• Mycobiology
• /
• 제37권3호
• /
• pp.225-229
• /
• 2009

Single spore isolates of Plasmodiophora brassicae e4 and e9 obtained from diseased Chinese cabbage were identified as race 4 and race 9, respectively, by the Williams' differential variety set. To confirm the possibility of variation in same generation and progeny of a single spore isolate of P. brassicae, random amplified polymorphic DNA (RAPD) analysis was conducted using the URP 3, 6 and OPA 7 primers. There was no difference in band type at each part of the gall of Chinese cabbage obtained by inoculation of e4 and e9 and amplification using the URP 3 and 6 primers when the same generation was analyzed. In addition, the progeny analysis, which was expanded to the third generation and conducted using the URP 3 and OPA 7 primers, revealed no differences in the band type of the e4 isolate. Based on these results, the single spore isolate of P. brassicae was genetically stable.

• Journal of Microbiology and Biotechnology
• /
• 제11권1호
• /
• pp.72-78
• /
• 2001

It is possible ot prepare protoplasts of the zygomycete fungus, Phycomyces blakesleeanus, by digesting the cell wall of spore germlings with commercially available chitinase and chitosanase. However, the cells without any cell walls immediately form large aggregates, and thus, it is difficult to isolate the individually separated protoplasts. Inherent problem with the formation of aggregates in preparing protoplasts could be solved by the use of bovine serum albumin (BSA). As a result, we were able to prepare a large number of single protoplsts quickly and easily. We took time-lapse photomicrographs of the formation of protoplasts, and found that there were certain regions of the cell wall of spore germlings that were sensitive to chitinase and chitosanase, although the cell wall of the original spores is known to be insensitive to these enzymes. There are two kinds of cell walls on a spore germling; one with a bound wheat germ agglutinin (WGA), and the other a bound concanavalin A (ConA). Furthermore, only cells with walls which had bound WGA were able to regenerate, while those with walls with bound ConA were not able to regenerate.

• Journal of Crop Science and Biotechnology
• /
• 제11권2호
• /
• pp.101-106
• /
• 2008

To identify inheritance of clubroot disease resistance genes in Chinese cabbage, seedling tests of $BC_1P_1,\;BC_1P_2$, and $F_2$ populations derived from $F_1$ hybrid(var. CR Saerona) using single spore isolate(race 4 identified with William's differential host) from Plasmodiophora brassciae were conducted. Resistance(R) and susceptible(S) plants segregated to 1:0 in backcross to the resistant parent. The $F_2$ population segregated in a 3(R):1(S) ratio. This result implied that the resistance of clubroot disease is controlled by a single dominant gene to the race 4 of P. brassicae in CR Saerona. To develop DNA markers linked to clubroot resistance genes, 185 plants of CR Saerona among $F_2$ populations were used. A total of 300 arbitrary decamer was applied to $F_2$ population using BSARAPD(Bulked segregant analysis-Randomly amplified polymorphic DNA). One RAPD marker linked to clubroot resistance gene in CR Saerona($OPJ_{1100}$) was identified. This marker was 3.1 cM in distance from resistance gene in $F_2$ population. This marker may be useful for a marker-assisted selection(MAS) and gene pyramiding of the clubroot disease resistant gene in Chinese cabbage breeding programs.

• 한국균학회지
• /
• 제9권3호
• /
• pp.133-139
• /
• 1981

양송이 21계통(系統)으로부터 포자(胞子)를 채취(採取)하여 247개(個)의 단(單), 다포자(多胞予) 발아균주(發芽菌株)를 분리(分離)한 후(後) 생산력(生産力) 예비(豫備) 시험(試驗)과 균총(菌叢)의 형태(形態) 및 생장(生長)에 관(關)한 실내(室內) 시험(試驗)에서 4개(個) 균주(菌株)를 우수계통(優秀系統)으로 선발(選拔)하여 현(現) 장려 품종(品種)인 505호(號) 및 703 호(號)와 생산력(生産力)을 비교(比較)한 결과(結果) 705호(號)는 수량(收量), 품질등(品質等) 제특성(諸特性)이 가장 우수(優秀)하였으며 그 특성(特性)은 다음과 같다. 1. $1979{\sim}'80$의 생산력(生産力) 검정(檢定) 결과(結果) t신계통(新系統) 705호(號)은 크림색(色) 계통(系統)으로 수량(收量)은 703호(號)보다 13% 증수(增收)되고 자숙수율(煮熟收率)이 높고 마이코곤에도 중도(中度)의 저항성(抵抗性)이었다. 2. 705호(號)는 균사생장(菌絲生長) 적온(適溫)이 $25^{\circ}C$이나 $15^{\circ}C$의 저온(低溫)에서도 다른 균주(菌株)보다는 비교적(比較的) 생장(生長)이 양호(良好)하였고 최적(最適) 퇴비(堆肥) 수분(水分) 함량(含量)은 65%, 복토(覆土) 최적(最適) 산도(酸度)는 pH 7.8 내외(內外)였다. 3. 신계통(新系統) 705호(號)은 703호(號)에 비(比)하여 버섯의 발생수(發生數)가 많으며 자실체(子實體)의 크기 및 부위별(部位別) 크기 비율(比率)은 505호(號)와 703호(號)의 중간(中間) 정도(報度)이다.

