【作者】 王福莲；

【导师】 田小海； 李传仁；

【作者基本信息】 长江大学 ， 作物保护学， 2018， 博士

【摘要】 柑橘大实蝇Bactrocera minax（Enderlein）为我国柑橘类重要害虫,主要以成虫期防治为主,包括食物诱杀和辐射不育技术。为了提高成虫诱杀防治的精准性和提高防治效果,为了解决柑橘大实蝇SIT技术应用的瓶颈问题,开展了柑橘大实蝇诱集适期、田内外分布、蛋白等食物对成虫适合性、蛹的低温保藏效应和辐射后成虫质量方面的研究。为准确简便预测柑橘大实蝇成虫在橘园内发生动态及其诱集适期,通过比对常温保蛹下的成虫羽化动态与橘园内的成虫诱集动态（以湖北宜都为例）,计算出橘园成虫诱集动态较常温保蛹下成虫羽化动态的相对滞后程度,并分析影响滞后程度的气候因素。结果表明柑橘大实蝇田间诱集盛期较羽化盛期具有相对稳定的滞后天数,始盛日、高峰日、盛末日的滞后天数分别为（15±1.35）d、（18.5±1.32）d、（22.5±1.55）d。各年度的滞后程度与滞后期间的温度、湿度、降雨量无显著相关性。本文为利用常温保蛹下的羽化预测橘园内柑橘大实蝇发生动态,预估橘园诱集适期提供范例,为提高柑橘大实蝇成虫诱杀防治的精准性提供依据。柑橘大实蝇在桔园内林木侧与非林木侧的分布特征因虫口密度的不同而不同。在低密度虫口下（2%和4%蛆果率）,桔园内林木侧虫量分布比例显著高于非林木侧虫量比例。在较高虫口密度下（蛆果率20%）,桔园内林木侧与非林木侧诱集点的虫量分布比例无显著差异。不同虫量,雌虫和雄虫在林木侧和非林木侧的分布情况与总虫一致,雌雄虫无显著差异。从柑橘大实蝇在桔园内外的发生动态来看,柑橘大实蝇在桔园内发生一个高峰,高峰期虫口密度高于桔园外林木带。在桔园外林木带发生两个高峰,第一高峰与桔园内高峰吻合,第二高峰在桔园内高峰结束后。在柑橘大实蝇的起始诱集期和诱集末期,以外侧林木边带虫口密度较大。发生前期主要分布在柑橘外20 m处,在发生高峰期,主要分布在桔园内50m和内10m处,在发生末期主要分布在桔园外10m处。在柑橘大实蝇发生起始期,主要分布在外侧的林木边带,经过一次向内的扩散后分布于园内外。发生高峰期维持园内外分布状态。其后经过一次向外和一次向内扩散后,于发生末期逐步向外扩散。柑橘大实蝇向内或向外扩散,大多为雌虫先行扩散,雄虫随后。结果可为提高诱杀布点的合理性提供依据。柑橘大实蝇其卵、幼虫和蛹较为隐蔽,多采用成虫诱杀。成虫诱杀基于柑橘大实蝇成虫在性成熟期需要大量补充营养,对糖和蛋白表现出强烈趋性。目前,柑橘大实蝇成虫诱杀多在羽化后18-23d进行,而关于初羽化成虫取食选择性的研究较少。我们对初羽化成虫（1-7日龄）在水和蜂蜜之间、蔗糖中添加蛋白前后、在不同生境中的蜂蜜之间以及保鲜剂在蜂蜜中添加前后等的取食选择性进行研究。结果表明,初羽化的柑橘大实蝇雌虫在2-3d、雄虫在3-4d,表现出对水的较高选择性。蛋白添加后比添加前的选择性显著降低。在不同生境的蜂蜜之间,雄虫对乔木和灌木生境的五倍子蜂蜜和枣树蜂蜜表现出显著高的选择性。另外,雌虫对添加保鲜剂（山梨酸和尼泊金）的蜂蜜的的取食选择性显著降低,雄虫对添加尼泊金的蜂蜜取食选择性显著降低。本研究明确柑橘大实蝇初羽化成虫对糖和水、糖和蛋白、添加防腐剂的蜂蜜和不同生境源蜂蜜的取食选择性,利于柑橘大实蝇初羽化成虫的诱剂开发和增强诱剂的防治效果,为提前成虫诱集期、提高诱集效果、降低诱剂使用成本和合理设置诱集点等提供依据。柑橘大实蝇人工饲养技术不尽完善。为了明确添加蛋白对柑橘大实蝇的影响,研究了取食四种食物F0（水）、F1（蔗糖）、F2（蔗糖+酵母）和F3（蔗糖+蛋白胨）时柑橘大实蝇的生理适合性。结果表明,取食F2和F3时,柑橘大实蝇的寿命更长,分别为40.1d和32.8d;死亡率较低,死亡高峰后延（分别为5.6d和4.3d）;交配次数增多（单雌一生交配次数分别为8.1次和5.3次,单雄一生交配次数分别为4.7次和7.3次）,交配期更长（分别持续42d和34d）。此外,雌虫卵巢发育更快,怀卵更多（单雌94.8粒和77.3粒）,产卵量更大（单雌产卵量分别为63.2粒和19.3粒）。单虫饲喂蔗糖时,柑橘大实蝇虽然怀卵,但没有产卵,而且体内总糖含量和海藻糖含量较高。结果可为柑橘大实蝇人工饲养和SIT控制技术的研究提供基础和虫源。温度是影响柑橘大实蝇越冬蛹羽化的关键因子。蛹期保藏是拓展柑橘大实蝇虫源和保证虫源持续性的重要途径。而了解低温对柑橘大实蝇的影响是柑橘大实蝇虫源保藏的基础。1)研究了柑橘大实蝇0级蛹在10℃下冷藏不同天数后,对1-4级各级蛹发育高峰和成虫羽化高峰的延迟效应,结果表明:采用10℃对柑橘大实蝇0级蛹进行冷藏,以40日以内为宜,冷藏35日时成虫的羽化率与对照无差异,此时其1级、2级、3级、4级蛹发育高峰延迟15d、18d、24d、15d,成虫羽化高峰期可以推后11d。2)研究了柑橘大实蝇1-4级蛹在短时温和低温变化温度体系（25℃-25℃-25℃,25℃-15℃-25℃,25℃-20℃-25℃）下的存活情况,结果表明,柑橘大实蝇1级蛹在经25℃-20℃-25℃,25℃-15℃-25℃两个短时温和低温体系下处理2,4和8h,均促进其羽化。2-3级蛹在25℃-15℃-25℃,25℃-10℃-25℃的短时温和低温体系下处理4h和8h,促进其羽化。4级蛹在25℃-20℃-25℃,25℃-15℃-25℃,25℃-10℃-25℃三个短时温和低温体系下处理2,4,8h均有促进其羽化。可见,特定短时温和低温体系处理柑橘大实蝇各级蛹,利于其存活。3)为了解短时低温对柑橘大实蝇1-4级蛹恒高温代价的弥补效应,以25℃恒高温为基础,比较了相对低温10、15、20℃短时（2、4、8h）变温处理柑橘大实蝇1-4级蛹后其成虫羽化增长幅度。相对于恒温25℃,经15℃和20℃短时（2⁸h）变温处理1级蛹后成虫羽化增长率为14%⁴7%,经10℃和15℃短时（2、4、8h）变温和20℃短时（4h）变温处理2-3级蛹后成虫羽化增长率为2%⁴5%,经10、15℃和20℃短时（2、4、8h）变温处理4级蛹后成虫羽化增长率为28%⁴6%,均表现为正增长,产生了恒温代价弥补正效应。4级蛹经10、15、20℃变温处理2h和8h,对其恒温代价弥补正效应均较2-3级蛹的大。总之,特定时长低温对特定蛹级产生恒温弥补正效应,不同蛹级间效应有差异。本研究结果为室内柑橘大实蝇保存,及春季快速回暖或田间倒春寒情况下柑橘大实蝇种群动态的预测预报提供依据。为解决制约柑橘大实蝇SIT技术应用的虫源问题,提高辐射虫质量。开展了柑橘大实蝇辐照蛹级的扩展和较高不育剂量⁶⁰Coγ射线辐照对柑橘大实蝇存活的影响研究,为提高柑橘大实蝇辐照虫的质量提供依据。结果表明:1)1级蛹、2级蛹和3级蛹辐射后（90Gy）的羽化率与自然虫或4级蛹辐射后的羽化率均无显著差异。1-4级蛹平均雌雄性比与自然虫无差异,羽化高峰与自然虫和4级辐照蛹的成虫羽化高峰基本一致。除3级蛹辐照雌虫外,其他辐照成虫与自然成虫的交配次数和交配持续时长无差异。综合田间实际应用情况,考虑把柑橘大实蝇辐照蛹期扩展为2-3级蛹高峰期,此时为1-4级混合蛹。2)采用较高不育剂量（100Gy、110Gy）⁶⁰Coγ射线辐照后,柑橘大实蝇雌雄虫的死亡率与自然虫无差异。雌雄虫的死亡高峰与自然虫比较发生了提前和延后现象,但辐照虫和自然虫的死亡高峰均发生在10日龄前。雄虫交配前期的校正死亡率分别为24.53%和39.62%,均显著高于雌虫。采用100Gy和110Gy较高不育剂量的⁶⁰Coγ射线辐照柑橘大实蝇1-4级混合蛹后,对柑橘大实蝇成虫的存活影响不大。更多还原

