应用寄生蜂和不育雄虫防控田间橘小实蝇

食品安全已成为当今社会广受关注的问题。尽管国内的一些学者就阿里山潜蝇茧蜂[Fopius arisanus(Sonan)]和橘小实蝇SIT技术分别开展了不同程度的研究,然而关于应用该蜂和不育雄虫对橘小实蝇[Bactrocera dorsalis(Hendel)]进行田间联合防治的研究还未见相关报道。为此,为探讨天敌昆虫和雄性不育技术对橘小实蝇的田间综合治理(IPM)技术,开展了阿里山潜蝇茧蜂和橘小实蝇不育雄虫的田间释放试验:首先应用性引诱剂降低田间野生雄虫数量;而后,释放橘小实蝇不育雄虫,进一步降低田间橘小实蝇种群数量;最后,释放阿里山潜蝇茧蜂对田间残余的橘小实蝇卵进行追踪寄生,以达到持续控制的目的。通过监测整个防控期田间橘小实蝇的种群数量以及果实受害率和防治效果作为评价指标评价该防治方法的综合防治效果。结果表明:应用阿里山潜蝇茧蜂、橘小实蝇不育雄虫并配合引诱剂的综合措施对橘小实蝇的种群数量有明显控制作用,田间总体防治效果可达90%左右。此外,提出完善橘小实蝇田间防控措施的建议,并提出该研究的不足以及有待进一步开展研究的方向。     

Purification and biochemical characterization of glutathione S-transferases from four field populations of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

Glutathione S-transferases (GSTs) are a group of detoxification enzymes that catalyze the nucleophilic addition of glutathione to a wide variety of endogenous and xenobiotic compounds. In this study, GSTs were purified from four field populations of Bactrocera dorsalis with different insecticide susceptibilities by glutathione-agarose affinity chromatography. The populations were collected from Dongguan (DG) and Guangzhou (GZ) of the Guangdong Province, Haikou of the Hainan province (HN), and Kunming of the Yunnan province (YN), China. Differences in GST characteristics among the four populations were studied using purified enzyme samples through comparative SDS-PAGE, kinetic, and inhibition experiments. The specific activities of the purified enzymes were similar, but the purification yield of the GZ population (31.54%) was the lowest. SDS-PAGE analysis showed only one band at approximately 23 kDa for these four populations. Kinetic analyses showed that the affinities of the purified GSTs from the GZ and YN populations for 1-chloro-2.4-dinitrobenzene (CDNB) were much higher than those of GSTs from the other two populations, whereas the HN population had the highest catalytic capability in terms of V(max) value. The optimum temperature for CDNB conjugation was 37 °C and the optimum pH was 7.5 in all four populations. Inhibition kinetics showed that ethacrynic acid, diethyl maleate, tetraethylthiuram disulfide, curcumin, bromosulfalein, and β-cypermethrin had excellent inhibitory effects on GSTs in the four populations of B. dorsalis, but the low inhibitory effects of malathion and avermectin did not differ between populations. These results suggest that GSTs may have a role in detoxification of β-cypermethrin in B. dorsalis.     

Dietary wheat germ oil influences gene expression in larvae and eggs of the oriental fruit fly

Changes in animal nutrition, particularly essential dietary components, alter global gene expression patterns. Our goal is to identify molecular markers that serve as early indicators of the quality of insect culture media. Markers of deficient culture media will increase the efficiency of developing optimal systems for mass rearing beneficial insects and some pest species because decisions on culture media quality can be made without waiting through one or several life cycles. The objective of our current study is to discover molecular markers of essential dietary lipid deficiency in the oriental fruit fly, Bactrocera dorsalis. We reared groups of fruit flies separately on media either devoid of or supplemented with wheat germ oil (WGO) and analyzed gene expression in third instar larvae and F(1) eggs using 2D electrophoresis. Gel densitometry revealed significant changes in expression levels of genes encoding eight proteins in larvae and 22 proteins in eggs. We identified these proteins by using mass spectrometry (MALDI TOF/TOF) and bioinformatic analyses of the protein sequences. Among these, we identified one gene encoding the receptor of activated C Kinase 1 (RACK1) that increased in expression by 6.8-fold in eggs from adults that were reared as larvae on media supplemented with WGO. RACK1 is an essential component of at least three intracellular signal transduction pathways, making it a good molecular marker candidate of lipid deficiency in fruit flies and possibly many other insect species.     

CDNA cloning and characterization of S6 kinase and its effect on yolk protein gene expression in the oriental fruit fly Bactrocera dorsalis (Hendel)

p70 S6 kinase (S6K), a serine/threonine protein kinase, is a downstream target of target of rapamycin (TOR) gene and an important regulator of protein synthesis responsible for cell growth and reproduction. In this study, a S6K gene, named BdS6K (GenBank Accession No. GQ203802), was isolated from the oriental fruit fly Bactrocera dorsalis (Hendel). Quantitative RT-PCR showed that BdS6K mRNA is expressed at a higher level in egg than in other developmental stages, as well as in ovary than in fat body. Downregulation of BdS6K activity by rapamycin treatment in larval stage resulted in the developmental defects of larvae, pupae, and adults, with a reduced yolk protein (YP) expression in the fat body throughout the first reproductive cycle with a substantial reduction in ovary size, and also repressed the egg development in female fruit fly. Knockdown of BdS6K gene by RNA interference in the adult significantly decreased the YP expression. These observations support the involvement of BdS6K signaling in the regulation of the YP synthesis and egg development in B. dorsalis.     

