Determination and confirmation of S-RNase genotypes of apple pollinators and cultivars

Summary

We analysed the S-RNase genotypes of 23 crab apple (Malus spp.) pollinators and 102 cultivars of domestic apple (Malus pumila Mill.) by PCR amplification and digestion. Within the 23 pollinators, four pollinators, ‘Hopa’, ‘Jack’, ‘Pink Perfection’ and ‘Profusion B’, each had two unidentified S-RNase alleles. These cultivars should be useful pollinators for all domestic cultivars. Twenty-one of the domestic cultivars exhibited S-genotypes contrary to those expected from their supposed parentage, suggesting that one or both reported parents were wrong. We confirmed many of the S-RNase genotypes by pollination tests.     

Determination of S-RNase genotypes and isolation of four novel S-RNase genes in Japanese apricot (Prunus mume Sieb. et Zucc.) native to China

Summary

Japanese apricot (Prunus mume Sieb. et Zucc.) originated in China with more than 500 cultivars. Most cultivars are self-incompatible. It is essential to identify S-genotypes for production and breeding. The S-genotypes of 17 Japanese apricot cultivars native to China were determined using the Prunus S-RNase consensus primer pair, Pru-C2 and PCE-R, which were designed from the second and third conserved regions of Prunus S-RNases, respectively. Eleven S-RNase alleles (S₁ S₂, S₇, S₁₂, S₁₄, S₁₅, S₁₈, S₂₀, S₂₂, S₂₃ and S₂₆) and four new S-RNase alleles (S₃₀, S₃₁, S₃₂ and S₃₃) with GenBank Accession Numbers JN232975, JN232976, JN232977, JN232978 were identified. Furthermore, the S-genotypes of four Japanese apricot cultivars were confirmed by field-testing for cross-pollination. The results of this study enrich the information available on S-genotypes in Japanese apricot and provide a reasonable basis for the appropriate arrangement of pollination trees in orchard practice.     

S-allele genotypes of apple pollenizers,cultivars and lineages including those resistant to scab

Summary

We examined S-allele genotypes of ten apple cultivars and species to determine their possible usefulness as pollenizers for all apple cultivars. ‘Dolgo’ did not contain any known S-RNases encoded at the S-locus, suggesting its possible usefulness as a pollenizer for almost all apple cultivars. We also identified and confirmed the S-allele genotypes of 18 apple cultivars by polymerase chain reaction (PCR)-digestion analysis. The S-genotype of ‘Kiou’ (S₁S₇), ‘Korei’ (S₃S₂₈), ‘Korin’ (S₁S₉), ‘Kotoku’ (S₁S₂₈), ‘Kyokkou (S₇S₂₅), ‘Lobo’ (S₁S₇), ’Mahe 7’ (S₂S₇), ‘Mellow’ (S₂S₃), ‘Takahara’ (S₃S₉) and ‘Warabi’ (S₉S₂₈) were confirmed by pollination results. These cultivars seemed not to have originated from the expected seed or pollen parents or, in the case of ‘Lobo’, might have been mislabelled. Finally, we identified the S-allele genotypes of ‘Prima’ (S₂S₁₀), ‘Querina’ (S₃S₉) and ‘Yoko’ × ‘Prima’ (S₃S₁₀), which are resistant to scab.     

Identification of S-haplotypes of European cultivars of sour cherry

The sour cherry is known to exhibit the phenomenon of gametophytic self-incompatibility which prevents self-fertilization. In sour cherry, besides self-incompatible cultivars, there often occur self-compatible cultivars. This is due to the occurrence of natural mutations of the S-RNase or SFB genes and, consequently, loss of functionality of S-alleles. Here we present the results of the identification of S-haplotypes of 21 cultivars of sour cherry from various regions of Europe. The analyses were performed using methods based on the amplification of intron I and intron II of the S-RNase gene and fragments specific to the individual alleles of the S-RNase or SFB genes. The tested cultivars were found to contain 15 S-haplotypes: S₁, S_1?, S₄, S₆, S_6m, S_6m2, S₉, S₁₂, S₁₃, S_13?, S₂₆, S₃₅, S_36a, S_36b, and S_36b2. The most frequently occurring S-haplotypes were S_13? (61.9%), S_36a (57.1%), and S₂₆ (47.6%). On the basis of the results, 17 of the 21 cultivars were deduced to be self-compatible. The results will be of use in the production of sour cherry fruit by facilitating the selection of suitable pollinating cultivars. The results are also expected to be useful in the breeding of new cultivars of sour cherry when selecting genotypes for crosses.     

