利用性信息素防治豆野螟的生物学基础及应用技术研究
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摘要
豆野螟Maruca vitrata是一种严重的泛热带豆类蔬菜害虫。主要危害豇豆Vigna unguiculata、四季豆Phaseolus vulgaris、扁豆Dolichos lablab、木豆Cajanus cajan等表面少毛的豆类蔬菜。对豇豆的危害尤为严重，此虫为害豇豆等常造成“十荚九蛀”，一般田块花被害率52.3％以上，豆荚被害率37.8％，叶被害率9.9％，大发生年份若不进行防治几乎绝收，花被害率可达到90％以上，产量损失达到20％～60％。幼虫钻蛀或结网为害花、花蕾、嫩茎、豆荚，严重影响化学农药的使用效果，同时也不利于天敌对其的自然控制作用。鉴于此，我们利用性信息素来作为监测和防治豆野螟的手段，主要结果如下：
1．豆野螟成虫行为学特征及性信息素产生与释放节律
豆野螟的羽化行为全天可见，在雌蛾中，86％于暗期羽化；在雄蛾中，73％于暗期羽化；雌雄蛾羽化行为在暗期第4、5和8 h差异达到显著。交尾活动发生在暗期19：00到5：00之间，交尾持续时间最短约为20 min，最长约为90 min，3日龄进入暗期第5 h具有最高的交尾率。1、6和7日龄成虫具有单个交尾高峰，2到5日龄成虫具有两个交尾高峰。同一日龄成虫交尾在暗期前半段平均花费的时间要明显高于在后半段花费的时间。低龄和高龄的成虫用于交尾的时间明显高于中龄的性成熟成虫。成虫的开始交尾时间随着日龄的增加逐渐前移。雄蛾对3日龄处女雌蛾进入暗期后第5和第9 h的性腺提取物和空气收集性信息素的触角电位反应最强。处女雌蛾田间诱蛾试验表明：23：00-01：00为诱蛾高峰期，3日龄处女雌蛾的诱蛾效果最好。该蛾的羽化、交尾及性信息素产生与释放均存在节律上的一致性。
2．豆野螟延迟交尾和多次交尾对其生殖活动的影响。
豆野螟雌雄同时延迟交尾，雌雄虫的寿命、产卵量均表现为先上升后下降的趋势，但是对卵的孵化率没有显著的影响；雌虫延迟交尾，随着延迟时间的增加，雌虫的寿命、产卵量、卵的孵化率表现为下降的趋势，而雄虫的寿命延迟交尾第三天达到最大值；雄虫延迟交尾，随着延迟交尾时间的增加，雌雄虫的寿命、产卵量均表现为先上升后下降的趋势，卵的孵化率随着延迟逐渐下降。随着雄虫交尾次数的增高，成功交尾率在逐渐降低，用于交尾的时间延长。雌虫和相应雄虫的寿命逐渐缩短，雌虫的产卵量下降，但对卵的孵化率影响不大。豆野螟雌虫一生只交尾一次，试验室未见到雌虫二次交尾。
3．豆野螟成虫日龄对交尾行为有较大的影响。
不同日龄雌蛾与3日龄雄蛾的交尾，交尾率随蛾龄增加呈下降趋势；不同日龄雄蛾与3日龄雌蛾交尾，交尾率随蛾龄增加呈升高趋势；相同日龄成蛾，交尾率先升后降。不同日龄雌蛾交尾百分率的下降在较高温环境比在较低温环境更加显著；但环境温度对不同日龄雄蛾的交尾却没有太大的影响；相同日龄成蛾3日龄以后的交尾在较高温环境下下降的更快。不同日龄雌蛾的交尾研究中，随着雌蛾日龄的增加，交尾持续时间明显延长；不同日龄雄蛾的交尾研究中，较低日龄和较高日龄雄蛾交尾时的持续时间较长。在较低温环境下所有试验各日龄的持续时间均有所延长。不同日龄雌蛾(雄蛾)与3日龄雄蛾(雌蛾)的交尾和相对应的雄蛾对雌蛾性信息素的触角电位反应(electroantennography，EAG)基本一致，说明雄蛾的反应在交尾行为的完成中具有重要的作用。
4．豆野螟性信息素组份化学结构的分析与鉴定
通过使用了DB-1和DB-WAX毛细柱进行气相色谱(gas chromatography，GC)分析以及使用DB-17和DB-WAX毛细柱进行气相色谱-质谱(gas chrornatography-mass spectrometry，GC-MS)分析豆野螟武汉和广州两个地理种群性信息素腺体组份的化学结构，结果表明：(E，E)-10，12-hexadecadienal是豆野螟两个地理种群性信息素的主要组份。同时首次利用GC和GC-MS发现(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal也是武汉种群性信息素的组份，豆野螟武汉种群性信息素腺体成分为(E，E)-10，12-hexadecadienal，(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal，其比例为51.4：4.1：44.5(峰面积之比)。广州种群性信息素中没有发现(E，E)-10，12-hexadecadienol存在，其性信息素由(E，E)-10，12-hexadecadienal和(E)-10-hexadecenal组成，其比例为86.5：13.5(峰面积之比)。
5．豆野螟雄蛾触角对性信息素的EAG反应
三种化合物的剂量-反应关系研究表明，不论是武汉种群还是广州种群，豆野螟雄虫触角对(E，E)-10，12-hexadecadienal均最为敏感，而对另外两种标准化合物次之。二元组份的研究表明，两个种群诱芯中只要有(E，E)-10，12-hexadecadienal，EAG反应均较强，由(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal组成的诱芯活性最差；在武汉，(E，E)-10，12-hexadecadiena：(E)-10-hexadecenal为5：5的组合比其他二元组份产生更大的EAG反应；在广州，(E，E)-10，12-hexadecadienal：(E，E)-10，12-hexadecadienol为5：5的组合显著高于其他所有二元组合。三元诱芯的研究发现，在武汉，(E，E)-10，12-hexadecadienal：(E，E)-10，12-hexadecadienol：(E)-10-hexadecenal为5：1：5时产生最大的EAG反应，在广州，三种化合物为5：5：1的组合产生的EAG反应更大。
6．豆野螟触角化感器的超微结构观察
豆野螟雌雄蛾触角呈线状，由柄节、梗节和鞭节组成。柄节较长；梗节较粗，并均覆盖着鳞片；鞭节由72～82节组成，长度为86～97 mm，两性间无明显的差别。豆野螟感受器主要分布在触角的背面、腹面和外侧面，内侧面一般覆盖有鳞片，因此感受器很少。雌雄比较发现，就长度而言，雌蛾触角上的腔锥感器以及耳形感器均长于雄蛾，但是毛形感器则正好相反；就分布密度而言，三种感器在雌蛾触角上的分布比雄蛾较稀疏。两个地理种群的比较发现，广州种群不论雌雄蛾，其触角感器比武汉种群密度大，但长度偏短；刺形感器仅见于武汉种群的触角上，而锥形感器仅见于广州种群。
