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.     

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 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.     

Phenolic Compounds,Antioxidant Activity,Fatty Acids and Volatile Profiles of 18 Different Walnut (Juglans regia L.) Cultivars and Genotypes

The study was carried out to determine phenolic compounds, antioxidant activity, total lipids, saturated fatty acids, unsaturated fatty acids, and volatile organic compounds (VOCs) of 18 different walnut (Juglans regia L.) cultivars and genotypes grown in Usak province, Turkey. The biochemical compounds were identified for each cultivar and genotype and a comparative evaluation was carried out. The results showed that most of the biochemical compounds are significantly varying among each other (p?<?0.05). The existence and abundance of the biochemical compounds in the cultivars and genotypes have not only found to be a cause of differentiation, but they have been important parameters for the similarities among the cultivars and genotypes. Total phenolic content and antioxidant activity ranged from 750.67 to 1245.64?mg GAE ml^?1 and from 42.46 to 56.50% in kernels, respectively. The highest contents of the phenolic compounds were noted from gallic acid (11991?mg kg^?1), ellagic acid (1057.16?mg kg^?1), catechin (425.4?mg kg^?1), and rutin (216.6?mg kg^?1), as an average of all cultivars and genotypes. The highest lipid contents were noted from ‘Franquette’ and ‘Fernette’ as 64.28% and 63.26%, respectively, while the lowest content was noted from ‘Oguzlar 77’ (52.52%). A total of 42 VOCs were described from the 18 different walnut cultivars and genotypes. Overall, results showed that the phenolic profiles, fatty acids composition, and VOCs play an important role in similarities and diversities among the cultivars. According to the results, 7 different clusters were developed from the 18 cultivars and genotypes. Herein, ‘Fernor’, ‘Maras 18’, ‘Arslan Local Type’ and ‘Cisco’ cultivars were found to have the lowest biochemical compositions. The superior cultivars or genotypes, in terms of the abundance of the biochemical composition, were found to be ‘Gulizar Hanim Type’, ‘Lara’, ‘Pedro’, ‘Tulane’, ‘Fernette’, ‘Balkan’ and ‘Franquetta’, where they were found to develop 3 clusters by 1:3:3, respectively.

     

Determining the S-genotypes of several sweet cherry cultivars based on PCR-RFLP analysis

Summary

Based on the cDNA sequences encoding sweet cherry self-incompatibility associated ribonucleases (S-RNases), a PCR-based S-allele typing system for sweet cherry cultivars has been recently developed. Using this technique, we determined S-genotypes of the three newly released Japanese cvs Kouka-Nishiki, Beni-Sayaka and Beni-Shuho and one British cv Merton Glory that was classified as a Universal Donor, which is able to be used as a pollen donor for all cultivars in pollen incompatibility groups I to XIII. Furthermore, we also determined the partial sequences of the S-RNase genes of ‘Rainier’ (S¹S⁴)‘ and ‘Sato-Nishiki (S³S⁶)’,which leads to the development of a more reliable S-allele identification method of PCR-RFLP for sweet cherry cultivars. Total DNA isolated from leaves of the four cultivars along with those from ten cultivars with known S-genotypes were PCR amplified with two sets of primers that were designed from DNA sequences encoding the signal peptide (Pru-T2) and two conserved domains (Pru-C2 and Pru-C4R) of sweet cherry S-RNases. By comparing the size of PCR products on agarose gel, the 5-genotypes of ‘Kouka-Nishiki’, ‘Beni-Sayaka’, ‘Beni-Shuho’ and ‘Merton Glory’ were suggested to be S¹S³, S¹S⁶, S⁴S⁶, and S⁴S⁶, respectively. Two of these three S-genotypes (S¹S⁶ and S⁴S⁶) were found for the first time. DNA sequencing of PCR products from S-alleles of ‘Rainier’ and ‘Sato-Nishiki’ revealed that Ban II, Nru I, Apa LI and Ava I sites, respectively, were unique in the S¹-, S³-, S⁴- and S⁶- sequences flanked by Pru-T2 and Pru-C4R primers. RFLP analysis of the PCR products using these enzymes confirmed that S¹-, S³-, S⁴- and S⁶-alleles of the four cultivars contained the respective restriction enzyme recognition sites.     

