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1.
Virus Res ; 323: 199000, 2023 Jan 02.
Article in English | MEDLINE | ID: mdl-36356676

ABSTRACT

African swine fever virus (ASFV), a highly pathogenic large DNA virus, is the cause of African swine fever worldwide. The ASFV virulence gene EP402R encodes CD2v, a structural protein that plays an important role in the ASFV infection process. In this study, a CHO-S cell line stably expressing the extracellular region of CD2v was generated and secretory CD2v(sCD2v)was purified from the cell culture supernatant. The purified glycosylated sCD2v protein possessed high immunoreactivity and immunogenicity. In addition, we found that glycosylation had a decisive effect on the immune reactivity of CD2v. Then sCD2v was used to generate five CD2v-specific monoclonal antibodies. The reactivity of all monoclonal antibodies with CD2v protein was confirmed by Western blot and indirect immunofluorescence assay (IFA). Interestingly, mAb 8D5 reactivity with sCD2v depended on sCD2v glycosylation status. Subsequent B cell epitope mapping experiments conducted using a series of overlapping synthetic peptides of the CD2v extracellular domain led to identification of mAb B cell epitopes of 128TCKKNNGTNT137 for mAb 4B11 and 148VKYTNESILE157 for mAbs 5H4 and 5F7. Due to their well-defined epitopes, these three mAbs will likely serve as valuable tools for use in ASFV CD2v structure-function studies, diagnostic assays, and prophylactic methodologies to control ASFV transmission.

2.
Front Plant Sci ; 11: 574982, 2020.
Article in English | MEDLINE | ID: mdl-33193512

ABSTRACT

Apricot mei, a hybrid of Prunus mume and Prunus sibirica, usually has greater cold resistance than P. mume; however, most varieties of Apricot mei lack the characteristic floral scent of P. mume. The volatile and intracellular metabolites, activity levels of key enzymes, and transcriptomes of blooming flowers were comprehensively investigated in five varieties of P. mume. Benzyl acetate and eugenol were determined to be the main components of the P. mume floral scent. However, benzyl benzoate and benzyl alcohol benzoyltransferase activity was detected in only the low-fragrance varieties "Dan Fenghou" and "Yanxing." No benzyl alcohol or benzaldehyde reductase (BAR) activity was detected in the non-fragrant variety "Fenghou." PmBAR1 and PmBAR3 were identified as the key genes responsible for BAR activity. The lack of benzyl alcohol synthesis in the "Fenghou" variety was caused by low activity of PmBAR1-Fen and low expression of PmBAR3. The 60-aa segment at the N-terminus of PmBAR3 was found to play an important role in its enzymatic activity. Correlation tests between floral scent metabolites and the transcriptomes of the five different scented varieties showed that some transcripts associated with hormones, stresses, posttranslational modifications and transporters may also play important regulatory roles in floral scent metabolism in the different varieties.

3.
Mol Biol Rep ; 47(9): 6635-6647, 2020 Sep.
Article in English | MEDLINE | ID: mdl-32803506

ABSTRACT

Prunus sibirica and Prunus mume are closely related plant species that differ in cold tolerance. Hybrids of P. sibirica and true mume, belonging to the apricot mei group, inherited strong cold resistance from P. sibirica. These materials are favourable for research on the molecular mechanisms of cold resistance. However, no suitable reference genes have been identified for analysing gene expression patterns between P. sibirica and P. mume. Ten candidate reference genes were assessed, namely, actins (ACT2-1, ACT2-2, ACT2-3, ACT2-4), protein phosphatase 2A-1 (PP2A-1), ubiquitins (UBQ2, UBQ3), ubiquitin extension protein (UBQ1) and tubulins (TUB1, TUB2), with four distinct algorithms (geNorm, NormFinder, BestKeeper and RefFinder). UBQ2 was recognized as the best reference gene in stems and buds across materials (P. sibirica; 'Xiaohong Zhusha', 'Beijing Yudie', and 'Xiao Lve' for true mume; and 'Dan Fenghou', 'Fenghou', and 'Yanxing' for apricot mei) under cold stress. In addition, the temporal and spatial expression patterns of PmCBF6 and PmLEA10 among seven varieties during winter periods were analysed using UBQ2 as a reference gene. The expression differed significantly among cultivars, which may contribute to their differences in cold tolerance. This paper confirmed the strong cold tolerance of apricot mei. And the best internal reference gene suitable for seven varieties was selected: UBQ2. Based on the above results, the expression of PmCBF6 and PmLEA10 genes during wintering in seven varieties was analysed. The molecular mechanisms of cold resistance were found to be possibly different in different varieties of P. sibirica and P. mume.


