显著调控假眼小绿叶蝉行为的迷迭香挥发物鉴定

茶园生态系中生长着众多草本和木本植物,迷迭香就是其中常见的木本杂草。以其幼嫩部分为味源、茶园重要害虫假眼小绿叶蝉为试虫,用Y形管嗅觉仪做行为测定,以洁净空气为CK,发现随着味源剂量增加,其引诱的假眼小绿叶蝉数增至一个数值再减少,二者呈抛物线关系。从迷迭香挥发物中鉴定出46种化合物,包含醇类9种、酮类4种、酯类4种、酸类1种、烯烃类18种、烷烃类7种、醚类1种、醛类1种和酚类1种。醇类含量最大,占总量52.20%;单组分中桉树脑、左旋α-蒎烯和马鞭草烯酮的含量最大,分别占总量的42.54%、10.37%和6.82%。使用迷迭香挥发物中含量较大的25种组分为味源,每味源配制几种剂量,以正己烷为CK,用Y形管嗅觉仪进行行为测定,发现10-2g/mL樟脑、10-4g/mL石竹烯、10-6g/mL和10-8g/mLα-水芹烯、10-4g/mL和10-10g/mLα-松油醇、10-10g/mL和10-8g/mL桉树脑显著引诱假眼小绿叶蝉;而10-10g/mLβ-蒎烯显著排斥假眼小绿叶蝉。经分析认为:茶园中除了茶树之外,还有其它植物挥发物引诱假眼小绿叶蝉,可为筛选茶园中诱虫植物提供参考。     

假眼小绿叶蝉对茶梢挥发物的行为反应

【目的】研究假眼小绿叶蝉Empoasca vitis G?the对健康茶梢和蝉害茶梢挥发物的行为反应,筛选出可引诱假眼小绿叶蝉的茶梢挥发物。【方法】以动态吸附法收集健康茶梢和蝉害茶梢挥发物,用气相色谱–质谱联用仪(GC-MS)进行定性定量分析,借助于Y形嗅觉仪检测多种挥发物引诱假眼小绿叶蝉成虫的活性。【结果】从健康茶梢和蝉害茶梢中共鉴定出30种挥发物组分,其中烯烃类含量较大。健康茶梢和蝉害茶梢挥发物中共有组分有13种,蝉害之后其含量皆上升,其中Z-b-罗勒烯和乙酸叶醇酯的含量分别是健康茶梢中的142.27倍、12.90倍。蝉害茶梢中新出现的组分有12种,其中紫苏烯含量较高。在10-2、10~(-4)、10-6 g/mL浓度下,乙酸叶醇酯表现出极显著的引诱水平(P<0.01);紫苏烯在10~(-2)和10~(-4) g/mL浓度下表现出极显著引诱水平(P<0.01);10~(-4) g/mL浓度下,Z-b-罗勒烯和D-柠檬烯极显著引诱假眼小绿叶蝉(P<0.01);10~(-4) g/mL芳樟醇、10~(-6) g/mL乙酸正丁酯和10~(-6) g/mL D-柠檬烯呈现出显著引诱水平(P<0.05);混合物组分Blend1和Blend2分别表现出极显著和显著引诱水平;而10~(-6) g/mLa-法尼烯显著排斥假眼小绿叶蝉(P<0.05)。【结论】假眼小绿叶蝉成虫对健康茶梢和蝉害茶梢挥发物多种组分具有不同的行为反应,引诱效果较强的单组分或混合组分的选定可为田间引诱效果试验提供参考。     

Analysis of resistance in tomato and sweet pepper against the greenhouse whitefly using electrically monitored and visually observed probing and feeding behaviour

