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1.
Sweet cherry (Prunus avium L.) has stylar gametophytic self‐incompatibility, which is controlled by the multi‐allelic S‐locus and encompasses the highly polymorphic genes for the S‐ribonuclease (S‐RNase) and S‐haplotype‐specific F‐box (SFB), which are female and male determinants, respectively. The self‐compatible mutant SFB4′ corresponds to an allele variant of SFB4 and presents a frameshift mutation. Even though male‐determinant molecular markers can discriminate between SFB4 and SFB4′ alleles, the methods required are laborious, time‐consuming and expensive, and not suitable for massive analysis and integration into breeding programmes. Our aim was to develop molecular markers for the evaluation of self‐compatibility alleles in sweet cherry, that could be used as a high‐throughput screening strategy to identify SFB4 and SFB4′ alleles, based on a marker for male determinacy. Our results were consistent using primers flanking the mutation responsible for the SFB4′ allele. We designed a specific molecular marker and confirmed it in sweet cherry commercial varieties. This new molecular marker is feasible for self‐compatibility alleles in the male determinant in sweet cherry‐assisted breeding programs.  相似文献   

2.
Primers amplifying a range of Prunus S-alleles   总被引:2,自引:1,他引:2  
Although various consensus polymerase chain reaction (PCR) primers have been reported for identifying Prunus S‐alleles, they have been developed from and optimized on a limited set of alleles, which may limit their applicability to a broader allele range. To develop a primer set for use across the genus, degenerate consensus primers were designed from conserved regions of 27 S‐RNase sequences available from five Prunus species. The primers were tested in 15 previously genotyped cultivars of cherry, almond and apricot, representing alleles S1 to S6 in each crop and also Sc in apricot. Comparisons were made with previously published primers tested in the same 15 cultivars under reported reaction conditions. The new primers generated an amplification product for each of the 19 S‐alleles whereas those previously available amplified no more than 14. The primers will be useful for genotyping and genetic studies in cultivars and wild populations.  相似文献   

3.
M. López    M. Romero    F. J. Vargas    M. Mnejja    P. Arús    I. Batlle 《Plant Breeding》2005,124(5):502-506
To verify the compatibility behaviour of the almond cultivar ‘Francolí’ and to clarify its S genotype a combination of pollination tests, stylar ribonuclease and allele specific PCR analysis was used. ‘Francolí’ was released from IRTA's breeding programme in 1994, having been putatively raised from the cross ‘Cristomorto’ (S1S2) × ‘Gabaix’ (S10S25). This cultivar was also reported to be self‐incompatible but revealing only one S band in the zymograms after S‐RNases analysis. ‘Francolí’ sets nuts after test crossing with two S1S25 cultivars, having a different genotype from that earlier reported. ‘Francolí’ was also observed to be self‐compatible after selfing flowers in the field and in the laboratory. ‘Francolí’ was re‐assigned the S1Sf genotype after test crossing, stylar ribonuclease and PCR data analysis. After microsatellite analysis, the self‐compatible ‘Tuono’ (S1Sf) cultivar is suggested as the male parent of ‘Francolí’ instead of the earlier reported ‘Gabaix’.  相似文献   

4.
S-allele identification by PCR analysis in sweet cherry cultivars   总被引:3,自引:0,他引:3  
Gametophytic self‐incompatibility, governed by the S‐locus, operates in sweet cherry. The knowledge of the S‐genotype of sweet cherry cultivars is therefore essential to establish productive orchards by defining compatible combinations. The isolation of sweet cherry S‐R Nases has allowed the use of different molecular techniques to characterize the S‐genotypes of sweet cherry cultivars. Previously, incompatibility group assignment could only be carried out on mature trees through pollination tests. In this work, PCR analysis with primers designed on the conserved sequences of sweet cherry S‐R Nases has been used to characterize the S‐genotype of 71 sweet cherry cultivars, including 26 cultivars whose S‐allele constitution had not been previously described. This approach has allowed the detection of alleles that had not been amplified by PCR before, to identify six putative new S‐alleles, to define three new self‐incompatibility groups and to compile the standards for a PCR‐based S‐allele typing method in sweet cherry.  相似文献   

