Long-read bitter gourd (Momordica charantia) genome and the genomic architecture of nonclassic domestication.

The genetic architecture of quantitative traits is determined by both Mendelian and polygenic factors, yet classic examples of plant domestication focused on selective sweep of newly mutated Mendelian genes. Here we report the chromosome-level genome assembly and the genomic investigation of a nonclassic domestication example, bitter gourd (Momordica charantia), an important Asian vegetable and medicinal plant of the family Cucurbitaceae. Population resequencing revealed the divergence between wild and South Asian cultivars about 6,000 y ago, followed by the separation of the Southeast Asian cultivars about 800 y ago, with the latter exhibiting more extreme trait divergence from wild progenitors and stronger signs of selection on fruit traits. Unlike some crops where the largest phenotypic changes and traces of selection happened between wild and cultivar groups, in bitter gourd large differences exist between two regional cultivar groups, likely reflecting the distinct consumer preferences in different countries. Despite breeding efforts toward increasing female flower proportion, a gynoecy locus exhibits complex patterns of balanced polymorphism among haplogroups, with potential signs of selective sweep within haplogroups likely reflecting artificial selection and introgression from cultivars back to wild accessions. Our study highlights the importance to investigate such nonclassic example of domestication showing signs of balancing selection and polygenic trait architecture in addition to classic selective sweep in Mendelian factors.

Genomic characterization of a rare Carica papaya X chromosome mutant reveals a candidate monodehydroascorbate reductase 4 gene involved in all-hermaphrodite phenomenon.

Sex form is one of the most important characteristics in papaya cultivation in which hermaphrodite is the preferable form. Self-pollination of H*-TSS No.7, an inbred line derived from a rare X chromosome mutant SR*, produced all-hermaphrodite progeny. The recessive lethal allele controlling the all-hermaphrodite phenomenon was proposed to be the recessive Germination suppressor (gs) locus. This study employed next-generation sequencing technology and genome comparison to identify the candidate Gs gene. One specific gene, monodehydroascorbate reductase 4 (MDAR4) harboring a unique polymorphic 3 bp deletion in H*-TSS No.7 was identified. The function of MDAR4 is known to be involved in the hydrogen peroxide (H2O2) scavenging pathway and is associated with seed germination. Furthermore, MDAR4 showed higher expression in the imbibed seeds than that in the dry seeds indicating its potential role in the seed germination. Perhaps this is the very first report providing the evidences that MDAR4 is the candidate of Gs locus in H*-TSS No.7. In addition, Gs allele-specific markers were developed which would be facilitated for breeding all-hermaphrodite lines.

Shear bond strength between gingival composite resin and glazed gingival porcelain for implant-supported prostheses.

This study determined the shear bond strength (SBS) between an indirect gingival composite resin and glazed gingival porcelain after various surface treatments. A total of 176 porcelain disks with natural glazing were used and assigned to one of four groups: no surface treatment, airborne-particle abrasion, hydrofluoric acid etching, or a combination of airborne-particle abrasion followed by hydrofluoric acid etching. Each group was divided into two subgroups: one subgroup was unprimed, and the other was silanized. An indirect composite resin was then bonded to the porcelain disks. Half of the specimens in each group (n = 11) were exposed to 5000 thermocycles. SBSs were measured, and data were analyzed with the Kruskal-Wallis and Steel-Dwass tests. Among silanized specimens, those treated with the combination of airborne-particle abrasion and hydrofluoric acid etching exhibited the highest bond strengths both before and after thermocycling. However, the SBS values of the silanized and unprimed hydrofluoric acid etched specimens did not differ significantly. Airborne-particle abrasion followed by hydrofluoric acid etching with silane application yielded stronger, more durable bonds between the indirect gingival composite resin and glazed gingival porcelain.

Radiopacity of computer-aided design/computer-aided manufacturing composite resin blocks.

