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.     

Identification of seven haplotype-specific SFBs in European pear (Pyrus communis) and their use as molecular markers

The European pear (Pyrus communis) carries the S-RNase-mediated gametophytic self-incompatibility (GSI) system. The S-haplotype is conferred by an S-locus, which contains the style-specific expressed S-RNase, and the pollen-specific expressed F-box genes (SFB). Both the S-RNase and the SFB genes are multi-allelic and each is characteristic of one of the S-haplotypes. Therefore, they are ideal markers for molecular S-genotyping. In this work, for the first time, seven haplotype-specific SFBs were isolated from European pears. Particular primers for each of these SFBs were generated, thus providing an additional tool for S-genotyping of European pear cultivars.     

Performance of ‘Coscia’ pear (Pyrus communis) on nine rootstocks in the north of Israel

The vegetative and reproductive performances of ‘Coscia’ pear (Pyrus communis L.) grown on nine rootstocks [OHF 69, OHF 97, OHF 217, OHF 333, OHF 513 and BP 1 (P. communis), clonal seedling (Davis AxB) of Pyrus betulifolia and quince BA 29 and EMA (Cydonia oblonga)] were compared during a 9-year period. The trial was conducted at the Experimental Station ‘Avnei Eitan’ in northern Israel (elevation 400 m above sea level, a.s.l.), on a well-drained soil with pH 7.1. Trees were planted in December 1998, spaced at 4.0 m × 2.0 m and trained with a central axis. The most vigorous trees were on P. betulifolia seedlings, followed by the four OHF rootstock (69, 97, 333, 513) and BP 1 (with no significant difference between them). All the above rootstocks demonstrated greater vigor than OHF 217 and quince BA 29 or EMA. The highest cumulative yields per tree were harvested from trees on the four OHF rootstocks (69, 97, 333, 513) and P. betulifolia followed by the BP 1. The two quince rootstocks, as well as OHF 217, had the lowest cumulative yield and the lowest yield of large fruit. A positive correlation was found between the vigor of the tree, as affected by the rootstock, and both the total yield and the fruit size. We conclude that in a warm climate, yield efficiency is not the only parameter that should be taken into account, and building a strong tree for a weak cultivar is the first requirement for establishing an orchard.     

Molecular and physiological identification of new S-alleles associated with self-(in)compatibility in local Spanish almond cultivars

The S-allele characterisation of ‘Alzina’ and ‘Garondès’, two local almond cultivars from the island of Majorca, by a multidimensional approach has allowed the confirmation of the presence of the S_f-allele and the identification of a new allele not previously described in almond, S₃₆. When these cultivars were phenotypically evaluated, both showed a self-incompatible phenotype and were cross-incompatible, as assessed by artificial self- and cross-pollinations and fruit sets after field pollinations, confirming that their S_f-allele is in its active form, S_fa. Thus a new CGI group in almond is proposed and named XXVIII. These results confirm the wide diversity of S-alleles in almond both at genotypic and phenotypic levels, as well as their similarity with the S-alleles from other close Prunus species. This similarity suggests the possibility of allele introgression between species or allele identity by descent from a common ancestor.     

Genetic diversity in local Tunisian pears (Pyrus communis L.) studied with SSR markers

Few records are available about local Tunisian pear cultivars characterized by low chilling requirements and adaptation to dry conditions. In this work, seven SSRs derived from apple were successfully transferred to 25 local Tunisian pear genotypes and 6 common varieties of Pyrus communis cultivated in Europe. The 7 SSRs used amplified a total of 36 fragments. All the microsatellites except one seem to amplify more than one locus in some of the genotypes studied. Only 12 different fingerprinting patterns could be distinguished among the 25 Tunisian cultivars studied indicating a high number of synonymies. The mean expected and observed heterozygosities in the 25 Tunisian cultivars analyzed averaged 0.71 indicating a high level of genetic diversity among the local Tunisian pear germplasm. These markers will be useful to optimize the conservation of this highly threatened germplasm.     