Physiological Breakdown in Stored Monarch Plums

Summary

Several pollination studies carried out on different, self-incompatible almond cultivars and seedlings have shown the presence of variable levels of fruit set following self-pollination that could be attributed to partial self-incompatibility (PSI). PSI is an intermediate reproductive behaviour, described in some angiosperm species, which indicates that self-incompatibility is a quantitative and plastic trait. The present study was performed to substantiate the occurrence of PSI in almond by studying ten cultivars that are traditionally considered to be self-incompatible (four of which had previously shown fruit set after self-pollination). These cultivars were analysed by microscopic observations of the progression of pollen tubes through the pistil following controlled self-pollination, determinations of fruit set after bagging flower buds, and molecular identification of the parentage of the fruits obtained using consensus PCR-primers for Prunus S-RNase alleles. The results showed that, in nearly all cases, the pollen tubes did not enter the ovary and, from a total of 5,349 bagged flower buds, only 17 fruits were obtained. In all cases, PCR analysis of the plants obtained after germination of these seeds showed a band corresponding to an S-RNase allele not present in the maternal progenitor, clearly indicating that they could only have arisen from cross-pollination.Therefore, the low fruit set values observed in some of the cultivars studied were not due to a breakdown in the self-incompatibility response that confers PSI, but to very low rates of contamination with foreign pollen. These results corroborate the self-incompatibility phenotype of the cultivars studied here, and highlight the importance of ascertaining the identity of the parentage in any progeny obtained after bagging.     

Determination of self-incompatible genotypes in 21 cultivated sweet cherry cultivars in Greece and implications for orchard cultivation

Summary

Sweet cherry is self-incompatible due to having a gametophytic self-incompatibility system. S alleles in the style and pollen determine possible crossing relationships. Knowledge of the S allele constitution of cultivars is important for sweet cherry growers and breeders. Recently, molecular methods have been developed to distinguish between S alleles in sweet cherry.The S allele genotypes of 21 sweet cherry cultivars widely grown in Greece, including 19 not previously genotyped, were determined based on their S-RNase gene sequences using PCR analysis. Eight different S alleles in ten combinations were distinguished and two new S-genotypes (S₁S₁₃ and S₄S₃₀) were documented. Four alleles, S₁, S₃, S₄, and S₉ were widespread and together were responsible for 85% of the S-haplotypes. Therefore many of the cultivar combinations were semi-compatible. In Greece, semi-compatibility was shown to correlate with low yields. However, the cultivar ‘Hybrid Tragana Edessis x Unknown’ (S₃S₁₃) and the cultivar ‘Kapsiotika’ (S₂S₅) carry rare S-haplotypes and are therefore fully cross-compatible with most of the cultivars analysed.     

Agronomic evaluation of seedlings from crosses between the main Spanish olive cultivar ‘Picual’ and two wild olive trees

