体外培育赤眼蜂研究进展

体外培育赤眼蜂的国内外研究已达到相当水平:随着人工饲料配方的优化,选用的人工卵壳材料更适合于赤眼蜂的寄生与发育,以及人工卵卡机的研制成功和性能不断改进,产业化体外培育赤眼蜂已成为可能.体外培育赤眼蜂的品质管理问题是实现产业化体外培育赤眼蜂的关键.然而,国内外对体外培育赤眼蜂的品质管理研究涉及甚少,有待进一步研究.本文指出应开展体外培育赤眼蜂生物学特性、行为学特性和分子生物学特性的研究,并应用于品质管理,建立体外培育赤眼蜂的品质管理体系,促进实现体外培育赤眼蜂产业化的进程,从而满足农业生产上对赤眼蜂种群的大量需要.另外,人工卵滞育贮存法的突破有助于实现体外培育赤眼蜂的产业化,也应加强研究.最后,本文对体外培育赤眼蜂的应用前景进行了展望.     

以无昆虫物质的人工饲料培育松毛虫赤眼蜂幼虫

本文报道用不含昆虫物质的培养液以及另加鸡胚提取液、牛奶、鸡蛋黄、牛血清、酵母提取液、麦芽制品、蛋白胨配制成的人工饲料,以悬滴培养法在寄主卵外培育松毛虫赤眼蜂(Trichogramma dendrolimi)的结果。在最佳配方的人工饲料中,这种赤眼蜂可以从卵发育到成虫,蛹化率为36—53%,羽化率为7—16%,展翅率为12—34%。蜂体大小和发育速度与用柞蚕(Antheraea pernyi)卵培育的相似,但生活力略差。成蜂有交配能力,并可在柞蚕卵内产卵,后代以雌性个体为多。 用减缩法明确了培养液A及B组的不同部分对赤眼蜂幼虫发育的影响,证明了鸡蛋黄和培养液A在培育中有较重要的作用。但产卵的试验表明仅培养液A能吸引雌蜂产卵,而培养液B的不同成分引诱产卵的效果较差。     

小菜蛾卵作为赤眼蜂中间寄主的寄生适合度分析

【目的】寻求快速扩繁赤眼蜂种群的寄主,试以繁殖速率较高的小菜蛾为中间寄主,评价不同赤眼蜂种类对小菜蛾卵的寄生适合度并筛选出适合的蜂种。【方法】通过构建稻螟赤眼蜂Trichogramma japonicum Ashmead(简称T.j)、螟黄赤眼蜂T chilonis Ishi(简称T.c)和斑螟分索赤眼蜂Trichogrammatoidea hypsipylae Nagaraja(简称T.h)3种赤眼蜂在小菜蛾Plutella xylostella L.卵上繁殖的实验种群生命表,分析比较其各自的生命表参数,以了解不同蜂种对小菜蛾卵的寄生适合度。【结果】在小菜蛾卵上繁殖的稻螟赤眼蜂的净生殖力R_0、内禀增长率r_m、周限增长率λ与平均世代历期T分别为4.66、0.1633、1.1809、9.2532;斑螟分索赤眼蜂的上述4项参数分别为9.10、0.2177、1.2432、10.1432;螟黄赤眼蜂对小菜蛾卵的寄生能力较差,内禀增长率r_m为0.0338,且其后代性比偏雄,雌雄性比为1♀:6.2(?),以至不能正常传代。斑螟分索赤眼蜂的平均每雌寄生卵数为15.4粒,显著高于稻螟赤眼蜂的10.0粒与螟黄赤眼蜂的8.9粒。【结论】在室内人工培育的小菜蛾卵上繁殖的稻螟赤眼蜂与斑螟分索赤眼蜂对小菜蛾卵的寄生能力较强,尤其是斑螟分索赤眼蜂的内禀增长率r_m与小菜蛾的相近,适合以小菜蛾卵作为中间寄主扩大繁殖,稻螟赤眼蜂次之,而螟黄赤眼蜂并不适合以小菜蛾卵作为中间寄主。     

