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A critical evaluation of methods used for S-genotyping: from trees to DNA level
19-29.Views:208Fruit setting behaviour of fruit trees remains to be in the focus of plant breeders and growers. Realizing that most species (cherry, apple, pear etc.) are self-incompatible and certain cultivars are cross-incompatible, mutual fertility properties and their reliable determination are of great interest. This review gives a comprehensive description of all known S-genotyping procedures, i.e. the classical fruit set analysis after open field test crosses; pollen tube growth monitoring with fluorescent microscopy; stylar ribonuclease electrophoresis (using different types of isoelectric focusing and 2-dimension polyacrilamide gel electrophoresis); as well as the most recent polymerase chain reaction based DNA-level analyses and DNA sequencing. The review presented not only gives a compilation of the bases of the methods described but also provides a critical evaluation and a comparative characterization of their applicability.
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Preliminary evaluation of breeding perspectives of Ukrainian sweet cherry cultivars: nutraceutical properties and self-incompatibility
7-11.Views:352Some traditional sweet cherry cultivars of Ukrainian origin may represent perspective material for Hungarian cherry breeding. A total of eight cultivars analysed represent great diversity in several phenotypic traits including fruit ripening time or fruit flesh colour. Considerable differences in the anthocyanin content may result in different antioxidant capacity of fruits. In the present study, we used ferric reducing antioxidant power (FRAP) and total phenolic content (TPC) assays to characterize fruits’ nutraceutical properties. These values were compared with the respective values measured for eight commercial cultivars grown in Hungary. The average of FRAP and TPC values was higher for the Ukrainian cherries compared with commercial cultivars suggesting they might be included in functional breeding programs. Since, cherry is a self-incompatible species, the determination of S-genotype is required for both breeding and successful cultivar association in commercial orchards. Complete or partial S-genotypes were determined for 5 and 3 cultivars, respectively.
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Review of the molecular background of self-incompatibility in rosaceous fruit trees
7-18.Views:235This review gives a presentation of the gametophytic self-incompatibility system in the roscaeous fruit trees. Studies to discover the pistil (S-ribonucleases) and pollen-part components (F-box molecules) are summarized and models for the self-incompatibility reactions as well as their molecular background are discussed. We describe how mutations within the S-RNase or F-box genes can contribute to the transition from self-incompatibility to the self-compatible phenotype in many fruit tree crops. The current state of the arts is compared to the information obtained in other plant species possessing similar incompatibility system.
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Characterization of quince (Cydonia oblonga Mill.) cultivars using SSR markers developed for apple
7-10.Views:370Quince (Cydonia oblongaMill.) is a minor fruit crop, which is primarily used for marmalade, jam and sauce.Very few quince cultivars are known all over the world and in many cases similar names are used for presumably different cultivars. The aim of the present study was to evaluate and characterize the genetic diversity of 36 quince cultivars and selections with SSR markers. Seven out of 8 SSR markers designed from apple sequences could successfully yield amplification also in quince cultivars. Number of alleles per locus ranged from 2 to 3 alleles. These allele numbers are quite low when compared to apple. It is supposed to be the consequence of a genetic bottleneck. In spite of the low allele number per locus, the 36 quince cultivars formed 30 different genotypes. The ratio of homozygosity was low, which might be coupled with the self-(in)compatibility phenotype of quinces. SSR markers proved unable to differentiate putatively closely related cultivars (e.g. ‘Bereczki’ and ‘Bereczki bôtermő’). In general, the level of polymorphism among the tested quince genotypes was much restricted due to the low allele number detected. However, it must be considered that the number of analysed SSR loci is not enough high to estimate the overall heterozygosity of the quince genome. Further experiments are needed and the SSR markers proved to be a reliable and useful tool for such analyses.
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Genotyping Hungarian apricot cultivars for self-(in)compatibility by isoelectric focusing and PCR analysis
69-72.Views:166Self-incompatibility (SI) in flowering plants is a widespread genetic system that promotes out-crossing. In Prunus species the SI is a gametophytic trait, which is controlled by a single multiallelic locus, termed S-locus. S-alleles codify stylar glycoproteins with ribonuclease activity (S-RNases). Our objective was to assess the S-genotype of some Hungarian apricot varieties by isoelectric focusing of stylar RNases as well as by PCR analysis using cherry consensus primers. Consensus primers amplified one or two bands of various sizes. Primers amplifying the 1st intron gained fragments the size of which ranged from 250 to 500 bp; while those amplifying the 2nd intron resulted in fragments of 800-2000 by length. Our data demonstrated that the first intron of the apricot S-RNase gene is shorter than the second one, which coincides with the structure of cherry S-RNase alleles. `Hargrand' (S1S2) and `Harcoe (S1S4) possessed one common S-RNase isoenzyme. Hungarian 'Orias' apricot cultivars showed different bands compared to the previous cultivars, but they shared completely identical patterns confirming that they possess the same S-genotype. 'Bergeron', `Harmat' and 'Korai zamatos' are characterised by an evidently distinct S-RNase pattern. The self-compatible cultivar (`Bergeron') had one allele, which suggests its correspondence to the Sc. Primers for the 2nd intron was unsuccessful in gaining fragments, which indicates that the 2nd intron in the Sc allele is too long to get any amplification. On the basis of our data, identities and differences were revealed in the S-allele constitution of some economically important Hungarian apricot cultivars at protein and DNA levels.
