Top working method and bloom density of pollinizers as productive determinant for spur type apple (Malus x domestica Borkh.) cultivars

Apple (Malus x domestica Borkh.) shows gametophytic self incompatibility and requires cross pollination by suitable pollinizers. Gradual decline in natural pollinators and insufficient proportion of pollinizers have been observed in the apple orchards over the years across the North Western Himalayan region. This situation resulted in considerable pollination problem in the region. The present study was undertaken (during the years 2006–2010) to examine the effect of top working methods and time as well as type of pollinizers on different growth parameters for efficient pollination management. Chip budding performed in fall, late winter and summer gave significantly higher success rate (90.0, 89.8 and 80.9%, respectively). Other methods viz., side rind graft (85.6%) and cleft graft (71.2%) during February–March, and T-bud (71.7%) during July–August also gave better success. The highest shoot numbers (4.1) and length (92.8 cm) were recorded when chip budding was performed during September–October and February–March, respectively. In 4th year, chip budded scion branches of all the pollinizers recorded significantly higher number of spurs/m twig length with a range of 22.8 (December–January) to 24.7 (September–October). Significantly the highest bloom density (22.1) was recorded on chip budded (February–March) branches of pollinizers in 4th year. Fruit set on top worked trees of ‘Oregon Spur’ was only in the range of 18.0–20.1% in 1st year and reached to the range of 35.6–41.5% in 4th year. Seed numbers/fruit also increased to the range of 7.3–7.9 in 4th year from 2.9 to 4.6 in 1st year. All these factors resulted in better yield efficiency (2.6–3.8) of ‘Oregon Spur’ in 4th year. ‘Manchurian’ crab produced significantly higher shoot length of 86.5–87.2 cm and 146.1–149.2 cm in 1st and 4th years, respectively. However, ‘Stark Spur’ produced the highest number of spurs/m twig length of 28.9 when grafted/chip budded during December–January or February–March, and 27.1 when chip/T-budded during September–October or July–August. Top working with ‘Manchurian’ crab also resulted in significantly higher fruit set (44.2–45.4%) and yield efficiency (3.6–3.8) on ‘Oregon Spur’ in 4th year. This was followed by ‘Stark Spur’ as pollinizer (37.1–38.2% and 2.9–3.1, respectively). Highly significant positive correlation of shoot numbers, spur density and bloom density of pollinizers with fruit set and yield efficiency of ‘Oregon Spur’ were observed. ‘Manchurian’ crab was found to be very efficient as pollinizer, followed by ‘Stark Spur’ for spur type ‘Oregon Spur’ apple cultivar on the basis of higher bloom density and fruit set parameters.     

Detecting different levels of drought stress in apple trees (Malus domestica Borkh.) with selected biochemical and physiological parameters

Rational irrigation scheduling based on sensing drought stress directly in plants is becoming more important due to increasing worldwide scarcity of fresh water supplies. In order to evaluate a set of potential biochemical and physiological stress indicators and select the best drought stress markers in apple trees, two experiments with potted trees and an experiment with intensive orchard grown apple trees ‘Elstar’ and ‘Jonagold Wilmuta’ were conducted in early summer in tree following years. Biochemical parameters: ascorbic acid, glutathione, tocopherols, chlorophylls, carotenoids, free amino acids, soluble carbohydrates, and physiological parameters already known as stress indicators in apple trees: predawn and midday leaf water potential, net photosynthesis (Pn), stomatal conductance (g_s), transpiration (Tr) and intercellular CO₂ concentration (Ci) were measured in leaves of apple trees subjected to different intensities of slowly progressing drought or no drought. Our study pointed out zeaxanthin and glutathione as the best drought stress markers in apple trees. Ascorbate and sorbitol appeared to be reliable indicators of moderate drought only. Responses of other tested biochemical parameters were not consistent enough to prove their role as drought stress markers in apple trees. Relative air humidity should be taken in consideration when physiological parameters g_s, Pn, Tr and Ci are used as drought stress markers in apple trees. Our study revealed that in situations where low relative air humidity affects g_s and with g_s connected physiological parameters, biochemical markers may be better tool for determination of drought stress intensities in apple trees.     

