板栗黄化皱缩病检测方法优化及10个板栗品种抗性鉴定

doi:10.13304/j.nykjdb.2023.0310

中国农业科技导报 ›› 2024, Vol. 26 ›› Issue (6): 226-233.DOI: 10.13304/j.nykjdb.2023.0310

• 方法与技术创新 • 上一篇

板栗黄化皱缩病检测方法优化及10个板栗品种抗性鉴定

郭思柔¹^,²(), 温晓蕾¹^,²(), 栗佳宁¹^,², 王建华¹^,², 孙伟明¹^,², 冯丽娜¹^,², 齐慧霞¹^,²()

^1.河北科技师范学院，板栗产业技术教育部工程研究中心，河北秦皇岛 066004
^2.河北科技师范学院农学与生物科技学院，河北省作物逆境生物学重点实验室（筹），河北昌黎 066600

收稿日期:2023-04-19 接受日期:2023-10-07 出版日期:2024-06-15 发布日期:2024-06-12
通讯作者: 齐慧霞
作者简介:郭思柔 E-mail：1808456005@qq.com
温晓蕾 E-mail：xiaoleiwen@sina.com第一联系人：郭思柔和温晓蕾为共同第一作者。
基金资助:
国家重点研发计划项目(2020YED1000700);河北省教育厅平台项目(河北省板栗产业协同创新中心-2021)

Optimization of Chinese Chestnut Yellow Crinkle Detection Method and Resistance Identification of 10 Chestnut Varieties

Sirou GUO¹^,²(), Xiaolei WEN¹^,²(), Jianing LI¹^,², Jianhua WANG¹^,², Weiming SUN¹^,², Lina FENG¹^,², Huixia QI¹^,²()

^1.Engineering Research Center of Chestnut Industry Technology of Ministry of Education，Hebei Normal University of Science and Technology，Hebei Qinhuangdao 066004，China
^2.Key Laboratory of Crop Stress Biology of Hebei Province； College of Agronomy and Biotechnology，Hebei Normal University of Science and Technology，Hebei Changli 066600，China

Received:2023-04-19 Accepted:2023-10-07 Online:2024-06-15 Published:2024-06-12
Contact: Huixia QI

摘要/Abstract

摘要：

为优化板栗黄化皱缩病检测方法，明确10个板栗品种的抗性，通过对巢式PCR引物添加量、退火温度的优化，结合植原体检出率和田间病情指数进行抗性评价。结果表明，巢式PCR在引物添加量为1.0 μL、退火温度为54 ℃时，板栗植原体检出率最稳定；在毒源树嫁接400 d后，供试板栗品种均检测出植原体，其中‘燕山早丰芽变’叶片中植原体检出率最高，发病情况最严重，‘燕丽’叶片中植原体检出率最低，发病情况最轻；相关分析表明，植原体检出率和田间病情指数呈极显著正相关，相关系数为0.907，其线性回归方程为Y=0.276X+1.610；根据2种评价方法将供试板栗品种分为高抗、中抗和高感品种，其中‘燕秋’‘燕紫’‘燕龙’‘燕丽’为高抗品种，‘燕山早丰芽变’为高感品种。研究初步明确了10个板栗品种对黄化皱缩病的抗性差异，为选用抗性品种提供了基础数据。

关键词: 板栗, 植原体, 检测方法优化, 抗性鉴定

Abstract:

In order to optimize the detection method of Chinese chestnut yellow crinkle and clarify the resistance of 10 chestnut varieties， the resistance evaluation was carried out by optimizing the content of nested PCR primers and annealing temperature， combined with phytoplasma detection rate and field disease index. The results showed that the detection rate of chestnut phytoplasma was the most stable when the primer addition amount was 1.0 μL and the annealing temperature was 54 ℃. After 400 d of grafting， phytoplasmas were detected in all the tested chestnut varieties. Among them， the detection rate of phytoplasma in the leaves of ‘Yanshan Zaofeng Yabian’ was the highest and the incidence was the most serious， while the detection rate of phytoplasma in the leaves of ‘Yanli’ was the lowest and the incidence was the lightest. The correlation analysis showed that the detection rate of phytoplasma was significantly positively correlated with the field disease index， the correlation coefficient was 0.907， and the linear regression equation was Y=0.276X+1.610. According to 2 evaluation methods， the tested chestnut varieties were divided into high resistance， medium resistance and high sensitivity. Among them，‘Yanqiu’‘Yanzi’‘Yanlong’and‘Yanli’were high resistance varieties， and ‘Yanshan Zaofeng Yabian’was high sensitivity varieties. The resistance difference of 10 chestnut varieties to Chinese chestnut yellow crinkle was preliminarily clarified， which provided basic data for the selection of resistant varieties.

Key words: Castanea mollissima Blume, phytoplasma, detection method optimization, resistance identification

中图分类号:

S664.2

郭思柔, 温晓蕾, 栗佳宁, 王建华, 孙伟明, 冯丽娜, 齐慧霞. 板栗黄化皱缩病检测方法优化及10个板栗品种抗性鉴定[J]. 中国农业科技导报, 2024, 26(6): 226-233.

Sirou GUO, Xiaolei WEN, Jianing LI, Jianhua WANG, Weiming SUN, Lina FENG, Huixia QI. Optimization of Chinese Chestnut Yellow Crinkle Detection Method and Resistance Identification of 10 Chestnut Varieties[J]. Journal of Agricultural Science and Technology, 2024, 26(6): 226-233.

