• The Plant Pathology Journal
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• v.26 no.4
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• pp.313-320
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• 2010

Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.

• The Plant Pathology Journal
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• v.35 no.6
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• pp.564-574
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• 2019

Like many fungal pathogens, the conidium and appressorium play key roles during polycyclic dissemination and infection of Magnaporthe oryzae. Ran-binding protein microtubule-organizing center (RanBPM) is a highly conserved nucleocytoplasmic protein. In animalia, RanBPM has been implicated in apoptosis, cell morphology, and transcription. However, the functional roles of RanBPM, encoded by MGG_00753 (named MoRBP9) in M. oryzae, have not been elucidated. Here, the deletion mutant ΔMorbp9 for MoRBP9 was generated via homologous recombination to investigate the functions of this gene. The ΔMorbp9 exhibited normal conidial germination and vegetative growth but dramatically reduced conidiation compared with the wild type, suggesting that MoRBP9 is involved in conidial production. ΔMorbp9 conidia failed to produce appressoria on hydrophobic surfaces, whereas ΔMorbp9 still developed aberrantly shaped appressorium-like structures at hyphal tips on the same surface, suggesting that MoRBP9 is involved in the morphology of appressorium-like structures from hyphal tips and is critical for development of appressorium from germ tubes. Taken together, our results indicated that MoRBP9 played a pleiotropic role in polycyclic dissemination and infection-related morphogenesis of M. oryzae.

• Korean journal of applied entomology
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• v.13 no.4 s.21
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• pp.245-249
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• 1974

In order to establish a dose limits for subsequent induced mutation research itl Pyricularia oryzae, X-ray sensitivity of the conidia and the vegetative hypae of the fungus, race N-1, was investigated. Conidia of the fungus irradiated with X-rays reduced significantly in spore germination inversely with radiation doses. A severe suppression of conidia germination in about $80\%$ was found at the dose of 120kR, and the rests of the conidia produce very short and lysed germ tubes. A stimulated effect was observed in the elongation of hyphae from the conidia of 10 kR irradiation at initial stage of the growth. The radiosensitivity of hyphae was exremely higher than that of conidia with the increase of radiation doses. It was also recognized that the frequency of X-ray induced mutation in pathogenicity was directly proportional to radiation doses.

• The Korean Journal of Mycology
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• v.17 no.1
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• pp.1-6
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• 1989

The mycelia of Pyricularia oryzae were treated with enzyme solution mixture consisting of Driselase, ${\beta}-Glucuronidase$, Cellulase, and Macerozyme R-10 for the production of protoplasts. More protoplasts were formed from mycelia of race KJ 101 of P. oryzae than that of race KI 315a in the enzyme mixtures. The number of protoplasts was decreased in the untreated control three hrs after the enzyme treatment, whereas the number was increased in the treatments with 10, 50 and 100 mM 2-mercaptoethanol, respectively. The number of protoplasts increased to reach maximum at five hrs after treatment of 10 mM 2-mercaptoethanol, but the least was found in 200 mM. The protoplasts of P. oryzae in a liquid medium containing 2.5% yeast extract, and 2% dextrose reverted to the mycelia after five hrs shaking incubation at $27^{\circ}C$. Some protoplasts produced yeast like buds and the buds were developed to irregularly shaped chains of cell protruded a germ tube like hypha from the distal cell. Once in a while a germ tube like hypha protruded directly from the protoplasts. Except in the first type of reversion, other protoplasts reverted to the normal mycelia. The reversion frequency was highest on PDA with stabilizer of 0.6 M KCl. No reversion of protoplasts occurred on water agar regaardless oftreaatments.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.27 no.4
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• pp.371-384
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• 1982

