Fusarium prolifertum KGL0401의 지베렐린 생산 최적조건과 waito-c 생장에 미치는 영향

김종국,임순옥,이진형,이인중,이인구 한국생명과학회 2007 생명과학회지 Vol.17 No.2

Fusarium proliferatum KGL0401 was previously isolated from Physalis alkekengi var. francheti plant roots and exhibited higher GA productivity than wild type Gibberella fujikuroi. The aim of this work was to find out an optimal culture condition for GA production. Various carbon(fructose, glucose, lactose, maltose, sucrose) and nitrogen(KNO3, urea, glycine, NaNO3, NH4Cl) sources were used for this study. GAs activities were analysed by gas chromatography and mass spectrometry(GC-MS). The highest yield of GA3 was found in the growth medium supplemented with sucrose as carbon source and NH4Cl as nitrogen source. The optimum carbon-nitrogen concentration for GA3 production was found to be 0.5 M:0.17 M. Supernatant was prepared from the culture fluid of F. proliferatum KGL0401 cultured for 7 days at 30℃ and the 10 ul supernatant was treated with 2 leaf-rice seedling. 본 연구는 지베렐린을 생산하는 곰팡이로 알려진 야생균주 Gibberella fujikuroi 보다 더 많은 지베렐린을 생산하는 균인 Fusarium proliferatum KGL0401를 꽈리 뿌리에서 분리하였으며[13], 지베렐린 생산을 위한 최적 탄소원과 질소원의 종류, C:N ratio에 대해서 실험을 수행하였다. 지베렐린 중 생물학적 활성이 가장 높은 GA3를 가장 많이 생산하는 탄소원은 sucrose(7.02 ng/ml)이었으며, 질소원은 NH4Cl(187.63 ng/ml)이었다. 그리고 최적 C:N ratio를 찾기 위해 탄소원(0 - 1.5 M)과 질소원(0 - 0.47 M)을 배지에 첨가하였다. 결과적으로 최적 탄소원과 질소원의 ratio가 0.5 M : 0.17 M일 때 생물학적 활성을 가진 GA3를 140.0 ng/ml로 가장 많이 생산하는 것으로 나타났다.그리고 bioassay 결과 GA1, GA3 GA4과 GA7의 함량이 가장 높았던 C:N ratio가 0.5 M : 0.17 M 일 때의 배양액 10 ul을 처리한 waito-c 볍씨의 길이가 평균 11.1 cm로 가장 높게 나타났다.

Fusarium prolifertum KGL0401의 지베렐린 생산 최적조건과 waito-c 생장에 미치는 영향

임순옥,이진형,이인중,이인구,김종국,Rim, Soon-Ok,Lee, Jin-Hyung,Lee, In-Jung,Rhee, In-Koo,Kim, Jong-Guk 한국생명과학회 2007 생명과학회지 Vol.17 No.1

본 연구는 지베렐린을 생산하는 곰팡이로 알려진 야생균주 Gibberella fujikuroi보다 더 많은 지베렐린을 생산하는 균인 Fusarium proliferatum KGL0401를 꽈리 뿌리에서 분리하였으며[13], 지베렐린 생산을 위한 최적 탄소원과 질소원의 종류, C:N ratio에 대해서 실험을 수행하였다. 지베렐린 중 생물학적 활성이 가장 높은 $GA_3$를 가장 많이 생산하는 탄소원은 sucrose(7.02 ng/ml)이었으며, 질소원은 $NH_4Cl$(187.63 ng/ml)이었다. 그리고 최적 C:N ratio를 찾기 위해 탄소원(0 - 1.5 M)과 질소원(0 - 0.47M)을 배지에 첨가하였다. 결과적으로 최적 탄소원과 질소원의 ratio가 0.5 M : 0.17 M일 때 생물학적 활성을 가진 $GA_3$를 140.0 ng/ml로 가장 많이 생산하는 것으로 나타났다. 그리고 bioassay 결과 $GA_1,\;GA_3\;GA_4$과 $GA_7$의 함량이 가장 높았던 C:N ratio가 0.5 M : 0.17 M 일 때의 배양액 10 ul을 처리한 waito-c 볍씨의 길이가 평균 11.1 cm로 가장 높게 나타났다. Fusarium proliferatum KGL0401 was previously isolated from Physalis alkekengi var. francheti plant roots and exhibited higher GA productivity than wild type Gibberella fujikuroi. The :tim of this work was to find out an optimal culture condition for GA production. Various carbon(fructose, glucose, lactose, maltose, sucrose) and nitrogen($KNO_3$, urea, glycine, $NaNO_3,\;NH_4Cl$) sources were used for this study. GAs activities were analysed by gas chromatography and mass spectrometry(GC-MS). The highest yield of $GA_3$ was found in the growth medium supplemented with sucrose as carbon source and $NH_4Cl$ as nitrogen source. The optimum carbon-nitrogen concentration for $GA_3$ production was found to be 0.5 M:0.17 M. Supernatant was prepared from the culture fluid of F. proliferatum KGL0401 cultured for 7 days at 3 0'E and the 10 ul supernatant was treated with 2 leaf-rice seedling.

