The SKP1 gene is an important component of the SCF(SKP1-Cullin1-F-box)complex and serves as a bridge connecting the F-box and Cullin1genes(F-box-SKP1-Cullin1).The pattern of S-RNase being ubiquitously labelled by the ...The SKP1 gene is an important component of the SCF(SKP1-Cullin1-F-box)complex and serves as a bridge connecting the F-box and Cullin1genes(F-box-SKP1-Cullin1).The pattern of S-RNase being ubiquitously labelled by the SCF complex and degraded by the 26S protease accounts for the bulk of the available self-incompatibility studies.In this study,15 ClSKP1s from the‘Xiangshui'lemon genome and ubiquitome exist in the same SKP1 conserved domain(CD)as SKP1s in other species.The q PCR results showed that SKP1-6 and SKP1-14 have tissue expression patterns specific for expression in pollen.In addition,SKP1-6 and SKP1-14 in the stigma,style and ovary were significantly upregulated after self-pollination compared to those after cross-pollination.A subcellular location showed that SKP1-6 and SKP1-14 were located in the nucleus.In addition,yeast two-hybrid(Y2H)assays,bimolecular fluorescence complementation(BiFC)and luciferase complementation imaging(LCI)assays showed that SKP1-6 interacted with F-box1,F-box33,F-box34,F-box17,F-box19,Cullin1-2 and 26S proteasome subunit 4 homolog A(26S PS4HA).SKP1-14 interacted with F-box17,F-box19,F-box35,Cullin1-2 and 26S PS4HA.The interaction of Cullin1-2 and the F-box with SKP1 as a bridge was verified by a yeast three-hybrid experiment.The ability of S3-RNase to inhibit pollen and pollen tube growth and development was assessed using in vitro pollen co-culture experiments with recombinant S3-RNase proteins.Overall,this study provides important experimental evidence and theoretical basis for understanding the mechanism of self-incompatibility in plants by revealing the key role of the SCF complex in‘Xiangshui'lemon,which is bridged by ClSKP1-6,in self-incompatibility.The results of this study are of great significance for the future indepth exploration of the molecular mechanism of the SCF complex and its wide application in the self-incompatibility of plants.展开更多
Re-domestication of diploid potato(Solanum tuberosum)into a seed crop is an innovative breeding method to accelerate genetic improvement.Seed propagation would allow hybrid production and mix superior alleles.However,...Re-domestication of diploid potato(Solanum tuberosum)into a seed crop is an innovative breeding method to accelerate genetic improvement.Seed propagation would allow hybrid production and mix superior alleles.However,almost all diploid potatoes in nature are self-incompatible(SI).Gametophytic self-incompatible(GSI)is a widespread SI in Solanaceae and is controlled by the S locus that contains a ribonuclease(S-RNase)and multiple F-box(SLFs);however,the genetic diversity of the S locus in potato is unclear.This study identified 21 S-RNase alleles involved in SI from 194 diploid potato accessions by large-scale transcriptome sequencing.The levels of amino acid similarity among different S-RNase proteins varied from 31.3 to 95.8%.S2 allele is the most widespread in 194 diploid potatoes and is mainly distributed in the S.tuberosum Group Phureja.Based on genomic annotation and expression analysis,we identified 12 potential functional SI male-determinant genes,S-locus F-box(SLFs),encoding F-box proteins in the S2 locus on a genomic region of approximately 13 Mb.Comparative genomics analysis showed that eight SLF genes are relatively conserved among four homozygous S locus.The Ka and Ks analysis suggested that S-RNase and intra-haplotypic SLF genes have co-evolved.These findings help select suitable pollinators,combine more hybrid combinations,and fully use heterosis to accelerate diploid potato breeding.展开更多
In plants,heteromorphic self-incompatibility(HetSI)is a strategy for avoiding self-pollination and promoting outcrossing,and during this process,numerous protein-protein interaction events occur between the pistil and...In plants,heteromorphic self-incompatibility(HetSI)is a strategy for avoiding self-pollination and promoting outcrossing,and during this process,numerous protein-protein interaction events occur between the pistil and pollen.Previous studies in Primula and Fagopyrum that focused on HetSI systems have provided interesting insights;however,the molecular mechanism underlying HetSI remains largely unknown.In this study,we profiled the proteome of Plumbago auriculata stigmas before and after self-incompatible(SI)and self-compatible(SC)pollination.Comparative analyses were conducted by 4D-DIA(Four-dimensional data independent acquisition),a promising technology that increases the sensitivity and reduces the spectral complexity of proteomic analysis by adding a fourth dimension,ion mobility.The results revealed 33387 peptides and 5311 proteins in all samples.The pathways in which the differentially expressed proteins(DEPs)identified in the P×P(Pin style self-pollinated with pin pollen)vs.PS(Pin style)and T×T(Thrum style self-pollinated with thrum pollen)vs.TS(Thrum style)comparisons were significantly enriched were biosynthesis of secondary metabolites and pentose and glucuronate interconversions.In the P×T(Pin style cross-pollinated with thrum pollen)vs.PS and T×P(Thrum style cross-pollinated with pin pollen)vs.TS comparison,the top three pathways were biosynthesis of secondary metabolites,pentose and glucuronate interconversions,and phenylpropanoid biosynthesis.The phenylpropanoid biosynthesis,cutin,suberine and wax biosynthesis,and flavonoid biosynthesis pathways were enriched in the P×T vs.P×P comparison,and starch and sucrose metabolism,glycerophospholipid metabolism,and alpha-linolenic acid metabolism were abundant in the T×T vs.T×P comparison.The enriched pathways between PS and TS were the biosynthesis of secondary metabolites,phenylpropanoid biosynthesis,and pentose and glucuronate interconversion.Self-incompatibility protein S1(SI S1),Mitogen-activated protein kinase 3/4(MPK3/4),Mitogen-activated protein kinase kinase 2/3(M2K2/3),Exocyst complex component EXO70A1(E70A1)and Thioredoxin H1/2(TRXH1/2)were found to be HetSI-related candidates,and O-fucosyltransferase 23(OFT23),3-ketoacyl-CoA synthase 6(KCS6),Receptor-like protein kinase FERONIA(FERON),Fimbrin-5(FIMB5),Pollen-specific leucine-rich repeat extensin-like protein 4(PLRX4),Transcription initiation factor IIB-2(TF2B2)and Pectinesterase 1(AL11A),etc.,were identified as other regulatory transducers.These findings combined with our morphological and reactive oxygen species(ROS)intensity analyses indicate that P.auriculata has typical dry-stigmas and that the HetSI mechanism might differ between the pin and thrum.SI S1 might be the key factor in HetSI,and ROS are overexpressed during SC pollination to rapidly activate the mitogen-activated protein kinase(MAPK)-mediated phosphorylation of E70A1 to maintain stigma receptivity in plants with HetSI.展开更多
