Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system has been widely used for precise gene editing in plants.However,simultaneous gene editing of multiple homoeoalleles r...Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system has been widely used for precise gene editing in plants.However,simultaneous gene editing of multiple homoeoalleles remains challenging,especially in self-incompatible polyploid plants.Here,we simultaneously introduced targeted mutations in all three homoeoalleles of two genes in the self-incompatible allohexaploid tall fescue,using both CRISPR/Cas9 and LbCas12a(LbCpf1)systems.Loss-of-function mutants of FaPDS exhibited albino leaves,while knockout of FaHSP17.9 resulted in impaired heat resistance in T0 generation of tall fescue.Moreover,these mutations were inheritable.Our findings demonstrate the feasibility of generating loss-of-function mutants in T0 generation polyploid perennial grasses using CRISPR/Cas systems.展开更多
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
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
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
[ 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
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
[ Objective] This study aimed to clone the PsSFBB gene from Xinjiang pear for bioinformatics analysis. [ Method ] PsSFBB gene was cloned from an- thers of Qipan pear by using RT-PCR and RACE technologies for bioinform...[ Objective] This study aimed to clone the PsSFBB gene from Xinjiang pear for bioinformatics analysis. [ Method ] PsSFBB gene was cloned from an- thers of Qipan pear by using RT-PCR and RACE technologies for bioinformatics analysis. [Result] A SFBBt-α gene with a full-length of 1 231 bp was cloned and named PsSFBB6-α （Genbank accession number： EU909685）. PsSFBBt-ct gene encodes a protein of 378 amino acids, with an F-box motif composed of about 50 amino acids in the N-temfinal. According to the bioinformatics analysis, the molecular formula of PsSFB6-α protein is C2000 H3034 N517 O558S223, with relative molecu- lar mass of 43 987.5 and isoelectric point of 6.02, and the secondary structure is dominated by or-helices ; theoretically, the half life period is 30 h and the instabil- ity parameter is 55.21, so PsSFBB6-α protein is an instable protein ; in addition, it is predicted that PsSFBB6-α protein is a hydrophilic and non-secreted protein with lyases activity and specifically recognized substrates, which was consistent with the function of F-box protein. [ Conclusion] This study laid the foundation for further research on SFBB proteins and the mechanism of self-incompatibility and provided theoretical basis for breeding of self-compatible cultivars of Xinjiang pear and scientific arrangement of pollination trees in production to increase the yield and quality.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
The exocyst is a conserved protein complex,and required for vesicles tethering,fusion and polarized exocytosis.Exo70A1,the exocyst subunit,is essential for assembly of the exocyst complex.To better understand potentia...The exocyst is a conserved protein complex,and required for vesicles tethering,fusion and polarized exocytosis.Exo70A1,the exocyst subunit,is essential for assembly of the exocyst complex.To better understand potential roles of Exo70A1 in non-heading Chinese cabbage（Brassica campestris ssp.chinensis）,we obtained the full-length cDNA of Exo70A1 gene,which consisted of 1 917 bp and encoded a protein of 638 amino acids.BlastX showed BcExo70A1 shared 94.9% identity with Brassica oleracea var.acephala（AEI26267.1）,and clustered into a same group with other homologues in B.oleracea var.acephala and Brassica napus.Subcellular localization analysis showed BcExo70A1 was localized to punctate structures in cytosol of onion epithelial cells.Results showed that BcExo70A1 was widely presented in stamens,young stems,petals,unpollinated pistils,roots and leaves of self compatible and incompatible plants.The transcripts of BcExo70A1 in non- heading Chinese cabbage declined during initial 1.5 h after incompatible pollination,while an opposite trend was presented after compatible pollination.Our study reveals that BcExo70A1 could play essential roles in plant growth and development,and is related to the rejection of self pollen in non-heading Chinese cabbage.展开更多