This study aimed to clarify the degree of difference in radiopacity between sixteen CAD/CAM composite resin blocks, one ceramic block, and teeth of the same thickness on radiographs. The radiographic density of CAD/CAM composite resin blocks was measured and the results were compared with the corresponding values for enamel and dentin. Additionally, the study analyzed the constituent elements of each type of CAD/CAM composite resin block and conducted an examination to identify those elements exerting an influence on radiopacity. Compared to the enamel, there were five blocks with significantly higher radiopacity, two blocks with the same level in radiopacity, and ten blocks with notably lower radiopacity. Compared to the dentin, there were ten blocks with significantly higher radiopacity, one block with the same level in radiopacity, and six blocks with notably lower radiopacity. All of the CAD/CAM composite resin blocks for molars contained barium and strontium. This result suggests that the addition of heavy metals, for example, barium, strontium, and zirconium, would be effective in providing CAD/CAM composite resin blocks with radiopacity.

OsHKT1;5 mediates Na+ exclusion in the vasculature to protect leaf blades and reproductive tissues from salt toxicity in rice.

Salt tolerance quantitative trait loci analysis of rice has revealed that the SKC1 locus, which is involved in a higher K+ /Na+ ratio in shoots, corresponds to the OsHKT1;5 gene encoding a Na+ -selective transporter. However, physiological roles of OsHKT1;5 in rice exposed to salt stress remain elusive, and no OsHKT1;5 gene disruption mutants have been characterized to date. In this study, we dissected two independent T-DNA insertional OsHKT1;5 mutants. Measurements of ion contents in tissues and 22 Na+ tracer imaging experiments showed that loss-of-function of OsHKT1;5 in salt-stressed rice roots triggers massive Na+ accumulation in shoots. Salt stress-induced increases in the OsHKT1;5 transcript were observed in roots and basal stems, including basal nodes. Immuno-staining using an anti-OsHKT1;5 peptide antibody indicated that OsHKT1;5 is localized in cells adjacent to the xylem in roots. Additionally, direct introduction of 22 Na+ tracer to leaf sheaths also demonstrated the involvement of OsHKT1;5 in xylem Na+ unloading in leaf sheaths. Furthermore, OsHKT1;5 was indicated to be present in the plasma membrane and found to localize also in the phloem of diffuse vascular bundles in basal nodes. Together with the characteristic 22 Na+ allocation in the blade of the developing immature leaf in the mutants, these results suggest a novel function of OsHKT1;5 in mediating Na+ exclusion in the phloem to prevent Na+ transfer to young leaf blades. Our findings further demonstrate that the function of OsHKT1;5 is crucial over growth stages of rice, including the protection of the next generation seeds as well as of vital leaf blades under salt stress.

Magnaporthe oryzae Glycine-Rich Secretion Protein, Rbf1 Critically Participates in Pathogenicity through the Focal Formation of the Biotrophic Interfacial Complex.

Magnaporthe oryzae, the fungus causing rice blast disease, should contend with host innate immunity to develop invasive hyphae (IH) within living host cells. However, molecular strategies to establish the biotrophic interactions are largely unknown. Here, we report the biological function of a M. oryzae-specific gene, Required-for-Focal-BIC-Formation 1 (RBF1). RBF1 expression was induced in appressoria and IH only when the fungus was inoculated to living plant tissues. Long-term successive imaging of live cell fluorescence revealed that the expression of RBF1 was upregulated each time the fungus crossed a host cell wall. Like other symplastic effector proteins of the rice blast fungus, Rbf1 accumulated in the biotrophic interfacial complex (BIC) and was translocated into the rice cytoplasm. RBF1-knockout mutants (Δrbf1) were severely deficient in their virulence to rice leaves, but were capable of proliferating in abscisic acid-treated or salicylic acid-deficient rice plants. In rice leaves, Δrbf1 inoculation caused necrosis and induced defense-related gene expression, which led to a higher level of diterpenoid phytoalexin accumulation than the wild-type fungus did. Δrbf1 showed unusual differentiation of IH and dispersal of the normally BIC-focused effectors around the short primary hypha and the first bulbous cell. In the Δrbf1-invaded cells, symplastic effectors were still translocated into rice cells but with a lower efficiency. These data indicate that RBF1 is a virulence gene essential for the focal BIC formation, which is critical for the rice blast fungus to suppress host immune responses.

