Sampling of Cacopsylla pyri (Hemiptera: Psyllidae) and Pilophorus gallicus (Hemiptera: Miridae) in pear orchards

The psyllid Cacopsylla pyri L. (Hemiptera: Psyllidae), is one of the principal pests of pear (Pyrus spp.) orchards and, along with its natural enemies, needs to be carefully monitored for correct integrate pest management and biological pest control decision making. We compare sampling techniques and develop sampling methods for C. pyri and Pilophorus gallicus Remane (Heteroptera: Miridae). Four pear orchards were sampled periodically from 2007 to 2010 in southern Spain by beating branches into funnels or 45-cm-diameter nets and taking shoot samples. Different sampling techniques were compared, and several sampling methods were assayed for adults and nymphs of C. pyri and P. gallicus. Psylla adult and P. gallicus counts in the nets and funnels were similar and closely correlated. Counts of C. pyri adults in the funnels and nets were higher than on shoots, but all measures were closely correlated. The number of nymphs on shoots and leaves was correlated, as were the counts on leaves within shoots. A guide for choosing the sampling unit in cost/precision terms is given. C. pyri nymphs and adults showed an aggregated distribution, whereas P. gallicus had a less-crowded spatial distribution. Enumerative sequential sampling was evaluated for both insects by using the different sampling techniques by resampling experimental data. The negative binomial distribution fitted the experimental data gathered using the different sampling techniques for C. pyri adults and P. gallicus. The use of binomial sampling is discussed for the two insects and the costs involved are compared with those implied in the sequential enumerative method.     

明脉喀木虱Cacopsylla hyalinonemae Li&Yang,1989雄性首次描述(半翅目:木虱总科:木虱科)

首次描述了明脉喀木虱Cacopsylla hyalinonemae LiYang的雄性,对雌性进行了详细的重新描述,并给出了本种的主要特征图,背面、侧面以及头部正面的形态照片。标本保存于贵州大学昆虫研究所。     

Seasonal variation in susceptibility of the pear psyllid,Cacopsylla permixta (Hemiptera: Psyllidae) to selected insecticides

Although Cacopsylla pyri Forster and Cacopsylla pyricola Linnaeus have long been considered as more significant pests of pear trees around the world, Cacopsylla permixta Burckhardt and Hodkinson is the most significant pest in some parts of Iran, especially in pear gardens of Karaj. Current control strategies against this pest in Iran generally involve five or six insecticide treatments each year, despite unsatisfactory results are reported at many localities. So, it is crucial to know the most susceptible generation of the pest to apply a good strategy for its control. The aim of this study was to explore the seasonal variation in susceptibility of C. permixta to four commonly used insecticides. The results showed that winter forms (February) were 2.71‐fold, 4.58‐fold, 3.26 fold and 3.38‐fold more tolerant to diazinon, imidacloprid, acetamiprid and abamectin, respectively, compared with summer forms. Also, Esterase, GST and P450 monooxygenase activity was highest during February. Moreover, the content of lipid, carbohydrate, glycogen and protein was significantly higher in February compared with other months. Based on these results, the best period for insecticide treatment for efficient control is treatment against the first generation, at the time when eggs are laid by females as well as during the egg hatching and the larvae appearance. At this time of year, psylla are more susceptible, which would likely lead to better results and the reduction in damage during the next summer. However, further studies are needed to test this in farm settings, and to whether this holds true for other psylla species.     

