• The Plant Pathology Journal
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• v.33 no.2
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• pp.193-205
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• 2017

Histone methylation plays important roles in regulating chromatin dynamics and transcription in eukaryotes. Implication of histone modifications in fungal pathogenesis is, however, beginning to emerge. Here, we report identification and functional analysis of a putative JmjC-domain-containing histone demethylase in Magnaporthe oryzae. Through bioinformatics analysis, we identified seven genes, which encode putative histone demethylases containing JmjC domain. Deletion of one gene, MoJMJ1, belonging to JARID group, resulted in defects in vegetative growth, asexual reproduction, appressorium formation as well as invasive growth in the fungus. Western blot analysis showed that global H3K4me3 level increased in the deletion mutant, compared to wild-type strain, indicating histone demethylase activity of MoJMJ1. Introduction of MoJMJ1 gene into ${\Delta}Mojmj1$ restored defects in pre-penetration developments including appressorium formation, indicating the importance of histone demethylation through MoJMJ1 during infection-specific morphogenesis. However, defects in penetration and invasive growth were not complemented. We discuss such incomplete complementation in detail here. Our work on MoJMJ1 provides insights into H3K4me3-mediated regulation of infection-specific development in the plant pathogenic fungus.

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1609-1621
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• 2014

The plant pathogen Burkholderia gladioli, which has a broad host range that includes rice and onion, causes bacterial panicle blight and sheath rot. Based on the complete genome sequence of B. gladioli BSR3 isolated from infected rice sheaths, the genome of B. gladioli BSR3 contains the luxI/luxR family of genes. Members of this family encode N-acyl-homoserine lactone (AHL) quorum sensing (QS) signal synthase and the LuxR-family AHL signal receptor, which are similar to B. glumae BGR1. In B. glumae, QS has been shown to play pivotal roles in many bacterial behaviors. In this study, we compared the QS-dependent gene expression between B. gladioli BSR3 and a QS-defective B. gladioli BSR3 mutant in two different culture states (10 and 24 h after incubation, corresponding to an exponential phase and a stationary phase) using RNA sequencing (RNA-seq). RNA-seq analyses including gene ontology and pathway enrichment revealed that the B. gladioli BSR3 QS system regulates genes related to motility, toxin production, and oxalogenesis, which were previously reported in B. glumae. Moreover, the uncharacterized polyketide biosynthesis is activated by QS, which was not detected in B. glumae. Thus, we observed not only common QS-dependent genes between B. glumae BGR1 and B. gladioli BSR3, but also unique QS-dependent genes in B. gladioli BSR3.

• BMB Reports
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• v.57 no.2
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• pp.79-85
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• 2024

Pre-harvest sprouting is a critical phenomenon involving germination of seeds in the mother plant before harvest under relative humid conditions and reduced dormancy. In this paper, we generated HDR mutant lines with one region SNP (C/T) and an insertion of 6 bp (GGT/GGTGGCGGC) in OsERF1 genes for pre-harvest sprouting (PHS) resistance using CRISPR/Cas9 and a geminiviral replicon system. The incidence of HDR was 2.6% in transformed calli. T₁ seeds were harvested from 12 HDR-induced calli and named ERF1-hdr line. Molecular stability, key agronomic properties, physiological properties, and biochemical properties of target genes in the ERF1-hdr line were investigated for three years. The ERF1-hdr line showed significantly enhanced seed dormancy and pre-harvest sprouting resistance. qRT-PCR analysis suggested that enhanced ABA signaling resulted in a stronger phenotype of PHS resistance. These results indicate that efficient HDR can be achieved through SNP/InDel replacement using a single and modular configuration applicable to different rice targets and other crops. This work demonstrates the potential to replace all genes with elite alleles within one generation and greatly expands our ability to improve agriculturally important traits.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.1
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• pp.1-6
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• 1994