Molecular mapping of <i>qBK1</i> ^<i>WD</i> , a major QTL for bakanae disease resistance in rice

Lee, Sais-Beul,Hur, Yeon-Jae,Cho, Jun-Hyeon,Lee, Jong-Hee,Kim, Tae-Heon,Cho, Soo-Min,Song, You-Chun,Seo, Young-Su,Lee, Jungkwan,Kim, Tae-sung,Park, Yong-Jin,Oh, Myung-Kyu,Park, Dong-Soo Springer US 2018 Rice Vol.11 No.-

<P><B>Background</B></P><P>Bakanae or foot rot disease is a prominent disease of rice caused by <I>Gibberella fujikuroi</I>. This disease may infect rice plants from the pre-emergence stage to the mature stage. In recent years, raising rice seedlings in seed boxes for mechanical transplanting has increased the incidence of many seedling diseases; only a few rice varieties have been reported to exhibit resistance to bakanae disease. In this study, we attempted to identify quantitative trait loci (QTLs) conferring bakanae disease resistance from the highly resistant <I>japonica</I> variety Wonseadaesoo.</P><P><B>Results</B></P><P>A primary QTL study using the genotypes/phenotypes of the recombinant inbred lines (RILs) indicated that the locus <I>qBK1</I><SUP><I>WD</I></SUP> conferring resistance to bakanae disease from Wonseadaesoo was located in a 1.59 Mb interval delimited on the physical map between chr01_13542347 (13.54 Mb) and chr01_15132528 (15.13 Mb). The log of odds (LOD) score of <I>qBK1</I><SUP><I>WD</I></SUP> was 8.29, accounting for 20.2% of the total phenotypic variation. We further identified a gene pyramiding effect of two QTLs, <I>qBK</I><SUP><I>WD</I></SUP> and previously developed <I>qBK1</I>. The mean proportion of healthy plant for 31 F<SUB>4</SUB> RILs that had no resistance genes was 35.3%, which was similar to that of the susceptible check variety Ilpum. The proportion of healthy plants for the lines with only <I>qBK</I><SUP><I>WD</I></SUP> or <I>qBK1</I> was 66.1% and 55.5%, respectively, which was significantly higher than that of the lines without resistance genes and that of Ilpum. The mean proportion of the healthy plant for 15 F<SUB>4</SUB> RILs harboring both <I>qBK</I><SUP><I>WD</I></SUP> and <I>qBK1</I> was 80.2%, which was significantly higher than that of the lines with only <I>qBK</I><SUP><I>WD</I></SUP> or <I>qBK1</I>.</P><P><B>Conclusion</B></P><P>Introducing <I>qBK</I><SUP><I>WD</I></SUP> or pyramiding the QTLs <I>qBK</I><SUP><I>WD</I></SUP> and <I>qBK1</I> could provide effective tools for breeding rice with bakanae disease resistance. To our knowledge, this is the first report on a gene pyramiding effect that provides higher resistance against bakanae disease.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (10.1186/s12284-017-0197-7) contains supplementary material, which is available to authorized users.</P>

