Development of multiplex RT-PCR assay for simultaneous detection of four viruses infecting apple (Malus domestica)

The major viruses infecting apple cultivars throughout the world including India are apple mosaic virus (ApMV), apple stem pitting virus (ASPV), apple stem grooving virus (ASGV), apple chlorotic leaf spot virus (ACLSV), and recently, a new virus, apple necrotic mosaic virus (ApNMV), was reported from mosaic-infected apple cultivars in India. The aim of this study was to detect the ApNMV virus along with the other three viruses (ApMV, ASPV and ASGV) simultaneously by multiplex RT-PCR. Four primer-pair-produced amplicons of 670, 550, 350 and 210 bp corresponding to ApNMV, ApMV, ASPV and ASGV, respectively, were found to be specific for these viruses when tested individually. The annealing temperature (55°C), primer concentration (0·8 µl) and other components of the master mix were standardized for the development of one-step m-RT-PCR assay. The m-RT-PCR protocol developed was further validated with 30 samples from seven symptomatic or asymptomatic apple cultivars, which revealed the presence of more than one virus in these cultivars. Most of the viruses were found to be present either alone or in mixed infection; however, ASPV was more common in tested cultivars. An easy, cost-effective and rapid multiplex RT-RCR protocol was developed to detect the four viruses, which infect apple plants either in individually or together in the field. This assay will help in the surveying and indexing of apple germplasm and the distribution of all four viruses in the apple growing regions of India.     

Plant volatiles mediate attraction to host and non-host plant in apple fruit moth, Argyresthia conjugella

Plant volatiles mediate host finding in insect herbivores and lead to host fidelity and habitat‐specific mating, generating premating reproductive isolation and facilitating sympatric divergence. The apple fruit moth, Argyresthia conjugella Zeller (Lepidoptera: Argyresthiidae), is a particularly suitable species to study the cues and behavioural mechanisms leading to colonization of a new host: it recurrently oviposits on the non‐host plant, apple Malus domestica Borkh. (Rosaceae), where the larvae cannot complete their development. The larval host of the apple fruit moth (Lepidoptera, Argyresthiidae), is rowan Sorbus aucuparia L. (Rosaceae). Fruit setting in rowan, however, fluctuates strongly over large areas in Scandinavia. Every 2–4 years, when too few rowanberries are available for egg laying in forests, apple fruit moth females oviposit instead on apple in nearby orchards, but not on other fruits, such as pear or plum. This poses the question of which cues mediate attraction to rowan and apple, and how apple fruit moth discriminates rowan from apple. Chemical analysis and antennal recordings showed that 11 out of 15 rowan volatiles eliciting an antennal response in A. conjugella females co‐occur in rowan and apple headspace, in a different proportion. In the field, A. conjugella was attracted to several of these plant volatiles, especially to 2‐phenyl ethanol, methyl salicylate, and decanal. Addition of anethole to 2‐phenyl ethanol had a strong synergistic effect, the 1 : 1 blend is a powerful attractant for A. conjugella males and females. These results confirm that volatiles common to both plants may account for a host switch in A. conjugella from rowan to apple. Some of the most attractive compounds, including 2‐phenyl ethanol, anethole, and decanal, which have been found in several apple cultivars, were not present in the headspace of the apple cultivar, Aroma, which is also susceptible to attack by A. conjugella. This supports the idea that the odour signal from apple is suboptimal for attraction of A. conjugella, but is nonetheless sufficient for attraction, during times when rowan is not available for egg laying.     

Differential susceptibility and suitability of domestic and wild apple species for a florivorous weevil and its parasitoids

