• Title/Summary/Keyword: Pre-inoculation

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High-efficiency and Rapid Agrobacterium-mediated genetic transformation method using germinating rice seeds (벼 발아초기 종자를 이용한 고효율 단기형질전환 방법)

  • Lee, Hye-Jung;Abdula, Sailila E.;Jee, Moo-Geun;Jang, Dae-Won;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.251-257
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    • 2011
  • Rice is the most important crop as a model plant for functional genomics of monocotyledons. Rice is usually transformed using Agrobacterium tumefaciens. However, the transformation efficiency using previous method is still low. In this study, we established a new method by modifying the general Agrobacterium protocol especially in the inoculation and co-cultivation step. We directly inoculated Agrobacterium containing a CIPK15 gene under the control of CaMV 35S promoter and NOS terminator in the pCAM1300 vector into the pre-soaked seeds in N6D media for 24 hours. After 7 days of culture at $25^{\circ}C$, calli were formed on seeds cultured on the co-cultivation medium containing an antioxidant compound (1 mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (3 mg/L silver nitrate). We obtained 35 and 22 transgenic plants in rice cultivars, Gopumbyeo and Ilpumbyeo, with increase of transformation efficiency by 30.4% and 22.6%, respectively compared to the general transformation method. The new method in this study would lead to reduction of substantial labor and time to generate transgenic plants.

Effect of Bordeaux Mixture on Control of Rice Leaf Blast (벼 유기재배에서 석회보르도액을 이용한 벼 잎도열병 방제 효과)

  • Kang, Beom-Ryong;Kim, Seon-Gon;Kim, Do-Ik;Lee, Yong-Hwan;Choi, Kyong-Ju;Choi, Yong-Soo
    • Research in Plant Disease
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    • v.14 no.3
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    • pp.182-186
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    • 2008
  • Recently organic farming practice of rice has been emerged in Korea, but one of the major limiting factor is the no effective environmental-friendly agro-materials to control major plant diseases. Bordeaux mixture has been used effectively as a preventive agro-chemical. The aim of this study was to investigate efficacy of Bordeanux mixture on control of rice blast caused by Magnaporthe grisea which is one of the disruptive rice diseases in world-wide. In greenhouse experiment, pre-treatment of 6-6 type of Bordeaux mixture before inoculation of spore suspension of M. grasea showed 71 % of control value. In field experiment, preventive applications of 4-8 and 6-6 types of Bordeaux mixture showed over 71 % of the control value. Chemical injury on rice leaves were not found in the application concentrations of all types of Bordeaux mixture, but observed in applications of Bordeaux mixtures between 30 and 100 diluted concentrations. This results indicate Bordeaux mixture can be used as an effective environmental-friendly agro-chemical to control rice blast disease in the field.

The effect of post-incubation period and temperature treatment on the uniform primordia formation of Pleurotus ostreatus (느타리버섯 후배양 기간과 온도처리가 발이 균일도에 미치는 영향)

  • Jeoung, Yun-Kyeoung;Baek, Il-Sun;Kim, Jeong-Han;Chi, Jeong-Hyun;Lim, Jae Wook
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.223-227
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    • 2015
  • The study was conducted to investigate the effects of post-incubation period and temperature treatment conditions during incubation on the uniform primordia formation and cultural characteristics of oyster mushroom (Pleurotus ostreatus). Three kinds post-incubation period; 25, 30, 35 days and control were applied for 30 days while two kinds incubation room temperature $23^{\circ}C$ and $26^{\circ}C$ and control were used $20^{\circ}C$. The substrate temperature during pre-incubation was of 'Suhan No. 1' and 'Gonji No. 7'. Oyster mushroom varieties tended to increase between $24^{\circ}C$ to $26^{\circ}C$ at 11 to 15 days after inoculation and then they were maintained in treatment temperature during post -incubation period. The $CO_2$ occurrence was at the highest at 6,500 ppm for 'Suhan No. 1' and 5,800 ppm for 'Gonji No. 7' at the time of the highest temperature increase. The ratio of un-uniformal primordia formation and the ratio of non-commercial fruit body were reduced by 40%, 10.5%, respectively compared to control for 'Suhan No. 1' when in the post-incubation temperature was $26^{\circ}C$, and incubated for 10 days and 15 days treatment. Also, 'Gonji No. 7' was reduced by 19%, 9.5%, respectively when in the post-incubation temperature was $26^{\circ}C$, and incubated for 10 days treatment. Therefore, the higher post-incubation temperature of room and longer post-incubation period resulted in the higher percentage of primordia formation of two cultivars.