【Abstract】 Bactrocera minax（Enderlein）,a major citrus pest in China,is mainly controlled by trapping and SIT in adult stage.The optimum trapping period,the distribution inside and outside of the orchard,the physiological function of proteins on adult,the cool temperature starage of pupae,and the quality of erradiated adults of the fly were study to improve the trapped accuracy and effect in field,to solve the apply of SIT limitted by lack of insects.In order to monitor the adult dynamics and predict its optimal trapping period in the orchards,we compared the dynamics of adult eclosion and field trapping in Yidu,and calculated the interval of the peak period between eclosion and capture.Meanwhile,the environmental factors during the interval,including the temperature,humidity,and rainfall were analyzed.The results showed that the interval was relatively stable between eclosion and trapping.The interval of the beginning,apex and ending of the adult peak period between eclosion and trapping were 15±1.35 days,18.5±1.32days and 22.5±1.55 days,respectively.There was no significant correlation between the interval days and the temperature,humidity,and rainfall during the interval.In the way to monitor dynamics of eclosion at normal temperature,and then foreseeing the optimum trapping period of B.minax in orchard,this study can provide the basis for enhancing the accuracy of trapping.The distribution characteristics of Chinese citrus fruit flies,B minax,in the orchard on the side of wood were different from the non-forest side according to the population density.When the density of adults population were low（rate of maggot fruit at 2%-4%）,adults was much more in the wood side than that in the non-wood side.But,when at higher densities（rate of maggot fruit at 20%）,there was no significant difference with adults in both sides.There was a coincidence with the females,males and total adults distributed in both sides,and there was no significant difference with females and males.From the occurrence dynamics of adults inside and outside the orange orchard,the citrus fruit fly has a peak in the orange orchard in which the adult density was higher in the internal orchard than that of external orchard.There are two peaks occurring to external orchard that the first peak coincided with the peak of internal orchard and the second peak is after the internal.During the start and end of trapping period of adults,the population density of external orchard was greater than that of external.The pre-infesting period was mainly distributed at 20 m outside the orchard.The peak period was mainly distributed at 50 m and 10 m inside the orchard and the end period was mainly distributed at 10 m outside the orchard.In the initial field occurrence,adults was mainly distributed in the adjacent wood and then after once inward diffusion,then distributed inside and outside of the orange orchard.The occurrence of peak period maintained the similar distribution in the internal and external orchard.Finally,after one outward and one inward diffusion,adults were gradually move towards the external orchard at the end of occurrence period.Whether the inward or outward diffusion of adults,it was mostly that females went firstly and males followed.The eggs,larvae and pupae of Chinese citrus fruit fly are very hard to find due to their shielding in fruit or in soil and therefore,baits spray for adults is the main management method in the field（baits trapping）。