Isolation and Identification of Pathogen of Rubber Tree Phytophthora Leaf Fall Disease and Screening of Control Fungicides

doi:10.13304/j.nykjdb.2023.0101

Abstract

Abstract:

In the last 2 years， rubber tree abnormal leaf fall occurred over large area in Xishuangbanna rubber plantation， and the symptoms were similar to those of rubber tree Phytophthora leaf fall disease. In order to clarify the pathogenic species and screen fungicides for effective control of this disease， the pathogen was isolated and purified by tissue isolation and mycelia cusp purification method， and the pathogenicity was determined by Koch’s postulates test， morphological and physiological characteristics， rDNA-ITS and COX Ⅱ gene sequence analysis. The results showed that the pathogen caused rubber tree abnormal leaf fall was identified as Phytophthora botryosa， which was a new record pathogen of rubber tree Phytophthora leaf fall disease in China. The antibiotic activities of 15 fungicides was tested in vitro with mycelial growth rate method. 48% dimethomorph-cyazofamid SC （suspension concentrate）， 80% dimethomorph WG （water dispersible granule）， 30% metalaxyl-dimethomorph WG， 35% dimethomorph-cymoxanil SC， 30% floridomide-metalaxyl WG and 0.3% tetramycin AS （aqueoces solution） could effectively inhibit the mycelial growth of the pathogen， with EC₅₀ of 0.137 1， 0.175 9， 0.201 7， 0.257 7， 0.265 1， 0.351 0 mg·L^-1， respectively， which could be used as candidates for control of the disease in the field.

Key words: rubber tree, Phytophthora leaf fall, Phytophthora botryosa, pathogen identification, newly recorded species in China, fungicide screening

摘要：

近2年西双版纳植胶区橡胶树发生大面积落叶，病害症状与季风性落叶病相似。为弄清引起橡胶树落叶病害的致病菌并筛选出有效的防治药剂，采用组织分离法和菌丝尖端纯化法进行病原菌分离、纯化，结合柯赫氏法则、形态特征、生理学特性及病原菌rDNA-ITS、COX Ⅱ核苷酸序列分析对病原菌进行鉴定。结果表明，引起橡胶树落叶病害的致病菌为簇囊疫霉（Phytophthora botryosa），为我国橡胶树季风性落叶病致病菌新记录种。采用菌丝生长速率法测定15种杀菌剂对该病原菌的室内抑菌活性，结果表明，48%烯酰·氰霜唑悬浮剂、80%烯酰吗啉水分散粒剂、30%烯酰·甲霜灵水分散粒剂、35%烯酰·霜脲氰悬浮剂、30%氟吡菌胺·甲霜灵水分散粒剂、0.3%四霉素水剂均可有效抑制病原菌的菌丝生长，其EC₅₀分别为0.137 1、0.175 9、0.201 7、0.257 7、0.265 1和0.351 0 mg·L^-1，这些药剂均可用作该病害田间防治的备选药剂。

关键词: 橡胶树, 季风性落叶病, 簇囊疫霉, 病原菌鉴定, 中国新记录种, 防治药剂

CLC Number:

S796

Yuping SHI, Yixian LIU, Guowei LI, Yi TANG, Liming DAI, Lanlan LI, Zhiying CAI. Isolation and Identification of Pathogen of Rubber Tree Phytophthora Leaf Fall Disease and Screening of Control Fungicides[J]. Journal of Agricultural Science and Technology, 2023, 25(11): 114-122.

施玉萍, 刘一贤, 李国伟, 唐轶, 戴利铭, 李岚岚, 蔡志英. 橡胶树季风性落叶病病原菌的分离鉴定及其防治药剂筛选[J]. 中国农业科技导报, 2023, 25(11): 114-122.

Figures/Tables 8

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基因 Gene	引物名称 Primer name	引物序列 Primer sequence （5’-3’）	文献 Reference
ITS	ITS1	TCCGTAGGTGAACCTGCGG	［22‑23］
ITS	ITS4	TCCTCCGCTTATTGATATGC	［22‑23］
COX Ⅱ	FM82	TTGGCAATTAGGTTTTCAAGATCC	［24］
COX Ⅱ	FM78	ACAAATTTCACTACATTGTCC	［24］

基因 Gene	引物名称 Primer name	引物序列 Primer sequence （5’-3’）	文献 Reference
ITS	ITS1	TCCGTAGGTGAACCTGCGG	［22‑23］
ITS	ITS4	TCCTCCGCTTATTGATATGC	［22‑23］
COX Ⅱ	FM82	TTGGCAATTAGGTTTTCAAGATCC	［24］
COX Ⅱ	FM78	ACAAATTTCACTACATTGTCC	［24］

