Null mutants generated by targeted gene replacement are frequently used to reveal function of the genes in fungi. However, targeted gene deletions may be difficult to obtain or it may not be applicable, such as in the case of redundant or lethal genes. Constitutive expression system could be an alternative to avoid these difficulties and to provide new platform in fungal functional genomics research. Here we developed a novel platform for functional analysis genes in Magnaporthe oryzae by constitutive expression under a strong promoter. Employing a binary vector (pGOF1), carrying $EF1{\beta}$ promoter, we generated a total of 4,432 transformants by Agrobacterium tumefaciens-mediated transformation. We have analyzed a subset of 54 transformants that have the vector inserted in the promoter region of individual genes, at distances ranging from 44 to 1,479 bp. These transformants showed increased transcript levels of the genes that are found immediately adjacent to the vector, compared to those of wild type. Ten transformants showed higher levels of expression relative to the wild type not only in mycelial stage but also during infection-related development. Two transformants that T-DNA was inserted in the promotor regions of putative lethal genes, MoRPT4 and MoDBP5, showed decreased conidiation and pathogenicity, respectively. We also characterized two transformants that T-DNA was inserted in functionally redundant genes encoding alpha-glucosidase and alpha-mannosidase. These transformants also showed decreased mycelial growth and pathogenicity, implying successful application of this platform in functional analysis of the genes. Our data also demonstrated that comparative phenotypic analysis under over-expression and suppression of gene expression could prove a highly efficient system for functional analysis of the genes. Our over-expressed transformants library would be a valuable resource for functional characterization of the redundant or lethal genes in M. oryzae and this system may be applicable in other fungi.
Three species of Fusarium, F. fujikuroi, F. verticillioides and F. proliferatum, are known to be associated with bakanae disease of rice [1, 2]. F. fujikuroi infects rice flowers and survive in endosperm and embryo of the seeds. Infected seed is an important source of primary inoculum of pathogens [3]. Seeds of rice (Oryza sativa cv. Boramchan) collected from bakanae-infected field were found to be 96% infected with Fusarium sp., 52% with F. fujikuroi, 42% with F. verticillioides, and 12% with F. proliferatum as determined by incubation method and species-specific PCR assays. F. fujikuroi was detected at lemma/palea, endosperm and embryo whereas F. verticillioides and F. proliferatum were recovered only from lemma/palea by means of component plating test. Seed disinfection methods have been developed to control bakanae disease and prochloraz has been most widely used for rice seeds. Two chemicals formulated with prochloraz (PC 1) and prochloraz + hexaconazole (PC 2) that inhibit biosynthesis of ergosterol strongly reduced the incidence of Fusarium spp. on selective media to 4.7% and 2.0%, respectively. Disease symptoms of rice seedlings in nursery soil were alleviated by chemical treatment; seedlings with elongated leaves or wide angle between leaf and stem were strikingly reduced from 15.6 to 3.2% (PC 1) and 0 (PC 2), stem rots were reduced from 56.9 to 26.2% (PC 1) and 32.1% (PC 2), and normal seedling increased from 0.4 to 13.3% (PC 2). Prochloraz has some disadvantages and risks such as the occurrence of tolerant pathogens [4] and effects on the sterol synthesis in animals and humans [5]. For these reasons, it is necessary to develop new disinfection method that do not induce fungal tolerance and are safe to humans and animals. Chlorine dioxide ($ClO_2$), that is less toxic, produces no harmful byproducts, and has high oxidizing power, has been reported to be effective at disinfection of several phytopathogenic fungi including Colletotrichum spp. and Alternaria spp. [6]. Gaseous $ClO_2$ applied to rice seeds at a concentration of 20 ppm strongly suppressed mycelial growth of Fusarium fujikuroi, F. verticillioides and F. proliferatum. The incidence of Fusarium spp. in dry seed with 8.7% seed moisture content (SMC) tended to decrease as the concentration of $ClO_2$ increased from 20 to 40 ppm. Applying 40 ppm $ClO_2$ at 90% relative humidity, incidence was reduced to 5.3% and resulted in significant reduction of disease symptoms on MS media. In nursery soil, stem rot was reduced from 56.9 to 15.4% and the number of normal seedlings increased from 0.4 to 25.5%. With water-soaked seeds (33.1% SMC) holding moisture in the endosperm and embryo, the effectiveness of disinfection using $ClO_2$ increased, even when treated with only 20 ppm for four hours. This suggests that moisture was a key element for action of $ClO_2$. Removal of the palea and lemma from seeds significantly decreased the incidence of Fusarium spp. to 3.0%. Seed germination appeared to decrease slightly by water-soaking at $30^{\circ}C$ because of increased SMC and by physical damage of embryos from hulling. These results indicate that the use of gaseous $ClO_2$ was effective as a means to disinfect rice seeds infected with Fusarium spp. and that moisture around the pathogens in the seed was an important factor for the action of $ClO_2$. Further investigations should be conducted to ascertain the best conditions for complete disinfection of Fusarium spp. that infect deep site of rice seeds.
