小黑瓢虫生殖生物学、人工扩繁及人工饲料的研究
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摘要
小黑瓢虫成虫和幼虫均能捕食烟粉虱,是近年来国外引进的烟粉虱有效捕食性天敌。小黑瓢虫的规模化人工扩繁生产对充分发挥其控害作用,丰富我国天敌昆虫产品,提升烟粉虱生物防治水平,减少化学农药使用,保护农田生态环境,促进农产品质量安全具有重要的现实意义。本研究以小黑瓢虫的生殖生物学研究为基础,以适宜小黑瓢虫人工扩繁环境条件的探索为深入点,结合低温冷藏条件的研究和人工饲料的研制,完善较可行的小黑瓢虫人工大量扩繁的基础研究,为获得大量的小黑瓢虫应用于大田防治奠定基础。主要研究结论如下:
1.应用常规内部解剖方法,对小黑瓢虫生殖系统的结构和卵巢发育情况进行研究。结果表明:雌性生殖系统主要包括1对卵巢、1对侧输卵管、中输卵管、受精囊、交配囊、附腺、粘腺、外生殖片等;雄性生殖系统主要包括睾丸、1对储精管、1对输精管、射精管及1对附腺、弯管和阳基等;小黑瓢虫雌成虫的卵巢管数量恒定均为8根;根据外部形态上的特点以及卵黄沉积的情况,将小黑瓢虫的卵巢发育分为卵母分化期、卵母细胞生长期、卵黄形成及卵子成熟期3个时期,以及未分化期、生长发育初期、生长发育中期、生长发育后期、成熟产卵前期、成熟产卵期6个阶段。
2.小黑瓢虫交配活动的主导者是雄虫,雄虫性成熟后可进行交配,交配持续时间平均为18.57 min;雄虫的交配前期为4.97±0.07 d,雌虫的产卵前期为5.80±0.21 d;性成熟的成虫可全天交配,6:00～8:00 am为高峰时刻;雌虫交配后产卵,可全天产卵,白天的产卵量高于夜间的产卵量,毎雌平均可产卵379粒,产卵期77～111 d;交配次数的增加可以延长产卵期,提高小黑瓢虫的产卵量,但雌虫的寿命会下降;交配次数多的后代繁殖状况最佳。
3.营养水平、温度和湿度高低、光周期的长短等不同条件均会对小黑瓢虫的繁殖活动造成影响。小黑瓢虫适应性较强,虽然在低温(18℃)、高温(33℃)和低湿(45%RH)、高湿(95%RH)以及全光照(24L:0D)、全黑暗(0L:24D)等极端条件下也能进行交配,但繁殖效能较低。从食物、温度、湿度、光周期等环境因子对小黑瓢虫繁殖力的影响结果综合考虑,今后小黑瓢虫的饲养及人工扩繁应以烟粉虱卵为食料、温度24～27℃、湿度75～85%RH、光周期12L:12D～14L:10D为宜。
4.在饥饿条件下小黑瓢虫4龄幼虫和成虫耐饥力较强。幼虫的耐饥力随着虫龄的增加而增强,阶段死亡率高峰也随着虫龄的增加而推迟。小黑瓢虫的卵较适于短期(6 d以内)低温冷藏,10℃冷藏6 d孵化率为30%,卵在冷藏前让其在25℃下发育20 h后在进行冷藏能提高一定的孵化率,10 d后孵化率有32.22%;8℃是小黑瓢虫蛹短期内(10 d左右)冷藏的适宜温度,冷藏10 d后的羽化率为43.33%,随着冷藏天数的增加,羽化成虫的繁殖力呈下降趋势。
5.小黑瓢虫幼虫人工饲料的剂型以添加琼脂的凝胶状为优,其不易粘着幼虫,并有良好的成型效果,利于幼虫饲养。利用黄粉虫蛹为外源动物蛋白通过不同形式加工处理,配以酵母浸粉、蔗糖、蒸馏水等组分饲喂小黑瓢虫,2龄幼虫可以生长、化蛹、羽化完成世代。与取食烟粉虱的幼虫相比有以下几点差异:幼虫的整个发育历期明显延长,其中2龄幼虫发育历期相差不大,3、4龄幼虫和蛹期历期延长,而预蛹期则缩短;取食人工饲料各个龄期的存活率除了在预蛹期与取食烟粉虱的一致外,在幼虫2龄、3龄、4龄和蛹期这4个虫态的存活率均低于取食烟粉虱相应虫态的存活率;喂饲人工饲料后得到的3龄、4龄、预蛹和蛹各虫态的体长、体宽和头壳宽指数均较取食自然猎物的小;取食人工饲料后羽化得到小黑瓢虫成虫对烟粉虱卵的功能反应类型没有影响,均符合HollingⅡ模型,在攻击系数(a)和处理时间(Th),以及攻击率与处理时间比值(a /Th)和理论最大捕食量(1/Th)等指标上存在一定差异。从发育历期、各龄幼虫存活率、幼虫的体征、羽化率和成虫获得率和捕食功能反应综合结果表明,在黄粉虫蛹蛋白前处理中以脱脂粉的饲喂效果较佳。
Delphastus catalinae(Horn)is an important nature enemy of Bemisia tabaci imported from North America rencently. Both its adults and larvae can prey on different stages of whitefly. D.catalinae, an effective biological control agents of whitefly, has play an important role in the development of“Green Agriculture”in our country. A mass used D.catalinae in farmland is beneficial to reducing the use of chemical pesticide, protecting the ecological environment and improving the quality and safety of agricultural products. In this paper, the feasible basic research of mass rearing of D. catalinae was studied, including the reproductive biology of D.catalinae, suitable environmental conditions in artificial rearing, suitable stage to preservation in mass rearing, artificial diets of larvae. The main conclusions are as follows:
1. The reproductive system and ovarian development of D.catalinae was investigated by anatomical method. The results showed that the reproductive system of female adult was constituted by a pair of ovaries, a pair of oviducts, median oviduct, spermatheca, bursa copulatrix, accessory glands, sticky glands and genital plate. The reproductive system of male adult was constituted by a pair of testes, a pair of seminal vesicles, a pair of seminal ducts, ejaculatory duct, a pair of male accessory glands, shipo and penis. Ovarioles of D. catalinae are constantly 8 in adult developmental stages. Based on external morphology characteristics and the vitellines deposition conditions, the ovarian development in the adult stage of D.catalinae could be divided into 3 phases and 6 stages.
