首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献，搜索用时 187 毫秒
1.
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   

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.
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   

4.
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   

5.
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   

6.
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   

7.
A series of PCR methods were used to detect S-RNase alleles and SFB alleles and to determine S-genotypes in 25 accessions of myrobalan (Prunus cerasifera L.). Firstly, primers flanking the polymorphic second intron were used to identify S-RNases in agarose gels. These primers amplified one or two bands per accession in 25 accessions. Then consensus primers were designed for amplifying the polymorphic first intron, unique to Prunus S-RNases, for automated fluorescent detection. Each accession produced one or two peaks. New primers were then developed to amplify the intron in the SFB gene, for detection by fluorescence. Cross-referencing PCR bands and peaks indicated 15 S-alleles were present in the 25 accessions. Cloning, sequencing and comparison with published data indicated that the amplified products were S-RNase alleles. Sequence information was used to design primers specific for each S-RNase. Full and consistent S-genotypes were obtained by cross-comparing PCR data for 23 of the 25 accessions, and two accessions appeared to have a single allele. Pollen-tube microscopy indicated function of some but not all of the S-alleles sequenced.  相似文献   

8.
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   

9.
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   

10.
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 (S1–S7) 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 S8–S16) 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   

11.
Sexual self-incompatibility in wild diploid potato species is controlled by a single multiallelic S-locus encoding a polymorphic stylar ribonuclease (S-RNase) that is responsible for the female function in pollen–pistil recognition. In this study, an approach using PCR-based markers were originally developed to amplify the S-RNase alleles in Solanum chacoense. Subsequently, to investigate their general applicability in Solanum, this molecular approach was successfully tested on S. spegazzinii and S. kurtzianum. Application of PCR-SSCP approach revealed potentially new S-RNase alleles in the three species, demonstrating for the first time the existence of S-RNase genetic variability within and between populations of wild diploid potato species. Species-specific SSCP markers that may be successfully used in gene flow studies was also detected in this investigation.  相似文献   

12.
Identification of S-alleles in almond using multiplex PCR   总被引：1，自引：0，他引：1  
The S-genotypes of eight almond (Prunus dulcis Miller (D.A. Webb)) cultivars from different geographical origins and of nine new selections from the CEBAS-CSIC (Murcia, Spain) breeding program were determined using single and multiplex PCR with different sets of specific oligonucleotide primers. The results of PCR using the AS1II- and AmyC5R-specific primers showed amplification in a single reaction of 10 different self-incompatibility alleles and of the self-compatibility allele S f. However, the amplified fragments of the S f allele were of similar sizes to those amplified from the S 3 self-incompatibility allele. For this reason, a specific PCR primer CEBASf was designed from the intron sequence of S f. A multiplex-PCR reaction using the AS1II, CEBASf and AmyC5R primers permitted unequivocal identification of the 10 self-incompatibility alleles and of the self-compatibility allele. Multiplex PCR opens the possibility to identify new S-alleles using different sets of primers. The applications of these PCR markers in the almond-breeding programs are discussed.  相似文献   

13.
In previous work the existence of proteins with RNase activity associated with S alleles in apricot was demonstrated. These proteins were inherited as described previously for the inheritance of self‐compatibility in this species. In this study, new cultivars have been genotyped for self‐compatibility using this method and it has been demonstrated that in all self‐compatible cultivars examined, the self‐compatibility allele is the same and is associated with an RNase with high activity. Homozygous self‐compatible individuals have been detected among established cultivars as well as among seedlings following breeding activity. This germplasm is of great value within the breeding programme because only self‐compatible seedlings will be produced. The number of S alleles in apricot appears to be low and only eight different alleles have been found in the large number of different cultivars screened. Furthermore, there are alleles present in the Spanish population that are also found in the genetic pool of North American cultivars. The screening of a progeny from the cross between the American cultivar ‘Goldrich’ and the Spanish cultivar ‘Pepito’ demonstrated the existence of the common allele S2 (detected previously by examining RNases), which was confirmed by the segregation of self‐compatibility in the progeny.  相似文献   

14.
T. Sonneveld    T. P. Robbins    K. R. Tobutt 《Plant Breeding》2006,125(3):305-307
A novel polymerase chain reaction (PCR) approach to determine and confirm the self‐incompatibility (S) genotype of cherries is reported. The method involves PCR amplification with a new pair of consensus primers that immediately flank the first intron of cherry S‐RNases, one of which is fluorescently labelled. Fluorescent amplification products range from 234 to c. 460 bp and can be sized accurately on an automated sequencer. Thirteen S alleles reported in sweet cherry can be distinguished, except for S2 and S7, which have an amplification product of exactly the same size. S13, which is also amplified, gives a microsatellite‐like trace which shows minor intra‐allelic length variation. This method gives fast and accurate results and should be especially useful for medium/high‐throughput genotyping of wild and cultivated cherries.  相似文献   