Pears carry a gametophytic self-incompatibility(SI)system.In this system,S-RNase is the SI pistil determinant,and S-locus F-box brothers(SFBBs)are candidate pollen determinants.However,compared with apple,fewer SFBB g...Pears carry a gametophytic self-incompatibility(SI)system.In this system,S-RNase is the SI pistil determinant,and S-locus F-box brothers(SFBBs)are candidate pollen determinants.However,compared with apple,fewer SFBB genes were identified from pear,possibly caused by the lack of economic and effective methods.Here,we used transcriptome sequencing on‘Yali’(Pyrus bretschneideri)to obtain sequence fragments of SFBB genes and then used polymerase chain reaction(PCR)to amplify the whole sequence of SFBB genes.Twenty-seven SFBB genes,including22 full-length and five nonfull-length SFBB genes,were identified in‘Yali’(P.bretschneideri).SFBBs linkage analysis by PCR-enzyme-linked immunoassay(ELISA)showed that 12 SFBB genes belong to the S21 locus,and 15 SFBB genes belong to the S34 locus.Phylogenetic analysis showed that SFBB genes from Pyrus were divided into 26 types,more than the original eight types.The intrahaplotypic divergence of SFBBs is high and comparable to the allelic diversity of S-RNase,which is consistent with a nonself-recognition SI system.In addition,the expression level of PbrSFBBs in‘Jinzhui’,the only known haploid pollen of a self-compatible mutant,was mostly approximately two times higher than in‘Yali’,which may be the reason for the self-compatible mutant.展开更多
Brassica rapa cuhivars Qinghaidahuang and Menyuan small rape were used as female parents and separately interspecifically hybridized with Zhongchijie- lan to artificially synthesize a number of Brasica napus resources...Brassica rapa cuhivars Qinghaidahuang and Menyuan small rape were used as female parents and separately interspecifically hybridized with Zhongchijie- lan to artificially synthesize a number of Brasica napus resources. Observation of meiosis, pollen fertility and pollen tube germination and analysis of self-compati- bility index of these artificial B. napus varieties were conducted. Results showed that pollen mother cells of artificial B. napus lines from two cross combinations all had normal chromosome behavior and high pollen fertility, but the selfing seed-setting and self-compatibility index varied extremely significantly; B. napus lines from cross combination Qinghaidahuang x Zhongchijidan are highly self-compatible, while B. napus lines from cross combination Menyuan small rape x Zhongchijielan are highly self-incompatible. This study analyzed the heredity laws of self-incompatibility trait in artificially synthesized lines and discussed applica- tion prospects of artificially synthesized self-incompatible B. napus lines in rapeseed production.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
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
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
基金This work was supported by the National Natural Science Foundation of China(31772349,31672482,and 31401915)the Major Science and Technology Innovation Project of Shandong Province(2019JZZY010726).
文摘Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system has been widely used for precise gene editing in plants.However,simultaneous gene editing of multiple homoeoalleles remains challenging,especially in self-incompatible polyploid plants.Here,we simultaneously introduced targeted mutations in all three homoeoalleles of two genes in the self-incompatible allohexaploid tall fescue,using both CRISPR/Cas9 and LbCas12a(LbCpf1)systems.Loss-of-function mutants of FaPDS exhibited albino leaves,while knockout of FaHSP17.9 resulted in impaired heat resistance in T0 generation of tall fescue.Moreover,these mutations were inheritable.Our findings demonstrate the feasibility of generating loss-of-function mutants in T0 generation polyploid perennial grasses using CRISPR/Cas systems.
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.
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.
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.
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.
基金Supported by National Natural Science Foundation of China(30860172)
文摘[ Objective] This study aimed to clone the PsSFBB gene from Xinjiang pear for bioinformatics analysis. [ Method ] PsSFBB gene was cloned from an- thers of Qipan pear by using RT-PCR and RACE technologies for bioinformatics analysis. [Result] A SFBBt-α gene with a full-length of 1 231 bp was cloned and named PsSFBB6-α （Genbank accession number： EU909685）. PsSFBBt-ct gene encodes a protein of 378 amino acids, with an F-box motif composed of about 50 amino acids in the N-temfinal. According to the bioinformatics analysis, the molecular formula of PsSFB6-α protein is C2000 H3034 N517 O558S223, with relative molecu- lar mass of 43 987.5 and isoelectric point of 6.02, and the secondary structure is dominated by or-helices ; theoretically, the half life period is 30 h and the instabil- ity parameter is 55.21, so PsSFBB6-α protein is an instable protein ; in addition, it is predicted that PsSFBB6-α protein is a hydrophilic and non-secreted protein with lyases activity and specifically recognized substrates, which was consistent with the function of F-box protein. [ Conclusion] This study laid the foundation for further research on SFBB proteins and the mechanism of self-incompatibility and provided theoretical basis for breeding of self-compatible cultivars of Xinjiang pear and scientific arrangement of pollination trees in production to increase the yield and quality.