7．性信息素活性组份的田间诱捕试验
田间试验表明，在武汉，单一组份诱芯(E，E)-10，12-hexadecadienal的诱捕活性最佳，(E)-10-hexadecenal可以起到增效作用，而(E，E)-10，12-hexadecadienol没有增效作用；二元组份诱芯(E，E)-10，12-hexadecadienal和(E)-10-hexadecenal为5：5以及三元组份诱芯(E，E)-10，12-hexadecadienal、(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal的比例为5：0.3：5时具有最佳的诱捕活性。而在广州，单一组份诱芯(E，E)-10，12-hexadecadienal的诱捕活性最佳，(E，E)-10，12-hexadecadienol可以起到增效作用，而(E)-10-hexadecenal没有增效作用；相应的二元组合活性最高为(E，E)-10，12-hexadecadienal和(E，E)-10，12-hexadecadienol为5：5的比例；添加微量的(E)-10-hexadecenal有增效作用，(E，E)-10，12-hexadecadienal、(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal的三者的最佳比例为5：5：0.7。
8．豆野螟性信息素诱芯及诱捕器优化试验
诱芯与诱捕器的最优化试验表明，随着诱捕器在田间放置时间的延长，其引诱活性呈现逐渐降低，前2周的诱捕效果最好，以后逐渐下降。雄虫的捕获数量不受诱芯的包裹与否的影响，使用铝箔保护的诱芯和不使用其保护的诱芯的诱捕量间没有明显的差异。120cm是最佳的诱捕高度，90和150 cm的高度次之，30和180cm最差。传统的三角形粘胶诱捕器与5L的水瓶诱捕器诱蛾量相当，水盆诱捕器和1.5L诱捕器较差。100μg的诱芯含量诱捕效果较好，与300μg间没有显著差异，继续增加每个诱芯的总含量不但没有增加诱捕活性，相反，诱捕量有所下降。
The legume pod borer, Maruca vitrata, is a serious pantropical insect pest of grainlegumes, such as cowpea, Vigna unguiculata, common bean, Phaseolus vulgaris, hyacinthbean, Dolichos lablab and pigeon-pea, Cajanus cajan. In cowpea, infestation was higheron flowers (52.3%) than on pods (37.8%) and leaves (9.9%). Without control measures,flower infestation rates up to 90% were reported. The losses in grain yield have beenestimated to range from 20 to 60%. The larvae of M.vitrata damage flowers, flower buds,terminal shoots and young pods by webbing or boring, which protects the larvae fromnatural enemies and insecticides. The major results are summarized as following:
1. Study on adult behavior and circadian rhythm of sex pheromone production andrelease of M. vitrata
The results showed that the emergence activity of M. vitrata occurred throughout theday, and 86% of females and 73% of males emerged during the scotophase. Thedifferences of emergence behavior between both sexes were statistically significantduring the 4th, 5th and 8th hour of the scotophase (t＞4; P＜0.05). The mating behaviortook placed from 19:00 to 05:00. The mating duration varied from 20 to 90 minutes. Thehighest mating frequency occurred during the 5th hour of the scotophase for 3-day-oldmoths. Single mating peaks were observed for 1-, 6- and 7-day-old moths, while doublemating peaks were observed for 2- to 5-day-old moths. Adults of the same age spentlonger time in mating during the first half of the scotophase than during the second half ofthe scotophase. The mating behaviour of the moth was age-dependent. Young and oldpairs spent more time in mating than mature pairs of the middle age. Advanced onset oftime and peak for mating were observed directly by older moths. The EAG response ofmale