药剂 Fungicide	毒力回归方程 Virulence regression equation	EC₅₀/（mg·L^-1）	95%置信区间 95% confidence interval/（mg·L^-1）	相关系数 Correlation coefficient
48%烯酰·氰霜唑悬浮剂 48% Dimethomorph-cyazofamid SC	y=1.737 5x+1.499 4	0.137 1	0.075 5~0.339 5	0.906 4
80%烯酰吗啉水分散粒剂 80% Dimethomorph WG	y=1.156 7x+0.877 0	0.175 9	0.130 6~0.255 2	0.971 1
30%烯酰·甲霜灵水分散粒剂 30% Metalaxyl-dimethomorph WG	y=2.105 3x+1.463 6	0.201 7	0.173 0~0.234 9	0.999 4
35%烯酰·霜脲氰悬浮剂 35% Dimethomorph-cymoxanil SC	y=0.943 7x+0.555 6	0.257 7	0.170 5~0.485 3	0.948 5
30%氟吡菌胺·甲霜灵水分散粒剂 30% Floridomide-metalaxyl WG	y=1.944 2x+1.121 0	0.265 1	0.225 6~0.318 2	0.946 2
0.3%四霉素水剂 0.3% Tetramycin AS	y=0.686 7x+0.312 3	0.351 0	0.225 5~0.633 7	0.992 4
25%甲霜·霜霉威可湿性粉剂 25% Metalaxyl-propamocarb WP	y=1.037 8x+0.080 0	0.837 4	0.581 5~1.442 8	0.988 0
72%甲霜·锰锌可湿性粉剂 72% Metalaxyl mancozeb WP	y=1.097 1x-0.568 2	3.295 2	2.400 8~4.606 0	0.982 9
560 g·L^-1嘧菌·百菌清悬浮剂 560 g·L^-1 Azoxystrobin-chlorothalonil SC	y=1.527 0x-2.519 5	44.659 1	21.861 5~61.597 8	0.999 5
77%硫酸铜钙可湿性粉剂 77% Copper calcium sulphate WP	y=1.291 3x-2.479 3	83.159 4	37.833 5~222.932 6	0.902 3
450 g·L^-1咪鲜胺水乳剂 450 g·L^-11 Prochloraz EW	y=1.949 4x-3.891 6	99.147 2	80.757 7~117.929 8	0.942 4
98%噁霉灵可溶粉剂 98% Hymexazol SP	y=1.910 0x-4.563 5	245.086 4	202.618 6~294.082 2	0.942 9
80%三乙膦酸铝可湿性粉剂 80% Phosethyl-Al WP	y=1.263 9x-3.682 3	819.421 7	556.526 5~1 558.901 1	0.989 6
250 g·L^-1 嘧菌酯悬浮剂 250 g·L^-1 Azoxystrobin SC	y=1.487 0x-4.522 6	1 100.288 6	562.785 0~3 238.667 2	0.910 9
500 g·L^-1 多菌灵悬浮剂 500 g·L^-1 Carbendazim SC	y=0.694 4x-2.996 5	20 671.976 3	10 875.421 0~65 490.655 0	0.911 7

药剂 Fungicide	毒力回归方程 Virulence regression equation	EC₅₀/（mg·L^-1）	95%置信区间 95% confidence interval/（mg·L^-1）	相关系数 Correlation coefficient
48%烯酰·氰霜唑悬浮剂 48% Dimethomorph-cyazofamid SC	y=1.737 5x+1.499 4	0.137 1	0.075 5~0.339 5	0.906 4
80%烯酰吗啉水分散粒剂 80% Dimethomorph WG	y=1.156 7x+0.877 0	0.175 9	0.130 6~0.255 2	0.971 1
30%烯酰·甲霜灵水分散粒剂 30% Metalaxyl-dimethomorph WG	y=2.105 3x+1.463 6	0.201 7	0.173 0~0.234 9	0.999 4
35%烯酰·霜脲氰悬浮剂 35% Dimethomorph-cymoxanil SC	y=0.943 7x+0.555 6	0.257 7	0.170 5~0.485 3	0.948 5
30%氟吡菌胺·甲霜灵水分散粒剂 30% Floridomide-metalaxyl WG	y=1.944 2x+1.121 0	0.265 1	0.225 6~0.318 2	0.946 2
0.3%四霉素水剂 0.3% Tetramycin AS	y=0.686 7x+0.312 3	0.351 0	0.225 5~0.633 7	0.992 4
25%甲霜·霜霉威可湿性粉剂 25% Metalaxyl-propamocarb WP	y=1.037 8x+0.080 0	0.837 4	0.581 5~1.442 8	0.988 0
72%甲霜·锰锌可湿性粉剂 72% Metalaxyl mancozeb WP	y=1.097 1x-0.568 2	3.295 2	2.400 8~4.606 0	0.982 9
560 g·L^-1嘧菌·百菌清悬浮剂 560 g·L^-1 Azoxystrobin-chlorothalonil SC	y=1.527 0x-2.519 5	44.659 1	21.861 5~61.597 8	0.999 5
77%硫酸铜钙可湿性粉剂 77% Copper calcium sulphate WP	y=1.291 3x-2.479 3	83.159 4	37.833 5~222.932 6	0.902 3
450 g·L^-1咪鲜胺水乳剂 450 g·L^-11 Prochloraz EW	y=1.949 4x-3.891 6	99.147 2	80.757 7~117.929 8	0.942 4
98%噁霉灵可溶粉剂 98% Hymexazol SP	y=1.910 0x-4.563 5	245.086 4	202.618 6~294.082 2	0.942 9
80%三乙膦酸铝可湿性粉剂 80% Phosethyl-Al WP	y=1.263 9x-3.682 3	819.421 7	556.526 5~1 558.901 1	0.989 6
250 g·L^-1 嘧菌酯悬浮剂 250 g·L^-1 Azoxystrobin SC	y=1.487 0x-4.522 6	1 100.288 6	562.785 0~3 238.667 2	0.910 9
500 g·L^-1 多菌灵悬浮剂 500 g·L^-1 Carbendazim SC	y=0.694 4x-2.996 5	20 671.976 3	10 875.421 0~65 490.655 0	0.911 7

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