Summary

Until recently, olive breeding programmes have been based exclusively on cultivated olive germplasm (Olea europaea subsp. europaea var. europaea), in contrast to breeding in other fruit crops where the use of wild relatives has been widely reported. In this study, ten agronomic traits were evaluated in two progenies derived from controlled crosses between the Spanish olive cultivar ‘Picual’ (female) and two wild (O. europaea subsp. europaea var. sylvestris) genotypes (males). The results of this evaluation were compared with the progenies of crosses between ‘Picual’ (female) and the three cultivars, ‘Sikitita’, ‘Meski’, and ‘Zaity’ (male). The two ‘Picual’ × wild genotype progenies had the highest mean values for vigour traits (i.e., tree height and trunk diameter). Progenies from both ‘Picual’ × wild genotype crossess also showed the highest percentages of trees (53.8% and 37.5%) with a short juvenile period, compared to the progenies from crosses between ‘Picual’ and each of the three cultivars ‘Sikitita’, ‘Meski’, and ‘Zaity’ (0%, 5%, and 3.6%, respectively). Significant differences were also observed between progenies in fruit traits as well as in oil contents and fruit weights. Progenies from both crosses with wild olive showed lower values for the fruit traits evaluated than the cultivated olive progenies. However, significant improvements were achieved compared to fruit traits in the wild parents. The implications of these results for the future use of wild germplasm in olive breeding programmes are discussed.     

Peroxidase isoenzyme genes in the identification of apple cultivars and Malus species

The peroxidase genotypes of 188 apple cultivars, rootstocks and Malus species were recorded. Four loci, PRX-2, PRX-3, PRX-4 and PRX-7 were involved including 15 alleles. Using this information a clear grouping of 83 cultivars, 46 rootstocks and 59 species was achieved through knowledge of the genetic basic of banding patterns, which consequently enabled secondary and inconsistent bands to be recognised and disregarded as relevant factors in identification. The possible selective advantage of the PRX-2e allele recognised in descendants of ‘Jonathan’ (PRX-2ce) is discussed. Triploid cultivars showed three different PRX-2 alleles including ‘Ashmead’s Kernel’ (PRX-2bce), providing further evidence of the triploid nature of this cultivar.     

Creating triploid germplasm via induced 2n pollen in Capsicum annuum L.

Summary

2n pollen of diploid chili (Capsicum annuum L. ‘Xianjiao’) was induced by heat shock (35.5?C, 4.h) or low temperature treatment (118C, 48.h). The percentages of 2n pollen were increased to 8.3% and 10.5% respectively. The cytogenetic mechanisms of 2n pollen formation were first division restitution, second division restitution and abnormal cytokinesis. Diploid 2n pollen was less competent than n pollen in fertilization. Only one triploid plant arose from 78 progenies after fertilization by 2n pollen. The four offspring plants of the triploid were diploid. The morphological characters of the triploid and microsporogenesis were also evaluated.     

Evaluation of an alternative D-amino acid/DAAO selection system for transformation in apple (Malus × domestica Borkh.)

Summary

The conventional selection system for apple transformation is based on the selectable marker gene, nptII, encoding antibiotic resistance against kanamycin. We tested an alternative selection system based on the use of D-amino acids using the gene, D-amino acid oxidase 1 (dao1) as the selectable marker, in order to avoid the presence of antibiotic resistance genes in the resulting transgenic apple plants. In addition, dao1 allowed the selection as well as the elimination of dao1-transgenic plants, based on differences in the toxicity of different D-amino acids. Regeneration experiments using apple leaf explants revealed that 2 mM D-serine or D-alanine inhibited shoot regeneration. We performed transformation experiments using the apple cultivars ‘Gala’, ‘Holsteiner Cox’, and a progeny of the apple cultivar ‘Pinova’, and the vector p35S::dao1-intron, containing the dao1 and nptII selectable marker genes. Several shoots regenerated successfully on selection media containing various concentrations of D-serine or D-alanine, but transgenic shoots were not obtained. However, three dao1/nptII transgenic apple lines were obtained after selection with kanamycin, indicating that the vector was functional. Furthermore, we showed that 20 mM D-serine could be used to select dao1-transgenic shoots from non-transgenic in vitro shoots, whereas 13 mM D-isoleucine had the opposite effect.     

