Localization of RR-1 and RR-2 cuticular proteins within the cuticle of Anopheles gambiae

The largest arthropod cuticular protein family, CPR, has the Rebers and Riddiford (R&R) Consensus that in an extended form confers chitin-binding properties. Two forms of the Consensus, RR-1 and RR-2, have been recognized and initial data suggested that the RR-1 and RR-2 proteins were present in different regions within the cuticle itself. Thus, RR-2 proteins would contribute to exocuticle that becomes sclerotized, while RR-1s would be found in endocuticle that remains soft. An alternative, and more common, suggestion is that RR-1 proteins are used for soft, flexible cuticles such as intersegmental membranes, while RR-2s are associated with hard cuticle such as sclerites and head capsules. We used TEM immunogold detection to localize the position of several RR-1 and RR-2 proteins in the cuticle of Anopheles gambiae. RR-1s were localized in the procuticle of the soft intersegmental membrane except for one protein found in the endocuticle of hard cuticle. RR-2s were consistently found in hard cuticle and not in flexible cuticle. All RR-2 antibodies localized to the exocuticle and four out of six were also found in the endocuticle. Hence the location of RR-1s and RR-2s depends more on properties of individual proteins than on either hypothesis.     

Molecular Characterization and Imidacloprid Selectivity of Nicotinic Acetylcholine Receptor Subunits from the Peach-Potato Aphid Myzus persicae

The recent introduction of the chloronicotinyl insecticide imidacloprid, targeting insect nicotinic acetylcholine receptors (nAChRs), emphasises the importance of a detailed molecular characterisation of these receptors. We are investigating the molecular diversity of insect nAChR subunit genes in an important agricultural pest, the peach-potato aphid Myzus persicae. Two M. persicae alpha-subunit cDNAs, Mp alpha1 and Mp alpha2, have been cloned previously. Here we report the isolation of three novel alpha-subunit genes (Mp alpha3-5) with overall amino acid sequence identities between 43 and 76% to characterised insect nAChR subunits. Alignment of their amino acid sequences with other invertebrate and vertebrate nAChR subunits suggests that the insect alpha subunits evolved in parallel to the vertebrate neuronal nAChRs and that the insect non-alpha subunits are clearly different from vertebrate neuronal beta and muscle non-alpha subunits. The discovery of novel subtypes in M. persicae is a further indicator of the complexity of the insect nAChR gene family. Heterologous co-expression of M. persicae nAChR alpha-subunit cDNAs with the rat beta2 in Drosophila S2 cells resulted in high-affinity binding of nicotinic radioligands. The affinity of recombinant nAChRs for [3H]imidacloprid was influenced strongly by the alpha subtype. This is the first demonstration that imidacloprid selectively acts on Mp alpha2 and Mp alpha3 subunits, but not Mp alpha1, in M. persicae.     

Characterization of cuticular proteins from the migratory locust,Locusta migratoria

Proteins were extracted from the still unhardened (teneral) cuticle of the migratory locust, Locusta migratoria. The proteins are soluble only at extreme pH-values and at low ionic strength, the solubility increases with decreasing temperature. The unhardened cuticle contains approx. 100 different proteins according to two-dimensional polyacrylamide gel electrophoresis. The majority of the proteins are very basic. The basicity and solubility properties of the proteins have necessitated development of modified electrophoretic procedures. The amino acid composition of the bulk protein shows that alanine, proline, glycine, valine and tyrosine constitute two thirds of the total amino acid content and that cysteine, methionine and tryptophan are absent.The proteins have been extracted from various parts of the cuticle and analysed by two-dimensional electrophoresis. Characteristic protein compositions were found for cuticle from the different body parts. Amino acid analyses of these extracts are strikingly similar. The only significant difference is in the glycine-alanine ratio. Cuticles that are destined to become hard are extremely rich in alanine, whereas the flexible parts of the cuticle are enriched in glycine. The results indicate that the proteins of locust cuticle constitute a group of structural proteins different from other known structural proteins.     