5.
Almond is a highly heterozygous species with a high number of S‐alleles controlling its gametophytic self‐incompatibility system (GSI). In this work, we have analysed 14 Spanish local almond cultivars for S‐RNase allele diversity. Five new S‐RNase alleles were identified by cloning and sequencing, S31 (804 bp) in ‘Pou de Felanitx’ and ‘Totsol’, S32 (855 bp) in ‘Taiatona’, S33 (1165 bp) in ‘Pou d’Establiments’ and ‘Muel’, S34 (1663 bp) in ‘Pané‐Barquets’ and S35 (1658 bp) in ‘Planeta de les Garrigues’. Additionally, seven already known almond alleles could be recognized in the local cultivars studied. The high number of new alleles identified reveals the wide diversity of almond germplasm still existing and requiring characterization, and points to the possibility of new findings by a wider study focusing on other provenances. The almond S‐RNases have been compared to those of other Prunus species, showing a high identity and confirming that the S‐RNase gene in this genus presents a probable common ancestor.  相似文献   

6.
Most cultivars that belong to the Rosaceae are self‐incompatible and depend on cross‐pollination. The pollen donor and pollen recipient have to flower synchronously and must be genetically compatible. Genetic compatibility is governed by the S‐locus, which holds the S‐RNase and S‐haplotype‐specific F‐Box (SFB) genes. Thus, the S‐genotype of cultivars is an important feature and is characterized molecularly by the S‐RNase and SFB alleles which are distinctive for each S‐haplotype. Here, we report the usage of DNA chromatography (denaturing high‐performance liquid chromatography – DHPLC) for identifying the S‐genotypes of European apricots on the basis of their SFB alleles. DHPLC is amenable to high‐throughput automation, and therefore is valuable for breeding and for high‐quality plant typing in the nursery.  相似文献   

7.
Prunus mandshurica [(Maxim.) Koehne] from Central Asia is a species related to apricot and grown in China and Mongolia. This species has been used in apricot breeding as a source of frost resistance. In addition, P. mandshurica has been suggested as the possible origin of some North American apricot cultivars resistant to sharka (Plum pox virus, PPV). The aim of this work was to transmit the resistance to PPV from P. mandshurica to the Spanish apricot cultivar ‘Currot’ by traditional crossing. The resistance to a Dideron PPV isolate of the descendants of P. mandshuricaבCurrot’ and their progenitors was evaluated under controlled conditions in a greenhouse. The results showed the susceptibility of both, the progenitors and the offspring to PPV, as being much higher than in other apricot seedlings. The results showed that the P. mandshurica accession studied is not a good progenitor in breeding apricot for PPV resistance, but just the opposite. The possible role of P. mandshurica as a source of resistance in apricot resistant cultivars is questioned.  相似文献   

8.
The stylar products of the S‐locus for the gametophytic self‐incompatibility (GSI) system in the Rosaceae are ribonucleases (S‐RNases). Recently, sequences for 13 pear S‐RNase alleles have been published and named following a letter–symbol nomenclature (Sa to Sd and Sh to Sp). To establish the correspondence between these sequences and the self‐incompatibility alleles we have described previously (S1 to S5), we have amplified genomic DNA with consensus primers from the cultivars, ‘Williams’ (S1S2), ‘Coscia’ (S3S4), ‘Butirra Precoce Morettini’ (S1S3), ‘Santa Maria Morettini’ (S2S3) and ‘Doyenne du Comice’ (S4S5) and identified PCR products specifically associated with each S allele. Cloning and sequencing of the amplification products has revealed that they correspond to European pear sequences already deposited in the database. This allowed us to link S‐RNase sequences with S allele phenotypes and to determine a correspondence between the symbol–letter nomenclature used to name S‐RNase sequences and the number‐based nomenclature used to name S alleles. Based on this result the prediction of new cross‐incompatibilities among pear cultivars is discussed. Finally, we propose a unified number‐based nomenclature to avoid future confusion denominating S alleles in pear.  相似文献   

