加拿大一枝黄花病原菌鉴定及其复配制剂的生防效果

doi:10.13560/j.cnki.biotech.bull.1985.2025-1218

摘要/Abstract

摘要：

目的获取能有效防治加拿大一枝黄花（Solidago canadensis L.）的病原菌，将其与化学除草剂进行复配，并评估复配制剂的盆栽防治效果。方法对染病的加拿大一枝黄花植株叶片和根部进行组织分离，筛选对加拿大一枝黄花具有致病性的真菌，通过形态学与分子生物学方法对其进行鉴定。在此基础上，筛选与该菌相容性良好的化学除草剂，研制复配可湿性粉剂，开展盆栽防效试验。结果分离得到的致病菌鉴定为大豆间座壳菌（Diaporthe sojae）SC02菌株。复配可湿性粉剂的配方为：3%吐温80，1%木质素磺酸钠，0.9%黄腐酸，0.5 μL/mL 20%氯氟吡氧乙酸，7%高岭土，40% SC02菌株孢子（1×10⁹ CFU/mL）。盆栽试验结果表明，在浓度为0.83-1.25 g/L时，该复配制剂对加拿大一枝黄花的株防效达70%-90%，鲜重防效达63%-73%，且未发现对周围其他植物产生不良影响。结论以SC02复配的可湿性粉剂质量符合国家要求，对加拿大一枝黄花株防效近90%，且复发率低，防除效果良好。

关键词: 加拿大一枝黄花, 大豆间座壳菌, 复配可湿性粉剂, 生物防治

Abstract:

Objective This study aimed to isolate pathogenic fungi with high efficacy against Canadian goldenrod (Solidago canadensis L.), develop a combined formulation integrating the fungal agent with a chemical herbicide, and evaluate its control efficacy in pot experiments. Method Pathogenic fungi from diseased S. canadensis leaves and roots were isolated, screened, and identified via morphological and molecular methods. Furthermore, chemical herbicides compatible with the fungi were screened, and a compound wettable powder was developed. The control efficacy of this formulation was then tested in pot experiments. Result The isolated pathogen was identified as Diaporthe sojae strain SC02. The compound wettable powder formulation consisted of the following components: 3% Tween 80, 1% sodium lignosulfonate, 0.9% fulvic acid, 0.5 μL/mL 20% fluroxypyr, 7% kaolin, and 40% SC02 strain spores (1×10⁹ CFU/mL). Pot experiment results showed that at concentrations ranging from 0.83 to 1.25 g/L, the compound formulation achieved a plant control efficacy of 70%–90% and a fresh weight control efficacy of 63%–73% against S. canadensis, with no adverse effects observed on surrounding plants. Conclusion The wettable powder formulated with SC02 meets national quality standards, exhibits nearly 90% control efficacy against S. canadensis plants, and has a low recurrence rate, demonstrating satisfactory weed control performance.

Key words: Solidago canadensis, Diaporthe sojae, combined wettable powder, biological control

靳文涛, 李迎新, 杨绍红, 刘应高, 翟森, 蒋平, 宋雷, 李姝江. 加拿大一枝黄花病原菌鉴定及其复配制剂的生防效果[J]. 生物技术通报, 2026, 42(10): 269-281.

JIN Wen-tao, LI Ying-xin, YANG Shao-hong, LIU Ying-gao, ZHAI Sen, JIANG Ping, SONG Lei, LI Shu-jiang. Pathogen Identification and Its Compound Formulations’ Control Efficacy against Solidago canadensis[J]. Biotechnology Bulletin, 2026, 42(10): 269-281.

图/表 13

表1 实时荧光定量PCR引物

Table 1 Primers for RT-qPCR

基因名称 Gene name	引物名称 Primer name	序列 Sequence （5′-3′）
ITS	ITS1	TCCCTAGCTGAACCTCCCC
ITS	ITS4	TCCTCCGCTTATTGATATCC
CAL	CAL-F	GAGTTCAAGGAGGCCTTCTCCC
CAL	CAL-R	CATCTTCTGGCCATCATGG
EF-1α	B.d-F	CATCGAGAAGTTCGAGAAGG
EF-1α	B.d-R	TACTTGAAGGAACCCTTACC
HIS	H3F	AGGTCCACTGGTGGCAAG
HIS	H3R	GCGGGCGAGCTGGATGTCCTT

表2 三种助剂最佳配比筛选

Table 2 Screening of the optimal ratio of three adjuvants

序号 Serial number	实验因素 Experimental factor
序号 Serial number	润湿剂 Wetting agent （%）	分散剂 Dispersant （%）	紫外保护剂 UV protectant （%）
1	1	1	0.5
2	5	3	0.5
3	3	5	0.5
4	5	1	0.7
5	3	3	0.7
6	1	5	0.9
7	3	1	0.9
8	1	3	0.9
9	5	5	0.9