Crop plant domestication can change plant resistance to herbivores leading to differences in pest pressure experienced by crop plants and their wild relatives. To compare resistance to herbivores between domesticated and wild fruit trees, we quantified direct resistance and indirect resistance to a pest insect, the florivorous apple blossom weevil Anthonomus pomorum (Coleoptera: Curculionidae), for the cultivated apple Malus domestica and two wild apple species, the European crab apple M. sylvestris and the exotic M. kirghisorum. We measured weevil infestation and performance (weight, sex ratio), and weevil parasitism by parasitoid wasps for different cultivars of M. domestica and for the two wild apple species. To explain weevil and parasitoid responses to different apple species, we quantified tree characteristics including nitrogen content, size of flower buds, bark roughness, tree size, tree phenology and tree position. We found significant differences in susceptibility to weevil infestation between apple species, with lowest infestation (highest apple resistance) in M. domestica and highest infestation in M. kirghisorum. The suitability of apple species also varied significantly: weevils emerging from M. sylvestris were significantly lighter than those from M. kirghisorum. Parasitism of A. pomorum by different parasitoid species was significantly higher in M. sylvestris than in M. domestica. Infestation, weevil weight and parasitism were positively related to tree characteristics: infestation to bud nitrogen content and bark roughness, weevil size to nitrogen content and bud size, and parasitism to tree height and bud density. Our study revealed marked differences between apple species in susceptibility and suitability for the pest herbivore, but also for antagonistic parasitoids. Whereas direct resistance appeared to be higher in cultivated apple, indirect resistance via parasitoids was apparently higher in wild apple trees. Our findings suggest that wild and cultivated apple trees possess different resistance traits that may be combined to optimize resistance in commercial apple cultivars.     

Functional allelic diversity of the apple alcohol acyl-transferase gene <Emphasis Type="Italic">MdAAT1</Emphasis> associated with fruit ester volatile contents in apple cultivars

Flavour is an important key factor of apple (Malus × domestica Borkh.) fruit quality, and its improvement is an important but complex breeding goal. Acetate esters are quantitatively the most important volatile compounds in apple fruit, and only a few of them dominate the typical aroma of a cultivar. Alcohol acyl-transferase (AAT) is a key enzyme involved in the last step of ester biosynthesis. The aim of this study was to target single nucleotide polymorphisms (SNPs) in an AAT candidate gene genetically associated with ester quantitative trait loci (QTL), to enable functional marker development for marker-assisted apple breeding programs. The AAT gene inventory of apple was characterized by in-silico mining of the assembled Golden Delicious genome, and 17 putative AAT genes in total were defined. MdAAT1 located on chromosome 2 was selected as the main candidate gene associated with QTL for different acetate esters, and its allelic diversity was assessed by direct amplicon sequencing in a collection of 102 apple cultivars characterized for ester volatile profiles. Sequencing a 468 bp nucleotide sequence of the MdAAT1 coding region resulted in the detection of four SNPs. In total, 18 different SNP haplotypes/heterozygous patterns were generated from the four SNPs identified within the apple collection. Association analyses resulted in highly significant associations of both individual SNPs and distinct haplotypes with the content of four acetate esters, including hexyl acetate, butyl acetate and 2-methyl-butyl acetate. About a third (31) of the 102 apple cultivars possessed the specific MdAAT1 haplotype H1 (C-A-C-A) and were characterized by strongly decreased ester concentrations. The contrasting H8 haplotype (T-G-T-G) was found in 28 varieties but was associated with normal to elevated ester concentrations. The observed association suggests a putative causal functional relationship between MdAAT1 and production of key apple esters.     

The O‐methyltransferase gene MdoOMT1 is required for biosynthesis of methylated phenylpropenes in ripe apple fruit

Phenylpropenes, such as eugenol and trans‐anethole, are important aromatic compounds that determine flavour and aroma in many herbs and spices. Some apple varieties produce fruit with a highly desirable spicy/aromatic flavour that has been attributed to the production of estragole, a methylated phenylpropene. To elucidate the molecular basis for estragole production and its contribution to ripe apple flavour and aroma we characterised a segregating population from a Royal Gala (RG, estragole producer) × Granny Smith (GS, non‐producer) apple cross. Two quantitative trait loci (QTLs; accounting for 9.2 and 24.8% of the variation) on linkage group (LG) 1 and LG2 were identified that co‐located with seven candidate genes for phenylpropene O‐methyltransferases (MdoOMT1–7). Of these genes, only expression of MdoOMT1 on LG1 increased strongly with ethylene and could be correlated with increasing estragole production in ripening RG fruit. Transient over‐expression in tobacco showed that MdoOMT1 utilised a range of phenylpropene substrates and catalysed the conversion of chavicol to estragole. Royal Gala carried two alleles (MdoOMT1a, MdoOMT1b) whilst GS appeared to be homozygous for MdoOMT1b. MdoOMT1a showed a higher affinity and catalytic efficiency towards chavicol than MdoOMT1b, which could account for the phenotypic variation at the LG1 QTL. Multiple transgenic RG lines with reduced MdoOMT1 expression produced lower levels of methylated phenylpropenes, including estragole and methyleugenol. Differences in fruit aroma could be perceived in these fruit, compared with controls, by sensory analysis. Together these results indicate that MdoOMT1 is required for the production of methylated phenylpropenes in apple and that phenylpropenes including estragole may contribute to ripe apple fruit aroma.     