微卫星标记分析中国梨木虱种群的遗传多样性

中国梨木虱Cacopsylla chinensis (Yang et Li)是梨树主要害虫之一。为了从分子水平评估中国梨木虱种群内的遗传变异和种群间的遗传分化, 本文应用7对微卫星DNA引物对中国16个地区中国梨木虱种群进行遗传多样性分析。结果表明： 各位点有效等位基因为2.2927～10.0610, 多态信息含量(PIC)值在0.5073～0.8735之间。16个种群的平均期望杂合度为0.7876, 遗传距离在0.0951～1.0139之间, Nei氏期望杂合度为0.4771～0.7892, Shannon信息指数在0.8396～1.9989之间, 群体分化率F_ST为11.61%, 基因流平均值为2.2236。结果表明, 7个微卫星位点均具有较高的多态性, 各种群间的遗传分化水平较低, 基因交流程度较高, 遗传变异主要存在于种群内的个体间。研究结果为制定针对中国梨木虱的有效防治策略提供部分分子生物学的基础资料。     

Ecological and Genetic Differences between Cacopsylla melanoneura (Hemiptera,Psyllidae) Populations Reveal Species Host Plant Preference

The psyllid Cacopsylla melanoneura is considered one of the vectors of ‘Candidatus Phytoplasma mali’, the causal agent of apple proliferation disease. In Northern Italy, overwintered C. melanoneura adults reach apple and hawthorn around the end of January. Nymph development takes place between March and the end of April. The new generation adults migrate onto conifers around mid-June and come back to the host plant species after overwintering. In this study we investigated behavioural differences, genetic differentiation and gene flow between samples of C. melanoneura collected from the two different host plants. Further analyses were performed on some samples collected from conifers. To assess the ecological differences, host-switching experiments were conducted on C. melanoneura samples collected from apple and hawthorn. Furthermore, the genetic structure of the samples was studied by genotyping microsatellite markers. The examined C. melanoneura samples performed better on their native host plant species. This was verified in terms of oviposition and development of the offspring. Data resulting from microsatellite analysis indicated a low, but statistically significant difference between collected-from-apple and hawthorn samples. In conclusion, both ecological and genetic results indicate a differentiation between C. melanoneura samples associated with the two host plants.     

The complete mitochondrial genome of Diaphorina citri (Hemiptera: Psyllidae) and phylogenetic analysis

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is the most serious pest of citrus as the vector of Huanglongbing (HLB), the citrus greening disease. In this study, the complete mitochondrial genome (mitogenome) of D. citri has been sequenced and annotated, and a comparative analysis is provided with known Psylloidea species. The mitogenome of D. citri is a typical circular molecular of 15,038 bp in length with an A + T content of 74.56%, contains the typical 37 genes and the gene order is identical to the other Psylloidea mitogenomes. The nucleotide composition and codon usage of D. citri are similar to the four Psylloidea species. All protein-coding genes (PCGs) use standard initiation codons (TAN), stop with TAA and TAG except ND2 and ND5 which stop with incomplete termination codon T. All tRNAs have the typical clover-leaf structure, with the exception of trnS1 lacking the dihydrouridine (DHU) arm. The control region is located between rrnL and the trnI gene with the highest A + T content among the five Psylloidea species. Phylogenetic analysis is conducted based on the 13 PCGs or/and 2 rRNAs of 23 Sternorrhycha mitogenomes. Both maximum likelihood (ML) and Bayesian inference (BI) analysis suggest a clear relationship of Psylloidea, Aleyrodoidea and Aphidoidea within Sternorrhycha, which support the traditional morphological classification.     

Identification of two invasive Cacopsylla chinensis (Hemiptera: Psyllidae) lineages based on two mitochondrial sequences and restriction fragment length polymorphism of cytochrome oxidase I amplicon

The occurrence of pear decline, a disease found in some pear (Pyrus spp.) orchards of Taiwan in recent years, is accompanied by an outbreak of Cacopsylla chinensis (Yang & Li). Two major morphological forms (summer and winter forms) with a variety of intermediate body color and two phylogenetic lineages of this psyllid have been described. The work herein used sequences of mitochondrial cytochrome oxidase I (COI) and 16S rDNA regions to delineate the genetic differentiation of this color-variable insect and to elucidate their relationship. Sequence divergence and phylogenetic analysis have shown that C. chinensis individuals could be divided into two lineages with 3.3 and 2.3% divergence of COI and 16S rDNA, respectively. All specimens from China were found to belong to lineage I. Restriction fragment length polymorphism analysis of COI with restriction enzymes AcuI, AseI, BccI, and FokI on 263 specimens of six populations from Taiwan produced two digestion patterns, which are in agreement with the two lineages described above. Both patterns could be found in each population, with most individuals belonging to lineage I and 5-21% of the individuals belonging to lineage II. Because these two lineages included summer as well as winter morphological forms, the lineage differentiation is apparently not related to morphological characters of this psyllid. Because the invasive records are not in favor of a sympatric differentiation, this psyllid is more likely introduced as different populations from countries in temperate regions.     

