Identification of the Pathogen of Chinese Chestnut Yellow Crinkle and Its Control

doi:10.13304/j.nykjdb.2021.0603

Abstract

Abstract:

To identify the pathogen of Chinese chestnut yellow crinkle （CnYC） and explore an effective comprehensive prevention and control technology， the pathogen was identified by the combination of morphology and molecular biology and agricultural antibiotics were screened by hanging bag infusion perforated in the field. The results showed that the pathogen causing Chinese chestnut yellow crinkle was Candidatus Phytoplasma castaneae belonged to 16SrXIX-A subgroup， and the strain number was CnYC-Hebei； the tetracycline hydrochloride showed the best control effect on CnYC， and its control rate was 85.3%. The control technology included as follows： tetracycline hydrochloride was infused into the diseased plant of CnYC in pre-deciduous stage， the diseased branches were pruned after defoliation stage， brassinolide and foliar fertilizer were sprayed on the leaves at the leaf development stage， and stinging insects were trapped and killed in growth stage. The control rate of comprehensive prevention and control technology reached 86.4%， the single fruit weight was 8.1 g， the full fruit rate was 91.9%， the chestnut size was 49.3 mm×42.3 mm， and the leaf size was 18.1 mm×7.5 cm. The indicators were all significantly better than the control group （P<0.05）. This technology not only effectively controled the disease， but also restored the production of diseased trees， which provided an effective comprehensive prevention and control technology for CnYC.

Key words: Chinese chestnut yellow crinkle, phytoplasma, identification, comprehensive control

摘要：

为明确引起板栗黄化皱缩病（Chinese chestnut yellow crinkle，CnYC）的病原菌及分类地位，探索一套有效的综合防控技术，利用形态学和分子生物学相结合的方法对引起该病害的病原菌进行了鉴定，并采用田间打孔吊袋输液的方法筛选农用抗生素。结果表明：引起板栗黄化皱缩病病原菌为Candidatus Phytoplasma castaneae，属于16SrXIX-A亚组，菌株编号为CnYC-Hebei；盐酸四环素对该病害防治效果较好，其防效达到85.3%。采收后落叶前利用盐酸四环素打孔吊袋输液方法，并结合落叶后修剪发病枝条、展叶期叶面喷施芸苔素内酯和叶面肥以及生长期诱杀刺吸式昆虫，从病害诊断、防病虫以及壮树3个方面集成了板栗黄化皱缩病三位一体的综合防控技术，对该病害进行了田间综合防控示范试验。调查分析显示，综合防控组的防治效果达到86.4%、单果重为8.1 g、饱果率为91.9%、栗蓬大小为49.3 mm×42.3 mm，叶片大小为18.1 cm×7.5 cm，这些指标均显著好于对照组（P<0.05），使发病植株恢复了生产，为开发安全有效的CnYC综合防控技术提供了参考。

关键词: 板栗黄化皱缩病, 植原体, 鉴定, 综合防控

CLC Number:

S436.6

Wenjie YANG, Xiaolei WEN, Lina FENG, Nana ZHANG, Weiming SUN, Huixia QI. Identification of the Pathogen of Chinese Chestnut Yellow Crinkle and Its Control[J]. Journal of Agricultural Science and Technology, 2022, 24(3): 148-156.

杨文杰, 温晓蕾, 冯丽娜, 张娜娜, 孙伟明, 齐慧霞. 板栗黄化皱缩病的病原菌鉴定及其防治[J]. 中国农业科技导报, 2022, 24(3): 148-156.