Highly cold tolerant varieties are requested not only at high latitute cool area but also tropical high elevated areas, and the required tolerance is different from location to location. IRRI identified 6 different types of cold tolerance required in the world for breeding purpose; a) Hokkaido type, b) Suweon type, c) Taipei 1st season type, d) Taipei 2nd season type, e) Tropical alpine type and, f) Bangladesh type. The cold tolerance requested in Korea is more eargent in Tongil group cultivars and their required tolerance is the one such as the physiological activities at low temperature are as active as in Japonica group cultivars at least during young seedling stage and reproduction stage. With conventional Japonica cultivars, such cold tolerant characters are requested as short growth duration but stable basic vegetative growth, less sensitive to high temperature and less prolonged growth duration at low temperature. The methods screening for cold tolerance were developed rapidly after the Tongil cultivar was reliesed. The facilities of screening for cold tolerance, such as, low temperature incubator, cold water tank, growth cabinet, phytotron, cold water nursery in Chuncheon, breeding nursery located in Jinbu, Unbong and Youngduk, are well established. Foreign facilities such as, cold water tank with the rapid generation advancement facilities, cold nurseries located in Banaue, Kathmandu and Kashimir may be available for the screening of some limitted breeding materials. For the reference, screening methods applied at different growth stages in Japan are introduced. The component characters of cold tolerance are not well identified, but the varietal differences in a) germinability, b) young seedling growth, c) rooting, d) tillering, e) discolation, f) nutrition uptake, g) photosynthesis rate, h) delay in heading, i) pollen sterility, and j) grain fertility at low temperature are reported to be distinguishable. Relationships among those traits are not consistent. Reported studies on the inheritance of cold tolerance are summarized. Four or more genes are controlling low temperature germinability, one or several genes are controlling seedling tolerance, and four or more genes are responsible for the pollen fertility of the rice treated with cold air or grown in the cold water nursery. But most of those data indicate that the results may come out in different way if those were tested at different temperature. Many cold tolerant parents among Japonicas, Indicas and Javanicas were identified as the results of the improvement of cold tolerance screening techniques and IRTP efforts and they are ready to be utilized. Considering a) diversification of germ plasm, b) integration of resistances to diseases and insects, c) identification of adaptability of recommending cultivars and, d) systematic control of recommending cultivars, breeding strategies for short term and long term are suggested. For short term, efforts will be concentrated mainly to the conventional cultivar group. Domestic cultivars will be used as foundation stock and ecologically different foreign introductions such as from Hokkaido, China or from Taiwan, will be used as cross parents for the adjustment of growth durations and synthsize the prototype of tolerances. While at the other side, extreme early waxy Japonicas will be crossed with the Indica parents which are identified for their resistances to the diseases and insects. Through the back corsses to waxy Japonicas, those Indica resistances will be transfered to the Japonicas and these will be utilized to the crosses for the improvement of resistances of prototype. For the long term, efforts will be payed to synthsize all the available tolerances identified any from Japonicas, Indicas and Javanicas to diversify the germ plasm. The tolerant cultivars newly synthsized, should be stable and affected minimum. to the low temperature at all the growing stages. The resistances to the diseases and insects should be integrated also. The rapid generation advancement, pollen culture and international cooperations were emphasized to maximize the breeding efficiency.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.10
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• pp.1454-1464
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• 2015

Four methods were adopted, including the mobile nylon bag (MNB) method, modified three-step in vitro (MTS) method, original three-step in vitro (OTS) method, and acid detergent insoluble nitrogen (ADIN) estimating method, to evaluate the intestinal digestibility of rumen undegradable protein (DRUP) of 10 types of concentrates and 7 types of roughages. After correlation analysis to determine the DRUP values using the MNB, MTS, OTS, and ADIN methods, the study aimed to find out appropriate methods to replace the MNB method due to its disadvantages such as high price, long time period, and use of a duodenal T-fistula. Three dairy cows with a permanent ruminal fistula and duodenal T-fistula were used in a single-factor experimental design. The results showed that the determined DRUP values using the MNB method for soybean meal, cottonseed meal, rapeseed meal, sunflower meal, corn germ meal, corn, rice bran, barley, wheat bran, corn fiber feed, Alfalfa (Zhao dong), Alfalfa (Long mu 801), Alfalfa (Long mu 803), grass (North), Grass (Inner Mongolia), corn silage and corn straw were 98.13%, 87.37%, 88.47%, 82.60%, 75.40%, 93.23%, 69.27%, 91.27%, 72.37%, 79.03%, 66.72%, 68.64%, 73.57%, 50.47%, 51.52%, 54.05%, and 43.84%, respectively. The coefficient of determination ($R^2=0.964$) of the results between the MTS method and the MNB method was higher than that ($R^2=0.942$) between the OTS method and the MNB method. The coefficient of determination of the DRUP values of the concentrates among the in vitro method (including the MTS and OTS methods) and the MNB method was higher than that of the roughage. There was a weak correlation between the determined DRUP values in concentrates obtained from the ADIN method and those from the MNB method, and there was a significant correlation (p<0.01) between the determined DRUP values of the roughage obtained from the MNB method and those obtained from ADIN method. The DRUP values were significantly correlated with the nutritional ingredients of the feeds. The regression equation was DRUP =100.5566+0.4169CP - 0.4344SP - 0.7102NDF - 0.7950EE ($R^2=0.8668$, p<0.01; CP, crude protein; SP, soluble protein; NDF, neutral detergent fiber; EE, ether extract). It was concluded that both the MTS method and the OTS may suitable to replace the MNB method for determining the DRUP values and the former method was more effective. Only the ADIN method could be used to predict the values of the roughages but conventional nutritional ingredients were available for all of the samples' DRUP.