15.
‘Marcona’ and ‘Desmayo Largueta’ are the most widely cultivated almonds in Spain, representing around 27% of the total production and 34% of the entire almond growing surface. The excellent quality of their kernels makes them highly appreciated and demanded worldwide. However, due to their self-incompatibility, they should be grown along with cross-compatible cultivars, whose lower kernel quality often reduces the economic benefits of the plantation. In addition, although they are cross-compatible, are not good candidates to share the orchard since ‘Desmayo Largueta’ usually flowers earlier than ‘Marcona’. Therefore, to optimize orchard yield, genotypes with overlapping flowering times, cross-compatible with these cultivars and of similar fruit and ripening characteristics are desirable. In order to find suitable pollinators of these cultivars, five ‘Marcona’ and four ‘Desmayo Largueta’-type selections from “Instituto Técnico Agronómico Provincial (ITAP) de Albacete” (Spain) were characterized for flowering time and for self and cross-incompatibility. The results obtained showed that the nine ITAP selections were self-incompatible, and that three and one were promising candidates as pollinators of ‘Marcona’ and ‘Desmayo Largueta’, respectively. The S-haplotypes of two ITAP selections were characterized by cloning and sequencing their pistil S (S-RNase) and pollen S (SFB) genes. The results also showed that S f RNase does not have a mutated histidine in C2 region, and detected differences with other previously published sequences for S 23 RNase and SFB 23 allele. Moreover, the sequence for almond SFB 27 allele is reported here for the first time.  相似文献   

16.
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   

17.
Radish (Raphanus sativus L.) is a typical cross‐pollinated crop that exhibits obvious heterosis. Self‐incompatibility is an important character for F1 hybrid breeding of radish. Knowledge of the S haplotypes of breeding lines is very important for breeders to avoid cross‐incompatibility of the parental lines. In the present study, the S haplotypes of 63 radish inbred lines, which were independently cultivated by our research group, were identified by PCR amplification, sequencing and BLAST analyses of the SRK and SLG genes. Finally, fifty‐four inbred lines were classified into 15 class I S haplotypes, including three new types, RsS‐38, RsS‐39 and RsS‐40. Additionally, three class II S haplotypes were identified in nine radish inbred lines. Partial SRK or SLG sequences were completed, such as RsS‐11 Lim (SRK‐S), RsS‐26 (SRK‐K), RsS‐5 Lim (SRK‐K and SRK‐S) and RsS‐9 (SRK‐K). The identified S haplotypes were verified with a cross‐pollination test, and RsS‐9 has weaker self‐incompatibility than other S haplotypes. These information will not only contribute to the production of hybrid seeds but also to the development of new self‐compatible inbred lines, which were advantageous of the production of maintain line and male line in CMS breeding system.  相似文献   

18.
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   

19.
S. Mohring    V. Horstmann  E. Esch 《Plant Breeding》2005,124(2):105-110
Using primers annealing to S locus sequences the cleaved amplified polymorphic sequences (CAPS) method was applied to develop a marker and to characterize different alleles at the self‐incompatibility locus in Brassica napus. A segregating F2 population from a cross of a self‐incompatible (SI) and a self‐compatible parent, as well as seven SI lines representing four different S alleles were used. Several primers specific to the S locus in B. oleracea and B. campestris, chosen from the literature, allow polymerase chain reaction (PCR) amplification of genomic DNA. However, only one primer pair amplified a single specific and reproducible PCR fragment of the expected length in B. napus. Digestion with restriction endonucleases revealed polymorphisms for two CAPS markers absolutely linked to the S locus. Using the codominant marker efMboI it was possible to discriminate all three F2 genotypes. With this marker and an additional marker using another primer pair it was possible to distinguish between three of the four different S alleles and five of the seven SI lines, respectively.  相似文献   

20.
Self‐incompatibility in Brassicaceae plants is sporophytically controlled by a single multi‐allelic locus (S locus), which contains at least three highly polymorphic genes expressed in the stigma (SLG and SRK) and in the pollen (SCR/SP11). Using polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) analysis with SXG‐specific primer pairs, the S haplotypes of F1 hybrid and open‐pollinated commercial cultivars of Brassica rapa were identified. The number of S haplotypes detected in the F1 hybrid cultivars of Chinese cabbage, komatsuna, pak‐choi, turnip, open‐pollinated cultivars of Chinese cabbage and turnip were 9, 9, 4, 11, 13 and 12, respectively. Nine of them had different PCR‐RFLP profiles from those of the S‐tester lines that determined the SLG sequences. Four SLG sequences in the F1 hybrid cultivars were determined and named S53, S54, S55 and S56, respectively. It is demonstrated that the PCR‐RFLP analysis using specific primer pairs of SLG and SRK is useful for identification of the S haplotypes, in both, S homozygous and S heterozygous plants of B. rapa. The possibility of using this method routinely in breeding programmes, and in the evaluation of F1 hybrid seed purity, is discussed.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号