The self-incompatibility ( S) loci from the Solanaceae, Rosaceae and Scrophulariaceae encode a class of ribonucleases, known as S RNases, which have been shown to control the pistil expression of self-incompatible rea...The self-incompatibility ( S) loci from the Solanaceae, Rosaceae and Scrophulariaceae encode a class of ribonucleases, known as S RNases, which have been shown to control the pistil expression of self-incompatible reaction. In the former two families, the S loci have been shown to be located near centromere. However, the chromosomal location of the S locus in Antirrhinum, a species of the Scrophulariaceae, is not known. To determine its chromosomal location and genomic organization, an S-2 RNase gene and its corresponding 63 kb BAC clone were separately used for fluorescence in situ hybridization (FISH) of mitotic metaphase chromosomes of a self-incompatible Antirrhinum line Of S2S5. The results showed that the S-2 RNase detected a doublet signal near the centromere of the smallest chromosome (2n = 16). Two separate doublet signals of the tested BAC sequence were shown on both sides of the centromeres of all eight pairs of the chromosomes, suggesting that the Antirrhinum S locus is located in a pericentromeric region. Furthermore, a retrotransposon, named RIS1 (retrotransposon in the S locus), which has not been identified yet in. Antirrhinum, was found next to S-2 RNase. Taken together, the centromeric location of the S locus from the three S-RNase-based self-incompatible families provides a further support on a common origin of their evolution as well as suppressed recombination.展开更多
In flowering plants, self-incompatibility （SI） serves as an important intraspecific reproductive barrier to promote outbreeding. In species from the Solanaceae, Plantaginaceae and Rosaceae, S-RNase and SLF （S-locus...In flowering plants, self-incompatibility （SI） serves as an important intraspecific reproductive barrier to promote outbreeding. In species from the Solanaceae, Plantaginaceae and Rosaceae, S-RNase and SLF （S-locus F-box） proteins have been shown to control the female and male specificity of SI, respectively. However, little is known about structure features of the SLF protein apart from its conserved F-box domain. Here we show that the SLF C-terminal region possesses a novel ubiquitin-binding domain （UBD） structure conserved among the SLF protein family. By using an ex vivo system of Nicotiana benthamiana, we found that the UBD mediates the SLF protein turnover by the ubiquitin-proteasome pathway. Furthermore, we detected that the SLF protein was directly involved in S-RNase degradation. Taken together, our results provide a novel insight into the SLF structure and highlight a potential role of SLF protein stability and degradation in S-RNase-based self-incompatibility.展开更多
The identification of self-incompatibility genotype （S-genotype） will be useful for selection of pollinizers and design of crossing in cultivar improvement of sand pear. This paper reported the identification of sel...The identification of self-incompatibility genotype （S-genotype） will be useful for selection of pollinizers and design of crossing in cultivar improvement of sand pear. This paper reported the identification of self-incompatibility genotypes of seven Chinese and two Japanese sand pear cultivars using PCR-RFLP analysis and S-RNase sequencing. The Sgenotypes of these cultivars were determined as follows： Huali 1 S1S3, Shounan S1S3, Xizilti S1S4, Qingxiang S3S7, Sanhua S2S7, Huangmi （Imamuranatsu） S1S6, Huali 2 S3S4, Baozhuli S7S33, Cangxixueli S5S15. S-RNase alleles （S1 to S9） in sand pear could be identified effectively by PCR-RFLP analysis.展开更多
66 F 1 hybrids, produced by 3 double low self-incompatible lines and 22 varieties with a North Carolina II (NCII) crossing design, were tested for their heterosis in Wuhan, China during two growing seasons from 1999 -...66 F 1 hybrids, produced by 3 double low self-incompatible lines and 22 varieties with a North Carolina II (NCII) crossing design, were tested for their heterosis in Wuhan, China during two growing seasons from 1999 - 2001. The results showed that significant differences were found between F1s and their parents for yield per plant and seed oil content. Mid-parent heterosis of these two characters ranged from 5.50% -64.11% and from 1.55% -7.44% respectively. Heterosis for seed yield per plant was greater than that of seed oil content. For yield components, heterosis of total number of siliques per plant was the highest, followed by seed number per silique and 1 000 seeds weight. Significant genotype-by-year interaction was found for seed yield per plant. Results from correlation and combining ability analysis indicated that parental effects on its F! hybrid depended on characters, seed yield per plant was affected by both additive and non-additive effects, and seed oil content was affected mainly by additive effect. When designing hybrid programme, parents might be selected by GCAs and variances of SCAs of parents for the characters affected by both additive and non-additive effects, and by the sum of GCAs of female and male parents for the characters mainly affected by additive effects.展开更多
[ Objective] This study aimed to explore the relationship between self-incompatibility strength and characteristics related to pollination and fertilization of different apricot varieties in Xinjiang. [ Method] The po...[ Objective] This study aimed to explore the relationship between self-incompatibility strength and characteristics related to pollination and fertilization of different apricot varieties in Xinjiang. [ Method] The pollen amount, pollen germination rate, pollen tube growth status and fruiting setting rate by self-pollina- tion of 34 apricot cultivars in Xinjiang were determined, to analyze the self-incompatibility of different apricot cultivars. [ Result] The average pollen amount per anther of 34 apricot eultivars was 1 213.7, and the average pollen germination rate was 46.0%. There were great differences in the self-incompatlbility of different cuhivars ; most pollen tubes of the euhivars with high self-incompatibility stopped elongating at 1/3 or 1/2 part of the styles, and only a few pollen tubes of the euhivars with low self-incompatibility reached the ovary, and the normal fertilization ratio was significantly lower than that in self-compatible cultivars. [ Conclusion] Among the 34 apricot cuhivars, only 6 cuhivars were self-compatible and the others exhibited gametophyte self-incompatibility. In addition, the fruit setting rate by self-pollination was low.展开更多
Plants have evolved diverse self-incompatibility(SI)systems for outcrossing.Since Darwin’s time,consid-erable progress has been made toward elucidating this unrivaled reproductive innovation.Recent advances in interd...Plants have evolved diverse self-incompatibility(SI)systems for outcrossing.Since Darwin’s time,consid-erable progress has been made toward elucidating this unrivaled reproductive innovation.Recent advances in interdisciplinary studies and applications of biotechnology have given rise to major break-throughs in understanding the molecular pathways that lead to SI,particularly the strikingly different SI mechanisms that operate in Solanaceae,Papaveraceae,Brassicaceae,and Primulaceae.These best-un-derstood SI systems,together with discoveries in other"nonmodel"SI taxa such as Poaceae,suggest a complex evolutionary trajectory of SI,with multiple independent origins and frequent and irreversible losses.Extensive exploration of self-/nonself-discrimination signaling cascades has revealed a compre-hensive catalog of male and female identity genes and modifier factors that control SI.Thesefindings also enable the characterization,validation,and manipulation of SI-related factors for crop improvement,helping to address the challenges associated with development of inbred lines.Here,we review current knowledge about the evolution of SI systems,summarize key achievements in the molecular basis of pol-len‒pistil interactions,discuss potential prospects for breeding of SI crops,and raise several unresolved questions that require further investigation.展开更多