S_allele identification and genetic diversity analysis of apricot cultivars

In this study, the sexual incompatibility and S-allele diversity of 24 Turkish apricot cultivars, Paviot and Sak?t-1 as parents and 127 F₁ progenies were identified by polymerase chain reaction (PCR) and sequencing techniques. Additionally, genetic diversity and relatedness among the 24 cultivars were determined using 18 simple sequence repeat (SSR) markers from the genus Prunus. PCR for S_alleles identified nine different S-RNase alleles in the 24 apricot cultivars, namely S_c, S₁, S₂, S₈, S₉, S₂₀, S₂₄, S₅₂, and S₅₃. All primers amplified only one S_allele in the cultivars Adilcevaz-1, Adilcevaz-3, Ethembey, Pasamismisi, Canakkale, and Soganci. Most of the Turkish cultivars were self-incompatible. The S_c allele was present in only three cultivars (Canakkale, Ethembey, Imrahor) that are, therefore, self-compatible. The S_alleles of cultivars Paviot and Sak?t-1 displayed homology with the S_c, S₂ and S₂₀, and S₅₂ alleles. In the 127 F₁ genotypes, the two S_alleles of Paviot were inherited by roughly half of the offspring, while about 76% of the offspring inherited the S₅₂ allele from Sak?t-1, and less than 24% inherited S₂₀. The amplification using all SSR 18 primers was successful and produced 128 polymorphic alleles with an average of 7.11 alleles per locus. Among the apricot cultivars studied, expected heterozygosity (He) ranged from 0.33 to 0.72, observed heterozygosity (Ho) ranged from 0.42 to 1.00, PIC values were between 0.28 and 0.89, and similarity rates were between 0.30 and 0.68. The cultivars Levent and Ozal were genetically closest (0.68) while cultivars Sak?t-3 and Soganc? were the most distinct (0.30).     

Analyses of pollen-tube growth and biological action of S-RNase in the style of self-compatible Japanese pear

The Japanese pear ‘Osa-Nijisseiki’ (S₂S₄^SM) (SM = stylar-part mutant) is a self-compatible bud mutant that originated from self-incompatible ‘Nijisseiki’ (S₂S₄). The S₄-allele of the pear is deleted and it shows unilateral incompatibility to cultivar with an S₂S₄ genotype. However, when pollen-tube growth was compared between cross-compatible [‘Osa-Nijisseiki’ × ‘Okusankichi’ (S₅S₇)], unilateral-compatible [‘Osa-Nijisseiki’ × ‘Kikusui’ (S₂S₄)], self-compatible (‘Osa-Nijisseiki’ × ‘Osa-Nijisseiki’), and unilateral-incompatible pollination (‘Kikusui’ × ‘Osa-Nijisseiki’), pollen-tube growth clearly showed the following order: cross-compatible > unilateral-compatible > self-compatible > unilateral-incompatible. This indicates that the ‘Osa-Nijisseiki’ style produces specific inhibitor(s) not only to S₂- and S₄-pollen but also “self-pollen”, because the phenotype of S₄^SM-pollen is the same as S₄-pollen. Stylar protein analysis demonstrated that ‘Osa-Nijisseiki’ produces S₂-RNase (RNase associated with S₂-allele) together with a small amount of S₄-RNase. The purified S₄-RNase possessed almost the same inhibitory action on the growth of S₄-pollen-tubes in vitro at 1 μg μl⁻¹ as that from original ‘Nijisseiki’. These results suggest that the depressed growth of unilateral-compatible and self-pollen-tubes in ‘Osa-Nijisseiki’ is due to this biologically active S₄-RNase. Growth of self-pollen-tubes may also be depressed by inhibitor(s) specific to “self-pollen” unrelated to S-alleles.     

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-genotyping of 25 sweet cherry (Prunus avium L.) cultivars from the Czech Republic

Sweet cherry (Prunus avium L.) is a self-incompatible species. Determination of the S-genotypes of cherry cultivars is crucial for breeding and to select appropriate cultivars for cross-fertilisation and fruit set. In this study, we characterised the S-genotypes of 25 sweet cherry cultivars, some of which had being bred at the Research and Breeding Institute of Pomology (RBIP), Holovousy, Czech Republic, and others were European cultivars in the RBIP collection. S-genotyping was carried out by polymerase chain reaction using consensus primers for the S-RNase and SFB genes, and capillary electrophoresis. Nine different known S-haplotypes (S₁, S₂, S₃, S₄, S₅, S₆, S₉, S₁₃, and S₁₆) were identified and the cultivars were assigned to 12 incompatibility groups. One local cultivar, ‘Pta?ka z Plzně’, originated from a wild forest seedling and used as a pollinator, was assigned to Group 0 of universal donors. The pedigree of some cultivars was confirmed by their S-genotype. This study represents the ?rst comprehensive S-genotype screening of sweet cherry genetic resources in the Czech Republic and will be useful for the design of crossing programmes and orchard management of these sweet cherry cultivars.     