An in vitro evaluation of fracture load of implant-supported zirconia-based prostheses fabricated with different veneer materials.

OBJECTIVES: The purpose of this in vitro study was to evaluate fracture loads of implant-supported zirconia-based prostheses fabricated with different veneer materials (resin-based material and lithium disilicate ceramics). MATERIAL AND METHODS: Forty-four zirconia-based molar prostheses were fabricated on dental implants and divided into four groups (n = 11): zirconia-based prostheses veneered with feldspathic porcelain (ZVF), zirconia-based prostheses bonded with the lithium disilicate glass-ceramic veneer (ZBD), zirconia-based prostheses veneered with indirect composite resin (ZVC), and zirconia-based prostheses bonded with composite materials fabricated from a CAD/CAM resin block (ZBC). The zirconia-based prostheses and abutments were adhesively bonded with a dual-polymerized resin-based luting material. Fracture load was determined using compression load to the prostheses with a universal testing machine. The data were analyzed with one-way analysis of variance (ANOVA) and Tukey's HSD test (α = .05). RESULTS: The mean fracture load was significantly higher in the ZBC group (3.95 kN) than in the ZVC group (3.28 kN). No significant difference in fracture load was found among the ZVF (3.52 kN), ZBD (3.48 kN), and ZVC groups. CONCLUSIONS: The adhesively bonded veneering technique enhances fracture resistance of implant-supported zirconia-based prostheses fabricated with a resin-based material. All implant-supported zirconia-based restorations tested should resist physiologic masticatory forces in the oral environment.

Effect of preparation design on marginal and internal adaptation of translucent zirconia laminate veneers.

This study investigated the effect of preparation design on the marginal and internal adaptation of laminate veneers (LVs) fabricated from translucent zirconia. Thirty-three resin teeth were prepared for LVs of three designs: window preparation (WP); incisal shoulder preparation (ISP); and incisal palatal chamfer preparation (IPP). Marginal adaptation was evaluated by measuring the vertical discrepancy between the LV margin and the finish line at 60 points. The internal adaptation was assessed by measuring the internal space width as the distance between the LV and the tooth at cervical, central, and incisal sites after sectioning. At the incisal, mesial, and distal sites, mean marginal discrepancies were significantly lower in the WP group than in the other two groups; the IPP group had the highest marginal discrepancies. At incisal sites, the median internal space was significantly higher in the IPP group than in the WP and ISP groups and higher in the ISP group than in the WP group. At the incisal site, marginal and internal adaptations were better for the non-overlap translucent zirconia LV design (WP) than for the overlap designs (ISP and IPP). The characteristics of the translucent zirconia LVs used in the present study indicate acceptable clinical performance.

Genome sequencing of the staple food crop white Guinea yam enables the development of a molecular marker for sex determination.

BACKGROUND: Root and tuber crops are a major food source in tropical Africa. Among these crops are several species in the monocotyledonous genus Dioscorea collectively known as yam, a staple tuber crop that contributes enormously to the subsistence and socio-cultural lives of millions of people, principally in West and Central Africa. Yam cultivation is constrained by several factors, and yam can be considered a neglected "orphan" crop that would benefit from crop improvement efforts. However, the lack of genetic and genomic tools has impeded the improvement of this staple crop. RESULTS: To accelerate marker-assisted breeding of yam, we performed genome analysis of white Guinea yam (Dioscorea rotundata) and assembled a 594-Mb genome, 76.4% of which was distributed among 21 linkage groups. In total, we predicted 26,198 genes. Phylogenetic analyses with 2381 conserved genes revealed that Dioscorea is a unique lineage of monocotyledons distinct from the Poales (rice), Arecales (palm), and Zingiberales (banana). The entire Dioscorea genus is characterized by the occurrence of separate male and female plants (dioecy), a feature that has limited efficient yam breeding. To infer the genetics of sex determination, we performed whole-genome resequencing of bulked segregants (quantitative trait locus sequencing [QTL-seq]) in F1 progeny segregating for male and female plants and identified a genomic region associated with female heterogametic (male = ZZ, female = ZW) sex determination. We further delineated the W locus and used it to develop a molecular marker for sex identification of Guinea yam plants at the seedling stage. CONCLUSIONS: Guinea yam belongs to a unique and highly differentiated clade of monocotyledons. The genome analyses and sex-linked marker development performed in this study should greatly accelerate marker-assisted breeding of Guinea yam. In addition, our QTL-seq approach can be utilized in genetic studies of other outcrossing crops and organisms with highly heterozygous genomes. Genomic analysis of orphan crops such as yam promotes efforts to improve food security and the sustainability of tropical agriculture.