Subject(s)
Cold-Shock Response/genetics , Gene Expression Profiling/methods , Genes, Plant , Plant Proteins/genetics , Plant Proteins/metabolism , Prunus/genetics , Prunus/metabolism , Actins/genetics , Actins/metabolism , Algorithms , Phylogeny , Protein Phosphatase 2/genetics , Protein Phosphatase 2/metabolism , Real-Time Polymerase Chain Reaction , Transcriptome , Tubulin/genetics , Tubulin/metabolism , Ubiquitins/genetics , Ubiquitins/metabolism
4.
Genes (Basel) ; 10(10)2019 10 18.
Article in English | MEDLINE | ID: mdl-31635348

ABSTRACT

Rosa chinensis, an important ancestor species of Rosa hybrida, the most popular ornamental plant species worldwide, produces flowers with diverse colors and fragrances. The R2R3-MYB transcription factor family controls a wide variety of plant-specific metabolic processes, especially phenylpropanoid metabolism. Despite their importance for the ornamental value of flowers, the evolution of R2R3-MYB genes in plants has not been comprehensively characterized. In this study, 121 predicted R2R3-MYB gene sequences were identified in the rose genome. Additionally, a phylogenomic synteny network (synnet) was applied for the R2R3-MYB gene families in 35 complete plant genomes. We also analyzed the R2R3-MYB genes regarding their genomic locations, Ka/Ks ratio, encoded conserved motifs, and spatiotemporal expression. Our results indicated that R2R3-MYBs have multiple synteny clusters. The RcMYB114a gene was included in the Rosaceae-specific Cluster 54, with independent evolutionary patterns. On the basis of these results and an analysis of RcMYB114a-overexpressing tobacco leaf samples, we predicted that RcMYB114a functions in the phenylpropanoid pathway. We clarified the relationship between R2R3-MYB gene evolution and function from a new perspective. Our study data may be relevant for elucidating the regulation of floral metabolism in roses at the transcript level.


Subject(s)
Evolution, Molecular , Genome, Plant , Plant Proteins/genetics , Rosa/genetics , Transcription Factors/genetics , Multigene Family , Phylogeny , Plant Proteins/metabolism , Rosa/classification , Synteny , Transcription Factors/metabolism
5.
Front Plant Sci ; 10: 1098, 2019.
Article in English | MEDLINE | ID: mdl-31552079

ABSTRACT

Petal expansion is the main process by which flower opening occurs in roses (Rosa chinensis). Although the regulation of leaf expansion has been extensively studied, little is known about the mechanisms controlling petal expansion. The regulation of leaf dorsoventral (adaxial-abaxial) polarity is important for blade expansion and morphogenesis, but the mechanisms involved adaxial-abaxial regulation in petals are unknown. We found that auxin, a key hormonal regulator of leaf adaxial-abaxial patterning, is unevenly distributed in rose petals. The transcriptomes of the adaxial and abaxial petal tissues were sequenced at three developmental stages during flower opening. Genes that were differentially expressed between the two tissues were filtered for those known to be involved in petal expansion and phytohormone biosynthesis, transport, and signaling, revealing potential roles in petal expansion, especially auxin pathway genes. Using a weighted gene coexpression network analysis (WGCNA), we identified two gene modules that may involve in adaxial-abaxial regulation, 21 and five hub genes have been found respectively. The qRT-PCR validation results were consistent with the RNA-seq data. Based on these findings, we propose a simple network of adaxial-abaxial-related genes that regulates petal expansion in R. chinensis "Old Blush." For the first time, we report the adaxial-abaxial transcriptional changes that occur during petal expansion, providing a reference for the study of the regulation of polarity in plant development.