S-RNase based S-genotyping of Japanese plum (Prunus salicina Lindl.) and its implication on the assortment of cultivar-couples in the orchard

Determination of the genetic compatibility between self-incompatible cultivars is crucial in agriculture. The Rosaceae family carries the S-RNase-mediated gametophytic self-incompatibility (GSI) system. Each haplotype is conferred by an S-locus. The S-locus contains two highly polymorphic genes, S-RNase and SFB, which are characteristic of each haplotype and therefore these genes are ideal markers for molecular S-genotyping. In this study 43 Japanese plum cultivars grown in Israel were S-genotyped based on their S-RNase gene sequences. Four alleles, S^b, S^c, S^e and S^h are widespread and together are responsible for 87% of the S-haplotypes therefore many of the cultivar combinations are semi-compatible. In Israel semi-compatibility was shown to correlate with low yield. However, two cultivars, ‘Wickson’ S^fS^k and ‘Shiro’ S^fS^g carry rare S-haplotypes and, therefore, are fully compatible with most of the analyzed cultivars.     

Identification of S-genotypes and novel S-RNase alleles in Japanese apricot cultivars native to China

Information on S-genotypes is essential for designing orchards of Japanese apricot (Prunus mume Sieb. et Zucc.) and for hybridization. However, this information is lacking for most cultivars grown in China. Thus, in this work, the S-genotypes of 24 Japanese apricot cultivars native to China were identified by sequencing the PCR products obtained from allele-specific polymerase chain reaction (AS-PCR). Seventeen S-RNase alleles were amplified, ten of them for the first time. The new S-RNase alleles were submitted to GenBank and denoted them as S¹⁷, S¹⁸, S¹⁹, S²⁰, S²¹, S²², S²³, S²⁴, S²⁵ and S²⁶. Furthermore, the S-genotypes of four Japanese apricot cultivars were confirmed by field-testing cross-pollination.     

Nitrogen distribution,remobilization and re-cycling in young orchard of non-bearing ‘Rocha’ pear trees

In newly planted orchards, special attention must be paid to fertilization to build up the permanent structure of the trees so that high yield and fruit quality can be reached later on. Nitrogen (N) plays a major role in the fertilization plan, although few studies have assessed its use efficiency in young non-bearing trees, especially in field conditions. In this work, 1–3 years old ‘Rocha’ pear trees, grafted on quince BA29, were planted in a Mediterranean region, and fertigated with 6 g N tree⁻¹ year⁻¹ as ammonium nitrate with 5 at.% ¹⁵N enrichment to study the fertilizer N uptake during the vegetative cycle, the overall fertilizer N use efficiency at the end of each year, and the plant–soil N balance for this period. Nitrogen remobilization and the re-cycling of N from senescent leaves were also studied by fertilizing some pear trees with 10 at.% ¹⁵N enrichment.     

Relationship between inbreeding coefficients and plant height of 1-year-old seedlings in crosses among Japanese pear (Pyrus pyrifolia Nakai) cultivars/selections

Plant height, a vigor trait, in 1-year-old seedlings made from Japanese pear (Pyrus pyrifolia) cultivars/selections was measured using 994 individuals from 29 families. The family mean of plant height was negatively correlated (r = −0.72**) to the inbreeding coefficients (F). The regression of the family mean (Fm) on the F value (Fm = 130 − 104F) showed that inbreeding depressions were 8%, 20%, and 40% for F = 0.1, 0.25, and 0.5, respectively. According to the regression, the family mean at F = 0 was estimated at 130 cm. These results showed that the vigor was greatly influenced by inbreeding in Japanese pear. Within-family variances, the genetic segregation of offspring in a family, differed according to family. The proportions of offspring with plant height above 130 cm (estimated Fm for F = 0) were extremely low, i.e., 0–17% for 0.5 ≤ F < 0.60 and 0–8% for F = 0.75.     