表3 病原菌的分离结果

Table 3 Isolation results of pathogenic fungi

编号 No.	叶片分离株数 Number of leaf isolates	叶片分离率 Leaf separation rate （%）	根部分离株数 Number of root isolates	根部分离率 Root separation rate （%）
SC01	7	7	0	0
SC02	43	43	69	69
SC03	4	4	0	0
SC04	5	5	12	12
SC05	10	10	0	0
SC06	17	17	0	0
SC07	14	14	19	19

表4 筛选菌对加拿大一枝黄花及其非寄主植株的致病性

Table 4 Pathogenicity of screened strains against S. canadensis and its non-host plants

编号 No.	叶部病级 Leaf disease grade			根部病级 Root disease grade
编号 No.	加拿大一枝黄花 S. canadensis	香樟 C. camphora	楠木 P. zhennan	加拿大一枝黄花 S. canadensis	香樟 C. camphora	楠木 P. zhennan
SC01	1	2	1	0	0	0
SC02	5	1	0	5	0	0
SC03	1	0	1	0	0	0
SC04	2	2	2	0	0	1
SC05	2	3	3	0	0	0
SC06	1	4	5	0	0	0
SC07	4	5	4	1	0	0

图1 叶片与根部接种图A-C：叶片无菌琼脂块对照（香樟、楠木、加拿大一枝黄花，下同）；D-F：叶片接种SC02；G-I：叶片接种SC06；J-L：叶片接种SC07；M-O：根部接种SC02

Fig. 1 Leaf and root inoculation diagramA-C: Leaf sterile agar block control (C. camphora, P. zhennan, S. canadensis, the same below); D-F: leaf inoculation with SC02; G-I: leaf inoculation with SC06; J-L: leaf inoculation with SC07; M-O: root inoculation with SC02

图2 SC02菌株鉴定结果A：菌落图正面；B：菌落图背面；C：产孢培养基；D：子囊壳；E：子囊；F：α分生孢子；G：β分生孢子；H：菌株的系统发育进化树

Fig. 2 Identification results of SC02 strainA: Front view of the colony; B: back view of the colony; C: sporulation medium; D: ascus shell; E: ascus; F: α-conidia; G: β-conidia; H: phylogenetic tree of strains

图3 复配除草剂筛选结果A-H：单剂实验，分别为CK、20%氯氟吡氧乙酸、25%环嗪酮、41%草甘膦异丙铵盐、48%三氯吡氧乙酸丁氧基乙酯、10%精草胺膦胺盐、20%唏草酮、20%二甲四氯水剂组；I-P：化学除草剂推荐浓度筛选平板实验，分别为CK、20%氯氟吡氧乙酸、25%环嗪酮SL、41%草甘膦异丙铵盐、48%三氯吡氧乙酸丁氧基乙酯EC、10%精草胺膦胺盐、24%唏草酮、20%二甲四氯水剂组；Q-T：化学除草剂最适浓度筛选平板实验，20%氯氟吡氧乙酸、25%环嗪酮SL、41%草甘膦异丙铵盐、24%唏草酮组；U：加拿大一枝黄花复配除草剂筛选柱状图，图中推荐浓度为购买农药时官方推荐稀释浓度，最适浓度为推荐浓度稀释后病原菌最优生长浓度，不同小写字母表示同一时间不同处理间的差异显著性（P<0.001），下同

Fig. 3 Screening results of compound herbicidesA-H: Single-dose test. CK, 20% clopyralid, 25% carfentrazone, 41% glyphosate isopropylamine salt, 48% triclopyr butoxyethyl ester, 10% quizalofop-p-ethyl, 20% flumioxazin, 20% dimethachlor aqueous solution group, respectively. I-P: Screening plate for recommended concentrations of chemical herbicides. CK, 20% clopyralid, 25% carfentrazone SL, 41% glyphosate isopropylamine salt, 48% triclopyr butoxyethyl ester EC, 10% quizalofop-p-ethyl, 24% flumioxazin, 20% dimethachlor aqueous solution group, respectively. Q-T: Screening plate for optimal concentrations of chemical herbicides. 20% clopyralid, 25% carfentrazone SL, 41% glyphosate isopropylamine salt, 24% flumioxazin group, respectively. U: Bar chart of screening results for compound herbicides against S. canadensis. The recommended concentration refers to the dilution concentration officially recommended for the purchased herbicide. The optimal concentration indicates the dilution of the recommended concentration that supports the optimal growth of the pathogen. Different lowercase letters indicate significant differences among different treatments at the same time (P<0.001), the same below