9.
10.
self-(in)compatibility almond genotypes: A review   总被引:1,自引:0,他引:1  
To compile self-(in)compatibility almond genotypes, a review of 133 commercial cultivars of wide geographical origin was made. The information gathered from own and mainly published work will be useful for both grower's cultivar choice when planting and for breeder's cross design when planning. The almond S genotypes compiled were identified using five different methods: biological (pollination tests in the field and in the laboratory) and molecular (RNases, PCR and sequencing). In most cases, genotypes were assigned after combining more than one technique. Cultivars were classified into three categories: self-incompatible (99), self-compatible (16) and doubtful self-incompatible (18). The database is divided in 9 fields (name, origin, parentage, obtention year (crossing, selection or release), S genotype, technique used, reference, consensus genotype, and cross incompatibility group). A study of the 27 S alleles already identified and their geographical distribution within the cultivated almond is also presented. The study was divided into cultivars of known and unknown parentage and the distribution of S alleles frequencies was uneven among the 133 cultivars. S allele frequencies are related to geographical origin. Some alleles (S 1, S 5, S 7 and S 8) are more frequently observed than the others among cultivars of both known and unknown parentage. In the cultivated almond, the S f allele is only found in the Puglia region, Italy. The S f frequency is three times higher in cultivars released from breeding programmes than in cultivars selected by growers. From the 351 resulting possible genotypes by combination of the 27 S alleles identified only 20 CIG (0-XIX) have been established, which represents a small fraction of the whole genetic diversity of this polymorphic gene in almond.  相似文献   

11.
Italian almond germplasm is characterized by a wide diversity in several growing areas among which Sicily is one of the most important. Analysis with consensus and specific primers and DNA sequencing was performed to investigate S‐RNase genetic diversity and to elucidate the homology rate within a genetic pool of 27 Italian accessions. Interestingly, some of the self‐compatible cultivars did not show the presence of Sf allele. Amplicons from consensus and allele‐specific PCR primers revealed a high level of variability. Sequencing of all the S‐RNase amplicons derived from consensus primers allowed the identification of two new S‐RNase alleles (S51 and S52). Surprisingly, despite the AA replacement mutation, S51 did not exhibit any change of its S‐RNase function. Additionally, several mutations, with no effect on amino acid composition, were detected in the intron and/or in the ORF of four known alleles (Sg, S10, S31 and S35). Genetic variation, regarding point mutations and only detected by sequencing, was revealed among 11 of 27 tested cultivars. The new sources of variability might have an interest for product traceability.  相似文献   

12.
To clarify incompatibility relationships among almond cultivars, 35 were analysed for stylar ribonucleases, which have previously been shown to correlate with incompatibility S alleles. Stylar proteins were extracted and separated electrophoretically and the zymograms compared with ladders of ribonucleases corresponding to the 12 S alleles previously reported. Sixteen cultivars showed a band corresponding to two of the known ribonucleases, 17 showed one known ribonuclease and one ‘new’ band, and two showed two new bands. Twelve new ribonucleases were detected; 11 were attributed to new S alleles (S13 to S23) and a mutant form of S7 was attributed to S7A. Genotypes were proposed for nine cultivars of five incompatibility groups that had not been genotyped previously, VII, X, XI, XII and XIII. Twenty‐four cultivars of unknown incompatibility relationships were provisionally genotyped: six of these could be assigned to existing groups and two new groups were established, XIV and XV, along with group O of cultivars with unique genotypes. Test crosses confirmed that eight pairs of cultivars showing similar zymograms were indeed cross‐incompatible, including the two representatives of each of the two new groups. Virtually all self‐incompatible cultivars of known genotype are listed in a table. The data should be useful for planning cultivar combinations for orchards and for designing crosses for breeding programmes.  相似文献   

13.
The work aimed to develop a reliable and convenient PCR approach for determining incompatibility S genotypes in almond. Initially, genomic DNAs of 24 accessions of known S genotype were amplified with novel consensus primers flanking the first and second introns of the S‐RNase gene. The PCR products separated on agarose showed length polymorphisms and correlated well with the reference alleles S1‐S23 and Sf. In addition, to improve discrimination between alleles of similar sizes, the same sets of primers but fluorescently labelled were used, and the products sized on an automated sequencer. These fluorescent primers were particularly informative in the case of the first intron, variation in the length of which has not been used previously for S genotyping in almond. Some reference alleles showed the same patterns with first and second intron primers, and others showed a microsatellite‐like trace. Subsequently, the S genotypes of 26 cultivars not genotyped previously and of four of uncertain genotype were determined. An allele described in Australian work as putative S10 was shown to be a ‘new’ allele and ascribed to S24 and evidence of five more ‘new’S alleles was found, for which the labels S25‐S29 are proposed. This PCR approach should be useful for genotyping in other Prunus crops.  相似文献   