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Optimization of RNA isolation from stone fruits at different ripening stages
101-104.Views:205This study was conducted to select the most appropriate RNA isolation method that can be used successfully in case of stone fruits. The changing pattern of gene expression during the ripening process of stone fruits may elucidate the molecular background of several phenotypical or phytochemical alterations present among different genotypes. Our laboratory aims to study the expression of genes encoding for enzymes that catalyze crucial steps in the flavonoid biosynthesis pathway. RNA isolation from fruit mesocarp is a challanging task due to high levels of sugars and polyphenolics accumulating during fruit development. Therefore, at first, the optimal techniques eligible for RNA isolation from fruit tissues at different ripening stages must be selected. Our study compares three different RNA isolation protocols and describes their potential applicability according to different fruit species and ripening stages.
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Pollen tube growth in sweet cherry (Prunus avium L.) styles following fully compatible, half compatible and incompatible pollinations
63-68.Views:142In vivo as well as in vitro pollen tube growth studies along the style were performed, each with two pairs of sweet cherry cultivar combinations by means of fluorescence microscopy. In vivo studies showed that the percentage of pollen tubes penetrating the middle and basal section of the style was higher in the fully compatible 'Margit' x 'Alex' combination than in the half compatible `Germersdorfi 3' x `Alex' cross. The year effect was significant at P=0.] probability level. All pollen tubes in vitro stopped at the upper third of the style in the incompatible 'Vera' x 'Van' cross, whereas in the half compatible 'Alex' x 'Van' 50% of the pollen tubes penetrated to the lower third of the style. By in vitro fluorescence microscopy, it was possible to distinguish half compatible combinations from incompatible ones. Results obtained by in vivo technique only were much ambiguous.
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Recent findings of the tree fruit self-incompatibility studies
7-15.Views:217This review endeavours to collect all recent and substantial contributions to the quickly deepening fields of tree fruit self-incompatibility studies and hence updating previously published reviews. Studies carried out to discover the molecular basis of gametophytic self-incompatibility are summarized and a newly described model for the solanaceous plants is also outlined. We describe recent findings in all economically important fruit tree crops involving apple, European pear, sweet and sour cherries, almond, Japanese plum, sloe, Japanese apricot, European apricot and peach. Additional DNA sequences are now available for both the pistil and pollen component genes in several species and their molecular, evolutionary or economic implications are discussed in the light of the fruit setting behaviour.
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Preliminary characterization of the self-incompatibility genotypes of European plum (Prunus domestica L.) cultivars
23-26.Views:234European plum is an important fruit crop with complex, hexaploid genome of unknown origin. The characterization of the selfincompatibility (S) locus of 16 European plum cultivars was carried out using the PaConsI-F primer in combination with the EM-PC1consRD primer for the first intron and the EM-PC2consFD and EM-PC3consRD primers for the second intron amplification. Altogether, 18 different alleles were scored indicating high genetic diversity. These alleles were labelled using alphabetical codes from SA to SS. We identified 5 different alleles in 9 cultivars, 4 alleles in 5 cultivars, while 3 alleles were shown in two of the assayed cultivars. A total of 16 different S-genotypes were assigned, and discrimination of all plum cultivars was successful based on their unique S-genotypes. However, further research is required to reliably identify the S-alleles based on their DNA sequence and clarify complete S-genotypes.
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RAPD analysis of grapevine hybrids and cultivars
63-66.Views:168Utilization of the Randomly Amplified Polymorphic DNA (RAPD) technique as a molecular marker was tested to investigate the relationships between some representative grapevine cultivars and hybrids established at the Department of Genetics and Plant Breeding (CUB), to distinguish clones as well as to characterize various hybrids between species or cultivars and their parents. Vitis vinifera cultivars were easily and successfully distinguished by the RAPD technique and they were grouped according to the traditional taxonomic classification. RAPD patterns of the examined Pinot gris clones proved to be completely identical. Number of generations was reflected by the value of genetic distance of the examined hybrids. Genetic identity of parents and their offsprings was influenced by the selection applied in the process of plant breeding. Parental phenotypic and morphologic characteristics showed high degree of segregation in hybrids, but RAPD analysis revealed that their genetic similarity is considerable. The three Vitis anntrensis clones were properly discriminated from every cultivar and hybrid of Vitis vinifera, i.e. hybrids are much closer to the cultivated grapevine than to V. anzurensis due to the phenotypic selection carried out during the life-cycle of one or two generations.