Self-incompatibility(SI)is an intraspecific reproductive barrier widely present in angiosperms.The SI system with the broadest occurrence in angiosperms is based on an S-RNase linked to a cluster of multiple S-locus F...Self-incompatibility(SI)is an intraspecific reproductive barrier widely present in angiosperms.The SI system with the broadest occurrence in angiosperms is based on an S-RNase linked to a cluster of multiple S-locus F-box(SLF)genes found in the Solanaceae,Plantaginaceae,Rosaceae,and Rutaceae.Recent studies reveal that non-self S-RNase is degraded by the Skip Cullin F-box(SCF)SLF-mediated ubiquitin–proteasome system in a collaborative manner in Petunia,but how self-RNase functions largely remains mysterious.Here,we show that S-RNases form S-RNase condensates(SRCs)in the self-pollen tube cytoplasm through phase separation and the disruption of SRC formation breaks SI in self-incompatible Petunia hybrida.We further find that the pistil SI factors of a small asparagine-rich protein HT-B and thioredoxin h together with a reduced state of the pollen tube all promote the expansion of SRCs,which then sequester several actin-binding proteins,including the actin polymerization factor PhABRACL,the actin polymerization activity of which is reduced by S-RNase in vitro.Meanwhile,we find that S-RNase variants lacking condensation ability fail to recruit PhABRACL and are unable to induce actin foci formation required for pollen tube growth inhibition.Taken together,our results demonstrate that phase separation of S-RNase promotes SI response in P.hybrida,revealing a new mode of S-RNase action.展开更多
Gametophytic self-incompatibility （GSI） is controlled by a highly polymorphic locus called the S-locus, which is an important factor that can result in seedless fruit in Citrus. The S 1 self-incompatibility locus-li...Gametophytic self-incompatibility （GSI） is controlled by a highly polymorphic locus called the S-locus, which is an important factor that can result in seedless fruit in Citrus. The S 1 self-incompatibility locus-linked pollen 3.15 gene （S1-3.15） belongs to a type of S locus gene. The role of S1-3.15 in the SI reaction of Citrus has not yet been reported. In this study, full-length sequences of cDNA and DNA encoding the S1-3.15 gene, referred to as CrS1-3.15 , were isolated from ‘Wuzishatangju’ （Self-incompatibility, SI） and ‘Shatangju’ （Self-compatibility, SC） . The predicted amino acid sequences of CrS1-3.15 between ‘Wuzishatangju’ and ‘Shatangju’ differ by only three amino acids. Compared to ‘Wuzishatangju’, three bases were substituted in the genomic DNA of CrS1-3.15 from ‘Shatangju’. Southern blot results showed that one copy of CrS1-3.15 existed in the genomic DNA of both ‘Wuzishatangju’ and ‘Shatangju’. The expression level of the CrS1-3.15 gene in the ovaries of ‘Shatangju’ was approximately 60-fold higher than that in the ovaries of ‘Wuzishatangju’. When ‘Wuzishatangju’ was cross-pollinated, the expression of CrS1-3.15 was upregulated in the ovaries at 3d, and the highest expression levels were detected in the ovaries at 6d after cross-pollination of ‘Wuzishatangju’ × ‘Shatangju’. To obtain the CrS1-3.15 protein, the full-length cDNA of CrS1-3.15 genes from ‘Wuzishatangju’ and ‘Shatangju’ was successfully expressed in Pichia pastoris. Pollen germination frequency of ‘Wuzishatangju’ was inhibited significantly with increasing CrS1-3.15 protein concentrations from SI ‘Wuzishatangju’.展开更多
Common buckwheat(Fagopyrum esculentum)and Tartary buckwheat(Fagopyrum tataricum),the two most widely cultivated buckwheat species,differ greatly in flavonoid content and reproductive mode.Here,we report the first high...Common buckwheat(Fagopyrum esculentum)and Tartary buckwheat(Fagopyrum tataricum),the two most widely cultivated buckwheat species,differ greatly in flavonoid content and reproductive mode.Here,we report the first high-quality and chromosome-level genome assembly of common buckwheat with 1.2 Gb.Comparative genomic analysis revealed that common buckwheat underwent a burst of long terminal repeat retrotransposons insertion accompanied by numerous large chromosome rearrangements after divergence from Tartary buckwheat.Moreover,multiple gene families involved in stress tolerance and flavonoid biosynthesis such as multidrug and toxic compound extrusion(MATE)and chalcone synthase(CHS)underwent significant expansion in buckwheat,especially in common buckwheat.Integrated multi-omics analysis identified high expression of catechin biosynthesis-related genes in flower and seed in common buckwheat and high expression of rutin biosynthesis-related genes in seed in Tartary buckwheat as being important for the differences in flavonoid type and content between these buckwheat species.We also identified a candidate key rutindegrading enzyme gene(Ft8.2377)that was highly expressed in Tartary buckwheat seed.In addition,we identified a haplotype-resolved candidate locus containing many genes reportedly associated with the development of flower and pollen,which was potentially related to self-incompatibility in common buckwheat.Our study provides important resources facilitating future functional genomics-related research of flavonoid biosynthesis and selfincompatibility in buckwheat.展开更多
Pear has an S-RNase-based gametophytic selfincompatibility(SI)system.Nuclear DNA degradation is a typical feature of incompatible pollen tube death,and is among the many physiological functions of vacuoles.However,the...Pear has an S-RNase-based gametophytic selfincompatibility(SI)system.Nuclear DNA degradation is a typical feature of incompatible pollen tube death,and is among the many physiological functions of vacuoles.However,the specific changes that occur in vacuoles,as well as the associated regulatory mechanism in pear SI,are currently unclear.Although research in tobacco has shown that decreased activity of diacylglycerol kinase(DGK)results in the morphological change of pollen tube vacuole,whether DGK regulates the pollen tube vacuole of tree plants and whether it occurs in SI response,is currently unclear.We found that DGK activity is essential for pear pollen tube growth,and DGK4 regulates pollen tube vacuole morphology following its high expression and deposition at the tip and shank edge of the pollen tube of pear.Specifically,incompatible S-RNase may induce cytoplasmic acidification of the pollen tube by inhibiting V-ATPase V0 domain a1 subunit gene expression as early as 30 min after treatment,when the pollen tube is still alive.Cytoplasmic acidification induced by incompatible S-RNase results in reduced DGK4 abundance and deposition,leading to morphological change of the vacuole and fragmentation of nuclear DNA,which indicates that DGK4 is a key factor in pear SI response.展开更多
