基于EPG技术分析茶树品种对茶小绿叶蝉的抗性及其相关的抗性物质
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浙江省自然科学基金项目(LY17C140002);浙江省大学生科技创新项目(2016R40915,2017R409055)


Resistance of tea cultivars to the tea green leafhopper analyzed by EPG technique and their resistance-related substances
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    摘要:

    为深入探讨假眼小绿叶蝉在茶树上的刺探行为,进而筛选合适的刺探电位(electrical penetration graph,EPG)参数作为评判茶树抗感性指标,遂比较紫鹃、宁波黄金芽和黄观音等11个国家级和特色茶树良种对该叶蝉抗感性,并解析茶树抗感性与主要化学成分的相关性。使用刺探电位仪检测了该叶蝉在这11个茶树品种上的多种刺探行为及其参数、以聚类分析等方法解析各品种上EPG参数而判断每品种对于叶蝉的抗感性;再以HPLC检测茶叶主要成分,分析抗感性与化学成分的关联程度。结果如下:(1)进一步明确了代表该叶蝉在茶树上取食行为的7种EPG波形及其可能的生物学意义,即Np、A、S、C、E、F和R波,解析了各种波形的特征和出现规则,认为E波、F波以及未确定生物学意义的R波是抗性相关波。(2)在5h测试期间,叶蝉在黄观音和龙井43品种上的刺探次数最多,紫鹃、宁波黄金芽和金观音上的次之,铁观音和安吉白茶上的最少;E波平均持续时间在铁观音和黄牡丹上最长,乌牛早和安吉白茶上次之,紫鹃上最短;F波平均持续时间在黄观音上最长,紫鹃、龙井43和宁波黄金芽上次之,金观音上最短。(3)以刺探次数、韧皮部取食波E波历期、韧皮部取食困难波F波历期为指标,将11个品种聚为2类,即第Ⅰ类:安吉白茶、乌牛早、紫牡丹、中茶108、黄牡丹和铁观音;第Ⅱ类为黄观音、龙井43、紫鹃、宁波黄金芽和金观音;抗性强弱为:第Ⅱ类 > 第Ⅰ类。(4)从感性至抗性品种,儿茶素类含量明显升高,且儿茶素总量与E波历期显著负相关(P < 0.05);咖啡碱、氨基酸和可溶性糖含量皆未与E波、F波历期明显相关。认为:E波和F波历期以及刺探次数可作为茶树抗感性的主要参数;R波可能与韧皮部取食抗性相关;儿茶素类是抗性相关物质。研究可为深入探讨茶树抗虫机理提供参考。

    Abstract:

    To thoroughly investigate probing behavior of the tea green leafhopper, Empoasca vitis (G the), on tea plants, determine adequate electrical penetration graph (EPG) parameters as reliable indices for measuring tea plant resistance/susceptibility levels to the leafhopper, and further analyze their potential correlations to the major defensive components in tea leaves, 11 national or regional elite/specificity cultivars, such as Zijuan, Ningbohuangjinya and Huangguangyin, were studied using EPG techniques. Various leafhopper probing behaviors and EPG parameters were analyzed to evaluate the resistance/susceptibility of every cultivar to the leafhopper. Major (primary) tea leaf chemical components were determined using high-pressure liquid chromatography. Our results showed that:1) seven distinct EPG waveforms (Np, A, S, C, E, F, and R) with possible biological significance during leafhopper feeding were recorded, and each waveform and its regular emergence sequence were analyzed. The waveforms E and F, plus the waveform R (with unknown biological significance) were considered to be the resistance-related waveforms. 2) During the 5-h testing period, the numbers of probing instances by the leafhopper on cultivars Huangguanyin and Longjing 43 were the highest, followed by those on cultivars Zijuan, Ningbohuangjinya and Jinguanyin, and the numbers on cultivars Tieguanyin and Anjibaicha were the lowest. The average durations of waveform E on cultivars Tieguanyin and Huangmudan were the longest, followed by the durations on cultivars Wuniuzao and Anjibaicha, and that on cultivar Zijuan was the shortest. The average duration of waveform F on cultivar Huangguanyin was the longest, followed by the durations on cultivars Zijuan, Longjing 43, and Ningbohuangjinya, and that on cultivar Jinguanyin was the shortest. 3) These 11 tested tea cultivars were classified into two groups via cluster analysis based on the number of probing instances, and durations of waveforms E and F as indices; group I:cultivars Anjibaicha, Wuniuzao, Zimudan, Zhongcha 108, Huangmudan, and Tieguanyin and group Ⅱ:cultivars Zijuan, Longjing 43, Ningbohuangjinya, Huangguanyin, and Jinguanyin. The resistance level of tea cultivars in group Ⅱ was stronger than that in group I. 4) From susceptible to resistant cultivars, the total catechin content rose, and it was negatively correlated with the duration of waveform E (P < 0.05); however, the contents of caffeine, amino acids and soluble sugars were not significantly correlated with the duration of waveform E or F. It is suggested that the number of probing instances, and durations of waveforms E and F are relatively reliable EPG indices for measuring tea plant resistance/susceptibility levels to the leafhopper. Waveform R might be correlated with feeding-resistance in phloem, but needs further investigation. Catechins seemed to be the key defensive compounds in tea leaves against the leafhopper feeding. This study provides in-depth reference information for further investigations on the resistance mechanisms of tea plants against pests.

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郑雨婷,王梦馨,崔林,韩善捷,俞鹏飞,韩宝瑜.基于EPG技术分析茶树品种对茶小绿叶蝉的抗性及其相关的抗性物质.生态学报,2017,37(23):8015~8028

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