Different types of plant resistance against the greenhouse whitefly, Trialeurodes vaporariorum, were distinguished according to the tissue location of the resistance factors. The effects of resistance factors were compared by measuring the electrically monitored and visually observed probing and feeding behaviour of whiteflies on two resistant tomato genotypes (82207 and 82216), a susceptible tomato cultivar, and a non-host plant, sweet pepper. On sweet pepper, whiteflies displayed very short first probes, very long pathway probing and spent much time on non-feeding activities such as walking and standing still. Also, a high percentage of whiteflies rejected sweet pepper without ingesting substances from the phloem vessel. These data suggest a strong resistance that is based on the factors present in surface/epidermis and/or mesophyll layers of this plant. The behaviour of whiteflies on tomato 82207 was very different from that on sweet pepper. On tomato 82207 whiteflies apparently did not perceive resistance factors on the leaf surface and in the mesophyll. Resistance factors appeared to be present in the phloem tissue, because a higher number of phloem phases, longer phloem salivation periods and shorter phloem ingestion periods were observed when compared with the susceptible tomato cultivar. Tomato 82216 showed no clear resistance factor in the phloem. The importance of combined EPG and behavioural research for host-plant resistance studies are discussed.     

迷迭香挥发物不同组合对假眼小绿叶蝉行为的调控

为探讨茶园群落中非茶植物气味引诱或驱避重要茶树害虫假眼小绿叶蝉的效应,遂将茶园常见杂草迷迭香挥发物中樟脑、α-松油醇和石竹烯分别配成10-2g/m L剂量,再等量组成混合物,在每个橡皮头上载100#L制成诱芯,附于素馨黄粘板上,于茶园中诱捕假眼小绿叶蝉,发现这种三组分诱芯明显地驱避假眼小绿叶蝉。将行为反应中分别对假眼小绿叶蝉呈现显著引诱效应的10-2g/m L樟脑、10-4g/m L石竹烯、10-6g/m Lα-水芹烯、10-4g/m Lα-松油醇和10-8g/m L桉树脑等量配成五组分混合物,同法制成诱芯,附于素馨黄粘板,诱捕假眼小绿叶蝉。发现五组分诱芯:(1)显著引诱假眼小绿叶蝉,诱效稍强于现价段茶园中使用的源于茶梢挥发物的诱芯;(2)每日诱捕的假眼小绿叶蝉数与天数呈抛物线关系;(3)加入液体石蜡作为缓释剂,可将诱效的半衰期延长2.5—4.0d;(4)18:00—6:00诱得的假眼小绿叶蝉数量占全天的70%—75%。分析认为:非茶植物迷迭香的挥发物中含有显著引诱假眼小绿叶蝉信息物质,这类信息物质制成的五组分诱芯的诱效较强,可以作为一种防治假眼小绿叶蝉的手段,早晨和黄昏是最佳诱捕时分。     

茶树花挥发物对叶蝉三棒缨小蜂的引诱增强效应

茶树(Camellia sinensis)是我国重要的经济作物。小贯小绿叶蝉(Empoasca onukii)是为害茶树的主要害虫之一。叶蝉三棒缨小蜂(Stethynium empoascae)是小贯小绿叶蝉的主要卵寄生蜂,但茶树花对叶蝉三棒缨小蜂的引诱机制尚不明确。基于析因设计的方差分析结果显示,茶树花可引诱天敌叶蝉...     

茶园间作柑桔杨梅或吊瓜对叶蝉及蜘蛛类群数量和空间格局的影响

为评价茶园间作几种常见经济作物对重要害虫假眼小绿叶蝉及其主要天敌蜘蛛类群数量和空间格局的影响,遂选乌牛早品种纯茶园、乌牛早分别与柑桔、杨梅和吊瓜的间作茶园、以及安吉白茶与吊瓜间作茶园,2007年9月上旬—2008年12月下旬,每旬1次调查茶丛上、中、下层叶蝉和各种蜘蛛的数量。结果表明:(1)与纯茶园相比,间作茶园叶蝉种群数量和蜘蛛类群个体数量显著地增加,间作茶园蜘蛛种数显著地增加;(2)间作茶园茶丛上、中、下层叶蝉、蜘蛛个体数量分布明显区别于纯茶园茶丛上、中、下层叶蝉、蜘蛛个体数量分布;(3)茶丛上层的嫩梢是制作高档茶的原料,而纯茶园茶丛上层叶蝉虫口百分率为54.16%,间作茶园茶丛上层叶蝉虫口百分率皆减小,并且叶蝉高峰期间蜘蛛的跟随效应增强;(4)间作增加了经济收入并减少了防治次数。认为:(1)间作可在一定程度上调控叶蝉种群、蜘蛛类群的数量和空间格局;(2)间作可减轻叶蝉为害造成的产值损失,增强了茶园群落对于叶蝉的自然控制潜能。     

