不同寄主植物对桃小食心虫生长发育和繁殖的影响

为探索寄主植物对桃小食心虫Carposina sasakii生长发育和繁殖的影响, 在室内温度23±1℃, 相对湿度80%±7%, 光周期15L∶9D条件下, 测定了桃小食心虫在杏Armeniaca vulgaris、 李Prunus salicina、 桃Amygdalus persica、 枣Ziziphus jujuba、 苹果Malus pumila和梨Pyrus sorotina上各发育阶段的历期、 存活率和/或产卵量, 并组建了桃小食心虫在各寄主植物上的生命表。结果表明： 桃小食心虫的生长发育和繁殖在不同寄主植物间存在显著差异。幼虫的发育历期以李为最短（12.48 d）, 梨为最长（19.15 d）; 整个幼虫期的存活率以李为最高（50.54%）, 梨为最低（17.91%）; 单雌平均产卵量以枣（214.50粒/雌）和桃（197.94粒/雌）最高。生命表分析结果表明, 净生殖率R₀以枣（117.49）为最大, 平均世代周期T则以梨（41.31 d）和苹果（41.51 d）最长, 内禀增长率r_m以李（0.1294）为最高, 其次为枣（0.1201）和杏（0.1128）。这些结果有助于深入了解该虫在不同寄主植物上的种群动态。     

Sexual dimorphism of adult labial palps of the peach fruit moth Carposina sasakii Matsumura (Lepidoptera: Carposinidae) with notes on their sensilla

The labial palps and their sensilla of the peach fruit moth Carposina sasakii Matsumura, a serious pest of pome fruits in eastern Asia, were investigated using light microscopy and scanning and transmission electron microscopy. The labial palps are three‐segmented and exhibit distinct sexual dimorphism in length: much longer in the female than in the male. Four types of sensilla were found on the labial palps: campaniform, squamiform, chaetic and flattened sensilla. The campaniform sensilla are present on the first segment alone. The squamiform sensilla are located on all the three segments. The chaetic sensilla are mainly present on the third segment. The flattened sensilla are grooved with wall pores and situated in a labial palp‐pit organ, which is located at the apex of the distal segment of the labial palp. The sexual dimorphism of the labial palp and the labial palp‐pit organ was briefly discussed.     

桃蛀果蛾寄主生物型分化的RAPD分析

以山茱萸蛀果蛾Carposina coreana Kim为外群,用44个随机引物对苹果、山楂、桃、杏、枣和酸枣等6种寄主植物上的桃蛀果蛾Carposina sasakii Matsumura进行了RAPD 分析。结果表明,不同寄主上的桃蛀果蛾之间遗传差异出现了较大的分化,桃蛀果蛾6个寄主生物型之间的平均遗传距离在0.040～0.637之间,尤其是杏生物型分化最大。经聚类分析,可将不同寄主上的桃蛀果蛾明显分成两个类群,杏生物型为一群,其余为一群。两个类群间遗传距离为0.465,亚群内部的遗传距离为0.040～0.369。推测杏树上的“桃蛀果蛾 ”已经分化成为不同的物种。     