Alcohol acyl transferase 1 links two distinct volatile pathways that produce esters and phenylpropenes in apple fruit

Fruit accumulate a diverse set of volatiles including esters and phenylpropenes. Volatile esters are synthesised via fatty acid degradation or from amino acid precursors, with the final step being catalysed by alcohol acyl transferases (AATs). Phenylpropenes are produced as a side branch of the general phenylpropanoid pathway. Major quantitative trait loci (QTLs) on apple (Malus × domestica) linkage group (LG)2 for production of the phenylpropene estragole and volatile esters (including 2‐methylbutyl acetate and hexyl acetate) both co‐located with the MdAAT1 gene. MdAAT1 has previously been shown to be required for volatile ester production in apple (Plant J., 2014, https://doi.org/10.1111/tpj.12518 ), and here we show it is also required to produce p‐hydroxycinnamyl acetates that serve as substrates for a bifunctional chavicol/eugenol synthase (MdoPhR5) in ripe apple fruit. Fruit from transgenic ‘Royal Gala’ MdAAT1 knockdown lines produced significantly reduced phenylpropene levels, whilst manipulation of the phenylpropanoid pathway using MdCHS (chalcone synthase) knockout and MdMYB10 over‐expression lines increased phenylpropene production. Transient expression of MdAAT1, MdoPhR5 and MdoOMT1 (O‐methyltransferase) genes reconstituted the apple pathway to estragole production in tobacco. AATs from ripe strawberry (SAAT1) and tomato (SlAAT1) fruit can also utilise p‐coumaryl and coniferyl alcohols, indicating that ripening‐related AATs are likely to link volatile ester and phenylpropene production in many different fruit.     

The ecophysiology of apple snails in rice: implications for crop management and policy

This review relates the ecology and physiology of apple snails (Ampullariidae) to their impact on rice‐production systems. Two species in particular, Pomacea canaliculata and Pomacea maculata, have been introduced to several rice‐growing regions. Flooded rice systems represent a high‐quality habitat for these apple snails because of similarities in the environmental conditions (water temperature, salinity, pH, water flow velocity) necessary for both rice production and for snail survival and development. Furthermore, amphibious respiration, a capacity to aestivate during dry periods, as well as cold acclimation and tolerance (particularly in P. canaliculata), increases the resilience of apple snails to rice farming practices, including agrochemical applications, intermittent drainage and crop rotations – under a wide range of climatic conditions. Risks to regional rice production depend on four principal factors: these are (a) regional climate, (b) regional rice‐production systems, (c) prevailing production practices, and (4) the presence/absence of invasive, non‐native apple snails. Based on these criteria, lowland irrigated rice in tropical and subtropical regions that is wet‐direct seeded is most vulnerable to damage from both native and non‐native apple snails (albeit with greater losses from non‐native snails because of normally higher densities). Adequate quarantine regulations, particularly in vulnerable tropical regions that are adjacent to centres of recent outbreaks (e.g. India and Bangladesh adjacent to Myanmar, Peru and Colombia adjacent to Ecuador) and attention to the preparedness of farming systems could reduce potential impacts as these highly invasive snails continue to spread. The urgent development of labour‐saving crop‐establishment methods that reduce dependencies on chemical molluscicides is necessary to achieve sustainable rice production in regions at risk from non‐native apple snails.     

THE CHROMOSOMES OF THE SEMPERVIVOIDEAE (CRASSULACEAE)

Uhl , Charles H. (Cornell U., Ithaca, New York.) Chromosomes of the Sempervivoideae (Crassulaceae). Amer. Jour. Bot. 48(2): 114–123. Illus. 1961.—Chromosome numbers are reported for 207 collections representing 68 of the ca. 95 species in this subfamily. Basic numbers are 16, 17, 18, and 19 in Sempervivum, Section Sempervivum (10 species, with many tetraploids and one hexaploid); 19 in Sempervivum, Section Jovisbarba (5 species, all diploid); 15 in Aichryson (9 species, including 1 aneuploid, 1 tetraploid, and 1 aneutetraploid); and strictly 18 in Aeonium (31 species, including 4 wholly and 1 partly tetraploid), Greenovia (3 species, 1 partly tetraploid), and Monanthes (10 species, including 2 wholly and 1 partly tetraploid). The cytological evidence appears decisive in ranking several species of disputed generic position definitely with Aichryson rather than with Aeonium. Possible relationships between various Canarian genera and certain North African species often classed in Sedum are discussed briefly in the light of the scanty morphological and cytological evidence. It is suggested that both these groups may be descended from the same ancestors that were widespread in North Africa before the deserts developed.     

