福建省芋疫霉交配型测定及防治药剂筛选

王荣波; 石茗月; 徐月华; 刘裴清; 卞润恬; 李本金; 翁启勇

福建省芋疫霉交配型测定及防治药剂筛选

王荣波^{1, 2,},
石茗月^{1, 2},
徐月华³,
刘裴清^{1, 2},
卞润恬^{1, 2},
李本金^{1, 2, ,},
翁启勇^{1, 2, ,}

1.
福建省农业科学院植物保护研究所/福建省作物有害生物监测与治理重点实验室/福建省作物有害生物绿色防控工程研究中心，福建　福州　350013
2.
福建农林大学植物保护学院/海峡两岸特色作物安全生产省部共建协同创新中心，福建　福州　350002
3.
福建省龙岩市永定区植保植检站，福建　龙岩　364100

基金项目: 国家自然科学基金（32072400）；福建省农业科学院科技创新团队建设项目（CXTD2021002-1）；厦门市重大科技项目（3502Z20221018）；福建省农业高质量发展超越“5511”协同创新工程项目（XTCXGC2021011、 XTCXGC2021017）

详细信息

作者简介:
王荣波（1988 —），男，博士，助理研究员，主要从事植物病理及分子生物学研究，E-mail：wrb16128@163.com

通讯作者:
李本金（1969 —），女，研究员，主要从事植物病理及分子生物学研究，E-mail：lbenjin@126.com

翁启勇（1962 — ），男，硕士，研究员，主要从事作物有害生物综合治理技术研究，E-mail: wengqy@faas.cn

中图分类号: S436.67
计量
- 文章访问数: 106
- HTML全文浏览量: 39
- PDF下载量: 27
- 被引次数: 0
出版历程
- 收稿日期: 2024-04-02
- 修回日期: 2024-05-13
- 网络出版日期: 2024-08-15

Mating Type and Effective Control of Phytophthora colocasiae

WANG Rongbo^{1, 2
,},
SHI Mingyue^{1, 2},
XU Yuehua³,
LIU Peiqing^{1, 2},
BIAN Runtian^{1, 2},
LI Benjin^{1, 2
, ,},
WENG QiYong^{1, 2
, ,}

1.
Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests, Fujian Engineering Research Center for Green Pest Management, Institute of Plant Protection, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350013, China
2.
Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops, College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
3.
Yongding Plant Protection and Inspection Station of Longyan City, Longyan, Fujian　364100, China

摘要

摘要: 目的明确福建省芋疫霉交配型，筛选可用于防治芋疫病的化学药剂。方法对2020年采集自福建4个地区的芋疫病样本进行病原菌进行分离纯化，结合形态特征观察、致病性测定和Ypt1序列同源性分析对病原菌进行鉴定，通过与辣椒疫霉A1、A2交配型菌株对峙培养来测定其交配型，利用菌丝生长法测定6种杀菌剂对病原菌的室内毒力。结果通过形态特征观察、致病性测定和Ypt1序列同源性分析，明确所分离病原菌均为芋疫霉（Phytophthora colocasiae）。交配型测定发现，所分离的125个芋疫霉菌株中，122个为A2交配型，3个为A1A2交配型；室内药剂筛选结果表明，98%甲霜灵对芋疫霉抑菌效果最好，EC₅₀值为0.146±0.032 μg·mL⁻¹；95%烯酰吗啉、98%氟吡菌胺和94%氰霜唑抑菌效果较好，EC₅₀值介于（0.239±0.011）~（0.713±0.088） μg·mL⁻¹；而95%嘧菌酯的抑菌活性最低，EC₅₀值为23.447±3.666 μg·mL⁻¹。结论福建省芋疫霉以A2交配型为优势群体，甲霜灵、烯酰吗啉、氟吡菌胺和氰霜唑可作为芋疫病高效防控的轮换使用药剂。
- 芋疫霉 /
- 病原菌鉴定 /
- 交配型 /
- 防治药剂
Abstract: Objective Mating type and fungicides for effective control of Phytophthora colocasiae that caused 2020 taro blight epidemic in Fujian were determined. Method Specimens of diseased taro tissues were collect from the 4 blight-infected regions in Fujian to isolate and identify the pathogen. Based on the morphology, pathogenicity, and sequence homology of Ypt1, the pathogenic strains were identified to be of P. colocasiae. Subsequently, mating type and sensitivity to 6 fungicides of the isolates were determined in the laboratory. Result In total, 125 strains were isolated and identified to have caused the epidemic. Out of them, 122 belonged to the A2 mating type and 3 the A1A2 type. The laboratory toxicity test of 6 fungicides on the isolates showed 98% metalaxyl to be the strongest with EC₅₀ of 0.146±0.032 μg·mL⁻¹, while the EC₅₀ of 95% dimethomorph, 98% fluopicolide, and 94% cyazofamid ranged from 0.239±0.011 μg·mL⁻¹ to 0.713±0.088 μg·mL⁻¹ and that of 95% azoxystrobin at 23.447±3.666 μg·mL⁻¹. Conclusion The dominant strains of P. colocasiae that caused the taro blight in Fujian in 2020 were of the A2 mating type and could be best controlled by using 98% metalaxyl, 95% dimethomorph, 98% fluopicolide, or 94% cyazofamid.
- Phytophthora colocasiae /
- pathogen identification /
- mating type /
- fungicides