Content levels of various fruit metabolites in the ‘Conference’ pear response to branch bending

The response of ‘Conference’ pear subjected to branch bending in the content levels of various metabolites in its fruit was investigated. The fruits in commercial maturity were sampled in 2004 and 2005 from branches bent in the late summer of 2003 (the summer treatment), from branches bent in the late spring of 2004 (the spring treatment) and from control branches. The content levels of carbohydrates, organic acids and phenolic compounds were compared among treatments in two successive years. The fruit revealed various responses in content levels of metabolites. In the first year after bending, no significant differences were found in the content of each carbohydrate, but in 2005 by far the highest content level of glucose and fructose and the lowest content level of sucrose were found in fruits from the control. The control fruit showed significantly higher content levels of malic acid and lower content levels of some phenolics (chlorogenic and vanillic acid) in the first year after bending, but in the next year the opposite reaction occurred—the control fruit had the lowest content level of malic acid and the highest content level of epicatechin, quercetin-3-d-galactoside and quercetin-3-β-d-glucoside. The comparison of the two bending treatments alone in 2004 showed that the summer treatment often produced a slightly higher value of each phenolic in comparison to the spring treatment. However, in 2005 the significantly highest content of chlorogenic acid was in fruit from the spring treatment. Sorbitol, as well as citric acid, catechin and sinapic acid showed no clear tendency among treatments, neither in 2004 nor in 2005. It is suggested that these variations of ‘Conference’ fruit subjected to different bending treatments could not be the result of bending alone, but that they could be indirectly affected by other physiological responses of the fruit tree. However, it seemed that variations are affected by the time of bending and by the year-to-year, and such responses can be attributed to the ‘Conference’ genotype only.     

Fruit thinning in ‘Conference’ pear grown under deficit irrigation: Implications for fruit quality at harvest and after cold storage

Fruit thinning in pear is feasible for mitigation of water stress effects. However, it is not well known how fruit quality at harvest and after cold storage is affected by pre-harvest water stress. Even less is known about the effects of fruit thinning on quality under these circumstances. To elucidate these, we applied deficit irrigation (DI) and fruit thinning treatments to ‘Conference’ pear over the growing seasons of 2008 and 2009. At the onset of Stage II (80 and 67 days before harvest in 2008 and 2009, respectively), two irrigation treatments were applied: full irrigation (FI) and DI. FI trees received 100% of crop evapotranspiration (ETc). DI trees received no irrigation during the first three weeks of Stage II to induce water stress, but then received 20% of ETc to ensure tree survival. From bud-break until the onset of Stage II and during post-harvest, FI and DI trees received 100% of ETc. Each irrigation treatment received two thinning levels: no thinning leaving commercial crop load (∼180 fruits tree⁻¹), and hand-thinning at the onset of Stage II leaving a light crop load (∼85 fruits tree⁻¹). Under commercial crop loads, DI trees were moderately water-stressed and this had some positive effects on fruit quality. DI increased fruit firmness (FF), soluble solids concentrations (SSC) and acidity at harvest while no changes were observed in fruit maturity (based on ethylene production). Differences in FF and acidity at harvest between FI and DI fruit were maintained during cold storage. DI also reduced fruit weight loss during storage. But fruit size was reduced under DI. Fruit thinning under DI resulted in better fruit composition with no detrimental effect on fresh-market yield compared to un-thinned fruit. Fruit size at harvest and SSC values after five months of cold storage were higher in fruit from thinned trees than fruit from un-thinned trees. Fruit thinning increased fruit ethylene production, indicating advanced maturity. This may lead to earlier harvest which is desirable in years with impending drought. Fruit thinning is therefore a useful technique to enhance pear marketability under water shortage.     

Characterization of variability and genetic similarity of European pear using microsatellite loci developed in apple

Seven microsatellite loci (SSR) developed in apple were used for the identification of 63 European pear cultivars. A total of 46 fragments were amplified, with an average of 6.6 alleles per SSR. Only one microsatellite amplified more than one locus. The mean expected and observed heterozygosities over the six single-locus SSRs averaged 0.68 and 0.44, respectively, and the number of effective alleles per loci was 3.43. The amplified fragments produced 61 different fingerprinting patterns that allowed to unequivocally distinguish all the varieties analyzed. Only two varieties, which derive from mutations, could not be distinguished from the original variety. Cluster analysis of the estimated genetic similarity, grouped the varieties according to their pedigree and their geographic origin. The variability detected with the SSRs in European pear varieties was low when compared with the variability detected in other fruit crops in the Rosaceae.     