桃小食心虫成虫期高表达气味结合蛋白基因的克隆与表达谱分析

【目的】桃小食心虫Carposina sasakii是我国北方落叶果树的重要蛀果害虫,一旦幼虫蛀入果内,便会对果实的品质产生影响。成虫期是控制此害虫发生为害的关键时期。气味结合蛋白(odorant binding protein, OBP)作为昆虫嗅觉感受系统中与气味分子结合的重要气味运转蛋白,在成虫寄主植物定位及交配行为中具有重要作用。本研究对桃小食心虫气味结合蛋白基因进行克隆、鉴定和成虫组织表达分析,以期为OBPs在桃小食心虫嗅觉感受过程中的功能研究奠定基础。【方法】基于前期获得的桃小食心虫转录组测序数据,选择在其雌雄成虫触角中相对高表达的5个OBP基因(CsasOBP7, CsasOBP12, CsasOBP15, CsasOBP19和CsasOBP21)。采用RACE技术克隆出这5个OBP基因cDNA全长序列,进行生物信息学分析;通过qRT-PCR技术检测这5个OBP基因在桃小食心虫不同发育阶段(1和5日龄卵、初孵幼虫、老熟幼虫和蛹)以及在刚羽化、交配高峰期的和交配后6 h的雌雄成虫不同组织［触角、头(不含触角)、胸、腹、足和翅］中表达量。【结果】获得桃小食心虫5个OBP基因CsasOBP7, CsasOBP12, CsasOBP15, CsasOBP19和CsasOBP21的全长cDNA序列(GenBank登录号: MZ476786-MZ476790)。其中CsasOBP19为一段C端不完整的Minus-C OBP,其余均属于完整的Classical OBPs,且均含有信号肽。系统发育分析表明,在桃小食心虫5个CsasOBPs中, CsasOBP7和CsasOBP19的亲缘关系最近。qRT-PCR结果表明,CsasOBP7和CsasOBP19均在桃小食心虫蛹期高表达,而CsasOBP12, CsasOBP15和CsasOBP21均在卵期高表达。从桃小食心虫成虫刚羽化、交配高峰直至交配后6 h,5个CsasOBP基因表达量总体呈下降趋势。在成虫刚羽化时,这5个CsasOBP基因在各组织中均有表达,且主要在雄虫组织中高表达;在成虫交配高峰期,这5个CsasOBP基因在雄虫触角中高表达,特别是CsasOBP7和CsasOBP15只在雄虫触角中特异性表达;在成虫交配后6 h,每个CsasOBP基因只特异性地高表达于1或2个组织中。【结论】桃小食心虫的这5个CsasOBP基因在成虫交配高峰期雄虫触角中高表达,意味着这些CsasOBP基因可能在雄虫寻找雌虫过程中发挥着重要作用。     

取食不同寄主植物的桃小食心虫飞行能力

【目的】评测取食不同寄主植物的桃小食心虫Carposina sasakii飞行能力,为该虫预测预报和防控提供参考。【方法】使用飞行磨装置测定了取食不同寄主植物(海棠、苹果和杏)的桃小食心虫雌雄成虫的飞行能力,比较了取食同一寄主植物的雌雄成虫间和取食不同寄主植物的同一性别个体间平均飞行距离、平均飞行时间、平均飞行速度和最大飞行速度4个飞行参数。【结果】在12 h的飞行测试中,桃食心虫成虫最长飞行距离为24.54 km,最长飞行时间接近12 h,最大飞行速度为5.88 km/h,飞行11~12h的个体占比最高(36.98%)。取食同一寄主的雌成虫各飞行参数值均大于雄成虫。取食苹果和海棠的成虫的平均飞行距离在雌雄间存在显著差异,取食苹果的雌成虫的平均飞行距离显著大于取食杏的个体,取食苹果的雌成虫平均飞行速度和最大飞行速度显著大于取食另外两种寄主的个体;但是取食不同寄主植物的雄成虫间各飞行参数值差异不显著。【结论】桃小食心虫成虫飞行能力较强,取食同一寄主植物的雌雄成虫间以及取食不同寄主植物的雌成虫间飞行能力存在差异。     

Sex Pheromone Composition and Male Trapping of the Peach Fruit Moth,Carposina sasakii (Matsumura) (Lepidoptera: Carposinidae) in Korea

Sex pheromone composition, mating behavior, and field trapping were analyzed for the peach fruit moth, Carposina sasakii(Matsumura) to develop a monitoring system with its sex pheromone. The upwind flying response of males to female was the most frequent at 4th to 5th hr after lights-off. The highest mating rate was seen at 5th hr after lights-off. The extract of female abdominal tip was analyzed with gas chromatography and proved to have two compounds, (Z)-7-eicosen-11-one and (Z)-7-nonadecen-11-one. In field-trapping, however, a lure containing (Z)-7-eicosen-11-one only showed the highest attractivity.     

