Differential impact of egg predation by Zetzellia mali (Acari: Stigmaeidae) on Metaseiulus occidentalis and Typhlodromus pyri (Acari: Phytoseiidae)

The differential impact of Zetzellia mali on the phytoseiids Metaseiulus occidentalis and Typhlodromus pyri was studied in the laboratory and by analysis of population from orchard plots that contained either phytoseiid, similar numbers of prey mites and high or low densities of Z. mali. Five hypotheses were evaluated to explain why Z. mali had more impact on M. occidentalis in the field than on T. pyri. Given equal opportunity, Z. mali adult females did not consume more M. occidentalis eggs than T. pyri eggs nor did adult females of either phytoseiid inflict greater mortality on Z. mali eggs or larvae through attack or consumption. There was no difference in the within-tree association of Z. mali adult females with eggs of either phytoseiid species nor were there differences in the way prey mites (all life stages) were spatially partitioned between adult female Z. mali as compared with adults and deutonymphs (combined) of either phytoseiid. The foraging area of adult female Z. mali and the oviposition locations of the two phytoseiids from both field and laboratory data were compared using spatial statistics. Metaseiulus occidentalis laid significantly more eggs in the primary foraging area of adult female Z. mali than T. pyri. This was the only factor identified which may explain the greater impact of Z. mali on M. occidentalis. The impact of these interspecific effects on the persistence of predatory mite guilds and biological control are discussed.     

Agrilus mali Matsumara (Coleoptera: Buprestidae), a new invasive pest of wild apple in western China: DNA barcoding and life cycle

Agrilus mali Matsumara (Coleoptera: Buprestidae) is a wood‐boring beetle distributed to eastern China that occasionally injures apple species. However, this wood‐boring beetle is new to the wild apple forests (Malus sieversii) of the Tianshan Mountains (western China) and has caused extensive tree mortality. The development of a biological control program for these wild apple forests is a high priority that requires exploration of the life cycle, DNA barcoding and taxonomic status of A. mali. In this study, to determine the diversity of invasive beetles, a fragment of the mitochondrial cytochrome oxidase gene was analyzed. Based on the results, beetles from Gongliu and Xinyuan counties of Xinjiang were identical but differed from those in the apple nursery of Gongliu by a single‐nucleotide substitution. We summarize the taxonomic status, relationships, and genetic distances of A. mali among other Agrilus species using the Tajima‐Nei model in maximum likelihood phylogeny. Analysis revealed that A. mali was closely related to Agrilus mendax and both belong to the Sinuatiagrulus subgenus. The life cycle of A. mali was investigated based on a monthly regular inspection in the wild apple forests of Tianshan. Similar to congeneric species, hosts are injured by larvae of A. mali feeding on phloem tissue, resulting in serpentine galleries constructed between bark and xylem that prevent nutrient transport and leading to tree mortality. Future studies will focus on plant physiological responses to the invasive beetles and include surveys of natural enemies for a potential classical biological control program.     

Searching behavior and associational response ofZetzellia mali (Acarina: Stigmaeidae)

Experimental tests of the hypothesis that natural selection favors predators that search and select prey species that are more profitable are reported. The predator studies was the stigmaeid mite,Zetzellia mali (Ewing). It was observed thatAculus schlechtendali (Nalepa) andPanonychus ulmi (Koch) were more profitable prey thanTetranychus urticae Koch. However,Z. mali attacks prey independently of their nutritional value. Behavioral observations and experiments documented thatZ. mali does not detect kairomones of the prey species. Therefore, prey location is due only to random encounters. Apparently,Z. mali does not search and select prey that are more profitable. The correlation coefficients ofZ. mali, A. schlechtendali, andP. ulmi were investigated in an apple tree. Results revealed that the association depends on season and type of prey.Zetzellia mali andA. schlechtendali were positively associated in the spring (r=0.3,P<0.05) and fall (r=0.36,P<0.01).Zetzellia mali andP. ulmi were not statistically associated. The term associational response is proposed as an ecological process that quantifies the association of predator and prey. The associational-response data suggest thatZ. mali, in the natural ecosystem, responds more to the density ofA. schlechtendali than to the density ofP. ulmi.     

Life history and mortality factors of Agrilus mali Matsumura (Coleoptera: Buprestidae) in wild apples in Northwestern China

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Geographic Variation of Esterase Isozymes in Populations of Alternaria mali on Apple Leaves

The 500 monoconidial isolates of Alternaria mali occurring in different locations, Suweon, Cheongju, Kochang, Daegu and Jinju, Korea, in 1983 were used to examine geographic variation of esterase isozymes. The electrophoretic patterns of esterases were qualitatively and quantitatively different between isolates. The 14 different bands were detected on the basis of the decreasing electrophoretical mobility, although all bands were not present in any of the isolates. A comparison of the frequency of esterase isozymes at different bands showed marked variations among the geographic locations. The geographic distance between A. mali populations did not correlate strongly with divergence in esterase isozymes, whereas A. mali populations within a geographic feature were more closely related than populations separated by a mountain range.     

