Testing genetic control hypotheses for Plum pox virus (sharka) resistance in apricot

The resistance/susceptibility level of 213 descendants from three different crosses between the French apricot cultivar ‘Polonais’ (susceptible to Plum pox virus, sharka), and the North American cultivar ‘Stark Early Orange’ (resistant) was evaluated during four cycles of study under controlled greenhouse conditions. Resistant:susceptible ratios were 83:17 in the case of the ‘Stark Early Orange’ open-pollination descendants, 62:38 in ‘Polonais’ × ‘Stark Early Orange’ descendants, and 28:72 in ‘Polonais’ × ‘Polonais’. These ratios were checked against the expected ratios of different genetic control hypotheses, combining one, two or three genes involved in the expression of this trait and with the resistance being a dominant (Hypothesis A) or recessive trait (Hypothesis B). In addition, two other, more complex hypotheses were proposed: that the resistance is controlled by 2 independent genes, the resistance being recessive for one and dominant for the other (Hypothesis C), and another hypothesis considering the resistance to be controlled by one dominant gene and admitting a 25% error in the evaluation process (Hypothesis D). The χ²-test was applied to compare the goodness-of-fit of the proposed hypotheses. Hypothesis D seems to be the most consistent in the three apricot crosses studied.     

Study of long-distance movement of Plum pox virus (Sharka) as an alternative resistance-evaluation method in Prunus

During the breeding programs for Plum pox virus (PPV, Sharka) resistance in Prunus, the evaluation of the new releases through symptoms observation on leaves has been contradictory and represents one of the main handicaps in these programs. In order to increase the accuracy of this traditional evaluation method, we here analyze an alternative method based on the study of the ability of a genotype to allow the long-distance movement of the virus through its vascular vessels. Two different plant models have been assayed: (a) in Model I, the inoculation was performed in the ‘GF305’ rootstock with a later grafting of the genotype under evaluation and a scion of healthy control ‘GF305’, to evaluate the long-distance movement through the studied genotype from the rootstock to the scion (xylem transport), and (b) in Model II, the inoculation with ‘GF305’ diseased scions was performed by grafting these diseased scions onto the studied genotypes, which were grafted onto healthy ‘GF305’ peach seedlings, to evaluate the long-distance movement through the studied genotype from the scion to the rootstock (phloem transport). The results show that, regardless of the presence of symptoms, susceptible genotypes allowed the movement of the virus through their vascular vessels in both directions studied. However, the resistant apricot ‘Stark Early Orange’ did not allow this movement. We propose the study of the ability of a genotype to allow the long-distance movement of the virus as an alternative and more accurate method for the evaluation of PPV resistance. However, this protocol is much more tedious than the traditional one and could be used mainly in the evaluation of a reduced number of more interesting genotypes.     

Analysis of the main factors involved in the evaluation of Prunus resistance to Plum pox virus (Sharka) in controlled greenhouse conditions

In countries like Spain or France, quarantine rules force researchers to evaluate the resistance to Sharka (Plum pox virus, PPV) in controlled, isolated conditions. This evaluation method shows important limitations resulting from the management of plants in the controlled conditions, grown in pots with artificial cycles of growth in the greenhouse and cold chamber, alternately. The objective of this study is to analyse different factors that affect the efficiency of the method of evaluation of PPV resistance in controlled greenhouse conditions. The cultivars evaluated as model genotypes were the resistant ‘Stark Early Orange’ and the susceptible ‘Real Fino’ apricot. Furthermore the ‘GF305’ peach was used as a susceptible control. The different studied factors were the inoculation protocol (rootstock or variety inoculation), the grafting success (depending on inoculation method, rootstock–variety combination and date of grafting), and the efficiency of the process in each artificial cycle of growth. Results showed that rootstock inoculation was more effective than inoculation of the variety. As rootstock, the ‘GF305’ seedlings were slightly better than the ‘Real Fino’ seedlings in the inoculation process, but they were quite similar in terms of effectiveness in the evaluation and grafting process. Grafting can be carried out in spring or autumn without having important differences. The global efficiency of the evaluation process was much higher with rootstock inoculation.     