Bioinformatic analysis of gene encoding odorant binding protein (OBP) 1, OBP2, and chemosensory proteins in Grapholita molesta

In this study, odorant binding proteins (OBPs) and chemosensory protein (CSP), which are associated with the sensitivity of Grapholita molesta, were comprehensively analysed using bioinformatics. The full-length cDNAs of GmolOBP1, GmolOBP2, and GmolCSP were downloaded and their open reading frames (ORFs) were analysed. Their physicochemical properties were determined and their structures and functions were predicted. Additionally, a phylogenetic tree was constructed to investigate the evolutionary relationships among GmolOBP1, GmolOBP2, GmolCSP, and 14 other insect proteins. GmolOBP1, GmolOBP2 and GmolCSP were composed of 164, 161, and 127 amino acids. GmolOBP1 and GmolOBP2 contained 7 and 6 cysteine residues forming 3 disulphide bonds. The transmembrane, hydrophobic, and signal peptide regions overlapped in GmolOBP1 and GmolCSP and were located in the extracellular environment. GmolCSP showed more coiled coils and a smaller cavity in the three-dimensional structure than GmolOBP1 and GmolOBP2. In the phylogenetic tree, GmolOBP1, GmolOBP2, and GmolCSP were in different clusters or sub-clusters. In conclusion, GmolOBP1 and GmolOBP2 shared some common properties with other OBPs. Additionally, GmolOBP1, GmolOBP2, and GmolCSP may have evolved independently.     

Demography and mass-rearing of Carposina sasakii Matsumura (Lepidoptera: Carposinidae) reared on Golden Delicious and Red Fuji apples in the laboratory

Carposina sasakii Matsumura (Lepidoptera: Carposinidae), is one of the most serious fruit-boring pests in over ten species of fruit trees, and is especially damaging apples in the northern of China. The application of new planting systems, i.e., high-density and dwarfing rootstock orchard systems with mixed apple varieties, makes it important to study the fitness of C. sasakii on these apple varieties to gain fundamental knowledge for use in pest management involving this insect. In this study, life table data of C. sasakii were collected using Golden Delicious and Red Fuji apples as hosts. The egg-larva duration of male C. sasakii reared on Golden Delicious apples (22.81 d) was significantly shorter than that reared on Red Fuji apples (24.27 d). The egg-larva mortality in Golden Delicious apples (59.00%) was lower than that in Red Fuji apples (72.49%). The mortality of the pupal stage, however, was higher in Golden Delicious (10.51%) than in Red Fuji (0%). The total oviposition period (TPOP) on Golden Delicious apples (32.94 d) was significantly shorter than in individuals reared on Red Fuji apples (34.19 d). The intrinsic rate of increase (r = 0.0581 d⁻¹), net reproductive rate (R₀ = 7.57 offspring), and finite rate of increase (λ = 1.0598 d⁻¹) were all higher on Golden Delicious than those on Red Fuji. When the net reproductive rate (R₀) was used, the harvest rate of pupae was higher (0.8678) when reared on Golden Delicious apples than when reared on Red Fuji apples (0.8398). When a large cohort size (n = 200) was used for effective bootstrap sample, the P_E values for C. sasakii reared on Golden Delicious apples and Red Fuji apples were both almost equal to 1. For C. sasakii culturing purposes, Golden Delicious apples would be more productive than Red Fuji.     

Increased trapping efficiency for the peach fruit moth Carposina sasakii (Matsumura) with synthetic sex pheromone

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短时高温对桃小食心虫生长发育与繁殖的影响

【目的】桃小食心虫Carposina sasakii是我国北方落叶果树的重要害虫。本研究旨在探索短时高温对桃小食心虫生长发育和繁殖的影响。【方法】在室内23±1℃、 相对湿度80%±7%和15L∶9D条件下, 测定了桃小食心虫卵、 幼虫、 蛹、 成虫在经历35, 38和41℃高温处理1～4 h后各阶段的发育历期、 存活率和产卵量。【结果】 短时高温对卵的孵化率无明显影响; 经41℃处理后, 初蛀果幼虫(1日龄)的发育历期明显延长, 且存活率显著降低, 3日龄以上的幼虫受到的影响不明显; 11日龄蛹的羽化率在38℃和41℃处理中明显降低, 畸形率也显著升高; 经38℃和41℃处理的成虫存活率降低, 寿命缩短, 产卵量也减少。【结论】短时高温处理对桃小食心虫卵的影响较小, 而对成虫的影响较大。这些结果有助于深入了解该虫在高温季节种群数量变动机制。     