2. Mating was a male-dominated phenomenon. When sexually mature, the males initiated mating. A complete mating process took 18.57 min on average. A pre-mating period of 4.97±0.07d for the male and a pre-oviposting period of 5.80±0.21d for the female were observed. Multiple mating behavior occurred during the whole day and reached to a peak in the early moring at 6:00-8:00. After mating, female adults had many times oviposition behavior in one day, which laid more eggs during the day than at night. Each female adult laid average of 379 eggs on average and oviposition period lasted 77-111 days. With the increase of the number of mating, ovipsition period and fecundity increased but longevity of female decreased. Reproductive performance of D.catalinae was best after multiple matings.
3. The reproduction of D.catalinae was affected by nutrition, temperature, relative humidity(RH) and photoperiods. D.catalinae can mate and lay eggs in severe environment such as low-temperature(18℃), high-temperature(33℃), low-RH(45%), high-RH(95%) , 24 hours light and 24 hours dark, but the reproductive performance was worse. The suitable conditions of mass rearing for D.catalinae were 24-27℃, 75-85%RH, 12L:12D-14L:10D and the eggs of whitefly was the most favor food for D.catalinae.
4. There were great difference between the longevity of the larvae and adults of D.catalinae under starvation conditions. The longevity of the 4th instar larva and the adult was longer than the others. The starvation tolerance of the larva relied on their age. The suitable preservation conditions of eggs were described: earliest eggs firstly developing for 20h at 25℃, nextly storing for 10d at 10℃, lastly slowly developing to hatch at 25℃. Under this condition, hatching rate of eggs was 32.22% after 10d storing. The suitable preservation temperature of pupae was 8℃. The eclosion rate was 43.33% after 10d storing. Reproductive performance of D.catalinae was worse with the increase of storing days.
5. The gelatin type integrated with agar in the diet was the best form of the artificial diet for the larvae of D.catalinae. The type not only could hold water better but also was more molding that resistant the larvae adhere to diet. Yellow mealworm pupae was processed into four forms: freezing&drying powder, microwave powder, full-fat powder, defatted powder. Then made them into different artificial diets with other elements to rear the larvae of D.catalinae from the 2nd stage. D.catalinae fed with whitefly and reared with artificial diets were compared for the larval developmental duration, the larva survival rate, the indexes of body length, body width and head width and the predation functional response. The results indicated that fed with artificial diets the larval developmental duration became longer significantly, while there was no difference in the 2nd instar stage and shorter in prepupal stage. Except for the prepupal stage, that fed with artificial diets the larva survival rate of different stages was lower than those fed with natural prey. The 3rd, 4th instar larva, prepupae and pupae fed with artificial diets showed smaller body length, body width and head width than those fed with natural prey. The investigation showed that the predation functional responses of D.catalinae fed with artificial diets all corresponded with HollingⅡmodel, but the value of a/Th(the ratio of attack rate to handling time)in D.catalinae fed with whitefly was higher than those fed with artificial diets. By appraising on larva development time, larva survival rate, eclosion rate and adult obtaining rate, the defatted powder of yellow mealworm pupae would be the best form of the diet for the larva of D.catalinae.