벼 상자육묘에서 규산코팅볍씨의 건묘육성과 벼키다리병 경감효과

강양순,김완중,노재환 한국작물학회 2017 한국작물학회지 Vol.62 No.1

We investigated the effect of silicate coating of rice seeds on bakanae disease incidence and the quality of seedlings raised in seedling boxes and transplanted into pots. The silicate-coated rice seed (SCS) was prepared as follows. Naturally infested rice seeds not previously subjected to any fungicidal treatment were dressed with a mixture of 25% silicic acid at pH 11 and 300-mesh zeolite powder at a ratio of 50 g dry seed - 9 mL silicic acid - 25 g zeolite powder. The following nursery conditions were provided : Early sowing, dense seeding in a glass house with mulching overnight and no artificial heating, which were the ideal conditions for determining the effect on the seed. The nursery plants were evaluated for Gibberella. fujikuroi infection or to determine the recovery to normal growth of infected nursery plants in the Wagner pot. Seedlings emerged 2-3 days earlier for the SCS than they did for the non-SCS control, while damping-off and bakanae disease incidence were remarkably reduced. Specifically, bakanae disease incidence in the SCS was limited to only 7.8% for 80 days after sowing, as compared to 91.6% of the non-SCS control. For the 45-days-old SCS nursery seedlings, the fresh weight was increased by 11% and was two times heavier, with only mild damage compared to that observed for non-SCS. Even after transplanting, SCS treatment contributed to a lower incidence of further infections and possibly to recovery of the seedlings to normal growth as compared to that observed in symptomatic plants in the pot. The active pathogenic macro-conidia and micro-conidia were considerably lower in the soil, root, and seedling sheath base of the SCS. In particular, the underdeveloped macro-conidia with straight oblong shape without intact septum were isolated in the SCS ; this phenotype is likely to be at a comparative etiological disadvantage when compared to that of typical active macro-conidia, which are slightly sickle-shaped with 3-7 intact septa. A active intact conidia with high inoculum potential were rarely observed in the tissue of the seedlings treated only in the SCS. We propose that promising result was likely achieved via inhibition of the development of intact pathogenic conidia, in concert with the aerobic, acidic conditions induced by the physiochemical characteristics associated with the air porosity of zeolite, alkalinity of silicate and the seed husk as a carbon source. In addition, the resistance of the healthy plants to pathogenic conidia was also important factor.

Fine mapping of <i>qBK1</i> , a major QTL for bakanae disease resistance in rice

Lee, Sais-Beul,Kim, Namgyu,Hur, Yeon-Jae,Cho, Su-Min,Kim, Tae-Heon,Lee, Ji-youn,Cho, Jun-Hyeon,Lee, Jong-Hee,Song, You-Chun,Seo, Young-Su,Ko, Jong-Min,Park, Dong-Soo Springer US 2019 Rice Vol.12 No.-

<P><B>Background</B></P><P>Bakanae disease is an important fungal disease caused by <I>Gibberella fujikuroi</I>. Incidence of rice bakanae disease creates serious problems in the foremost rice growing countries, and no rice variety has been found to be completely resistant to this disease. However, breeding rice varieties resistant to bakanae disease may be a cost-saving solution preferable to the application of fungicides. In this study, we aimed to determine the exact position and the candidate gene for <I>qBK1</I>, a major resistant quantitative trait locus (QTLs) for bakanae disease.</P><P><B>Results</B></P><P>The genotypes/phenotypes of recombinants selected from backcrossed recombinant inbred lines of two rice varieties, Shingwang (resistant) and Ilpum (susceptible), indicated that the locus <I>qBK1</I>, conferring resistance to bakanae disease in Shingwang, was delimited to a 35-kb interval delimited by InDel 18 (23.637 Mbp) and InDel 19–14 (23.672 Mbp). Sequence analysis of this 35-kb region revealed four candidate genes, LOC_Os01g41770, LOC_Os01g41780, LOC_Os01g41790, and LOC_Os01g41800. There were many non-synonymous SNPs in LOC_Os01g41770 and the transcript of LOC_Os01g41790 was early terminated in Shingwang, whereas there were no differences in both LOC_Os01g41780 and LOC_Os01g41800 sequences between Ilpum and Shingwang. Expression profiling of the four candidate genes showed the up-regulation of LOC_Os01g41770, LOC_Os01g41780, and LOC_Os01g41790 in Ilpum and of LOC_Os01g41800 in Shingwang after inoculation of <I>G. fujikuroi.</I></P><P><B>Conclusion</B></P><P>Utilization of marker-assisted selection (MAS) with a precise molecular marker on <I>qBK1</I> could provide an effective tool for breeding rice varieties resistant to bakanae disease. To our knowledge, this is the first report on fine mapping and candidate gene approaches for identifying the gene for <I>qBK1</I>.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (10.1186/s12284-019-0295-9) contains supplementary material, which is available to authorized users.</P>

