枸杞内生真菌的筛选、鉴定及其生防作用

doi:10.3969/j.issn.1004-1524.20221161

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (7): 1662-1671.DOI: 10.3969/j.issn.1004-1524.20221161

枸杞内生真菌的筛选、鉴定及其生防作用

候彩霞¹^,²(), 丁德东¹^,², 何静¹^,²^,^*(), 赵吉桃¹^,², 李彦湘¹^,², 赵倩¹^,², 张崇庆¹^,², 李南¹^,²

1.甘肃农业大学林学院,甘肃兰州 730070
2.甘肃省枸杞无害化栽培工程研究中心,甘肃兰州 730070

收稿日期:2022-08-11 出版日期:2023-07-25 发布日期:2023-08-17
通讯作者: *何静,E-mail:hejing268@aliyun.com
基金资助:
甘肃省自然基金重点项目(20JR10RA508);国家自然科学基金(32060341)

Screening, identification and biocontrol effect of endophytic fungus from Lycium barbarum

HOU Caixia¹^,²(), DING Dedong¹^,², HE Jing¹^,²^,^*(), ZHAO Jitao¹^,², LI Yanxiang¹^,², ZHAO Qian¹^,², ZHANG Chongqing¹^,², LI Nan¹^,²

1. College of Forestry, Gansu Agricultural University, Lanzhou 730070, China
2. Gansu Lycium barbarum Harmless Cultivation Engineering Research Center, Lanzhou 730070, China

Received:2022-08-11 Online:2023-07-25 Published:2023-08-17
Contact: HE Jing

摘要/Abstract

摘要：

为筛选对枸杞根腐病主要致病菌Fusarium solani具有明显抑制效果的生防菌株,通过组织分离法从健康枸杞植株中分离内生真菌,用平板对峙法筛选拮抗菌株,结合形态特征和分子生物学鉴定明确其分类地位;分别采用菌丝生长速率法和悬滴法比较不同培养液处理的内生真菌对F. solani菌丝生长和孢子萌发的影响,通过盆栽试验验证其生防效果。结果表明,在分离得到的106株内生真菌中,筛选获得1株对F. Solani具有明显抑制效果的拮抗菌株,编号YBG8,经鉴定其为链格孢菌(Alternaria alternata);YBG8原液体积分数为20%时,对F. solani菌丝生长抑制率最强,达70.49%,且具有浓度依赖性;YBG8滤液体积分数50%时,对F. solani孢子萌发抑制率最大,达82.4%;盆栽试验结果显示,先接种A. alternata再接种F. solani,可有效降低枸杞根腐病的发病率,其相对防效可达50%。研究结果可以为有效利用内生真菌防治枸杞根腐病提供理论依据。

关键词: 枸杞, 根腐病, 内生真菌, 拮抗菌株, 发病率

Abstract:

In order to screen antagonistic strains with significant inhibitory effects against Fusarium solani, the main pathogenic fungus of Lycium barbarum root rot, endophytic fungi were isolated from healthy L. barbarum plants using tissue separation method. Antagonistic strains were selected through plate confrontation method, and their taxonomic status was determined based on morphological features and molecular biology identification. The effects of endophytic fungi treated with different culture media on the mycelial growth and spore germination of F. solani were compared by mycelial growth rate method and droplet method, and their biocontrol effects were verified by pot experiments. The results showed that out of the 106 endophytic fungi isolated, one antagonistic strain YBG8 was selected, which had a significant inhibitory effect against F. solani, and was identified as Alternaria alternata. The inhibition rate of YBG8 against F. solani mycelial growth was the highest (70.49%) at a volume fraction of 20%, and showed concentration dependence. When YBG8 filtrate volume fraction was 50%, the inhibition rate against F. solani spore germination was the highest (82.4%). Pot experiments showed that pre-inoculation with A. alternata followed by F. solani could effectively reduce the incidence of L. barbarum root rot, achieving a relative preventive effect of up to 50%. These findings provided a theoretical basis for the effective use of endophytic fungi to control L. barbarum root rot.