1.应用常规内部解剖方法,对小黑瓢虫生殖系统的结构和卵巢发育情况进行研究。结果表明:雌性生殖系统主要包括1对卵巢、1对侧输卵管、中输卵管、受精囊、交配囊、附腺、粘腺、外生殖片等;雄性生殖系统主要包括睾丸、1对储精管、1对输精管、射精管及1对附腺、弯管和阳基等;小黑瓢虫雌成虫的卵巢管数量恒定均为8根;根据外部形态上的特点以及卵黄沉积的情况,将小黑瓢虫的卵巢发育分为卵母分化期、卵母细胞生长期、卵黄形成及卵子成熟期3个时期,以及未分化期、生长发育初期、生长发育中期、生长发育后期、成熟产卵前期、成熟产卵期6个阶段。
2.小黑瓢虫交配活动的主导者是雄虫,雄虫性成熟后可进行交配,交配持续时间平均为18.57 min;雄虫的交配前期为4.97±0.07 d,雌虫的产卵前期为5.80±0.21 d;性成熟的成虫可全天交配,6:00～8:00 am为高峰时刻;雌虫交配后产卵,可全天产卵,白天的产卵量高于夜间的产卵量,毎雌平均可产卵379粒,产卵期77～111 d;交配次数的增加可以延长产卵期,提高小黑瓢虫的产卵量,但雌虫的寿命会下降;交配次数多的后代繁殖状况最佳。
3.营养水平、温度和湿度高低、光周期的长短等不同条件均会对小黑瓢虫的繁殖活动造成影响。小黑瓢虫适应性较强,虽然在低温(18℃)、高温(33℃)和低湿(45%RH)、高湿(95%RH)以及全光照(24L:0D)、全黑暗(0L:24D)等极端条件下也能进行交配,但繁殖效能较低。从食物、温度、湿度、光周期等环境因子对小黑瓢虫繁殖力的影响结果综合考虑,今后小黑瓢虫的饲养及人工扩繁应以烟粉虱卵为食料、温度24～27℃、湿度75～85%RH、光周期12L:12D～14L:10D为宜。
4.在饥饿条件下小黑瓢虫4龄幼虫和成虫耐饥力较强。幼虫的耐饥力随着虫龄的增加而增强,阶段死亡率高峰也随着虫龄的增加而推迟。小黑瓢虫的卵较适于短期(6 d以内)低温冷藏,10℃冷藏6 d孵化率为30%,卵在冷藏前让其在25℃下发育20 h后在进行冷藏能提高一定的孵化率,10 d后孵化率有32.22%;8℃是小黑瓢虫蛹短期内(10 d左右)冷藏的适宜温度,冷藏10 d后的羽化率为43.33%,随着冷藏天数的增加,羽化成虫的繁殖力呈下降趋势。
5.小黑瓢虫幼虫人工饲料的剂型以添加琼脂的凝胶状为优,其不易粘着幼虫,并有良好的成型效果,利于幼虫饲养。利用黄粉虫蛹为外源动物蛋白通过不同形式加工处理,配以酵母浸粉、蔗糖、蒸馏水等组分饲喂小黑瓢虫,2龄幼虫可以生长、化蛹、羽化完成世代。与取食烟粉虱的幼虫相比有以下几点差异:幼虫的整个发育历期明显延长,其中2龄幼虫发育历期相差不大,3、4龄幼虫和蛹期历期延长,而预蛹期则缩短;取食人工饲料各个龄期的存活率除了在预蛹期与取食烟粉虱的一致外,在幼虫2龄、3龄、4龄和蛹期这4个虫态的存活率均低于取食烟粉虱相应虫态的存活率;喂饲人工饲料后得到的3龄、4龄、预蛹和蛹各虫态的体长、体宽和头壳宽指数均较取食自然猎物的小;取食人工饲料后羽化得到小黑瓢虫成虫对烟粉虱卵的功能反应类型没有影响,均符合HollingⅡ模型,在攻击系数(a)和处理时间(Th),以及攻击率与处理时间比值(a /Th)和理论最大捕食量(1/Th)等指标上存在一定差异。从发育历期、各龄幼虫存活率、幼虫的体征、羽化率和成虫获得率和捕食功能反应综合结果表明,在黄粉虫蛹蛋白前处理中以脱脂粉的饲喂效果较佳。
Delphastus catalinae(Horn)is an important nature enemy of Bemisia tabaci imported from North America rencently. Both its adults and larvae can prey on different stages of whitefly. D.catalinae, an effective biological control agents of whitefly, has play an important role in the development of“Green Agriculture”in our country. A mass used D.catalinae in farmland is beneficial to reducing the use of chemical pesticide, protecting the ecological environment and improving the quality and safety of agricultural products. In this paper, the feasible basic research of mass rearing of D. catalinae was studied, including the reproductive biology of D.catalinae, suitable environmental conditions in artificial rearing, suitable stage to preservation in mass rearing, artificial diets of larvae. The main conclusions are as follows:
1. The reproductive system and ovarian development of D.catalinae was investigated by anatomical method. The results showed that the reproductive system of female adult was constituted by a pair of ovaries, a pair of oviducts, median oviduct, spermatheca, bursa copulatrix, accessory glands, sticky glands and genital plate. The reproductive system of male adult was constituted by a pair of testes, a pair of seminal vesicles, a pair of seminal ducts, ejaculatory duct, a pair of male accessory glands, shipo and penis. Ovarioles of D. catalinae are constantly 8 in adult developmental stages. Based on external morphology characteristics and the vitellines deposition conditions, the ovarian development in the adult stage of D.catalinae could be divided into 3 phases and 6 stages.