Early detection of graft incompatibility in apricot (Prunus armeniaca L.) using phenol analyses

The study investigated the role of phenols in apricot graft incompatibility. Assays of phloem with cambium from 1-year-old apricot trees of cultivars Marlen, Leskora and Betinka which were grafted on the rootstocks of different genetic origin: M-LE-1, Lesiberian, MY-KL-A, Tetra, Penta, Green Gage, Julior, MRS 2/5 and Isthara were analysed with HPLC (together 23 scion/stock combinations). The phloroglucinol, catechin, p-coumaric acid and further non-identified phenols with the retention time 23–25 and 30 min were determined. The content of individual phenol compounds was related to specific cultivar/rootstock combination. The minimum number of statistical significant differences in the phenol content between tissues above and below graft union was established in homospecific combinations (P. armeniaca/P. armeniaca). Cultivars Marlen, Leskora and Betinka differ in the degree of compatibility or incompatibility with rootstocks. The pattern of non-identified phenol 23 in different graft combinations is similar to catechin and p-coumaric acid.     

Cross-transferable polymorphic SSR loci in Prunus species

This work reports the transferability and polymorphism of previously reported SSRs in 10 Prunus species. The availability of a large number of SSRs in the genus Prunus makes marker choice random, while preventing comparison of results in fingerprinting studies. The availability of SSR markers, polymorphic in a wide sample of Prunus species, would facilitate marker choice, while allowing the comparison of results. In this work, microsatellite markers useful for analyzing 10 different Prunus species (P. persica, P. dulcis, P. armeniaca, P. domestica, P. insititia, P. salicina, P. cerasifera, P. avium, P. cersus and P. mahaleb) were searched through screening SSRs previously reported to be conserved and/or polymorphic in more than one Prunus species. A selected group of 13 SSRs, transferable to the 10 species, was analyzed in terms of their usefulness for analyzing these species. The amplification range, polymorphism and variability detected by these loci are reported. The information provided will be useful for Prunus genetic studies as well as conservation and management of Prunus germplasm resources.     

Effect of climatic conditions on the overcoming of dormancy in apricot flower buds in two Mediterranean areas: Murcia (Spain) and Tuscany (Italy)

For temperate-zone fruit species such as apricot, when winter cold requirements are not adequately satisfied, negative repercussions on productivity occur. The aim of this research was to study the effect of, mainly temperature on the overcoming of dormancy in different apricot cultivars growing in two areas (Italy, Tuscany: lat. 43°02′N, long. 10°36′E; Spain, Murcia: lat. 38°16′N, long. 1°16′W) representative of the Mediterranean climate. Trials were conducted for two consecutive years on the same genotypes: ‘Currot’, ‘San Castrese’, ‘Goldrich’, ‘Stark Early Orange’ and ‘Orange Red’. These genotypes cover the range of chilling requirements (CR) in the apricot species in these Mediterranean areas. The CR (measured by chill units, CU and portions) for breaking of dormancy, heat requirements, flowering date and flowering and fruit-set percentages were measured. Temperatures were recorded and transformed into the corresponding CU and portions by the Utah Model and the Dynamic model, respectively. The winter climatic conditions determined a dissimilar chill unit accumulation in Tuscany (Italy) and Murcia (Spain) as well as an important effect of the year in both areas. While all cultivars with the exception of Stark Early Orange overcame dormancy, significant differences regarding the CR of cultivars for two different years and growing in two environments were observed, depending also on the cultivar. The results showed that both the Utah Model and Dynamic Model were not completely accurate with regard to establishing the CR for dormancy release under a Mediterranean climate. Temperature should be analysed together with other climatic factors in order to improve the CR assessment.     