Cellular responses rely on signal perception and integration. A nice example of this is self incompatibility （SI）, which is an important mechanism to prevent inbreeding. It prevents self-fertilization by using a hig...Cellular responses rely on signal perception and integration. A nice example of this is self incompatibility （SI）, which is an important mechanism to prevent inbreeding. It prevents self-fertilization by using a highly discriminatory cellular recognition and rejection mechanism. Most Sl systems are genetically specified by the S-locus, which has a pollen and a pistil S-component. A receptor-ligand interaction is used by Papaver rhoeas to control SI. S proteins encoded by the pistil part of the S-locus interact with incompatible pollen to achieve rapid inhibition of tip growth. The incompatible Sl interaction triggers a Ca^2＋-dependent signaling cascade. A number of Sl-specific events are triggered in incompatible pollen, including rapid depolymerization of the actin cytoskeleton; phosphorylation of soluble inorganic pyrophosphatases （SPPases）, Prp26.1; activation of a mitogen activated protein kinase, p56; programmed cell death （PCD） involving a caspase-3-1ike activity. These events contribute to prevent self-fertilizaUon. We are attempting to establish the functional significance of these events, and their possible involvement in integrating a coordinated signaling response. Here we describe the identification of these components shown to be involved in Sl, together with recent progress in identifying links between some of them. These data constitute the first steps in elucidating how SI signaling is integrated.展开更多
Based on the c DNA sequences from hyper variable(HV) regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase c DNA probes were designed, and a c DNA microarray for S-RNase detections was established. Ea...Based on the c DNA sequences from hyper variable(HV) regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase c DNA probes were designed, and a c DNA microarray for S-RNase detections was established. Each microarray contained 240 sites from 55 c DNA probes, including all specific c DNA sequences from the HV regions of the S-alleles. Using the c DNA of pistils of tested pear cultivars as template and Cy3 fluorescently labeling primers by PCR amplification, microarray hybridization detected the S-genotype of each pear cultivar. The genotypes inferred from the c DNA microarray hybridization signals of pear cultivars such as ‘Lijiang Huangsuanli', ‘Xiuyu', ‘Midu Yuli', ‘Baimianli', and ‘Deshengxiang' were similar to the known genotypes of all tested cultivars. The S-RNase c DNA microarrays and the oligonucleotide gene chips were then used to conduct parallel testing of 24 P. pyrifolia cultivars with unknown S-genotypes. In conclusion, the construction of c DNA microarrays has further improved the pear S-RNase detection platform.展开更多
A new study provides a comprehensive molecular mechanism that controls interspecific incompatibility of self-incompatible(SI)plants in the Brassicaceae.This finding points to a potentially promising path to break inte...A new study provides a comprehensive molecular mechanism that controls interspecific incompatibility of self-incompatible(SI)plants in the Brassicaceae.This finding points to a potentially promising path to break interspecific barriers and achieve introgression of desirable traits into crops from distant species among SI crops in the Brassicaceae.展开更多
Self-incompatibility(SI)is an important genetic mechanism in angiosperms that prevents inbreeding and promotes outcrossing,with significant implications for crop breeding,including genetic diversity,hybrid seed produc...Self-incompatibility(SI)is an important genetic mechanism in angiosperms that prevents inbreeding and promotes outcrossing,with significant implications for crop breeding,including genetic diversity,hybrid seed production,and yield optimization.In eudicots,SI is typically governed by a single Slocus containing tightly linked pistil and pollen S-determinant genes.Despite major advances in SI research,a centralized,comprehensive resource for SI-related genomic data remains lacking.To address this gap,we developed the Plant Self-Incompatibility Atlas(PSIA),a systematically curated knowledgebase providing an extensive compilation of plant SI,including genomic resources for SI species,S gene annotations,molecular mechanisms,phylogenetic relationships,and comparative genomic analyses.The current release of PSIA includes over 500 genome assemblies from 469 SI species.Using known S genes as queries,we manually identified and rigorously curated 3700 S genes.PSIA provides detailed S-locus information from assembled genomes of SI species and offers an interactive platform for browsing,BLAST searches,S gene analysis,and data retrieval.Additionally,PSIA serves as a unique platform for comparative genomic studies of S-loci,facilitating exploration of the dynamic processes underlying the origin,loss,and regain of SI.As a comprehensive and user-friendly resource,PSIA will greatly advance our understanding of angiosperm SI and serve as a valuable tool for crop breeding and hybrid seed production.PSIA is freely available at http://www.plantsi.cn.展开更多
Identification of the S genotype of Malus plants will greatly promote the discovery of new genes,the cultivation and production of apple,the breeding of new varieties,and the origin and evolution of self-incompatibili...Identification of the S genotype of Malus plants will greatly promote the discovery of new genes,the cultivation and production of apple,the breeding of new varieties,and the origin and evolution of self-incompatibility in Malus plants.In this experiment,88 Malus germplasm resources,such as Aihuahong,Xishuhaitang,and Reguanzi,were used as materials.Seven gene-specific primer combinations were used in the genotype identification.PCR amplification using leaf DNA produced a single S-RNase gene fragment in all materials.The results revealed that 70 of the identified materials obtained a complete S-RNase genotype,while only one S-RNase gene was found in 18 of them.Through homology comparison and analysis,13 S-RNase genotypes were obtained:S_(1)S_(2)(Aihuahong,etc.),S_(1)S_(28)(Xixian Haitang,etc.),S_(1)S_(51)(Hebei Pingdinghaitang),S_(1)S_(3)(Xiangyangcun Daguo,etc.),S_(2)S_(3)(Zhaiyehaitang,etc.),S_(3)S_(51)(Xishan 1),S_(3)S_(28)(Huangselihaerde,etc.),S_(2)S_(28)(Honghaitang,etc.),S_(4)S_(28)(Bo 11),S_(7)S_(28)(Jiuquan Shaguo),S_(10)S_e(Dongchengguan 13),S_(10)S_(21)(Dongxiangjiao)and S_(3)S_(51)(Xiongyue Haitang).Simultaneously,the frequency of the S gene in the tested materials was analyzed.The findings revealed that different S genes had varying frequencies in Malus resources,as well as varying frequencies between intraspecific and interspecific.S_(3) had the highest frequency of 68.18%,followed by S_(1)(42.04%).In addition,the phylogenetic tree and origin evolution analysis revealed that the S gene differentiation was completed prior to the formation of various apple species,that cultivated species also evolved new S genes,and that the S_(50) gene is the oldest S allele in Malus plants.The S_(1),S_(29),and S_(33) genes in apple-cultivated species,on the other hand,may have originated in M.sieversii,M.hupehensis,and M.kansuensis,respectively.In addition to M.sieversii,M.kansuensis and M.sikkimensis may have also played a role in the origin and evolution of some Chinese apples.展开更多
Fertilization in flowering plants is completed through several recognitionevents, and the first of which is the recognition of pollen by pistil of female reproductivetissue. Self-incompatibility (SI) is an intraspecif...Fertilization in flowering plants is completed through several recognitionevents, and the first of which is the recognition of pollen by pistil of female reproductivetissue. Self-incompatibility (SI) is an intraspecific reproductive barrier to prevent selfferitilization and widely distributed in flowering plants. In many species, SI shows simplegenetics and is controlled by a single multi-allelic locus, called the S locus. In gametophyticSI (GSI) exemplified by the Solanaceae, Scrophulariaceae and Rosaceae, a class ofribonucleases, called S RNases, have been shown to mediate the stylar expression of SI butnot the pollen expression of SI. The latter appears to be determined by a gene differentfrom those encoding S RNases, often referred to as pollen S gene. The pollen S gene is thecrucial missing part in understanding the biochemical and molecular mechanisms of self andnon-self pollen recognition in flowering plants. Recent genetic analysis of mutationsaffecting the pollen expression of SI has suggested a possible model of how the pollen S geneinteracts with S RNases to achieve self and non-self pollen recognition. Furthermore, wewill present two approaches, S-locus directed transposon tagging and map-based cloning, forcloning the pollen S in Antirrhinum.展开更多