Characterization of interspecific hybrids of Petunia and Calibrachoa by multiplex PCR,DNA content,and chromosome number

Petunia and Calibrachoa are ornamental plants of worldwide economic importance. Commercially, it is useful to combine desirable characteristics of Calibrachoa, including the large diversity of flower colors, with the pH tolerance and vigorous growth of Petunia. For investigation of interspecific hybrids between Petunia and Calibrachoa we used four different introns with flanking exon areas, each from a different gene and with a different sequence for Petunia and Calibrachoa, as genus-specific markers for chromosomes in interspecific hybrids. PCR primers specific for Petunia and Calabrachoa were made from the determined sequences and tested on a range of presumed hybrids using multiplex PCR, which confirmed that the hybrids were derived from these two species. Flow cytometry demonstrated that the hybrids were triploid, and determination of chromosome numbers showed that two of the chromosome sets were from Calibrachoa, i.e. that the other parent, Petunia, was diploid. Similar methods can be used for molecular characterization of other new hybrids made between closely related species or cultivars.     

Multiple Avenues to Gender in Strawberries

ABSTRACT

Common strawberry cultivars are hermaphrodites, producing both anthers and pistils in their flowers. However, octoploid Fragaria species are trioecious and different genotypes can be female with pistillate flowers, hermaphrodites, or males with staminate flowers. One female selection, F. virginiana High Falls 22 and three hermaphrodite selections, N8688, RH23, and Montreal River 10 were hybridized with five June-bearing cultivars. Seedlings of each family were planted in Ontario and Michigan. Two F. chiloensis selections Pigeon Point (female) and FRA1267 (hermaphrodite), were hybridized with 14 F. virginiana selections. Seedlings from each family were planted in Ontario and Minnesota. In all progeny, the presence or absence of anthers was recorded and a subjective scale used to estimate the percentage of fruit set (1–10 representing 10% intervals). Also, FRA1267 was crossed with one F. × ananassa selection and the progeny intercrossed and grown in Ontario. In the sib-crosses gender was recorded. In two female parents, the female progeny had on average a higher fruit set than the hermaphrodite progeny. F. virginiana genotypes varied in their fruit set, which suggested that there are a number of alleles involved in the fertility of hermaphrodites. In crosses, FRA1267 produced 80% females when used as a female parent and 100% hermaphrodites when used as a male parent. It is thought that the trait is inherited cytoplasmically or that the F. chiloensis alleles involved are epistatic when F. virginiana or F. × ananassa is used as a male parent.     

Cloning of the S25 cDNA from ‘McIntosh’ apple and an S25-allele identification method

Summary

An S₂₅-RNase gene in the pistil responsible for self-incompatibility within the apple (Malus × domestica Borkh.) was cloned from a McIntosh cultivar. We developed an S₂₅-allele-specific polymerase chain reaction and digestion (PCR-digestion) method using specific primers and the restriction enzyme BamHI. We investigated the S-allele genotypes of 18 ‘McIntosh’ progeny.     

Production and characterization of new triploid seedless progenies for mandarin improvement

Eighty-six triploid Citrus plants were recovered from interploid crosses between a natural tetraploid selection of the tangerine ‘Dancy’ (Citrus reticulata Blanco, cultigroup ‘Tangerine’), used as the pollen parent, and two seedy selections of diploid mandarins (C. reticulata Blanco) as well as one clementine (Citrus clementina Hort. ex Tan.): ‘Fortune’ mandarin, ‘Wilking’ mandarin and ‘Monreal’ clementine. Flow cytometric analysis was used for screening the triploid plantlets and the ISSR-PCR technique was used to characterize the obtained triploids through a double approach, confirming the hybrid nature of the offspring and allowing the analysis of the genetic pool obtained. Selection among triploid genotypes generated from elite seedy parents will give us a better chance to obtain superior mandarin cultivars characterized by true seedlessness.     