Effect of carbohydrates on in vitro low-temperature storage of shoot cultures of apricot

Procedures for cold storage of in vitro cultures can delay subculturing, reducing production costs and risks of contamination and somaclonal variation. The present work investigates the effects of media with sorbitol (116.8 mM, medium SO) or sucrose (58.4 mM) alone (medium SU), or the latter in combination with mannitol (58.4 mM, medium M) on 7-month storage at 5 °C of apricot shoots, cv San Castrese and Boreale. Shoots in SO survived in lower percentages and grew less than in the other treatments during storage, and died in large numbers after transfer to standard culture conditions. In comparison to other treatments, survival was 100% in the presence of M and both shoot weight and number of surviving proliferated axillary shoots was increased. Moreover, M improved regrowth compared to SU under standard culture conditions. The SOD and CAT activity confirmed the higher stress of shoots stored in SO than controls, and in contrast, the low stress of shoots in M.     

Genetic diversity of Rehmannia glutinosa cultivars based on sequence-related amplified polymorphism markers

SRAP analytic system was used to assess genetic diversity of Rehmnnia glutinosa. Twenty-three Rehmnnia glutinosa cultivars were screened with 288 primer combinations, of which 13 produced stable and reproducible amplification patterns in three repetitive experiments. Among a total of 338 amplified fragments, 306 (90.5%) were polymorphic, with an average of 23.5 fragments for each primer combination. The percentage of polymorphic bands for each primer combination varied from 58.3 to 100%. The cultivars had a similarity ranging from 0.335 to 0.713 with a mean of 0.518. Shannon's diversity index and expected heterozygosity were 0.3217 and 0.2008, respectively. Based on the cluster, which were conducted on the similarity matrix of SRAP marker data, the cultivars were divided into four groups at the 20 rescaled distance cluster combine. The results demonstrated that SRAP is a stable marker technique for the assessment of genetic diversity of Rehmannia glutinosa cultivars, and that the level of genetic diversity among them from different production areas was relatively high.     

Genetic diversity of apricot revealed by a set of SSR markers from linkage group G1

Eight polymorphic simple sequence repeat (SSR) markers located in the G1 linkage group of apricot (Prunus armeniaca L.) were previously developed and evaluated in a small set of cultivars. Those primers were used for studying variability in 77 apricot cultivars belonging to five different geographical groups, such as Chinese, Asian (Irano-Caucasian and Central Asian), North American, Mediterranean and Western European as well as Middle European cultivars. Six of the markers were polymorphic and revealed a total of 71 alleles ranging from 5 (aprigms11) to 20 (aprigms1) alleles per locus with a mean value of 11.83 alleles per locus. In conclusion, the SSR loci located in the G1 linkage group show a level of polymorphism which is similar to loci dispersed throughout the entire genome. The total number of alleles and the number of unique alleles were the highest in Chinese apricots and the lowest in Middle European cultivars. Heterozygosity also showed a decrease from Asia and China to Middle Europe. No association could have been observed between any SSR markers tested and plum pox virus (PPV) resistant phenotype of cultivars. PPV resistant cultivars did not form a separate clade on the dendrogram obtained by UPGMA cluster analysis. Middle European and Chinese cultivars formed separate clusters while other genotypes formed smaller multiple sub-groups or scattered among different clusters. Our results support previous hypotheses on the origin of PPV resistance in North American apricots. The allele data was also presented in a form that allowed the easy observation of allele frequencies in each geographical group at each locus. Using this data field, differences and similarities between cultivar groups can be easily assessed. The analysis demonstrated the links between the North American and Mediterranean apricot germplasm and confirmed that the Chinese and Eastern European cultivars are distantly related.     