茶树CsGGDPS7基因的克隆和表达分析

从茶树基因组鉴定得到牻牛儿基牻牛儿基焦磷酸合成酶基因CsGGDPS7,以铁观音茶树为材料克隆获得全长cDNA(GenBank登录号MH891780)。CsGGDPS7序列全长1 185bp,包含1 098bp开放阅读框(ORF),编码365个氨基酸。预测结果显示在N端包含叶绿体转运肽,亚细胞定位于叶绿体中。氨基酸序列分析结果显示,具有类异戊二烯合成酶家族的5个保守域和2个特征功能域(DDXXXXD和DDXXD)。进化树结果表明与油橄榄(Olea europaea)OeGGDPS7亲缘关系最近。荧光定量检测显示,CsGGDPS7在叶片中的表达量显著高于其他组织,但花发育阶段CsGGDPS7表达量差异不显著。在乌龙茶做青过程中,CsGGDPS7在晒青阶段就快速上调表达并在三摇达到峰值。CsGGDPS1和CsGGDPS2表达也受做青调控,但CsGGDPS4不受做青调控。研究推测,CsGGDPS7可能与乌龙茶萜类香气物质合成密切相关。     

电子取食监测仪在植物抗虫性研究中的应用

主要综述了电子取食监测仪在植物对同翅目昆虫抗虫性研究中的应用情况.利用电子取食监测仪可监测同翅目昆虫（蚜虫、飞虱、叶蝉、蓟马、粉虱等）在不同抗、感虫品种上的取食行为,并根据取食行为参数如刺探频率、唾液分泌时间、韧皮部取食时间等在不同抗、感品种上的差异来区别品种的抗、感虫特性和确定抗虫因子存在的部位.     

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

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。     

Acceptability of different species of Brassicaceae as hosts for the cabbage aphid

The probing and feeding behaviour of the cabbage aphid, Brevicoryne brassicae (L.), (Homoptera, Aphididae) was studied on several plant species that represented various levels of acceptability: Sinapis alba L. (a permanent host plant), Capsella bursa-pastoris (L.) Med., Thlaspi arvense L., Lunaria annua L., Erysimum cheiranthoides L. (accidental host plants), Vicia faba L. (a non-host plant), using the electrical penetration graph technique (EPG). B. brassicae on V. faba did not show any patterns related to penetration of phloem vessels. Stylet penetration was deterred on L. annua and E. cheiranthoides where non-penetration prevailed, the periods of sap ingestion were short or did not occur, the percentage of time spent in the phloem was consistently low (5–6%) and E1 salivation predominated. The pathway activities were not suppressed on C. bursa-pastoris and T. arvense and the aphids spent an average of 3 h in the phloem during the 8-h experiment. However, a considerable delay between finding and accepting the phloem and a substantial proportion of E1 salivation (20–30% of all phloem activities) indicated a deterrent factor in the sieve elements of these plants. Aphid probing and sap ingestion were rarely interrupted on S. alba. The results of this study suggest that the deterrent agents vary in activity and may hinder stylet penetration at different levels (epidermis, parenchymatous tissues and/or phloem elements), depending on the plant species.     

