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.     

‘嘎拉’苹果及其杂交后代对炭疽叶枯病的抗性机制分析

为解析苹果对炭疽叶枯病的抗性机制,该研究以‘嘎拉’、‘藤牧1号’苹果及其杂交后代等87个苹果品种(系)为试验材料,进行田间调查及人工接种病菌并检测不同品种(系)中病菌生物量,利用SSR分子标记进行基因型鉴定,分析各品种(系)对炭疽叶枯病的抗性差异,利用荧光定量PCR及酶活检测比较分析水杨酸相关基因、抗性酶基因及酶活性水平差异。结果表明：(1)87个苹果品种(系)对苹果炭疽叶枯病的抗性差异明显,‘嘎拉’、‘2 5’、‘19 19’等品种(系)叶片发病严重,表现出对炭疽叶枯病的高感性,‘藤牧1号’、‘40 9’及‘16 16’等品种(系)叶片无病斑或病斑极少,炭疽叶枯病菌含量显著低于感病性品种(系),其抗性显著。(2)SSR标记S0405127和S0304673的基因型鉴定结果与田间表型调查结果相比,准确率分别为93.10%和91.95%,与人工接种结果相比,准确率分别为91.95%和95.40%。(3)SA相关基因的表达模式结果表明,接种炭疽叶枯病菌4 d后,‘藤牧1号’、‘40 9’及‘16 16’等抗性品种(系)中SA合成相关基因MdEDS1、MdPAD4和MdPAL被强烈诱导表达;同时,SA信号转导相关基因MdNPR1、MdPR1、MdPR5的表达显著高于‘嘎拉’等感病品种(系)。(4)接种炭疽叶枯病菌4 d后,‘藤牧1号’、‘40 9’及‘16 16’等抗性品种(系)的MdSOD、MdPOD酶基因表达水平及酶活性显著高于‘嘎拉’、‘2 5’、‘19 19’等感病品种(系)。研究认为,‘藤牧1号’、‘40 9’及‘16 16’等品种(系)通过调控水杨酸合成和信号转导通路及氧化还原相关反应等提高对炭疽叶枯病的抗性,为挖掘抗性基因以及利用优良种质选育抗病品种奠定了基础。     

Potential host fruits for Drosophila suzukii: olfactory and oviposition preferences and suitability for development

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), the spotted wing drosophila, is a pest endemic to Southeast Asia that invaded the Americas and Europe in 2008. In contrast to most of its congeners, D. suzukii possesses a serrated ovipositor that allows it to lay eggs in unwounded commercial fruits, resulting in severe revenue losses for the industry. The purpose of this study was to determine the susceptibility of known host fruits, including cherry, strawberry, blueberry, and grape, and potential host fruits, such as banana and apple, to attack by D. suzukii. Based on the responses to volatile cues offered in a six‐choice olfactometer, the preference of female D. suzukii was ranked in the following order: strawberry = cherry > banana = apple = blueberry = grape, but in no‐choice and choice oviposition tests, the preferences were ranked as follows: cherry > strawberry = blueberry > grape = banana > apple. Furthermore, we reconfirmed that D. suzukii mainly targets rotten fruit for feeding and ripe fruit for oviposition, and females preferred fruits with intensive mechanical damage. Based on developmental parameters, apple was the least suitable host. This study has implications for the control of D. suzukii, especially in mixed fruit orchards, by providing a promising avenue for exploiting behaviour‐based control tools and emphasizing the importance of phenology in host fruit susceptibility.     

Natural variation in cytokinin maintenance improves salt tolerance in apple rootstocks

Plants experiencing salt‐induced stress often reduce cytokinin levels during the early phases of stress‐response. Interestingly, we found that the cytokinin content in the apple rootstock “robusta” was maintained at a high level under salt stress. Through screening genes involved in cytokinin biosynthesis and catabolism, we found that the high expression levels of IPT5b in robusta roots were involved in maintaining the high cytokinin content. We identified a 42 bp deletion in the promoter region of IPT5b, which elevated IPT5b expression levels, and this deletion was linked to salt tolerance in robusta×M.9 segregating population. The 42 bp deletion resulted in the deletion of a Proline Response Element (ProRE), and our results suggest that ProRE negatively regulates IPT5b expression in response to proline. Under salt stress, the robusta cultivar maintains high cytokinin levels as IPT5b expression cannot be inhibited by proline due to the deletion of ProRE, leading to improve salt tolerance.     