moths was highest to the crude extract of pheromonal glands and air collection from3-day-old virgin female moths and during 5th and 9th hour of the scotophase. In the fieldtests, the highest percentage of the male moths was trapped during 23:00-01:00, and moremale moths were trapped by 3-day-old virgin female moths than by female moths of otherage. The emergence activity, mating activity and the female sex pheromone release weresynchronized at different ages and time of the scotophase. The sex pheromone release was almost constant throughout the scotophase, but mating behavior concentrated between thetwo mating peaks.
2. Effect of delayed mating and multiple mating of M. vitrata on reproduction
Delayed mating and multiple mating of M. vitrata affected the reproduction. Thelongevity of females and males, fecundity of females increased firstly, and then reduced,but fertility of eggs did not change much when males and females mating were delayed atsame time.; The longevity of females, fecundity of females and fertility of eggs reduced,but the longevity of males increased firstly, and then reduced when females mating weredelayed; The longevity of females and males and fecundity of females increased firstly,and then reduced, but fertility of eggs reduced when males mating were delayed; Themating success rate, the longevity of males and females, fecundity of females reduced,but mating duration was prolonged, fertility of eggs did not change much when maleswere multiple mating. Males can mate for 4 times in the whole life while females canmate only one time.
3. The age of M. vitrata affected the mating.
Effect of male and female age on the mating success and circadian rhythms of maleresponse to female sex pheromones of the legume pod borer were investigated. Theactivity of thc females remained at a high level until the 3rd days after adult emergence,whereas in males, the peak of activity occurred on the 3rd day and activity remained at ahigh level until the 6th days. The mating frequency of same-aged pair was low at1-day-old, reached a maximum at 3-day-old and thereafter decreased rapidly with age.The declines in female mating success were more pronounced under warm than coolthermocycles, while were not obvious for males. The time spend mating was longer inolder than in younger females. Very young and very old males generally spent more timein mating than mature individuals. The duration of mating was considerably prolonged atall ages under cool than warm thermocycles. Effect of male and female age on the matingsuccess and circadian rhythms of male response to female sex pheromones of the legumepod borer were synchronized, which showed male response play an important role incopulation of Maruca vitrata.
