PCR markers for mutated <Emphasis Type="Italic">S</Emphasis>-haplotypes enable discrimination between self-incompatible and self-compatible sour cherry selections

Tetraploid sour cherry (Prunus cerasus L.) exhibits gametophytic self-incompatibility (GSI) whereby the specificity of self-pollen rejection is controlled by alleles of the stylar and pollen specificity genes, the S-RNase and SFB (S haplotype-specific F-box protein gene), respectively. As sour cherry selections can be either self-compatible (SC) or self-incompatible (SI), polyploidy per se does not result in SC. Instead, the genotype dependent loss of SI in sour cherry is due to the accumulation of non-functional S-haplotypes. The presence of two or more non-functional S-haplotypes within sour cherry 2x pollen renders that pollen SC. We previously determined that sour cherry has non-functional S-haplotypes for the S ₁ -, S ₆ - and S ₁₃ -haplotypes that are also present in diploid sweet cherry (P. avium L.). The mutations underlying these non-functional S-haplotypes have been determined to be structural alterations of either the S-RNase or SFB. Based on these structural alterations we designed derived cleaved amplified polymorphic sequence (dCAPS) markers and S-haplotype specific primer pairs that took advantage of either the length polymorphisms between S-haplotypes, differential S-haplotype sequences, or differential restriction enzyme cut sites. These primer pairs can discriminate among the mutant and wild-type S-haplotypes thereby enabling the identification of the S-haplotypes present in a sour cherry individual. This information can be used to determine whether the individual is either SC or SI. In a sour cherry breeding program, the ability to discriminate between SI and SC individuals at the seedling stage so that SI individuals can be discarded prior to field planting, dramatically increases the program’s efficiency and cost-effectiveness.     

Selbstfertile Diploide aus Polyploiden selbststeriler Petunien; Die Duplikation desS-Locus

Summary Crosses between self-incompatible triploidPetunia hybrida-plants and selfincompatible diploid ones produce self-compatible trisomics.Self-pollinations of the self-compatible trisomics give a great number of self-compatible diploid individuals which are constant self-compatible in further generations.It is supposed that self-compatibility is caused by a duplication of theS-locus. According to the results the duplication as well of identical as of not identicalS-Alleles must cause the break-down of the inhibition-reaction.Further it is supposed that the influence of the alleles in a duplicated locus both in pollen tubes and in styles is not sufficient to eliminate the inhibition-effect of normalS-genes which are presented in the reaction partner.

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Dissertation der Mathematisch-naturwissenschaftlichen Fakultät der Universität zu Köln.     

An S-Locus Independent Pollen Factor Confers Self-Compatibility in ‘Katy’ Apricot

Loss of pollen-S function in Prunus self-compatible cultivars has been mostly associated with deletions or insertions in the S-haplotype-specific F-box (SFB) genes. However, self-compatible pollen-part mutants defective for non-S-locus factors have also been found, for instance, in the apricot (Prunus armeniaca) cv. ‘Canino’. In the present study, we report the genetic and molecular analysis of another self-compatible apricot cv. termed ‘Katy’. S-genotype of ‘Katy’ was determined as S ₁ S ₂ and S-RNase PCR-typing of selfing and outcrossing populations from ‘Katy’ showed that pollen gametes bearing either the S ₁- or the S ₂-haplotype were able to overcome self-incompatibility (SI) barriers. Sequence analyses showed no SNP or indel affecting the SFB ₁ and SFB ₂ alleles from ‘Katy’ and, moreover, no evidence of pollen-S duplication was found. As a whole, the obtained results are compatible with the hypothesis that the loss-of-function of a S-locus unlinked factor gametophytically expressed in pollen (M’-locus) leads to SI breakdown in ‘Katy’. A mapping strategy based on segregation distortion loci mapped the M’-locus within an interval of 9.4 cM at the distal end of chr.3 corresponding to ∼1.29 Mb in the peach (Prunus persica) genome. Interestingly, pollen-part mutations (PPMs) causing self-compatibility (SC) in the apricot cvs. ‘Canino’ and ‘Katy’ are located within an overlapping region of ∼273 Kb in chr.3. No evidence is yet available to discern if they affect the same gene or not, but molecular markers seem to indicate that both cultivars are genetically unrelated suggesting that every PPM may have arisen independently. Further research will be necessary to reveal the precise nature of ‘Katy’ PPM, but fine-mapping already enables SC marker-assisted selection and paves the way for future positional cloning of the underlying gene.     