6.
Int J Mol Sci ; 20(14)2019 Jul 20.
Article in English | MEDLINE | ID: mdl-31330828

ABSTRACT

Rosa chinensis is one of the most popular flower plants worldwide. The recurrent flowering trait greatly enhances the ornamental value of roses, and is the result of the constant formation of new flower buds. Flower bud differentiation has always been a major topic of interest among researchers. The APETALA1 (AP1) MADS-box (Mcm1, Agamous, Deficiens and SRF) transcription factor-encoding gene is important for the formation of the floral meristem and floral organs. However, research on the rose AP1 gene has been limited. Thus, we isolated AP1 from Rosa chinensis 'Old Blush'. An expression analysis revealed that RcAP1 was not expressed before the floral primordia formation stage in flower buds. The overexpression of RcAP1 in Arabidopsis thaliana resulted in an early-flowering phenotype. Additionally, the virus-induced down-regulation of RcAP1 expression delayed flowering in 'Old Blush'. Moreover, RcAP1 was specifically expressed in the sepals of floral organs, while its expression was down-regulated in abnormal sepals and leaf-like organs. These observations suggest that RcAP1 may contribute to rose bud differentiation as well as floral organ morphogenesis, especially the sepals. These results may help for further characterization of the regulatory mechanisms of the recurrent flowering trait in rose.


Subject(s)
Flowers/embryology , Flowers/metabolism , Plant Proteins/metabolism , Rosa/embryology , Rosa/metabolism , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/physiology , Morphogenesis/genetics , Morphogenesis/physiology , Plant Proteins/genetics
7.
Int J Mol Sci ; 19(7)2018 Jun 22.
Article in English | MEDLINE | ID: mdl-29932122

ABSTRACT

Heteromorphic self-incompatibility (SI) is an important system for preventing inbreeding in the genus Primula. However, investigations into the molecular mechanisms of Primula SI are lacking. To explore the mechanisms of SI in Primula maximowiczii, the pollen germination and fruiting rates of self- and cross-pollinations between pin and thrum morphs were investigated, and transcriptomics analyses of the pistils after pollination were performed to assess gene expression patterns in pin and thrum SI. The results indicated that P. maximowiczii exhibits strong SI and that the mechanisms of pollen tube inhibition differ between pin and thrum morphs. While self-pollen tubes of the pin morph were able to occasionally, though rarely, enter the style, those of the thrum morph were never observed to enter the style. The transcriptomics analysis of the pistils revealed 1311 and 1048 differentially expressed genes (DEGs) that were identified by comparing pin self-pollination (PS) vs. pin cross-pollination (PT) and thrum self-pollination (TS) vs. thrum cross-pollination (TP). Notably, about 90% of these DEGs exhibited different expression patterns in the two comparisons. Moreover, pin and thrum DEGs were associated with different Gene Ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways following enrichment analyses. Based on our results, the molecular mechanisms underlying the pin and thrum SI in P. maximowiczii appear to be distinct. Furthermore, the genes involved in the SI processes are commonly associated with carbohydrate metabolism and environmental adaptation. These results provide new insight into the molecular mechanisms of Primula SI.