4. Identification of female sex pheromone of M. vitrata
Sex pheromone gland extractions from Maruca vitrata virgin female mothsoriginating in Wuhan and Guangzhou were analyzed by GC (DB-1, DB-WAX) andGC-MS (DB-17, DB-WAX). The result confirmed previously published finding that (E,E)-10, 12-hexadecadienal is the most abundant component of the legume pod borer. InWuhan, exactions were also found to contain (E, E)-10, 12-hexadecadienol and (E)-10-hexadecenal as components. The ratio of three chemicals is 51.4: 4.1:44.5 in thesex pheromone glands. In Guangzhou, (E, E)-10, 12-hexadecadienol can not be found insex pheromone gland extractions of Maruca vitrata. The ratio of two chemicals is 86.5:13.5 for (E, E)-10, 12-hexadecadienal and (E)-10-hexadecenal respectively.
5. Male EAG response to female sex pheromone
Dose-response curve of these chemicals demonstrated that (E, E)-10,12-hexadecadienal elicited the largest male EAG responses to either Wuhan orGuangzhou populaion, followed by (E, E)-10,12-hexadecadienol and (E)-10-hexadecenal.Study on binary synthetic lures showed greater EAG responses were to those including (E,E)-10,12-hexadecadienal, but two-component blends including (E,E)-10,12-hexadecadienol and (E)-10-hexadecenal elicited lowly males EAG responses. InWuhan, a two-components blend of (E, E)-10, 12-hexadecadienal and (E)-10-hexadecenalin a 5:5 ratio elicited significantly males EAG responses than any other binary syntheticlures; In Guangzhou, a two-components blend of (E, E)-10, 12-hexadecadienal and (E,E)-10, 12-hexadecadienol in a 5:5 ratio elicited significantly males EAG responses thanany other binary synthetic lures; Study on ternary synthetic lures showedthree-component blend of (E, E)-10, 12-hexadecadienal and two other components in a5:1:5 ratio elicited significantly males EAG responses in Wuhan. A three-componentblend of (E, E)-10, 12-hexadecadienal and two other components in a 5:5:1 ratio elicitedsignificantly males EAG responses in Guangzhou.
6. Observation on ultrastructure of antennal sensilla in M. vitrata
The morphology of antennae of M. vitrata was observed under scanning electronmicroscope. Antennae is made up of scapus, pedicle and flagella, which cosists of 72-82segments and is 86-97 cm long. The inner side surface of antenna is covered withcataphyUa and most of the antennal sensilla lie on its outer, upper and lower surfaces.Compared with both sesxes, Sensillum coeloconicum and Ear-shaped sensillum offemales are longer than that of males, but Sensillun trichodeum is shorter than that ofmales; The number of all Sensillum of females are lesser on females antennae than malesantennae. Compared with different population, the sensillum of Guangzhou populationare more compact and short for both sexes. Sensilla cheatica are observed on Wuhanpophlation antennae and sensilla basiconica can be observed on Guangzhou populationantennae.
7. Field-trapping experiment of synthetic sex pheromone lures
Field test with single synthetic sex pheromone lures demonstrated that (E, E)-10,12-hexadecadienal can catch more M. vitrata male in Wuhan, followed by (E, E)-10,12-hexadecadienol and (E)-10-hexadecenal. When adding (E)-10-hexadecenal to (E,E)-10, 12-hexadecadienal, the catches of males were increased significantly. Atwo-components lures of (E, E)-10, 12-hexadecadienal and (E)-10-hexadecenal in a 5:5ratio catched significantly male catches than any other binary synthetic lures; Athree-component lures of (E, E)-10, 12-hexadecadienal and two other components in a5:0.3:5 ratio catched significantly males than other ternary lures. In Guangzhou, (E, E)-10,12-hexadecadienal can catch more M. vitrata male, followed by (E,E)-10,12-hexadecadienol and (E)-10-hexadecenal. Adding (E, E)-10,12-hexadecadienolto (E, E)-10, 12-hexadecadienal, the catches of males were increased significantly. Atwo-components lures of (E, E)-10, 12-hexadecadienal and (E, E)-10, 12-hexadecadienolin a 5:5 ratio catched significantly males than any other binary synthetic lures; A ternarylures of (E, E)-10, 12-hexadecadienaI and two other components in a 5:5:0.7 ratio catchedsignificantly males.