14.
Use of self‐incompatibility (SI) as a pollination control method for Brassica napus hybrid production requires the development of a sufficient number of S‐alleles that are expressed consistently in a range of B. napus lines. Self‐incompatibility (SI) alleles have been transferred from Brassica oleracea and Brassica rapa into B. napus var. oleifera. An understanding of expression of these alleles in B. napus is essential for their commercial use. Four SI B. napus doubled haploids containing the B. oleracea S‐alleles S2, S5, S13 and S24 were crossed to three B. napus cultivars to measure the B. napus genetic background effect on S‐allele expression. A line x tester analysis indicated that the largest source of variation in the expression rate of SI was the S‐allele itself. The B. napus genotypes tested contained modifier gene(s), some that enhanced SI expression and others that inhibited SI expression. The B. napus Canadian cultivar ‘Westar’ generally had a negative effect on SI expression while the European cultivar ‘Topas’ had a positive effect on the B. oleracea S‐allele expression. The B. oleracea S‐allele S24 was very similar in expression to the B. rapa allele W1. The application of these results for the use of B. oleracea S‐alleles for hybrid production in B. napus is discussed.  相似文献   

15.
Apricot (Prunus armeniaca L.) shows gametophytic self-incompatibility controlled by a single locus with several allelic variants. An allele for self-compatibility (SC) and seven alleles for self-incompatibility (S1S7) were described previously. Our experiments were carried out to ascertain whether the number of allelic variants of apricot S-locus was indeed so small. Twenty-seven apricot accessions were analysed for stylar ribonucleases by non-equilibrium pH gradient electrofocusing (NEpHGE) to determine their S-genotype. To validate the results of electrofocusing, the applicability of the S-gene-specific consensus PCR primers designed from sweet cherry sequences was tested. NEpHGE revealed 12 bands associated with distinct S-alleles in newly genotyped cultivars. Cherry consensus primers amplified 11 alleles out from 16 ones, which indicated that these primers could also recognize most of the S-RNase sequences in apricot, and provided an efficient tool to confirm or reject NEpHGE results. By combining the protein and DNA-based methods, complete or partial S-genotyping was achieved for 23 apricot accessions and nine putatively new alleles (provisionally labelled S8S16) were found. Their identity needs to be confirmed by pollination tests or S-allele sequencing. This study provides evidence that similarly to other Prunus species, the S-locus of apricot is more variable than previously believed.  相似文献   

16.
Gliadin alleles were identified in 100 common wheat cultivars registered and/or grown in Spain during the last 40 years. A very high level of genetic polymorphism was found: in total, 103 allelic variants including one null‐allele were found at the six major Gli loci in the Spanish wheats studied. An average genetic diversity for these six loci was found to be higher (H=0.844) than in any group of wheat cultivars studied previously. Spanish wheats bred in Spain demonstrated even higher genetic diversity (H=0.868), probably because of the occurrence in this group of some landraces (local varieties) assumed to be strongly differentiated to fit local environments. The high level of genetic diversity of wheats grown in Spain was maintained by the introduction of distantly related wheat germplasm from different sources, especially from Italy and CIMMYT. A slight decrease of genetic diversity in recently registered cultivars might be caused by the excessive introduction of French wheats. Thirteen new alleles found in Spanish wheats were catalogued, including Gli‐D2w which encodes the first Gli‐D2‐controlledγ‐gliadin to be found.  相似文献   

17.
Prunus avium is primarily cultivated for its fruit, sweet cherries. However, it is also used to produce high‐quality timber. In a P. avium seed orchard, gametophytic self‐incompatibility is a restriction for free pollen flow and should be considered when establishing basic forest materials. In this study, S‐locus diversity and cross‐incompatibility of wild cherry individuals in clonal banks established for breeding for timber production were investigated. Wild cherry trees (140) with outstanding forest growth habit, collected in northern Spain, grafted and planted in two clonal banks, were genotyped at the S‐locus. The self‐incompatibility S‐locus genes, S‐RNase and SFB, were analysed by PCR. Twenty‐two S‐haplotypes, resulting in 72 different S‐genotypes, were identified. The genotypes were grouped into 33 incompatibility groups and 39 unique genotypes. This initial S‐locus analysis revealed large genetic diversity of wild cherry trees from the Spanish northern deciduous forest, and provides useful information for seed orchard design. Wild P. avium displays significantly more genetic diversity than what is detected in local cultivars, revealing a narrowing of genetic diversity during local domestication.  相似文献   