In order to establish the scientific foundations for the production of Korean traditional kanjang (soy sauce) in a semi-pilot scale, meju preparation and kanjang mashing methods were investigated. Two types of meju, one that was prepared by conventional method (CM-meju) and the other that prepared by modified conventional method (MCM-meju), were made and compared their characteristics. The former made of cooked and crushed soybean with a brick shape was fermented at $15{\sim}20^{\circ}C\;and\;40{\sim}50%$ relative humidity (RH) for 30 days in koji room and the latter was fermented at $25{\sim}30^{\circ}C\;and\;80{\sim}90%$ RH seemingly being optimum for the growth of meju-organisms for the same period. The quality of MCM-meju as a raw material for the kanjang preparation was considered to be superior to that of CM-meju as the higher soluble nitrogen and total free amino acids content, the major factor for the evaluation of the quality of meju, were found in the former although the higher total nitrogen content and lower % weight loss were observed in the latter during meju preparation process. The quality of MCM-kanjang with higher total nitrogen, free amino acids, free sugars and the lower residual nitrogen content in cake after separation of kanjang was also found to be superior to that of CM-kanjang in sensory evaluation results.
Forest soil fertility was evaluated through the approach of numerical method. In this study, the soil chemical properties analyzed for 35 different soil series as table 2 were cited in numerical analysis. Minimum contents of essential nutrients in the surface soil for a satisfactory growth of tree in the plantation were evaluated by comparing with Wild's standard as table 1. Demanding level of fertilization were evaluated by using the formula 1 as table 5. Similar relation of soil chemical properties between soil series were calculated through formula 2, and then classified into 5 groups in soil chemical properties. 1. General chemical properties of surface soil in case of 35 soil series. About 40 percent of 35 different soil series are less than 2 percent in organic matter, 10 ppm in available phosphorus, 1.25m.e/l00g in exchangeable calcium and 0.5m.e/l00g in exchangeable magnesium. Generally, shortage of exchangeable potash are not found. CEC less than 10m.e/l00g are in two thirds and strong acid soil less than PH 5.5 are in about four fifths. 2. Soil series requested or not the fertilization are indirectly evaluated from the formula 1 using the relative figure of chemical components of CEC, OM and MgO. Through this analysis, 8 different soil series have very poor quality in soil chemical capacity so that demands highly the fertilization. On the other hand, other 13 different soil series group have not been thought to need the fertilization according to chemical guality. 3. By the results comparing the similarity of chemical properties of forest soil, it is thought to be suitable that the forest soil fertility are divided into 5 groups as follows: 1. Low CEC soil 1-1 Low organic matter soil less than 2 percent 1-2 Medium organic matter soil less than 4 percent 2. High CEC and organic matter soil 2-1 Low magnesium soil 2-2 High magnesium soil 3. High magnesium and calcium soil as lime stone.
Objectives : Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) is a hair care product which is composed of ten plant extracts used in oriental medicine. This study was carried out to investigate the effects of Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) on hair regrowth and cytokine changes in a shaving model of C57BL/6 mice. Materials and Methods : Five-week-old mice were acclimated for 1 week at a temperature between $21-23^{\circ}C$, 40-60% relative humidity, and 12h of a light/dark cycle before beginning of the experiment. There were three experimental groups including 50% ethanol (EtOH, control), a positive control of 3% Minoxidil, and 30% Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) in 50% ethanol in 18 female mice. The test compounds were topically treated once a day over 12 days. The hair regrowth was photographically and histologically determined during the experimental period of 12 days. Revelation of EGF, $TGF-{\beta}1$ and IL-6 in hair follicle were also determined using immunohistochemistry. In addition to that, IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ in skin tissue were determined using enzyme-linked immunosorbent assay(ELISA). Results : Hair regrowth in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups was promoted earlier and faster than the control group. Concentrations of hairs and thick-hair ratio in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups were promoted than the control group. EGF was moderately positive in hair follicle of 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups, but negative in the control group. $TGF-{\beta}1$ was not significantly difference between the groups. IL-6 in hair follicle of Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) group was negative, but weakly positive in 3% Minoxidil and control group. IL-6 and $IL-1{\beta}$ in skin tissue were significantly decreased in Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) group, but there was not significantly decreased in 3% Minoxidil and control group. $TNF-{\alpha}$ in skin tissue was significantly decreased in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups. Conclusions : These results suggest that Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) has hair growth promoting activity and it can be used for treatment of alopecia. And these effects relate to EGF revelation of hair follicle and a decrease IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ in skin tissue.