Subject(s)
Gene Expression Profiling/methods , Gene Expression Regulation, Plant , Primula/genetics , Self-Incompatibility in Flowering Plants/genetics , Flowers/genetics , Gene Ontology , Pollen/genetics , Pollen Tube/genetics , Pollination/genetics
8.
Front Plant Sci ; 9: 481, 2018.
Article in English | MEDLINE | ID: mdl-29706982

ABSTRACT

Rosa chinensis, which is a famous traditional flower in China, is a major ornamental plant worldwide. Long-term cultivation and breeding have resulted in considerable changes in the number of rose petals, while most wild Rosaceae plants have only one whorl consisting of five petals. The petals of double flowers reportedly originate from stamens, but the underlying molecular mechanism has not been fully characterized. In this study, we observed that the number of petals of R. chinensis 'Old Blush' flowers increased and decreased in response to low- and high-temperature treatments, respectively, similar to previous reports. We characterized these variations in further detail and found that the number of stamens exhibited the opposite trend. We cloned an APETALA2 homolog, RcAP2. A detailed analysis of gene structure and promoter cis-acting elements as well as RcAP2 temporospatial expression patterns and responses to temperature changes suggested that RcAP2 expression may be related to the number of petals from stamen origin. The overexpression of RcAP2 in Arabidopsis thaliana transgenic plants may induce the transformation of stamens to petals, thereby increasing the number of petals. Moreover, silencing RcAP2 in 'Old Blush' plants decreased the number of petals. Our results may be useful for clarifying the temperature-responsive mechanism involved in petaloid stamen production, which may be relevant for the breeding of new rose varieties with enhanced flower traits.

9.
Front Plant Sci ; 9: 17, 2018.
Article in English | MEDLINE | ID: mdl-29449849

ABSTRACT

Dormancy Associated MADS-box genes are SVP/MADs-box members and supposed to play crucial roles in plant dormancy of perennial species. In Prunus mume, PmDAM6 has been previously identified to induce plant dormancy. In the current study, six PmDAMs were cloned in P. mume and functionally analyzed in yeast and tobacco to detect the roles of the genes paralogous to PmDAM6. The expression patterns together with sequence similarities indicate that PmDAMs are divided into two sub-clades within SVP group. Moreover, PmDAMs are verified to take part in the development of different plant organs, specifically the flower buds, in some intricate patterns. Furthermore, the PmDAM proteins are found to have special functions by forming corresponding protein complex during the development of flower bud and induction of dormancy. In particular, when PmDAM1 dominating in flower bud in the warm months, the protein complexes are consisted of PmDAM1 itself or with PmDAM2. With the decrease temperatures in the following months, PmDAM6 was found to be highly expressed and gradually changed the complex structure to PmDAM6-protein complex due to strong binding tendencies with PmDAM1 and PmDAM3. Finally, the homodimers of PmDAM6 prevailed to induce the dormancy. The results obtained in the current study highlight the functions of PmDAMs in the tissue development and dormancy, which provide available suggestions for further explorations of protein-complex functions in association with bud growth and dormancy.

10.
Front Plant Sci ; 8: 319, 2017.
Article in English | MEDLINE | ID: mdl-28344586

ABSTRACT

Mei (Prunus mume) is a peculiar woody ornamental plant famous for its inviting fragrance in winter. However, in this valuable plant, the mechanism behind floral volatile development remains poorly defined. Therefore, to explore the floral scent formation, a comparative transcriptome was conducted in order to identify the global transcripts specifying flower buds and blooming flowers of P. mume. Differentially expressed genes were identified between the two different stages showing great discrepancy in floral volatile production. Moreover, according to the expression specificity among the organs (stem, root, fruit, leaf), we summarized one gene cluster regulating the benzenoid floral scent. Significant gene changes were observed in accordance with the formation of benzenoid, thus pointing the pivotal roles of genes as well as cytochrome-P450s and short chain dehydrogenases in the benzenoid biosynthetic process. Further, transcription factors like EMISSION OF BENZENOID I and ODORANT I performed the same expression pattern suggesting key roles in the management of the downstream genes. Taken together, these data provide potential novel anchors for the benzenoid pathway, and the insight for the floral scent induction and regulation mechanism in woody plants.