人工卵繁殖赤眼蜂实验种群生命表的研究

借助德国WILD公司生产的实体解剖镜(WILDN8),采用解剖法,统计赤眼蜂幼期存活率与成虫生殖力数据,编制了松毛虫赤眼蜂和拟澳洲赤眼蜂在人工卵上繁殖的实验种群生命表。结果表明,以人工卵作为寄主繁殖松毛虫赤眼蜂和拟澳洲赤眼蜂其实验种群对米蛾卵的寄生能力较强,内禀增长率rm分别为0．2442和0．3159,而松毛虫赤眼蜂比拟澳洲赤眼蜂更适于以人工卵作为寄主进行连代繁殖。松毛虫赤眼蜂在人工卵上单雌平均产卵量为61．5粒,卵、幼虫、蛹等幼期各期存活率分别为1．00、0．7020、0．9431;羽化率为0．7387。拟澳洲赤眼蜂在人工卵上繁殖1代。利用米蛾卵测其单雌平均产卵量为104．2粒,在人工卵内蜂卵、幼虫、蛹等幼期各期存活率分别为1．00、0．6454、0．9383;羽化率为0．7276。     

赤眼蜂的产卵行为

赤眼蜂能在多种宿主卵中产卵寄生,并且还具有在人工物体上产卵的习性。Holloway早在1912年即观察到秋葵植物汁滴干燥结膜后,广赤眼蜂(Trichogramma evanescen)企图在其表面产卵。Salt(1935)观察到赤眼蜂有企图将产卵器插入玻璃珠,水银滴,植物种子等表面的行为。Hagen等(1965)试制人工蜡卵饲喂草蛉幼虫时,偶然发现赤眼蜂可以产卵在这种蜡卵内。近年来,Rajendram和Hagen(1974)曾用上述蜡卵引诱赤眼蜂产卵于盐水或氨基酸溶液内。总之,赤眼蜂可在不少的非天然的宿主或物体上产卵,并且所报道产过卵的物体都是球形的。Salt(1937)认为决定赤眼蜂产卵的因素是化学感觉,夜蛾中促使赤眼蜂产卵的物质是22—25碳的直链烷烃成分。但影响产卵行为的各种因素尚缺乏详细的分析。     

不含昆虫物质的赤眼蜂幼虫人工饲料

为克服赤眼蜂繁蜂过程中天然奇主卵供应不足、成本过高的困难,我们进行了赤眼蜂幼虫人工饲料的研究。用鸡蛋黄、氨基酸溶液、鸡胚液、牛奶、蛋白陈等配制的人工饲料,以悬滴培养法培育松毛虫赤眼蜂 Trichogramma dendrolimi获得初步成功,化蛹率为20—40％,羽化率一般为5％、最高可达16％(1/3个体能展翅)。蜂体大小和发育速度与柞蚕卵培育的相似,但生活力略差,有交配能力并可在柞蚕卵内产卵,后代多为雌性。     

赤眼蜂人工模拟寄主卵的研究

本文报道松毛虫赤眼蜂Trichogramma dendrolimi在人工模拟寄主卵上产卵寄生并发育至成虫羽化出蜂的研究结果。对人工卵壳制作及人工培养液作了叙述,就赤眼蜂对人工培养液、人工卵壳的要求以及影响赤眼蜂在人工卵内发育的一些因素进行了讨论。     

赤眼蜂人工培育的研究

利用赤眼蜂防治农、林害虫,种类和防治面积上都在迅速增加,用柞蚕卵、蓖麻蚕卵繁育赤眼蜂,尚不能满足生产上迅速发展的需要。为此开展了赤眼蜂人工培育的研究。以鸡蛋盐液十30%核酪、鸡蚤盐液为营养液,人工培育赤眼蜂可由胚胎期发育至成蜂,其培育历期与用柞蚕卵培育的相仿。但由于人工饲料的营养不能满足需要,致使大部分个体在培育中停留在胚胎期或幼虫期。人工饲料本身的物理性状及培育届期的环境条件也很重要,因此,关于赤服蜂发育最好的营养和环境条件,仍有待以后继续研究改进。     