Identity and expression pattern of chemosensory proteins in Heliothis virescens (Lepidoptera, Noctuidae)

Analyzing the chemosensory organs of the moth Heliothis virescens, three proteins belonging to the family of insect chemosensory proteins (CSPs) have been cloned; they are called HvirCSP1, HvirCSP2 and HvirCSP3. The HvirCSPs show about 50% identity between each other and 30–76% identity to CSPs from other species. Overall, they are rather hydrophilic proteins but include a conserved hydrophobic motif. Tissue distribution and temporal expression pattern during the last pupal stages were assessed by Northern blots. HvirCSP mRNAs were detected in various parts of the adult body with a particular high expression level in legs. The expression of HvirCSP1 in legs started early during adult development, in parallel with the appearance of the cuticle. HvirCSP1 mRNA was detectable five days before eclosion (day E-5), increased dramatically on day E-3 and remained at high level into adult life. The tissue distribution and the time course of appearance of HvirCSPs are in agreement with a possible role in contact chemosensation.     

Identity and expression pattern of chemosensory proteins in Heliothis virescens (Lepidoptera, Noctuidae)

Analyzing the chemosensory organs of the moth Heliothis virescens, three proteins belonging to the family of insect chemosensory proteins (CSPs) have been cloned; they are called HvirCSP1, HvirCSP2 and HvirCSP3. The HvirCSPs show about 50% identity between each other and 30–76% identity to CSPs from other species. Overall, they are rather hydrophilic proteins but include a conserved hydrophobic motif. Tissue distribution and temporal expression pattern during the last pupal stages were assessed by Northern blots. HvirCSP mRNAs were detected in various parts of the adult body with a particular high expression level in legs. The expression of HvirCSP1 in legs started early during adult development, in parallel with the appearance of the cuticle. HvirCSP1 mRNA was detectable five days before eclosion (day E-5), increased dramatically on day E-3 and remained at high level into adult life. The tissue distribution and the time course of appearance of HvirCSPs are in agreement with a possible role in contact chemosensation.     

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桃小食心虫幼虫的消化道和马氏管的显微结构观察

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Molecular and functional characterization of three odorant binding proteins from the oriental fruit moth Grapholita molesta (Busck) (Lepidoptera: Tortricide)

Odorant binding proteins (OBPs) act in recognizing odor molecules and their most well‐studied functions are transporting odors across the sensillum lymph to olfactory receptor neurons within the insect antennal sensillum. The adults of Grapholita molesta highly depend on olfactory cues in locating host plants and selecting oviposition sites, in which OBPs play an important role in perceiving and recognizing host plant volatiles. Exploring the physiological function of OBPs could facilitate our understanding of their importance in insects’ chemical communication. In this study, three OBP genes were cloned and named GmolOBP4, GmolOBP5, and GmolOBP10. Quantitative real‐time PCR results indicated that GmolOBP4 and GmolOBP10 were predominantly expressed in adult antennae and GmolOBP5 was expressed in multiple tissues, including head, legs, and wings in addition to antennae. The binding affinities of the three recombinant GmolOBPs (rGmolOBPs) with four sex pheromone components and twenty‐nine host plant volatiles were measured using 1‐N‐Phenyl‐naphthylamine as a fluorescence probe. The three rGmolOBPs exhibited specific binding properties to potential ligands, GmolOBP4 and GmolOBP10 bound to minor sex pheromone components, such as (Z)‐8‐dodecenyl alcohol and dodecanol, respectively. rGmolOBP4 showed intermediate binding ability with hexanal, benzyl alcohol, and pear ester, rGmolOBP5 had a weak affinity for benzaldehyde, pear ester and, methyl jasmonate, and rGmolOBP10 showed strong binding capacity toward hexanol, decanol, and α‐ocimene. We speculate that the GmolOBP4 and GmolOBP10 have dual functions in perception and recognition of host plant volatiles and sex pheromone components, while GmolOBP5 may serve other function(s).