Bioprocess Strategies and Recovery Processes in Gibberellic Acid Fermentation

Shukla, Ruchi,Srivastava, Ashok K.,Chand, Subhash The Korean Society for Biotechnology and Bioengine 2003 Biotechnology and Bioprocess Engineering Vol.8 No.5

Gibberellic acid (GA$_3$) is a commercially important plant growth hormone, which is gaining much more attention all over the world due to its effective use in agriculture and brewing industry. Industrially it is produced by submerged fermentation technique using Ascomycetous fungus Gibberella fujikuroi. Solid state and immobilized cell fermentation techniques had also been developed as an alternative to obtain higher yield of GA$_3$. This review summarizes the problems of GA$_3$ fermentation such as production of co-secondary metabolites along with GA$_3$, substrate inhibition and degradation of GA$_3$ to biologically inert compound gibberellenic acid, which limits the yield of GA$_3$ in the fermentation medium. These problems can be overcome by various bioprocessing strategies e.g. two - stage and fed batch cultivation processes. Further research on bioreactor operation strategies such as continuous and / or extractive fermentation with or without cell recycle / retention system need to be investigated for improvement in yield and productivity. Down stream processing for GA$_3$ isolation is also a challenge and procedures available for the same have been critically evaluated.

Gibberella fujikuroi ATCC 12616으로 부터 얻어진 변이주 Gibberella fujikuroi G-36의 Gibberellic Acid의 배양조건

오영준 한국산업미생물학회 2000 한국미생물·생명공학회지 Vol.28 No.3

곰팡이 Gibberella fujikuroi ATCC12616을 mutagen, N-methyl-N^1-nitro-N-nitrosoguanidine과 hydroxylamine으로 처리하여 지베렐린 생산 수률이 가장 높은 변이주 G. fujikuroi G-36을 선별하였다. 변이주 G. fujikuroi G-36는 모균주 G. fujikuroi ATCC 12616과 비교하여 볼 때 지베렐린 생산수율이 약 32%향상된 것으로 관찰되었다. 이 균주를 액체배지에서 발효시키면 주 탄소원으로 sucrose를 사용할 때 높은 생산성을 나타내었으며 초기배지 pH와 온도는 각각 pH 4.0, 20℃ 조건이 가장 좋았다. 배양온도를 30℃에서 3일 배양후 20℃로 낮추어 7일간 배양하면 130㎎/l의 가장 높은 지베렐린이 생성됨을 알았다. A mutant Gibberella fujikuroi G-36 was selected by metagenesis of Gibberella fujikuroi ATCC 12616 with mutagens such as N-methyl-N'-nitro-N-nitrosoguanidine and hydroxylamine for improving productivity of gibberellic acid. The mutant strain produced gibberellic acid (78㎎/l) more than that of wilde type. A fermentation medium containing glucose, NH_4NO_3, MgSO_4, KH_2PO_4 and trace elements was developed for the maximal production of a gibberellic acid by the mutant. The fluctuating cultural temperature that was varied from 30℃ to 20℃ resulted in higher GA yield than that of fixed cultural temperature at 28℃.

RESEARCH ARTICLES : Incidence, Molecular Characteristics and Pathogenicity of Gibberella fujikuroi Species Complex Associated with Rice Seeds from Asian Countries

( Young Ah Jeon ),( Seung Hun Yu ),( Young Yi Lee ),( Hong Jae Park ),( Sokyoung Lee ),( Jung Sook Sung ),( Yeon Gyu Kim ),( Ho Sun Lee ) 한국균학회 2013 Mycobiology Vol.41 No.4

