烟草水浸提液对葡萄根瘤蚜侵染危害的控制效应

【目的】检疫害虫葡萄根瘤蚜Daktulosphaira vitifoliae已经在我国多个地区葡萄园发现为害, 直接影响葡萄生长; 前期实验发现烟草水浸提液可有效控制离体根上根瘤蚜的存活率, 本研究进一步开展盆栽验证, 探讨烟草水浸提液对葡萄根瘤蚜侵染的控制效应及葡萄植株生长抑制的缓解作用。【方法】以两年生温室盆栽‘巨峰’自根苗为实验材料, 接种根瘤蚜虫卵35 d后进行烟草水浸提液处理, 处理后测定供试植株的根结数、生长量、根系活力、叶片光合速率及叶绿素荧光等指标。【结果】20 mg/mL和50 mg/mL的烟草水浸提液对根瘤蚜均有一定程度的抑制作用, 施药21 d后, 葡萄根部的新增根结数分别比侵染对照下降了42.9%和52.8%。烟草水浸提液处理还在一定程度上缓解了根瘤蚜侵染对植株的光合能力和荧光的抑制作用。20 mg/mL和50 mg/mL烟草水浸提液处理的株高分别比侵染对照提高了57.1%和7.7%; 根瘤蚜侵染降低了土壤中真菌、细菌和放线菌的数量, 烟草水浸提液处理进一步降低了真菌和放线菌的数量, 降低了由微生物带来的次生侵染几率。【结论】烟草水浸提液对葡萄根瘤蚜的侵染具有控制效果, 并可对葡萄根瘤蚜侵染已造成的寄主危害有一定的缓解作用。     

两个葡萄杂交后代根系抗葡萄根瘤蚜及抗寒性鉴定

【目的】为了筛选抗葡萄根瘤蚜Daktulosphaira vitifoliae Fitch且抗寒的葡萄砧木以适应我国葡萄生产需求。【方法】以山葡萄Vitis amurensis Rupr.左山1号×SO4杂种F1代的45个株系(A系列)和左山1号×101-1杂种F1代27个株系(B系列)为试材,采用离体根接种鉴定法进行抗葡萄根瘤蚜鉴定及抗性分级;采用差热分析系统(differential thermal analysis,DTA)进行各株系根系的低温放热(low temperature exotherms,LTE)分析,建立各株系根系韧皮部及木质部的温度-伤害度(LT-I)回归方程,评估各株系根系的抗寒性。【结果】葡萄根瘤蚜在杂交株系根系上的产卵量均显著低于敏感品种巨峰,筛选出被葡萄根瘤蚜侵染后不能形成根瘤,抗葡萄根瘤蚜级别为0级的A系列杂交株系18个和B系列株系11个。被葡萄根瘤蚜侵染后形成根瘤比例低于10%的抗葡萄根瘤蚜级别为1级的A系列杂交株系9个和B系列株系4个;筛选出A系列综合低温放热温度隶属度函数、韧皮部和木质部低温放热温度隶属度函数3个指标均低于贝达的株系27个,B系列各指标均低于贝达的株系3个。【结论】本研究筛选出抗寒性强且对葡萄根瘤蚜抗性强的A系列株系15个和B系列株系2个。其中,A14,A16,A18,A22,A23,A28,A34,A35,A38,A44,A50,B24和B26对葡萄根瘤蚜抗性级别为0;A11,A15,A17和A27对葡萄根瘤蚜抗性级别为1。     