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.     

Characterisation of the chemical composition of scab-resistant apple pomaces

Summary

Pomaces obtained during the production of cloudy or clear juice from scab-resistant apple cultivars (28 harvested in 2006, and 23 harvested in 2007) were studied as a source of nutritionally important components. It was shown that the average yield of pomace during the production of cloudy juice was 6.4% (w/w), and 4.9% (w/w) for clear juice pressing. The average total dietary fibre (TDF) contents were 48.4% (w/w) and 52.4% (w/w) for cloudy and clear juice pomaces, respectively. Ten pomaces from cloudy apple juice production, and 12 obtained from clear juice production, contained ≥ 2,000 mg kg^?1 of polyphenols.‘Ariwa’ and ‘Rajka’ are apple cultivars suitable for juice production and their pomaces are characterised by having high contents of TDF [50.9% (w/w) and 51.1% (w/w), respectively], with aboveaverage polyphenol and quercetin glycoside concentrations. The mean contents of quercetin glycosides for pomaces from all apple cultivars was 994 mg kg^?1 after clear juice production, and 908.2 mg kg^?1 after cloudy juice production. ‘Topaz’, which became a popular commercial apple cultivar in Europe, was the best source of health-promoting components. Its pomace was the richest source of polyphenols among all 28 cultivars studied. It contained ≥ 2,000 mg kg^?1 quercetin glycosides in both its clear and cloudy juice pomaces.     

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.     

Chemical composition of French and Polish cloudy apple juices

Summary

Cloudy juices from six apple cultivars from Poland (‘Ariwa’, ‘Gold Milenium’, ‘Florina’, ‘Melfree’, ‘Novamac’, and ‘Rajka’) and four French cultivars (‘Ariane’, ‘Chanteline’, ‘Judeline’, and ‘Judor’) were produced and chemically characterised. The analyses encompassed 23 chemical parameters and phenolics profiles. The most important parameter, differentiating cloudy juice from clear juice, was turbidity. Cloudy juices were characterised by having an average total turbidity of 1,210 Nephelometric Turbidity Units (NTU) and a stability of turbidity of 42%. Some of the results deviated from the accepted ranges given in reference values for apple juices in the Code of Practice of the European Fruit Juice Association. This occurred in the cases of simple sugars and saccharose contents of the Polish apple juices (e.g., up to 56.9 mg l^–1 of saccharose vs. a maximum value of 30 mg l^?1), and for some mineral compounds in the French apple juices (e.g., sodium values up to 10 mg l^?1). Large variations were found in case of important healthconferring components. Water-soluble pectin contents varied from 205 mg l^?1 for ‘Chanteline’, to 1,289 mg l^?1 for ‘Gold Milenium’; while, in the case of phenolic compounds, the range was from 85.7 mg l^?1 for ‘Novamac’, to 524.8 mg l^?1 for ‘Melfree’.     

The Pillnitz Re-Series of Apple Cultivars – Do They Hold Promise? – 80 Years of Professional German Fruit Breeding 2008

The aim of the Pillnitz apple breeding is to combine improvements in fruit quality + yield + resistance to different pathogens in new cultivars. Early-, mid- and late-season apple cultivars were selected in two series, Pi- and Re-cultivars?. The Re-cultivars? guarantee a high degree of resistance. Triple and multiple resistant cultivars were selected with resistance to scab, mildew and fire blight: ‘Remo^®’, ‘Regia^®’, ‘Rewena^®’ and ‘Rebella^®’. ‘Rebella^®’ was found to have resistance also to bacterial canker, red spider mite and abiotic damages. The cultivars are the base for a new growing management in integrated and organic cultivation. Doubtless, the Pillnitz resistance breeding conception is a success, even if some problems remain to solved in the future. Various degrees of scab infection on Vf-resistant cultivars had been observed since 1984 in Central Europe. No infections were found in cultivars with other genes like V _f such as ‘Reglindis^®’ (V _A), ‘Reka’ (V _r), or ‘Regia^®’ (V _r). New sources are necessary for resistance breeding, especially for pyramiding of resistance genes. However, if the V _f-gene is overcome, new resistance sources and cultivars with two or more different sources of resistance to stabilize healthiness in the field will be required in the future. More explanation and advertisement about the value of resistant cultivars to promote the cultivation in commercial orchards is needed.     

