterial spectrum. The inhibition rate of 5%, 10%, 15% and 20% sterile fermentation filtrate against root rot pathogen was 35.12%, 45.98%, 52.80% and 59.46%, respectively, and the inhibition rate of non-volatile metabolites and volatile metabolites against root rot pathogen was 71.73% and 22.14%, respectively. Besides, this strain also had the ability to secrete proteases, amylases, chitinases and cellulases. In conclusion, the isolated strain FX2 was a promising biocontrol resource for tea root rot.

Key words: tea tree, root rot, antagonistic fungi, inhibition rate, biological properties

中图分类号:

S432

邓玉莲, 谭琳, 牛丽, 卢前程, 周玲红, 李桂花, 胡秋龙. 一株茶根腐病拮抗真菌的筛选鉴定与生物学特性研究[J]. 浙江农业学报, 2025, 37(7): 1469-1480.

DENG Yulian, TAN Lin, NIU Li, LU Qiancheng, ZHOU Linghong, LI Guihua, HU Qiulong. Screening and identification of an antagonistic fungus strain against root rot disease in tea plants and its biological characteristics[J]. Acta Agriculturae Zhejiangensis, 2025, 37(7): 1469-1480.

图/表 10

图1 菌株FX2对茶树根腐病病原菌的抑制作用 A,对照组(CK);B,添加有FX2菌株的处理组。

Fig.1 Inhibitory effect of strain FX2 against Fusarium cugenangense A, Control (CK); B, Treatment with strain FX2.

图2 菌株FX2对茶树根腐病病原菌菌丝生长的影响 A,对照组(CK);B,添加有FX2菌株的处理组。

Fig.2 Effect of strain FX2 on hypha growth of Fusarium cugenangense A, Control (CK); B, Treatment with strain FX2.

图3 菌株FX2的菌落形态与显微特征 A和B、C和D、E和F分别为菌株FX2在OA、MEA、PDA培养基平板上28 ℃培养3 d的正反面形态。G、H分别展示菌株FX2在PDA培养基平板上的菌丝、孢子形态。

Fig.3 Colony and morphological characteristics of strain FX2 A and B, C and D, E and F show the front and back of colony morphologies of strain FX2 cultured on OA、MEA、PDA medium at 28 ℃ for 3 days, respectively. G and H show the morphology of mycelium and spores of strain FX2 cultured on PDA medium, respectively.

图4 基于菌株 FX2 ITS 序列构建的系统进化树分支处数值表示bootstrap值。标尺0.01代表序列的进化差异。带T的为标准菌株。

Fig.4 Phylogenetic tree of strain FX2 based on ITS sequence Numbers at branch nodes present bootstrap value. The scale bar 0.01 represents sequence variance. “T” indicates standard strain.

图5 菌株FX2的生物学特性 CK表示对照。柱上无相同字母的表示差异显著(P<0.05)。下同。

Fig.5 Biological characteristics of strain FX2 CK represents the control treatment. Bars marked without the same letters indicate significant difference at P<0.05. The same as below.

图6 菌株FX2对8种植物病原菌的抑制作用

Fig.6 Inhibition effect of strain FX2 against 8 plant pathogens

表1 菌株FX2无菌发酵滤液的抑菌能力

Table 1 Inhibition effect of sterile fermentation filtrate of strain FX2

无菌发酵滤液的体积分数 Volume fraction of sterile fermentation filtrate/%	菌落直径 Colony diameter/mm	抑制率 Inhibition rate/%
0	78.00±0.41 a	—
5	50.60±0.43 b	35.12±0.56 d
10	42.13±0.81 c	45.98±1.04 c
15	36.80±1.16 d	52.80±1.49 b
20	31.61±0.45 e	59.46±0.57 a

图7 菌株FX2挥发性代谢物的抑菌活性左侧为对照组,右侧为处理组。图8同。

Fig.7 Inhibition effect of volatile metabolites of strain FX2 Left and right show control and treatment, respecitvley. The same as in Fig. 8.

图8 菌株FX2难挥发性代谢物的抑菌活性

Fig.8 Inhibition effect of non-volatile metabolites of strain FX2

图9 菌株FX2产胞外水解酶的能力 A,蛋白酶;B,淀粉酶;C,几丁质酶;D,纤维素酶。

Fig.9 Abilities of strain FX2 to produce extracellular hydrolase A, Protease; B, Amylase; C, Chitinase; D, Cellulase。

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一株茶根腐病拮抗真菌的筛选鉴定与生物学特性研究

Screening and identification of an antagonistic fungus strain against root rot disease in tea plants and its biological characteristics

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