基金supported by grants from the National Natural Science Foundation of China(Grant No.31960585)Science and Technology Major Project of Guangxi(Grant No.Guike AA22068092)+1 种基金Guangxi Science and Technology Vanguard Special Action Project(Grant No.202204)State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources(Grant Nos.SKLCUSA-a201906,SKLCU-SA-c201901)。
文摘The SKP1 gene is an important component of the SCF(SKP1-Cullin1-F-box)complex and serves as a bridge connecting the F-box and Cullin1genes(F-box-SKP1-Cullin1).The pattern of S-RNase being ubiquitously labelled by the SCF complex and degraded by the 26S protease accounts for the bulk of the available self-incompatibility studies.In this study,15 ClSKP1s from the‘Xiangshui'lemon genome and ubiquitome exist in the same SKP1 conserved domain(CD)as SKP1s in other species.The q PCR results showed that SKP1-6 and SKP1-14 have tissue expression patterns specific for expression in pollen.In addition,SKP1-6 and SKP1-14 in the stigma,style and ovary were significantly upregulated after self-pollination compared to those after cross-pollination.A subcellular location showed that SKP1-6 and SKP1-14 were located in the nucleus.In addition,yeast two-hybrid(Y2H)assays,bimolecular fluorescence complementation(BiFC)and luciferase complementation imaging(LCI)assays showed that SKP1-6 interacted with F-box1,F-box33,F-box34,F-box17,F-box19,Cullin1-2 and 26S proteasome subunit 4 homolog A(26S PS4HA).SKP1-14 interacted with F-box17,F-box19,F-box35,Cullin1-2 and 26S PS4HA.The interaction of Cullin1-2 and the F-box with SKP1 as a bridge was verified by a yeast three-hybrid experiment.The ability of S3-RNase to inhibit pollen and pollen tube growth and development was assessed using in vitro pollen co-culture experiments with recombinant S3-RNase proteins.Overall,this study provides important experimental evidence and theoretical basis for understanding the mechanism of self-incompatibility in plants by revealing the key role of the SCF complex in‘Xiangshui'lemon,which is bridged by ClSKP1-6,in self-incompatibility.The results of this study are of great significance for the future indepth exploration of the molecular mechanism of the SCF complex and its wide application in the self-incompatibility of plants.
基金supported by the National Natural Science Foundation of China(32372695 and 32488302)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences(CAASZDRW202404)。
文摘Re-domestication of diploid potato(Solanum tuberosum)into a seed crop is an innovative breeding method to accelerate genetic improvement.Seed propagation would allow hybrid production and mix superior alleles.However,almost all diploid potatoes in nature are self-incompatible(SI).Gametophytic self-incompatible(GSI)is a widespread SI in Solanaceae and is controlled by the S locus that contains a ribonuclease(S-RNase)and multiple F-box(SLFs);however,the genetic diversity of the S locus in potato is unclear.This study identified 21 S-RNase alleles involved in SI from 194 diploid potato accessions by large-scale transcriptome sequencing.The levels of amino acid similarity among different S-RNase proteins varied from 31.3 to 95.8%.S2 allele is the most widespread in 194 diploid potatoes and is mainly distributed in the S.tuberosum Group Phureja.Based on genomic annotation and expression analysis,we identified 12 potential functional SI male-determinant genes,S-locus F-box(SLFs),encoding F-box proteins in the S2 locus on a genomic region of approximately 13 Mb.Comparative genomics analysis showed that eight SLF genes are relatively conserved among four homozygous S locus.The Ka and Ks analysis suggested that S-RNase and intra-haplotypic SLF genes have co-evolved.These findings help select suitable pollinators,combine more hybrid combinations,and fully use heterosis to accelerate diploid potato breeding.
基金Scientific Research Foundation of Science&Technology Department of Sichuan Province(2021YJ0497).
文摘In plants,heteromorphic self-incompatibility(HetSI)is a strategy for avoiding self-pollination and promoting outcrossing,and during this process,numerous protein-protein interaction events occur between the pistil and pollen.Previous studies in Primula and Fagopyrum that focused on HetSI systems have provided interesting insights;however,the molecular mechanism underlying HetSI remains largely unknown.In this study,we profiled the proteome of Plumbago auriculata stigmas before and after self-incompatible(SI)and self-compatible(SC)pollination.Comparative analyses were conducted by 4D-DIA(Four-dimensional data independent acquisition),a promising technology that increases the sensitivity and reduces the spectral complexity of proteomic analysis by adding a fourth dimension,ion mobility.The results revealed 33387 peptides and 5311 proteins in all samples.The pathways in which the differentially expressed proteins(DEPs)identified in the P×P(Pin style self-pollinated with pin pollen)vs.PS(Pin style)and T×T(Thrum style self-pollinated with thrum pollen)vs.TS(Thrum style)comparisons were significantly enriched were biosynthesis of secondary metabolites and pentose and glucuronate interconversions.In the P×T(Pin style cross-pollinated with thrum pollen)vs.PS and T×P(Thrum style cross-pollinated with pin pollen)vs.TS comparison,the top three pathways were biosynthesis of secondary metabolites,pentose and glucuronate interconversions,and phenylpropanoid biosynthesis.The phenylpropanoid biosynthesis,cutin,suberine and wax biosynthesis,and flavonoid biosynthesis pathways were enriched in the P×T vs.P×P comparison,and starch and sucrose metabolism,glycerophospholipid metabolism,and alpha-linolenic acid metabolism were abundant in the T×T vs.T×P comparison.The enriched pathways between PS and TS were the biosynthesis of secondary metabolites,phenylpropanoid biosynthesis,and pentose and glucuronate interconversion.Self-incompatibility protein S1(SI S1),Mitogen-activated protein kinase 3/4(MPK3/4),Mitogen-activated protein kinase kinase 2/3(M2K2/3),Exocyst complex component EXO70A1(E70A1)and Thioredoxin H1/2(TRXH1/2)were found to be HetSI-related candidates,and O-fucosyltransferase 23(OFT23),3-ketoacyl-CoA synthase 6(KCS6),Receptor-like protein kinase FERONIA(FERON),Fimbrin-5(FIMB5),Pollen-specific leucine-rich repeat extensin-like protein 4(PLRX4),Transcription initiation factor IIB-2(TF2B2)and Pectinesterase 1(AL11A),etc.,were identified as other regulatory transducers.These findings combined with our morphological and reactive oxygen species(ROS)intensity analyses indicate that P.auriculata has typical dry-stigmas and that the HetSI mechanism might differ between the pin and thrum.SI S1 might be the key factor in HetSI,and ROS are overexpressed during SC pollination to rapidly activate the mitogen-activated protein kinase(MAPK)-mediated phosphorylation of E70A1 to maintain stigma receptivity in plants with HetSI.