11.
Sci Rep ; 7: 43382, 2017 02 22.
Article in English | MEDLINE | ID: mdl-28225056

ABSTRACT

The developmental process that produces the ornate petals of the China rose (Rosa chinensis) is complex and is thought to depend on the balanced expression of a functionally diverse array of genes; however, the molecular basis of rose petal development is largely unknown. Here, petal growth of the R. chinensis cultivar 'Old Blush' was divided into four developmental stages, and RNA-seq technology was used to analyse the dynamic changes in transcription that occur as development progresses. In total, 598 million clean reads and 61,456 successfully annotated unigenes were obtained. Differentially expressed gene (DEG) analysis comparing the transcriptomes of the developmental stages resulted in the identification of several potential candidate genes involved in petal development. DEGs involved in anthocyanin biosynthesis, petal expansion, and phytohormone pathways were considered in depth, in addition to several candidate transcription factors. These results lay a foundation for future studies on the regulatory mechanisms underlying rose petal development and may be used in molecular breeding programs aimed at generating ornamental rose lines with desirable traits.


Subject(s)
Flowers/genetics , Gene Expression Regulation, Plant , Genome, Plant , Plant Proteins/genetics , RNA, Plant/genetics , Rosa/genetics , Anthocyanins/biosynthesis , Anthocyanins/genetics , Flowers/growth & development , Flowers/metabolism , Gene Expression Profiling , Gene Expression Regulation, Developmental , Gene Ontology , High-Throughput Nucleotide Sequencing , Molecular Sequence Annotation , Plant Breeding , Plant Growth Regulators/biosynthesis , Plant Growth Regulators/genetics , Plant Proteins/metabolism , RNA, Plant/metabolism , Rosa/growth & development , Rosa/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Transcriptome
12.
Front Plant Sci ; 8: 151, 2017.
Article in English | MEDLINE | ID: mdl-28224001

ABSTRACT

Dehydrins, known as group 2 or D-11 family late-embryogenesis-abundant (LEA) proteins, play important roles in plant growth and stress tolerance. Six dehydrin genes were previously identified from the genome of Prunus mume. In this study, five of them (PmLEA8, PmLEA10, PmLEA19, PmLEA20, and PmLEA29) were cloned from cold-resistant P. mume 'Beijingyudie'. Real-time RT-PCR analysis indicated that all these genes could be up-regulated by one or several treatments (ABA, SA, low temperature, high temperature, PEG, and NaCl treatments). The results of spot assay demonstrated that the expression of all these dehydrins, except PmLEA8, conferred improved osmotic and freezing-resistance to the recombinant Escherichia coli. So four dehydrin genes, PmLEA10, PmLEA19, PmLEA20 and PmLEA29 were chosen for individual over-expression in tobacco plants. The transgenic tobacco plants showed lower relative content of malondialdehyde, relative electrolyte leakage and higher relative content of water than control plants when exposed to cold and drought stress. These results demonstrated that PmLEAs were involved in plant responses to cold and drought.

13.
Plant Mol Biol Report ; 35(1): 37-46, 2017.
Article in English | MEDLINE | ID: mdl-28239231

ABSTRACT

Next-generation sequencing technologies provide opportunities to ascertain the genetic basis of phenotypic differences, even in the closely related cultivars via detection of large amount of DNA polymorphisms. In this study, we performed whole-genome re-sequencing of two mei cultivars with contrasting tree architecture. 75.87 million 100 bp pair-end reads were generated, with 92 % coverage of the genome. Re-sequencing data of two former upright mei cultivars were applied for detecting DNA polymorphisms, since we were more interested in variations conferring weeping trait. Applying stringent parameters, 157,317 mutual single nucleotide polymorphisms (SNPs) and 15,064 mutual insertions-deletions (InDels) were detected and found unevenly distributed within and among the mei chromosomes, which lead to the discovery of 220 high-density, 463 low-density SNP regions together with 80 high-density InDel regions. Additionally, 322 large-effect SNPs and 433 large-effect InDels were detected, and 10.09 % of the SNPs were observed in coding regions. 5.25 % SNPs in coding regions resulted in non-synonymous changes. Ninety SNPs were chosen randomly for validation using high-resolution melt analysis. 93.3 % of the candidate SNPs contained the predicted SNPs. Pfam analysis was further conducted to better understand SNP effects on gene functions. DNA polymorphisms of two known QTL loci conferring weeping trait and their functional effect were also analyzed thoroughly. This study highlights promising functional markers for molecular breeding and a whole-genome genetic basis of weeping trait in mei.