Effects of grafting height of MM106 rootstock on growth,lateral shoot formation and yield in apple trees

Summary

Between 2004 and 2008, the effects of different grafting heights on sylleptic shoots were tested in the apple (Malus domestica Borkh.) cultivars ‘Granny Smith’ and ‘Gloster’, and cumulative fruit yields were evaluated. MM106 apple rootstocks were grafted 10, 20, 40, or 60 cm above soil level in August 2004. The results showed that an increased grafting height significantly decreased tree height in both cultivars. The tallest and shortest trees were observed at grafting heights of 10 cm (153.0 and 170.0 cm) and 60 cm (141.3 and 143.5 cm) in ‘Granny Smith’ and ‘Gloster’, respectively.Among the various grafting heights tested, 60 cm in ‘Granny Smith’ and 20 cm in ‘Gloster’ gave the largest stem diameters (17.6 mm and 16.8 mm, respectively). The number of lateral shoots increased significantly with increased grafting height in both cultivars. The largest numbers of lateral shoots in ‘Granny Smith’ (10.75) and ‘Gloster’ (2.00) were obtained from a grafting height of 60 cm, while 2.55 and zero lateral shoots occurred at 10 cm grafting height in ‘Granny Smith’ and ‘Gloster’, respectively. Shoot lengths decreased significantly by increasing the grafting height. Grafting heights of 10 cm and 60 cm resulted in the tallest and shortest shoots in both cultivars. Cumulative fruit yields were significantly affected by grafting height in both cultivars. The highest yield was found for a 60 cm grafting height in both ‘Granny Smith’ (11.295 kg tree^–1) and ‘Gloster’ (4.818 kg tree^–1).The results of this study suggest that grafting heights of 40 cm and 60 cm have the potential to promote branching and early bearing for apple fruit production in sustainable and organic agricultural systems.     

A novel method for Citrus propagation: Seed grafting

Summary

We have developed a novel technique for grafting citrus seeds onto citrus rootstock plants that resulted in successful graft-take with normal vascular connections between the emerging seedling stem tissues and the rootstock plant. The method was found to be suitable for producing grafted plants from seeds of six cultivars and hybrids of Citrus and the citrus relative Murraya paniculata, using four common Citrus rootstocks. Plants produced by this method developed normally and were established in the field more rapidly than those produced by the common practice of grafting the rootstocks with budwood derived from seedlings prepared from seed in soil-based media. Seed grafting is expected to find a range of uses in breeding programmes; for example, by reducing the time required for the evaluation of hybrid seedlings, in cases where the female parent is mono-embryonic, for testing for vertical transmission of pathogens, and for screening for pathogen resistance among hybrid and mutagenised seed sources.     

Ultrastructure of epicuticular wax aggregates during fruit development in apple (Malus domestica Borkh.)

Summary

Development of wax platelets on the surface of ‘Delicious’ (Malus domestica Borkh.) apple fruit was investigated throughout the growing season using field emission scanning electron microscopy. At 5,000 and greater, wax crystalline structures appeared to be composed of microtubules (MT), aggregates of individual MT to form single platelets, and of one or more platelets. The thickness of a single wax platelet ranged from 116–128 nm, approximately the diameter of a single MT; whereas multiple-platelet aggregates ranged in thickness from 307–428 nm. Individual platelets, multiple platelet aggregates and MT were visible throughout fruit development, as well as on different apple cultivars. Examination of the cuticle from young fruitlets (receptacle diameter = 3 mm) of ‘Chinese Crabapple’ (M. hupehensis Rehd.), which develops without trichomes, best demonstrated early platelet formation. A model for wax platelet and cuticle development on apple is proposed based on these data.     

Breeding Strawberry for Higher Phytochemicals Content and Claim It: Is It Possible?

ABSTRACT

Commercial strawberries (Fragaria × ananassa) are one of the most investigated berries for nutritional and nutraceutical properties. A strawberry breeding program, including inter-specific back-crosses, is now producing new genotypes with increased vitamin C, folates, and phenols combined with important commercial traits. The release of new cultivars with a high content of bioactive compounds, which may contribute to health, wellness, and reduced disease risk, is expected from this program. The ultimate goal is to properly obtain and integrate new scientific knowledge from genetic, agronomic, and biomedical studies that may bring acceptance from food agencies for specific health claims on new strawberry cultivars.