Rice blast caused by Pyricularia oryzae, which is a major threat to food security worldwide, markedly decreases the yield of rice. Some rhizobacteria called 'plant growth-promoting rhizobacteria' inhibit plant pathogens and improve plant growth by secreting iron-chelating siderophores. The decreased availability of iron adversely affects the survival of pathogens, especially fungal pathogens, in the rhizosphere. This study aimed to determine the morphological diversity of siderophore-producing bacteria, analyze the type of siderophores produced by the bacteria, and examine their growth-inhibitory activity against Pyricularia oryzae. The rhizobacteria were isolated from the rhizosphere of Sembada Hitam variety of black rice plants in Pakem, Sleman, Yogyakarta, Indonesia. In total, 12 distinct isolates were screened for the production of siderophores. It was found that 9 out of 12 bacteria produced siderophore and most of them were Gram positive bacteria. The best siderophore-producing isolates with different type of siderophore were used in further studies. The IS3 and IS14 isolates were found to be the best siderophore producer that produced hydroxamate and mixed type of hydroxamate-carboxylate type of siderophore, respectively. In the dual culture assay, IS14 showed a strong antagonistic effect against Pyricularia oryzae by the 81.17% inhibition.
Planthopper infestation in rice causes direct and indirect damage through feeding and viral transmission. Host microbes and small RNAs (sRNAs) play essential roles in regulating biological processes, such as metabolism, development, immunity, and stress responses in eukaryotic organisms, including plants and insects. Recently, advanced metagenomic approaches have facilitated investigations on microbial diversity and its function in insects and plants, highlighting the significance of microbiota in sustaining host life and regulating their interactions with the environment. Recent research has also suggested significant roles for sRNA-regulated genes during rice-planthopper interactions. The response and behavior of the rice plant to planthopper feeding are determined by changes in the host transcriptome, which might be regulated by sRNAs. In addition, the roles of microbial symbionts and sRNAs in the host response to viral infection are complex and involve defense-related changes in the host transcriptomic profile. This review reviews the structure and potential functions of microbes and sRNAs in rice and the associated planthopper species. In addition, the involvement of the microbiota and sRNAs in the rice-planthopper-virus interactions during planthopper infestation and viral infection are discussed.
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.134-134
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2017
Recently, host-induced gene silencing (HIGS) system has been successfully applied into development of resistant crops against insects, fungal and viral pathogens. To test HIGS-mediated resistance in rice against rice blast fungus, Magnaporthe oryzae, we first tested possibility of movement of small non-coding RNA from rice cells to rice blast fungus. The rice blast fungus expressing GFP transgene were inoculated to transgenic rice plants ectopically expressing dsRNAi construct targeting fungal GFP gene. Expression of dsRNAi construct for GFP gene in transgenic plants significantly suppressed GFP expression in infected fungal cells indicating that small RNAs generated in plant cells can move into infected fungal cells and efficiently suppress the expression of fungal GFP gene. Consistent with these results, expression of dsRNAi constructs against 3 fungal pathogenic genes of M. oryzae in transgenic rice specifically and efficiently suppressed not only the expression of fungal pathogenic genes, but also fungal infection. The conidia of M. oryzae applied on leaf sheath of transgenic rice expressing dsRNAs against 3 fungal pathogenic genes showed abnormal development of primary hyphae and malfunction of appressorium, which is consistent with the phenotypes of corresponding fungal knock-out mutants. Taken these results together, here, we suggest a novel strategy for development of antifungal crops by means of HIGS system.
Koo, Sung Cheol;Choi, Man Soo;Chun, Hyun Jin;Shin, Dong Bum;Park, Bong Soo;Kim, Yul Ho;Park, Hyang-Mi;Seo, Hak Soo;Song, Jong Tae;Kang, Kyu Young;Yun, Dae-Jin;Chung, Woo Sik;Cho, Moo Je;Kim, Min Chul
Molecules and Cells
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v.27
no.5
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pp.563-570
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2009
We previously isolated the OsCBT gene, which encodes a calmodulin (CaM)-binding protein, from a rice expression library constructed from fungal elicitor-treated rice suspension cells. In order to understand the function of OsCBT in rice, we isolated and characterized a T-DNA insertion mutant allele named oscbt-1. The oscbt-1 mutant exhibits reduced levels of OsCBT transcripts and no significant morphological changes compared to wild-type plant although the growth of the mutant is stunted. However, oscbt-1 mutants showed significant resistance to two major rice pathogens. The growth of the rice blast fungus Magnaporthe grisea, as well as the bacterial pathogen Xanthomonas oryzae pv. oryzae was significantly suppressed in oscbt-1 plants. Histochemical analysis indicated that the hypersensitive-response was induced in the oscbt-1 mutant in response to compatible strains of fungal pathogens. OsCBT expression was induced upon challenge with fungal elicitor. We also observed significant increase in the level of pathogenesis-related genes in the oscbt-1 mutant even under pathogen-free condition. Taken together, the results support an idea that OsCBT might act as a negative regulator on plant defense.