Baits spray is based on characteristic behavior of adult flies that displays a strong propensity to sugar and protein.However,the main weakness with this method is that the trapping period is commonly against the mature adults and little knowledges are known about the feeding selectivity of the early age of adult flies,such as newly-emerged infants.In this study,we investigated the feeding selectivity of newly-emerged B.minax for diets,by observing the number of infantile fruit flies to imbibe on water or honey,honey with or without protein,honey with or without antiseptics（sorbic acid or nipagin）,and honeys from different habitats,respectively.The results showed that there were no significant effects on the feeding selectivity between water and honey,although females at the 2nd and 3rd day and males at the 4th day after eclosion had displayed clearly feeding preferences to the water than honey.The feeding selectivity of honey after adding protein became visibly weakened for newly-emerged females not males.And the feeding selectivity of female infants to honey was greatly reduced with adding nipagin and sorbic acid,and that of male infants to honey significantly reduced only with adding nipagin in honey.In addition,for males,they fed gallnut honey from tree and shrub habitat more selectively than both motherwort honey and rape honey from herb habitat and honeysuckle honey from vine habitat,and more jujube honey from tree and shrub habitat than motherwort honey from herb habitat,but,there were no significant effects on feeding selectivity to these honeys from diverse habitats for females.The study provided us new thoughts for pest management of B.minax by the way of aiming at the newly-emerged adults,keeping baits effective and active,and set proper trapping station.Artificial rearing technology of B.minax adult is not well documented to date.In this study,we tried to determine if supplementing proteins to the adult diet could result in the enhancement of some fitness parameters of B.minax.Four feeds with varying protein source were provided as F0（water）,F1（sucrose）,F2（sucrose+yeast）,F3（sucrose+peptone）.F0 and F1 being the control,F2 and F3 were protein food types.The results showed that adults fed by F2 and F3 lived longer with 40.1 days and 32.8 days,respectively,had reduced death rates（death peaks were delayed for 5.6 days and 4.3 days,respectively）,increased mating frequencies（8.1 and 5.3 per females,4.7and 7.3 per males,respectively）,and longer mating durations（with 42days and 34 days）.In addition,females recorded an increased adult ovary development,more egg load（with 94.8 and 77.3 brood eggs per ovary）and to greater oviposition rates of 63.2 eggs/female and 19.3 eggs/female.Feed with F1,although there were eggs developed in ovary,but no eggs laid outside,and content of carbohydrates and threhalose were higher inside adult of B.minax.Based on our results,protein supplements enhanced B.minax survival,mating and fecundity.This study does not only provide basic knowledge to implement artificial rearing of B.minax,but also deepens our understanding on its physiology that could be used to enhance the management of the pest.Temperature is the main factor on eclosion of B minax pupae.Pupae preserve is one main manner to gain enough B.minax for SIT or studing on control continuous.How low temperature effect