SSRs isolated from apple can identify polymorphism and genetic diversity in pear

Apple simple sequence repeats (SSRs) were intergenerically applied to the characterization of 36 pear accessions, including 19 Japanese pears (Pyrus pyrifolia), 7 Chinese pears (P. bretschneideri, P. ussuriensis), 5 European pears (P. communis), 3 wild relatives (P. calleryana), and 2 hybrids between P. pyrifolia and P. communis. All of the tested SSR primers derived from apple produced discrete amplified fragments in all pear accessions. Nucleotide repeats were detected in the amplified bands by both Southern blot and sequencing analysis, and nucleotide sequences of pear were compared with those of apple. The differences in fragment size among pear or between pear and apple were, in many cases, due to the differences in repeat number. Interestingly, the DNA sequence of flanking regions in apple was highly conserved in pear. Hybrids from P. pyrifolia×P. communis showed one fragment inherited from each parent in all scorable cases, which suggested that each primer pair amplified fragments originating from the same locus. A total of 79 alleles were detected from seven SSR loci in pear, and all pear varieties except for the mutants could be differentiated. In conclusion, SSRs isolated from apple are highly conserved in pear and could be utilized as DNA markers in the latter genus. Received: 17 July 2000 / Accepted: 22 September 2000     

Characterization of three new S-alleles and development of an S-allele-specific PCR system for rapidly identifying the S-genotype in apple cultivars

Apple (Malus domestica Borkh), a member of the Rosaceae, shows gametophytic self-incompatibility (GSI) controlled by polymorphic S-alleles. Identifying the S-genotypes of apple cultivars can be applied on correct assignment of apple cultivars to cross-compatibility groups, which is important for the efficient production of apple fruit. This study characterized three new S-alleles (designated S ₄₄ , S ₄₅ , and S ₄₆ ) in apple and developed an efficient analysis method that can be used to characterize S-genotypes by utilizing allele-specific polymerase chain reaction rapidly. Nineteen allele-specific primers were selectively designed to identify different alleles. Using this method, S-genotypes of 157 apple cultivars were identified.     

Flowers for better pest control? The effects of apple orchard ground cover management on green apple aphids (Aphis spp.) (Hemiptera: Aphididae), their predators and the canopy insect community

Effects of habitat diversification through ground cover management on green apple aphids (Aphis spp.) (Hemiptera: Aphididae), woolly apple aphid (Eriosoma lanigerum [Haussmann]) (Hemiptera: Aphididae), their insect natural enemies and the most abundant canopy insects (in the Neuroptera, Fulgoromorpha, Cicadomorpha, Heteroptera, Coleoptera and Formicidae) were studied in an apple orchard over 6 years. The composition and diversity of the main functional groups of canopy insects was also compared. Habitat diversification was achieved by changing ground cover conditions within the orchard. In the treatment termed FLOWER, annual and/or perennial flowering plants were sown in the alleys of an apple orchard. Other ground cover treatments were weed-free bare ground (termed BAREgr) and orchard plots with alleys of mowed grass (termed GRASS), which served as control treatments. We found no evidence that habitat diversification enhanced the biological control of green apple aphids compared to the control treatments. However, the greater plant cover in FLOWER resulted in increased woolly apple aphid infestations compared to BAREgr or GRASS. The abundance of various beneficial or neutral canopy insects – Chrysoperla carnea sensu lato (Neuroptera, Chrysopidae) adults, leafhoppers and treehoppers, planthoppers, herbivorous (non-apple feeding) beetles, dipterans and parasitoid wasps – also increased in FLOWER as compared to BAREgr, with GRASS being intermediate between the other treatments. Significantly greater species richness and diversity was found in FLOWER than in BAREgr for most of the functional groups sampled, although the number of predacious insect species was similar among treatments. The composition of the studied functional groups showed high similarity in FLOWER and GRASS, but these treatments were different from BAREgr. Effects of groundcover management on the dominant insect species are discussed.     