Rice yield and protein content have been shown to be highly variable across paddy fields. In order to characterize this spatial variability of rice within a field, two-year experiments were conducted in 2002 and 2003 in a large-scale rice field of $6,600m^2$ In year 2004, an experiment was conducted to know if variable rate treatment (VRT) of N fertilizer, that was prescribed for site-specific management at panicle initiation stage, could reduce spatial variation in yield and protein content of rice while increasing yield compared to conventional uniform N topdressing (UN, 33kg N/ha at PIS) method. VRT nitrogen prescription for each grid was calculated based on the nitrogen (N) uptake (from panicle initiation to harvest) required for target rice protein content of $6.8\%$, natural soil N supply, and recovery of top-dressed N fertilizer. The required N uptake for target rice protein content was calculated from the equations to predict rice yield and protein content from plant growth parameters at panicle initiation stage (PIS) and N uptake from PIS to harvest. This model· equations were developed from the data obtained from the previous two-year experiments. The plant growth parameters for the calculation of the required N were predicted non-destructively by canopy reflectance measurement. Soil N supply for each grid was obtained from the experiment of year 2003, and N recovery was assumed to be $60\%$ according to the previous reports. The prescribed VRT N ranged from 0 to 110kg N/ha with an average of 57kg/ha that was higher than 33 kg/ha of UN. The results showed that VRT application successfully worked not only to reduce spatial variability of rice yield and protein content but also to increase rough rice yield by 960kg/ha. The coefficient of variation (CV) for rice yield and protein content was reduced significantly to $8.1\%$ and $7.1\%$ in VRT from $14.6\%$ and $13.0\%$ in UN, respectively. And also the average protein content of milled rice in VRT showed very similar value of target protein content of $6.8\%$. In conclusion the procedure used in this paper was believed to be reliable and promising method for reducing within-field spatial variability of rice yield and protein content. However, inexpensive, reliable, and fast estimation methods of natural N supply and plant growth and nutrition status should be prepared before this method could be practically used for site-specific crop management in large-scale rice field.
Salt injury in rice is caused mainly by the salinity in soil and in the irrigated water, and occasionaly by salinity delivered through typhoon from the sea. The salt concentration of rice plants increased with higher salinity in the soil of the rice growing. The climatic conditions, high temperature and solar radiation and dry conditions promote the salt absorption of rice plant in saline soil. The higher salt accumulation in the rice plant generally reduces the root activity and inhibits the absorption of minerals of rice plant, resulting the reduction of photosynthesis. The salt damages of rice plant, however, are different from different growth stage of rice plants as follows: 1. Germination of rice seed was slightly delayed up to 1.0% of salt concentration and remarkably at 1. 5%, but none of rice seeds were germinated at 2.5%. This may be due to the delayed water uptake of rice seeds and the inhibition of enzyme activity, 2. It was enable to establish rice seedlings at seed bed by 0.2% of salt concentration with some reduction of leaf elongation. The increasing of 0.3% salt concentration caused to the seedling death with varietal differences, but most of seedlings were death at 0.4% with no varietal differences. 3. Seedlings grown at the nursery over 0.1% salt, gradually reduced in rooting activity after transplanting according to increasing the salt concentration from 0.1% up to 0.3% of paddy field. However, the seedlings grown in normal seed bed showed no difference in rooting between varieties up to 0.1% but significantly different at 0.3% between varieties, but greatly reduced at 0.5% and died at last in paddy after transplanting. 4. At panicle initiation stage, rice plant delayed in heading by salt damage, at meiotic stage reduced in grains and its filling rate due to inhibition of glume and pollen developing, and salt damage at heading stage and till 3 weeks after heading caused to reduction of fertilization and ripening rate. In viewpoint of agricultural policy the overcoming strategy for salt injury is to secure sufficient water source. Irrigation and drainage systems as well as underground drainage is necessary to desalinize more effectively. This must be the most effective and positive way except cost. By cultural practice, growing the salt tolerant variety with high population could increase yield. The intermittent irrigation and fresh water flooding especially at transplanting and from panicle initiation to heading stage, the most sensitive to salt injury, is important to reduce the salt content in saline soil. During the off-cropping season, plough and rotavation with flooding followed by drainage, or submersion and drainage with groove could improve the desalinization. Increase of nitrogen fertilizer with more split application, and soil improvement by lime, organic matter and forign soil addition, could increase the rice yield. Shift of trans-planting is one of the way to escape from the salt injury.