Spring colonisation of orchards by Anthonomus pomorum from adjacent forest borders

The early-season dispersal of the overwintered apple blossom weevil, Anthonomus pomorum (L.) (Coleoptera: Curculionidae), is a crucial stage in the colonisation of dwarf apple orchards adjacent to forests. We have conducted release-recapture studies with 1700 to 4000 marked weevils at two orchard sites in Switzerland over 2 years to characterise the spatial and temporal pattern of the dispersal process. The dispersal and colonisation of orchards in spring by overwintered weevils is dependent upon the prevailing temperature.An orientated dispersal from the forest border to the centre of the orchard was observed consistently, irrespective of the angle of the apple tree rows with respect to the forest border or of climatic conditions. The average dispersal distance of the weevils was 19 m. Approximately one third of the weevil population remained on the first tree encountered, the remainder of the population moved over short distances mainly along the tree rows. This dispersal pattern led to a strong edge effect with higher numbers of weevils occurring at the edges adjoining the forests as compared to the centre of orchards. The relevance of these findings to population dynamics and management of the pest is discussed.     

It''s about time: the evidence for host plant-mediated selection in the apple maggot fly, Rhagoletis pomonella, and its implications for fitness trade-offs in phytophagous insects

The apple maggot fly, Rhagoletis pomonella, Walsh (Diptera: Tephritidae), provides a unique opportunity to address the issue of host-related fitness trade-offs for phytophagous insects. Rhagoletis pomonella has been controversial since the 1860's when Benjamin Walsh cited the fly's shift from hawthorn (Crataegus spp.) to apple (Malus pumila) as an example of an incipient sympatric speciation event. Allozyme and mark-release-recapture studies have subsequently confirmed the status of apple and hawthorn flies as partially reproductively isolated and genetically differentiated host races, the hypothesized initial stage in sympatric divergence. Here, we review the ecological and genetic evidence for host-plant mediated selection in R. pomonella. We reach the following three major conclusions: First, although developmental timing is not everything, it is a good deal of the story. Differences in the fruiting phenologies of apple and hawthorn trees exert different selection pressures on the diapause and eclosion time characteristics of the host races. In particular, the 3-week earlier mean fruiting phenology of apples in eastern North America appears to select for a slower rate of metabolism or deeper pupal diapause in apple than hawthorn flies. Second, host-related fitness trade-offs for R. pomonella may not be due to disruptive selection affecting any one specific life-history stage. Rather, it is the sum total of directional selection pressures acting across different life-stages that generates divergent selection on apple and hawthorn flies. For example, selection favors the alleles Me 100, Acon-2 95 and Mpi 37 (or linked genes) in the larval stage in both host races. However, these same alleles are disfavored in the pupal stage to follow, where they correlate with early adult eclosion, and by inference premature diapause termination. Because apple trees fruit an average of 3 weeks earlier than hawthorn trees, this counter-balancing selection is stronger on apple-fly pupae. The net result is that the balance of selective forces is different between apple and hawthorn flies, helping to maintain the genetic integrity of the host races in sympatry in the face of gene flow. Finally, natural R. pomonella populations harbor a good deal of genetic variation for development-related traits. This variation allows fly populations to rapidly respond to temporal vagaries in local environmental conditions across years, as well as to broad-scale geographic differences that exist across the range of the species. Perhaps most importantly, this variation gives R. pomonella the flexibility to explore and adapt to novel plants. Taken together, our results underscore how difficult it can be to document host plant-related fitness trade-offs for phytophagous insects due to the need to consider details of the entire life-cycle of a phytophagous insect. Our findings also show how reproductive isolation can arise as a by-product of host-associated adaptation in insects, a central theme for models of sympatric speciation via host shifts.     

AFLP-derived SCARs facilitate construction of a 1.1 Mb sequence-ready map of a region that spans the Vf locus in the apple genome

The availability of high-density anchored markers is a prerequisite for reliable construction of a deep coverage BAC contig, which leads to creation of a sequence-ready map in the target chromosomal region. Unfortunately, such markers are not available for most plant species, including woody perennial plants. Here, we report on an efficient approach to build a megabase-size sequence-ready map in the apple genome for the Vf region containing apple scab resistance gene(s) by targeting AFLP-derived SCAR markers to this specific genomic region. A total of 11 AFLP-derived SCAR markers, previously tagged to the Vf locus, along with three other Vf-linked SCAR markers have been used to screen two apple genome BAC libraries. A single BAC contig which spans the Vf region at a physical distance of approximately 1,100 kb has been constructed by assembling the recovered BAC clones, followed by closure of inter-contig gaps. The contig is 4 ×deep, and provides a minimal tiling path of 16 contiguous and overlapping BAC clones, thus generating a sequence-ready map. Within the Vf region, duplication events have occurred frequently, and the Vf locus is restricted to the ca. 290 kb region covered by a minimum of three overlapping BAC clones.