Inheritance of Hetero-Diploid Pollen S-Haplotype in Self-Compatible Tetraploid Chinese Cherry (Prunus pseudocerasus Lindl)

The breakdown of self-incompatibility, which could result from the accumulation of non-functional S-haplotypes or competitive interaction between two different functional S-haplotypes, has been studied extensively at the molecular level in tetraploid Rosaceae species. In this study, two tetraploid Chinese cherry (Prunus pseudocerasus) cultivars and one diploid sweet cherry (Prunus avium) cultivar were used to investigate the ploidy of pollen grains and inheritance of pollen-S alleles. Genetic analysis of the S-genotypes of two intercross-pollinated progenies showed that the pollen grains derived from Chinese cherry cultivars were hetero-diploid, and that the two S-haplotypes were made up of every combination of two of the four possible S-haplotypes. Moreover, the distributions of single S-haplotypes expressed in self- and intercross-pollinated progenies were in disequilibrium. The number of individuals of the two different S-haplotypes was unequal in two self-pollinated and two intercross-pollinated progenies. Notably, the number of individuals containing two different S-haplotypes (S₁- and S₅-, S₅- and S₈-, S₁- and S₄-haplotype) was larger than that of other individuals in the two self-pollinated progenies, indicating that some of these hetero-diploid pollen grains may have the capability to inactivate stylar S-RNase inside the pollen tube and grow better into the ovaries.     

Development of Self-Compatible B. rapa by RNAi-Mediated S Locus Gene Silencing

The self-incompatibility (SI) system is genetically controlled by a single polymorphic locus known as the S-locus in the Brassicaceae. Pollen rejection occurs when the stigma and pollen share the same S-haplotype. Recognition of S-haplotype specificity has recently been shown to involve at least two S-locus genes, S-receptor kinase (SRK) and S-locus protein 11 or S locus Cysteine-rich (SP11/SCR) protein. Here, we examined the function of S₆₀, one SP11/SCR allele of B. rapa cv. Osome, using a RNAi-mediated gene silencing approach. The transgenic RNAi lines were highly self-compatible, and this trait was stable in subsequent generations, even after crossing with other commercial lines. These findings also suggested that the resultant self-compatibility could be transferred to commercial cultivars with the desired performances in B. rapa.     

Genetic Features of the Spontaneous Self-Compatible Mutant, ‘Jin Zhui’ (Pyrus bretschneideri Rehd.)

‘Jin Zhui’ is a spontaneous self-compatible mutant of ‘Ya Li’ (Pyrus bretschneideri Rehd. S₂₁S₃₄), the latter displaying a typical S-RNase-based gametophytic self-incompatibility (GSI). The pollen-part mutation (PPM) of ‘Jin Zhui’ might be due to a natural mutation in the pollen-S gene (S₃₄ haplotype). However, the molecular mechanisms behind these phenotypic changes are still unclear. In this study, we identified five SLF (S-Locus F-box) genes in ‘Ya Li’, while no nucleotide differences were found in the SLF genes of ‘Jin Zhui’. Further genetic analysis by S-RNase PCR-typing of selfed progeny of ‘Jin Zhui’ and ‘Ya Li’ × ‘Jin Zhui’ progeny showed three progeny classes (S₂₁S₂₁, S₂₁S₃₄ and S₃₄S₃₄) as opposed to the two classes reported previously (S₂₁S₃₄ and S₃₄S₃₄), indicating that the pollen gametes of ‘Jin Zhui’, bearing either the S₂₁- or S₃₄-haplotype, were able to overcome self-incompatibility (SI) barriers. Moreover, no evidence of pollen-S duplication was found. These findings support the hypothesis that loss of function of S-locus unlinked PPM expressed in pollen leads to SI breakdown in ‘Jin Zhui’, rather than natural mutation in the pollen-S gene (S₃₄ haplotype). Furthermore, abnormal meiosis was observed in a number of pollen mother cells (PMCs) in ‘Jin Zhui’, but not in ‘Ya Li’. These and other interesting findings are discussed.     

