• Title/Summary/Keyword: Molecular Breeding

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Construction of a Genetic Linkage Map in Radish(Raphanus sativus L.) Using RAPD Markers (RAPD 마커를 이용한 무의 유전자지도 작성)

  • Ahn, Choon-Hee;Choi, Su-Ryun;Lim, Yong-Pyo;Chung, Hae-Joon;Yae, Byeong-Woo;Yoon, Wha-Mo
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.151-159
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    • 2002
  • Genetic map and molecular marker have a great importance in improving and facilitating crop breeding program as well as in genome analysis and map-based cloning of genes representing desirable characters. This study aimed at developing RAPD markers and constructing a genetic linkage map using 82 BC$_1$F$_1$individuals originated from the cross between '835' and B$_2$in radish (Raphanus sativus L.). One of the parents for genetic linkage map construction, '835'(P$_1$) of egg type is susceptible to Fusarium wilt and have medium resistance to virus infection and the other parent, B$_2$(P$_2$) of round type, is susceptible to Fusarium wilt and virus, Screening of 394 RAPD primers in BC$_1$F$_1$) population resulted in selecting 128 polymorphic markers which displayed 1:1 segregation pattern. Two markers failed to display 1:1 segregation and showed the segregation ratio skewed to maternal genotype. Selected markers were categorized into 14 linkage group based on LOD score represented by MAPMAKER/EXP program. Five groups composed of single marker among them were excluded from the linkage map, and consequently, the remaining groups are well matched with the number of radish chromosome (n=9). The linkage map constructed with 128 markers covers 1,688.3 cM and the average distance between markers was 13.8 cM. For developing STS marker, we determined the partial nucleotide sequence of OPE10 marker at both ends and designed a oligonucleotide primer pair based on this sequence. STS PCR using the primer pair displayed a single, clear band of which segregation is perfectly matched with that of OPE10 marker. This implies that RAPD markers could readily convert into clear and reliable STS markers.

Development of Efficient Screening Methods for Melon Plants Resistant to Fusarium oxysporum f. sp. melonis (멜론 덩굴쪼김병에 대한 효율적인 저항성 검정법 개발)

  • Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.70-82
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    • 2015
  • This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.

Efficient plant regeneration through callus induction from the hypocotyl of Perilla frutescens L var. Dayu ('다유들깨'품종의 하배축에서 캘러스를 통한 고효율 식물재분화)

  • Ruyue Xu;Ji-Hi Son;Hong-Gyu Kang;Hyeon-Jin Sun;Hyo-Yeon Lee
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.248-254
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    • 2023
  • This study was conducted to establish an efficient plant regeneration system in 'Dayu', a Korean variety of Perilla frutescens developed for seed oil production, in conjunction with the previously studied variety 'Namcheon'. The healthiest callus was formed on the hypocotyl explants cultured on a medium containing 0.1 mg/L NAA and 0.5 mg/L BA, outperforming the leaf and cotyledon samples. In both dark and long-day conditions, Dayu consistently exhibited significantly higher shoot regeneration rates compared with Namcheon. The highest shoot regeneration rates in Dayu were observed from the hypocotyl explants cultured on 0.1 mg/L NAA and 0.5 mg/L BA media, with shoot regeneration rates of 84.4% and 86.7% under dark and long-day conditions, respectively. Various combinations of plant growth regulators were tested to establish the optimal shoot regeneration conditions for Dayu hypocotyl explants. The results demonstrated that the highest shoot regeneration rate (90%) was achieved when 0.5 mg/L of BA was added to the medium without NAA. Among the regenerated shoots, 70.5% were normal plants, while 19.3% were abnormal. The addition of NAA or an increase in its concentration led to a higher occurrence of abnormal plants. After the regenerated shoots were transferred to 1/2 MS medium, roots were observed within 10-15 days. By day 30, they had developed into complete plants. The results obtained from the regeneration experiments with the perilla variety Dayu can valuably inform molecular breeding reliant on transformation techniques such as genome-editing and genetic modification technology.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Studies on the Characteristics of Volatile Fatty Acid Evolution from Fresh Animal Feces (축분의 휘발성 지방산 발현 양상 연구)

  • ;;;Hudson, Neale
    • Journal of Animal Environmental Science
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    • v.10 no.1
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    • pp.11-22
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    • 2004
  • This work was carried out to measure volatile fatty acids emissions from different manure (poultry, swine, cattle) incubated at $10^{\circ}C$, $25^{\circ}C$, and $37^{\circ}C$ for 6 days under anaerobic condition. Following are summary of these tests results. 1. Amounts of Acetic acid generated were 1,128.05mg/kg, 628.21mg/kg and 592.50mg/kg for swine, poultry, and cattle manure, respectively, during the period of incubation. In the case of swine and cattle manure, 83.87%(946.10mg/kg) and 57.49%(340.63mg/kg) from all the temperature treatments were produced in the $25^{\circ}C$, respectively. 83.57% in swine and 78.79% in cattle manure were intensively emerged from 3 day, 4 day and 5 day of the $25^{\circ}C$ treatment. In the case of poultry manure, 45.36%(284.93mg/kg) and 45.36%(284.93mg/kg) in the $25^{\circ}C$ and in the $37^{\circ}C$, respectively, were produced. Accordingly, acetic acid generated from poultry manure was characteristic of being mainly produced in more than $25^{\circ}C$. 2. Amounts of propionic acid generated were 238.56mg/kg, 162.14mg/kg and 155.49mg/kg for swine, poultry, and cattle manure, respectively, during the period of incubation. In the case of swine manure, 78.52%(187.32mg/kg) of propionate emitted from all the temperature treatments was produced in the $25^{\circ}C$ and 79.1% of them was intensively emerged from 3day, 4day and 5day of the $25^{\circ}C$ treatment. In the case of poultry manure, 35.12%(56.95mg/kg) and 45.89%(74.40mg/kg) of the propionate amounts were produced in the $25^{\circ}C$ and in the $37^{\circ}C$, respectively. In the case of cattle manure, 28.21% (43.86mg/kg) and 49.30% (76.66mg/kg) of the propionate amounts were produced in the $10^{\circ}C$ and in the $25^{\circ}C$, respectively. Accordingly, propionate evolved from poultry manure was characteristic of being mainly produced in more than $25^{\circ}C$ and from cattle manure, in less than $25^{\circ}C$, respectively. 3. Amount of butyric acid generated were 1,463.87mg/kg, 96.72mg/kg and 129.18mg/kg for swine, poultry, and cattle manure, respectively, during the period of incubation. The time intensively emerged from the period of incubation was differently generated from the incubation temperature and animal feces. 4. Amounts of iso-valeric acid generated were 6,885.99mg/kg, 399.28mg/kg and 307.47mg/kg for swine, cattle and poultry manure, respectively, during the period of incubation. In the case of swine and cattle manure, 28.22%(1,943.52mg/kg) and 48.56%(193.90mg/kg) in the $25^{\circ}C$, 68.76%(4,734.90mg/kg) and 46.93%(187.40mg/kg) in the $37^{\circ}C$, respectively, were occupied. Accordingly, iso-valeric acid evolved from swine and cattle manure was characteristic of being mainly produced in more than $25^{\circ}C$. In the case of poultry manure, 59.89%(184.13mg/kg) of iso-valeric acid generated from all the temperature treatments was produced in the $37^{\circ}C$ and 100% of them was intensively emerged from 2 day and 3 day of the $37^{\circ}C$ treatment.

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