Gibberella fujikuroi species complex (GFSC) was isolated from rice (Oryza sativa L.) seed samples from ten Asian countries and investigated for incidence of GFSC, molecular characteristics, and pathogenicity. Regardless of geographic origin, GFSC was detected with incidences ranging from 3% to 80%. Four species, Fusarium fujikuroi, F. concentricum, F. proliferatum, and F. verticillioides, were found to show an association with rice seeds, with F. fujikuroi being the predominant species. In phylogenetic analyses of DNA sequences, no relationship was found between species, isolates, and geographic sources of samples. Unidentified fragments of the β-tubulin gene were observed in ten isolates of F. fujikuroi and F. verticillioides. With the exception of three isolates of F. fujikuroi, F. fujikuroi, F. proliferatum, and F. verticillioides were found to have FUM1 (the fumonisin biosynthetic gene); however, FUM1 was not found in isolates of F. concentricum. Results of pathogenicity testing showed that all isolates caused reduced germination of rice seed. In addition, F. fujikuroi and F. concentricum caused typical symptoms of bakanae, leaf elongation and chlorosis, whereas F. proliferatum and F. verticillioides only caused stunting of seedlings. These findings provide insight into the characteristics of GFSC associated with rice seeds and might be helpful in development of strategies for management of bakanae.

우리나라 벼와 옥수수로부터 분리한 Gibberella fujikuroi 종복합체와 Fusarium commune 소속 균주의 푸모니신 생성능

이수형,김지혜,손승완,이데레사,윤성환 한국식물병리학회 2012 식물병연구 Vol.18 No.4

Gibberellea fujikuroi species (Gf) complex comprises at least 15 species, most of which not only causes serious plant diseases, but also produces mycotoxins including fumonisins. Here, we focused on the abilities of the field isolates belonging to the Gf complex associated with rice and corn, respectively in Korea to produce fumonisin, all of which were confirmed to carry FUM1, the polyketide synthase gene essential for fumonisin biosynthesis. A total of 88 Gf complex isolates (55 F. fujikuroi, 10 F. verticillioides, 20 F. proliferatum, 2 F. subglutinans, and 1 F. concentricum), and 4 isolates of F. commune, which is a non-member of Gf complex,were grown on rice substrate and determined for their production levels of fumonisins by a HPLC method. Most isolates of F. verticillioides and F. proliferatum, regardless of host origins, produced fumonisin B1 and B2at diverse ranges of levels (0.5−2,686.4 μg/g, and 0.7−1,497.6 μg/g, respectively). In contrast, all the isolates of F. fujikuroi and other Fusarium species examined produced no fumonisins or only trace amounts (<10 μg/g)of fumonisins. Interestingly, the frequencies of relatively high fumonisin-producers among the F. proliferatum and F. fujikuroi isolates derived from corn were higher than those among the fungal isolates from rice. In addition, it is a first report demonstrating the ability of the FUM1-carrying F. commune isolates from rice to produce fumonisins.

Population Structure of the Gibberella fujikuroi Species Complex Associated with Rice and Corn in Korea

Kim, Ji-Hye,Kang, Mi-Ran,Kim, Hee-Kyoung,Lee, Seung-Ho,Lee, Theresa,Yun, Sung-Hwan The Korean Society of Plant Pathology 2012 Plant Pathology Journal Vol.28 No.4

Several species belonging to the Gibberella fujikuroi species (Gf ) complex are commonly associated with rice and corn, not only causing serious diseases, but also producing fumonisins, a group of mycotoxins harmful to animals and humans. To characterize the population structure of the putative fumonisin-producing Gf complex in Korea, we obtained 276 candidate isolates from rice and corn harvested in 2009 and 2010 by diagnostic polymerase chain reaction with several specific primer sets. Phylogenetic trees were constructed using multilocus sequences (combined RPB2 and EF1A, totaling 1.6 kb) from these isolates. Among the 135 isolates from rice, F. fujikuroi (teleomorph: G. fujikuroi; 59.3%) and F. proliferatum (G. intermedia; 13.3%) were predominant, followed by F. concentricum (5.9%). Additionally, twenty-five (18.5%) rice isolates belonged in a distinct subclade of F. commune, a non-member of the Gf complex. In contrast, F. verticillioides was the most predominant species (38.3%) among the 141 corn isolates, and followed by F. fujikuroi (27.7%), F. proliferatum (14.9%), F. subglutinans (7.1%), and F. concentricum (2.8%). A single mating type (MAT1-1) was found predominantly among the Gf complex isolates examined. Possible distinct subclades were detected within the populations of F. fujikuroi and F. proliferatum; however, this needs further confirmation. This is the first reported population-level characterization of putative fumonisin-producing Gf complex associated with rice and corn in Korea.