文摘The self-incompatibility ( S) loci from the Solanaceae, Rosaceae and Scrophulariaceae encode a class of ribonucleases, known as S RNases, which have been shown to control the pistil expression of self-incompatible reaction. In the former two families, the S loci have been shown to be located near centromere. However, the chromosomal location of the S locus in Antirrhinum, a species of the Scrophulariaceae, is not known. To determine its chromosomal location and genomic organization, an S-2 RNase gene and its corresponding 63 kb BAC clone were separately used for fluorescence in situ hybridization (FISH) of mitotic metaphase chromosomes of a self-incompatible Antirrhinum line Of S2S5. The results showed that the S-2 RNase detected a doublet signal near the centromere of the smallest chromosome (2n = 16). Two separate doublet signals of the tested BAC sequence were shown on both sides of the centromeres of all eight pairs of the chromosomes, suggesting that the Antirrhinum S locus is located in a pericentromeric region. Furthermore, a retrotransposon, named RIS1 (retrotransposon in the S locus), which has not been identified yet in. Antirrhinum, was found next to S-2 RNase. Taken together, the centromeric location of the S locus from the three S-RNase-based self-incompatible families provides a further support on a common origin of their evolution as well as suppressed recombination.
基金supported by the National Basic Research Program of China(973 Program)(Nos.2007CB108703 and 2011CB915404)the National Natural Science Foundation of China(No.30921003)
文摘In flowering plants, self-incompatibility （SI） serves as an important intraspecific reproductive barrier to promote outbreeding. In species from the Solanaceae, Plantaginaceae and Rosaceae, S-RNase and SLF （S-locus F-box） proteins have been shown to control the female and male specificity of SI, respectively. However, little is known about structure features of the SLF protein apart from its conserved F-box domain. Here we show that the SLF C-terminal region possesses a novel ubiquitin-binding domain （UBD） structure conserved among the SLF protein family. By using an ex vivo system of Nicotiana benthamiana, we found that the UBD mediates the SLF protein turnover by the ubiquitin-proteasome pathway. Furthermore, we detected that the SLF protein was directly involved in S-RNase degradation. Taken together, our results provide a novel insight into the SLF structure and highlight a potential role of SLF protein stability and degradation in S-RNase-based self-incompatibility.
基金supported in part by Natural Science Foundation of JiangxiAgricultural University, China (1878).
文摘The identification of self-incompatibility genotype （S-genotype） will be useful for selection of pollinizers and design of crossing in cultivar improvement of sand pear. This paper reported the identification of self-incompatibility genotypes of seven Chinese and two Japanese sand pear cultivars using PCR-RFLP analysis and S-RNase sequencing. The Sgenotypes of these cultivars were determined as follows： Huali 1 S1S3, Shounan S1S3, Xizilti S1S4, Qingxiang S3S7, Sanhua S2S7, Huangmi （Imamuranatsu） S1S6, Huali 2 S3S4, Baozhuli S7S33, Cangxixueli S5S15. S-RNase alleles （S1 to S9） in sand pear could be identified effectively by PCR-RFLP analysis.
基金supported by the National Key Basic Research Special Foundation of China(2001CB10807)European Union Project(IC-18-CT97-0172)China-Sweden Cooperation Project(Plan for breeding SI hybrids).
文摘66 F 1 hybrids, produced by 3 double low self-incompatible lines and 22 varieties with a North Carolina II (NCII) crossing design, were tested for their heterosis in Wuhan, China during two growing seasons from 1999 - 2001. The results showed that significant differences were found between F1s and their parents for yield per plant and seed oil content. Mid-parent heterosis of these two characters ranged from 5.50% -64.11% and from 1.55% -7.44% respectively. Heterosis for seed yield per plant was greater than that of seed oil content. For yield components, heterosis of total number of siliques per plant was the highest, followed by seed number per silique and 1 000 seeds weight. Significant genotype-by-year interaction was found for seed yield per plant. Results from correlation and combining ability analysis indicated that parental effects on its F! hybrid depended on characters, seed yield per plant was affected by both additive and non-additive effects, and seed oil content was affected mainly by additive effect. When designing hybrid programme, parents might be selected by GCAs and variances of SCAs of parents for the characters affected by both additive and non-additive effects, and by the sum of GCAs of female and male parents for the characters mainly affected by additive effects.
基金Supported by Science and Technology Project of Xinjiang Uygur Autonomous Region"ResearchDemonstration of Utilization Technology of Special Fruit Tree Germplasm Resources in Xinjiang"(201130102-1)Key Discipline Pomology in Xinjiang Uygur Autonomous Region
文摘[ Objective] This study aimed to explore the relationship between self-incompatibility strength and characteristics related to pollination and fertilization of different apricot varieties in Xinjiang. [ Method] The pollen amount, pollen germination rate, pollen tube growth status and fruiting setting rate by self-pollina- tion of 34 apricot cultivars in Xinjiang were determined, to analyze the self-incompatibility of different apricot cultivars. [ Result] The average pollen amount per anther of 34 apricot eultivars was 1 213.7, and the average pollen germination rate was 46.0%. There were great differences in the self-incompatlbility of different cuhivars ; most pollen tubes of the euhivars with high self-incompatibility stopped elongating at 1/3 or 1/2 part of the styles, and only a few pollen tubes of the euhivars with low self-incompatibility reached the ovary, and the normal fertilization ratio was significantly lower than that in self-compatible cultivars. [ Conclusion] Among the 34 apricot cuhivars, only 6 cuhivars were self-compatible and the others exhibited gametophyte self-incompatibility. In addition, the fruit setting rate by self-pollination was low.
基金supported by the Forestry Peak Discipline Construction Project of Fujian Agriculture and Forestry University (Grant number 72202200205)。
文摘Plants have evolved diverse self-incompatibility(SI)systems for outcrossing.Since Darwin’s time,consid-erable progress has been made toward elucidating this unrivaled reproductive innovation.Recent advances in interdisciplinary studies and applications of biotechnology have given rise to major break-throughs in understanding the molecular pathways that lead to SI,particularly the strikingly different SI mechanisms that operate in Solanaceae,Papaveraceae,Brassicaceae,and Primulaceae.These best-un-derstood SI systems,together with discoveries in other"nonmodel"SI taxa such as Poaceae,suggest a complex evolutionary trajectory of SI,with multiple independent origins and frequent and irreversible losses.Extensive exploration of self-/nonself-discrimination signaling cascades has revealed a compre-hensive catalog of male and female identity genes and modifier factors that control SI.Thesefindings also enable the characterization,validation,and manipulation of SI-related factors for crop improvement,helping to address the challenges associated with development of inbred lines.Here,we review current knowledge about the evolution of SI systems,summarize key achievements in the molecular basis of pol-len‒pistil interactions,discuss potential prospects for breeding of SI crops,and raise several unresolved questions that require further investigation.
基金supported by the National Natural Science Foundation of China(32030007)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB27010302).