14.
BMC Plant Biol ; 17(1): 10, 2017 01 13.
Article in English | MEDLINE | ID: mdl-28086797

ABSTRACT

BACKGROUND: Flower phylogenetics and genetically controlled development have been revolutionised during the last two decades. However, some of these evolutionary aspects are still debatable. MADS-box genes are known to play essential role in specifying the floral organogenesis and differentiation in numerous model plants like Petunia hybrida, Arabidopsis thaliana and Antirrhinum majus. SEPALLATA (SEP) genes, belonging to the MADS-box gene family, are members of the ABCDE and quartet models of floral organ development and play a vital role in flower development. However, few studies of the genes in Prunus mume have yet been conducted. RESULTS: In this study, we cloned four PmSEPs and investigated their phylogenetic relationship with other species. Expression pattern analyses and yeast two-hybrid assays of these four genes indicated their involvement in the floral organogenesis with PmSEP4 specifically related to specification of the prolificated flowers in P. mume. It was observed that the flower meristem was specified by PmSEP1 and PmSEP4, the sepal by PmSEP1 and PmSEP4, petals by PmSEP2 and PmSEP3, stamens by PmSEP2 and PmSEP3 and pistils by PmSEP2 and PmSEP3. CONCLUSION: With the above in mind, flower development in P. mume might be due to an expression of SEP genes. Our findings can provide a foundation for further investigations of the transcriptional factors governing flower development, their molecular mechanisms and genetic basis.


Subject(s)
Flowers/genetics , Genes, Plant , Prunus/genetics , Cloning, Molecular , Flowers/growth & development , MADS Domain Proteins/genetics , Phylogeny , Plant Proteins/genetics , Protein Binding , Prunus/classification , Prunus/growth & development
15.
Front Plant Sci ; 7: 1301, 2016.
Article in English | MEDLINE | ID: mdl-27630648

ABSTRACT

TCP proteins, belonging to a plant-specific transcription factors family, are known to have great functions in plant development, especially flower and leaf development. However, there is little information about this gene family in Prunus mume, which is widely cultivated in China as an ornamental and fruit tree. Here a genome-wide analysis of TCP genes was performed to explore their evolution in P. mume. Nineteen PmTCPs were identified and three of them contained putative miR319 target sites. Phylogenetic and comprehensive bioinformatics analyses of these genes revealed that different types of TCP genes had undergone different evolutionary processes and the genes in the same clade had similar chromosomal location, gene structure, and conserved domains. Expression analysis of these PmTCPs indicated that there were diverse expression patterns among different clades. Most TCP genes were predominantly expressed in flower, leaf, and stem, and showed high expression levels in the different stages of flower bud differentiation, especially in petal formation stage and gametophyte development. Genes in TCP-P subfamily had main roles in both flower development and gametophyte development. The CIN genes in double petal cultivars might have key roles in the formation of petal, while they were correlated with gametophyte development in the single petal cultivar. The CYC/TB1 type genes were highly detected in the formation of petal and pistil. The less-complex flower types of P. mume might result from the fact that there were only two CYC type genes present in P. mume and a lack of CYC2 genes to control the identity of flower types. These results lay the foundation for further study on the functions of TCP genes during flower development.