Transmission of apple proliferation phytoplasma by Cacopsylla melanoneura (Homoptera: Psyllidae)

Transmission trials of apple proliferation (AP) phytoplasma to healthy apple plants were carried out with Cacopsylla melanoneura (Forster). Both field naturally infected and experimentally infected psyllids were evaluated. The capacity of the different life stages of the insect in transmitting AP was tested. Overwintered adults collected in the orchards were able to transmit AP with a variable efficiency, depending on the infectivity rate of source plants. Experimentally infected nymphs and springtime adults succeeded in the transmission of AP, but the lower number of insect tested and the shorter inoculation period, due to difficulties in rearing the whole cycle of the insect in the laboratory, affected the efficiency. Considering the life history of C. melanoneura, the overwintered adults are the most responsible of the diffusion of AP in apple orchards.     

Global relationships of Bemisia tabaci (Hemiptera: Aleyrodidae) revealed using Bayesian analysis of mitochondrial COI DNA sequences

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a species complex that is one of the most devastating agricultural pests worldwide and affects a broad range of food, fiber and ornamental crops. Unfortunately, using parsimony and neighbor joining methods, global phylogenetic relationships of the major races/biotypes of B. tabaci remain unresolved. Aside from the limitations of these methods, phylogenetic analyses have been limited to only small subsets of the global collection of B. tabaci, and thus limited taxon sampling has confounded the analyses. To improve our understanding of global B. tabaci phylogenetic relationships, a Bayesian phylogenetic technique was utilized to elucidate the relationships among all COI DNA sequence data available in GenBank for B. tabaci worldwide (366 specimens). As a result, the first well-resolved phylogeny for the B. tabaci species complex was produced showing 12 major well-resolved (0.70 posterior probability or above) genetic groups: B. tabaci (Mediterranean/Asia Minor/Africa), B. tabaci (Mediterranean), B. tabaci (Indian Ocean), B. tabaci (sub-Saharan Africa silverleafing), B. tabaci (Asia I), B. tabaci (Australia), B. tabaci (China), B. tabaci (Asia II), B. tabaci (Italy), B. tabaci (New World), B. tabaci (sub-Saharan Africa non-silverleafing) and B. tabaci (Uganda sweet potato). Further analysis of this phylogeny shows a close relationship of the New World B. tabaci with Asian biotypes, and characteristics of the major sub-Saharan Africa non-silverleafing clade strongly supports an African origin of B. tabaci due to its position at the base of the global phylogeny, and the diversity of well-resolved sub-clades within this group. Bayesian re-analyses of B. tabaci ITS, COI, and a combined dataset from a previous study resulted in seven major well-resolved races with high posterior probabilities, also showing the utility of the Bayesian method. Relationships of the 12 major B. tabaci genetic groups are discussed herein.     

Chromosome banding in Cacopsylla mali (Schmidberger) and Cacopsylla sorbi (Linnaeus) (Homoptera, Psyllidae) with polymorphic sex chromosomes

C-banding and Ag staining were applied to Cacopsylla sorbi and C. mali. The aim was to discover some additional cytological markers to follow the pathways of the sex determination system transformation in the evolution of the species. Of three karyotype patterns so far described in the literature for these species--a type A (XO), B (neoXY) and C (neoX1X2Y), only the last two types (B and C) were found. All 6 studied Cacopsylla sorbi from Finland had a karyotype of type B (2n = 20 + XY), while C. mali had both types. Type B (2n = 22 + XY) was observed in 31 males, whereas type C (2n = 20 + neoX1X2Y) in the remaining four. The karyotype of C. sorbi was found to be characterized by a very small amount of C-positive material, localized in a telomere of the Y chromosome. The karyotype of C. mali was also characterized by a very small amount of C-banded material. Both the sex chromosomes and the autosomes displayed a marked polymorphism of C-positive bands within different individuals and even the same individual. In both species the nucleolus was located in the telomere of a middle sized autosomal till diplotene inclusive. The C-banding and Ag staining in the studied Cacopsylla species did not provide any additional cytological markers for an understanding of the pathways of sex determination system transformation in the evolution of the species.     