8. Optimation of sex pheromone lure and trap for M. vitrata
The experiments were conducted to compare different trap designs, and other aspectsof the lures. Lures showed no loss of effectiveness for up to 2 weeks. Attraction reducedgradually 2 weeks later. Result showed that shielding the lures from the sunsight withaluminium foil did not increase catches of M. vitrata, and the data are not significantdifference with no-sielding lures. 120 cm was the optimum height for captures than 90 cmand 150 cm, and 30 cm and 180 cm were not feasible for capture. A sticky-trap design and5 L water-bottle trap were found to be superior to water-pan trap and 1.5 L water-bottle.100μg is the optimum lure dose for capture, and more dose will not increase the catches,in reverse, the dose inhabited the attractant.
1．豆野螟成虫行为学特征及性信息素产生与释放节律
豆野螟的羽化行为全天可见，在雌蛾中，86％于暗期羽化；在雄蛾中，73％于暗期羽化；雌雄蛾羽化行为在暗期第4、5和8 h差异达到显著。交尾活动发生在暗期19：00到5：00之间，交尾持续时间最短约为20 min，最长约为90 min，3日龄进入暗期第5 h具有最高的交尾率。1、6和7日龄成虫具有单个交尾高峰，2到5日龄成虫具有两个交尾高峰。同一日龄成虫交尾在暗期前半段平均花费的时间要明显高于在后半段花费的时间。低龄和高龄的成虫用于交尾的时间明显高于中龄的性成熟成虫。成虫的开始交尾时间随着日龄的增加逐渐前移。雄蛾对3日龄处女雌蛾进入暗期后第5和第9 h的性腺提取物和空气收集性信息素的触角电位反应最强。处女雌蛾田间诱蛾试验表明：23：00-01：00为诱蛾高峰期，3日龄处女雌蛾的诱蛾效果最好。该蛾的羽化、交尾及性信息素产生与释放均存在节律上的一致性。
2．豆野螟延迟交尾和多次交尾对其生殖活动的影响。
豆野螟雌雄同时延迟交尾，雌雄虫的寿命、产卵量均表现为先上升后下降的趋势，但是对卵的孵化率没有显著的影响；雌虫延迟交尾，随着延迟时间的增加，雌虫的寿命、产卵量、卵的孵化率表现为下降的趋势，而雄虫的寿命延迟交尾第三天达到最大值；雄虫延迟交尾，随着延迟交尾时间的增加，雌雄虫的寿命、产卵量均表现为先上升后下降的趋势，卵的孵化率随着延迟逐渐下降。随着雄虫交尾次数的增高，成功交尾率在逐渐降低，用于交尾的时间延长。雌虫和相应雄虫的寿命逐渐缩短，雌虫的产卵量下降，但对卵的孵化率影响不大。豆野螟雌虫一生只交尾一次，试验室未见到雌虫二次交尾。
3．豆野螟成虫日龄对交尾行为有较大的影响。
不同日龄雌蛾与3日龄雄蛾的交尾，交尾率随蛾龄增加呈下降趋势；不同日龄雄蛾与3日龄雌蛾交尾，交尾率随蛾龄增加呈升高趋势；相同日龄成蛾，交尾率先升后降。不同日龄雌蛾交尾百分率的下降在较高温环境比在较低温环境更加显著；但环境温度对不同日龄雄蛾的交尾却没有太大的影响；相同日龄成蛾3日龄以后的交尾在较高温环境下下降的更快。不同日龄雌蛾的交尾研究中，随着雌蛾日龄的增加，交尾持续时间明显延长；不同日龄雄蛾的交尾研究中，较低日龄和较高日龄雄蛾交尾时的持续时间较长。在较低温环境下所有试验各日龄的持续时间均有所延长。不同日龄雌蛾(雄蛾)与3日龄雄蛾(雌蛾)的交尾和相对应的雄蛾对雌蛾性信息素的触角电位反应(electroantennography，EAG)基本一致，说明雄蛾的反应在交尾行为的完成中具有重要的作用。
4．豆野螟性信息素组份化学结构的分析与鉴定
通过使用了DB-1和DB-WAX毛细柱进行气相色谱(gas chromatography，GC)分析以及使用DB-17和DB-WAX毛细柱进行气相色谱-质谱(gas chrornatography-mass spectrometry，GC-MS)分析豆野螟武汉和广州两个地理种群性信息素腺体组份的化学结构，结果表明：(E，E)-10，12-hexadecadienal是豆野螟两个地理种群性信息素的主要组份。同时首次利用GC和GC-MS发现(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal也是武汉种群性信息素的组份，豆野螟武汉种群性信息素腺体成分为(E，E)-10，12-hexadecadienal，(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal，其比例为51.4：4.1：44.5(峰面积之比)。广州种群性信息素中没有发现(E，E)-10，12-hexadecadienol存在，其性信息素由(E，E)-10，12-hexadecadienal和(E)-10-hexadecenal组成，其比例为86.5：13.5(峰面积之比)。
5．豆野螟雄蛾触角对性信息素的EAG反应
三种化合物的剂量-反应关系研究表明，不论是武汉种群还是广州种群，豆野螟雄虫触角对(E，E)-10，12-hexadecadienal均最为敏感，而对另外两种标准化合物次之。二元组份的研究表明，两个种群诱芯中只要有(E，E)-10，12-hexadecadienal，EAG反应均较强，由(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal组成的诱芯活性最差；在武汉，(E，E)-10，12-hexadecadiena：(E)-10-hexadecenal为5：5的组合比其他二元组份产生更大的EAG反应；在广州，(E，E)-10，12-hexadecadienal：(E，E)-10，12-hexadecadienol为5：5的组合显著高于其他所有二元组合。三元诱芯的研究发现，在武汉，(E，E)-10，12-hexadecadienal：(E，E)-10，12-hexadecadienol：(E)-10-hexadecenal为5：1：5时产生最大的EAG反应，在广州，三种化合物为5：5：1的组合产生的EAG反应更大。
6．豆野螟触角化感器的超微结构观察
豆野螟雌雄蛾触角呈线状，由柄节、梗节和鞭节组成。柄节较长；梗节较粗，并均覆盖着鳞片；鞭节由72～82节组成，长度为86～97 mm，两性间无明显的差别。豆野螟感受器主要分布在触角的背面、腹面和外侧面，内侧面一般覆盖有鳞片，因此感受器很少。雌雄比较发现，就长度而言，雌蛾触角上的腔锥感器以及耳形感器均长于雄蛾，但是毛形感器则正好相反；就分布密度而言，三种感器在雌蛾触角上的分布比雄蛾较稀疏。两个地理种群的比较发现，广州种群不论雌雄蛾，其触角感器比武汉种群密度大，但长度偏短；刺形感器仅见于武汉种群的触角上，而锥形感器仅见于广州种群。