• 식물병연구
• /
• 제13권1호
• /
• pp.45-49
• /
• 2007

국내의 여러 배추 생산지로부터 뿌리혹을 채집하여 단포자 균주(SSI)를 분리하였다. 분리한 단포자를 접종한 후 총 321개의 뿌리를 수거하여 병징 발현 여부를 관찰하였다. 접종한 개체 중 단 한 개체에서만 작은 혹이 형성되었고, 70개 개체에서 미약한 기형 증상을 보였다. 이들 이상 증상을 보인 개체를 마쇄하여 단포자 균주의 안정적 증식을 위한 접종원으로 사용하였다. 단포자를 접종하여 발병시킨 개체로부터 얻은 포자들을 재접종하였을 때가 1차 단포자 접종하였을 때보다 병징이 훨씬 확대되었고, 이후 실험에 사용할 수 있을 정도로 증식된 개체는 15균주였다. 이 15균주들을 Williams의 판별기주에 접종하여 분석하였을 때, race 1이 4균주, race 4는 7균주, race 9는 2균주, race 11이 2균주로 각각 판별되어, race 4가 국내에 가장 우점하는 것으로 밝혀졌다. 이들 race들은 모두 배추, 무, 순무에 단포자 접종하여 얻어진 균주들로, 병원성 검정에서도 4종의 판별품종 중 Rutabaga 계열(스페인산 순무)에 속하는 'Laurentian'과 'Wilhelmsburger' 두 품종에만 병원성을 보였다. 따라서 이 race들이 배추, 무, 순무에 대해 강한 병원성을 보이는 것으로 보아 국내의 P. brassicae 군집은 배추와 무, 순무 등에 강한 병원성을 나타내는 race가 우점하는 것으로 판단된다. 이는 이들 작물이 병 발생량에 따른 국내 십자화과 채소 감수 비율과도 연관이 있으리라 생각된다.

• Journal of Microbiology and Biotechnology
• /
• 제20권2호
• /
• pp.410-414
• /
• 2010

A bacterial strain, 13-$25^T$ with xylanolytic activity isolated from a single soil sample, was characterized with respect to its phenetic and phylogenetic characteristics. The cells of the isolate are Gram-staining variable rods, but spore formation was not observed. This strain is catalase- and oxidase-positive, and able to degrade starch and xylan. The predominant fatty acids are anteiso-$C_{15:0}$, $C_{16:0}$, and iso-$C_{16:0}$. The major respiratory quinone is menaquinone 7(MK-7), with a polar lipid profile consistent with the genus Cohnella. The DNA G+C content is 54.3 mol%. The 168 rRNA gene sequence analysis indicates that this organism belongs to the genus Cohnella, with Cohnella panacarvi as the closest phylogenetic neighbor. Low levels of 168 rRNA gene sequence similarity (<97.0%) with respect to other taxa with published names and the identification of distinctive phenetic features in the isolate indicate that the strain 13-$25^T$ represents a novel species of the genus Cohnella, for which the name Cohnella damensis sp. novo is proposed. The type strain is 13-$25^T$ (=CCTCC AB $208103^T$=KCTC $13422^T$).

• Mycobiology
• /
• 제46권3호
• /
• pp.278-282
• /
• 2018

Chrysanthemum coronarium is an economically important plant in Asia, and used medicinally, ornamentally and as a vegetable. In April 2017, leaf spot disease on C. coronarium was observed in Shiyan, Hubei, China. A single-spore isolate was obtained and identified based on morphology and sequence analysis using four regions (rDNA ITS, GAPDH, $EF-1{\alpha}$, and RPB2). The results indicated that the fungus is Alternaria argyranthemi. The pathogenicity tests revealed that the species could cause severe leaf spot and blight disease on the host. This is the first report of leaf spot disease on C. coronarium caused by A. argyranthemi in the world, which is also a new record of Alternaria species in China.