根瘤蚜侵染对不同抗性葡萄品种根系超微结构的影响

【目的】探讨不同抗性葡萄品种对葡萄根瘤蚜Daktulosphaira vitifolia Fitch侵染后的组织结构响应。【方法】以砧木140Ru (Vitis rupestris×V. berlandieri)和栽培品种赤霞珠Vitis vinifera cv. Cabernet Sauvignon离体三级根及140Ru盆栽苗为试材接种根瘤蚜卵, 接种两周后取刺吸位点组织作超薄切片并观察超微结构变化, 取盆栽140Ru不同时期根结测定多酚含量。【结果】超微结构显示, 砧木140Ru根系周皮层最外层细胞壁的厚度（1 031.25 nm）及周皮层细胞层数（6～7层）显著高于赤霞珠品种的周皮层细胞厚度（543.75 nm）及层数（3～4层）, 140Ru根系韧皮部酚类物质含量比赤霞珠高出35％; 侵染后, 砧木140Ru周皮层细胞第3层以内的细胞壁加厚并积聚大量的多酚类物质, 140Ru葡萄新根被根瘤蚜侵染后多酚含量呈升高趋势, 在接种20 d时是对照的2.4倍; 赤霞珠粗根被根瘤蚜侵染后薄壁细胞中细胞质变浓, 出现大量淀粉粒, 线粒体及内质网数量增多。【结论】砧木140Ru的周皮层组织结构较赤霞珠不利于根瘤蚜口针穿刺, 被根瘤蚜侵染后发生了不利于根瘤蚜侵染取食的变化。     

外源茉莉酸诱导的青杨生化抗性及其对舞毒蛾幼虫食物利用的影响

【目的】为探讨外源茉莉酸(jasmonic acid, JA)诱导的青杨 Populus cathayana Rehd.抗性对舞毒蛾 Lymantria dispar (L.)幼虫食物利用的影响。【方法】在室内对青杨苗木喷施0.001和0.1 mmol/L两种浓度的茉莉酸,对照喷0.2%的丙酮水溶液,喷施后1, 5和10 d分别采集叶片,分析其初生和次生物质含量的变化。另选喷施0.1 mmol/L茉莉酸的青杨苗木,喷施后1, 5和10 d分别接舞毒蛾2龄幼虫,单头饲养,测定其取食量、体重和排粪量及发育历期,统计分析其食物消化率、转化率和利用率。【结果】两种浓度茉莉酸处理均使青杨叶片中的蛋白质和可溶性糖含量降低,而木质素、单宁、黄酮和酚类物质含量增加。蛋白质和可溶性糖在处理第10天时含量最低,其中高浓度茉莉酸处理较对照分别降低46.5%和49.1%,低浓度处理分别降低30.6%和22.8%。叶片中酚类物质含量在高浓度处理第10 天时增幅最大,较对照增加102%。木质素、黄酮和单宁酸在处理第5天时的含量最高,其中高浓度处理分别较对照增加113%, 75%和57%。用0.1 mmol/L茉莉酸诱导处理后,舞毒蛾2龄幼虫对叶片的消化率、转化率和利用率均有所降低, 取食处理后1, 5和10 d的青杨叶片的食物利用率较对照分别降低了29.4%, 27.6%和28.2%,且幼虫体重降低、发育历期延长。【结论】结果提示外源茉莉酸诱导青杨可对舞毒蛾产生明显的生化抗性,实践中可利用这种生化抗性防治舞毒蛾的危害。     