Characterization of self-compatibility in sweet cherry varieties by crossing experiments and molecular genetic analysis

Self-compatibility is a major breeding objective in sweet cherry. The identification and characterization of new sources of self-compatibility will be useful for breeding and research purposes. In this work, self-compatibility of four local Spanish sweet cherry varieties was investigated by crossing experiments and molecular genetic analysis of two self-incompatibility loci. Crossing experiments included self- and cross-pollinations in the laboratory followed by microscopic observation of pollen tube growth and fruit set assay in the field. After crossing experiments, two accessions, ‘Son Miró’ and ‘Talegal Ahín’, were self-compatible while the other two were self-incompatible. Inheritance of S-locus and microsatellite EMPaS02 (linked to self-compatibility, Sc) were investigated in self-pollination progeny of both self-compatible genotypes. Results indicate that self-compatibility in ‘Talegal Ahín’ is similar to self-compatibility described in sweet cherry ‘Cristobalina’ and may be caused by the same mutation. That is a pollen part mutation not linked to the S-locus but linked to microsatellite EMPaS02 in cherry LG3. In ‘Son Miró’ self-compatibility seems more complex, affecting pollen and style function, and probably involving more than one mutation not described previously in sweet cherry. Together with ‘Cristobalina’, the newly described self-compatible varieties ‘Son Miró’ and ‘Talegal Ahín’ confirm the existence of unique self-compatible plant material in local germplasm from Spain that should be conserved and characterized for its use in breeding and research.     

On the origin of the widespread self-compatible allotetraploid Capsella bursa-pastoris (Brassicaceae)

Polyploidy, or whole-genome duplication, is a common speciation mechanism in plants. An important barrier to polyploid establishment is a lack of compatible mates. Because self-compatibility alleviates this problem, it has long been hypothesized that there should be an association between polyploidy and self-compatibility (SC), but empirical support for this prediction is mixed. Here, we investigate whether the molecular makeup of the Brassicaceae self-incompatibility (SI) system, and specifically dominance relationships among S-haplotypes mediated by small RNAs, could facilitate loss of SI in allopolyploid crucifers. We focus on the allotetraploid species Capsella bursa-pastoris, which formed ~300 kya by hybridization and whole-genome duplication involving progenitors from the lineages of Capsella orientalis and Capsella grandiflora. We conduct targeted long-read sequencing to assemble and analyze eight full-length S-locus haplotypes, representing both homeologous subgenomes of C. bursa-pastoris. We further analyze small RNA (sRNA) sequencing data from flower buds to identify candidate dominance modifiers. We find that C. orientalis-derived S-haplotypes of C. bursa-pastoris harbor truncated versions of the male SI specificity gene SCR and express a conserved sRNA-based candidate dominance modifier with a target in the C. grandiflora-derived S-haplotype. These results suggest that pollen-level dominance may have facilitated loss of SI in C. bursa-pastoris. Finally, we demonstrate that spontaneous somatic tetraploidization after a wide cross between C. orientalis and C. grandiflora can result in production of self-compatible tetraploid offspring. We discuss the implications of this finding on the mode of formation of this widespread weed.Subject terms: Evolution, Polyploidy in plants, Plant evolution, Haplotypes     

Breakdown of self-incompatibility in tetraploid Lycopersicon peruvianum: inheritance and expression of S-related proteins