HTML全文

图 1 芋疫病的田间发病症状

A～B：芋疫病叶片症状；C：芋疫病茎秆发病症状；D：芋疫病球茎发病症状。

Figure 1. Symptoms of taro blight in the field

A-B: Symptom on infected leaves; C: symptom on infected stems; D: symptom on infected tubers.

下载: 全尺寸图片幻灯片

图 2 病原菌的致病性测定

接种5 d后芋叶片正面（A）和背面（B）发病症状，a～c: 来自福建不同地区的代表性分离菌株。

Figure 2. Pathogenicity of isolates

Disease symptoms on front (A) and back (B) of taro leaves 5 d after inoculation; a-c: representative isolates from various districts in Fujian.

下载: 全尺寸图片幻灯片

图 3 病原菌的形态特征

A：菌落形态；B：菌丝；C：孢子囊。

Figure 3. Morphology of blight pathogen

A: Colonies of isolated strains; B: mycelia; C: sporangium.

下载: 全尺寸图片幻灯片

图 4 基于ITS-LSU-Ypt1基因联合序列构建的系统发育树

每个分支上方的数值表示来自邻接法的自展值，以1000次重复的百分比表示；红色菱形代表分离的病原菌株。

Figure 4. Phylogenetic tree of FZPc-1, LYPc-1, NDPc-1, and NPPc-1 based on ITS-LSU-Ypt1 sequences by NJ method

Data above each branch indicate bootstrap values from neighbor-joining method as percentages in 1,000 replicates; red diamonds indicate isolated pathogens.

下载: 全尺寸图片幻灯片

图 5 福建省芋疫霉菌株交配型测定

A: 菌株单独培养；B：菌株与辣椒疫霉A1交配型菌株对峙培养；C：菌株与辣椒疫霉A2交配型菌株对峙培养；D：藏卵器、卵孢子和雄器。

Figure 5. Mating types of P. colocasiae from Fujian

A: Culture of individual P. colocasiae isolate; B: confrontation culture between isolate and P. capsici strain of A1 mating type; C: confrontation culture between isolate and P. capsici strain of A2 mating type; D: oogonium, oospore, and antheridium.

下载: 全尺寸图片幻灯片

表 1 福建不同地区芋疫霉交配型构成及地理分布

Table 1. Mating types and distribution of P. colocasiae strains in Fujian

地区 Location	菌株数 Number of isolates	A2交配型 A2 mating type		A1A2交配型 A1A2 mating type
地区 Location	菌株数 Number of isolates	菌株数 Number of isolates	频率 Frequency /%	菌株数 Number of isolates	频率 Frequency/%
龙岩 Longyan	37	35	28.0	2	1.6
福州 Fuzhou	31	31	24.8	0	0
南平 Nanping	29	28	22.4	1	0.8
宁德 Ningde	28	28	22.4	0	0
总计 Total	125	122	97.6	3	2.4

下载: 导出CSV

表 2 6种杀菌剂对福建省不同地区芋疫霉的室内毒力测定结果

Table 2. Inhibition effects of 6 fungicides against P. colocasiae from regions infected by taro blight in Fujian