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Self-incompatibility in plums (Prunus salicina Lindl., Prunus cerasifera Ehrh. and Prunus domestica L.). A minireview
137-140.Views:490Japanese plums (P salicina) and cherry plums (P cerasifera) are diploid species, while European plum (P. domestica) cultivars are hexaploids. Most diploid species are self-incompatible while fertility relations of the hexaploid European plums are variable between self-incompatibility and self-compatibility. About twenty S-alleles and six inter-incompatibility groups and one S-haplotype responsible for the self-fruitful phenotype were described in Japanese plum cultivars, but studies on cherry plums and even on the European plum cultivars are severely restricted. This review is focused on the available information obtained from myrobalans and European plums; and discusses recent hypotheses regarding the putative origin of the hexaploid plums, and thereby indicates the possibility of allele flow between different plum species.
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S-locus genotyping on stone fruits in Hungary: a review of the most recent achievements
39-43.Views:210Central Europe can be taken as a geographical and historical connection zone between the western growing countries and Asian gene centres of Prunus tree fruits. The determination of the S-genotype of stone fruit (mainly almond, plum, cherries and apricot) cultivars and landraces has both practical and theoretical significance. Our group has allocated complete S-genotypes for more than 200 cultivars and selections of almond, Japanese plum, sweet cherry and apricot. Among Eastern European almond cultivars, two novel cross-incompatibility groups (CIGs) were identified. S-alleles of a related species were also shown in P. dulcis accessions; a fact seems to be indicative of introgressive hybridization. Our results with Japanese plum clarified and harmonized two different allele nomenclatures and formed a basis for intensive international studies. In apricot, a total of 13 new S-alleles were identified from Eastern European and Asian accessions. Many Turkish and North African cultivars were classified into new CIGs, III–XVII. Results suggest that the mutation rendering apricot self-compatible might have occurred somewhere in south-east of Turkey and we were successful to confirm the presumed Irano-Caucasian origin of North African apricots based on the geographical distribution of S-alleles. In sweet cherry, new alleles have been identified and characterized from Turkish cultivars and selections. In addition, wild sweet cherry and sour cherry S-alleles were also shown indicating a a broader gene pool in Turkey as compared with international cultivars. We also used S-genotype information of Ukrainian sweet cherry cultivars to design crosses in a functional breeding program. Our results exhibit an increased number of S-alleles in tree fruit accessions native to the regions from Eastern Europe to Central Asia, which can be used to develop S-genotyping methods, to assist cultivation and draw inferences for crop evolution.
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Self-incompatibility in pears (Pyrus communis L., Pyrus serotina Rehd. and Pyrus ussuriensis) Review
87-91.Views:206Self-incompatibility system and allele pool of three different pear species, European pear (Pyrus communis), Japanese pear (P. serotina) and Chinese pear (P ussuriensis) are displayed. Several inconsistencies and the absence of the harmonization of three different allele series are revealed in the European pears. By collecting data from several reports eight incompatibility groups of Japanese pear cultivars could be established. A self-compatible genotype is analysed in details and shown to be a stylar-part mutant. As Japanese pear was the first fruit tree species from which S-ribonucleases were identified, the history of S-genotyping from the beginning to the latest achievements and technical developments can be also monitored from the experiments enumerated. In Chinese pears, seven S-alleles and one incompatibility group could be identified.
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Self-incompatibility alleles in Esatern European and Asian almond (Prunus dulcis) genotypes: a preliminary study
23-26.Views:202Almond [Prunus dulcis (Mill.) D. A. Webb.] as one of the oldest domesticated plants is thought to have originated in central Asia. Gametophytic self-incompatibility of almond is controlled by the highly polymorphic S-locus. The S-locus encodes for an S-ribonuclease (S-RNase) protein in the pistils, which degrades RNA in self-pollen tubes and hence stops their growing. This study was carried out to detect S-RNase allelic variants in Hungarian and Eastern European almond cultivars and Turkish wild growing seedlings, and characterize their S-allele pool. Five new alleles were identifi ed, S31H, S36-S39 in Eastern European local cultivars. The village Bademli and Akdamar island are two distinct places of almond natural occurrence in Turkey. Trees growing wild around Bademli city showed greater genetic diversity than those originated on Akdamar island. Many of the previously described 45 S-RNase alleles have been also detected in these regions. Homology searches revealed that Turkish almonds carried some P. webbii alleles indicating hybridization between the two cultivars and massive introgression events. Our results supply long-awaited information on almond S-allele diversity from regions between the main cultivation centres and the centre of origin of this species; and are discussed from the aspect of methodological developments and evolution of the cultivated almond.