 Lycopersicon peruvianum displays gametophytic self-incompatibility (GSI). We have isolated self-compatible (SC) tetraploids of L. peruvianum from tissue-cultured leaves and have explored the expression and inheritance of their S-related proteins. The Srelated protein profiles of styles of SC tetraploids were indistinguishable from the diploid self-incompatible (SI) explant source based on SDS-PAGE. All progeny obtained from self-fertilization of two tetraploids were SC. Cloned cDNA sequences of the S-related proteins were used to determine the inheritance of this locus in these progeny through Southern hybridization. The allelic ratio, as determined from the intensity of DNA restriction fragments, was consistent with the predicted ratio if only pollen bearing two different alleles was successful in achieving fertilization. All progeny obtained had at least one copy of each allele, and individuals fully homozygous for either allele were not found, indicating that pollen grains bearing two identical alleles were inhibited. In addition, the level of expression of the S-related proteins in the progeny correlated with the allelic dosage at the DNA level. We demonstrate that the observed self-compatibility in the tetraploids was not caused by an alteration in the expression of S-related proteins. Received: 11 September 1996 / Accepted: 21 March 1997     

Variability of the self-incompatibility reaction in Brassica oleracea L. with S 15 haplotype

Self-incompatibility (SI) is thought to have played a key role in the evolution of species as it promotes their outcrossing through the recognition and rejection of self-pollen grains. In most species, SI is under the control of a complex, multiallelic S-locus. The recognition system is associated with quantitative variations of the strength of the SI reaction; the origin of these variations is still not elucidated. To define the genetic regulations involved, we studied the variability of the SI response in homozygous S ₁₅ S ₁₅ plants in cauliflower. These plants were obtained from a self-progeny of a self-compatible (SC) plant heterozygous for S ₁₅ , which was generated after five selfing generations from one strongly self-incompatible initial plant. We found a continuous phenotypic variation for SI response in the offspring plants homozygous for the S ₁₅ haplotype, from the strict SI reaction to self-compatibility, with a great proportion of the plants being partially self-compatible (PSC). Decrease in SI levels was also observed during the life of the flower. The number of pollen tubes passing through the stigma barrier was higher when counted 3 or 5 days after pollination than one day after pollination. Analysis of the expression of the two key genes regulating self-pollen recognition in cauliflower, the S-locus receptor kinase (SRK) and S-locus cysteine-rich (SCR/SP11) genes, revealed that self-compatibility or PSC was associated with decreased SRK or SCR/SP11 expression. Our work shows the particularly high level of phenotypic plasticity of the SI response associated with certain S-haplotypes in cauliflower.     

Genetic and molecular analysis in Cristobalina sweet cherry,a spontaneous self-compatible mutant

Self-compatibility in a naturally self-incompatible species like sweet cherry is a highly interesting trait for breeding purposes and a powerful tool with which to investigate the basis of the self-incompatible reaction in gametophytic systems. However, natural self-compatibility in sweet cherry is a very rare phenomenon. Cristobalina is a local Spanish sweet cherry cultivar that has proven to be spontaneously self-compatible. In this work, the nature of the self-compatibility in Cristobalina has been studied using genetic and molecular approaches. Pollination studies and microscopic observations of pollen tube growth were carried out to confirm the self-compatible character and the results obtained indicate that self-compatibility is caused by a failure of the pollen and not the style factor. Polymerase chain reaction (PCR) analysis of progenies derived from Cristobalina revealed that self-compatibility in this genotype is not related uniquely to one of the two pollen S alleles, but that pollen grains carrying either of the two haplotypes can overcome the incompatibility barrier. Moreover, PCR analysis and microscopic observation of pollen tube growth in progeny derived from Cristobalina also confirmed that the self-compatible descendants can carry either of the two S haplotypes of their progenitor. Isolation and sequencing of the style S-RNases and pollen SFBs revealed that the DNA sequences of these factors are the same as those described in other self-incompatible sweet cherry cultivars with the same S alleles. Possible mechanisms to explain self-compatibility in Cristobalina are discussed.     