Phoretic dispersal ofBrennandania lambi (Kcrzal) (Acari: Tarsonemida: Pygmephoridae) by mushroom flies (Diptera: Sciaridae and Phoridae) in New South Wales,Australia

The phoridMegaselia halterata (Wood) was found to carry significantly moreBrennandania lambi (Kerzal) than the sciaridLycoriella mali (Fitch) in samples collected from six mushroom farms.B. lambi was attached to both the phorids and mainly between the procoxae and mesosternum (57&) and between metacoxae and the first abdominal sternite (36%).B. lambi detached fromM. halterata only in the presence ofAgaricus mycelium. Evidence for dispersal ofB. lambi byL. mali andM. halterata is presented and the role of phoresy in the dispersal ofB. lambi is discussed.     

Assessment of real-time PCR for quantification of Legionella spp. in spa water

Aim: To identify media and environmental conditions suitable for rapid mycelial growth and sporulation of Diplocarpon mali. Methods and Results: Liquid shake cultures were used to evaluate effects of media and environmental conditions on mycelial growth and conidial production of D. mali. Carrot sucrose broth (CSB), potato and carrot dextrose broth (PCDB) and potato and carrot sucrose broth (PCSB) were most favourable for rapid mycelial growth. PCDB, PCSB, PCB (potato and carrot broth) and carrot dextrose broth (CDB) were favourable for conidial production. All carbon sources tested and peptone favoured for mycelial growth. Carbon and nitrogen sources tested did not significantly stimulate conidial production. The optimum temperature for mycelial growth and conidial production was 25°C. No mycelial growth occurred at 5 or 30°C, but D. mali survived at these temperatures. Active mycelial growth occurred at pH 5–7, and pH 5–8 was favourable for sporulation. Conclusions: PCDB and PCSB incubated at 25°C for 14 day are recommended for mycelial growth and conidial production of D. mali. Significance and Impact of the Study: The information generated in this study will facilitate mycological and pathological research on D. mali and Marssonina leaf blotch of apple caused by D. mali.     

Effective cellulose production by a coculture of Gluconacetobacter xylinus and Lactobacillus mali

A microbial colony that contained a marked amount of cellulose was isolated from vineyard soil. The colony was formed by the associated growth of two bacterial strains: a cellulose-producing acetic acid bacterium (st-60-12) and a lactic acid bacterium (st-20). The 16S rDNA-based taxonomy indicated that st-60-12 belonged to Gluconacetobacter xylinus and st-20 was closely related to Lactobacillus mali. Cocultivation of the two organisms in corn steep liquor/sucrose liquid medium resulted in a threefold higher cellulose yield when compared to the st-60-12 monoculture. A similar enhancement was observed in a coculture with various L. mali strains but not with other Lactobacillus spp. The enhancement of cellulose production was most remarkable when sucrose was supplied as the substrate. L. mali mutants for exocellular polysaccharide (EPS) production were defective in promoting cellulose production, but the addition of EPS to the monoculture of st-60-12 did not affect cellulose productivity. Scanning electron microscopic observation of the coculture revealed frequent association between the st-60-12 and L. mali cells. These results indicate that cell–cell interaction assisted by the EPS-producing L. mali promotes cellulose production in st-60-12.The nucleotide sequences of 16S rDNA that are reported in this paper were submitted to GenBank/EMBL/DDBJ under the accession numbers AB016864 (st-20) and AB016865 (st-60-12).     

Variability in Sensitivity to Polyoxin B of Isolates of Alternaria mali and Decreased Fitness of Polyoxin resistant Isolates

The 540 monoconidial isolates of Altemaria mali were obtained in 1983 from apple orchards at Seoul, Suweon, Cheongju, Kochang, Daegu, and Jinju in Korea. The sensitivity of A. mali to polyoxin B greatly varied among isolates and locations. Most of isolates were sensitive to polyoxin B, but 11 isolates showed a high level of resistance, particularly from Kochang and Daegu where polyoxin B had been applied frequently. Conidial germination and mycelial growth of resistant isolates were not inhibited at higher concentrations of polyoxin B compared to the sensitive ones. The polyoxin-resistant isolates were not resistant to the fungicides iprodione and polydong and showed a reduced activity for conidial formation and mycelial growth. The isolates of A. mali with resistance to polyoxin B appear to have decreased fitness in nature.     