京白梨等品种S 基因型鉴定及新基因S28 和S30 的核苷酸序列分析

 根据梨S基因高度保守区C1和C3区, 设计1对引物P1和P2, 对梨品种的基因组DNA进行S基因特异扩增、克隆、测序, 并在GenBank中BLAST比较, 确定S 基因特异性片段, 对京白梨等6个供试自交不亲和品种的S基因型比对结果为: 白梨中的‘库尔勒香梨’为S₂₁ S₂₈ , ‘苹果梨’为S₁₇S₁₉ ; 砂梨中的‘台湾蜜梨’为S₁₁ S₂₂ ; 西洋梨中的‘葫芦梨’为S_a S_b; 秋子梨中的‘京白’为S₁₆ S₃₀ , ‘早梨18’为S₄ S₂₈。其中S₂₈和S₃₀为首次登录的新S 基因, 在GenBank的登录号分别为AY562394 (库尔勒香梨) 和AY876945 (京白) 。     

The Effect of ‘Latent’ Virus Infections on the Yield of Maiden Trees on 20 Apomictic Apple Seedling Rootstocks

Five years’ experiments with apomictic apple seedling rootstock selections budded with apple cultivars have shown that incompatibility in Malus sieboldii, M. sargenti, M. hupehensis and their derivatives is mainly due to high sensitivity of the species rootstocks to viruses latent in commercial apple cultivars. The yields of maiden trees from rootstocks which were budded with material that was virus-free, infected with ‘latent’ viruses and infected with ‘latent’ viruses plus rubbery wood virus were 67%, 28% and 16%, respectively. There were, however, differences in the sensitivity reaction between the rootstocks tested. Fairly good compatibility with scions infected with ‘latent’ viruses was observed in the M. sieboldii root- stock selections 4542, 4556 and 4608 and in M. hupehensis.     

Self- and cross-(in)compatibility between important apricot cultivars in northwest Iran

Summary

Knowledge of the self-(in)compatibility trait in commercial apricot cultivars is of great importance for breeders and growers. Five commercial apricot cultivars, widely grown in Iran, were self- and cross-pollinated to determine their pollen and stylar compatibility. Fruit-set in the orchard and pollen tube growth in pistils, from flowers pollinated in the laboratory, were evaluated. In addition, specific primers previously designed to amplify fragments of the S alleles responsible for the incompatibility trait, were used to amplify DNA extracted from the five cultivars.All results agreed and confirmed that three out of the five cultivars studied were self-incompatible, two of which were cross-incompatible and therefore had the same genotype. The cultivars, ‘Ghorban-e-Marageh’ and ‘Ghermez-e-Shahroodi’ were self-compatible and, interestingly, shared a PCR band with all Spanish self-compatible apricot cultivars examined to date.     

Effects of NAA and Carbaryl on Fruit Set and Return Bloom for Some Apple (Malus domestica Borkh.) Cultivars

Abstract

This research was conducted in 1997 and 1998 to investigate the effects of naphthalene acetic acid (NAA) (5, 10, and 15 ppm), Carbaryl (1-naphthyl N-methylcarbamate) (750, 1000, and 1250 ppm), and NAA + Carbaryl (5 + 750, 7.5 + 750, and 10 + 750 ppm) applications on the return bloom of some standard apple (Malus domestica Borkh.) cultivars grafted on MM106 rootstock. Of these applications, 750 ppm Carbaryl for ‘Starkspur Golden Delicious’, 1250 ppm Carbaryl for ‘Granny Smith’ and ‘Starkrimson Delicious’, and 5 ppm NAA for ‘Jonagold’ increased the mean number of flower buds significantly, compared with the control treatments. The other treatments of Carbaryl, NAA, and NAA + Carbaryl also increased the mean number of flower buds in a nonsignificant sense with a few exception. A negative correlation between the final fruit set and the mean number of next year's flower buds was found for three cultivars. The correlation coefficients were r = ?0.5150 (P< 0.05), r = ? 0.6999 (P< 0.05), r = ?0.0335 for ‘Starkspur Golden Delicious’, ‘Granny Smith’, and ‘Jonagold’ cultivars, respectively. However, this relationship was positive and nonsignificant for ‘Starkrimson Delicious’ (r = 0.1980).