Self-compatibility of two apricot selections is associated with two pollen-part mutations of different nature

Loss of pollen-S function in Prunus self-compatible mutants has recently been associated with deletions or insertions in S-haplotype-specific F-box (SFB) genes. We have studied two self-compatible cultivars of apricot (Prunus armeniaca), Currot (S(C)S(C)) and Canino (S(2)S(C)), sharing the naturally occurring self-compatible (S(C))-haplotype. Sequence analysis showed that whereas the S(C)-RNase is unaltered, a 358-bp insertion is found in the SFB(C) gene, resulting in the expression of a truncated protein. The alteration of this gene is associated with self-incompatibility (SI) breakdown, supporting previous evidence that points to SFB being the pollen-S gene of the Prunus SI S-locus. On the other hand, PCR analysis of progenies derived from Canino showed that pollen grains carrying the S(2)-haplotype were also able to overcome the incompatibility barrier. However, alterations in the SFB(2) gene or evidence of pollen-S duplications were not detected. A new class of F-box genes encoding a previously uncharacterized protein with high sequence similarity (approximately 62%) to Prunus SFB proteins was identified in this work, but the available data rules them out of producing S-heteroallelic pollen and thus the cause of the pollen-part mutation. These results suggest that cv Canino has an additional mutation, not linked to the S-locus, which causes a loss of pollen-S activity when present in pollen. As a whole, these findings support the proposal that the S-locus products besides other S-locus independent factors are required for gametophytic SI in Prunus.     

Duplication of the <Emphasis Type="Italic">S</Emphasis>-locus F-box gene is associated with breakdown of pollen function in an <Emphasis Type="Italic">S</Emphasis>-haplotype identified in a natural population of self-incompatible <Emphasis Type="Italic">Petunia axillaris</Emphasis>

We previously identified both self-incompatible and self-compatible plants in a natural population of self-incompatible Petunia axillaris subsp. axillaris, and found that all the self-compatible plants studied carried either S_C1- or S_C2-haplotype. Genetic crosses showed that S_C2 was identical to S₁₇ identified from another natural population of P. axillaris, except that its pollen function was defective, and that the pollen-part mutation in S_C2 was tightly linked to the S-locus. Recent identification of the S-locus F-box gene (SLF) as the gene that controls pollen specificity in S-RNase-based self-incompatibility has prompted us to examine the molecular basis of this pollen-part mutation. We cloned and sequenced the S₁₇-allele of SLF of P.axillaris, named PaSLF₁₇, and found that S_C2 S_C2 plants contained extra restriction fragments that hybridized to PaSLF₁₇ in addition to all of those observed in S₁₇ S₁₇ plants. Moreover, these additional fragments co-segregated with S_C2. We used the S_C2-specific restriction fragments as templates to clone an allele of PaSLF by PCR. To determine the identity of this allele, named PaSLF_x, primers based on its sequence were used to amplify PaSLFalleles from genomic DNA of 40 S-homozygotes of P. axillaris, S₁ S₁ through S₄₀ S₄₀. Sequence comparison revealed that PaSLF_x was completely identical with PaSLF₁₉ obtained from S₁₉ S₁₉. We conclude that the S-locus of S_C2 contained both S₁₇-allele and the duplicated S₁₉-allele of PaSLF. S_C2 is the first naturally occurring pollen-part mutation of a solanaceous species that was shown to be associated with duplication of the pollen S. This finding lends support to the proposal, based on studies of irradiation-generated pollen-part mutants of solanaceous species, that duplication, but not deletion, of the pollen S, causes breakdown of pollen function.     