杀菌剂 Fungicides	菌株来源 Source of isolate	毒力回归方程 Toxicity equation	相关系数 Correlation coefficient(r)	EC₅₀ /(μg·mL⁻¹)	均值±标准误 Mean±SE /(μg·mL⁻¹)
98% 甲霜灵 98% Metalaxyl	龙岩	y=1.2251x + 7.7317	0.9991	0.108	0.146±0.032
	福州	y=1.404x + 7.3045	0.9790	0.194
	南平	y=0.9833x + 7.0201	0.9871	0.128
	宁德	y=0.9689x + 6.81	0.9780	0.154
98% 霜脲氰 98% Cymoxanil	龙岩	y=0.3639x+4.7535	0.9262	1.969	2.125±0.753
	福州	y=0.5661x + 4.8864	0.9664	1.222
	南平	y=0.4829x + 4.4215	0.9717	3.313
	宁德	y=1.0471x + 4.2766	0.986	1.995
98% 吡菌胺 98% Fluopicolide	龙岩	y=2.8279x + 7.1018	0.9467	0.476	0.370±0.064
	福州	y=1.7423x + 7.0719	0.9618	0.304
	南平	y=1.8045x + 6.9453	0.9748	0.340
	宁德	y=2.0973x + 7.1405	0.9205	0.360
95% 嘧菌酯 95% Azoxystrobin	龙岩	y=0.3594x+3.9249	0.9060	19.917	23.447±3.666
	福州	y=0.2455x+4.2175	0.9675	24.220
	南平	y=0.4168x+3.7405	0.9712	20.530
	宁德	y=0.4742x+3.4013	0.9047	29.120
94% 氰霜唑 94% Cyazofamid	龙岩	y=0.1778x+5.0507	0.9722	0.753	0.713±0.088
	福州	y=0.2725x+5.0668	0.9314	0.783
	南平	y=0.2603x + 5.0734	0.9046	0.754
	宁德	y=0.5180x+5.2986	0.9486	0.562
95% 烯酰吗啉 95% Dimethomorth	龙岩	y=3.5896x + 10.272	0.9738	0.230	0.239±0.011
	福州	y=3.3262x + 9.9644	0.9889	0.225
	南平	y=3.6195x + 10.017	0.9807	0.250
	宁德	y=3.6711x + 10.101	0.9776	0.249

下载: 导出CSV

参考文献(27)