Physico-chemical and antioxidant properties of six apple cultivars (Malus domestica Borkh) grown in southern Brazil

The objective of this study was to compare the physico-chemical properties and antioxidant activity of six apple cultivars grown in southern Brazil. Apple peel color, dry matter, total soluble solids, pH, total sugars, titratable acidity, total phenolics, total monomeric anthocyanin and total antioxidant activity were measured in the apple cultivars Imperatriz, Daiane, Fred Hough, Fuji Suprema, Galaxy and Baronesa. The results showed great quantitative differences in the composition of the apple cultivars. Of all the cultivars, the peel of Galaxy was slightly more red-colored and that of Fred Hough was the least red-colored. The dry matter varied from 15.24% (Galaxy) to 19.55% (Fuji Suprema), the soluble solids content was between 11.8 (Fred Hough) and 14.0 (Daiane) °Brix, pH values varied from 3.90 (Imperatriz) to 4.27 (Fred Hough), the total sugar content (g 100 g⁻¹) ranged from 11.54 (Imperatriz) to 14.78 (Fuji Suprema) and the titratable acidity content (g 100 g⁻¹) varied from 0.20 (Baronesa) to 0.36 (Imperatriz). The total phenolic content (GAE 100 g⁻¹ fresh matter) observed in the apple cultivars was between 105.4 (Baronesa) and 269.7 mg (Imperatriz). The values of the total anthocyanin content (mg 100 g⁻¹ FM) ranged from 4.79 (Fred Hough) to 41.96 (Galaxy). The highest total antioxidant activity was observed in Imperatriz (739 μmol TEAC 100 g⁻¹ FM), while the lowest value was found in Fuji Suprema (335 μmol TEAC 100 g⁻¹ FM). There was a strong correlation between total monomeric anthocyanin content and all peel color measurements and between antioxidant activity and total phenolic content. The results suggested that genotype is the main factor that determines the composition of bioactive compounds in apples and this provides important information on how to make the best use of the apple cultivars investigated.     

Improving the performance of calcium-containing spray formulations to limit the incidence of bitter pit in apple (Malus x domestica Borkh.)

Laboratory and field experiments were carried out with apples (Malus x domestica Borkh.) cv. ‘Golden Reinders’, to assess the efficacy of sodium salt of carboxymethyl ether of cellulose (0.5%, CMC) as an adjuvant for Ca spray formulations containing either Ca-chloride or Ca-propionate as active ingredient (120 or 250 mM Ca). This additive significantly increased the retention of Ca-containing solutions by the apple skin and prolonged the process of drying of the solution at room temperature. Four days after immersion of apples in 0.5% CMC plus CaCl₂ or Ca-propionate solutions (120 and 250 mM Ca) significant Ca increases were recorded in the peel and cortex of treated fruits. Application to apple trees of in-season sprays containing 250 mM CaCl₂ plus 0.05% Tween 20, Ca-propionate (120 and 250 mM Ca) plus 0.5% CMC or 250 mM CaCl₂ plus 0.5% CMC had no impact on fruit yield and quality, but significantly limited the rate of bitter pit incidence during the following 3-month cold-storage period. Evidence is provided that addition of appropriate adjuvants to Ca sprays can favour the distribution of Ca into the apple fruit and helps to reduce the incidence of Ca-related disorders over the postharvest cold-storage period.     

Reduction of apple tree height (Malus domestica Borkh) cv. Ultra Red Gala/MM111 does not decrease fruit yield and quality

In Chile, like in other countries, a high percentage of apple (Malus domestica Borkh) orchards are grafted on vigorous or semi-vigorous rootstocks. The need to decrease the amount of labor involved and increase efficiency has motivated this study on the effect of reducing the height of cv. Ultra Red Gala/MM111 trees in a commercial orchard in the Maule Region of Chile. Apple trees were planted in 2003, and their tree heights were adjusted to 2.5, 3.0 and 3.6 m prior to blooming in 2006. The reduction of plant height from 3.6 m to 2.5 m determined a significant reduction in canopy volume (26% and 29% in 2007/2008 and 2008/2009, respectively), but no differences were detected in leaf area index (LAI) and the photosynthetic active radiation (PAR) transmitted through the canopy. However, a higher proportion of the canopies of the shorter trees had over 30% of incident PAR, the threshold for the production of good fruit quality and flower-bud differentiation. There were no differences in fruit production among plant heights, accumulating 131 tonnes ha⁻¹ in the three growing seasons. Over the three seasons, the shortest plants (2.5 m) required 19, 57 and 42% less labor time at harvest, respectively, than the plants maintained at a height of 3.6 m. There were no evident differences in fruit quality among the treatments; flesh firmness varied between 73.0 and 74.0 N, soluble solids between 11.6 and 12.6°Brix, mean weight between 180 and 200 g, and the percentage of red coloring exceeded 59% of the Premium fruit in the three growing seasons. Our results suggest that it is possible to manage plants with reduced height on semi-vigorous rootstocks and thus reduce the time necessary for harvesting, without affecting fruit yield and quality.     