Fracture resistance of implant-supported screw-retained zirconia-based molar restorations.

OBJECTIVES: The objective of this in vitro study was to investigate fracture loads of screw-retained zirconia-based molar restorations (hybrid abutment crown) fabricated with different restorative materials and designs. MATERIAL AND METHODS: Forty-four screw-retained zirconia-based molar restorations were fabricated on dental implants and divided into four groups (n = 11): porcelain-layered zirconia-based restorations (PLZ), indirect composite-layered zirconia-based restorations (ILZ), metal-ceramic restorations (MC), and monolithic zirconia restorations (MONO). The zirconia-based restorations in the PLZ, ILZ, and MONO groups were adhesively bonded on implant abutments with a dual-polymerized resin material. All restorations were tightened on implant bodies with titanium screws and were tested for fracture resistance. The Kruskal-Wallis test and Steel-Dwass test were used to evaluate differences in fracture loads (α = 0.05). RESULTS: As compared with the other groups, the MONO specimens had a significantly higher mean fracture resistance (7.54 kN); no significant differences were found among the PLZ (1.96 kN), ILZ (1.80 kN), and MC (1.45 kN) groups (P > 0.05). For the PLZ, ILZ, and MC groups, all specimens fractured within the layering materials. In contrast, the fracture mode for the MONO group was complete fracture of the restorations. CONCLUSIONS: All restorations withstood the masticatory forces. Fracture loads were significantly higher for screw-retained implant-supported monolithic zirconia restorations than for screw-retained bilayered restorations. For the screw-retained bilayered zirconia-based restorations, the fracture resistance of ILZ restorations was comparable to that of PLZ restorations and MC restorations.

Development of plants resistant to Papaya leaf distortion mosaic virus by intergeneric hybridization between Carica papaya and Vasconcellea cundinamarcensis.

In this study, we confirmed that Vasconcellea cundinamarcensis resists Papaya leaf distortion mosaic virus (PLDMV), and used it to produce intergeneric hybrids with Carica papaya. From the cross between C. papaya and V. cundinamarcensis, we obtained 147 seeds with embryos. Though C. papaya is a monoembryonic plant, multiple embryos were observed in all 147 seeds. We produced 218 plants from 28 seeds by means of embryo-rescue culture. All plants had pubescence on their petioles and stems characteristic of V. cundinamarcensis. Flow cytometry and PCR of 28 plants confirmed they were intergeneric hybrids. To evaluate virus resistance, mechanical inoculation of PLDMV was carried out. The test showed that 41 of 134 intergeneric hybrid plants showed no symptoms and were resistant. The remaining 93 hybrids showed necrotic lesions on the younger leaves than the inoculated leaves. In most of the 93 hybrids, the necrotic lesions enclosed the virus and prevented further spread. These results suggest that the intergeneric hybrids will be valuable material for PLDMV-resistant papaya breeding.

A cytochrome P450, OsDSS1, is involved in growth and drought stress responses in rice (Oryza sativa L.).