Figures/Tables 7

References 18

1	阚黎娜,李倩,谢爽爽,等.我国板栗种质资源分布及营养成分比较[J].食品工业科技, 2016, 37(20): 396-400.
	KAN L N, LI Q, XIE S S, et al.. Resource distribution and nutritional quality difference of Chinese chestnuts [J]. Sci. Technol. Food Industry, 2016, 37(20): 396-400.
2	郜付敏.基于神经网络模型的迁西板栗市场价格分析与预测[D].河北保定: 河北大学, 2020.
	GAO F M. Analysis and Forecast of Qianxi Chestnut Market Price Based on Neural Network Model [D]. Hebei Baoding: Hebei University, 2020.
3	LIN C L, LI H F, ZHANG G Z, et al. Molecular identification and characterization of a new phytoplasma strain associated with Chinese chestnut yellow crinkle disease in China [J]. Forest Pathology, 2011, 41(3):233-236.
4	张树航,王广鹏,郭燕,等. 板栗小叶病的症状及防治方法[J].河北果树, 2020, 3(2): 45.
	ZHANG S H, WANG G P, GUO Y, et al.. Symptoms and control methods of chestnut lobular disease [J]. Hebei Fruits, 2020, 3(2): 45.
5	LEE I M, HAMMOND R W, DAVIS R E, et a1. Universal amplification and analysis of pathogen 16S rDNA for classification and identification of mycoplasmalike organisms [J]. Phytopathology, 1993, 83(8): 834-842.
6	韩剑,陈杭,艾克热木·买买提,等.杏褪绿卷叶植原体新疆分离物核糖体蛋白基因的克隆与序列分析[J].生物安全学报,2021,30(1):36-42.
	HAN J, CHEN H, AIKEREMU M,et al.. Cloning and sequencing of the ribosomal protein gene of the phytoplasma of apricot chlorotic leaf roll in Xinjiang [J]. J. Biosafety, 2021,30(1):36-42.
7	廖晓兰,罗宽,朱水芳.植原体的分类及分子生物学研究进展[J]. 植物检疫,2002, 3(12): 41-46.
	LIAO X L, LUO K, ZHU S F. Phytoplasma classification and molecular biology research progress [J]. Plant Quarantine, 2002, 3(12): 41-46.
8	赖帆,李永,徐启聪,等. 植原体的最新分类研究动态[J]. 微生物学通报, 2008, 16(2): 291-295.
	LAI F, LI Y, XU Q C, et al.. The latest research trends of phytoplasma classification[J]. Bull. Microbiol., 2008, 16(2): 291-295.
9	JUNG H Y, TOSHIMI S, SHIGEYUKI K, et al. ‘Candidatus Phytoplasma castaneae’, a novel phytoplasma taxon associated with chestnut witches’ broom disease [J]. Int. J. Syst. Evol. Microbiol., 2002, 52(5):1543-1549.
10	JUNG H Y. ‘Candidatus Phytoplasma ziziphi’, a novel phytoplasma taxon associated with jujube witches’ broom disease [J]. Int. J. Syst. Evol. Microbiol., 2003, 53(4): 1037-1041.
11	林彩丽,朱晓清,李志朋,等.北京怀柔区板栗黄化皱缩病害调查[J].中国森林病虫, 2011, 30(5): 24-26.
	LIN C L, ZHU X Q, LI Z P, et al.. Investigation on yellowing and shrinking disease of Chinese chestnut in Huairou District, Beijing [J]. China Forest Dise. Insects, 2011, 30(5): 24-26.
12	朱自平,王殿宏,曹立娜.枣疯病发生规律及防治措施[J]. 现代农村科技, 2020, 3(8): 43-45.
	ZHU Z P, WANG D H, CAO L N. Occurrence law and control measures of jujube madness [J]. Modern Rural Sci. Technol., 2020, 3(8): 43-45.
13	李志朋.怀柔区板栗黄化皱缩病发生相关因子调查及综合防控示范[J].中国植保导刊. 2020,40(10):65-70.
14	郭建民,杨俊强,薛新平.枣疯病研究进展[J].山西农业科学, 2017, 45(8): 1389-1392.
	GUO J M, YANG J Q, XUE X P. Research progress of jujube madness [J]. Shanxi Agric. Sci., 2017, 45(8): 1389-1392.
15	欧阳敦君.永州市柑橘示范场黄龙病防治技术经验[J]. 湖南农业, 2020, 2(7): 16-19.
	OUYANG D J. Experience in prevention and control technology of Huanglongbing in citrus demonstration farm in Yongzhou city [J]. Hunan Agric., 2020, 2(7): 16-19.
16	李岩峰,邹大军.四环素与阿维菌素复配防治柳树植原体病害初探[J].林业科技通讯, 2015, (7): 47-48.
	LI Y F, ZOU D J. Preliminary study on combination of tetracycline and abamectin to control willow phytoplasma diseases [J]. Forestry Sci. Technol. Commun., 2015, (7): 47-48.
17	胡慧,蔡战伟.浅谈泡桐丛枝病的发生和防治技术[J].花卉, 2018, 2(18): 301-302.
18	赵进红,王玉山,刘静,等.枣疯病大田防治药物筛选研究[J]. 果树学报, 2010, 27(6): 980-986.
	ZHAO J H, WANG Y S, LIU J, et al.. Research on the screening of drugs for the prevention and control of jujube madness in the field [J]. J. Fruit Trees, 2010, 27(6): 980-986.