Lee, Hee Ju;Lee, Sang Gyu;Kim, Sung Kyeom;Choi, Chang Sun;Kim, Sang Beom
Journal of Bio-Environment Control
/
v.25
no.3
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pp.162-167
/
2016
This study was carried out to investigate proper stem cutting position and air temperature to regenerate roots from cuttings of bitter gourd. 'NS454' (NS) and 'Dragon' (DR) cultivars were tested and the cutting position was cut at the 3rd node in the stem (treatment I) and cut at the central part (stem segment) between 3rd leaf and 4th leaf in the stem (treatment II). The air temperature was maintained at 18, 23, 28 and $35^{\circ}C$ in the growth chambers, respectively. The photosynthetic photon flux at the ground of a chamber was maintained in approximately $150-200{\mu}mol\;m^{-2}s^{-1}$ during the 16-hour photoperiod. The relative humidity in the chambers was maintained over 85%. After 10 days of cuttings, regardless of cutting position, DR and NS cultivars showed 100% the survival rate in the $18^{\circ}C$ or $23^{\circ}C$ treatments, however, most of plants $18^{\circ}C$ treatment had not rooting. In the $28^{\circ}C$ treatment, regardless of cutting position, DR and NS cultivars showed 100% of the rooting rate. In the $23^{\circ}C$ treatment, for DR cultivar, the node cutting showed 90% of rooting rate and the stem segment cutting showed 40% rooting rate. For NS cultivar, the node cutting showed 50% of rooting rate and the stem segment cutting showed 40% rooting rate. DR and NS cultivars have high rooting rate, while the rooting rate remarkably decreased in the $35^{\circ}C$ treatment. For DR cultivar, the stem segment cutting showed 5.3 of the highest rooting number and the node cutting showed 2.7 rooting number in the $28^{\circ}C$ treatment. NS cultivar was not significantly different the rooting regardless of cutting position. The results suggested that cutting at the stem segment cutting of bitter gourd is an effective cutting method for increasing survival rate and the air temperature should be maintained at $28^{\circ}C$.
Fruiting bodies of Cordyceps have been regarded as popular folk and effective medicines to treat human diseases such as asthma, bronchial and lung inflammation, and kidney disease. Cordyceps pruinosa (Clavicipitaceae; Hypocreales; Ascomycotina) has received special attention for medicinal purpose due to its various physiological activites. The nucleoside derivative N6-(2-hydroxyethyl) adenosine (HEA) isolated from it showed a $Ca^{2+}$ antagonistic effect and negative inotropic response. The artificial production of fruiting body of C. pruinosa has not been tried successfully yet by using living silkworm substrate. To develop techniques for the production of C. pruinosa stromata on a large scale, the infection of Bombyx mori with C. pruinosa and the growth characteristics of stroma of C. pruinosa were investigated. Also, studied about biological activities of fruiting body formed on silkworm. Infection rate of the silkworm pupae with C. pruinosa was the highest in injection inoculation. The formation of the fruiting body of C. pruinosa was quite good in the room controlled at $21{\sim}25^{\circ}C$, over 91% of relative humidity and over 1500 lx. Glucose concentration was high in the fruiting bodies of the silkworm pupae infected with C. pruinosa on a dry weight basis. The most abundant amino acid in the fruiting bodies was arginine and phenylalanine. The fruiting bodies of silkworm pupae infected with C. pruinosa was rich in oleic acid. The high amount of citric acid was found in the fruiting bodies of silkworm pupae infected with C. pruinosa.
This study was carried out to establish a proper cultivation site and diagnose the drought-tolerance of Aster scaber and Synurus deltoides leaves by using Pressure-volume curves. In order to measure pressure-volume (P-V) curves, Aster scaber and Synurus deltoides were cut off above ground part and the tip of the cutting were placed in water, which was covered with a plastic bag. Samples were kept overnight (about 12 hours) in darkness at room temperature (20~25$^{\circ}C$) to achieve maximal turgor (full saturation). The pressure in the chamber was gradually increased from 0.3MPa to 1.8MPa by nitrogen gas. After measured, leaf samples were dried at 80$^{\circ}C$ for 48 hours and dry weight of each samples were determined. The result of the original bulk osmotic potential at maximum turgor ${\Psi}^{sat}_o$ sat was lower -0.8 MPa in Aster scaber leaves than -0.7 MPa Synurus deltoides leaves. Also the osmotic potential at incipient plasmolysis ${\Psi}^{tlp}_o$ in Aster scaber leave was -0.9 MPa. In contrast, the value of maximum bulk modulus of elasticity $E_{max}$ of Aster scaber leaves were approximately two folds higher than that of Synurus deltoides leaves. The values of the relative water content at incipient plasmolysis $RWC^{tlp}$ are all above 90% showing that the function of osmoregulation is somewhat better, and Vo/DW, Vt/DW, Ns/DW of Synurus deltoides leaves were approximately 1~2 times higher than that of Aster scaber leaves. Thus, responses to water relations of Aster scaber and Synurus deltoides such as ${\Psi}^{sat}_o$, ${\Psi}^{tlp}_o$, $E_{max}$, ${\Psi}_{P,max}$, $RWC^{tl}$ were shown that the Aster scaber leaves was slightly higher drought-tolerance than Synurus deltoides leaves. However, in both of Aster scaber and Synurus deltoides, occurring incipient plasmolysis at the high water content, have a relatively lower drought-tolerance property indicating that growth of these plants are cultivated appropriate in high moisture soil sites.