Reaction of cucurbits species to infection with Zucchini lethal chlorosis virus

Zucchini lethal chlorosis virus (ZLCV) is a tospovirus that infects species of cucurbits in Brazil and is transmitted by the thrips Frankliniella zucchini. The purpose of the present work was to evaluate the reaction of seven species of cucurbits to infection with ZLCV under field and greenhouse conditions in Piracicaba County, SP. In the field experiment, ZLCV infection occurred naturally. In the greenhouse, plants were mechanically inoculated with ZLCV at the cotyledonal stage. Evaluations were based on symptoms expression and detection of the virus by PTA-ELISA. The percentages of infected plants for field and greenhouse assays, respectively, are indicated in parenthesis for each species/cultivar: Cucurbita pepo var. Caserta (72.9 and 70.7); C. maxima var. Alice (16.0 and 10.4); C. maxima var. Exposição (0.0 and 0.0); C. moschata var. Menina Brasileira (29.2 and 18.2); C. maxima × C. moschata hybrid Takaiama (63.4 and 42.7); Citrullus lanatus var. Crimson Sweet (25.0 and 25.0); Cucumis sativus var. Safira (14.3 and 41.2); and C. anguria (21.4 and 22.7).     

Comparative analysis of genetic diversity in Tunisian apricot germplasm using AFLP and SSR markers

Eighty-one accessions representing apricot germplasm in Tunisia were collected from different areas of cultivation and fingerprinted using amplified fragment length polymorphism (AFLP) and microsatellites (SSR) markers. A total of 339 polymorphic markers were revealed using 5 AFLP primers combinations and 24 SSR loci. AFLP and SSR markers expressed a high level of polymorphism allowing the distinction of the accessions with an efficiency coefficient of discrimination of 100% for AFLP and 97% for SSR markers. Genetic diversity structure was assessed with AFLPs and SSRs markers separately then with combined matrix data by the help of hierarchical clustering elaborated using Wards method based on Nei and Li (1979) distances. Comparison of the obtained dendrograms revealed a phylogeographic structure into two major groups with significant conservation between the observed subgroups in relation with the geographic origin of the accessions. The relative efficiency of the markers in determining the genetic relationships among apricot accessions has been assessed and a combination of AFLPs and SSRs markers was the most effective. In addition, Mantel test based on genetic distances indicated highly significant correlation between AFLP-SSR data and each of the AFLP and SSR ones, with Pearson correlation values of r = 0.873 and r = 0.692, respectively, revealing the higher efficiency of the combination of both molecular techniques (AFLP and SSR) to estimate the levels of genetic variability among apricot germplasm.     

Models for predicting the mass of apricot fruits by geometrical attributes (cv. Shams,Nakhjavan, and Jahangiri)

Fruits with the similar weight and uniform shape are in high demand in terms of marketing value. Therefore, an awareness of grading fruit based on weight is crucial. A part of this research was aimed to present some physical properties of three Iranian apricot cultivars (Shams, Nakhjavan, and Jahangiri). In addition, apricot mass was predicted by different physical characteristics with linear and nonlinear models as three different classifications: (1), single or multiple variable regressions of apricot dimensional characteristics. (2), single or multiple variable regressions of apricot projected areas and (3), estimating apricot mass based on its volume. All properties considered in the current study were found to be statistically significant at the 1% probability level. The highest and the lowest dimensional characteristics were found for Jahangiri and Nakhjavan cultivars, respectively. Sphericity values had significant difference among the tested cultivars. The latter property values were 0.971, 0.917, and 0.973 for Shams, Nakhjavan, and Jahangiri cultivars, respectively. Based on the results, the surface area of the Jahangiri cultivar was found to be 6458.35 mm², followed by the Shams and Nakhjavan cultivars, which had a mean of 6115.36, and 4395.25 mm², respectively. In this paper, the coefficients of static friction of the cultivars on three different surfaces are also reported. The results showed that mass modeling of apricot based on minor diameter and three projected areas are the most appropriate models in the first and the second classifications, respectively. In third classification, the best model was obtained on the basis of the actual volume as M = 0.997V_m + 0301, R² = 0.98, R.S.E. = 1.711 with R² = 0.98. It was concluded that the suitable grading system of apricot mass is based on minor diameter as nonlinear relation: M = 0.0019c^2.693, R² = 0.96, R.S.E. = 2.2.     