Phytochemical and antioxidant properties of selected fig (Ficus carica L.) accessions from the eastern Mediterranean region of Turkey

Fig has been a typical fruit component of the health-promoting Mediterranean diet for a very long time. Phytochemical characters and antioxidant capacity of green-, yellow-, brown-, purple-, and black-fruited fig (Ficus carica L.) accessions were investigated. In this study, total phenolics (TP), total anthocyanins (TA), fructose (FRUC), glucose (GLUC), sucrose (SUC), and variables (such as L*, a*, C*, and hue°) describing fruit skin colors were examined. Also, the antioxidant capacity (TAC) of fig fruits was determined by the ferric reducing antioxidant power (FRAP) assay. Antioxidant capacity was significantly correlated with the polyphenol and anthocyanin (r = 0.74 and 0.63, respectively) contents of fruits. Black fig accessions had the highest TAC (range of 7.9–16.1, mean 12.4 Fe²⁺ mmol/kg FW), TA (range of 32.3–356.0, mean 128.4 μg cy-3-rutinoside/g FW), and TP content (range of 69.1–220.0, mean 118.9 mg GAE/100 g FW). These black-fruited accessions had 2-fold greater TAC, 15-fold greater TA, and 2.5-fold greater TP than green and yellow fig accessions. However, the FRUC, GLUC, and SUC content of brown and purple fig accessions were higher than those of other color groups. The predominant sugars present were fructose (∼56%) and glucose (∼43%), as determined by HPLC.     

Arginine metabolism in roots and leaves of apple (Malus domestica Borkh.): The tissue-specific formation of both nitric oxide and polyamines

In addition to serving as an important nitrogen reserve, l-Arginine participates in various physiological processes in plants. In plants, like in animals, nitric oxide (NO) is synthesized through nitric oxide synthase (NOS) pathway, while polyamines (PAs) are through arginine decarboxylase (ADC) and/or arginase–ornithine decarboxylase (ODC) pathway in arginine metabolism. In this study, arginine metabolism in fine roots and leaves of different development stages (white roots, brown roots, young leaves and mature leaves) were investigated, using potted 3-year-old apple plants. The results showed that different pathways in arginine metabolism are positively correlated, producing polyamines and nitric oxide in a tissue-specific manner. Higher levels of arginine, polyamines and nitric oxide, as well as arginase, ADC, ODC and NOS activities exist in roots than in leaves. White roots and young leaves exhibit higher levels of arginine metabolism than brown roots and mature leaves, respectively. As both polyamines and nitric oxide are multifunctional molecules involved in plant development and stress response, it is possible to conclude that the unique physiological roles of l-Arginine in plants might be associated with the coordinated biosynthesis of both polyamines and nitric oxide via arginine metabolism.     

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.     

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.     

Cultivar variation in apple peel and whole fruit phenolic composition

The quali-quantitative distribution of phenolic compounds varies considerably between apple flesh and peel, but the concentration of phenolics is substantially higher in the peel than flesh. Because the peel comprises only a small percentage of the entire fruit weight, its significance as a donor of phenolics is disputable. We assessed the contribution of the peel to the total phenolic yield of 19 apple cultivars. Calculations were based on the weight of the whole fruit and the peel (which is frequently discarded) and the concentration of individual phenolic compounds. On average, 8, 24, 32, 50 and 66% of chlorogenic acid, (+)-catechin, (−)-epicatechin, phloridzin, and rutin, respectively, were present in the peel, which constitutes about 6–8% of the whole apple weight. With the exception of chlorogenic acid, 50% or more, on average, of the above phenolics were present in the peel of ‘Granny Smith’, ‘Idared’, ‘Red Rome’, ‘Jonamac’ and ‘Gloster’ apples; the highest percentage was found in ‘Starking Delicious’ apple peel (82%). The lowest peel contribution to total phenolic content per whole apple ranged between 26 and 29% and was observed in ‘Pilot’, ‘McIntosh’ and ‘Prima’ apples. Presented results may be useful for further investigations of the relationship between phenolics and agronomical parameters or future selection of apple genotypes having improved nutritional quality when consumed as fresh or as processed apple products.     

Improvement of quality and storability of apple cv. Aroma by adjustment of some pre-harvest conditions

To improve the quality of the important Swedish apple cultivar Aroma and increase its resistance to bruising and storage decay, three studies were conducted at Kivik research station during the period 1998–2003. The effects of tree age, pruning time and nitrogen fertilization were investigated. Fruits from young trees (younger than 6 years) had a lower resistance to bruising and Pezicula malicorticis decay, while fruits from trees older than 20 years seemed to show a quality and storability decline. Choosing a suitable summer pruning time was an effective factor for fruit storage potential. Thinning cut during August (5–6 weeks before harvesting) improved fruit colour and decreased decay without negatively influencing tree yield. Applying fertigation according to fruit development phase and soil nitrogen content had a very important influence on fruit quality and storability. Appropriate nutrition supply produced more deeply coloured fruit that maintained their quality after storage for 4 months. The bruising sensitivity of cv. Aroma apples was correlated with their N and K/Ca content. More coloured fruits showed less sensitivity to bruising.     