[1]	叶泉清, 钟佳玲, 陈媚, 等. 槟榔芋疫霉菌生物学特性、致病力测定及田间防治药剂筛选 [J]. 南方农业学报, 2016, 47(4):588−593. doi: 10.3969/j:issn.2095-1191.2016.04.588 YE Q Q, ZHONG J L, CHEN M, et al. Biological characteristics, virulence of Phytophthora colocasiae Racib. from Colocasia esculenta L. var. cormosus Chang and fungicides screening for field control [J]. Journal of Southern Agriculture, 2016, 47(4): 588−593. (in Chinese) doi: 10.3969/j:issn.2095-1191.2016.04.588
[2]	兰成忠, 卢学松, 姚锦爱, 等. 芋疫霉菌的巢式PCR检测 [J]. 福建农业学报, 2019, 34(1):76−82. LAN C Z, LU X S, YAO J A, et al. Nested-PCR detection of taro leaf blight pathogen Phytophthora colocasiae [J]. Fujian Journal of Agricultural Sciences, 2019, 34(1): 76−82. (in Chinese)
[3]	NATH V S, HEGDE V M, JEEVA M L, et al. Morphological, pathological and molecular characterization of Phytophthora colocasiae responsible for taro leaf blight disease in India [J]. Phytoparasitica, 2015, 43(1): 21−35. doi: 10.1007/s12600-014-0422-5
[4]	孙道旺, 曹继芬, 裴卫华, 等. 云南魔芋新病害—疫病病原菌的鉴定 [J]. 植物病理学报, 2015, 45(1):84−87. SUN D W, CAO J F, PEI W H, et al. Identification of a new Phytophthora blight disease on konjac in Yunnan [J]. Acta Phytopathologica Sinica, 2015, 45(1): 84−87. (in Chinese)
[5]	王瑞仙, 邵梅, 于德才, 等. 魔芋疫病烟草疫霉菌(Phytophthora nicotianae)的致病性研究 [J]. 云南农业大学学报(自然科学), 2018, 33(2):213−217. WANG R X, SHAO M, YU D C, et al. Pathogenicity identification of Phytophthora nicotianae causing konjac disease in Yunnan, China [J]. Journal of Yunnan Agricultural University (Natural Science), 2018, 33(2): 213−217. (in Chinese)
[6]	董伟清, 何芳练, 江文, 等. 荔浦芋疫病病原鉴定及原生质体制备 [J]. 南方农业学报, 2016, 47(11):1861−1866. doi: 10.3969/jissn.2095-1191.2016.11.1861 DONG W Q, HE F L, JIANG W, et al. Identification of Colocasia esculenta(L. ) Schott var. Lipu Taro leaf blight disease causal pathogen and its protoplasts preparation [J]. Journal of Southern Agriculture, 2016, 47(11): 1861−1866. (in Chinese) doi: 10.3969/jissn.2095-1191.2016.11.1861
[7]	HE X L, MIYASAKA S C, FITCH M M M, et al. Taro (Colocasia esculenta) transformed with a wheat oxalate oxidase gene for improved resistance to taro pathogen Phytophthora colocasiae [J]. HortScience, 2013, 48(1): 22−27. doi: 10.21273/HORTSCI.48.1.22
[8]	MISRA R S, MISHRA A K, SHARMA K, et al. Characterisation ofPhytophthora colocasiaeisolates associated with leaf blight of taro in India [J]. Archives of Phytopathology And Plant Protection, 2011, 44(6): 581−591. doi: 10.1080/03235400903266339
[9]	KO W H. Mating-type distribution of Phytophthora colocasiae on the island of Hawaii [J]. Mycologia, 1979, 71(2): 434. doi: 10.1080/00275514.1979.12021021
[10]	LIN M J, KO W H. Occurrence of isolates of Phytophthora colocasiae in Taiwan with homothallic behavior and its significance [J]. Mycologia, 2008, 100(5): 727−734. doi: 10.3852/08-070
[11]	ZHANG K M, ZHENG F C, LI Y D, et al. Isolates of Phytophthora colocasiae from Hainan Island in China: Evidence suggesting an Asian origin of this species [J]. Mycologia, 1994, 86(1): 108. doi: 10.1080/00275514.1994.12026379
[12]	ANN P J, KAO C W, KO W H. Mating-type distribution of Phytophthora colocasiae in Taiwan [J]. Mycopathologia, 1986, 93(3): 193−194. doi: 10.1007/BF00443524
[13]	陆叶, 陆志翔, 何芳练, 等. 桂东南地区芋疫霉的交配型研究 [J]. 基因组学与应用生物学, 2013, 32(3):285−290. LU Y, LU Z X, HE F L, et al. A research on mating types of Phytophthora colocasiae in south-eastern Guangxi Province [J]. Genomics and Applied Biology, 2013, 32(3): 285−290. (in Chinese)
[14]	董伟清, 江文, 何芳练, 等. 广西地区荔浦芋新品种疫霉交配型和甲霜灵抗性研究 [J]. 长江蔬菜, 2018, (10):41−44. DONG W Q, JIANG W, HE F L, et al. Study on mating type and metalaxyl resistance of a new variety of taro in Lipu, Guangxi [J]. Journal of Changjiang Vegetables, 2018(10): 41−44. (in Chinese)