秋末苏南茶园昆虫的群落组成及其趋色性

苏南地区名茶荟萃,而虫害历来较为严重。秋末选丹阳市一片无公害茶园,使用纯白、桃红、墨绿、果绿、湖蓝、天蓝、素馨黄、芽绿、土黄、桔黄、大红和紫色12种粘性色板诱虫。结果表明:① 4日内捕获7目42科85种30455头昆虫,其中优势类群是同翅目、膜翅目和双翅目,三者个体数分别占总个体数的86.5%、8.8%和2.5%。②主要害虫是假眼小绿叶蝉和茶蚜,分别占总个体数的15%和71.5%。③ 捕获的中华蜜蜂占总个体数8.2%。④捕获天敌昆虫1034头,其中,瓢虫类32头,占3.1%,主要种类是异色瓢虫和黄斑盘瓢虫;草蛉类582头,占56.3%,包括中华草蛉、大草蛉和丽草蛉;伞裙追寄蝇和蚕饰腹寄蝇等5种寄生蝇类111头,占10.7%;门氏食蚜蝇和黑带食蚜蝇等7种食蚜蝇类110头,占10.6%;螟蛉瘤姬蜂和花胸姬蜂等5种姬蜂类、茶尺蠖绒茧蜂和单白绵绒茧蜂等7种茧蜂类个体数分别占3.9%和9.4%。⑤ 芽绿、素馨黄和桔黄色板显著地引诱假眼小绿叶蝉和茶蚜;芽绿、素馨黄色板显著地引诱姬蜂和茧蜂;芽绿、土黄和果绿色板明显地引诱草蛉类;果绿、天蓝和紫色色板引诱较多的蝇类和蚊类等双翅目昆虫;素馨黄引诱的各类昆虫种数最多;纯白板上各类昆虫的多样性指数最大,表明其对许多昆虫都有引诱效应。秋末时节查明即将越冬的害虫和天敌昆虫种类、数量和益害比、以及优势种害虫和优势种天敌数量,探明多种色彩引诱益、害虫的差异,对于有效实施无公害封园防治,以压低越冬基数有重要意义。     

色彩对茶园昆虫的引诱力

为评估色板用于监测和诱捕茶园害虫的可能性,于10月份茶园昆虫盛发期在绍兴和杭州分别选有机、无公害茶园,同时使用10种粘性色板大面积诱捕茶园昆虫和蜘蛛.每种色彩重复10次.结果表明:(1)在两片供试茶园捕获的目、科和种类基本相同.蜘蛛很少.24 h后,绍兴茶园中10种色板共捕获9目53科10200个昆虫和蜘蛛;72 h 后杭州茶园中捕获10目63科21526个昆虫和蜘蛛.(2)同翅目为优势类群.绍兴茶园中,纯白色板诱捕的同翅目种数占该色板捕获的总物种数的15.2%、同翅目个体数占该色板捕获的总个体数的34.1%;其它9种色板诱捕同翅目种数占各自捕获总种数的21.2%～31.0%、同翅目个体数占各自捕获总个体数的45.8%～75.4%.杭州茶园中,纯白、湖蓝和天蓝色板诱捕同翅目种数小于各自捕获的总种数的20%、同翅目个体数小于各自捕获的总个体数的51%;其它7种色板诱捕同翅目种数大于各自捕获的总种数的22.7%,果绿和墨绿诱捕的同翅目个体数占各自诱捕的总个体数的48.6%和49.8%,桃红、素馨黄、土黄、芽绿和桔黄诱捕的同翅目个体数占各自诱捕的总个体数的55.5%～78.1%.绍兴茶园中,素馨黄和芽绿诱捕的粉虱、假眼小绿叶蝉和广翅蜡蝉分别占两种色板诱捕的总个体数的67.7%和50.1%;杭州茶园中,素馨黄和芽绿诱捕的这几种害虫总数分别占二者诱捕的总个体数的70.7% 和59.2%.(3) 捕获的主要是害虫,绍兴茶园中诱捕的害虫数是天敌的1.63倍,杭州茶园中诱捕的害虫数是天敌的2.23倍.(4)色板之间引诱的种数和个体数差异显著.素馨黄和芽绿诱捕的种数和个体数最多、被捕昆虫的群落多样性指数很小,对叶蝉、粉虱和蜡蝉的引诱力最强;土黄和桔黄诱捕的种数和个体数较多、多样性指数较小,对同翅目昆虫的引诱力较强;纯白色诱捕的种数和个体数居中、多样性指数最大,即诱捕的各种昆虫的个体数差别小;果绿、桃红、墨绿和湖蓝诱捕的种数和个体数明显较少、多样性指数也较小;天蓝色诱捕的种数和个体数最少.(5) 寄生蝇类偏嗜蓝色,缨小蜂类、茧蜂类和小蜂类偏嗜芽绿色.建议将素馨黄和芽绿色板与信息素组合,分别用于监测和诱捕叶蝉和粉虱.     