Characterization of a new densovirus infecting the green peach aphid Myzus persicae

A new icosahedral DNA virus was isolated from aphids (Myzus persicae) that showed abnormal growth and development. The purified virus particles have a diameter of 20 nm and contain a single-stranded DNA molecule of approximately 5.7 kb. The viral particles are composed of five structural proteins (92, 85, 68, 64, and 57 kDa). As the main biophysical properties of this virus are similar to those of the members of the genus Densovirus it was tentatively named Myzus persicae densovirus (MpDNV). A PCR-based detection method and a polyclonal antiserum raised against MpDNV allowed the detection of the virus in a single-infected aphid. MpDNV is immunologically related to Junonia coenia densovirus, but not to other members of the subfamily Densovirinae. Biological assays showed that MpDNV could be both transmitted transovarially and horizontally via honeydew and saliva. MpDNV was able to infect whiteflies but not other aphid species tested.     

桃蚜体内与病毒结合的共生菌Buchnera groEL基因的克隆和序列分析

以桃蚜(Myzus persicae)杨凌生物型为材料,利用一对特异性引物,用PCR的方法从桃蚜体内扩增出了内共生菌的Buchnera groEL基因,序列测定结果表明其长度为1 647bp,与GenBank中的桃蚜荷兰生物型、蜿豆蚜(A-cyrthosi phonpisum)日本生物型Buchnera GroEL基因长度相同,同源性分别为99%和91%;Buchnera GroEL-YL编码548个氨基酸,序列比较发现与Buchnera GroEL-NT仅有3个氨基酸的差异,即AA111Met→Lys、AA222Val→Met and AA348G1n→His.     

Genetic, biochemical, and behavioral uniformity among populations of Myzus nicotianae and Myzus persicae

Prior to designation as distinct species, an appellation presently in question, the tobacco aphid, Myzus nicotianae Blackman (Homoptera: Aphididae), was classified as a tobacco-feeding form of the green peach aphid, Myzus persicae (Sulzer). In this study, RAPD polymorphisms distinguished members of the Myzus persicae complex (M. persicae and M. nicotianae) from three outgroup Myzus species (M. cerasi (F.), M. hemerocallis Takahashi, and M. varians Davidson). Polymorphisms within the complex did not separate populations on the basis of host association (tobacco versus other host plants) or geographic origin (collections from the United States, Europe, and Japan). Similarly, while GC-MS analysis of cuticular hydrocarbon profiles revealed both developmental and inter-populational differences within the M. persicae complex, it did not separate populations of tobacco feeding aphids from those collected off non-tobacco hosts. Finally, with the exception of their responses to a choice between lettuce and collards, the host preference behavior of a green peach aphid population, a red tobacco aphid population, and a green tobacco aphid population was indistinguishable in host preference experiments. These results add to a growing body of evidence suggesting M. nicotianae and M. persicae are conspecific.     

七星瓢虫成虫对烟蚜的捕食作用

对七星瓢虫CoccinellaseptempunctataL .成虫对烟蚜Myzuspersicae的捕食作用进行了定量研究。七星瓢虫成虫对烟蚜的功能反应属HollingⅡ型反应 ,拟合的圆盘方程为Na =1 .1 5 76Nt ( 1 +0 .0 0 3 48Nt) ,χ2 检验表明圆盘方程理论值与实测值相符。捕食选择试验表明在烟蚜、烟青虫Helicoverpaassaut(Guenee)卵和 1龄幼虫 3种猎物中 ,七星瓢虫成虫最喜好烟蚜。七星瓢虫成虫对自身密度的功能反应用Watt模型拟合为A =2 90 .0 3P-0 .7584,其捕食作用率与个体间相互干扰作用的关系用Hassell模型拟合为E =0 .8783 7P-0 .1 0 0 94。文中进一步就烟田中七星瓢虫的保护利用进行了讨论。     

Feeding behavior of Myzus persicae on asparagus species susceptible and resistant to Asparagus virus 1