Production protocol development for greenhouse cut Linaria, Lupinus, and Papaver flowers

Linaria maroccana Hook. f. Ann., ‘Lace Violet’, Lupinus hartwegii ssp. cruikshankii Lindl. ‘Sunrise’ and Papaver nudicaule L. ‘Meadow Pastels’ seeds were directly sown into 105 cell plug trays and received either ambient light or supplemental high intensity discharge (HID) lighting. For each species, a 2 × 3 × 3 factorial was used with two light intensities during propagation, three transplant stages, and three night temperatures. Seedlings were transplanted at the appearance of 2–3, 5–6, or 8–9 true leaves. Transplanted Linaria and Papaver seedlings were placed at 5/11, 10/16, or 15/21 ± 1 °C night/day temperatures and Lupinus seedlings were placed at 15/24, 18/25, or 20/26 ± 2 °C night/day temperatures. For this study, the optimum production temperature for Linaria was 10/16 °C as the cut stems produced at 15/21 °C were unmarketable and production time was excessively long at 5/11 °C. At 10/16 °C, Linaria seedlings should be transplanted at the 2–3 leaf stage to maximize stem number, stem length and profitability. For Lupinus the optimum temperature was 15/24 °C due to long stems and high profitability per plant. Lupinus seedlings should be transplanted at the 2–3 leaf stage when grown at 15/24 °C to obtain the longest and thickest stems; however, $/m² week was higher for plants transplanted at the 8–9 leaf stage due to less time in finishing production space. For Papaver, the 15/21 °C temperature was optimal as that temperature produced the longest stems in the shortest duration, resulting in the highest $/m² week. At 15/21 °C Papaver plants should be transplanted at the 2–3 leaf stage. Supplemental HID lighting had no effect on any of the species.     

The responses of photosynthetic capacity,chlorophyll fluorescence and chlorophyll content of nectarine (Prunus persica var. Nectarina Maxim) to greenhouse and field grown conditions

Three nectarine (Prunus persica var. Nectarina Maxim.) cultivars grown under solar-heated greenhouse and open-field in northwest China, were tested to evaluate their photosynthetic and chlorophyll fluorescence response to both growth conditions, and whether nectarine plants acclimate to the solar-heated greenhouse growth condition. Comparisons of light-saturated photosynthetic capacity (Amax) and CO₂-saturated photosynthetic capacity (RuBPmax) indicated that each cultivar (Z, Zao-Hongzhu; H, Hua-Guang; Y, Yan-Guang) maintained similar rates of light-saturated and CO₂-saturated carbon assimilation when grown in both conditions. The curve of diurnal variation of net photosynthetic (P_N) rate showed double peaks in open-field but single when grown in greenhouse. Compared with open-field-grown plants, a significant increase of daily average P_N was found in Z but decreased in Y in greenhouse. The diurnal variation of F_v/F_m indicate that plants grown in greenhouse experience less photoinhibition than in open-field condition. A reduction in chlorophyll (chl) a/b ratio in leaves of greenhouse grown plants with significant increase in chlorophyll (chl) b content were obtained. The results suggest that some nectarine cultivars have the ability to acclimate to the solar-heated greenhouse growth condition.     