Rice Resistance to Planthoppers and Leafhoppers

For over 50 years, host-plant resistance has been regarded as an efficient method to reduce yield losses to rice caused by delphacid and cicadelid hoppers. Already a number of resistant rice varieties have been developed and deployed throughout Asia. To date, over 70 hopper resistance genes have been identified in rice; however, less than 10 genes have been deliberately introduced to commercial rice varieties. Currently, due to recent brown planthopper (Nilaparvata lugens [Stål]) and whitebacked planthopper (Sogatella furcifera [Horvath]) outbreaks occurring at an unprecedented scale, researchers are working toward a second generation of resistant varieties using newly identified gene loci and applying new molecular breeding methods. This paper reviews advances in the identification of resistance genes and QTLs against hoppers in rice. It collates all published information on resistance loci and QTLs against the major rice planthoppers and leafhoppers and presents information on gene locations, genetic markers, differential varieties, and wild rice species as sources of resistance. The review indicates that, whereas progress in the identification of genes has been rapid, considerable tidying of the information is required, especially regarding gene nomenclature and resistance spectra. Furthermore, sound information on gene functioning is almost completely lacking. However, hopper responses to resistance mechanisms are likely to be similar because a single phenotyping technique has been applied by most national and international breeding programs during germplasm screening. The review classifies genes occurring at two chromosome regions associated with several identified resistance loci and highlights these (Chr4S: BphR-R and Chr12L: BphR-R) as general stress response regions. The review calls for a greater diversity of phenotyping methods to enhance the durability of resistant varieties developed using marker-aided selection and emphasizes a need to anticipate the development of virulent hopper populations in response to the field deployment of genes.     

Probing and feeding characteristics of the greenhouse whitefly in association with host-plant acceptance and whitefly strains

Host-plant and whitefly strain effects and their interactions on the probing and sap feeding of the greenhouse whitefly, Trialeurodes vaporariorum (Westwood), have been investigated in this study using the DC-EPG (Electrical Penetration Graph) technique. Whiteflies generally displayed fewer but longer probes on highly acceptable cucumber than on less acceptable tomato. Both whitefly strains, the T(omato)-strain and the C(ucumber)-strain, showed a significantly lower number of phloem phases on cucumber than on tomato. However, the duration of total phloem phases achieved by either of the whitefly strains on these two host plants was not significantly different. These data indicate that a more continuous phloem feeding has occurred on cucumber plants. Indeed, the percentage of phloem feeding time after the first sustained phloem phase (longer than 15 min) was higher on cucumber for the C-strain whiteflies. When comparing these two whitefly strains, the T-strain whiteflies probed less frequently but longer than the C-strain whiteflies did on both host plants. Also, the T-strain whiteflies displayed a longer duration of total phloem phases on tomato. An interaction between the whitefly strain and plant effects was detected on a parameter, which showed that whiteflies probed significantly longer before reaching the first phloem phase on the host plants that had been previously experienced. In conclusion, both plant species and whitefly strains affect whitefly's probing and feeding behaviour, though plant effects are much stronger.     

茶树CsCCD1和CsCCD4基因的克隆和表达分析

该研究以‘铁观音’茶树为材料,同源克隆了茶树类胡萝卜素裂解双加氧酶基因CsCCD1和CsCCD4 (NCBI登录号分别为MH119136和MH119137)全长cDNA。序列分析结果显示,CsCCD1序列全长1 766 bp,包含1 641 bp开放阅读框(ORF),编码545个氨基酸,定位于细胞质中,不存在跨膜结构和信号肽;CsCCD4序列全长1 942 bp,包含1 842 bp ORF,编码612个氨基酸,不存在跨膜结构,但在N端具有叶绿体转运肽,定位于质体中。进化树分析显示,CsCCD1和CsCCD4与杜鹃聚为一类。荧光定量检测显示,CsCCD1和CsCCD4在叶、茎和花中表达量较高。在乌龙茶做青过程中,CsCCD1和CsCCD4基因都是从鲜叶到晒青表达量下调,而CsCCD1在一摇和二摇显著上调表达,在三摇后下降;CsCCD4只在一摇后上调表达,之后表达量迅速降低。推测CsCCD1和CsCCD4基因可能与乌龙茶做青香气品质形成关系密切。