Key words: Lycium barbarum, root rot, endophytic fungus, antagonistic strain, disease incidence

中图分类号:

S182

候彩霞, 丁德东, 何静, 赵吉桃, 李彦湘, 赵倩, 张崇庆, 李南. 枸杞内生真菌的筛选、鉴定及其生防作用[J]. 浙江农业学报, 2023, 35(7): 1662-1671.

HOU Caixia, DING Dedong, HE Jing, ZHAO Jitao, LI Yanxiang, ZHAO Qian, ZHANG Chongqing, LI Nan. Screening, identification and biocontrol effect of endophytic fungus from Lycium barbarum[J]. Acta Agriculturae Zhejiangensis, 2023, 35(7): 1662-1671.

图/表 8

图1 五株内生真菌对F. solani的抑制作用

Fig.1 Inhibition of F. solani by 5 endophytic fungi

图2 五株拮抗菌株对F. solani的抑菌率数据为平均值±标准误,柱上无相同小写字母表示差异显著(P<0.05)。

Fig.2 Inhibition rate of 5 antagonistic strains to F. solani Data ($\bar{x}$±s) on the bars marked without the same lowercase letter indicated significant differences at P<0.05.

图3 YBG8菌株的形态特征 A,培养3 d YBG8菌落形态正面;B,培养3 d YBG8菌落形态背面;C,培养7 d YBG8菌落形态正面;D,培养7 d YBG8菌落形态背面;E、F分别为YBG8菌丝和孢子形态。

Fig.3 Morphological characteristics of YBG8 strain A, The front of YBG8 colony morphology cultured for 3 days; B, The back of YBG8 colony morphology cultured for 3 days; C, The front of YBG8 colony morphology cultured for 7 days; D, The back of YBG8 colony morphology cultured for 7 days; E and F, Mycelia and spore morphology of YBG8.

图4 YBG8与其他菌株基于rDNA-ITS序列的系统发育树

Fig.4 Phylogenetic tree of YBG8 and other strains based on rDNA-ITS sequences

表1 YBG8菌株培养原液、滤液和失活液对F. solani菌丝的抑制率

Table 1 Inhibition rate of culture solution, filtrate solution and inactivated solution of YBG8 on F. Solani mycelia %

体积分数 Volume fraction/%	对F. solani的抑制率 Inhibition rate on F. Solani mycelia
体积分数 Volume fraction/%	培养原液 Culture solution	培养滤液 Filtrate solution	失活液 Inactivated solution
1.25	31.40±0.44	0.91±0.54	0.13±0.01
2.50	34.54±0.28	3.01±0.54	0.12±0.01
5	37.04±0.58	4.34±0.61	0.11±0.01
10	49.54±0.17	8.22±0.85	0.10±0.01
20	70.49±0.63	18.75±0.73	0.10±0.01

图5 菌株YBG8培养滤液对F. solani孢子萌发的影响

Fig.5 Effect of fermentation filtrate of strain YBG8 on spore germination of F. solani

图6 YBG8菌株对枸杞根腐病的防效 A,F. solani;B,对照;C,YBG8;D,接种YBG8菌株 7 d后接种F. solani。

Fig.6 Control effect of YBG8 strain on L.barbarum root rot A, F. solani; B, Control; C, YBG8; D, After 7 days of inoculation with YBG8 strain, F. solani was inoculated.

表2 菌株YBG8培养液对枸杞根腐病的防治效果

Table 2 Control effect of culture solution of YBG8 strain on wolfberry root rot %

处理 Treatment	发病率 Morbidity	病情指数 Disease index	相对防效 Relative control effect
CK	50.0	40.0
YBG8	40.0	36.0	38.8
YBG8+F. solani	30.0	30.0	50.0
F. solani	70.0	60.0	-40
精甲·咯·嘧菌	20	20	66.7
Metalaxyl-M·fludioxonil· azoxystrobin

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枸杞内生真菌的筛选、鉴定及其生防作用

Screening, identification and biocontrol effect of endophytic fungus from Lycium barbarum

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