文摘Self-incompatibility(SI)is an intraspecific reproductive barrier widely present in angiosperms.The SI system with the broadest occurrence in angiosperms is based on an S-RNase linked to a cluster of multiple S-locus F-box(SLF)genes found in the Solanaceae,Plantaginaceae,Rosaceae,and Rutaceae.Recent studies reveal that non-self S-RNase is degraded by the Skip Cullin F-box(SCF)SLF-mediated ubiquitin–proteasome system in a collaborative manner in Petunia,but how self-RNase functions largely remains mysterious.Here,we show that S-RNases form S-RNase condensates(SRCs)in the self-pollen tube cytoplasm through phase separation and the disruption of SRC formation breaks SI in self-incompatible Petunia hybrida.We further find that the pistil SI factors of a small asparagine-rich protein HT-B and thioredoxin h together with a reduced state of the pollen tube all promote the expansion of SRCs,which then sequester several actin-binding proteins,including the actin polymerization factor PhABRACL,the actin polymerization activity of which is reduced by S-RNase in vitro.Meanwhile,we find that S-RNase variants lacking condensation ability fail to recruit PhABRACL and are unable to induce actin foci formation required for pollen tube growth inhibition.Taken together,our results demonstrate that phase separation of S-RNase promotes SI response in P.hybrida,revealing a new mode of S-RNase action.
基金supported by the National Natural Science Foundation of China(31000899)the Research Fund for the Doctoral Program of Higher Education of China(20104404120015 and 20114404110018)+4 种基金the Guangdong Province Science Foundation of China(06025843)the Science and Technology Planning Project of Guangzhou(2010r1-C771)the open foundation of the State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,South China Agricultural University(KSL-CUSAb-2012-09)the Key Laboratory of Innovation and Utilization for Germplasm Resources in Horticultural Crops in Southern China of Guangdong Higher Education Institutes,South China Agricultural University(No.KBL11008)the "211" Construction Fund for Key Subjects of College of Horticulture,South China Agricultural University
文摘Gametophytic self-incompatibility （GSI） is controlled by a highly polymorphic locus called the S-locus, which is an important factor that can result in seedless fruit in Citrus. The S 1 self-incompatibility locus-linked pollen 3.15 gene （S1-3.15） belongs to a type of S locus gene. The role of S1-3.15 in the SI reaction of Citrus has not yet been reported. In this study, full-length sequences of cDNA and DNA encoding the S1-3.15 gene, referred to as CrS1-3.15 , were isolated from ‘Wuzishatangju’ （Self-incompatibility, SI） and ‘Shatangju’ （Self-compatibility, SC） . The predicted amino acid sequences of CrS1-3.15 between ‘Wuzishatangju’ and ‘Shatangju’ differ by only three amino acids. Compared to ‘Wuzishatangju’, three bases were substituted in the genomic DNA of CrS1-3.15 from ‘Shatangju’. Southern blot results showed that one copy of CrS1-3.15 existed in the genomic DNA of both ‘Wuzishatangju’ and ‘Shatangju’. The expression level of the CrS1-3.15 gene in the ovaries of ‘Shatangju’ was approximately 60-fold higher than that in the ovaries of ‘Wuzishatangju’. When ‘Wuzishatangju’ was cross-pollinated, the expression of CrS1-3.15 was upregulated in the ovaries at 3d, and the highest expression levels were detected in the ovaries at 6d after cross-pollination of ‘Wuzishatangju’ × ‘Shatangju’. To obtain the CrS1-3.15 protein, the full-length cDNA of CrS1-3.15 genes from ‘Wuzishatangju’ and ‘Shatangju’ was successfully expressed in Pichia pastoris. Pollen germination frequency of ‘Wuzishatangju’ was inhibited significantly with increasing CrS1-3.15 protein concentrations from SI ‘Wuzishatangju’.
基金supported by the National Natural Science Foundation of China(32100500)Natural Science Foundation of Hebei Province(C2021201048)supported by the fund from the State Key Laboratory of Plant Genomics and the Chinese Academy of Sciences(CASWX2021SF-0109)。
文摘Common buckwheat(Fagopyrum esculentum)and Tartary buckwheat(Fagopyrum tataricum),the two most widely cultivated buckwheat species,differ greatly in flavonoid content and reproductive mode.Here,we report the first high-quality and chromosome-level genome assembly of common buckwheat with 1.2 Gb.Comparative genomic analysis revealed that common buckwheat underwent a burst of long terminal repeat retrotransposons insertion accompanied by numerous large chromosome rearrangements after divergence from Tartary buckwheat.Moreover,multiple gene families involved in stress tolerance and flavonoid biosynthesis such as multidrug and toxic compound extrusion(MATE)and chalcone synthase(CHS)underwent significant expansion in buckwheat,especially in common buckwheat.Integrated multi-omics analysis identified high expression of catechin biosynthesis-related genes in flower and seed in common buckwheat and high expression of rutin biosynthesis-related genes in seed in Tartary buckwheat as being important for the differences in flavonoid type and content between these buckwheat species.We also identified a candidate key rutindegrading enzyme gene(Ft8.2377)that was highly expressed in Tartary buckwheat seed.In addition,we identified a haplotype-resolved candidate locus containing many genes reportedly associated with the development of flower and pollen,which was potentially related to self-incompatibility in common buckwheat.Our study provides important resources facilitating future functional genomics-related research of flavonoid biosynthesis and selfincompatibility in buckwheat.
基金This work was supported by the National Key Research and Development Program of China(2018YFD1000107)National Natural Science Foundation of China(32001983).
文摘Pear has an S-RNase-based gametophytic selfincompatibility(SI)system.Nuclear DNA degradation is a typical feature of incompatible pollen tube death,and is among the many physiological functions of vacuoles.However,the specific changes that occur in vacuoles,as well as the associated regulatory mechanism in pear SI,are currently unclear.Although research in tobacco has shown that decreased activity of diacylglycerol kinase(DGK)results in the morphological change of pollen tube vacuole,whether DGK regulates the pollen tube vacuole of tree plants and whether it occurs in SI response,is currently unclear.We found that DGK activity is essential for pear pollen tube growth,and DGK4 regulates pollen tube vacuole morphology following its high expression and deposition at the tip and shank edge of the pollen tube of pear.Specifically,incompatible S-RNase may induce cytoplasmic acidification of the pollen tube by inhibiting V-ATPase V0 domain a1 subunit gene expression as early as 30 min after treatment,when the pollen tube is still alive.Cytoplasmic acidification induced by incompatible S-RNase results in reduced DGK4 abundance and deposition,leading to morphological change of the vacuole and fragmentation of nuclear DNA,which indicates that DGK4 is a key factor in pear SI response.
基金Supported by the Biotechnology and Biological Sciences Research Council(B.B.S.R.C.).