Population dynamics of Cacopsylla melanoneura (Hemiptera: Psyllidae) in northeast Italy and its role in the apple proliferation epidemiology in apple orchards

In the current study, incidence of 'Candidatus Phytoplasma mali' in an experimental apple orchard in northeast Italy, in addition to abundance and phytoplasma infectivity of Cacopsylla melanoneura (F?rster) (Hemiptera: Psyllidae) was determined and the role of this psyllid as a vector of 'Ca. P. mali' in this region was reviewed. Insect samples collected in the orchard by the beating method indicated high abundance of C. melanoneura (up to 7.92 specimens/branch); however, the psyllid C. picta was not observed. Molecular analyses revealed presence of 'Ca. P. mali' in 6.25% of overwintered psyllids. This infection rate is quite high in comparison to other localities where C. melanoneura is known as the main vector of the phytoplasma. This finding supports the assumption that C. melanoneura also is paramount in the epidemiology of the apple proliferation disease also in northeast Italy. Moreover, we correlated immigration dynamics to the temperatures registered in the apple orchard, and defined an immigration index to predict the progressive arrival of the overwintered adults from winter sites. Psyllids start to reach the apple orchards when either the average of the maximum temperature of the 7 d is above 9.5 degrees C or the immigration index has a positive value. This index will be a useful tool for the growers to prevent apple proliferation phytoplasma spread with well-timed insecticide treatments targeted against C. melanoneura. However, further research is needed to validate or adjust the index to other apple growing regions, which may affect more efficacious management of this disease and psyllid vector.     

Fine structure and sensory apparatus of the mouthparts of the pear psyllid,Cacopsylla chinensis (Yang et Li) (Hemiptera: Psyllidae)

The pear psyllid, Cacopsylla chinensis (Yang et Li) (Hemiptera: Psyllidae), is one of the most significant economic pests of pear in China, causing direct damage through feeding by the highly specialized piercing–sucking mouthparts. The ultrastructural morphology and sensory apparatus of the mouthparts of the adult were examined using scanning and transmission electron microscopy. The piercing–sucking mouthparts of C. chinensis are composed of a three-segmented labium with a deep groove in the anterior side, a stylet fascicle consisting of two mandibular and two maxillary stylets, and a pyramid-shaped labrum. Proximal to the labium, the stylet fascicle forms a large loop within a membranous crumena. Mandibles, with more than ten teeth on the external convex region, can be seen on the distal extremity. Smooth maxillary stylets are interlocked to form a larger food canal and a smaller salivary canal. One dendritic canal housing 2 dendrites is also found in each mandible. Two types of sensilla trichodea, four types of sensilla basiconica, single as well as groups of sensilla campaniformia, and oval flattened sensilla occur in different locations on the labium, whereas a kind of sensilla basiconica is at the junction of the labrum and anteclypeus. Sensilla trichodea and sensilla campaniformia, always present with denticles, are present on the middle labial segment. Three types of sensilla basiconica, two types of sensilla trichodea and two oval flattened sensilla are located on the distal labial segment. The mouthpart morphology and abundance of sensilla located on the labium in C. chinensis are illustrated, along with a brief discussion of their taxonomic and putative functional significance.     