Pollen-expressed F-box gene family and mechanism of S-RNase-based gametophytic self-incompatibility (GSI) in Rosaceae

Many species of Rosaceae, Solanaceae, and Plantaginaceae exhibit S-RNase-based self-incompatibility (SI) in which pistil-part specificity is controlled by S locus-encoded ribonuclease (S-RNase). Although recent findings revealed that S locus-encoded F-box protein, SLF/SFB, determines pollen-part specificity, how these pistil- and pollen-part S locus products interact in vivo and elicit the SI reaction is largely unclear. Furthermore, genetic studies suggested that pollen S function can differ among species. In Solanaceae and the rosaceous subfamily Maloideae (e.g., apple and pear), the coexistence of two different pollen S alleles in a pollen breaks down SI of the pollen, a phenomenon known as competitive interaction. However, competitive interaction seems not to occur in the subfamily Prunoideae (e.g., cherry and almond) of Rosaceae. Furthermore, the effect of the deletion of pollen S seems to vary among taxa. This review focuses on the potential differences in pollen-part function between subfamilies of Rosaceae, Maloideae, and Prunoideae, and discusses implications for the mechanistic divergence of the S-RNase-based SI.     

Self-incompatibility is codominant in intraspecific hybrids of self-compatible and self-incompatible Lycopersicon peruvianum and L. hirsutum based on protein and DNA marker analysis

Self-compatibility was investigated separately in two species of tomato, Lycopersicon peruvianum and L. hirsutum. The codominant expression of self-compatibility (SC)/self incompatibility (SI) was established using intraspecific hybrids of SC and SI hybrids. In SC L. peruvianum, a major stylar protein of approximately 29 kDa cosegregates with self-compatibility in the progeny of SC/SI hybrids. The SC/SI hybrids are self-fertile, but only partially so, since the SI allele present in the hybrids is capable of eliminating certain genotypes in the resultant progeny. In L. hirsutum, the majority of hybrids between one accession of SI L. hirsutum f. hirsutum and one of SC L. hirsutum f. glabratum are self-fertile. Analysis of the progeny revealed that the SC and SI alleles are codominant in this species as well. A protein product for the SC allele is not obvious in style extracts of L. hirsutum f. glabratum. Segregating progeny from SC/SI hybrids of L. hirsutum were used to map the S locus against five RFLP markers on chromosome 1, and estimated map distances are given. In addition, evidence is presented that indicates that one of the DNA markers, CD15, is duplicated in L. hirsutum f. glabratum, and the duplication is not linked to the S locus.     

Genetic features of a pollen-part mutation suggest an inhibitory role for the <Emphasis Type="Italic">Antirrhinum</Emphasis> pollen self-incompatibility determinant

Self-incompatibility (SI), an important barrier to inbreeding in flowering plants, is controlled in many species by a single polymorphic S-locus. In the Solanaceae, two tightly linked S-locus genes, S-RNase and SLF (S-locus F-box)/SFB (S-haplotype-specific F-box), control SI expression in pistil and pollen, respectively. The pollen S-determinant appears to function to inhibit all but self S-RNase in the Solanaceae, but its genetic function in the closely-related Plantaginaceae remains equivocal. We have employed transposon mutagenesis in a member of the Plantaginaceae (Antirrhinum) to generate a pollen-part SI-breakdown mutant Pma1 (Pollen-part mutation in Antirrhinum1). Molecular genetic analyses showed that an extra telocentric chromosome containing AhSLF-S ₁ is present in its self-compatible but not in its SI progeny. Furthermore, analysis of the effects of selection revealed positive selection acting on both SLFs and SFBs, but with a stronger purifying selection on SLFs. Taken together, our results suggest an inhibitor role of the pollen S in the Plantaginaceae (as represented by Antirrhinum), similar to that found in the Solanaceae. The implication of these findings is discussed in the context of S-locus evolution in flowering plants. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Yongbiao Xue, Yijing Zhang, and Qiuying Yang contributed equally to this work.     