Development of Apheliuus mali an endoparasitoid of woolly apple aphid, Eriosoma lanigerum at different temperatures

Developmental times for both sexes of Aphelinus mali (Haldeman) (Hymenoptera: Aphelinidae) an endoparasitoid of woolly apple aphid, Eriosoma lanigerum (Hausmann) (Hemiptera: Pemphigidae) were studied at 13, 15, 18, 20, 25 and 30°C and compared with those of E. lanigerum. Mean developmental times ranged from 11.7–53.3 days for males, 11.8–55.4 days for females and 11.8–54.4 days for both sexes combined and were significantly inversely related to temperature. A good linear model fit (r²>0.99) between developmental rate and temperature in the range 13–30°C was observed. Results indicate significant differences between developmental times of males and females at 13, 18, 20, and 25°C but no differences at 15 and 30°C. The notional developmental threshold was 8.3°C for both males and females. Compared with its host, A. mali has a higher lower developmental threshold. An average of 252.8, 256.7 and 254.8 degree-days (DD) above the lower threshold were required by A. mali to complete development from time of oviposition to adult emergence for males, females and both sexes combined, respectively. Field experimentation also indicated that the developmental time of A. mali lags significantly behind that of its aphid host throughout the year. These findings are discussed in relation to its status as a biological control agent for E. lanigerum.     

新疆野苹果AP2/ERF转录因子家族鉴定与响应腐烂病的表达分析北大核心CSCD

AP2/ERF转录因子家族参与了植物生长发育、抵抗胁迫以及植物激素响应等诸多生物过程,是植物中最重要的转录因子家族之一。该研究基于腐烂病菌侵染后的新疆野苹果(Malus sieversii)全长转录组,使用AP2保守结构域的隐马可夫模型PF00847,鉴定新疆野苹果的AP2/ERF家族成员。利用MEGA6、NCBI CDD-batch、MEME、EXPASY、BUSCA对MsAP2/ERF家族成员进行鉴定、分类和结构分析、理化性质和亚细胞定位分析。通过RNA-seq数据和qRT-PCR实验对差异表达的MsAP2/ERFs基因的表达水平进行分析和验证,旨在鉴定新疆野苹果中潜在具有腐烂病抗性的AP2/ERF家族基因资源。结果显示:(1)在新疆野苹果中共鉴定获得106个AP2/ERF基因,涵盖全部AP2(17个)、ERF(57个)、DREB(25个)、RAV(5个)和Soloist(2个)5个亚家族。(2)进一步的细化分类发现MsERF亚家族包括B1-B6 6个组,而MsDREB亚家族中只有A2、A4、A5、A6共4个组,缺少A1和A3组的基因成员。(3)RNA-seq表达模式分析结果表明,29个MsAP2/ERF基因在腐烂病感染过程中差异表达,其中MsERF亚家族中差异表达基因数量最多(19个)。(4)12个MsAP2/ERF代表基因的qRT-PCR结果表明:8个ERF亚家族基因均受腐烂病病菌诱导显著上调表达,其中B4类ERF成员基因(MsERF40)在腐烂病病菌侵染后5 d表达量上调表达倍数最高;4个MsDREB基因中,3个受到腐烂病病原菌诱导显著上调,1个下调表达;此外,含有TIR保守结构域的MsERF05在腐烂病病菌侵染1 d后上调表达69倍,表明ERF亚家族的MsERF40和MsERF05在新疆野苹果抗腐烂病过程中发挥重要作用。该研究结果为新疆野苹果AP2/ERF基因响应腐烂病的功能和机理研究奠定了基础。     

Hce2 domain-containing effectors contribute to the full virulence of Valsa mali in a redundant manner

Valsa mali is the causal agent of apple Valsa canker, a destructive disease in East Asia. Effector proteins play important roles in the virulence of phytopathogenic fungi, and we identified five Hce2 domain-containing effectors (VmHEP1, VmHEP2, VmHEP3, VmHEP4 and VmHEP5) from the V. mali genome. Amongst these, VmHEP1 and VmHEP2 were found to be up-regulated during the early infection stage and VmHEP1 was also identified as a cell death inducer through its transient expression in Nicotiana benthamiana. Although the deletion of each single VmHEP gene did not lead to a reduction in virulence, the double-deletion of VmHEP1 and VmHEP2 notably attenuated V. mali virulence in both apple twigs and leaves. An evolutionary analysis revealed that VmHEP1 and VmHEP2 are two paralogues, under purifying selection. VmHEP1 and VmHEP2 are located next to each other on chromosome 11 as tandem genes with only a 604 bp physical distance. Interestingly, the deletion of VmHEP1 promoted the expression of VmHEP2 and, vice versa, the deletion of VmHEP2 promoted the expression of VmHEP1. The present results provide insights into the functions of Hce2 domain-containing effectors acting as virulence factors of V. mali, and provide a new perspective regarding the contribution of tandem genes to the virulence of phytopathogenic fungi.     