Asparagus virus 1 (AV‐1) infects Asparagus officinalis L. (Asparagaceae) in the field worldwide. However, various wild relatives of A. officinalis are resistant to AV‐1. Here we study the behavior of the green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae), on 19 AV‐1‐resistant wild relatives of A. officinalis. We focus on behavior that is associated with regular cell penetration, relevant for inoculation of AV‐1, and sieve element penetration to check for vector resistance and its potential influence on AV‐1 transmission. Parameters, relevant for the transmission of non‐persistent viruses and host plant acceptance, were obtained by the electrical penetration graph technique. Furthermore, phylloclade architecture of A. officinalis and its wild relatives was examined to study its influence on aphid behavior. Behavior of M. persicae displays many cell penetrations and long ingestion periods on A. officinalis, compared to the generally shorter cell penetrations (reduced potential for virus transmission) and reduced or no ingestion (phloem‐located aphid resistance) on wild relatives. Because effects on aphid behavior are not consistent throughout the group of the tested wild relatives of A. officinalis, with some wild relatives being susceptible to M. persicae, a common genetic background for AV‐1 and aphid resistance appears to be unlikely. However, the reduced potential of virus transmission as well as aphid resistance shown by some wild relatives may be useful for future breeding programs.     

大草蛉对桃蚜和夹竹桃蚜的捕食作用研究

大草蛉在我国分布广泛,能捕食多种农作物害虫。本试验首先研究了大草蛉一龄幼虫对桃蚜、大草蛉二龄幼虫对桃蚜和夹竹桃蚜的捕食功能反应。三个反应的最大捕食量Namax分别为144．93、303．03和151．51（头）,瞬时攻击率a分别为0．7428、0．8654和0．6644。大草蛉一龄幼虫有较强的种内干扰反应,随着捕食者密度的增加,捕食作用率（E）依次为0．2025、0．1185、0．0866、0．0694和0．0584;对桃蚜的捕食作用率（E）与自身密度（P）的函数关系式为E=0．2025P^-0.7727。在桃蚜和夹竹桃蚜的混合种群中,大草蛉二龄幼虫对桃蚜的选择效应大于夹竹桃蚜。     

桃蚜MpAChE基因RNAi表达载体构建及转化

通过害虫取食植物表达害虫发育关键基因dsRNA的转基因植株,分析能否通过抑制害虫特定基因的表达来防控害虫。本研究利用RT-PCR技术从桃蚜中克隆乙酰胆碱酯酶基因383 bp cDNA片段,命名为MpAChE。进一步利用该MpAChE基因片段构建植物RNAi表达载体RNAi-MpAChE,并通过浸花法转化野生型拟南芥,通过卡那霉素抗性筛选转化植株,PCR及Southern杂交进一步鉴定转基因植株。结果表明:克隆的cDNA片段与桃蚜中已克隆的乙酰胆碱酯酶(GenBank登录号AY147797)cDNA序列核苷酸一致性为99%。卡那霉素抗性初步筛选和PCR进一步鉴定,获得25株阳性转基因植株。从25株中随机选择的5株阳性植株,Southern杂交均为阳性。经接种桃蚜初步鉴定,转基因植株对蚜虫的抗性效果不显著。     

“秦油2号”油菜植株内含物与桃蚜种群消长的关系

通过对N、P、K不同施肥条件下的“秦油2号”油菜植株内含物与桃蚜种群消长关系的研究与分析,建立了9个数学模型,结果表明可溶性糖含量、含K量、含水量对桃蚜种群消长影响最大;低N、低K、中P组合对桃蚜种群增长不利.     

桃蚜体内与病毒结合的共生菌Buchnera groEL基因的克隆和序列分析

以桃蚜（Myzus persicae)杨凌生物型为材料,利用一对特异性引物,用PCR的方法从桃蚜体内扩增出了内共生菌的Buchnera groEL基因,序列测定结果表明其长度为1647bp,与GenBank中的桃蚜荷兰生物型、蜿豆蚜（A-cyrthosiphon pisum)日本生物型Buchnera GroEL基因长度相同,同源性分别为99%和91%;Buchnera GroEL-YL编码548个氨基酸,序列比较发现与BuchneraGroEL-NT仅有3个氨基酸的差异,即AA111Met→Lys,AA222Val→MetandAA348Gln→His。     

