杂草致病菌株GD-0221的分离、鉴定与除草潜力

doi:10.3969/j.issn.1004-1524.2022.09.15

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (9): 1967-1975.DOI: 10.3969/j.issn.1004-1524.2022.09.15

杂草致病菌株GD-0221的分离、鉴定与除草潜力

李祥(), 朱海霞()

青海大学农林科学院,农业农村部西宁作物有害生物科学观测实验站,青海省农业有害生物综合治理重点实验室,青海西宁 810016

收稿日期:2022-05-06 出版日期:2022-09-25 发布日期:2022-09-30
通讯作者: 朱海霞
作者简介:*朱海霞,E-mail: zhuhaixia0101163.com
李祥(1995—),男,甘肃武都人,硕士研究生,从事生物防治研究。E-mail: LX950301126.com
基金资助:
青海省中央引导地方科技发展资金(2022ZY034)

Isolation, identification and herbicidal potential of weed pathogenic strain GD-0221

LI Xiang(), ZHU Haixia()

Xining Crop Pest Scientific Observation and Experiment Station of the Ministry of Agriculture and Rural Affairs, Key Laboratory of Comprehensive Management of Agricultural Pests of Qinghai Province, Academy of Agricultural and Forestry Sciences, Qinghai University, Xining 810016, China

Received:2022-05-06 Online:2022-09-25 Published:2022-09-30
Contact: ZHU Haixia

摘要/Abstract

摘要：

为明确杂草致病菌株GD-0221的除草活性与作物安全性,利用组织分离法从患病甘草(Glycyrrhiza uralensis)根部分离获得杂草致病菌株GD-0221,通过形态学和分子生物学相结合确定该菌株的分类地位,利用菌饼接种法研究该菌株对5种阔叶杂草离体叶片的致病性,用发酵液喷雾接种法研究其对5种阔叶杂草的盆栽致病性和对青海主栽作物的安全性。根据菌株形态和DNA序列分析结果,将该菌株鉴定为尖孢镰刀菌(Fusarium oxysporum)。该菌株对5种阔叶杂草密花香薷(Elsholtzia densa)、藜(Chenopodium album)、冬葵(Malva verticillata)、酸模叶蓼(Polygonum lapathifolium)、萹蓄(Polygonum aviculare)均具有致病性,活体接种试验中杂草发病率为33.33%~100.00%,对密花香薷的致病效果最突出。作物安全性方面,该菌株对青稞(Hordeum vulgare)、小麦(Triticum aestivum)表现安全,而对油菜(Brassica campestr)、蚕豆(Vicia faba)和豌豆(Pisum sativum)表现不安全。GD-0221菌株具有开发成为禾本科作物田防除阔叶杂草微生物除草剂的潜力。

关键词: 尖孢镰刀菌, 除草活性, 作物安全性, 微生物除草剂

Abstract:

In order to clarify the herbicidal activity and crop safety of the weed pathogenic strain GD-0221, weed pathogenic strain GD-0221 was isolated from the root of diseased Glycyrrhiza uralensis by tissue isolation method. The taxonomic status of this strain was determined by morphological and molecular identification. The pathogenicity of the strain to the isolated leaves of five broadleaf weeds was determined by cake inoculation in vitro. The broth inoculation method was used to determine the pathogenicity to the five broadleaf weeds and the safety of the main crops in Qinghai in vivo. The strain was identified as Fusarium oxysporum by morphological and DNA sequence analysis, and it was pathogenic to five broadleaf weeds Elsholtzia densa, Chenopodium album, Malva verticillata, Polygonum lapathifolium and Polygonum aviculare. In vivo inoculation test result showed that pathogenicity rate of weeds was 33.33%-100.00%, it had the most prominent pathogenic effect on Elsholtzia densa. In terms of crop safety, the strain was safe to Hordeum vulgare, Triticum aestivum, but not safe to Brassica campestr, Vicia faba and Pisum sativum. Strain GD-0221 had the potential to be developed as a microbial herbicide against broadleaf weeds in gramineous crops.

Key words: Fusarium oxysporum, herbicidal activity, crop safety, microbial herbicide

中图分类号:

S476.1

李祥, 朱海霞. 杂草致病菌株GD-0221的分离、鉴定与除草潜力[J]. 浙江农业学报, 2022, 34(9): 1967-1975.