Firewood collected after a catastrophic wind event: the bark beetle (Scolytinae) and woodborer (Buprestidae,Cerambycidae) community present over a 3‐year period

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Addition and reexamination of Japanese species belonging to the genus <Emphasis Type="Italic">Cercospora</Emphasis> and allied genera. VII. Newly recorded species from Japan (2)

In the seventh report of the present series, four species of the genus Pseudocercospora were added to the Japanese mycoflora. They are Pseudocercospora catappae, P. mali, P. polysciatis-pinnatae, and P. puderi.     

苹果树腐烂病菌非核糖体多肽合成酶基因VmNRPS12的功能

【目的】非核糖体多肽合成酶(NRPS)在植物病原真菌与其寄主互作过程中发挥着重要作用,明确Vm NRPS12基因在苹果树腐烂病菌致病过程中的功能,将为今后深入研究苹果树腐烂病菌NRPS作用机制提供理论依据。【方法】基于苹果树腐烂病菌全基因组数据,得到VmNRPS12基因。运用qRT-PCR技术分析VmNRPS12在侵染初期的表达水平,利用Double-joint PCR和PEG介导的原生质体转化获得该基因抗潮霉素的突变体,对突变体进行PCR检测及Southern blot验证得到敲除突变体,进一步通过重新导入该基因全长片段获得互补突变体,最后对野生型、敲除突变体和互补突变体进行菌落、产孢及致病力观察,对检测数据用SPSS软件进行差异显著性分析。【结果】定量分析显示该基因在侵染初期显著上调表达,且接种48 h后的表达量是对照的138.6倍。该基因的敲除突变体在营养生长及产孢方面与野生型菌株03-8相比无显著性差异,但致病力与野生型菌株03-8相比显著减弱,且互补突变体致病力近似恢复至野生型水平。【结论】VmNRPS12基因与苹果树腐烂病菌致病性相关。     

Predation on Mindarus abietinus infestingbalsam fir grown as Christmas trees: the impact ofcoccinellid larval predation with emphasis on Anatis mali

The impact of natural coccinellid larvalpredation on the balsam twig aphid was evaluated bysystematically removing coccinellid egg masses in a6–8 year-old balsam fir (Abies balsamea)Christmas tree plantation in southwesternQuebec. Among coccinellid species hunting on firfoliage during development of Mindarus abietinusfundatrices in May, the indigenous Anatis mali was by far the most abundant and themain one to oviposit on trees. Comparison of trees onwhich coccinellid larval predation was excluded withcontrol trees showed that A. mali had a markedimpact both during and after the phase of rapid M. abietinus population growth that followedfundatrix maturation. On trees where coccinellidlarvae were allowed, aphid colonies became inactive(i.e. no live aphids in the colony) about two weeksearlier than on controls. A strong dampening effect onaphid density was also observed in those colonies thatremained active until the end of the aphid life cycle.Predation on aphid colonies reduced sexualsproduction, as the density of M. abietinusoverwintering eggs per shoot subsequently was reducedby 32%. Predation by coccinellid larvae occurred toolate to prevent needle damage to current year shoots,which affects the aesthetic value of Christmas trees.However, current year shoots measured in the mid-crownof trees late in the season were 19% longer on treeswhere aphid predation by coccinellid larvae wasallowed, compared with trees where they were excluded.Rearing all larval stages of A. mali on 4thinstar and adult sexuparae of M. abietinusindicated an average consumption of 269 aphids tocomplete larval development and pupate, which wasequivalent to at least seven colonies of M.abietinus at maximum aphid density at theexperimental site. Anatis mali is an importantnatural control factor of balsam twig aphid inChristmas tree plantations, hence its activity shouldbe protected and possibly stimulated by favourablepest management practices.     

Two new species of Stemphylium from Northwest China

During surveys of hyphomycetes from the Northwest of China, two undescribed taxa of Stemphylium were discovered from diseased leaves of Malus sieversii and Pyrus sinkiangensis in Sinkiang Province, which are described as S. mali and S. pyrinum. The two species differ from similar species morphologically.     

Feeding preference of Zetzellia mali: does absolute or relative abundance of prey matter more?

The importance of the acarine predator, Zetzellia mali, in the control of phytophagous mites in apple orchards is not well understood. Zetzellia mali tends to prefer the eriophyid, Aculus schlechtendali, over the economically more significant tetranychid, Panonychus ulmi, but quite a wide range of preference values have been reported in the literature. In sets of laboratory choice trials, we determined that prey preference of this predator varies with the relative but not absolute density of its prey. We attempt to explain these results in terms of behavioural mechanisms and discuss the potential implications of our results for the effectiveness of Z. mali in the biological control of phytophagous mites in apple orchards.