Supercooling of Myzus persicae in relation to gut content

The supercooling distribution curves of Myzus persicae varied in response to feeding position, diet and age. Unfed instar I aphids supercooled to temperatures lower than –20 °C. Feeding aphids supercooled to higher temperatures, the earlier instars on subsidiary veins supercooling to lower temperatures than the late instars and adults on the main veins. Starvation decreased supercooling ability in some adult aphids. The supercooling ability of aphids feeding on sucrose solutions through a Parafilm membrane decreased in response to an increasing concentration, the position being reversed on 25% sucrose. Varying the amino acid concentration had no effect. The possible effects of this variable supercooling ability on the winter survival of the aphid are discussed.     

Low temperature mortality of the peach-potato aphid Myzus persicae

ABSTRACT.

• 1 The mean supercooling points of first instar and adult Myzus persicae (Sulzer) maintained at 20°C and cooled at 1°C min^?1 were ?26.6 and ?25.0°C respectively.
• 2 The LT₅₀ (temperature) of the same age groups drawn from the same population and cooled at the same rate were ?8.1 and ?6.9°C, indicating extensive pre-freeze mortality in M.persicae under laboratory conditions.
• 3 Acclimation at 10 and 5°C did not affect supercooling but depressed the LT₅₀ of both first instars and adult aphids.
• 4 Freezing of leaves during feeding did not increase mortality above that expected from the direct effects of low temperature.
• 5 The level of cold in different winters can be expressed in terms of the total number of frost days, and the frequency of abnormally cold days. Winter temperatures differ markedly in a vertical profile from the soil to the soil or grass surface, and then to the air (and foliage) above.
• 6 The time of the first record of M.persicae in suction trap samples is correlated with January and February temperatures except in the west of England and Wales. Further north December and January temperatures are relatively more important.
• 7 Winter temperatures and the resultant aphid mortality is a primary determinant of the timing of the spring migration.

     

Reproduction and feeding behavior of Myzus persicae on four cereals

Green peach aphid, Myzus persicae (Sulzer), does not overwinter outdoors in Minnesota; it arrives each spring on low-level jet streams from the south. After arrival, anholocylic reproduction occurs on numerous herbaceous species, including many common weeds, before movement to potato, Solanum tuberosum L. In investigating aphid feeding behavior on barrier crops, we observed winter wheat, Triticum aestivum L., colonized by green peach aphid. The Northern Great Plains grows 94,000 ha of potatoes and 20.5 million ha of small grain cereals each year, the latter potentially providing an early emerging and widely distributed green peach aphid host to influence early season potato colonization. Life tables statistics indicated green peach aphid had its highest reproductive potential among cereals on winter wheat, with rye (Secale cereale L.) > barley (Hordeum vulgare L.) > oats (Avena sativa L.). Green peach aphid was found to colonize barley, rye, and winter wheat, but not oats. Mean generation time, net reproductive rate, doubling time, and finite rate of increase were significantly different between host plants. Electrical penetration graph technique indicated mean nonpenetration duration by green peach aphid was significantly different among plant species, and significantly longer on winter wheat than on the other cereals. Mean xylem phase duration was not significantly different among plant species but sieve element salivation was of longest duration on potato. Phloem sap ingestion (E2) was also significantly different among plant species with longest E2 duration on winter wheat. This study demonstrates that this aphid can effectively use key cereals at the vegetative stage.     

Evaluating reproductive fitness and metabolic costs for insecticide resistance in Myzus persicae from Chile