文摘Cellular responses rely on signal perception and integration. A nice example of this is self incompatibility （SI）, which is an important mechanism to prevent inbreeding. It prevents self-fertilization by using a highly discriminatory cellular recognition and rejection mechanism. Most Sl systems are genetically specified by the S-locus, which has a pollen and a pistil S-component. A receptor-ligand interaction is used by Papaver rhoeas to control SI. S proteins encoded by the pistil part of the S-locus interact with incompatible pollen to achieve rapid inhibition of tip growth. The incompatible Sl interaction triggers a Ca^2＋-dependent signaling cascade. A number of Sl-specific events are triggered in incompatible pollen, including rapid depolymerization of the actin cytoskeleton; phosphorylation of soluble inorganic pyrophosphatases （SPPases）, Prp26.1; activation of a mitogen activated protein kinase, p56; programmed cell death （PCD） involving a caspase-3-1ike activity. These events contribute to prevent self-fertilizaUon. We are attempting to establish the functional significance of these events, and their possible involvement in integrating a coordinated signaling response. Here we describe the identification of these components shown to be involved in Sl, together with recent progress in identifying links between some of them. These data constitute the first steps in elucidating how SI signaling is integrated.
基金supported by the National Natural Science Foundation of China(31272124)
文摘Based on the c DNA sequences from hyper variable(HV) regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase c DNA probes were designed, and a c DNA microarray for S-RNase detections was established. Each microarray contained 240 sites from 55 c DNA probes, including all specific c DNA sequences from the HV regions of the S-alleles. Using the c DNA of pistils of tested pear cultivars as template and Cy3 fluorescently labeling primers by PCR amplification, microarray hybridization detected the S-genotype of each pear cultivar. The genotypes inferred from the c DNA microarray hybridization signals of pear cultivars such as ‘Lijiang Huangsuanli', ‘Xiuyu', ‘Midu Yuli', ‘Baimianli', and ‘Deshengxiang' were similar to the known genotypes of all tested cultivars. The S-RNase c DNA microarrays and the oligonucleotide gene chips were then used to conduct parallel testing of 24 P. pyrifolia cultivars with unknown S-genotypes. In conclusion, the construction of c DNA microarrays has further improved the pear S-RNase detection platform.
文摘A new study provides a comprehensive molecular mechanism that controls interspecific incompatibility of self-incompatible(SI)plants in the Brassicaceae.This finding points to a potentially promising path to break interspecific barriers and achieve introgression of desirable traits into crops from distant species among SI crops in the Brassicaceae.
基金supported by the National Natural Science Foundation of China(Grant Nos.32030007 and 32200273).
文摘Self-incompatibility(SI)is an important genetic mechanism in angiosperms that prevents inbreeding and promotes outcrossing,with significant implications for crop breeding,including genetic diversity,hybrid seed production,and yield optimization.In eudicots,SI is typically governed by a single Slocus containing tightly linked pistil and pollen S-determinant genes.Despite major advances in SI research,a centralized,comprehensive resource for SI-related genomic data remains lacking.To address this gap,we developed the Plant Self-Incompatibility Atlas(PSIA),a systematically curated knowledgebase providing an extensive compilation of plant SI,including genomic resources for SI species,S gene annotations,molecular mechanisms,phylogenetic relationships,and comparative genomic analyses.The current release of PSIA includes over 500 genome assemblies from 469 SI species.Using known S genes as queries,we manually identified and rigorously curated 3700 S genes.PSIA provides detailed S-locus information from assembled genomes of SI species and offers an interactive platform for browsing,BLAST searches,S gene analysis,and data retrieval.Additionally,PSIA serves as a unique platform for comparative genomic studies of S-loci,facilitating exploration of the dynamic processes underlying the origin,loss,and regain of SI.As a comprehensive and user-friendly resource,PSIA will greatly advance our understanding of angiosperm SI and serve as a valuable tool for crop breeding and hybrid seed production.PSIA is freely available at http://www.plantsi.cn.
基金financially supported by the Agricultural Science and Technology Innovation Program(CAASASTIP-2021-RIP-02)。
文摘Identification of the S genotype of Malus plants will greatly promote the discovery of new genes,the cultivation and production of apple,the breeding of new varieties,and the origin and evolution of self-incompatibility in Malus plants.In this experiment,88 Malus germplasm resources,such as Aihuahong,Xishuhaitang,and Reguanzi,were used as materials.Seven gene-specific primer combinations were used in the genotype identification.PCR amplification using leaf DNA produced a single S-RNase gene fragment in all materials.The results revealed that 70 of the identified materials obtained a complete S-RNase genotype,while only one S-RNase gene was found in 18 of them.Through homology comparison and analysis,13 S-RNase genotypes were obtained:S_(1)S_(2)(Aihuahong,etc.),S_(1)S_(28)(Xixian Haitang,etc.),S_(1)S_(51)(Hebei Pingdinghaitang),S_(1)S_(3)(Xiangyangcun Daguo,etc.),S_(2)S_(3)(Zhaiyehaitang,etc.),S_(3)S_(51)(Xishan 1),S_(3)S_(28)(Huangselihaerde,etc.),S_(2)S_(28)(Honghaitang,etc.),S_(4)S_(28)(Bo 11),S_(7)S_(28)(Jiuquan Shaguo),S_(10)S_e(Dongchengguan 13),S_(10)S_(21)(Dongxiangjiao)and S_(3)S_(51)(Xiongyue Haitang).Simultaneously,the frequency of the S gene in the tested materials was analyzed.The findings revealed that different S genes had varying frequencies in Malus resources,as well as varying frequencies between intraspecific and interspecific.S_(3) had the highest frequency of 68.18%,followed by S_(1)(42.04%).In addition,the phylogenetic tree and origin evolution analysis revealed that the S gene differentiation was completed prior to the formation of various apple species,that cultivated species also evolved new S genes,and that the S_(50) gene is the oldest S allele in Malus plants.The S_(1),S_(29),and S_(33) genes in apple-cultivated species,on the other hand,may have originated in M.sieversii,M.hupehensis,and M.kansuensis,respectively.In addition to M.sieversii,M.kansuensis and M.sikkimensis may have also played a role in the origin and evolution of some Chinese apples.
文摘Fertilization in flowering plants is completed through several recognitionevents, and the first of which is the recognition of pollen by pistil of female reproductivetissue. Self-incompatibility (SI) is an intraspecific reproductive barrier to prevent selfferitilization and widely distributed in flowering plants. In many species, SI shows simplegenetics and is controlled by a single multi-allelic locus, called the S locus. In gametophyticSI (GSI) exemplified by the Solanaceae, Scrophulariaceae and Rosaceae, a class ofribonucleases, called S RNases, have been shown to mediate the stylar expression of SI butnot the pollen expression of SI. The latter appears to be determined by a gene differentfrom those encoding S RNases, often referred to as pollen S gene. The pollen S gene is thecrucial missing part in understanding the biochemical and molecular mechanisms of self andnon-self pollen recognition in flowering plants. Recent genetic analysis of mutationsaffecting the pollen expression of SI has suggested a possible model of how the pollen S geneinteracts with S RNases to achieve self and non-self pollen recognition. Furthermore, wewill present two approaches, S-locus directed transposon tagging and map-based cloning, forcloning the pollen S in Antirrhinum.