Cytochrome P450s are among the largest protein coding gene families in plant genomes. However, majority of the genes remain uncharacterized. Here, we report the characterization of dss1, a rice mutant showing dwarfism and reduced grain size. The dss1 phenotype is caused by a non-synonymous point mutation we identified in DSS1, which is member of a P450 gene cluster located on rice chromosome 3 and corresponds to the previously reported CYP96B4/SD37 gene. Phenotypes of several dwarf mutants characterized in rice are associated with defects in the biosynthesis or perception of the phytohormones gibberellins (GAs) and brassinosteroids (BRs). However, both GA and BR failed to rescue the dss1 phenotype. Hormone profiling revealed the accumulation of abscisic acid (ABA) and ABA metabolites, as well as significant reductions in GA19 and GA53 levels, precursors of the bioactive GA1, in the mutant. The dss1 contents of cytokinin and auxins were not significantly different from wild-type plants. Consistent with the accumulation of ABA and metabolites, germination and early growth was delayed in dss1, which also exhibited an enhanced tolerance to drought. Additionally, expressions of members of the DSS1/CYP96B gene cluster were regulated by drought stress and exogenous ABA. RNA-seq-based transcriptome profiling revealed, among others, that cell wall-related genes and genes involved in lipid metabolism were up- and down-regulated in dss1, respectively. Taken together, these findings suggest that DSS1 mediates growth and stress responses in rice by fine-tuning GA-to-ABA balance, and might as well play a role in lipid metabolism.

Genome sequence comparison reveals a candidate gene involved in male-hermaphrodite differentiation in papaya (Carica papaya) trees.

The sex type of papaya (Carica papaya) is determined by the pair of sex chromosomes (XX, female; XY, male; and XY(h), hermaphrodite), in which there is a non-recombining genomic region in the Y and Y(h) chromosomes. This region is presumed to be involved in determination of males and hermaphrodites; it is designated as the male-specific region in the Y chromosome (MSY) and the hermaphrodite-specific region in the Y(h) chromosome (HSY). Here, we identified the genes determining male and hermaphrodite sex types by comparing MSY and HSY genomic sequences. In the MSY and HSY genomic regions, we identified 14,528 nucleotide substitutions and 965 short indels with a large gap and two highly diverged regions. In the predicted genes expressed in flower buds, we found no nucleotide differences leading to amino acid changes between the MSY and HSY. However, we found an HSY-specific transposon insertion in a gene (SVP like) showing a similarity to the Short Vegetative Phase (SVP) gene. Study of SVP-like transcripts revealed that the MSY allele encoded an intact protein, while the HSY allele encoded a truncated protein. Our findings demonstrated that the SVP-like gene is a candidate gene for male-hermaphrodite determination in papaya.

Effects of framework design and layering material on fracture strength of implant-supported zirconia-based molar crowns.

OBJECTIVES: To evaluate the effects of framework design and layering material on the fracture strength of implant-supported zirconia-based molar crowns. MATERIAL AND METHODS: Sixty-six titanium abutments (GingiHue Post) were tightened onto dental implants (Implant Lab Analog). These abutment-implant complexes were randomly divided into three groups (n = 22) according to the design of the zirconia framework (Katana), namely, uniform-thickness (UNI), anatomic (ANA), and supported anatomic (SUP) designs. The specimens in each design group were further divided into two subgroups (n = 11): zirconia-based all-ceramic restorations (ZAC group) and zirconia-based restorations with an indirect composite material (Estenia C&B) layered onto the zirconia framework (ZIC group). All crowns were cemented on implant abutments, after which the specimens were tested for fracture resistance. The data were analyzed with the Kruskal-Wallis test and the Mann-Whitney U-test with the Bonferroni correction (α = 0.05). RESULTS: The following mean fracture strength values (kN) were obtained in UNI design, ANA design, and SUP design, respectively: Group ZAC, 3.78, 6.01, 6.50 and Group ZIC, 3.15, 5.65, 5.83. In both the ZAC and ZIC groups, fracture strength was significantly lower for the UNI design than the other two framework designs (P = 0.001). Fracture strength did not significantly differ (P > 0.420) between identical framework designs in the ZAC and ZIC groups. CONCLUSIONS: A framework design with standardized layer thickness and adequate support of veneer by zirconia frameworks, as in the ANA and SUP designs, increases fracture resistance in implant-supported zirconia-based restorations under conditions of chewing attrition. Indirect composite material and porcelain perform similarly as layering materials on zirconia frameworks.

Leaf margin phenotype-specific restriction-site-associated DNA-derived markers for pineapple (Ananas comosus L.).