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.
基金supported by the National Basic Research Program of China(973 Program,2009CB119001)the Fundamental Research Funds for the Central Universities of China(KYZ201111)+1 种基金the Natural Science Foundation of Jiangsu Province,China(BK2011643)the Scientific Innovation Research of College Graduate of Jiangsu Province,China(CXLX11_0688)
文摘The exocyst is a conserved protein complex,and required for vesicles tethering,fusion and polarized exocytosis.Exo70A1,the exocyst subunit,is essential for assembly of the exocyst complex.To better understand potential roles of Exo70A1 in non-heading Chinese cabbage（Brassica campestris ssp.chinensis）,we obtained the full-length cDNA of Exo70A1 gene,which consisted of 1 917 bp and encoded a protein of 638 amino acids.BlastX showed BcExo70A1 shared 94.9% identity with Brassica oleracea var.acephala（AEI26267.1）,and clustered into a same group with other homologues in B.oleracea var.acephala and Brassica napus.Subcellular localization analysis showed BcExo70A1 was localized to punctate structures in cytosol of onion epithelial cells.Results showed that BcExo70A1 was widely presented in stamens,young stems,petals,unpollinated pistils,roots and leaves of self compatible and incompatible plants.The transcripts of BcExo70A1 in non- heading Chinese cabbage declined during initial 1.5 h after incompatible pollination,while an opposite trend was presented after compatible pollination.Our study reveals that BcExo70A1 could play essential roles in plant growth and development,and is related to the rejection of self pollen in non-heading Chinese cabbage.
基金supported by the National Key Research and Development Program of China(Grant No.2018YFD1000107)National Natural Science Foundation of China(Grant No.32001983)the Natural Science Foundation of Jiangsu Province(Grant No.BK20190896)。
文摘Pears carry a gametophytic self-incompatibility(SI)system.In this system,S-RNase is the SI pistil determinant,and S-locus F-box brothers(SFBBs)are candidate pollen determinants.However,compared with apple,fewer SFBB genes were identified from pear,possibly caused by the lack of economic and effective methods.Here,we used transcriptome sequencing on‘Yali’(Pyrus bretschneideri)to obtain sequence fragments of SFBB genes and then used polymerase chain reaction(PCR)to amplify the whole sequence of SFBB genes.Twenty-seven SFBB genes,including22 full-length and five nonfull-length SFBB genes,were identified in‘Yali’(P.bretschneideri).SFBBs linkage analysis by PCR-enzyme-linked immunoassay(ELISA)showed that 12 SFBB genes belong to the S21 locus,and 15 SFBB genes belong to the S34 locus.Phylogenetic analysis showed that SFBB genes from Pyrus were divided into 26 types,more than the original eight types.The intrahaplotypic divergence of SFBBs is high and comparable to the allelic diversity of S-RNase,which is consistent with a nonself-recognition SI system.In addition,the expression level of PbrSFBBs in‘Jinzhui’,the only known haploid pollen of a self-compatible mutant,was mostly approximately two times higher than in‘Yali’,which may be the reason for the self-compatible mutant.
基金Supported by National Natural Science Foundation of China(30960200)National Science and Technology Support Program of China(2011BAD35B04)Applied Basic Research Project of Qinghai Province(2011-Z-701)
文摘Brassica rapa cuhivars Qinghaidahuang and Menyuan small rape were used as female parents and separately interspecifically hybridized with Zhongchijie- lan to artificially synthesize a number of Brasica napus resources. Observation of meiosis, pollen fertility and pollen tube germination and analysis of self-compati- bility index of these artificial B. napus varieties were conducted. Results showed that pollen mother cells of artificial B. napus lines from two cross combinations all had normal chromosome behavior and high pollen fertility, but the selfing seed-setting and self-compatibility index varied extremely significantly; B. napus lines from cross combination Qinghaidahuang x Zhongchijidan are highly self-compatible, while B. napus lines from cross combination Menyuan small rape x Zhongchijielan are highly self-incompatible. This study analyzed the heredity laws of self-incompatibility trait in artificially synthesized lines and discussed applica- tion prospects of artificially synthesized self-incompatible B. napus lines in rapeseed production.
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’.