18.
Summary Protein stylar extracts of 16 cultivars of sweet cherry (Prunus avium), from the 10 different incompatibility groups to which incompatibility alleles have been assigned, were separated on acrylamide gels using isoelectric focusing (IEF) and were stained for ribonuclease activity. When two cultivars from the same incompatibility group were analyzed they gave identical zymograms and the cultivars of the 10 different incompatibility groups gave in all eight distinct zymograms. The ribonuclease polymorphism could be correlated with the reported S allele constitutions of the cultivars. Three ribonuclease bands were identified that each consistently corresponded to one of the six known incompatibility alleles (S 1, S2 and S 6), a fourth band apparently corresponded to S 3 and to the combination of S 4 and S 5, and a fifth band to S 4 and S 5 in other combinations. Thus, it seems that S alleles of cherry have ribonuclease activity and that IEF is useful for distinguishing S allele constitutions. The ribonuclease pattern of Summit, a cultivar of unknown incompatibility group, indicated its incompatibility genotype to be S 1S2, and this was confirmed by controlled pollination. The same band corresponded to S 4 and S 4', the mutant allele in self-compatible cultivars. IEF and ribonuclease staining promise to be useful tools for exploring the incompatibility relationships of cherry cultivars and perhaps of other self-incompatible Prunus crops.  相似文献   

19.
A top‐cross‐mating design among 29 S4 inbred lines and tester (cultivar ‘Dukat’) was carried out to study their breeding value in terms of general combining ability (GCA). The objectives of this study were to evaluate the acidity, soluble solids and dry matter contents in fruits of progeny F1 in comparison with S4 inbred lines as well as the cultivars (S0); identify strawberry genotypes with high value of GCA for use in cultivar development; and determine mid‐parent heterosis regarding S4 inbred lines and cultivated strawberry. The 2‐year observations showed statistically significant differences between tested genotypes in terms of the studied traits. The highest breeding value based on GCA was estimated for Chandler 123‐5 for soluble solids and dry matter content, and Kent 7‐6 for acidity. Estimated mid‐parent heterosis had positive and negative values. The highest heterosis in terms of extract and dry matter content (26.71% and 17.50%, respectively) occurred in the offspring Chandler 123‐5 × ‘Dukat’, but as regards acidity in hybrid Chandler 123‐22 with cv. ‘Dukat’. The study of genetic divergence by dendrograms may help to identify parents suitable for obtaining hybrids with higher heterosis effects.  相似文献   

20.
G. W. Wang    H. Y. Cai    Y. Xu    S. H. Yang    Y. Q. He 《Plant Breeding》2009,128(5):451-457
Wide‐compatibility varieties are a special class of rice germplasm that is able to produce fertile hybrids when crossed to either indica or japonica subspecies. Previous studies determined the f5 allele from ‘Dular’ (f5‐Du), f6 allele from ‘Dular’ (f6‐Du) and S5 allele from ‘02428’ (S5‐08) as neutral alleles conferring wide‐compatibility. However, the possible extent of the effect of the three neutral alleles has not been fully characterized because of the narrow range of the tester varieties used and the highly complex differentiation in Asian cultivated rice. In this study, we further developed the five near‐isogenic lines with higher recovery rates of the recurrent parent genome, and testcrossed to 14 japonica varieties, which have been widely used in rice breeding programmes in China. The results clearly revealed that all three neutral alleles exhibited substantial effects on spikelet fertility in most of the indicajaponica testcrosses, which indicated that these hybrid sterility loci have been extensively differentiated between indica and japonica varieties. The magnitudes of effects on spikelet fertility averaged over various crosses seem to be similar among the three neutral alleles, with f5‐Du, f6‐Du and S5‐08 alleles increasing spikelet fertility by 15.09%, 13.99% and 14.25%, respectively. The testcrosses involving f5‐Du allele generally showed much smaller variation in pollen fertility than others. The pyramiding lines with two neutral alleles showed a wider spectrum and a higher level of wide compatibility than others, whereas most of the increases in hybrid fertility couldn’t be simply explained by additive effects, suggesting the very complexity of wide compatibility and hybrid sterility. The indicajaponica hybrids involving restorer lines as one of their parents showed much higher pollen fertility (almost normal) and also higher spikelet fertility. The implications of the findings in rice breeding programmes are also discussed.  相似文献   

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