Breakdown of self-incompatibility in a natural population of Petunia axillaris caused by loss of pollen function

Although Petunia axillaris subsp. axillaris is described as a self-incompatible taxon, some of the natural populations we have identified in Uruguay are composed of both self-incompatible and self-compatible plants. Here, we studied the self-incompatibility (SI) behavior of 50 plants derived from such a mixed population, designated U83, and examined the cause of the breakdown of SI. Thirteen plants were found to be self-incompatible, and the other 37 were found to be self-compatible. A total of 14 S-haplotypes were represented in these 50 plants, including two that we had previously identified from another mixed population, designated U1. All the 37 self-compatible plants carried either an S(C1)- or an S(C2)-haplotype. S(C1)S(C1) and S(C2)S(C2) homozygotes were generated by self-pollination of two of the self-compatible plants, and they were reciprocally crossed with 40 self-incompatible S-homozygotes (S(1)S(1) through S(40)S(40)) generated from plants identified from three mixed populations, including U83. The S(C1)S(C1) homozygote was reciprocally compatible with all the genotypes examined. The S(C2)S(C2) homozygote accepted pollen from all but the S(17)S(17) homozygote (identified from the U1 population), but the S(17)S(17) homozygote accepted pollen from the S(C2)S(C2) homozygote. cDNAs encoding S(C2)- and S(17)-RNases were cloned and sequenced, and their nucleotide sequences were completely identical. Analysis of bud-selfed progeny of heterozygotes carrying S(C1) or S(C2) showed that the SI behavior of S(C1) and S(C2) was identical to that of S(C1) and S(C2) homozygotes, respectively. All these results taken together suggested that the S(C2)-haplotype was a mutant form of the S(17)-haplotype, with the defect lying in the pollen function. The possible nature of the mutation is discussed.     

Two Different Prunus SFB Alleles Have the Same Function in the Self-incompatibility Reaction

Many species in the families of Rosaceae, Solanaceae, and Scrophulariaceae exhibit gametophytic self-incompatibility, a phenomenon controlled by two polymorphic genes at the S-locus, style-S (S-RNase) and pollen-S (SFB). Sequences of both genes show high levels of diversity, characteristic of genes involved in recognition of self-incompatibility systems in plants. In this study, S ₂₄ -RNase and SFB ₂₄ alleles were cloned from Prunus armeniaca cv. Chuanzhihong (Chinese apricot). Sequence comparisons of deduced amino acid sequences revealed that the P. armeniaca S ₂₄ -haplotype has different SFB alleles, but shares a single S-RNase allele with P. armeniaca S ₄ -haplotype. Moreover, P. armeniaca S ₂₄ -RNase haplotype has a single and three different alleles with S ₁ -RNase of P. tenella (dwarf almond) and S ₁ -RNase of P. mira (smooth pit peach), respectively. The functionalities of SFB ₂₄ and SFB ₄ have been evaluated by pollen tube growth and controlled field tests of P. tenella and P. mira. Genetic analysis of the two intercrosses showed that progenies segregated 1:1 into two S-genotype classes, which is consistent with the expected ratio for semi-compatibility. These findings imply that the allelic function of the S ₂₄ -haplotype is identical to that of the S ₄ -haplotype in a self-incompatibility reaction. Thus, these two Prunus S-haplotypes are in fact two neutral variants of the same S-haplotype. The evolution of the S-allele is also discussed in terms of both functions and differences between S ₂₄ - and S ₄ -haplotypes in Prunus.     

Protein expression of a self-compatible allele from Lycopersicon peruvianum: introgression and behavior in a self-incompatible background

Lycopersicon peruvianum (wild tomato) is a gametophytic self-incompatible (SI) species. One natural population has been shown to harbor a self-compatible (SC) allele. A stylar protein associated with the self-compatibility allele has been elucidated using SDS-PAGE. The temporal and spatial expression of this protein is presented and compared with protein expression of two SI alleles. Hybrids containing the SC and SI alleles were used in a backcrossing program to introgress the SC allele into SI backgrounds in six independent lines. Controlled pollinations and SDS-PAGE were used to identify and select classes of progeny. After four backcross generations (approximately 97% recovery of the SI backgrounds) the SC allele still confers self-fertility in lines that contain this allele, providing evidence that the mutation to SC occurred at the S-locus and that the associated protein is likely responsible.