LI Xiang, ZHU Haixia. Isolation, identification and herbicidal potential of weed pathogenic strain GD-0221[J]. Acta Agriculturae Zhejiangensis, 2022, 34(9): 1967-1975.

图/表 7

图1 GD-0221菌株形态特征 A,GD-0221菌株正面形态;B,GD-0221菌株背面形态;C和D,GD-0221的分生孢子形态;E,GD-0221孢子梗形态。

Fig.1 Morphological characteristics of GD-0221 strain A, Positive morphology of GD-0221 strain; B, Back morphology of strain GD-0221; C and D, Conidia morphology of GD-0221; E, GD-0221 spore stalk morphology.

图2 基于GD-0221菌株5.8S-ITS rDNA基因序列构建的系统发育树

Fig.2 Phylogenetic tree based on 5.8S-ITS rDNA gene sequence of GD-0221 strain

图3 GD-0221菌株对5种阔叶杂草离体叶片致病性(接种7 d) A,密花香薷;B,藜;C,冬葵;D,酸模叶蓼;E,萹蓄。图4同。

Fig.3 Pathogenicity of strain GD-0221 to isolated leaves of five broadleaf weeds (7 days after inoculation) A, Elsholtzia densa; B, Chenopodium album; C, Malva verticillata; D, Polygonum lapathifolium; E, Polygonum aviculare. The same as in figure 4.

表1 菌株GD-0221对离体杂草的致病力比较(接种7 d)

Table 1 Comparison of pathogenicity of strain GD-0221 to weeds in vitro(7 days after inoculation)

杂草叶片 Weeds leaves	发病面积 Incidence area/cm²	发病等级 Incidence grade	发病特征 Incidence characteristics
密花香薷 E. densa	6.04	3	产生大块病斑,周围长出白色菌丝,叶片呈黑色枯死状 Large diseased spots were produced, white hyphae grow around, and the leaves were black and withered
藜 C. album	0.86	1	局部黄化,长有白色菌丝 Local yellowing with white hyphae
冬葵 M. verticillata	14.13	4	先出现病斑,之后黄化,最终叶片颜色变褐,表面有大量菌丝 Disease spots appeared first, then yellowing, and finally the leaf color turned brown, with a large number of hyphae on the surface
酸模叶蓼 P. lapathifolium	7.85	4	叶片完全枯死变褐,产生大量白色菌丝 The leaves completely withered and turned brown, producing a large number of white hyphae
萹蓄 P. aviculare	5.93	3	叶片表面产生病斑,之后逐渐扩展至整个叶片,表面有大量白色菌丝 Diseased spots appeared on the surface of the leaves, and then gradually extended to the whole leaf, with a large number of white hyphae on the surface

表2 菌株GD-0221对5种杂草的发病率与发病指数

Table 2 Pathogenicity and incidence index of strain GD-0221 to five kinds of weeds %

杂草 Weeds	发病率Incidence rate		病情指数Incidence index
杂草 Weeds	7 d	14 d	7 d	14 d
密花香薷E. densa	85.67±8.37 Aa	100.00 Aa	81.25±1.22 Aa	100.00 Aa
藜C. album	77.67±5.33 Aa	88.89±5.56 Aab	63.75±0.96 Cc	75.00±1.83 Bc
冬葵M. verticillata	20.00±11.55 Bb	33.33±6.67 Bc	13.75±0.21 Ee	27.50±0.39 De
酸模叶蓼P. lapathifolium	62.67±7.22 Aa	83.33±4.17 Ab	32.50±1.44 Dd	67.50±1.20 Cd
萹蓄P. aviculare	83.33±3.33 Aa	98.33±1.67 Aa	75.00±1.83 Bb	95.00±2.01 Ab

图4 GD-0221菌株对5种阔叶杂草的致病性(接种14 d)

Fig.4 Pathogenicity of strain GD-0221 to five broadleaf weeds (14 days after inoculation)

图5 GD-0221菌株对5种主栽作物安全性 A,青稞;B,小麦;C,豌豆;D,油菜;E,蚕豆。

Fig.5 Safety of strain GD-0221 on five main crops A, Hordeum vulgare; B, Triticum aestivum; C, Pisum sativum; D, Brassica campestris; E, Vicia faba.

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杂草致病菌株GD-0221的分离、鉴定与除草潜力

Isolation, identification and herbicidal potential of weed pathogenic strain GD-0221

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