7．性信息素活性组份的田间诱捕试验
田间试验表明，在武汉，单一组份诱芯(E，E)-10，12-hexadecadienal的诱捕活性最佳，(E)-10-hexadecenal可以起到增效作用，而(E，E)-10，12-hexadecadienol没有增效作用；二元组份诱芯(E，E)-10，12-hexadecadienal和(E)-10-hexadecenal为5：5以及三元组份诱芯(E，E)-10，12-hexadecadienal、(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal的比例为5：0.3：5时具有最佳的诱捕活性。而在广州，单一组份诱芯(E，E)-10，12-hexadecadienal的诱捕活性最佳，(E，E)-10，12-hexadecadienol可以起到增效作用，而(E)-10-hexadecenal没有增效作用；相应的二元组合活性最高为(E，E)-10，12-hexadecadienal和(E，E)-10，12-hexadecadienol为5：5的比例；添加微量的(E)-10-hexadecenal有增效作用，(E，E)-10，12-hexadecadienal、(E，E)-10，12-hexadecadienol和(E)-10-hexadecenal的三者的最佳比例为5：5：0.7。
8．豆野螟性信息素诱芯及诱捕器优化试验
诱芯与诱捕器的最优化试验表明，随着诱捕器在田间放置时间的延长，其引诱活性呈现逐渐降低，前2周的诱捕效果最好，以后逐渐下降。雄虫的捕获数量不受诱芯的包裹与否的影响，使用铝箔保护的诱芯和不使用其保护的诱芯的诱捕量间没有明显的差异。120cm是最佳的诱捕高度，90和150 cm的高度次之，30和180cm最差。传统的三角形粘胶诱捕器与5L的水瓶诱捕器诱蛾量相当，水盆诱捕器和1.5L诱捕器较差。100μg的诱芯含量诱捕效果较好，与300μg间没有显著差异，继续增加每个诱芯的总含量不但没有增加诱捕活性，相反，诱捕量有所下降。
The legume pod borer, Maruca vitrata, is a serious pantropical insect pest of grainlegumes, such as cowpea, Vigna unguiculata, common bean, Phaseolus vulgaris, hyacinthbean, Dolichos lablab and pigeon-pea, Cajanus cajan. In cowpea, infestation was higheron flowers (52.3%) than on pods (37.8%) and leaves (9.9%). Without control measures,flower infestation rates up to 90% were reported. The losses in grain yield have beenestimated to range from 20 to 60%. The larvae of M.vitrata damage flowers, flower buds,terminal shoots and young pods by webbing or boring, which protects the larvae fromnatural enemies and insecticides. The major results are summarized as following:
1. Study on adult behavior and circadian rhythm of sex pheromone production andrelease of M. vitrata
The results showed that the emergence activity of M. vitrata occurred throughout theday, and 86% of females and 73% of males emerged during the scotophase. Thedifferences of emergence behavior between both sexes were statistically significantduring the 4th, 5th and 8th hour of the scotophase (t＞4; P＜0.05). The mating behaviortook placed from 19:00 to 05:00. The mating duration varied from 20 to 90 minutes. Thehighest mating frequency occurred during the 5th hour of the scotophase for 3-day-oldmoths. Single mating peaks were observed for 1-, 6- and 7-day-old moths, while doublemating peaks were observed for 2- to 5-day-old moths. Adults of the same age spentlonger time in mating during the first half of the scotophase than during the second half ofthe scotophase. The mating behaviour of the moth was age-dependent. Young and oldpairs spent more time in mating than mature pairs of the middle age. Advanced onset oftime and peak for mating were observed directly by older moths. The EAG response ofmale moths was highest to the crude extract of pheromonal glands and air collection from3-day-old virgin female moths and during 5th and 9th hour of the scotophase. In the fieldtests, the highest percentage of the male moths was trapped during 23:00-01:00, and moremale moths were trapped by 3-day-old virgin female moths than by female moths of otherage. The emergence activity, mating activity and the female sex pheromone release weresynchronized at different ages and time of the scotophase. The sex pheromone release was almost constant throughout the scotophase, but mating behavior concentrated between thetwo mating peaks.