钙肥对葡萄根瘤蚜生长发育和繁殖的影响

【目的】为明确葡萄所需的大量元素钙对葡萄根瘤蚜 Daktulosphaira vitifoliae(Fitch）生长发育和繁殖的影响。【方法】在温室中,设置不同 Ca⁽²⁺⁾浓度（0、2 和 4mmol·L(2+)浓度（0、2 和 4mmol·L^(-1)）的钙全营养液于葡萄全生育期浇灌葡萄,评价离体根段对葡萄根瘤蚜的存活率、若蚜、成蚜寿命和繁殖力的影响。【结果】随着钙离子浓度的增加,葡萄根瘤蚜在葡萄根上的 1 龄若蚜历期缩短 0.80 倍以上,成蚜历期增加超过 1.88倍,寿命显著延长超过 1.18 倍,产卵量提高 3.34 倍以上。与 0 mmol·L(-1)）的钙全营养液于葡萄全生育期浇灌葡萄,评价离体根段对葡萄根瘤蚜的存活率、若蚜、成蚜寿命和繁殖力的影响。【结果】随着钙离子浓度的增加,葡萄根瘤蚜在葡萄根上的 1 龄若蚜历期缩短 0.80 倍以上,成蚜历期增加超过 1.88倍,寿命显著延长超过 1.18 倍,产卵量提高 3.34 倍以上。与 0 mmol·L^(-1) Ca(-1) Ca⁽²⁺⁾和 2 mmol·L(2+)和 2 mmol·L^(-1) Ca(-1) Ca⁽²⁺⁾处理相比,葡萄根瘤蚜在 4 mmol·L(2+)处理相比,葡萄根瘤蚜在 4 mmol·L^(-1) Ca(-1) Ca⁽²⁺⁾处理下的葡萄根上,内禀增长率 r_m、周限增长率 λ 和净增殖率 R_0均最高,分别为（0.191 ± 0.003）、（1.210 ± 0.004）和（169.070 ± 11.897)(P < 0.05）,且世代发育历期显著缩短（P <0.05）。特定年龄龄期存活率 S_(xj)、种群特定年龄繁殖力 m_x和种群特定年龄繁殖值 l_xm_x在 4mmol·L(2+)处理下的葡萄根上,内禀增长率 r_m、周限增长率 λ 和净增殖率 R_0均最高,分别为（0.191 ± 0.003）、（1.210 ± 0.004）和（169.070 ± 11.897)(P < 0.05）,且世代发育历期显著缩短（P <0.05）。特定年龄龄期存活率 S_(xj)、种群特定年龄繁殖力 m_x和种群特定年龄繁殖值 l_xm_x在 4mmol·L^(-1)Ca(-1)Ca⁽²⁺⁾处理种群中保持较高水平,随着钙浓度降低,葡萄根瘤蚜的各生命参数明显降低。【结论】钙元素对葡萄根瘤蚜的生长发育和繁殖能力具有促进作用,葡萄根瘤蚜发生葡萄园更应合理使用钙肥。     

桑天牛卵啮小蜂雌蜂对茉莉酸诱导的桑枝挥发物的触角电位和嗅觉反应

【目的】明确桑天牛卵啮小蜂Aprostocetus prolixus雌蜂对茉莉酸诱导的桑Morus alba枝挥发物的触角电位和嗅觉反应,有利于揭示茉莉酸诱导桑枝产生间接抗性的化学机制。【方法】采用动态顶空气体法收集茉莉酸处理后的桑枝挥发物,分别通过气相色谱-触角电位联用仪(GC-EAD)和气相色谱/质谱联用仪(GC/MS)筛选和鉴定对桑天牛卵啮小蜂雌蜂有EAD活性的化合物,并比较这些活性组分的释放速率在茉莉酸处理桑枝与蒸馏水处理的对照桑枝间的差异,最后利用Y-型嗅觉测定仪明确对该寄生蜂有引诱活性的组分。【结果】桑枝经茉莉酸处理后,其挥发物中的Z-3-己烯醇、γ-萜品烯、β-水芹烯、紫苏烯、水杨酸甲酯及3种未知化合物使桑天牛卵啮小蜂雌蜂产生明显的EAD反应,并且β-水芹烯、紫苏烯和水杨酸甲酯的释放量明显高于对照,其中β-水芹烯和紫苏烯仅存在于茉莉酸处理后桑枝的挥发物中,而Z-3-己烯醇的释放速率与对照无显著差异。0.1%β-水芹烯、0.1%紫苏烯以及0.01%和0.001%水杨酸甲酯对该蜂雌蜂具有显著的引诱作用。【结论】结果提示,茉莉酸促进桑枝大量释放β-水芹烯、紫苏烯和水杨酸甲酯,是茉莉酸诱导桑树对桑天牛产生间接抗性的重要机制。     

根瘤蚜侵染不同抗性葡萄对根结形成及植株营养消耗的影响

为探讨不同抗性水平的葡萄砧木和品种被葡萄根瘤蚜Daktulosphaira vitifoliae Fitch侵染形成根结的状况及其对植株营养消耗的影响,采用盆栽方式接种根瘤蚜,测定了葡萄根结数量、质量及碳水化合物和脯氨酸含量等指标。结果表明：砧木5BB,1103P,SO4,3309C,101-14Mgt被根瘤蚜侵染后均不形成根结;140Ru,Lot和110R仅形成少量根结而不能形成根瘤;砧木贝达、栽培品种赤霞珠、巨峰和达米娜不仅能够形成根结和根瘤,而且根系腐烂程度严重。越敏感的品种形成的根结数量和体积越大,达米娜、巨峰和赤霞珠根结重占根重比例分别高达40.02％,37.08％和35.36％,百个根结质量分别是140Ru的4.4,3.5和433倍。3个形成根结的砧木的枝条生长量平均减少16.5％,而4个高感品种的生长量平均减少43％。根结质量与根结内的营养积聚呈正相关,其中根结质量与淀粉含量增加量的相关系数达0.94,与可溶性糖含量增加量的相关系数达0.81,与游离脯氨酸含量增加量的相关系数达0.93。     