高岭土颗粒涂布对中国梨喀木虱寄主选择、产卵的影响及应用效果

为探索可替代化学药剂防治中国梨喀木虱的措施,开展了高岭土颗粒涂布对其寄主选择、产卵影响及应用效果的研究。分别采用3.5%高岭土水溶液、清水对梨休眠枝和当年生枝叶进行喷雾涂布处理,于24、48、72、96 h监测选择条件下成虫分布情况,于96 h记录选择和非选择条件下各枝条上的卵量。采用高岭土水溶液对梨苗均匀喷雾涂布,通过被害叶率、单叶若虫量、虫龄指数、为害指数对应用效果进行评价。研究结果显示,在选择条件下,经高岭土涂布的枝条上成虫数量明显少于对照,高岭土涂布的枝条上冬型成虫数量在96 h时最低(0.5头/枝),96 h时着卵量(1.83头/枝)显著低于对照(30.83头/枝);高岭土涂布的枝叶上夏型成虫在72 h时最低(1.17头/枝),96 h时着卵量(12.33头/枝)显著低于对照(242.5头/枝);在非选择条件下,冬型和夏型成虫在高岭土涂布的枝条上着卵量(2.83和15.17头/枝)分别显著低于对照(25.17和170.42头/枝);供试植株在接虫45 d后为害最严重,高岭土涂布的植株被害叶率(54.17%)、单叶若虫量(2.29头/叶)、虫龄指数(3.79)、为害指数(0.41)分别显著低于为对照植株(98.61%、11.72头/叶、4.13、0.95)。上述结果表明,高岭土涂布对中国梨喀木虱成虫的寄主选择具有驱避效果,对雌成虫的产卵具有抑制作用;高岭土涂布可限制中国梨喀木虱种群增长,降低危害水平。研究结果为中国梨喀木虱的综合治理提供了现实依据。     

Characterization of microsatellite loci in Cacopsylla melanoneura Förster (Homoptera: Psyllidae)

Cacopsylla melanoneura is one of the main vectors of ‘Candidatus Phytoplasma mali’. We developed and described 10 microsatellite markers. Forty‐five psyllids from one population were analysed. The number of alleles ranged from six to 24 and the observed heterozygosity from 0.42 to 0.84. These markers will be useful tools for population genetic studies on C. melanoneura.     

Anterior alimentary canal of the pear Psylla,Psylla pyricola foerster (Homoptera,Psyllidae)

Scanning and light microscopy investigations of the anterior alimentary canal of the pear psylla, Psylla pyricola Foerster (Homoptera: Psyllidae), revealed the morphology of the labium and stylets, as well as the presence of sensory structures and a valve in the precibarium. The labium consists of three telescoping segments with an internal labial groove, which surrounds and supports the stylet bundle. Also a part of the labial groove is the internal labial clamp. The stylet bundle is comprised of paired styliform mandibles and maxillae, which interlock to form the food and salivary canals. The stylet bundle proximal to the labium forms a large loop within a membranous crumena. When fully retracted the coiled stylets are under tension. Stylet extension generates increasing tension so that when retracted the stylets readily recoil within the crumena. Penetration of leaf tissues by the stylet bundle is dependent on the interaction between stylet muscles, opening and closing of the labial clamp, the barbed stylet tips, and the ventral position of the labium. Proximal to the crumena the paired stylets separate and diverge at the entrance of the precibarium, which is formed by the interlocking of the epi-and hypopharynges. There are 18 sensory structures in the precibarium, as well as a precibarial valve. These structures appear to be homologous to similar structures observed previously in aphids and leafhoppers. The morphology and the location of the precibarial sensilla suggest that, like the precibarial sensory organs of aphids and leafhoppers, they are gustatory and probably mediate acceptance or rejection of plant fluids, thus playing a major role in locating tissues for feeding.     

Descriptions of two new and one little known species of the genus Cacopsylla (Hemiptera: Psyllidae) on Sorbus japonica (Rosaceae) and an observation of their biology

Three species of the genus Cacopsylla were collected from Sorbus japonica in Japan. Two of them, Cacopsylla elegans and Cacopsylla sorbicoccinea, are described as new species based on adults and fifth (final) nymphal instars. The third species, Cacopsylla midoriae, is newly transferred from the genus Psylla. The adult of C. midoriae is redescribed and the nymph is described for the first time. A key to the species feeding on S. japonica (adults and nymphs) is provided. The three species were observed to coexist on the same tree of S. japonica in northern Kyushu. Such coexistence of psylloid congeners is uncommon in Japan.     