Methanol extracts of fresh materials of 57 plants were screened for in vivo antifungal activity against Magna-porthe grisea, Corticium sasaki, Botrytis cinerea, Phyto-phthora infestans, Puccinia recondita, and Blumeria graminis f. sp. hordei. Among them, seven plant extracts showed disease-control efficacy of more than 90% against at least one of six plant diseases. None of the plant extracts was highly active against tomato gray mold. The methanol extracts of Chloranthus japonicus (roots) (CjR) and Paulownia coreana (stems) (PcS) displayed the highest antifungal activity; the CjR extract controlled the development of rice blast, rice sheath blight, and wheat leaf rust more than 90%, and tomato gray mold and tomato late blight more than 80%. The PcS extract displayed control values of more than 90 % against rice blast, wheat leaf rust, and barley powdery mildew and more than 80% against tomato gray mold. The extract of PcS also had a curative activity against rice sheath blight and that of CjR had a little curative activity against rice blast. On the other hand, the extract of Rumex acetocella roots reduced specifically the development of barley powdery mildew. Further studies on the characterization of antifungal substances in antifungal plant extracts are underway and their disease-control efficacy should be examined under greenhouse and field conditions.
This study was carried out to obtain the basic information for breeding cold-tolerant rice varieties with high yield-productivity through wide crosses between indica and japonica rice. Genetic analysis was conducted using 55 F$_1$s obtained from half-diallel crosses among eleven cultivars of various origin including indica and japonica rice. Screening for cold tolerance was done with cold-water irrigation after transplanting until ripening stage. Both general combining ability (GCA) and specific combining ability (SCA) effects were highly significant in all characters associated with dry matter accumulation at 30 and 50days after cold-water irrigation (DAC). The variance of GCA was much larger than that of SCA in plant height, shoot dry weight per plant (DWP), crop growth rate (CGR) and cold-water response index (CRI) of these characters except CRI of shoot dry weight per plant. The DWP, CGR and CRI of these characters of Gaochan 102, Tong88-7 and TR22183 were markedly higher than those of the others. GCA effects of these varieties on DWP, CGR and their CRI were also higher than those of the others, indicating that they are useful as promising parents for breeding cold-tolerant varieties. Analysis of genetic parameters for 11$\times$11 half-diallel F$_1$s revealed that inter-locus gene interaction were concerned in the expression of plant height at 50 DAC, CRI of DWP at 50 DAC, and CRI of CGR, and that intra-locus gene interaction for plant height and the other characters were partial dominance and over-dominance, respectively. Narrow-sense heritability (h$^2$$_{N}$) was the highest in plant height as 0.729, and the lowest in CRI of DWP at 30 DAC as 0.048, suggesting that selection for cold tolerance will be quite effective in case that the selection criterion is the performance itself.f.
To identify a novel biopesticide controlling rice blast disease caused by Magnaporthe oryzae, 700 plant extracts were evaluated for their inhibitory effects on mycelial growth of M. oryzae. The L. foenum-graecum Herba extract showed the lowest inhibition concentration ($IC_{50}$) of $39.28{\mu}g/ml$, which is lower than the $IC_{50}$ of blasticidin S ($63.06{\mu}g/ml$), a conventional fungicide for rice blast disease. When treatments were combined, the $IC_{50}$ of blasticidin S was dramatically reduced to $10.67{\mu}g/ml$. Since ABC transporter genes are involved in fungicide resistance of many organisms, we performed RT-PCR to investigate the transcriptional changes of 40 ABC transporter family genes of M. oryzae treated with the plant extract, blasticidin S, and tetrandrine, a recognized ABC transporter inhibitor. Four ABC transporter genes were prominently activated by blasticidin S treatment, but were suppressed by combinational treatment of blasticidin S with the plant extract, or with tetrandrine that didn't show cellular toxicity by itself in this study. Mycelial death was detected via confocal microscopy at 24 h after plant extract treatment. Finally, subsequent rice field study revealed that the plant extract had high control efficacy of 63.3% and should be considered a biopesticide for rice blast disease. These results showed that extract of L. foenum graecum Herba suppresses M. oryzae ABC transporter genes inducing mycelial death and therefore may be a potent novel biopesticide.
Wu, Jingni;Kim, Sang Gon;Kang, Kyu Young;Kim, Ju-Gon;Park, Sang-Ryeol;Gupta, Ravi;Kim, Yong Hwan;Wang, Yiming;Kim, Sun Tae
The Plant Pathology Journal
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v.32
no.6
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pp.552-562
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2016
Pathogenesis-related proteins play multiple roles in plant development and biotic and abiotic stress tolerance. Here, we characterize a rice defense related gene named "jasmonic acid inducible pathogenesis-related class 10" (JIOsPR10) to gain an insight into its functional properties. Semi-quantitative RT-PCR analysis showed up-regulation of JIOsPR10 under salt and drought stress conditions. Constitutive over-expression JIOsPR10 in rice promoted shoot and root development in transgenic plants, however, their productivity was unaltered. Further experiments exhibited that the transgenic plants showed reduced susceptibility to rice blast fungus, and enhanced salt and drought stress tolerance as compared to the wild type. A comparative proteomic profiling of wild type and transgenic plants showed that overexpression of JIOsPR10 led to the differential modulation of several proteins mainly related with oxidative stresses, carbohydrate metabolism, and plant defense. Taken together, our findings suggest that JIOsPR10 plays important roles in biotic and abiotic stresses tolerance probably by activation of stress related proteins.
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