Some physicochemical properties of sugary rices were comparatively analyzed with the original cultivars, using Hwachung su-1, su-2, and Nampung su lines bred in Seoul Nat'l Univ., and EM 5 bred in Kyushu Univ. Sugary rice grains were smaller in width and thickness, and the relative dry weight of 1000 grains of sugary rices ranged from 49% to 72%, compared to normal ones. No distinct starch granules were found in endosperm of sugary rices exposured by SEM, with structural variations between Hwachung su-1 and su-2 lines. Sugary rices showed the significant increases in sucrose, glucose and fructose content, especially much higher in sucrose content showing 5.84-8.79%, in comparison with the normal ones, 1.11∼2.65%. However, sucrose and total sugar content varied within sugary lines carrying the same sugary gene, and were significantly lower in Hwachung su-2. Sugary rices, except Hwachung su-1, showed the same or lower ADV and harder gel characteristics, compared to normal ones. Protein and lipid content were much higher in sugary rices, due to the increased relative proportion of aleurone layer in brown rice caused by thicker aleurone layer than in normal rices and the reduction of endosperm part. However, there were no remarkable changes in amino acid and fatty acid compositions.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1467-1475
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• 2015

The use of microbial extracts containing plant hormones is a promising technique to improve crop growth. Little is known about the effect of bacterial cell-free extracts on plant growth promotion. This study, based on phytohormonal analyses, aimed at exploring the potential mechanisms by which Enterococcus faecium LKE12 enhances plant growth in oriental melon. A bacterial strain, LKE12, was isolated from soil, and further identified as E. faecium by 16S rDNA sequencing and phylogenetic analysis. The plant growth-promoting ability of an LKE12 bacterial culture was tested in a gibberellin (GA)-deficient rice dwarf mutant (waito-C) and a normal GA biosynthesis rice cultivar (Hwayongbyeo). E. faecium LKE12 significantly improved the length and biomass of rice shoots in both normal and dwarf cultivars through the secretion of an array of gibberellins (GA₁, GA₃, GA₇, GA₈, GA₉, GA₁₂, GA₁₉, GA₂₀, GA₂₄, and GA₅₃), as well as indole-3-acetic acid (IAA). To the best of our knowledge, this is the first study indicating that E. faecium can produce GAs. Increases in shoot and root lengths, plant fresh weight, and chlorophyll content promoted by E. faecium LKE12 and its cell-free extract inoculated in oriental melon plants revealed a favorable interaction of E. faecium LKE12 with plants. Higher plant growth rates and nutrient contents of magnesium, calcium, sodium, iron, manganese, silicon, zinc, and nitrogen were found in cell-free extract-treated plants than in control plants. The results of the current study suggest that E. faecium LKE12 promotes plant growth by producing GAs and IAA; interestingly, the exogenous application of its cell-free culture extract can be a potential strategy to accelerate plant growth.

• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.186-195
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• 2015

Anthranilate synthase (AS) is a key enzyme in the biosynthesis of tryptophan (Trp), which is the precursor of bioactive metabolites like indole-3-acetic acid and other indole alkaloids. Alpha anthranilate synthase 2 (OsASA2) plays a critical role in the feedback inhibition of tryptophan biosynthesis. In this study, two vectors with single (F124V) and double (S126F/L530D) point mutations of the OsASA2 gene for feedback-insensitive ${\alpha}$ subunit of rice anthranilate synthase were constructed and transformed into wildtype Dongjinbyeo by Agrobacterium-mediated transformation. Transgenic single and double mutant lines were selected as a single copy using TaqMan PCR utilized nos gene probe. To select intergenic lines, the flanking sequence of RB or LB was digested with a BfaI enzyme. Four intergenic lines were selected using a flanking sequence tagged (FST) analysis. Expression in rice (Oryza sativa L.) of the transgenes resulted in the accumulation of tryptophan (Trp), indole-3-acetonitrile (IAN), and indole-3-acetic acid (IAA) in leaves and tryptophan content as a free amino acid in seeds also increased up to 30 times relative to the wildtype. Two homozygous event lines, S-TG1 and D-TG1, were selected for characterization of agronomic traits and metabolite profiling of seeds. Differentially expressed genes (DEGs), related to ion transfer and nutrient supply, were upregulated and DEGs related to co-enzymes that work as functional genes were down regulated. These results suggest that two homozygous event lines may prove effective for the breeding of crops with an increased level of free tryptophan content.

• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.19-25
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• 2003

Radiation technique in agriculture was initiated to develop mutant rice. Seeds of Daechungbyeo rice were irradiated with 250 Gy gamma ray for the purpose of inducing and selecting rice variants. Some quantitative traits of the variants in M$_{8}$ generation were evaluated and RAPD analysis was carried out. Variants showed a wider range of agronomic characteristics in both a positive and a negative direction compared with their original variety. The new mutants were characterized by an increased or decreased in plant height, lodging resistance and shorter panicle. RAPD analysis showed that polymorphic bands were presented in most of the primers. In comparison with the original variety, variants were classified into four groups through UPGMA analysis. Among mutants no. 91, 139, 140 and 141 was ranked as salt tolerance and the proline content of these mutants was more increased than that of original variety. The lines of 139, 140 and 141 had the highest genetic distance as compared to original variety in the dendrogram. It is expected that such variants will be useful not only for studying molecular genetics but also for breeding research and genetic analysis.s.

• Korean Journal of Breeding Science
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• v.43 no.6
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• pp.532-537
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• 2011

'Seolgaeng' is a new Japonica rice variety developed from an 'Ilpumbyeo' mutant line, N-methyl-N-nitrosourea (MNU) treatment on fertilized egg cell, by a rice breeding team of National Crop Experiment Station, Rural Development Administration in 2001. This variety has about 134 days of growth duration from transplanting to harvesting in central plain area of Korea, a good semi-elect plant type and resistant to lodging with strong culm. The number of panicles/hill of 'Seolgaeng' is more than that of 'Hwaseongbyeo'. The milled rice appearance of this cultivar is an opaque, but it has amylose content (19.3%) similar to 'Hwaseongbyeo'. It is superior to hypae formation of Aspergillus orzyae in the making of fermented rice and amount of pigmentation in fermented rice by Monascus anka that of 'Ilpumbyeo'. This variety showed slow leaf senescence and considerable resistance to viviparous germination. It is moderately resistant to leaf blast and susceptible to bacterial blight, virus disease and insect pests. The yield performance of this rice cultivar is about 5.27 MT/ha in milled rice in local adaptability test for three years from 1999 to 2001). 'Seolgaeng' is adaptable to central and southern plain areas of Korea.

• Journal of Photoscience
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• v.9 no.2
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• pp.90-93
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• 2002

Phytochrome A (phyA) and phytochrome B (phyB) perceive light and adapt to fluctuating circumstances by different manners in terms of effective wavelengths, required fluence and photoreversibility. Action spectra for induction of seed germination and inhibition of hypocotyl elongation using phytochrome mutants of Arabidopsis showed major difference. PhyA is the principal photoreceptor for the very low fluence responses and the far-red light-induced high irradiance responses, while phyB controls low fluence response in a red/far-red reversible mode. The structural requirement of their bilin chromophores for photosensory specificity of phyA and phyB was investigated by reconstituting holophytochromes through feeding various synthetic bilins to the following chromophore-deficient mutants: hy1, hyl/phyA and hyl/phyB mutants of Arabidopsis. We found that the vinyl side-chain of the D-ring in phytochromobilin interacts with phyA apoprotein. This interaction plays a direct role in mediating the specific photosensory function of phyA. The ethyl side-chain of the D-ring in phycocyanobilin fails to interact with phyA apoprotein, therefore, phyA specific photosensory function is not observed. In contrast, both phytochromobilin and phycocyanobilin interact with phyB apoprotein and induce phyB specific photosensory functions. Structural requirements of the apoproteins and the chromophores for the specific photoperception of phyA and phyB are discussed.

• The Plant Pathology Journal
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• v.20 no.3
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• pp.229-233
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• 2004

The bacterial leaf blight, which is caused by Xantho-monas oryzae pv. oryzae, is the most damaging and intractable disease of rice. To identify the genes involved in the virulence mechanism of transposon TnS complex, which possesses a linearized transposon and transposase, was successfully introduced into X. oryzae pv. oryzae by electroporation. The transposon mutants were selected and confirm the presence of transposition in X. oryzae pv. oryzae by the PCR amplification of transposon fragments and the Southern hybridization using these mutants. Furthermore, transposon insertion sites in the mutant bacterial chromosome were deter-mined by direct genomic DNA sequencing using transposon-specific primers with ABI 3100 Genetic Analyzer. Efficiency of transposition was influenced mostly by the competence status of X. oryzae pv. oryzae cells and the conditions of electroporation. These results indicated that the insertion mutagenesis strategy could be applied to define function of uncharacterized genes in X. oryzae pv. oryzae.