编号 Number	株系 Strain	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18
1	CnYC-Hebei	1.00
2	CP000061	0.52	1.00
3	AB054986	1.00	0.52	1.00
4	AF498307	0.74	0.54	0.48	1.00
5	AY197655	0.59	0.52	0.48	0.63	1.00
6	AY390261	0.69	0.57	0.52	0.69	0.89	1.00
7	AF092209	0.65	0.56	0.52	0.65	0.77	0.85	1.00
8	AF248956	0.71	0.60	0.58	0.67	0.80	0.89	0.74	1.00
9	AF248957	0.56	0.50	0.52	0.54	0.56	0.62	0.58	0.66	1.00
10	AJ542541	0.54	0.56	0.36	0.43	0.50	0.49	0.44	0.51	0.42	1.00
11	AB052873	0.68	0.63	0.61	0.68	0.68	0.76	0.78	0.76	0.61	0.66	1.00
12	AF248959	0.46	0.85	0.47	0.51	0.46	0.43	0.45	0.52	0.39	0.48	0.57	1.00
13	AF248960	0.41	0.65	0.51	0.43	0.45	0.40	0.41	0.48	0.35	0.55	0.52	0.70	1.00
14	AJ550984	0.66	0.59	0.51	0.66	0.73	0.81	0.74	0.74	0.61	0.66	0.87	0.51	0.49	1.00
15	AF147708	0.51	0.48	0.80	0.53	0.59	0.60	0.53	0.67	0.63	0.44	0.62	0.51	0.51	0.55	1.00
16	AY725228	0.24	0.54	0.25	0.28	0.26	0.25	0.29	0.27	0.21	0.28	0.31	0.62	0.44	0.27	0.27	1.00
17	AY725234	0.20	0.52	0.25	0.22	0.20	0.21	0.27	0.25	0.21	0.20	0.27	0.62	0.36	0.20	0.27	0.57	1.00
18	DQ174122	0.52	0.79	0.48	0.56	0.49	0.57	0.51	0.61	0.48	0.56	0.58	0.85	0.66	0.56	0.56	0.52	0.52	1.00

编号 Number	株系 Strain	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18
1	CnYC-Hebei	1.00
2	CP000061	0.52	1.00
3	AB054986	1.00	0.52	1.00
4	AF498307	0.74	0.54	0.48	1.00
5	AY197655	0.59	0.52	0.48	0.63	1.00
6	AY390261	0.69	0.57	0.52	0.69	0.89	1.00
7	AF092209	0.65	0.56	0.52	0.65	0.77	0.85	1.00
8	AF248956	0.71	0.60	0.58	0.67	0.80	0.89	0.74	1.00
9	AF248957	0.56	0.50	0.52	0.54	0.56	0.62	0.58	0.66	1.00
10	AJ542541	0.54	0.56	0.36	0.43	0.50	0.49	0.44	0.51	0.42	1.00
11	AB052873	0.68	0.63	0.61	0.68	0.68	0.76	0.78	0.76	0.61	0.66	1.00
12	AF248959	0.46	0.85	0.47	0.51	0.46	0.43	0.45	0.52	0.39	0.48	0.57	1.00
13	AF248960	0.41	0.65	0.51	0.43	0.45	0.40	0.41	0.48	0.35	0.55	0.52	0.70	1.00
14	AJ550984	0.66	0.59	0.51	0.66	0.73	0.81	0.74	0.74	0.61	0.66	0.87	0.51	0.49	1.00
15	AF147708	0.51	0.48	0.80	0.53	0.59	0.60	0.53	0.67	0.63	0.44	0.62	0.51	0.51	0.55	1.00
16	AY725228	0.24	0.54	0.25	0.28	0.26	0.25	0.29	0.27	0.21	0.28	0.31	0.62	0.44	0.27	0.27	1.00
17	AY725234	0.20	0.52	0.25	0.22	0.20	0.21	0.27	0.25	0.21	0.20	0.27	0.62	0.36	0.20	0.27	0.57	1.00
18	DQ174122	0.52	0.79	0.48	0.56	0.49	0.57	0.51	0.61	0.48	0.56	0.58	0.85	0.66	0.56	0.56	0.52	0.52	1.00