Identification of parental species of the Alstroemeria cv. ‘Jubilee’ using AFLP marker technique

Twelve Alstroemeria species, two hybrids, one cv. ‘Jubilee’, an anther-cultured plant from cultivar ‘Jubilee,’ and Bomarea salsilla and Leontochir ovallei (the latter two were chosen as outgroup) were evaluated using the AFLP marker technique in order to identify putative parental genotypes of the Alstroemeria cv. ‘Jubilee’ and of known interspecific hybrids, and to estimate their genetic relationships within the genus Alstroemeria. A total of 297 AFLP markers were scored by using the primer combination (E + ACCA/M + CTAG). In order to discriminate all Alstroemeria genotypes, cluster analysis (UPGMA) and principal coordinates analysis were performed. The Alstroemeria cv. ‘Jubilee’, of which the parents are unknown, had genetic distance (GD) 0.54 from Alstroemeria exserens, GD 0.57 from Alstroemeria garaventae, GD 0.62 from Alstroemeria gayana, and GD 0.66 from Alstroemeria hookeri cumminghiana. Thus, these four species are considered as putative parental genotypes. An interspecific hybrid (Alstroemeria aurea × Alstroemeria inodora), showed the smallest genetic distance from A. aurea (GD 0.56) and A. inodora (GD 0.45). The Alstroemeria ligtu group was distantly allocated from other Chilean species. We conclude that the AFLP marker technique appears to be a satisfactory tool for identifying the parental genotypes of interspecific hybrids in Alstroemeria.     

Feathering of ‘Doyenné du Comice’ pear in the nursery using repeat sprays of benzyladenine and gibberellins

This work sets out to examine the effectiveness of a number of cytokinins or products with cytokinin-like activity, with and without additional sprays of gibberellins or leaf plucking, on the production of feathers (sylleptic shoots) on trees of the European pear cultivar ‘Comice’ in a fruit tree nursery. It covers the results from three successive years experimentation.     

Effect of Glomus mosseae and Entrophospora colombiana on plant growth,production, and fruit quality of ‘Maradol’ papaya (Carica papaya L.)

The effect of inoculating ‘Maradol’ papaya plants with arbuscular mycorrhizal fungi (AMF) Glomus mosseae (GM) and Entrophospora colombiana (EC) was assessed. The results showed that both mycorrhizae species increased the number of fruits and yield in papaya plants by 41.9 and 105.2% for GM and 22.1 and 44.1% for EC, respectively, with respect to control plants. GM significantly increased plant height. Sugar content, firmness, color (°Hue), and ripening process of mycorrhized plant fruits were similar to those of the control. Weight loss of mycorrhized plant fruits was considerably less than that of the control. Inoculation of papaya with AMF is recommended, particularly with GM since it increases yield, and fruit weight (45.1%), furthermore, it reduced fruit weight loss during ripening.     

Agrobacterium × plant factors influencing transformation of ‘Joseph's coat’ (Amaranthus tricolor L.)