Chemical compositions and insecticidal effects of essential oils isolated from Achillea gypsicola, Satureja hortensis, Origanum acutidens and Hypericum scabrum against broadbean weevil (Bruchus dentipes)

The essential oils of aerial parts of Achillea gypsicola Hub.-Mor., Hypericum scabrum L., Satureja hortensis L., and Origanum acutidens (Hand.-Mazz.) Letswaart were analyzed in this study by GC and GC–MS and their oils were tested for toxicity against broadbean weevil (Bruchus dentipes). A. gypsicola oil contained camphor (40.17%), 1,8-cineole (22.01%), piperitone (11.29%), borneol (9.50%) and α-terpineol (1.56%) as major components. A total of 74 components were identified by GC–MS in H. scabrum oil, including α-pinene (9.26%), terpinen-4-ol (5.12%), camphor (5.94%), δ-cadinene (4.52%), pulegone (4.45%), γ-muurolene (4.12%), pinocarvone (3.97%) and β-caryophyllene (3.42%) as predominant components. The essential oils of O. acutidens and S. hortensis were characterized by high contents of carvacrol (86.99% and 55.74%), γ-terpinene (0.71% and 20.94%), p-cymene (1.95% and 12.30%), α-terpinene (0.13% and 2.04%) and β-caryophyllene (1.30% and 1.08%). All of the essential oils were toxic to adults of B. dentipes and insect mortality increased with increasing concentration of each oil. The oils (20 μl dose) brought about 100% mortality in 36 h. Although desirable insecticidal activities against the pest were achieved with the oils from all four plant species, S. hortensis and O. acutidens oils were more effective, particularly after 6 h of treatment. The current results concluded that the essential oils, in particular O. acutidens and S. hortensis oils, may be used as potential botanical insecticides against B. dentipes.     

Taxonomic revision of Dendrobium moniliforme complex (Orchidaceae)

Taxonomic revision of Dendrobium moniliforme complex is presented. D. moniliforme complex is characterized by the even slim stems, bracts with brownish zone, semi-spherical anther cap and the hairy disc of lip. Dendrobium tosaense, Dendrobium officinale and Dendrobium guangxiense were excluded by having membranous bracts lacking brownish zone, anther cap conical and bifid. Two species are recognized in this complex, i.e., D. moniliforme and Dendrobium wilsonii. D. wilsonii differs from D. moniliforme by having elliptic leaves about 1.3–2 cm wide, dorsal sepal 3.0–4.0 cm long, 0.6–0.9 cm wide, petals elliptic to oblong, 3.0–4.0 cm long, 1.0–1.5 cm wide, lip elliptic to ovate–lanceolate, 2.6–3 cm long, 1.2–1.5 cm wide.     

Assessment of genetic diversity of Latvian and Swedish sweet cherry (Prunus avium L.) genetic resources collections by using SSR (microsatellite) markers

Three previously described highly polymorphic SSR (microsatellite) primer pairs were tested on 126 sweet cherry (Prunus avium L.) accessions to adapt a fast, reliable method for preliminary screening of sweet cherry germplasm collections and to compare two sweet cherry germplasm collections: at the Latvia State Institute of Fruit-Growing, Dobele (LIFG-Dobele) and at the Division of Horticultural Genetics and Plant Breeding at Balsgård, Department of Crop Sciences, Swedish University of Agricultural Sciences (SLU-Balsgård). The SSR loci were highly polymorphic with 4–10 different alleles and 5–18 genotypes. Heterozygosity values ranged from 0.431 to 0.809, gene diversity (PIC) values ranged from 0.400 to 0.753, and the discriminating power of each locus varied from 0.631 to 0.894. The combined discriminating power of all loci was highly effective (0.996). Sixteen identical accession groups with the same allele profile were discovered in both collections. This study demonstrated that SSR fingerprinting with the three primer pairs tested, can be used for preliminary characterization of sweet cherry germplasm collections.     

Phenotypic variability and genetic structure in plum (Prunus domestica L.), cherry plum (P. cerasifera Ehrh.) and sloe (P. spinosa L.)