16.
Mol Genet Genomics ; 290(5): 1701-15, 2015 Oct.
Article in English | MEDLINE | ID: mdl-25810323

ABSTRACT

SQUAMOSA promoter-binding protein (SBP)-box family genes encode plant-specific transcription factors that play crucial roles in plant development, especially flower and fruit development. However, little information on this gene family is available for Prunus mume, an ornamental and fruit tree widely cultivated in East Asia. To explore the evolution of SBP-box genes in Prunus and explore their functions in flower and fruit development, we performed a genome-wide analysis of the SBP-box gene family in P. mume. Fifteen SBP-box genes were identified, and 11 of them contained an miR156 target site. Phylogenetic and comprehensive bioinformatics analyses revealed that different groups of SBP-box genes have undergone different evolutionary processes and varied in their length, structure, and motif composition. Purifying selection has been the main selective constraint on both paralogous and orthologous SBP-box genes. In addition, the sequences of orthologous SBP-box genes did not diverge widely after the split of P. mume and Prunus persica. Expression analysis of P. mume SBP-box genes revealed their diverse spatiotemporal expression patterns. Three duplicated SBP-box genes may have undergone subfunctionalization in Prunus. Most of the SBP-box genes showed high transcript levels in flower buds and young fruit. The four miR156-nontargeted genes were upregulated during fruit ripening. Together, these results provide information about the evolution of SBP-box genes in Prunus. The expression analysis lays the foundation for further research on the functions of SBP-box genes in P. mume and other Prunus species, especially during flower and fruit development.


Subject(s)
Gene Expression Profiling , Genes, Plant , Plant Proteins/genetics , Prunus/genetics , Amino Acid Sequence , Base Sequence , Chromosome Mapping , Chromosomes, Plant , MicroRNAs/genetics , Molecular Sequence Data , Phylogeny , Plant Proteins/chemistry , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid
17.
DNA Res ; 22(3): 183-91, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25776277

ABSTRACT

High-density genetic map is a valuable tool for fine mapping locus controlling a specific trait especially for perennial woody plants. In this study, we firstly constructed a high-density genetic map of mei (Prunus mume) using SLAF markers, developed by specific locus amplified fragment sequencing (SLAF-seq). The linkage map contains 8,007 markers, with a mean marker distance of 0.195 cM, making it the densest genetic map for the genus Prunus. Though weeping trees are used worldwide as landscape plants, little is known about weeping controlling gene(s) (Pl). To test the utility of the high-density genetic map, we did fine-scale mapping of this important ornamental trait. In total, three statistic methods were performed progressively based on the result of inheritance analysis. Quantitative trait loci (QTL) analysis initially revealed that a locus on linkage group 7 was strongly responsible for weeping trait. Mutmap-like strategy and extreme linkage analysis were then applied to fine map this locus within 1.14 cM. Bioinformatics analysis of the locus identified some candidate genes. The successful localization of weeping trait strongly indicates that the high-density map constructed using SLAF markers is a worthy reference for mapping important traits for woody plants.


Subject(s)
Chromosome Mapping , Chromosomes, Plant , Prunus/genetics , Quantitative Trait Loci , Genetic Linkage , Genotype , Sequence Analysis, DNA
18.
Biosci Biotechnol Biochem ; 78(10): 1640-7, 2014.
Article in English | MEDLINE | ID: mdl-25273130

ABSTRACT

In order to investigate the difference in their characteristic floral scents between Prunus mume Siebold & Zucc. and the related Prunus species, their headspace volatiles and endogenous extraction were analyzed by gas chromatography-mass spectrometry. The efficiency of substrate utilization of the flowers was studied by incubating them with different alcohol substrates. Our results indicated that benzyl acetate is a dominant compound influencing the characteristic floral scent of P. mume. An alcohol substrate concentration of 4 mmol L(-1) and a reaction time of 2 h were constituted the reaction condition for catalysis of exogenous alcohol substrates by the flowers. Under these conditions, Prunus sibirica exhibited the highest utilization efficiency for benzyl alcohol substrate while the utilization efficiency of Prunus persica was the lowest. Comparative analysis of several alcohol substrates indicated that the flowers of the tested species had selective specificity for benzyl alcohol substrates.