A protocol is developed for Agrobacterium-mediated genetic transformation of Amaranthus tricolor via explant co-cultivation with Agrobacterium rhizogenes. Bacteria-plant specific factors which influenced transformation were optimized. Of the two Agrobacterium strains employed, LBA9402 was more infectious compared to A4. Bacterial suspensions grown overnight with 100 μM acetosyringone and experiencing O.D.₆₆₀ = 0.6 followed by dilution to a density of 10⁹ cells ml⁻¹ were the most effective. Explants from garden-grown plants were more responsive than those from in vitro cultures; stem internodes being better than leaves. Immersion of the pre-pricked explants in bacterial suspension resulted in a markedly higher transformation frequency compared to the direct injection method. The infection of internode explants with the LBA9402 strain followed by co-cultivation on growth regulator-free MS medium (MS0) for 5 days resulted in emergence of hairy roots up to a maximum frequency of 97.22%. Roots were individually cultured in MS0, but fortified with bactericidal antibiotic (500 μg ml⁻¹ cefotaxime). Rhizoclones showing prolific growth were renewed through successive subcultures in MS0. Opine gene expression was revealed by positive agropine and mannopine synthesis in all selected transformed rhizoclones. Shoot regeneration from root clones, capable of auxin-independent growth and opine proficiency, was stimulated in MS augmented with 2.0 mg l⁻¹ zeatin. pRi TL–DNA rolB and pRi TR–DNA man2 ORF were detected in leaf tissues of regenerated plants from selected hairy root clones through PCR amplification. The implication of such findings is discussed on the possibility of conferring protection to crop amaranths against biotic stress challenges, particularly due to insects, viruses or fungal pathogens.     

H2O2 metabolism during sweet orange (Citrus sinensis L. Osb.) ‘Hamlin’ Xanthomonas axonopodis pv. citri interaction

Sweet orange (Citrus sinensis L. Osb.) ‘Hamlin’ is a canker (Xanthomonas axonopodis pv. citri: Xac) susceptible citrus genotype grown commercially worldwide. Canker causes severe economic losses and restricts the marketability of crop for export. Little is known about the role of oxidative stress in canker development. In the present investigation, sweet orange ‘Hamlin’ leaves were artificially inoculated with Xac to determine the impact of Xac infection on hydrogen peroxide (H₂O₂) metabolism. Characteristic symptoms following artificial inoculation were water soaking of the infiltrated zone between 2 and 8 days after inoculation (dai); raised epidermis accompanying tiny yellow colored bacterial colonies at 8 dai; and yellowing and necrosis of the infected zone by 12–16 dai. In planta Xac population increased 1000 fold by 14 dai from an initial population of 7.3 × 10⁶ cfu cm⁻² (0 dai). Peak concentrations of H₂O₂ were observed at 24 h and between 8 and 10 dai and coincided with higher activity of total superoxide dismutase (SOD). Lower levels of H₂O₂ in infected leaves were maintained by Xac induced higher activities of catalase (CAT), ascorbate peroxidase (APOD), and guaiacol peroxidase (POD). It appears Xac altered H₂O₂ metabolism in C. sinensis L. Osb. ‘Hamlin’ to enhance survival and growth.     

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.     

Evaluation of ‘Tonda di Giffoni’ hazelnut (Corylus avellana L.) clones

‘Tonda di Giffoni’ is among the most highly appreciated Italian hazelnut (Corylus avellana L.) cultivars. Due to its round kernels and excellent processing quality, it was awarded a Protected Geographical Indication (PGI) from the European Union. To identify clones expressing improved nut and production qualities, a ‘Tonda di Giffoni’ clonal selection programme was conducted across hazelnut orchards in the Irno valley of Italy from 1995 to 2006. One hundred different clones were selected and propagated in a replicated trial under similar climate, soil, and cultural conditions. From this work, the 29 best clones were identified and from 2006 to 2008 their agronomic and pomological characteristics were observed. Microsatellite or simple sequence repeat (SSR) markers were used to successfully confirm true-to-type identity of the clones. Traits evaluated included flowering time (anthesis), bud break, suckering, trunk diameter, nut and kernel characteristics and productivity (yield). Best linear unbiased predictions for clone means and estimates of intraclass correlation coefficient were obtained using R environment, lme4 and ggplot2 packages. Five clones superior to that of the standard of ‘Tonda di Giffoni’ were identified in this study. Furthermore, yield and number of suckers produced showed sufficient variability to likely be exploited for breeding. The selected clones express features useful for both growers and the processing industry and will be propagated and planted in hazelnut orchards for further study and commercial production.