Genetic transformation of Prunus domestica L. using the hpt gene coding for hygromycin resistance as the selectable marker

The study and development of transformation technology with new selection schemes is important for various fundamental studies and for crop trait improvement via genetic engineering. Here we have shown that hygromycin resistance is an effective system for plum genetic transformation. Embryonic axes of mature seeds were co-cultivated with Agrobacterium strain LBA4404 containing the pC1381 plasmid carrying the hygromycin phosphotranferase gene (hpt) and β-glucuronidase (GUS) gene or with strain EHA105 containing the plasmid pC1301 carrying the same marker and reporter genes. The latter strain containing a pC2301 plasmid carrying the neomycin phosphotransferase gene (nptII) gene was used as a control. Infected explants were placed on shoot induction medium containing either 5 mg L⁻¹ hygromycin or 75 mg L⁻¹ kanamycin for selection. Green shoots developed from the explants under hygromycin pressure. These shoots showed continued and vigorous growth and development upon transfer onto fresh hygromycin medium. PCR using hpt sequence primers, and Southern blot analysis using a probe from the hpt gene, confirmed the presence of the transgenes and their stable integration in regenerated plants. Full transgenic plants were obtained in a greenhouse. Hygromycin selection was very effective and no escapes were observed. The study demonstrated that hygromycin resistance can be used as an effective selectable marker for plum transformation. The new system developed here is important and useful for multiple gene transformation in plum.     

Defence responses in leaves of resistant and susceptible pepper (Capsicum annuum L.) cultivars infected with different inoculum concentrations of Phytophthora capsici Leon

Three pepper cultivars (PM-702: resistant, Demre-8 and KM-hot: susceptible) with different resistances to Phytophthora capsici-22 (P. capsici-22) were inoculated with different concentrations of zoospores to analyze the time course of phenylalanine ammonia lyase (PAL) activity, malondialdehyde (MDA), hydrogen peroxide (H₂O₂), proline and total protein.     

Physiological and biochemical changes with special reference to mangiferin and oxidative enzymes level in malformation resistant and susceptible cultivars of mango (Mangifera indica L.)

Changes in biophysical attributes, mangiferin and polyphenol oxidase (PPO), catalase and peroxidase activities in malformation resistant mango cultivar Elaichi were studied at various stages of flower development and compared with susceptible cvs. Amrapali, Beauty Mc-lin and Dashehari. Accumulation of mangiferin was maximum (96.0 and 108.0 mg g⁻¹ FW) in Elaichi prior to flower bud differentiation (September) and at full bloom (February), while these were minimum (59.0 and 74.0 mg g⁻¹ FW) in susceptible cv. Beauty Mc-lin. Mangiferin promoted vegetative growth and exhibited inhibitory role on the occurrence of malformation. It was also found that the resistant cultivar had highest activity of PPO as compared to susceptible ones. There was no significant difference in the enzymes catalase and peroxidase activity at early stage of flower differentiation but at flower bud burst stage the catalase activity was enhanced significantly in cv. Elaichi (25.28 unit min⁻¹ g⁻¹ FW) in comparison to Amrapali (16.20 unit min⁻¹ g⁻¹ FW), Beauty Mc-lin (18.39 unit min⁻¹ g⁻¹ FW) and Dashehari (17.50 unit min⁻¹ g⁻¹ FW). The resistant cultivar had high leaf temperature (30.30 °C) and diffusion resistance (476.14 m mol m⁻² s⁻¹) during the flowering but the rate of transpiration and relative humidity (RH) were high in susceptible cultivars. Results of the present study clearly indicate that level of mangiferin could be considered as a potential biochemical indicator for screening mango genotypes to malformation.     