2. Effect of delayed mating and multiple mating of M. vitrata on reproduction
Delayed mating and multiple mating of M. vitrata affected the reproduction. Thelongevity of females and males, fecundity of females increased firstly, and then reduced,but fertility of eggs did not change much when males and females mating were delayed atsame time.; The longevity of females, fecundity of females and fertility of eggs reduced,but the longevity of males increased firstly, and then reduced when females mating weredelayed; The longevity of females and males and fecundity of females increased firstly,and then reduced, but fertility of eggs reduced when males mating were delayed; Themating success rate, the longevity of males and females, fecundity of females reduced,but mating duration was prolonged, fertility of eggs did not change much when maleswere multiple mating. Males can mate for 4 times in the whole life while females canmate only one time.
3. The age of M. vitrata affected the mating.
Effect of male and female age on the mating success and circadian rhythms of maleresponse to female sex pheromones of the legume pod borer were investigated. Theactivity of thc females remained at a high level until the 3rd days after adult emergence,whereas in males, the peak of activity occurred on the 3rd day and activity remained at ahigh level until the 6th days. The mating frequency of same-aged pair was low at1-day-old, reached a maximum at 3-day-old and thereafter decreased rapidly with age.The declines in female mating success were more pronounced under warm than coolthermocycles, while were not obvious for males. The time spend mating was longer inolder than in younger females. Very young and very old males generally spent more timein mating than mature individuals. The duration of mating was considerably prolonged atall ages under cool than warm thermocycles. Effect of male and female age on the matingsuccess and circadian rhythms of male response to female sex pheromones of the legumepod borer were synchronized, which showed male response play an important role incopulation of Maruca vitrata.
4. Identification of female sex pheromone of M. vitrata
Sex pheromone gland extractions from Maruca vitrata virgin female mothsoriginating in Wuhan and Guangzhou were analyzed by GC (DB-1, DB-WAX) andGC-MS (DB-17, DB-WAX). The result confirmed previously published finding that (E,E)-10, 12-hexadecadienal is the most abundant component of the legume pod borer. InWuhan, exactions were also found to contain (E, E)-10, 12-hexadecadienol and (E)-10-hexadecenal as components. The ratio of three chemicals is 51.4: 4.1:44.5 in thesex pheromone glands. In Guangzhou, (E, E)-10, 12-hexadecadienol can not be found insex pheromone gland extractions of Maruca vitrata. The ratio of two chemicals is 86.5:13.5 for (E, E)-10, 12-hexadecadienal and (E)-10-hexadecenal respectively.