部分葡萄品种和砧木抗葡萄根瘤蚜性能鉴定

为了检测我国葡萄品种和砧木对中国生态型葡萄根瘤蚜Daktulosphaira viticola (Fitch) 的抗性,在实验室条件下采用离体根接种法鉴定了10个葡萄砧木、18个葡萄栽培品种,测定了根瘤蚜存活率、发育历期、产卵量、种群龄期结构指数和成虫大小等指标。结果表明： 砧木品种SO4,5BB,1103P,420A,Lot,101-14MG,3309C,140Ru和110R抗根瘤蚜,接种后蚜虫陆续死亡,尤其是SO4,5BB,1103P和420A对根瘤蚜高抗,使根瘤蚜滞育仅发育至1龄。我国当作砧木使用的贝达不抗根瘤蚜,接种后蚜虫存活率始终保持在38.82％～47.5%,日产卵量达9.01;欧美杂交种中的康克、康拜尔和卡它巴对根瘤蚜稍有抗性,根瘤蚜日产卵量在6.00～6.85粒之间,其中康克、康拜尔葡萄上蚜虫的存活率随时间的延长而明显降低,由最初的60％下降至25％～30％; 欧美杂交种中的黑虎香和白香蕉虽然在一定程度上能够限制根瘤蚜的存活和发育进程,但对根瘤蚜的繁殖量没有影响。所有供试欧亚种和欧美杂交种的栽培品种都对根瘤蚜高度敏感,接种后第29 d 蚜虫存活率仍保持在40％～60％,且前后变化较小,日产卵量变化于7～12.35粒之间,总产卵量均在190粒以上。由此可见,我国供试栽培品种均不抗根瘤蚜,在根瘤蚜疫区需要用抗性砧木嫁接苗建园。     

外源化合物诱导后小麦对麦长管蚜和粘虫的抗虫性研究

喷施外源化合物和昆虫取食均可诱导小麦的防御反应,如挥发物组成发生改变、某些次生物质含量增加或减少,进而影响昆虫和天敌的行为反应。本实验中小麦苗经茉莉酸或几丁质喷施诱导,可显著提高小麦中丁布的含量,但茉莉酸甲酯的诱导作用不明显。喷施茉莉酸及麦长管蚜或粘虫取食诱导小麦,会导致小麦挥发物的种类及含量与对照相比有显著不同,3种处理间小麦的挥发物也存在显著差异。用经茉莉酸处理的麦苗饲养麦长管蚜和粘虫,可显著抑制二者的生长发育速度,使体重减轻,并导致麦长管蚜繁殖力下降。经茉莉酸处理的麦苗的挥发物对粘虫寄生蜂有吸引作用,表明茉莉酸在诱导小麦产生直接抗虫性的同时,还能诱导其释放吸引天敌的挥发性化合物从而产生间接抗性。     

外源茉莉酸诱导菜豆对西花蓟马和南方小花蝽的行为反应

【目的】为明确茉莉酸诱导的菜豆对西花蓟马Frankliniella occidentalis和南方小花蝽Orius similis的行为反应。【方法】采用四臂嗅觉仪测定了西花蓟马和南方小花蝽对不同浓度茉莉酸诱导菜豆后的行为反应,并用气相色谱-质谱联用仪测定了不同浓度茉莉酸处理后菜豆挥发物的成分。【结果】不同浓度茉莉酸处理的菜豆植株对西花蓟马和南方小花蝽分别有不同程度的驱避和吸引作用,以1 mmol/L的茉莉酸处理植株对西花蓟马的驱避作用最强,0.1 mmol/L的茉莉酸处理植株对南方小花蝽的吸引作用最强。不同处理菜豆的挥发物在含量和成分上存在显著差异,(Z)-3-己烯丙酸酯、2-异丙基-甲氧基毗嗦只有在茉莉酸处理植株中检测到。结合不同浓度茉莉酸处理植株对西花蓟马和南方小花蝽的行为反应及菜豆挥发物含量的变化趋势,推测(E)-2-己烯醛对西花蓟马有驱避作用,(E)-2-乙酸叶醇酯对南方小花蝽具有引诱作用。【结论】茉莉酸处理菜豆后,植物挥发物种类和含量发生了变化,在增强菜豆植株抗虫性的同时,还可增强捕食性天敌南方小花蝽的搜索和捕食能力。     