新疆野果林苹果腐烂病病原菌鉴定及药用植物内生细菌对其抑菌效果

【背景】药用植物内生细菌能产生与寄主植物相同或相似的化合物及一些新的次级代谢产物等,具有促进宿主植物生长、抵抗病虫害、降解有毒有害化合物等作用。【目的】进一步提高苹果腐烂病生物防治的效率,丰富新疆药用植物内生细菌拮抗功能菌株的资源库。【方法】从新疆伊犁新源县和塔城额敏县野果林中采集带腐烂病病斑的果树枝条,分离鉴定苹果腐烂病病原菌,并采用平板对峙法从药用植物内生细菌中筛选对苹果腐烂病具有抑制作用的拮抗菌株。【结果】从两地共分离获得234株分离株,筛选鉴定出25株Valsa malicola和2株Valsa mali;同时,筛选出92株具有抑菌效果的内生细菌菌株,其中70株来自甘草植物内生细菌。【结论】药用植物甘草中富含较为丰富的抗苹果腐烂病病原菌的微生物菌株资源。本研究在新疆野果林苹果腐烂病的生物防治及药用植物内生细菌的开发利用等方面具有重要意义。     

福寿螺和田螺消化酶活性比较

为比较福寿螺(Pomacea canaliculata(Lamarck,1828))和当地中国圆田螺(Cipangopaludina chinensis(Gray,1832))消化能力的差异,探索福寿螺成功入侵的机制,以田螺为对照,测定了1—4龄的福寿螺和田螺的胃和肝脏的消化酶——纤维素酶(羧甲基纤维素法)、淀粉酶(3,5-二硝基水杨酸法)和脂肪酶(滴定法)的活性。结果表明:1)相同年龄的福寿螺胃和肝脏中的消化酶活性明显高于田螺。其中,纤维素酶活性分别高出1.00—2.11倍、1.66—2.84倍;淀粉酶活性分别高出1.53—3.47倍、1.47—1.80倍;脂肪酶活性分别高出2.07—4.73倍、6.13—9.93倍。2)在生长发育过程中,福寿螺胃和肝脏中的消化酶活性变化幅度(51.2%—131.2%)明显高于田螺(23.3%—47.1%)。3)福寿螺的各种消化酶之间存在协同作用。如福寿螺的淀粉酶活性与脂肪酶活性呈极显著正相关(胃中r=0.736**、肝脏中r=0.867**)。此外,胃中的淀粉酶活性还与纤维素酶活性呈显著正相关关系(r=0.696*)。相应地,田螺胃中的淀粉酶和脂肪酶之间也存在显著的正相关关系(r=0.706*),而肝脏中的纤维素酶与脂肪酶活性呈显著负相关(r=-0.593*)。4)福寿螺对纤维素类和淀粉类物质都有较强的消化能力,且能较好地消化脂肪类物质,而田螺能消化纤维素类和淀粉类物质,对脂肪的消化能力却很弱。福寿螺的纤维素酶和淀粉酶活性分别是田螺的2.42和1.88倍,脂肪酶活性达到了5.66倍。可见,福寿螺具有较高的消化酶活性,且各消化酶之间存在正协同性。这可能是导致福寿螺食量大、食性杂,使其能快速生长和成功入侵的重要原因之一。     

VARIATION IN APPLE CYTOCHIMERAS

Diploid-tetraploid cytochimeras of apple were investigated by cytological examination of buds from branches selected by the characteristics of the fruits, flowers or pollen. Eight types of cytochimeras were identified on the basis of relative size of cells, nuclei and metaphase mitoses in the apical meristem (protomeristem) and mitoses in meristematic primary tissues. There appeared to be five apical layers which contributed to the stem and four which contributed to the leaves and flower parts. Buds of various cytochimeral patterns (designated by a formula giving the ploidy of the apical layers) were found on some bearing trees propagated from known cytochimeral sources. The most frequently associated types were (a) 2–4–2–2–2 and 2–4–4–4–4 (and 2–4–2–4–4 in one cultivar), or (b) 2–2–2–4–4 and 2x. Some sports were uniformly 2–4–4–4–4. The stability of apple cytochimeras under normal conditions appeared greater in some eultivars than in others. Sprouts from severely pruned 2–2–4–4–4 trees were more variable than unpruned branches. Buds of shoots which grew from radiation-damaged buds were more variable than those from non-irradiated buds and included types not yet found by branch selection. Cytochimeral variation was interpreted to be due to layer replacement resulting from infrequent periclinal divisions in apical or axillary meristems, or from wounding of meristems by ionizing radiation.     