In the present study, phenotypic variability of 80 plum (Prunus domestica L.) varieties maintained in the French National Plum Collection was evaluated with 19 quantitative traits. In addition, genetic diversity and genetic structure was studied in three plum species (P. domestica L., Prunus cerasifera Ehrh. and Prunus spinosa L.) using chloroplast DNA (cpDNA) markers and five single sequence repeat (SSR) loci. Based on phenotypic traits, some varieties, such as mirabelle plums, grouped together. Bayesian structure analysis was used to identify different genetic groups, whereby damson plums were clearly distinguished from greengage plums. When examining the three species together, a higher level of cpDNA allelic richness was found in P. cerasifera and in P. spinosa than in P. domestica where only five cpDNA haplotypes were detected in the national plum collection, with one main haplotype that accounted for 80% of the varieties studied. P. domestica cpDNA haplotypes tended to group together with P. cerasifera haplotypes whereas most of P. spinosa haplotypes formed a separate cluster. SSR markers were somewhat able to distinguish the three species. These results provide some clues as to the origin of plum and the various plum varieties. Our results also provide useful information for the management of plum genetic resources.     

Syrian pear (Pyrus syriaca) as a pollinator for European pear (Pyrus communis) cultivars

In Israel four European pear cultivars are grown: ‘Spadona’ is the main cultivar and ‘Coscia’, ‘Gentile’ and ‘Spadochina’ are its pollinators. However, molecular S-genotyping revealed that ‘Spadona’ is semi-compatible with its three pollinators. This explains, at least in part, the relatively low pear yield in Israel. The Syrian pear (Pyrus syriaca) grows wild in Israel and blooms intensively, overlapping the blooming of the cultivated European pears. Cross-fertilization between Syrian pear and ‘Spadona’ was shown to be efficient suggesting that Syrian pear might be a potent pollinator for ‘Spadona’. Twenty-six Syrian pear seedlings, from different sites in north-east Israel were S-genotyped identifying 11 that are fully compatible with the four European pear varieties cultivated in Israel. By this screening, 24 different S-RNases were cloned; ten of them are new, whereas the other fourteen had been identified previously. In addition, seedlings of two wild pear species were also S-genotyped. Two seedlings from Pyrus betulifolia and one from Pyrus korshinskii were found to be genetically compatible with the four European pear cultivars. From these seedlings four S-RNases were cloned, two are new, one had been cloned previously and one was identical to an S-RNase allele cloned from Syrian pear in this work.     

Pome fruit viruses at the Canadian Clonal Genebank and molecular characterization of Apple chlorotic leaf spot virus isolates

A survey for apple and pear viruses was carried out at the Canadian Clonal Genebank (CCG), Harrow, Ontario, Canada, during the fall/winter of 2007 and spring of 2008. Leaves and/or dormant cuttings were randomly collected from 438 to 122 accessions of apple and pear, respectively. Samples were tested by Double Antibody Sandwich-Enzyme Linked Immunosorbent Assay (DAS-ELISA) for the presence of Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV) and Apple mosaic virus (ApMV). Infection rates for apples were ACLSV (48.1%), ASGV (10%), ASPV (6.6%) and ApMV (7.1%), and for pears ACLSV (42.6%). ACLSV was detected and characterization by multiplex RT-PCR with primers targeting a fragment of 677 bp corresponding to the partial coat protein (CP), movement protein (MP) and untranslated (3′UTR) region in 22 accessions of apple and pear. Multiplex RT-PCR showed a higher sensitivity over the ELISA test. The nucleotide and amino acid deduced partial CP identities ranged from 82.6–100% to 91–100%, respectively, while partial MP identities was 62.5–100% at aa level based on the amplified fragment appropriate for partial MP using a frame shift, among 22 ACLSV isolates. Phylogenetic analyses based on the partial CP region clustered CCG ACLSV isolates in two different groups, while those based on the partial MP region embraced CCG ACLSV isolates in two sub-clusters within the same group. This is the first report on the detection of ACLSV, ASPV, ASGV and ApMV at CCG, and the molecular characterization of ACLSV isolates in apple and pear plants from worldwide countries to deduce possible heterogeneity and evolution.