样点 Sample site	萌芽期 Germination stage		展叶期 Leafing stage		花期Flowering stage		成熟期Maturation stage		落叶前期Pre-deciduous stage
样点 Sample site	叶片Leaf	枝条 Branch	叶片Leaf	枝条 Branch	叶片Leaf	枝条 Branch	叶片Leaf	枝条 Branch	叶片Leaf	枝条 Branch
迁西杨家峪 Qianxi Yangjiayu	0	0	0	—	12.5	—	50.0	30.0	75.0	50.0
迁西尖山峪 Qianxi Jianshanyu	0	0	0	—	33.3	—	55.6	—	66.7	—
迁西上洪寨 Qianxi Shanghongzhai	0	0	0	—	14.2	—	42.9	36.4	57.1	54.6
平均检出率 Average rate	0	0	0	—	20.0	—	47.5	33.2	66.3	52.3

样点 Sample site	萌芽期 Germination stage		展叶期 Leafing stage		花期Flowering stage		成熟期Maturation stage		落叶前期Pre-deciduous stage
样点 Sample site	叶片Leaf	枝条 Branch	叶片Leaf	枝条 Branch	叶片Leaf	枝条 Branch	叶片Leaf	枝条 Branch	叶片Leaf	枝条 Branch
迁西杨家峪 Qianxi Yangjiayu	0	0	0	—	12.5	—	50.0	30.0	75.0	50.0
迁西尖山峪 Qianxi Jianshanyu	0	0	0	—	33.3	—	55.6	—	66.7	—
迁西上洪寨 Qianxi Shanghongzhai	0	0	0	—	14.2	—	42.9	36.4	57.1	54.6
平均检出率 Average rate	0	0	0	—	20.0	—	47.5	33.2	66.3	52.3

药剂名称 Chemical	病情指数 Disease index	防治效果Control effect/%	单果重 Weight per fruit/g	饱果率 Full fruit rate/%	栗蓬大小 Pod size/mm		叶片大小 Leaf size/cm
药剂名称 Chemical	病情指数 Disease index	防治效果Control effect/%	单果重 Weight per fruit/g	饱果率 Full fruit rate/%	纵径 Longitudinal diameter	横径 Horizontal diameter	叶长 Leaf length	叶宽 Leaf width
对照CK	88.7±7.5 a	0.0 d	4.4±0.1 d	30.3±3.1 e	34.42±0.9 c	28.2±2.8 c	14.4±2.2 b	5.3±1.3 c
氟苯尼考Florfenicol	71.0±9.9 a	19.9±19.3 c	6.3±0.1 b	48.3±4.5 d	37.6±3.0 bc	31.9±3.9 bc	14.5±0.7 b	5.9±0.3 bc
井冈霉素Validamycin	49.0±15.6 b	44.7±17.6 b	6.1±0.1 b	81.0±4.6 c	39.7±1.9 ab	33.7±5.7 ab	15.3±0.5 ab	6.2±0.1 b
氯霉素Chloramphenicol	46.3±6.5 b	47.8±7.3 b	5.8±0.1 e	86.3±7.1 bc	40.2±1.4 ab	35.3±6.7 ab	15.5±0.4 ab	5.8±0.2 bc
土霉素Oxytetracycin	42.0±13.5 b	52.6±15.2 b	5.8±0.1 e	85.7±3.5 bc	38.6±4.8 ab	32.5±4.5 bc	14.8±1.0 ab	5.4±1.0 c
罗红霉素Roxithromycin	33.3±11.6 bc	62.4±13.0 ab	7.1±0.1 ab	84.3±7.4 c	40.5±1.3 ab	35.3±6.9 ab	16.1±0.4 a	6.9±0.1 a
多肽四环素 Polypeptide tetracycline	33.3±13.5 bc	62.4±15.2 ab	7.2±0.1 ab	97.7±4.0 a	42.4±3.2 a	36.6±7.1 a	16.2±0.3 a	6.4±0.3 ab
盐酸四环素 Tetracycline hydrochloride	13.3±8.0 c	85.0±15.2 a	7.4±0.1 a	88.0±3.0 b	39.4±1.0 ab	35.1±1.0 ab	15.5±0.3 ab	6.8±0.7 ab