PI201232, a pepper line resistant to Phytophthora capsici Leonian, was crossed with PI2713222 and P1163192, resistant to Xanthomonas campestTis pv. vesicatoria (Doidge) Dye and inheritance of the resistance to each disease and genetic relationship between the two disease resistances was studied. Non-hypersensitive resistance to race 3 of X. c. pv. vesicatoria of PI271322 was inherited in a quantitative mode. Resitance to P. capsici of PI201232 was inherited in a mode close to two dominant alleles. Hypersensitive resistance to race 1 of X. c. pv. vesicatoria of PI271322 was inherited in a mode of single dominant allele. PI 163192 was consistently resistant to both race 1 and race 3 of X c. pv. vesicatoria and the resistance was inherited in a quantitative mode with high dominance effect. Resistance to X. c. pv. vesicatoria was inherited independently from resistance to P. capsici.
For the purpose of development of bioremediation technology for soil contaminated by chlorinated phenols, this study was focused on the isolation and characterization of bacteria capable of degrading chlorinated phenols, the establishment of analytical methods for chlorinated phenols, and the investigation of the contaminated sites. One site near the Incheon Industrial Complex was identified as a pentachlorophenol (PCP)-contaminated spot. The soil brought from the PCP-contaminated site contained 10-100$mu\textrm{g}$/g wet soil of PCP. Many bacterial strains capable of growing on a minimal medium containing PCP were isolated from 15 soil samples collected throughout the land, and among them, 10 active isolates were finally selected for the further studies on the biodegradability and for the use in in situ bioremediation of contaminated soil. These isolates showed species-specific pattern in PCP-decrease and cell growth in a minimal medium containing 500-1,000mg/ιPCP. Strain Bul degraded 90% of PCP at 216 hrs after incubation. Expecially, strain Bu34 was capable of degrading 4,000mg/ι PCP and was identified as Pseudomonas putida Bu34. It is seemed that the isolated active bacteria could be effectively used for the bioremediation of PCP-contaminated sites.
Bacterial black spot disease of prune fruit (Prunus salicina cv. formosa) has outbroke around major prune production area, Gimcheon, Euiseong and Gunwi in Gyungbuk province and has caused severe economic loss. Integrons PCR primer was designed along with sample pre-incubation and nested PCR method to enhance detection sensitivity for early detection of bacteria in fields. Designed integrons PCR primer successfully detected Xanthomonas arboricola pv. pruni from field collected samples, fruit, leaf, branch and even in raindrop collected from prune orchard. Pre-incubation along with nested PCR enhanced sensitivity to detect X. arboricola pv. pruni from seemingly healthy looking, symptomless branches. Designed integrons PCR can be used in prune nursery fields and in plant quarantine practice for the detection of X. arboricola pv. pruni.
Bacteriophages infecting Acidovorax citrulli, the causal agent of bacterial fruit blotch, have been proven to be effective for the prevention and control of this disease. However, the occurrence of bacteriophage-resistant bacteria is one of hurdles in phage biocontrol and the understanding of phage resistance in this bacterium is an essential step. In this study, we aim to investigate possible phage resistance of A. citrulli and relationship between phage resistance and pathogenicity, and to isolate and characterize the genes involved in these phenomena. A phage-resistant and less-virulent mutant named as AC-17-G1 was isolated among 3,264 A. citrulli Tn5 mutants through serial spot assays and plaque assays followed by pathogenicity test using seed coating method. The mutant has the integrated Tn5 in the middle of a cupin protein gene. This mutant recovered its pathogenicity and phage sensitivity by complementation with corresponding wild-type gene. Site-directed mutation of this gene from wild-type by CRISPR/Cas9 system resulted in the loss of pathogenicity and acquisition of phage resistance. The growth of AC-17-G1 in King's B medium was much less than the wild-type, but the growth turned into normal in the medium supplemented with D-mannose 6-phosphate or D-fructose 6-phosphate indicating the cupin protein functions as a phosphomannos isomerase. Sodium dodecyl sulfa analysis of lipopolysaccharide (LPS) extracted from the mutant was smaller than that from wild-type. All these data suggest that the cupin protein is a phosphomannos isomerase involved in LPS synthesis, and LPS is an important determinant of pathogenicity and phage susceptibility of A. citrulli.