松毛虫赤眼蜂对假卵不同内含物的产卵反应

利用假卵测试了不同内含物对松毛虫赤眼蜂(Trichogramma dendrolimi)产卵的影响。结果表明在20种L型氨基酸中,只有亮氨酸、苯丙氨酸和异亮氨酸有促使产卵的效果,产人前二种氨基酸的卵量与产入柞蚕蛹血淋巴的卵量相似,后一种稍低于产入柞蚕蛹血淋巴的卵量。它们促使产卵的适宜浓度分别为200,400,320毫克/100毫升,较高浓度时,赤眼蜂依然能产入相当数量的卵。另外供试的17种氨基酸除在组氨酸内产入极少量卵以外,均未见在其它氨基酸内产卵。将亮氨酸、苯丙氨酸和组氨酸混合使用时,促使产卵效果更好,它们混合的最佳浓度分别为600,400,425毫克/100毫升,每假卵中的产卵量平均高达878.7粒,多于含血淋巴的假卵。 除氨基酸外,各种盐类(NaCl,KCl,MgSO₄等),糖类(葡萄糖、蔗糖、果糖、海藻糖等),混合有机酸,维生素均不促使产卵。在赤眼蜂人工培养中常用的成分如鸡胚提取液、牛奶、鸡蛋黄、牛血清、酵母液等促使产卵的效果很差,但与有促产卵效果的氨基酸混合时不降低后者的作用。基本培养液分别加入少量鸡胚提取液、牛奶或鸡蛋黄时,促使产卵的效果接近血淋巴。由于这样的培养液能使卵发育到幼虫,因而可用于大量蜂卵的收集。 假卵试验表明,赤眼蜂借产卵器上的感受器来识别假卵的内含物,触角对卵面探测所起的作用与产卵的关系很小。赤眼蜂可在大型假卵内产入大量卵,不受已产入卵的影响。但在小型假卵内的产卵是受影响的,这种差别可能是通过产卵器对卵内物质的感受而产生。     

诱导赤眼蜂寻找寄主卵的激素

寄生峰在田野寻找寄主卵要费很大力气,往往卵未产尽就死亡了,如果有一种物质能诱导寄生蜂很快找到奇主卵,可使寄生蜂发挥更大作用。 有人发现广赤眼蜂(Trichogramma evaneseens)在棉铃虫的卵粒上产卵,是因为棉铃虫蛾产卵时有鳞片掉落在卵上,发出气味引诱赤眼蜂上去产卵的。于是从夜蛾的鳞片中提出一种抽取物,经过分析研究,分离出四种不同化合物,都是直链碳氢化合物类,对赤眼蜂起引诱作用。根据此原理,进一步人工合成了四种引     

A STUDY ON THE OVIPOSITION SYNERGISTS FOR IN VITRO REARING TRICHOGRAMMA SPP.*

Abstract Solutions or suspensions of polyvinyl alcohol, gelatin, white latex, agar cassava powder, rice flour and starch were used as synergists to improve the oviposition of Trichogramma dendrolimi, T. confusum and T. evanescens with in vitro rearing method. The ovipositional response of Trichogramma spp. to these synergists was different. All the 7 preparations improved the trichogrammatid oviposition much on the artificial host “eggs”. Specific difference of trichogrammatid ovipositional response to these synergists was observed. So, the most effective and economical synergists can be used for increasing the efficiency of rearing different species of Trichogramma in vitro.     