Effects of root inoculation of plant growth promoting rhizobacteria (PGPR) on yield,growth and nutrient element contents of leaves of apple

From 2002 to 2006, plant growth promoting effects of Bacillus M3, Bacillus OSU-142 and Microbacterium FS01 were tested alone or in combination on apple (Malus domestica L.) cv. Granny Smith in terms of yield, growth and nutrient composition of leaves in the province of Malatya, Turkey. The presence of M3 and/or OSU-142 and/or FS01 combinations stimulated plant growth and resulted in significant yield increases in Granny Smith. Root inoculation of PGPR strains significantly increased cumulative yield (26.0–88.0%), fruit weight (13.9–25.5%), shoot length (16.4–29.6%) and shoot diameter (15.9–18.4%) compared with the control. In addition, all nutrient element contents (N, P, K, Mg, Ca, Fe, Mn and Zn) investigated in Granny Smith, except Mg was significantly affected by bacterial applications compared with the control. The highest N content (3.38%) was obtained from OSU-142 + FS01 application. Phosphorus contents of leaves of Granny Smith increased from 0.29% in the control to 0.42, 0.37, 0.36, 0.36 and 0.35% by M3, M3 + FS01, M3 + OSU-142, OSU-142 and M3 + OSU-142 + FS01 application, respectively. All bacterial applications significantly increased Ca content of leaves. The highest K, Fe, Mn, Cu and Zn content of leaves were obtained from OSU-142, M3, FS01, M3 and M3 + FS01 applications, respectively. The results of this study suggest that Bacillus M3 and/or OSU-142 and/or Microbacterium FS01 in combination have the potential to increase the yield, growth and nutrition of apple trees.     

The conjunctive use of compost tea and inorganic fertiliser on the growth,yield and terpenoid content of Centella asiatica (L.) urban

The conjunctive use of a compost tea (CT) and an inorganic fertiliser (NPK) on the growth, yield and terpenoid content of Centella asiatica (L.) urban was evaluated. CT and NPK applied at half the recommended concentration resulted in the significant enhancement of vegetative growth, yield and antioxidant content. The synergistic effect of CT₅₀ and NPK₅₀ was highly pronounced on the bioactive components Asiatic acid, madecassoside and asiaticoside. The distribution of the total antioxidants was highest in roots (80%), followed by leaves (66%) and petioles (54%). There was a strong positive correlation (r = 0.990) between fertility and total antioxidant content, suggesting an enhanced synthesis of bioactive components resulting from the treatment. This response by C. asiatica to the integrated experimental treatment suggests a viable option for the commercial cultivation of the herb. Augmenting the vegetative biomass production and bioactive components will increase the availability of this medicinal herb for the treatment of various human ailments.     

Rose (Rosa hybrida L.) EST-derived microsatellite markers and their transferability to strawberry (Fragaria spp.)

The ‘Genome database for Rosaceae (GDR)’ provides a large collection of expressed sequence tags (ESTs) harboring simple sequence repeats (SSRs) from several Rosaceae genera, including Rosa (rose). Primer pairs flanking SSR were designed for 312 unique Rosa ESTs based on GDR database. Eight rose (Rosa hybrida L.) genotypes were tested for PCR amplification, and 287 (92%) of the primer pairs generated allele-specific PCR bands that were readily scored. From 183 (63.7%) primer pairs that evidenced polymorphic alleles among the eight rose cultivars, 20 pairs evidencing EST sequence homology to known gene functions and high levels of polymorphism were selected and utilized for DNA fingerprinting and genetic diversity assessments of 47 rose hybrids. A total of 202 polymorphic bands were scored and generated unique fingerprints for each rose hybrid. The Nei–Li genetic similarity coefficients among 1081 pair-wise comparisons of 47 cultivars exhibited a broad range of genetic variations from 0.30 (‘Grand King’ and ‘Carnival’) to 0.99 (‘First Red’ and ‘Red Champ’). UPGMA cluster analysis divided 47 hybrids into five major groups and two sub-groups. The cross-species transferability of 273 EST-SSR primer pairs was evaluated using four genotypes of the strawberry, a genus member of the Rosaceae family. PCRs on the DNA samples of strawberry were successful for 165 primer pairs; among these, 123 pairs amplified 243 polymorphic bands. As surrogates of the marker transfer, the phenetic relationship among the four strawberry genotypes was evaluated. Genetic similarity coefficients varied from 0.78 (‘Maehyang’ and ‘Janghyee’) to 0.64 (‘Janghyee’ and ‘Pragana’). The results of cluster analysis showed that the three octaploid strawberry cultivars were quite similar, whereas the diploid ‘Pragana’ was related distantly at the genomic DNA level. The EST-SSR markers developed in the present study can be efficiently utilized for genetic diversity studies in Rosaceae.     

Foraging behavior of Osmia cornifrons in an apple orchard

An apple orchard consisting of a single commercial cultivar and of pollinizers such as Crab apples under the condition of natural pollination needs pollinators for stable fruit production. We investigated the foraging behavior of pollinators, especially Osmia cornifrons Radoszkowski, in order to evaluate their use instead of honeybees.     

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.