[15]	ADOMAKO J, KWOSEH C, MOSES E. Metalaxyl sensitivity and aggressiveness of phytophthora colocasiae isolates associated with taro leaf blight disease [J]. JOURNAL OF PLANT PATHOLOGY, 2017, 99(1): 205−210.
[16]	游世奇. 5种药剂对槟榔芋疫病的田间防效 [J]. 中国植保导刊, 2020, 40(2):71−72. doi: 10.3969/j.issn.1672-6820.2020.02.017 YOU S Q. Field control effect of five pesticides on betel nut taro blight [J]. China Plant Protection, 2020, 40(2): 71−72. (in Chinese) doi: 10.3969/j.issn.1672-6820.2020.02.017
[17]	方辉, 张惠琴, 陈孝赏, 等. 双炔酰菌胺防治红香芋疫病的效果及应用技术 [J]. 浙江农业科学, 2016, 57(6):899−900,911. FANG H, ZHANG H Q, CHEN X S, et al. Effect and application technology of diacetylbenzamide on controlling red taro blight [J]. Journal of Zhejiang Agricultural Sciences, 2016, 57(6): 899−900,911. (in Chinese)
[18]	李本金, 陈庆河, 兰成忠, 等. 福建省大豆根腐病病原菌的鉴定 [J]. 福建农业学报, 2011, 26(5):798−803. doi: 10.3969/j.issn.1008-0384.2011.05.024 LI B J, CHEN Q H, LAN C Z, et al. A identification and pathogenicity test of the pathogens causing soybean root rot in Fujian [J]. Fujian Journal of Agricultural Sciences, 2011, 26(5): 798−803. (in Chinese) doi: 10.3969/j.issn.1008-0384.2011.05.024
[19]	李华义, 何运转, 张艳杰, 等. 掌叶半夏疫病病原菌的分离与鉴定 [J]. 河北农业大学学报, 2016, 39(4):68−72. LI H Y, HE Y Z, ZHANG Y J, et al. Isolation and identification of blight disease on Pinellia pedatisecta [J]. Journal of Hebei Agricultural University, 2016, 39(4): 68−72. (in Chinese)
[20]	郑小波. 疫霉菌及其研究技术[M]. 北京: 中国农业出版社, 1997.
[21]	MBONG G A, FOKUNANG C N, FONTEM L A, et al. An overview of Phytophthora colocasiae of cocoyams: a potential economic disease of food security in Cameroon [J]. Discourse Journal of Agriculture & Food Sciences, 2013, 1(9): 140−145.
[22]	苗建强, 蔡萌, 张灿, 等. 植物病原卵菌对重要抑制剂的抗性分子机制研究进展 [J]. 农药学学报, 2019, 21(5-6):736−746. MIAO J Q, CAI M, ZHANG C, et al. Molecular resistance mechanism of phytopathogenic oomycete to several important fungicides [J]. Chinese Journal of Pesticide Science, 2019, 21(5-6): 736−746. (in Chinese)
[23]	路粉, 赵建江, 刘晓芸, 等. 马铃薯晚疫病菌对甲霜灵的抗性监测及替代药剂防治效果 [J]. 中国农业科学, 2018, 51(14):2700−2710. doi: 10.3864/j.issn.0578-1752.2018.14.007 LU F, ZHAO J J, LIU X Y, et al. Monitoring of resistance of Phytophthora infestans on potato to metalaxyl and the control efficacy of alternative fungicides [J]. Scientia Agricultura Sinica, 2018, 51(14): 2700−2710. (in Chinese) doi: 10.3864/j.issn.0578-1752.2018.14.007
[24]	蓝雯婷, 任董董, 李瑞环, 等. 荔枝霜疫霉对烯酰吗啉的抗性风险评估 [J]. 果树学报, 2023, 40(6):1226−1234. LAN W T, REN D D, LI R H, et al. Baseline sensitivity and resistance risk assessment of Peronophythora li-tchii to dimethomorph [J]. Journal of Fruit Science, 2023, 40(6): 1226−1234. (in Chinese)
[25]	张世才, 李怡斐, 王春萍, 等. 重庆地区辣椒疫霉交配型分布及对烯酰吗啉的敏感性现状 [J]. 植物保护, 2022, 48(2):173−176,182. ZHANG S C, LI Y F, WANG C P, et al. Distribution of mating types of Phytophthora capsici and its sensitivity to dimethomorph in Chongqing [J]. Plant Protection, 2022, 48(2): 173−176,182. (in Chinese)
[26]	邹芬, 何烈干, 李湘民, 等. 江西槟榔芋疫病病原菌鉴定及室内药剂筛选 [J]. 植物保护, 2023, 49(1):311−316. ZOU F, HE L G, LI X M, et al. Identification of the pathogen of Binglang taro leaf blight in Jiangxi Province and laboratory screening of fungicides [J]. Plant Protection, 2023, 49(1): 311−316. (in Chinese)
[27]	洪爱梅, 张海艳, 段云辉, 等. 9种杀菌剂防控芋疫病效果评价 [J]. 世界农药, 2022, 44(9):41−45. HONG A M, ZHANG H Y, DUAN Y H, et al. Evaluation on the control efficacy of nine fungicides against taro leaf blight [J]. World Pesticide, 2022, 44(9): 41−45. (in Chinese)