蝇蛹金小蜂对桔小实蝇蛹的寄生功能反应

在温度25±1℃、相对湿度70±5%、光照周期（L∶D）14 h∶10 h条件下,研究了蝇蛹金小蜂对桔小实蝇蛹的寄生功能反应及干扰效应。结果表明,寄主密度、寄主日龄均影响寄生蜂的寄生效能。蝇蛹金小蜂对桔小实蝇2日龄（N=2）、4日龄（N=4）蛹的寄生功能反应均符合HollingⅡ模型,其方程分别为Na= 0.4494N2/ (1+ 0.0294N2)、Na= 0.5586N4/ (1+ 0.0253N4)。24h内单头雌成蜂最多可寄生2日龄、4日龄蝇蛹数量分别为15.29、22.08头。自身密度对蝇蛹金小蜂寄生产生一定的干扰效应,其干扰效应符合Hassell－Varley模型（a= 0.0719P－0.2526）,表明蝇蛹金小蜂雌蜂的发现域随着自身密度的增加而逐渐变小,雌蜂个体间干扰效应降低了寄生效能。     

Phylogeographic patterns and cryptic speciation across oceanographic barriers in South African intertidal fishes

Biogeographic boundaries are the meeting zone of broadly distributed faunas, or the actual cause of a faunal break. In the latter case, closely related sister species should be found across such a boundary. To achieve such a situation, preliminary stages are expected, where phylogeographic breaks followed by genetic cryptic speciation would be observed. Biogeographic boundaries, in the Cape Point/Cape Agulhas region of southern Africa, offer an ideal system to test such predictions. Here, we studied two intertidal clinid fish species that are endemic to southern Africa, Clinus superciliosus (n = 127) and Muraenoclinus dorsalis (n = 114). Using mitochondrial control region, 16S rRNA, 12S rRNA and NADH2 genes and the nuclear rhodopsin and the first intron of the S7 ribosomal protein gene, we show both phylogeographic breaks and likely cryptic speciation in each species. Pairwise Φ(st) results suggest population genetic structuring for both species, with higher levels for M. dorsalis (Φ(st) = 0.34-0.93) than for C. superciliosus (Φ(st) = 0.1-0.74). Further, we recover two and three distinct lineages within M. dorsalis and C. superciliosus, respectively. Phylogenetic topologies, concordance between nuclear and mitochondrial markers and levels of sequence divergence, which are consistent with closely related sister species pairs, suggest the presence of cryptic species. Our results therefore meet the expectation for reduced gene flow at a biogeographic barrier, which translates into significant genetic breaks and cryptic sister species.     

拒避-诱杀组合技术对橘小实蝇产卵的影响

本文研究了以矿物油(mineral spray oil)乳剂为保护剂、"猎蝇"蛋白质毒饵为诱杀剂组成的拒避-诱杀组合技术对橘小实蝇Bactrocera dorsalis (Hendel)产卵的控制作用。结果显示:橘小实蝇在采用拒避-诱杀组合技术保护的供试香蕉上平均产卵孔数为0.000±0.000个,果实完好率达到100%,而单独使用"猎蝇"蛋白质毒饵的平均产卵孔数为27.000±1.732个,果实完好率仅为5.55%,表明该组合技术对橘小实蝇产卵的控制效果显著。在100倍～300倍浓度范围内,矿物油浓度越高,拒避-诱杀组合对橘小实蝇产卵的控制效果越好;随着虫口密度的增加,毒饵使用量和施用面积也须相应增加,才能取得更好的防治效果。     

气候因子对橘小实蝇生长发育及地理分布的影响

文章概述橘小实蝇Bactrocera dorsalis(Hendel)生活史、地理分布和生态要求的基础上,分析温度、湿度、降雨以及气候因子与橘小实蝇生长发育的关系,以及对橘小实蝇地理分布的影响。综述结果表明:喜湿热,厌干冷是橘小实蝇主要的生态学特征,低温与干旱不利于其生长发育;温度是橘小实蝇纬度分布中最主要的影响因子,冬季低温是中高纬度分布的主要限制因子,夏季高温是低纬度地区的限制因素之一,降雨影响橘小实蝇在经度上的分布范围。     

植物果实、颜色和形状对橘小实蝇产卵选择的影响

橘小实蝇Bactrocera dorsalis(Hendel)是近几年入侵江苏南部地区,严重危害当地水果、蔬菜的重要害虫。为深入了解其在该地区果树、蔬菜上的产卵特性,进行植物果实、颜色和形状对其产卵选择影响的研究。实验室中"纸杯法"的产卵选择试验结果表明,橘小实蝇对不同水果的产卵偏好性由高到低依次为:柑橘、香蕉、芒果、枣、石榴、苹果、桃子、梨、李、葡萄、柿;对不同蔬菜的产卵偏好性由高到低依次为:丝瓜、豇豆、苦瓜、黄瓜、茄子、南瓜、青椒、番茄。对不同寄主颜色和形状的产卵选择特点为,雌虫常选择桔黄色、绿色,以及圆球形的寄主产卵,而较少选择黑色、方形的寄主产卵。从试验结果可以看出,寄主果实散发出的气味,及其颜色和形状对橘小实蝇的产卵选择有影响。     

橘小实蝇交配行为观察

在实验室内对橘小实蝇Bactrocera dorsalis（Hendel）的交配行为进行了观察。描述了橘小实蝇交配中雄虫的“求偶场”,雄雄相遇、雌雌相遇、雌雄相遇时的相互反应,雌雄虫的交配行为与交配后行为。