5. Male EAG response to female sex pheromone
Dose-response curve of these chemicals demonstrated that (E, E)-10,12-hexadecadienal elicited the largest male EAG responses to either Wuhan orGuangzhou populaion, followed by (E, E)-10,12-hexadecadienol and (E)-10-hexadecenal.Study on binary synthetic lures showed greater EAG responses were to those including (E,E)-10,12-hexadecadienal, but two-component blends including (E,E)-10,12-hexadecadienol and (E)-10-hexadecenal elicited lowly males EAG responses. InWuhan, a two-components blend of (E, E)-10, 12-hexadecadienal and (E)-10-hexadecenalin a 5:5 ratio elicited significantly males EAG responses than any other binary syntheticlures; In Guangzhou, a two-components blend of (E, E)-10, 12-hexadecadienal and (E,E)-10, 12-hexadecadienol in a 5:5 ratio elicited significantly males EAG responses thanany other binary synthetic lures; Study on ternary synthetic lures showedthree-component blend of (E, E)-10, 12-hexadecadienal and two other components in a5:1:5 ratio elicited significantly males EAG responses in Wuhan. A three-componentblend of (E, E)-10, 12-hexadecadienal and two other components in a 5:5:1 ratio elicitedsignificantly males EAG responses in Guangzhou.
6. Observation on ultrastructure of antennal sensilla in M. vitrata
The morphology of antennae of M. vitrata was observed under scanning electronmicroscope. Antennae is made up of scapus, pedicle and flagella, which cosists of 72-82segments and is 86-97 cm long. The inner side surface of antenna is covered withcataphyUa and most of the antennal sensilla lie on its outer, upper and lower surfaces.Compared with both sesxes, Sensillum coeloconicum and Ear-shaped sensillum offemales are longer than that of males, but Sensillun trichodeum is shorter than that ofmales; The number of all Sensillum of females are lesser on females antennae than malesantennae. Compared with different population, the sensillum of Guangzhou populationare more compact and short for both sexes. Sensilla cheatica are observed on Wuhanpophlation antennae and sensilla basiconica can be observed on Guangzhou populationantennae.
7. Field-trapping experiment of synthetic sex pheromone lures
Field test with single synthetic sex pheromone lures demonstrated that (E, E)-10,12-hexadecadienal can catch more M. vitrata male in Wuhan, followed by (E, E)-10,12-hexadecadienol and (E)-10-hexadecenal. When adding (E)-10-hexadecenal to (E,E)-10, 12-hexadecadienal, the catches of males were increased significantly. Atwo-components lures of (E, E)-10, 12-hexadecadienal and (E)-10-hexadecenal in a 5:5ratio catched significantly male catches than any other binary synthetic lures; Athree-component lures of (E, E)-10, 12-hexadecadienal and two other components in a5:0.3:5 ratio catched significantly males than other ternary lures. In Guangzhou, (E, E)-10,12-hexadecadienal can catch more M. vitrata male, followed by (E,E)-10,12-hexadecadienol and (E)-10-hexadecenal. Adding (E, E)-10,12-hexadecadienolto (E, E)-10, 12-hexadecadienal, the catches of males were increased significantly. Atwo-components lures of (E, E)-10, 12-hexadecadienal and (E, E)-10, 12-hexadecadienolin a 5:5 ratio catched significantly males than any other binary synthetic lures; A ternarylures of (E, E)-10, 12-hexadecadienaI and two other components in a 5:5:0.7 ratio catchedsignificantly males.
8. Optimation of sex pheromone lure and trap for M. vitrata
The experiments were conducted to compare different trap designs, and other aspectsof the lures. Lures showed no loss of effectiveness for up to 2 weeks. Attraction reducedgradually 2 weeks later. Result showed that shielding the lures from the sunsight withaluminium foil did not increase catches of M. vitrata, and the data are not significantdifference with no-sielding lures. 120 cm was the optimum height for captures than 90 cmand 150 cm, and 30 cm and 180 cm were not feasible for capture. A sticky-trap design and5 L water-bottle trap were found to be superior to water-pan trap and 1.5 L water-bottle.100μg is the optimum lure dose for capture, and more dose will not increase the catches,in reverse, the dose inhabited the attractant.
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