Ground cherry (Physalis pubescens) is the most prominent species in the Solanaceae family due to its nutritional content, and prospective health advantages. It is grown all over the world, but notably in northern China. In 2019 firstly bacterial leaf spot (BLS) disease was identified on P. pubescens in China that caused by both BLS pathogens Xanthomonas euvesicatoria pv. euvesicatoria resulted in substantial monetary losses. Here, we compared whole genome sequences of X. euvesicatoria to other Xanthomonas species that caused BLS diseases for high similarities and dissimilarities in genomic sequences through average nucleotide identity (ANI) and BLAST comparison. Molecular techniques and phylogenetic trees were adopted to detect X. euvesicatoria on P. pubescens using recQ, hrpB1, and hrpB2 genes for efficient and precise identification. For rapid molecular detection of X. euvesicatoria, loop-mediated isothermal amplification, polymerase chain reaction (PCR), and real-time PCR techniques were used. Whole genome comparison results showed that the genome of X. euvesicatoria was more closely relative to X. perforans than X. vesicatoria, and X. gardneri with 98%, 84%, and 86% ANI, respectively. All infected leaves of P. pubescens found positive amplification, and negative controls did not show amplification. The findings of evolutionary history revealed that isolated strains XeC10RQ, XeH9RQ, XeA10RQ, and XeB10RQ that originated from China were closely relative and highly homologous to the X. euvesicatoria. This research provides information to researchers on genomic variation in BLS pathogens, and further molecular evolution and identification of X. euvesicatoria using the unique target recQ gene through advance molecular approaches.
Vegetables including Chinese cabbage and radish have been grown in alpine areas such as Muju, Namweon, Jinan and Jangsu during the shortage period of vegetables. The incidence of various diseases, environmental factors such as temperatures and rainfalls, and aphid populations as virus vector were observed at 10-day intervals from July to September in those areas. Disease incidence showed no significant difference among locations. Major diseases in Chin ese cabbage were bacterial soft rot, white spot, downy mildew, mosaic virus, Alternaria leaf spot and Fusarium seedling blight. Major diseases in radish were virus, white rust, black rot, root rot, leaf spot and Fusarium seedling blight. Disease incidence reached peak on Aug. 20 with $27\%$ infection in radish and with $20\%$ infection in Chinese cabbage during the growing season, and declined thereafter. Percentage of infection in each growth stage showed $25\%$ at root thickening stage and $26\%$ at the harvest time in radish; and the head formation stage, $24\%$ at the harvest time. The data indicate that disease incidence in radish increased rapidly at late growing stage and progressively increased in Chinese cabbage Seedling blight caused by Fusarium sp. and root rot caused by Aphanomyces sp. were also observed in those areas. Cool and wet weather appeared to be favorable for disease incidences during the rainy period of growing season although average temperature was about $25^{\circ}C$. Populations of aphids were lower in the alpine vegetable growing area than that of flat areas. Aphids as virus vectors from total aphids collected were $73.5\%$ or 289 virus vectors /993 total aphids in Namweon and $18.1\%$ or 31 virus vectors/171 total aphids in Muju. The most prerevalent species of aphids was Myzus persicae Sulz.
These experiments were conducted for decrease of injury by continuous cropping in the peanut fields of Chonbuk Wangkungarea. The continuous cropping field for four years was used in this experiment. Italian ryegrass and rye were cultivated andlime materials were distributed for improvement of soil fertility. The results were as follows; 1. Forage crops were cultivatedand lime materials were distributed on the continuous cropping field of peanut. The organic matter content of the expermentalplot cultivating Italian ryegrass was only 1.25%. The organic matter content of soil cultivated Italian ryegrass after distributedmagnesium lime was 1.37% and that of soil cultivated Italian ryegrass after distributed gypsum was 1.30%. It was highcontent comparing to that of soil distributed lime materials only. The organic matter content of soil cultivated rye after distributed gypsum was 1.77%. 2. The phosphate content of soil cutivated Italian ryegrass was 332ppm. The phosphate content ofsoil cultivated Italian ryegrass after distributed magnesium lime was 34Oppm and that of soil cultivated Italian ryegrass afterdistributed gypsum was 31 2ppm. The phosphate content of soil cultivated rye only was 386ppm. The phosphate content ofsoil cultivated rye after distributed gypsum was 41 8ppm. This phosphate content was lower than that of soil distributed limematerials only. 3. The phytotoxin content of soil cultivated Italian ryegrass after distributed magnesium lime was decreased to17.7% and that of soil cultivated Italian ryegrass after distributed gypsum was decreased to 25.3%. The phytotoxin content ofsoil cultivated rye after distributed magnesium lime was decreased to 12.0% and that of soil cultivated rye after distributedgypsum was decreased to 12.8% comparing to the phytotoxin content of soil distributed lime materials only. Italian ryegrasswas effective to decrease phytotoxin among the forage crops and gypsum was effective among the lime materials. 4. Abacterial wilt and a late spot of peanut which were known as, main reason of continuous cropping failure were surveyed.lnccidence of a bacterial wilt was 3.4% in the plot cultivated Italian ryegrass only and that was 2.9% in the plot cultivated ryeonly. lnccidence of a bacterial wilt was 2.5% in the plot cultivated Italian ryegrass after distributed magnesium lime and thatwas 2.3% in the plot cultivated rye after distributed gypsum. Inccidence plot cultivated forage crops was lower than that of plotdistributed lime materials. 5. Inccidence of a late spot was high in the plot cultivated forage crops ony, but it was low in the plotcultivated forage crops after distributed lime materials comparing to that of the control plot. 6. The growth and yield of peanutwere bad in the plot cultivated forage crops only comparing to the control plot distributed lime materials only. These resultswere same in the plot cultivated rye after distributed lime materials, but the growth and yield were grown up in the plotcultured Italian ryegrass after distributed lime materials.