Ovipositional Synergists and Artificial Diets for Trichogramma australicum Girault (Hymenoptera: Trichogrammatidae)

We studied ovipositional synergists and artificial diets for rearing Trichogramma australicum. Artificial "diet A" included 40% haemolymph of Helicoverpa armigera (Hübner) final instar larvae, 30% of a 10% malt solution in deionised water, 20% chicken egg yolk and 10% Neisenheimer's salt solution with 76 units Penicillin and 76 units Streptomycin/ml diet. Artificial "diet B" was identical except Grace's insect medium® replaced Neisenheimer's salt solution. the number of T. australicum larvae in artificial eggs filled with diet A and diet B was not significantly different (P > 0.05). Significantly fewer (P < 0.05) larvae developed to pupae and adults in artificial eggs filled with diet A. Quantity and quality of artificial diet affected the mortality of T. australicum larvae reared in vitro. Ovipositional synergists included 10% gelatine solution, 10% polyvinyl alcohol solution or 1% agar solution in deionised water. Synergist test-solutions were individually smeared on the external surface of artificial eggs (hemispherical depressions in plastic membrane). Eggs laid and number of T. australicum larvae produced were significantly higher in artificial eggs smeared with gelatine than artificial eggs smeared with polyvinyl alcohol, agar or non-smeared (control) eggs.     

Comparative Analysis of the Reproductive Attributes of Three Commercially-Produced Trichogramma Species (Hymenoptera: Trichogrammatidae)

The reproductive attributes of commercially-produced Trichogramma platneri, T. minutum and T. pretiosum reared from eggs of both Ephestia kuehniella and Sitotroga cerealella were monitored at 25 C. The age-specific fecundity, longevity and progeny sex ratio and adult size were determined for individual females from all six combinations of Trichogramma and rearing host, using E. kuehniella as the experimental host. Rearing host had a significant influence on the lifetime fecundity, 3-day fecundity and longevity of all three Trichogramma species. In general, the performance of T. minutum and T. pretiosum was better when reared from S. cerealella, but that of T. platneri was superior when reared from E. kuehniella. The lifetime fecundity of the Trichogramma species was linearly related to longevity and the ranking between species was T. pretiosum > T. minutum > T. platneri. The age-specific pattern of oviposition for T. platneri was distinctly precocious, with 40% of its lifetime fecundity oviposited on the first day, in contrast to 17-24% for the other two species. Progeny sex ratio over the lifetime of the Trichogramma females was slightly male biased and differed significantly from 0.5 for T. minutum and T. platneri. Daily sex ratio for parasitoids reared from the most productive rearing host was female biased only for the first day of oviposition for T. platneri in contrast to the first 5-6 days of oviposition for the other two species. There was little evidence that any of the reproductive attributes of these Trichogramma species, reared from small host eggs, was dependent on the size of the adult females.     

Effects of physiological condition and experience on oviposition behaviour of Trichogramma australicum Girault (Hymenoptera: Trichogrammatidae) on eggs of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae)

We produced an ethogram for and investigated effects of physiological condition and experience on oviposition behaviour for Trichogramma australicum on Helicoverpa armigera eggs. The data, coupled with the ethogram, have enabled the establishment of standard behavioural categories which can be used to assess acceptability of H. armigera eggs to T. australicum reared in vitro or in eggs of other species. Physiological and experiential conditions were investigated using females of three different ages (0–24, 24–48 and 48–72 h old), two types of sexual states (mated and unmated) and two types of ovipositional experience (naive and experienced). Durations of host finding, host examination, and post-ovipositional re-examination were reduced by oviposition experience and were shorter in younger females (0–48 h old). Oviposition experience reduces the duration of the drilling and percentage of females host-feeding. Being mated and young reduces the host examination time in subsequent oviposition bouts. Only ovipositionally inexperienced females host-feed before oviposition and host-fed more frequently than experienced individuals after oviposition. Host-feeding needs consideration for optimal culture.     