To explore genome-wide DNA polymorphisms and identify DNA markers for leaf margin phenotypes, a restriction-site-associated DNA sequencing analysis was employed to analyze three bulked DNAs of F1 progeny from a cross between a 'piping-leaf-type' cultivar, 'Yugafu', and a 'spiny-tip-leaf-type' variety, 'Yonekura'. The parents were both Ananas comosus var. comosus. From the analysis, piping-leaf and spiny-tip-leaf gene-specific restriction-site-associated DNA sequencing tags were obtained and designated as PLSTs and STLSTs, respectively. The five PLSTs and two STSLTs were successfully converted to cleaved amplified polymorphic sequence (CAPS) or simple sequence repeat (SSR) markers using the sequence differences between alleles. Based on the genotyping of the F1 with two SSR and three CAPS markers, the five PLST markers were mapped in the vicinity of the P locus, with the closest marker, PLST1_SSR, being located 1.5 cM from the P locus. The two CAPS markers from STLST1 and STLST3 perfectly assessed the 'spiny-leaf type' as homozygotes of the recessive s allele of the S gene. The recombination value between the S locus and STLST loci was 2.4, and STLSTs were located 2.2 cM from the S locus. SSR and CAPS markers are applicable to marker-assisted selection of leaf margin phenotypes in pineapple breeding.

Long-term Bond Strength between Layering Indirect Composite Material and Zirconia Coated with Silicabased Ceramics.

PURPOSE: This study evaluated the long-term shear bond strength between an indirect composite material and a zirconia framework coated with silica-based ceramics, taking the effect of different primers into account. MATERIALS AND METHODS: A total of 165 airborne-particle abraded zirconia disks were subjected to one of three pretreatments: no pretreatment (ZR-AB), airborne-particle abrasion of zirconia coated with feldspathic porcelain (ZR-PO-AB), and 9.5% hydrofluoric acid etching of zirconia coated with feldspathic porcelain (ZR-PO-HF). An indirect composite material (Estenia C&B) was then bonded to the zirconia disks after they were treated with one of the following primers: Clearfil Photo Bond (CPB), Clearfil Photo Bond with Clearfil Porcelain Bond Activator (CPB + Activator), Estenia Opaque Primer (EOP), Porcelain Liner M Liquid B (PLB), or no priming (CON, control group). Shear bond strength was tested after 100,000 thermocycles, and the data were analyzed using the Steel-Dwass U-test (α = 0.05). RESULTS: For ZR-PO-AB and ZR-PO-HF specimens, bond strength was highest in the CPB+Activator group (25.8 MPa and 22.4 MPa, respectively). Bond strengths were significantly lower for ZR-AB specimens in the CON and PLB groups and for ZR-PO-AB specimens in the CON, CPB, and EOP groups. CONCLUSION: Combined application of a hydrophobic phosphate monomer (MDP) and silane coupling agent enhanced the long-term bond strength of indirect composite material to a zirconia coated with silica-based ceramics.

Large-scale gene disruption in Magnaporthe oryzae identifies MC69, a secreted protein required for infection by monocot and dicot fungal pathogens.

To search for virulence effector genes of the rice blast fungus, Magnaporthe oryzae, we carried out a large-scale targeted disruption of genes for 78 putative secreted proteins that are expressed during the early stages of infection of M. oryzae. Disruption of the majority of genes did not affect growth, conidiation, or pathogenicity of M. oryzae. One exception was the gene MC69. The mc69 mutant showed a severe reduction in blast symptoms on rice and barley, indicating the importance of MC69 for pathogenicity of M. oryzae. The mc69 mutant did not exhibit changes in saprophytic growth and conidiation. Microscopic analysis of infection behavior in the mc69 mutant revealed that MC69 is dispensable for appressorium formation. However, mc69 mutant failed to develop invasive hyphae after appressorium formation in rice leaf sheath, indicating a critical role of MC69 in interaction with host plants. MC69 encodes a hypothetical 54 amino acids protein with a signal peptide. Live-cell imaging suggested that fluorescently labeled MC69 was not translocated into rice cytoplasm. Site-directed mutagenesis of two conserved cysteine residues (Cys36 and Cys46) in the mature MC69 impaired function of MC69 without affecting its secretion, suggesting the importance of the disulfide bond in MC69 pathogenicity function. Furthermore, deletion of the MC69 orthologous gene reduced pathogenicity of the cucumber anthracnose fungus Colletotrichum orbiculare on both cucumber and Nicotiana benthamiana leaves. We conclude that MC69 is a secreted pathogenicity protein commonly required for infection of two different plant pathogenic fungi, M. oryzae and C. orbiculare pathogenic on monocot and dicot plants, respectively.