Journal of The Korean Society of Grassland and Forage Science
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v.29
no.2
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pp.137-152
/
2009
Present experiment was conducted at the field of Gyeongnam Agricultural Research and Extension Services in Jinju city for two continuous cropping seasons to develop several adaptable and valuable year-round forage-producing system for elevating self-sufficiency and dollar-saving by reduced importing of crude forage. Twenty cropping systems were tested in experiment using whole crop barley (WCB), oat, rye, Italian ryegrass (IRG), and triticale in winter season and com, sorghum, sorghum ${\times}$ sudangrass hybrid, and oat in summer time. Sorghum ${\times}$ sudangrass hybrid showed highest fresh forage yield among experimented summer season crops, and followed com. Com produced the most dry matter yield, and followed sorghum${\times}$sudangrass hybrid, sorghum and oat in order. There was no significant effect of former winter crops on fresh and dry matter production succeeding summer time crops. Among winter season forage crops tested, oat showed the highest fresh and dry matter when clipped on mid-May, and followed triticale, IRG, rye and WCB. Winter-time cultivated crops showed no clear effect on the growth and forage (fresh and dry matter) producing ability of following summer crops. There was the most protein content in oat plant among summer season planted crops, and in sorghum for acid detergent fiber (ADF) and in sorghum ${\times}$ sudangrass hybrid for neutral detergent fiber (NDF), respectively. While, com showed highest value of relative feed value (RFV) and total digestive nutrients (TDN) among those crops. Among winter crops, the highest crude protein was in oat plant showing no significant differences of ADF and NDF, while, relatively higher value of RFV was recognized with rye and triticale. Also, triticale contained more TDN as compare to other forage crops. The cropping combinations such as com followed by (fb) rye and maize fb triticale were regarded as promising systems having higher dry matter producing ability among tested combinations. Considering TDN producing potential, the combinations with sorghum ${\times}$ sudangrass hybrid fb triticale andlor rye were would be suitable ones, coincidently. There was a tendency which elevating pH, electric conductivity (EC) and organic matter (OM) contents in soil after experiment comparing to before planting. More crude protein content in plant was shown at mid-May clipping as compared to the forage at April cut in all winter season grown crops. ADF and NDF contents were increased by delayed clipping showing decreased tendency of RFV and TDN in plant. In conclusion, many cropping systems would be available using above mentioned forage crops according to farmer's conditions and scale, etc.
To estimate the change in rice productivity around lake Juam due to construction of artificial lake, growth, yield components and yield of rice were measured at different locations around lake Juam for three years from 1994 to 1996. Automated weather stations(AWS) were installed nearby the experimental paddy fields, and daily maximum, average and minimum temperature, solar radiation, relative humidity, and precipitation were measured for the whole growing period of rice. Plant height, number of tillers, leaf area and shoot dry weight per hill were observed from 8 to 10 times in the interval of 7 days after transplanting. Yield and yield components of rice were observed at the harvest time. Simulation model of rice productivity used in the study was SIMRIW developed by Horie. The observed data of rice at 5 locations in 1994, 3 locations in 1995 and 4 locations in 1996 were inputted in the model to estimate the unknown parameters. Comparisons between observed and predicted values of shoot dry weights, leaf area indices, and rough rice yield were fairly well, so that SIMRIW appeared to predict relatively well the variations in productivity due to variations of climatic factors in the habitat. Climatic elements prior to as well as posterior to dam construction were generated at six locatons around lake Juam for thirty years by the method of Pickering et al. Climatic elements simulated in the study were daily maximum and minimum temperature, and amount of daily solar radiation. The change in rice productivity around lake Juam due to dam construction were estimated by inputting the generated climatic elements into SIMRIW. Average daily maximum temperature after dam construction appeared to be more or less lower than that before dam construction, while average daily minimum temperature became higher after dam construction. Average amount of daily solar radiation became lower with 0.9 MJ $d^{-1}$ after dam construction. As a result of simulation, the average productivity of habitats around lake Juam decreased about 5.6% by the construction of dam.
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