The development of insecticide resistance in pest insects is an increasing problem for agriculture, forestry and public health. Aphids are ubiquitous herbivorous insects, with approximately 4700 known species, of which less than 5% exploit the agricultural environment successfully. Of these, the peach‐potato aphid Myzus persicae Sulzer is recognized as one of the most important pests worldwide because it has acquired resistance to many insecticides. Although resistance to insecticides provides important benefits for pests in agricultural fields that are treated with insecticides, it may be associated with fitness (or other) costs in environments that are insecticide free. In the present study, the fitness and energy costs that might be experienced by M. persicae in an insecticide‐free environment when carrying at least one insecticide resistance mutation (IRM), or by having an increased production of esterases, are evaluated. The study investigates whether genotypes that have an IRM also have enhanced esterase production, whether there is any metabolic cost associated with insecticide resistance, and whether there are any fitness costs associated with insecticide resistance and metabolic expenditure. The intrinsic rate of increase, standard metabolic rate (i.e. a measure of maintenance costs) and constitutive esterase activity are determined for 30 different multilocus genotypes carrying (or not carrying) at least one of the two most frequent insecticide resistance mutations (MACE and kdr/super‐kdr) that occur in Chile. The results show that genotypes carrying at least one IRM have higher levels of total esterase activity than genotypes without an IRM, that there is no evidence of an energy cost associated with total esterase activity or IRM, and no evidence for a reproductive fitness cost associated with total esterase activity, IRM or metabolic rate. The results agree with previous studies showing linkage disequilibrium between insecticide resistance mechanisms, although they contrast with those of studies that report fitness costs associated with insecticide resistance in Myzus persicae.     

Gustation of dietary amino acids by the aphid Myzus persicae

In simple choice-chamber tests the ability of the aphid Myzus persicae (Sulzer) to discriminate between sucrose solutions containing single or groups of amino acids and a pure sucrose solution was assessed. Each of 20 dietary amino acids was tested at a concentration of 0.1% in 20% sucrose. Each of six amino acids (asparagine, isoleucine, leucine, methionine, phenylalanine, and tryptophan) strongly enhanced the acceptability of sucrose. Methionine had a strong effect also at 0.02%. A few other amino acids had a lesser effect, while only a slight or no effect was observed with about half of the 20 dietary amino acids. A slightly deterrent effect was observed with histidine, glutamic acid, and arginine.The experimental apterous adult aphids that had been reared on radish seedlings had to be preconditioned on a pure sucrose solution for one day prior to the tests in order to show these responses within a few hours.

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Zusammenfassung In einfachen Wahlkammer-Versuchen wurde die Fähigkeit der Blattlaus Myzus persicae geprüft, Rohrzuckerlösungen, die einzelne oder Gruppen von Aminosäuren enthielten, von reiner Rohrzuckerlösung zu unterscheiden. Jede der 20 diätischen Aminosäuren wurde in einer Konzentration von 0,1% in 20% iger Rohrzuckerlösung geprüft. Jede der 6 Aminosäuren Asparagin, Isoleucin, Leucin, Methionin, Phenylalanin und Tryptophan erhöhte die Annehmbarkeit stark. Unter ihnen hatte Methionin auch bei 0,02% eine kräftige Wirkung. Einige wenige andere Aminosäuren hatten einen geringen Effekt, während bei etwa der Hälfte der 20 diätischen Aminosäuren nur ein schwacher oder kein Einfluß beobachtet wurde. Eine leicht abweisende Wirkung war bei Histidin, Glutaminsäure und Arginin festzustellen.Die im Versuch verwendeten erwachsenen apteren Blattläuse, die an Rettichsämlingen herangezogen worden waren, mußten vor den Versuchen einen Tag lang an reiner Rohrzuckerlösung gehalten werden, wenn sie diese Reaktionen innerhalb einiger weniger Stunden zeigen sollten.

     

不同桃树品种上桃蚜遗传多样性的研究

桃蚜Myzus persicae(Sulzer)寄主广泛,是一种全球性的广食性害虫。为了探明不同桃树品种对桃蚜遗传分化的影响,我们采用微卫星分子标记技术对白油蟠桃,黄油蟠桃等7个桃树品种上的桃蚜进行遗传多样性研究。结果表明,7对微卫星引物在171个样本中检测到118个等位基因,平均每个位点有16.857个等位基因。不同品种桃树上桃蚜的遗传多样性较为丰富,表明种内存在着较大的遗传变异。种群间没有遗传分化或分化不明显,这可能是种群间存在着明显的基因交流而造成的,说明桃树品种间的差异对桃蚜的分化影响较小。