Mapping and functional analysis of four apple receptor-like protein kinases related to <Emphasis Type="Italic">LRPKm1</Emphasis> in <Emphasis Type="Italic">HcrVf2</Emphasis>-transgenic and wild-type apple plants

The Malus–Venturia inaequalis interaction is the most studied plant–pathogen interaction involving a woody species. Besides the cloning of an apple scab resistance gene HcrVf2, several sequences have been recently identified that are modulated after pathogen recognition in Vf-resistant genotypes. Among these, there is a putative leucine-rich repeat receptor-like protein kinase from the apple scab-resistant cv. Florina, named LRPKm1 that is induced after V. inaequalis inoculation and salicylic acid treatment. In this work, the isolation, characterization, and mapping of four new genes belonging to the LRPKm multigene family are reported. According to their cumulative expression profiles in HcrVf2-transgenic and wild-type apple plants treated with V. inaequalis, LRPKm genes have been divided in two groups. LRPKm1 and LRPKm3, giving a response related to the presence of HcrVf2, are probably involved in the recognition of pathogen-derived signals. LRPKm2 and LRPKm4, with an expression profile unrelated to the HcrVf2 gene, are putatively involved in the plant basal defense. Furthermore, we have localized LRPKm proteins at the cytological level in the plasma membrane of epidermal cells in resistant genotypes following pathogen challenge, thus confirming software predictions and molecular results. The possible involvement of LRPKm proteins in apple scab resistance and in the plant basal defense makes them attractive for a better comprehension of the molecular mechanisms of the signal transduction pathways after pathogen recognition.     

Phytoseiid mites (Acari: Phytoseiidae) on apple trees and in surrounding vegetation in southern Finland. Densities and species composition

Leaf samples were collected from sprayed (n=29) and unsprayed (n=19) apple orchards, from the surrounding vegetation (n=58) and from one arboretum (n=12), altogether from 46 plant species (1–5 samples each). The density of phytoseiid mites averaged 1.2 mites/leaf on unsprayed apple trees, but only 0.06 mites/leaf on sprayed trees. The phytoseiid density exceeded 1/leaf onAesculus hippocastani, Aristolochia macrophylla, Corylus avellana, Fragaria vesca, Frxinus excelsior, Juglans cinerea, Pterocarya rhoifolia, Ribes nigrum, Rubus odoratus, Sorbus aucuparia, S. thuringiaca, Tilia×euchlora andUlmus glabra. Other common trees and bushes inhabited by phytoseiids wereCrataegus coccinea (0.2 mites/leaf),Prunus padus (0.7),Salix caprea (0.4), andTilia cordata (0.9).Twelve species of phytoseiid mites were found, of which ten occurred on unsprayed apple trees. The most widely distributed species on apple trees werePhytoseius macropilis (in 79% of unsprayed samples),Euseius finlandicus (74%),Paraseiulus soleiger (53%),Paraseiulus triporus (37%),Amblyseius canadensis (26%) andAnthoseius rhenanus (26%). The highest densities on apple trees were found in populations ofE. finlandicus (mean 0.7 mites/leaf),Ph. macropilis (0.5) andA. canadensis (0.5). On sprayed apple trees,E. finlandicus, Pa. soleiger andPh. macropilis occurred most commonly, but their mean densities were under 0.1/leaf. Almost no phytoseiids were found in orchards sprayed with oxydemetonmethyl before blooming of apple.On other plants,E. finlandicus occurred most commonly (on 33 plant species) and in the highest densities, followed byPh. macropilis (14),Pa. soleiger (12),Pa. triporus (12) andAn. rhenanus (7).Seiulus aceri andParaseiulus talbii were identified as new phytoseiid species in Finland. It is concluded that deciduous trees and bushes in forest margins around orchards can serve as important reservoirs for phytoseiid mites, and that the dominant species in these plants would migrate into and colonize the orchards if the use of harmful chemicals were restricted.     