New species of Acizzia Heslop-Harrison (Hemiptera: Psyllidae) from Australian mistletoe (Loranthaceae)

Three new species of Acizzia are described from Australian mistletoe (Loranthaceae: Amyema spp.). These are: A. loranthacae sp. n., A. amyemae sp. n. and A. pendulae sp. n. They are characterised by: male proctiger with prominent posterior lobe; antenna 1.9–2.9 times width of head; forewing with costal break and pterostigma, with pattern in A. amyemae and A. pendulae but not in A. loranthacae ; and female proctiger simple in A. amyemae and A. pendulae but with prominent dorso-apical process in A. loranthacae . The Loranthaceae is a new host family record for Acizzia . Given the probable radiation of Acizzia on mimosaceous hosts, its occurrence on eucalypt-inhabiting mistletoe, yet its apparent absence from mistletoes on Acacia, is discussed.     

Dispersion patterns and sampling plans for Diaphorina citri (Hemiptera: Psyllidae) in citrus

The abundance and spatial dispersion of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) were studied in 34 grapefruit (Citrus paradisi Macfad.) and six sweet orange [Citrus sinensis (L.) Osbeck] orchards from March to August 2006 when the pest is more abundant in southern Texas. Although flush shoot infestation levels did not vary with host plant species, densities of D. citri eggs, nymphs, and adults were significantly higher on sweet orange than on grapefruit. D. citri immatures also were found in significantly higher numbers in the southeastern quadrant of trees than other parts of the canopy. The spatial distribution of D. citri nymphs and adults was analyzed using Iowa's patchiness regression and Taylor's power law. Taylor's power law fitted the data better than Iowa's model. Based on both regression models, the field dispersion patterns of D. citri nymphs and adults were aggregated among flush shoots in individual trees as indicated by the regression slopes that were significantly >1. For the average density of each life stage obtained during our surveys, the minimum number of flush shoots per tree needed to estimate D. citri densities varied from eight for eggs to four flush shoots for adults. Projections indicated that a sampling plan consisting of 10 trees and eight flush shoots per tree would provide density estimates of the three developmental stages of D. citri acceptable enough for population studies and management decisions. A presence-absence sampling plan with a fixed precision level was developed and can be used to provide a quick estimation of D. citri populations in citrus orchards.     

Study of COI sequences from endemic New Zealand aphids highlights high mitochondrial DNA diversity in Rhopalosiphina (Hemiptera: Aphididae)

Focussed searches were made across New Zealand between 2013 and 2016, for endemic aphids from the Schizaphis (Rhopalosiphina) genus, which is currently represented by two putative, undescribed species from the endemic host plants Aciphylla and Dracophyllum. Cytochrome c oxidase I (COI) gene sequences (48 in total) from the Schizaphis were analysed together with those from a broader collection of New Zealand endemic aphids that has been assembled since the year 2000. The bulk of the Schizaphis belonged to two clusters corresponding to the host plant genera. Two aphids from central North Island Dracophyllum represented a much diverged lineage without clear affiliations to other New Zealand Schizaphis. Inter-population variation in the New Zealand Schizaphis was high compared with that seen in international studies of Aphidinae and among populations of other endemic New Zealand Aphidina. Within Schizaphis from Dracophyllum, geography played an apparent role in genetic structuring, with populations from Taranaki (North Island) and especially Mt Lyford (South Island) being divergent from those on the South Island main divide. Two distinct lineages of Schizaphis, which co-occurred at some sites, were found on Aciphylla. Our sequence comparisons, including GMYC analyses, indicated up to five New Zealand Schizaphis lineages, and two newly discovered endemic Aphis species from the host plants Clematis and Hebe.