Functional characterization of a new grapevine MYB transcription factor and regulation of proanthocyanidin biosynthesis in grapes.

A new regulator of proanthocyanidin (PA) biosynthesis in grapes was found by screening genes coordinately expressed with PA accumulation under different light conditions using a substantially improved method of serial analysis of gene expression (SuperSAGE). This R2R3-MYB transcription factor, VvMYBPAR, shows high protein sequence similarity with PA biosynthesis-regulating plant MYBs, such as VvMYBPA2 and TRANSPARENT TESTA2. Its transcript levels were relatively high in the skins of young berries, whereas the levels were higher in the seeds and at a maximum around veraison. In addition to its response to modified light conditions, the gene responded to abscisic acid application in the skins of cultured berries. Among the PA-specific branch genes, this transcript profile was not correlated with that of VvANR and VvLAR1 but was closely related to that of VvLAR2, suggesting different regulation of PA-specific branch genes from that of a known PA regulator, VvMYBPA2. The PA-specific regulation of VvMYBPAR was confirmed by VvMYBPAR constitutive expression in Arabidopsis in which the transgene specifically induced PA biosynthetic genes and resulted in PA accumulation in plants grown on sucrose-supplemented media to induce anthocyanin synthesis. A transient reporter assay using grapevine cells showed that VvMYBPAR activated the promoters on PA-specific branch genes and candidate genes associated with modification and transport of monomeric PA precursors, as well as the promoters of VvCHS3 and VvF3'5'Hd in the common flavonoid pathway, but not that of VvUFGT on the anthocyanin-specific branch. This new factor suggests the polygenic regulation of PA biosynthesis in grapes by closely related MYB transcription factors.

Rice alcohol dehydrogenase 1 promotes survival and has a major impact on carbohydrate metabolism in the embryo and endosperm when seeds are germinated in partially oxygenated water.

BACKGROUND AND AIMS: Rice (Oryza sativa) has the rare ability to germinate and elongate a coleoptile under oxygen-deficient conditions, which include both hypoxia and anoxia. It has previously been shown that ALCOHOL DEHYDROGENASE 1 (ADH1) is required for cell division and cell elongation in the coleoptile of submerged rice seedlings by means of studies using a rice ADH1-deficient mutant, reduced adh activity (rad). The aim of this study was to understand how low ADH1 in rice affects carbohydrate metabolism in the embryo and endosperm, and lactate and alanine synthesis in the embryo during germination and subsequent coleoptile growth in submerged seedlings. METHODS: Wild-type and rad mutant rice seeds were germinated and grown under complete submergence. At 1, 3, 5 and 7 d after imbibition, the embryo and endosperm were separated and several of their metabolites were measured and compared. KEY RESULTS: In the rad embryo, the rate of ethanol fermentation was halved, while lactate and alanine concentrations were 2·4- and 5·7- fold higher in the mutant than in the wild type. Glucose and fructose concentrations in the embryos increased with time in the wild type, but not in the rad mutant. The rad mutant endosperm had lower amounts of the α-amylases RAMY1A and RAMY3D, resulting in less starch degradation and lower glucose concentrations. CONCLUSIONS: These results suggest that ADH1 is essential for sugar metabolism via glycolysis to ethanol fermentation in both the embryo and endosperm. In the endosperm, energy is presumably needed for synthesis of the amylases and for sucrose synthesis in the endosperm, as well as for sugar transport to the embryo.