根瘤蚜对不同抗性葡萄的选择性及葡萄根系挥发性物质的鉴定

为了探讨不同抗性水平砧木及品种对根瘤蚜吸引及驱避效应以及鉴定挥发性物质组分,采用离体根段培养法测定葡萄根瘤蚜Phylloxera viticola Fitch对砧木5BB,140Ru及栽培品种巨峰的选择性,发现根瘤蚜对栽培品种巨峰的根段具有很强的选择性,至接种后3 d,67.13％的根瘤蚜优先选择巨峰;而对5BB和140Ru的选择性却很差,分别有15.29％和17.58％的根瘤蚜选择5BB及140Ru。根瘤蚜在不同品种根段上的生长发育和繁殖也存在明显差异。对砧木5BB和巨峰根系的挥发性组分进行测定,发现2个品种挥发物独有组分数量均较高,巨峰独有组分为23个,占总成分数的48.94％,5BB的独有组分为32个,占总成分数的57.14％。5BB和巨峰的共有组分为24个,主要成分均为亚油酸甲酯和反油酸甲酯,但含量有所不同,其中,5BB以倍半萜类的丁香醇及脂肪酸甲酯类的亚油酸甲酯,反油酸甲酯,棕榈油酸甲酯含量较高,高于巨峰中相应物质含量的4.23%~6.46％,而巨峰以邻苯二甲酸二丁酯及胆甾烷含量较高,分别高于5BB相应组分含量的6.81％和1.07％。结果说明葡萄根瘤蚜对不同抗性水平砧木及品种确实存在不同的选择性,并且不同抗性水平的砧木及品种的挥发性物质存在显著差异。     

Jasmonic acid induced resistance in grapevines to a root and leaf feeder

We investigated the effects of induced resistance to the folivore Pacific spider mite, Tetranychus pacificus McGregor (Acari: Tetranychidae), as well as the root-feeding grape phylloxera Daktulosphaira vitifoliae (Fitch) (Homoptera: Phylloxeridae) in grapevines using exogenous applications of the natural plant inducer, jasmonic acid. Foliar jasmonic acid application at concentrations that caused no phytotoxicity significantly reduced the performance of both herbivores. There were less than half as many eggs produced by spider mites feeding on the induced leaves compared with control grapevine leaves. Induction reduced the numbers of phylloxera eggs and nymphal instars by approximately threefold and twofold, respectively, on induced compared with control grapevine roots. The negative demographic effects of jasmonic acid application appeared to be caused by changes in fecundity for the Pacific spider mite, and possibly changes in development rate and fecundity for grape phylloxera.     

The Sublethal Effects of β-Ecdysterone,a Highly Active Compound from Achyranthes bidentata Blume,on Grape Phylloxera,Daktulosphaira vitifoliae Fitch

Grape phylloxera, Daktulosphaira vitifoliae (Fitch) (Hemiptera, Phylloxeridae), is a very destructive insect pest of grapevines. Intercropping of Achyranthes bidentata Blume (f. Amaranthaceae) and Vitis spp. grapevines can be useful to control this pest. In the present study, the toxicity of 22 compounds, known to be present in A. bidentata, to grape phylloxera was evaluated. All treatments were toxic towards grape phylloxera but the degree of toxicity differed between treatments. Among the 22 tested compounds, several of which proved toxic towards grape phylloxera. However β-ecdysterone had higher toxic effects against grape phylloxera, with LC₅₀ values of 175.73 mg a.i. liter^-1. In addition, we assessed the sublethal effects of LC₁₀, LC₂₀ and LC₄₀ of β-ecdysterone on grape phylloxera. The fourth instar and adult developmental periods and total life span were significantly prolonged by LC₄₀ of β-ecdysterone. Fecundity decreased when grape phylloxera were exposed to LC₂₀ and LC₄₀ of β-ecdysterone. In addition, LC₄₀ of β-ecdysterone decreased the intrinsic rate of increase (r_m) and the finite rate of increase (λ) and prolonged the population doubling time (DT). The net reproductive rate (R₀) was significantly reduced by both the LC₂₀ and LC₄₀ β-ecdysterone treatments. Our results demonstrated that β-ecdysterone had higher toxic effects and significant sublethal effects on grape phylloxera, and showed potential control of grape phylloxera.     