图/表 10

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基因 Gene	引物名称 Primer name	引物序列 Primer sequence（5’-3’）
16S rDNA	R16mF2	CATGCAAGTCGAACGGA
	R16mR1	CTTAACCCCAATCATCGAC
	R16F2n	GAAACGACTGCTAAGACTGG
	R16R2	TGACGGGCGGTGTGTACAAACCCCG

基因 Gene	引物名称 Primer name	引物序列 Primer sequence（5’-3’）
16S rDNA	R16mF2	CATGCAAGTCGAACGGA
	R16mR1	CTTAACCCCAATCATCGAC
	R16F2n	GAAACGACTGCTAAGACTGG
	R16R2	TGACGGGCGGTGTGTACAAACCCCG

板栗接穗品种 Chestnut scion variety	植原体检出率 DRP/%
板栗接穗品种 Chestnut scion variety	30 d	150 d	400 d
燕山早丰芽变 Yanshan Zaofeng Yabian	6.67 ABbc	63.33 Aa	73.33 Aa
早短优系 Zaoduan Youxi	13.33 ABab	50.00 ABab	56.67 ABab
燕青 Yanqing	10.00 ABabc	33.33 ABbc	53.33 ABCb
燕宁 Yanning	10.00 ABabc	36.67 ABbc	50.00 ABCbc
燕宝 Yanbao	3.33 Bbc	33.33 ABbc	50.00 ABCbc
燕康 Yankang	10.00 ABabc	36.67 ABbc	46.67 BCbcd
燕秋 Yanqiu	6.67 ABbc	40.00 ABbc	40.00 BCbcd
燕紫 Yanzi	0.00 Bc	30.00 ABbc	40.00 BCbcd
燕龙 Yanlong	13.33 ABab	30.00 ABbc	33.33 BCcd
燕丽 Yanli	20.00 Aa	23.33 Bc	30.00 Bd

板栗接穗品种 Chestnut scion variety	植原体检出率 DRP/%
板栗接穗品种 Chestnut scion variety	30 d	150 d	400 d
燕山早丰芽变 Yanshan Zaofeng Yabian	6.67 ABbc	63.33 Aa	73.33 Aa
早短优系 Zaoduan Youxi	13.33 ABab	50.00 ABab	56.67 ABab
燕青 Yanqing	10.00 ABabc	33.33 ABbc	53.33 ABCb
燕宁 Yanning	10.00 ABabc	36.67 ABbc	50.00 ABCbc
燕宝 Yanbao	3.33 Bbc	33.33 ABbc	50.00 ABCbc
燕康 Yankang	10.00 ABabc	36.67 ABbc	46.67 BCbcd
燕秋 Yanqiu	6.67 ABbc	40.00 ABbc	40.00 BCbcd
燕紫 Yanzi	0.00 Bc	30.00 ABbc	40.00 BCbcd
燕龙 Yanlong	13.33 ABab	30.00 ABbc	33.33 BCcd
燕丽 Yanli	20.00 Aa	23.33 Bc	30.00 Bd

板栗接穗品种 Chestnut scion variety	发病率 Incidence rate/%	田间病情指数 FDI
燕山早丰芽变 Yanshan Zaofeng Yabian	67.50±2.50 Aa	20.00±1.25 Aa
早短优系 Zaoduan Youxi	62.50±2.50 Aba	18.13±0.63 Aa
燕青 Yanqing	62.50±2.50 Aba	18.13±0.63 Aa
燕宁 Yanning	55.00±5.00 BCb	18.13±0.63 Aa
燕秋 Yanqiu	50.00±5.00 CDb	14.38±0.63 Bb
燕宝 Yanbao	52.50±2.50 BCDb	14.38±0.63 Bb
燕康 Yankang	52.50±2.50 BCDb	13.13±0.63 Bbc
燕紫 Yanzi	42.50±2.50 Dc	11.87±0.63 BCcd
燕龙 Yanlong	40.00±0.00 EFc	10.00±0.00 CDde
燕丽 Yanli	32.50±2.50 Fd	8.75±0.00 De

板栗黄化皱缩病检测方法优化及10个板栗品种抗性鉴定

Optimization of Chinese Chestnut Yellow Crinkle Detection Method and Resistance Identification of 10 Chestnut Varieties

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参考文献 22

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Metrics

板栗接穗品种 Chestnut scion variety	田间病情指数 FDI	植原体检出率 DRP/%
燕山早丰芽变 Yanshan Zaofeng Yabian	20.00	73.33
早短优系 Zaoduan Youxi	18.13	56.67
燕青 Yanqing	18.13	53.33
燕宁 Yanning	18.13	50.00
燕宝 Yanbao	14.38	50.00
燕康 Yankang	13.13	46.67
燕秋 Yanqiu	14.38	40.00
燕紫 Yanzi	11.87	40.00
燕龙 Yan Long	10.00	33.33
燕丽 Yanli	8.75	30.00
相关系数 Coefficient of association	0.907^**

抗性分级 Resistance grading	高抗HR	中抗MR	高感HS
植原体检出率 DRP/%	0<DRP≤40	40<DRP≤60	DRP≥60
田间病情指数 FDI	1.61<FDI≤12.65	12.65<FDI≤18.17	FDI>18.17

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