Subject(s)
Flowers/chemistry , Odorants/analysis , Prunus/chemistry , Volatile Organic Compounds/analysis , Benzyl Alcohol/pharmacology , Breeding , Flowers/drug effects , Prunus/drug effects , Species Specificity
19.
BMC Genet ; 15 Suppl 1: S1, 2014.
Article in English | MEDLINE | ID: mdl-25078672

ABSTRACT

Mei, Prunus mume Sieb. et Zucc., is an ornamental plant popular in East Asia and, as an important member of genus Prunus, has played a pivotal role in systematic studies of the Rosaceae. However, the genetic architecture of botanical traits in this species remains elusive. This paper represents the first genome-wide mapping study of quantitative trait loci (QTLs) that affect stem growth and form, leaf morphology and leaf anatomy in an intraspecific cross derived from two different mei cultivars. Genetic mapping based on a high-density linkage map constricted from 120 SSRs and 1,484 SNPs led to the detection of multiple QTLs for each trait, some of which exert pleiotropic effects on correlative traits. Each QTL explains 3-12% of the phenotypic variance. Several leaf size traits were found to share common QTLs, whereas growth-related traits and plant form traits might be controlled by a different set of QTLs. Our findings provide unique insights into the genetic control of tree growth and architecture in mei and help to develop an efficient breeding program for selecting superior mei cultivars.


Subject(s)
Chromosome Mapping , Genetic Linkage , Prunus/anatomy & histology , Prunus/genetics , DNA, Plant/genetics , Microsatellite Repeats , Phenotype , Polymorphism, Single Nucleotide , Prunus/growth & development , Quantitative Trait Loci
20.
Mol Genet Genomics ; 289(5): 903-20, 2014 Oct.
Article in English | MEDLINE | ID: mdl-24859011

ABSTRACT

MADS-box genes encode transcription factors that play crucial roles in plant development, especially in flower and fruit development. To gain insight into this gene family in Prunus mume, an important ornamental and fruit plant in East Asia, and to elucidate their roles in flower organ determination and fruit development, we performed a genome-wide identification, characterisation and expression analysis of MADS-box genes in this Rosaceae tree. In this study, 80 MADS-box genes were identified in P. mume and categorised into MIKC, Mα, Mß, Mγ and Mδ groups based on gene structures and phylogenetic relationships. The MIKC group could be further classified into 12 subfamilies. The FLC subfamily was absent in P. mume and the six tandemly arranged DAM genes might experience a species-specific evolution process in P. mume. The MADS-box gene family might experience an evolution process from MIKC genes to Mδ genes to Mα, Mß and Mγ genes. The expression analysis suggests that P. mume MADS-box genes have diverse functions in P. mume development and the functions of duplicated genes diverged after the duplication events. In addition to its involvement in the development of female gametophytes, type I genes also play roles in male gametophytes development. In conclusion, this study adds to our understanding of the roles that the MADS-box genes played in flower and fruit development and lays a foundation for selecting candidate genes for functional studies in P. mume and other species. Furthermore, this study also provides a basis to study the evolution of the MADS-box family.


Subject(s)
Genome, Plant , MADS Domain Proteins/genetics , Plant Proteins/genetics , Prunus/genetics , Amino Acid Motifs , Chromosome Mapping , Conserved Sequence , Evolution, Molecular , Flowers/genetics , Flowers/growth & development , Flowers/metabolism , Fruit/genetics , Fruit/growth & development , Fruit/metabolism , Gene Expression , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , MADS Domain Proteins/chemistry , MADS Domain Proteins/metabolism , Organ Specificity , Phylogeny , Plant Proteins/chemistry , Plant Proteins/metabolism , Prunus/growth & development , Prunus/metabolism
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