Seasonal capacity of attached and detached vineyard roots to support grape phylloxera (Homoptera: Phylloxeridae)

Field experiments were conducted to evaluate population densities and survival, developmental rate, and fecundity of grape phylloxera, Daktulosphaira vitifoliae (Fitch), as influenced by root attachment or detachment from mature, field-grown, Vitis vinifera L. grapevines through the growing season. Experiments were performed using artificial infestations of California biotype A grape phylloxera. Thirty-day bioassays on attached- and detached-roots were repeated monthly from May to September in 1997 (cultivar 'Carignane') and April to September in 1998 (cultivar 'Thompson Seedless'). The bioassays showed that attached roots had lower population densities than detached roots in all months of both years. Densities varied by month, tending to be higher in spring than in summer. Of the population parameters studied, survival was most influenced by attachment condition, being higher on detached than on attached roots by up to 25-fold. These results imply the importance of vine-related mortality factors to grape phylloxera population density. Influence of root attachment condition on developmental rate and fecundity was not uniform across bioassay months for either year; however, in the four out of 21 assays where there was a significant difference it favored detached roots by twofold. Fruit harvest resulted in higher survival in the July assay but not for assays in August and September; however, neither developmental rate nor fecundity was affected by harvest in any ofthe assays. We conclude that mortality rather than nutritional factors are most limiting for field populations on susceptible vines. This work suggests that detachment of roots as occurs with root girdling by root pathogens may increase grape phylloxera populations on infested, susceptible vines. These results imply that excised root bioassays over-estimate grape phylloxera virulence and underestimate rootstock resistance.     

南美斑潜蝇幼虫为害对黄瓜叶片中水杨酸和茉莉酸的诱导作用

【目的】为揭示南美斑潜蝇Liriomyza huidobrensis (Blanchard)与其寄主相互作用的机理, 为利用诱导抗性控制南美斑潜蝇的发生为害奠定必要的基础。【方法】本文采用高效液相色谱法（HPLC）和超高效液相色谱法-质谱联用法（UPLC MS）, 分别测定了南美斑潜蝇幼虫为害对黄瓜叶片中茉莉酸（jasmonic acid, JA）和水杨酸（salicylic acid, SA）的诱导作用。【结果】南美斑潜蝇幼虫持续为害1 d后, 受害黄瓜叶片内JA含量即显著高于健康对照, 轻度受害处理和重度受害处理分别在第3天和第5天上升幅度最大, 分别比健康对照增加2.01倍和1.62倍; 而SA含量在3 d后才显著高于健康对照, 轻度受害处理和重度受害处理在第9天上升幅度最大, 分别比健康对照增加4.66倍和1.67倍; 轻度受害对JA和SA的系统诱导作用不明显, 而重度受害对JA和SA具有明显的系统诱导作用。【结论】南美斑潜蝇幼虫为害对黄瓜叶片内JA和SA具有诱导作用。     

Effect of Jasmonic, Salicylic, and Abscisic Acids on [14C]Leucine Incorporation into Proteins of Pea Leaves

All investigated exogenous phytohormones (jasmonic, salicylic, and abscisic acids) induced the appearance of ¹⁴C-label in a polypeptide with molecular mass 29 kD that was not found in the control; these acids also increased [¹⁴C]leucine incorporation into a 25-kD polypeptide and decreased such incorporation into a 45-kD polypeptide. This can be considered as a nonspecific response of the plants to the action of these hormones. Salicylic and abscisic (but not jasmonic) acids induced the synthesis of a 19-kD polypeptide, and jasmonate induced the synthesis of a 96-kD polypeptide.