The Bacillus subtilis pyrimidine biosynthetic (pyr) operon encodes all of the enzymes for the de novo biosynthesis of Uridine monophosphate (UMP) and additional cistrones encoding a uracil permease and the regulatory protein PyrR. The PyrR is a bifunctional protein with pyr mRNA-binding regulatory funtion and uracil phosphoribosyltransferase activity. To study the global regulation by the pyrR deletion, the proteome comparison between Bacillus subtilis DB104 and Bacillus subtilis DB104 ${\Delta}$pyrR in the minimal medium without pyrimidines was employed. Proteome analysis of the cytosolic proteins from both strains by 2D-gel electrophoresis showed the variations in levels of protein expression. On the silver stained 2D-gel with an isoelectric point (pI) between 4 and 10, about 1,300 spots were detected and 172 spots showed quantitative variations in which 42 high quantitatively variant proteins were identified. The results showed that production of the pyrimidine biosynthetic enzymes (PyrAA, PyrAB, PyrB, PyrC, PyrD, and PyrF) were significantly increased in B. subtilis DB104 ${\Delta}$pyrR. Besides, proteins associated carbohydrate metabolism, elongation protein synthesis, metabolism of cofactors and vitamins, motility, tRNA synthetase, catalase, ATP-binding protein, and cell division protein FtsZ were overproduced in the PyrR-deficient mutant. Based on analytic results, the PyrR might be involved a number of other metabolisms or various phenomena in the bacterial cell besides the pyrimidine biosynthesis.
In order to develop probiotics which may be a viable alternative of antibiotic use in pig industry, five bacterial strains (Lactobacillus sp. HN 52, 92, 98, 235 and AP 116) possessing antimicrobial properties were selected from 500 strains isolates of pig intestines. The bacteriocin produced by Lactobacillus sp. HN 235 displayed a relative broad spectrum of inhibitory activity against all Enterococcus strains, Pseudomonas aeruginosa, Listeria monocytogenes and Clostridium perfringens using the spot-on-lawn method. The production of antimicrobial substance started in the middle of the exponential growth phase, reached maximum levels (6,400 AU/mL) in the stationary phase, and then declined. Bacteriocin activity remained unchanged after 30 min of heat treatment at $95^{\circ}C$ and stable from pH 2.0 to 10 for 1 h, or exposure to organic solvents; however, it diminished after treatment with proteolytic enzymes. The molecular weight of the bacteriocin was about 5 kDa according to a tricine SDS-PAGE analysis.
Four Thai - rumen fistulated male swamp buffaloes (Bubalus bubalis), about four years old with $400{\pm}20kg$ liveweight, were randomly assigned according to a $4{\times}4$ Latin square design to receive dietary treatments. The treatments were: ground corn cob (GCC) replacement for cassava chip (CC) in concentrate at 0% (T1); GCC replacement at 33% (T2); GCC replacement at 67% (T3); and GCC replacement at 100% (T4), respectively. During the experiment, concentrate was offered at 0.5% BW while 5% urea-treated rice straw was given at ad libitum. The result revealed that there was no effect of GCC replacement on DMI among treatments. In addition, digestibilities of DM, OM and CP were not different while aNDF linearly increased with an increasing level of GCC replacement. However, GCC replacement did not affect rumen fermentation such as ruminal pH, $NH_3$-N and VFA concentration; except C3 proportion which was the highest at 33% replacement while the lowest was at 100% replacement. All replacements of GCC resulted in similar protozoal and bacterial populations and microbial protein synthesis (MPS). Purine derivatives (PD) concentration in urine and PD to creatinine (PDC) index were varied with time of urination and among treatments at 0 to 8 and 8 to 16 h post feeding and higher values were shown among the GCC replacement groups. However at 16 to 24 h-post feeding, it was untraceable. In addition, creatinine concentration was similar among all treatments at every sampling time. Based on the above results, GCC can be used as an energy source for swamp buffalo fed with rice straw. Spot sampling of urine can be used for purine derivatives determination.
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