2. Mating was a male-dominated phenomenon. When sexually mature, the males initiated mating. A complete mating process took 18.57 min on average. A pre-mating period of 4.97±0.07d for the male and a pre-oviposting period of 5.80±0.21d for the female were observed. Multiple mating behavior occurred during the whole day and reached to a peak in the early moring at 6:00-8:00. After mating, female adults had many times oviposition behavior in one day, which laid more eggs during the day than at night. Each female adult laid average of 379 eggs on average and oviposition period lasted 77-111 days. With the increase of the number of mating, ovipsition period and fecundity increased but longevity of female decreased. Reproductive performance of D.catalinae was best after multiple matings.
3. The reproduction of D.catalinae was affected by nutrition, temperature, relative humidity(RH) and photoperiods. D.catalinae can mate and lay eggs in severe environment such as low-temperature(18℃), high-temperature(33℃), low-RH(45%), high-RH(95%) , 24 hours light and 24 hours dark, but the reproductive performance was worse. The suitable conditions of mass rearing for D.catalinae were 24-27℃, 75-85%RH, 12L:12D-14L:10D and the eggs of whitefly was the most favor food for D.catalinae.
4. There were great difference between the longevity of the larvae and adults of D.catalinae under starvation conditions. The longevity of the 4th instar larva and the adult was longer than the others. The starvation tolerance of the larva relied on their age. The suitable preservation conditions of eggs were described: earliest eggs firstly developing for 20h at 25℃, nextly storing for 10d at 10℃, lastly slowly developing to hatch at 25℃. Under this condition, hatching rate of eggs was 32.22% after 10d storing. The suitable preservation temperature of pupae was 8℃. The eclosion rate was 43.33% after 10d storing. Reproductive performance of D.catalinae was worse with the increase of storing days.
5. The gelatin type integrated with agar in the diet was the best form of the artificial diet for the larvae of D.catalinae. The type not only could hold water better but also was more molding that resistant the larvae adhere to diet. Yellow mealworm pupae was processed into four forms: freezing&drying powder, microwave powder, full-fat powder, defatted powder. Then made them into different artificial diets with other elements to rear the larvae of D.catalinae from the 2nd stage. D.catalinae fed with whitefly and reared with artificial diets were compared for the larval developmental duration, the larva survival rate, the indexes of body length, body width and head width and the predation functional response. The results indicated that fed with artificial diets the larval developmental duration became longer significantly, while there was no difference in the 2nd instar stage and shorter in prepupal stage. Except for the prepupal stage, that fed with artificial diets the larva survival rate of different stages was lower than those fed with natural prey. The 3rd, 4th instar larva, prepupae and pupae fed with artificial diets showed smaller body length, body width and head width than those fed with natural prey. The investigation showed that the predation functional responses of D.catalinae fed with artificial diets all corresponded with HollingⅡmodel, but the value of a/Th(the ratio of attack rate to handling time)in D.catalinae fed with whitefly was higher than those fed with artificial diets. By appraising on larva development time, larva survival rate, eclosion rate and adult obtaining rate, the defatted powder of yellow mealworm pupae would be the best form of the diet for the larva of D.catalinae.
引文
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