• 한국작물학회지
• /
• 제45권2호
• /
• pp.143-149
• /
• 2000

From the soils of soybean fields in Cotton Branch Station (CBS) and Pine Tree Station (PTS), Arkansas, USA, various single spore isloates of sudden death syndrome (SDS) pathogen were obtained on modified Nash ＆ Snyder's medium (MNSM) with dilution plating technique and transferred to potato dextrose agar (PDA) medium to identify the cultural colony shape. The colony shapes of these isolates resembled F. solani isolate 171 which was white and chalky shaped on MNSM and most of them had unique form of morphology which produced white margin and blue center colony on PDA. Although, some of these isolates had more dark blue or showed slightly different color, all isolates that were selected randomly for green-house inoculation assay produced typical foliar symptoms on leaves of soybean, Hartz 6686. To determine the genetic differences among the isolates, mitochondrial DNA restriction fragment length polymorphism (RFLP) was conducted with fourty isolates from both fields, using mtDNA probes, 2U18 and 4U40, derived from Colletotrichum orbiculare. We obtained distinctive RFLPs in each treatment of restriction enzyme, EcoRI and HaeⅢ. Isolates, 11-2-5 and 14-3-1-1, from CBS and isolates, 104-3-1-2 and 701-1-5-1, from PTS showed different band patterns from 171 in both or in either treatment of restriction enzymes. Even if some of these isolates showed heterogeneous, they were more closer to 171 than PN603. And, also, rest of the thirty-six isolates had exactly same polymorphisms as 171 in each treatment of restriction enzyme. Although, some of the isolates showed the different morphological shape on PDA and slightly different band patterns on RFLPs, all of the isolates selected on MNSM due to their distinctive colony shape from other fungi produced the typical foliar symptoms on soybean leaves in greenhouse inoculation assay. It might be suggested that these isolates were not genetically different from check isolate 171 and they were unique strain of F. solani.

• The Plant Pathology Journal
• /
• 제26권3호
• /
• pp.216-222
• /
• 2010

Fusarium proliferatum is the causal agent of stalk and root rot disease of maize, foot rot disease of rice, basal and root rot disease of onion and knife cut disease of sugarcane in Iran. In recent years, incidence and severity of these diseases have been increased in Iran. Fifty seven F. proliferatum single-spore isolates collected from diseased maize, rice, onion and sugarcane plants at different areas were used to study genetic diversity by determination of vegetative compatibility groups (VCGs). Chlorate-resistant nitrate non-utilizing (nit) mutants were recovered from selected isolates of F. proliferatum and used in complementation tests. All isolates in which both nit1 and NitM (or nit3) mutants were recovered, demonstrated self-compatibility. Vegetative compatibility tests by pairing nit mutants identified 30 VCGs among 57 isolates. Twenty-three isolates belonged to singlemember VCGs and the remaining 34 isolates, belonged to other seven multimember VCGs. Segregation of F. proliferatum isolates obtained from various area and host plants into different VCGs in Iran is reported for the first time. In this study, none of isolates obtained from rice complemented with any other isolates from onion and sugarcane and, non complementation occurred between onion and sugarcane isolates. Also, only one complementation occurred between one isolate of maize and one isolate of sugarcane and rice. Thus, a correlation between VCGs grouping and host preferences was founded. It is concluded that natural populations of F. proliferatum in Iran are probably genetically divergent and include isolates representing a potential risk for disease development.

• 미생물학회지
• /
• 제51권4호
• /
• pp.419-426
• /
• 2015

가축산업분야에서 항생제의 사용이 금지됨에 따라, 질병 예방을 통한 축산농가의 생산성 향상을 위해 사료첨가제인 미생물제재의 개발과 같은 예방적 수단이 필요하게 되었다. 본 연구는 가축의 생산성을 높이기 위해 사료 분해 능력이 좋고 항균활성이 뛰어난 포자 형성 Bacillus 균주 3종인 B. sutilis LCB7, B. licheniformis SHS14, B. amyloliquefaciens LCB10을 우수 균주로 선발하였다. 최종적으로 선발한 Bacillus 3종을 1:1:1 비율로 혼합하여 혼합 종균을 제조하여 항균시험(in vitro) 결과, 단일 3종 및 lincomycin과 비교하여 유사한 활성을 보여주었으며, 송아지를 이용하여 항균활성 시험(in vivo)을 실시한 결과에서도 lincomycin 투여 대비 90% 수준의 높은 활성을 보여주었다. 개발한 혼합 종균의 안정적 보존을 위해 혼합 종균을 제조하여 다시 증균을 통하여 미생물군집 분석을 통하여 확인한 결과, 초기 군집비율과 증균 후 군집비율이 매우 유사하게 유지되었다. 이로서 본 연구에서는 선발된 Bacillus 균주 3종을 이용하여 제조한 혼합 종균이 사료첨가제용 미생물제재로서 이용 가능함을 최종 확인하였다.