表5 化学除草剂单剂筛选结果

Table 5 Screening results of individual chemical herbicide

化学除草剂 Chemical herbicides	推荐浓度 Recommended concentration	死亡率 Mortality rate （%）	死亡时间 Time of death （d）	死亡症状 Symptoms of death
20%氯氟吡氧乙酸	1 μL/mL	100	4	全株死亡，茎秆叶片全部萎蔫枯死
25%环嗪酮SL	0.02 g/mL	100	8	全株死亡，但有少量叶片茎秆泛绿，短时间未完全失活
41%草甘膦异丙铵盐	0.02 g/mL	100	6	全株死亡，但部分茎秆短时间未失活
48%三氯吡氧乙酸丁氧基乙酯EC	10 μL/mL	90	9	大部分植株枯死，但有少量植株茎秆存活
10%精草胺膦胺盐	10 μL/mL	100	8	全株死亡，茎秆叶片全部萎蔫枯死
24%唏草酮	1 μL/mL	100	5	全株死亡，茎秆叶片全部萎蔫枯死
20%二甲四氯水剂	5 μg/mL	100	4	全株死亡，茎秆叶片全部萎蔫枯死

图4 载体与3种助剂筛选结果A-E：载体（CK、硅藻土、高岭土、膨润土、硅胶）筛选结果；F-I：润湿剂（CK、吐温20、吐温80、PEG2000）筛选结果；J-Q：紫外保护剂（CK、紫外处理CK、黄腐酸、紫外处理黄腐酸、海藻酸钠、紫外处理海藻酸钠、糊精、紫外处理糊精）筛选结果；R-U：分散剂（CK、木质素磺酸钠、木质素磺酸钙、亚甲基双甲基萘磺酸钠）筛选结果； V-Y：载体、润湿剂、紫外保护剂、分散剂筛选结果

Fig. 4 Screening results for carriers and three adjuvantsA-E: Carrier (CK, diatomaceous earth, kaolin, bentonite, silica gel) screening results. F-I: Wetting agent (CK, Tween 20, Tween 80, PEG 2000) screening results. J-Q: Ultraviolet protection agent (CK, UV-treated CK, humic acid, UV-treated humic acid, sodium alginate, UV-treated sodium alginate, dextrin, UV-treated dextrin) screening results. R-U: Dispersant (CK, sodium lignosulfonate, calcium lignosulfonate, methylene bis-naphthalenesulfonate) screening results. V-Y: Screening results for carrier, wetting agent, ultraviolet protection agent, and dispersant

表6 不同润湿剂润湿时间测定

Table 6 Determination of wetting time of different wetting agents

润湿剂 Wetting agents	润湿时间 Wetting time （±SE）（s）
润湿剂 Wetting agents	1%	3%	5%
吐温20 Tween 20	32±0.58b	20±0.52b	11±1.88b
吐温80 Tween 80	24±0.68c	14±1.28c	10±0.75b
PEG2000	54±0.78a	45±1.05a	36±0.62a

图5 三种助剂最佳剂配比筛选结果

Fig. 5 Screening results of the optimal ratio of three additives

表7 大豆间座壳菌SC02可湿性粉剂复配剂质量检测结果

Table 7 Quality test results of wettable powder mixture of D. sojae SC02

指标 Index	国家标准值 National standards	测量值 Measured value
润湿时间 Wetting time （s）	≤180	36±1
悬浮率 Suspensibility （%）	≥70	74
含孢量 Spore content （CFU/g）	-	1.65×10⁹
pH	5.0-8.0	6.5
细度 Fineness （%）	≥90	92
起泡性 Frothiness （mL）	≤10 mL	7 mL
干燥减量 Drying reduction （%）	≤6	5.65
贮存稳定性 Storage stability （%）	≥80	≥81.5

图6 盆栽试验结果A-J：CK1清水、CK2氯氟吡氧乙酸，20、10、5、2.5、1.25、0.83、0.63、0.5 g/L稀释浓度处理效果；K-O：盆栽防治试验中株高、叶片枯死率、鲜重防效、根系枯死率、株防效结果

Fig. 6 Results of the pot experimentsA-J: CK1 distilled water, CK2 clopyralid, 20, 10, 5, 2.5, 1.25, 0.83, 0.63, 0.5 g/L dilution concentration treatment effect. K-O: Plant height, leaf necrosis rate, fresh weight control effect, root necrosis rate, and plant control effect results in potted control trials

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