on the fruit fly must be clear when pupae preserve were done.1)Development delay effects of 1-4grade pupae with 0-grade pupae storage at 10℃for different duration were studied.The results showed that:storage with 0-grade pupae at 10℃,when the cold duration shorter than 40d,the eclosions had no significant difference with control.If cold storage duration exactly be 35d,the delay duration of pupae development peak in 1-grade,2-grade,3-grade and 4-grade were 15d,18d,24d and 15d.the delay duration of eclosion peak were11d.2)Survival level were evaluate by deal 1-4 grade pupae with25℃-25℃-25℃,25℃-20℃-25℃,25℃-15℃-25℃,25℃-10℃-25℃low temperature regime in short-time.Results showed that:1-grade pupae exposed to 25℃-20℃-25℃,25℃-15℃-25℃for 2h,4h and 8h could lead to more eclosion.The same as 2-3 grade pupae exposed to 25℃-15℃-25℃,25℃-10℃-25℃for 4h or 8h,4-grade pupae exposed to 25℃-20℃-25℃,25℃-15℃-25℃,25℃-10℃-25℃for 2h,4h or 8h.when 1,2,3 or 4 grade pupae exopsed to special short time regime will benefit to the survival.3)To demonstrate the trade-off effects on constant temperature cost of the 1-4st stages pupae of B.minax,the 1-4st stages pupae were exposed to series lower temperature（10℃,15℃,20℃,based on constant temperature of25℃）for 2h,4h and 8h and the adult eclosion growth were calculated.The results showed that:The eclosion growth rate,were 14%-47%when the 1st stage pupae were exposed to 15℃or 20℃for 2h,4h and 8h,were 2%-45%when the 2-3st stages pupae were exposed to 10 or 15℃for 2h,4h and 8h and 20℃for 4h,were 28%-46%when the 4th stage pupae were exposed to10℃,15℃or 20℃for 2h,4h,8h,with the positive growth.It indicated that there were recover effects on the constant temperature cost.The recover effects of the 4th stage pupae exposed to 10℃,15℃and 20℃for2h and 8h was higher significantly than these of the 2-3st stages pupae.It was concluded that certain stage pupae exposed to exact lower temperature with particular exposure time can produce recover effects on constant temperature cost and there be significantly recover difference between different stage pupae.The results could be referenced for the B.minax pupae preserve in laboratory and adult population forecasting in the field with rapid warm in early spring or cold spell in later spring.Aim to break the adult amount and quality restriction of SIT applied in field,the suitability of irradiated pupae stage and effect of higher sterility dose on B.minax were studied.Results showed that:1)Irradiated by⁶⁰Coγray at 1-grade,2-grade and 3-grade papae stage,the eclosion rate,eclosion peak have no difference singnicantly with the index of 4-grade or no irradiate natural flies.The mating frequence and mating duration have no singnicant difference except females from 3-grade irradiated pupae.So,for the apply in field,the suitable stage of irradiated could be at the peak of 2-3 grade pupae（mixed with 1-4 grade pupae）.2)irradiated by100Gy、110Gy⁶⁰Coγray on 1-4 grade pupae,mortality rates of male and female have no difference with natural male’s and female’s.although there are advance or delay of death peak,but no death peak occurred exceed 10 ages.Correct mortality rates（24.53%,39.62%）of irradiated males were higher significantly than females.In totally,the survival of adults from 1-4grade pupae irradiated by 100Gy、110Gy⁶⁰Coγray were no influenced.更多还原