Oviposition Behaviour and Reproductive Performance of Trichogramma australicum Girault (Hymenoptera: Trichogrammatidae) Reared in Artificial Diet

Biotypes of Trichogramma australicum were reared on artificial diet "A", Helicoverpa armigera (Hübner) (natural host) eggs and Sitotroga cerealella (Olivier) (factitious host) eggs. We evaluated the quality of these biotypes by comparing their oviposition behaviour and reproductive performance on H. armigera eggs. Female T. australicum reared in vitro accepted H. armigera eggs for oviposition and displayed similar behavioural components of oviposition to females reared on natural hosts. However, handling time during oviposition in H. armigera eggs by females from in vitro rearing was significantly longer than handling time of females reared on the natural host because the females needed a significantly longer time for host feeding. In vitro reared females produced significantly more progeny and parasitised more H. armigera eggs than the females reared on S. cerealella. Improvements to the artificial diet for T. australicum may reduce handling time of the in vitro reared females and increase the female population. These improvements may include changes in nutrient content and size of the artificial eggs.     

Does the Polyphagous Egg Parasitoid <Emphasis Type="Italic">Trichogramma platneri</Emphasis> Nagarkatti (Hymenoptera: Trichogrammatidae) Display Behavioral Plasticity When Parasitizing Different Hosts?

Trichogramma platneri oviposition behavior on Amorbia cuneana egg masses was investigated under laboratory conditions. No relationship was detected between host surface area, number of edge turning on host eggs, and parasitoid clutch size. It was also observed that females parasitized the egg mass randomly by drilling into the egg mass and parasitizing individual eggs without using stereotypical behaviors to assess individual hosts (i.e. drumming and turning). However, clutch size did change due to ovipositional experience as naïve wasps with little or no ovipositional experience (<6 eggs parasitized), allocated significantly more progeny per host than wasps with longer ovipositional experience (24-h oviposition experience on a single egg mass). Moreover, naïve wasps parasitized significantly more eggs on the outer edge of the egg mass than experienced wasps. We contend that the physical characteristics of A. cuneana eggs and egg masses preclude T. platneri from completely discriminating between individual eggs. However, because T. platneri may be using kairomones from the egg mass, this described oviposition strategy remains effective.     

繁育寄主对三种赤眼蜂个体大小及抱卵量的影响

【目的】繁育寄主及成蜂日龄均会影响赤眼蜂体内的抱卵量,为了解繁育寄主及育出赤眼蜂个体大小与赤眼蜂抱卵量的关系,我们在室内研究了米蛾 Corcyra cephalonica（Stainton）卵和亚洲玉米螟 Ostrinia furnacalis （Güenée）卵作为繁育寄主对3种赤眼蜂雌蜂大小、抱卵量的影响。【方法】在光周期14L:10D、温度25±1℃的室内条件下,以米蛾卵、亚洲玉米螟卵作为繁育寄主,解剖不同日龄（羽化后12-72 h）松毛虫赤眼蜂 Trichogramma dendrolimi Matsumura、玉米螟赤眼蜂Trichogramma ostriniae Pang et Chen以及稻螟赤眼蜂Trichogramma japonicum Ashmead,观察雌蜂抱卵量,测量成蜂的大小,分析雌蜂抱卵量与其日龄、个体大小以及繁育寄主的关系【结果】在羽化后48 h以内,3种蜂的抱卵量随雌蜂日龄的增长而增加。羽化后72 h,除米蛾卵繁育的玉米螟赤眼蜂及玉米螟卵繁育的松毛虫赤眼蜂的抱卵量比羽化后48 h时略高外,其他组合均出现下降。以亚洲玉米螟卵为繁育寄主时的玉米螟赤眼蜂的抱卵量显著高于对应日龄的以米蛾卵为繁育寄主时的抱卵量,而松毛虫赤眼蜂（除羽化后12和24 h外）和稻螟赤眼蜂则相反;以米蛾卵为繁育寄主时,相同日龄的赤眼蜂中,松毛虫赤眼蜂抱卵量最高,稻螟赤眼蜂的抱卵量最低;以亚洲玉米螟卵为繁育寄主时,玉米螟赤眼蜂抱卵量最高,稻螟赤眼蜂抱卵量最低。雌蜂抱卵量与雌蜂个体大小呈线性正相关,相关程度随雌蜂日龄增加而更加明显。【结论】后足胫节长度可以作为评价松毛虫赤眼蜂、玉米螟赤眼蜂和稻螟赤眼蜂质量的指标。赤眼蜂抱卵量受繁育寄主、赤眼蜂蜂种及雌蜂日龄的显著影响,寄主的适合性也会影响育出赤眼蜂的大小,筛选合适的繁育寄主有助于提高赤眼蜂的质量。     