QTL analysis for aphid resistance and growth traits in apple

The rosy apple aphid (Dysaphis plantaginea), the leaf-curling aphid (Dysaphis cf. devecta) and the green apple aphid (Aphis pomi) are widespread pest insects that reduce growth of leaves, fruits and shoots in apple (Malus × domestica). Aphid control in apple orchards is generally achieved by insecticides, but alternative management options like growing resistant cultivars are needed for a more sustainable integrated pest management (IPM). A linkage map available for a segregating F₁-cross of the apple cultivars ‘Fiesta’ and ‘Discovery’ was used to investigate the genetic basis of resistance to aphids. Aphid infestation and plant growth characteristics were repeatedly assessed for the same 160 apple genotypes in three different environments and 2 consecutive years. We identified amplified fragment length polymorphism (AFLP) markers linked to quantitative trait loci (QTLs) for resistance to D. plantaginea (‘Fiesta’ linkage group 17, locus 57.7, marker E33M35–0269; heritability: 28.3%), and to D. cf. devecta (‘Fiesta’ linkage group 7, locus 4.5, marker E32M39–0195; heritability: 50.2%). Interactions between aphid species, differences in climatic conditions and the spatial distribution of aphid infestation were identified as possible factors impeding the detection of QTLs. A pedigree analysis of simple sequence repeat (SSR) marker alleles closely associated with the QTL markers revealed the presence of the alleles in other apple cultivars with reported aphid resistance (‘Wagener’, ‘Cox’s Orange Pippin’), highlighting the genetic basis and also the potential for gene pyramiding of aphid resistance in apple. Finally, significant QTLs for shoot length and stem diameter were identified, while there was no relationship between aphid resistance and plant trait QTLs. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effect of host plants on developmental time and life table parameters of <Emphasis Type="Italic">Amphitetranychus viennensis</Emphasis> (Acari: Tetranychidae)

The effect of host plant species including black cherry (Prunus serotina cv. Irani), cherry (Prunus avium cv. siahe Mashhad) and apple (Malus domestica cv. shafi Abadi) was studied on biological parameters of Amphitetranychus viennensis (Zacher) in the laboratory at 25 ± 1°C, 70 ± 10% RH and 16L: 8D photoperiod. Duration of each life stage, longevity, reproduction rate, the intrinsic rate of natural increase (r _m ), net reproductive rate (R ₀ ), mean generation time (T), doubling time (DT), and finite rate of increase (λ) of the hawthorn spider mite on the three host plants were calculated. Differences in fertility life table parameters of the spider mite among host plants were analyzed using pseudo-values, which were produced by jackknife re-sampling. The results indicated that black cherry might be the most suitable plant for hawthorn spider mite due to the shorter developmental period (10.6 days), longer adult longevity (25.5 days), higher reproduction (65.6 eggs), and intrinsic rate of natural increase (0.194 females/female/day). Cherry was the least suitable host plant. To determine the effect of host shifts, the mite was transferred from black cherry onto cherry and apple. In the first generation after shifting to apple, the developmental period, reproduction and life table parameters were negatively influenced. However, population growth parameters in the first generation on cherry were actually better than after three generations on this new host. This underscores the relevance of the mites’ recent breeding history for life table studies.     

X-linked heterochromatin distribution in the holocentric chromosomes of the green apple aphid <Emphasis Type="Italic">Aphis pomi</Emphasis>

Chromatin organization in the holocentric chromosomes of the green apple aphid Aphis pomi has been investigated at a cytological level after C-banding, NOR, Giemsa, fluorochrome staining and fluorescent in situ hybridization (FISH). C-banding technique showed that heterochromatic bands are exclusively located on X chromosomes. This data represents a peculiar feature that clearly contradicts the equilocal distribution of heterochromatin typical of monocentric chromosomes. Moreover, silver staining and FISH carried out with a 28S rDNA probe localized rDNA genes on one telomere of each X chromosome; CMA₃ staining reveals that these silver positive telomeres are the only GC-rich regions among A. pomi heterochromatin, whereas all other C-positive bands are DAPI positive thus containing AT-rich DNA.     

Genetics of superoxide dismutase in apple

 Four zones of superoxide dismutase (SOD), activity were detected in apple. Genetic studies confirmed the presence of three genes Sod-1, Sod-3 and Sod-4 with three, two and two alleles respectively, including one null allele. One of these genes, Sod-1, was found to be loosely linked to Prx-2 and Prx-3. The Sod-1a allele predominated in all the three groups studied, cultivars, rootstocks and Malus species. The distinct achromatic bands produced by SOD after electrophoresis facilitate its important role with peroxidase (PRX), glutamate oxalacetate transaminase (GOT) and phosphogluconate dehydrogenase (PGD) in the discrimination of apple cultivars. Received: 22 November 1996/Accepted: 29 November 1996