Experience acquisition by Trichogramma australicum Girault (Hymenoptera: Trichogrammatidae)

We investigated experience acquisition (alpha-conditioning) by females of the parasitoid Trichogramma australicum using host eggs of the noctuid moth Helicoverpa armigera (Hübner). We compared the acceptance of a host egg by females with different levels of ovipositional experience. The level of experience was designated using the standard oviposition sequence: (1) host contact (C); (2) host examination (E); (3) drilling (D); (4) full insertion (FI); and (5) oviposition (O). Each treatment consisted of a single experience level, but together these consituted a qualitative behavioural continuum of oviposition experience from naive (N) to experienced wasps. We found that host experience by adult T. australicum females can modify their behaviour. Mean duration of host finding was: N = C > E = D > FI = O. Mean duration of host examination and full insertion were: N = C > E = D = FI = O. Drilling was constant for all experience levels. Experience in drilling of the chorion during the previous host-exposure process represents a critical experience for a female and results in efficient handling and more ready acceptance of a subsequently encountered host egg.     

Host-foraging success of three species of Trichogramma (Hymenoptera: Trichogrammatidae) in a simulated retail environment

Three species of trichogrammatid egg parasitoids (Trichogramma deion Pinto & Oatman, Trichogramma ostriniae Pang & Chen, and Trichogramma pretiosum Riley) (Hymenoptera: Trichogrammatidae) were evaluated under laboratory conditions as potential biological control agents for the Indianmeal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), on retail shelves. A single shelving unit was used in each trial and a grid of sentinel egg disks was used to evaluate foraging success. The shelving consisted of pallet units with five shelves that were either bare or stocked with empty cereal boxes. In each replicate, approximately 500 female Trichogramma were released at the center of the shelving unit and allowed to forage for 48 h. Percentage of egg parasitism and percentage of host egg mortality were recorded after 7 d. Foraging success as well as the spatial pattern of parasitism differed significantly among the three Trichogramma species. Percentage of egg parasitism was approximately 4 times greater for T. deion than for T. ostriniae or T. pretiosum. The vertical distribution of parasitism by T. deion was also more uniform than for the other two species. In addition, the presence of packaging affected the foraging efficiency of T. ostriniae and T. pretiosum but not T. deion. Based on these findings, Trichogramma deion may be the best-suited candidate for augmentative biological control of P. interpunctella in retail stores, and a central release point of T. deion will likely provide adequate coverage of products on pallet-type shelving.     

Egg laying and development of Neotropical trichogrammatid species in artificial eggs

The goal of this work was to assess the suitability of two artificial diets for egg laying and development of Trichogramma atopovirilia Oatman & Platner, Trichogrammatoidea annulata De Santis, and Trichogramma bruni Nagaraja (all Hymenoptera: Trichogrammatidae). Additionally, the quality of wasps reared in vitro was compared with those reared in vivo. The ‘standard’ diet consisted of Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae) pupal holotissues (65%), chicken egg yolk (18%), fetal bovine serum (8.5%), lactalbumin hydrolysate (8.5%), and anticontaminants (0.3%). The ‘modified’ diet differed from the standard one only in the D. saccharalis pupal holotissues, that were replaced with Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae) ones. Females of the three trichogrammatid species laid eggs in artificial eggs containing any artificial medium, but the modified one received more eggs. Although the standard diet was accepted for oviposition by the three wasp species, no development occurred. On the modified diet, only T. atopovirilia was able to develop to adult emergence. Adult F1 were of a quality that was similar to insects reared in vivo.