Isolation and Characterization of Plant Pathogen that Cause Soft Rot Disease in Napa Cabbage (배추무름병 원인균 분리 및 특성 연구)

  • Kwon, Young-Hee;Yoo, Ah-Young;Yu, Jong-Earn;Kang, Ho-Young
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1177-1182
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    • 2009
  • In order to establish in vitro infection model for research of plant pathogen based on tissue softening disease in napa cabbage, eighty independent bacterial strains were isolated from the softened napa cabbage tissues. Eight bacterial isolates were primarily screened with the generation of reproducible tissue softening disease to fresh napa cabbages within 24${\sim}$48 hours after inoculation. Through various microbiological biochemical and morphological examinations, three Gram (-) isolates which harbor independent biological properties were finally chosen, and named as RBI, RB2 and RB6. Collective results obtained from API 20E test and analyses of VITEK 2 COMPACT and nucleotide sequences of 165 rRNA of each isolate proposed that isolates RBI and RB2 are close to the Erwinia carotovora subsp. odorifera, and RB6 is close to the Erwinia carotovora subsp. carotovora. These isolates grew optimally at $30^{\circ}C$ with neutral pH culture condition. The isolates caused softening tissue disease with dose-dependent manner regardless of pre-surface damages of napa cabbage. Minimum dose to cause soft rot disease for RBI, RB2 or RB6 were $8.0{\times}10^8$ CFU/mt $10^9$ CFU/ml or $4.7{\times}10^6$ CFU/ml respectively. These isolates caused tissue softening disease to eggplant, paprika and napa cabbage out of 14 different tested vegetables, indicating that these isolates damages specific plant tissues. The bacterial isolates obtained in this research and in vitro plant infection model will be adapted in the understanding of the mechanism of pathogenesis by plant pathogen.

An Etiological Study on the Anthracnose Fungus of Pepper Caused by Colletotrichum dematium in Korea (고추에서 분리(分離)한 탄저병균(炭疽病菌) Colletotrichum dematium에 대한 병원학적(病原學的) 연구(硏究))

  • Chung, Bong-Koo;Chang, Sun-Hwa
    • The Korean Journal of Mycology
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    • v.12 no.4
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    • pp.153-157
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    • 1984
  • In order to investigate an etiological characteristics of the pepper anthracnose fungus infected with Collectotrichum dematium, this experiment was undertaken. Isolation percent of the fungus was 55.3% from the seed sample of Jewon and 0.3% from that of Eumsung Kun, according to seed health blotter method. Although the acervulus was similar to the known two Colletotricum species, C. acutatum and C. gloeosporioides, setae was not only straightly extended above the acervulus, but also shaped as falcate conidia ranging $12{\sim}31.2{\times}2.4{\sim}4.8{\mu}m$. Acervului of the fungus were evenly scattered on the surface of pepper seed coat, and caused seedling blight after seed germination. Pre-and post-emergence seedling blight as well as foliar lesion and fruit rot was confirmed by inoculating conidial suspension. No typical anthracnose sysmptom was obtained from soybean seedlings, welsch onion and spinach seedlings upon inoculation. Therefore, this fungus is said to the undescribed form species of the fungus of pepper in Korea (Colletotricum dematium f. sp. capsicum).

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Evaluation of Streptomyces padanus IA70-5 Strain to Control Hot Pepper Anthracnose (Colletotrichum acutatum) (고추 탄저병 (Colletotrichum acutatum) 방제를 위한 Streptomyces padanus IA70-5의 평가)

  • Chi, Tran Thi Phuong;Choi, Okhee;Kwak, Youn-Sig;Son, Daeyoung;Lee, Jeung Joo;Kim, Jinwoo
    • Journal of agriculture & life science
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    • v.46 no.3
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    • pp.37-45
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    • 2012
  • To select bacterial strains with antifungal activity against an anthracnose fungal disease causing damage severely on hot pepper, previous isolates obtained from plant root samples were screened. Among 457 isolates, IA70-5 isolate was finally selected and identified as Streptomyces padanus based on 16S rDNA sequence analysis. Strain IA70-5 is non-pigmenteous, non-mobile, and filamentous. S. padanus IA70-5 inhibited effectively the mycelium growth, spore germination, and appressorium formation of Colletotrichum acutatum in vitro. The results of this study demonstrated that IA70-5 strain, especially applied on fruit of hot pepper, decreased disease incidence 90% for pre-inoculation before pathogen treatment. Taken together, S. padanus IA70-5 strain is a promising biological control agent to control of a major fungal pepper disease, anthracnose.

Disease Resistance-Based Management of Alternaria Black Spot in Cruciferous Crops (병 저항성 기반 십자화과 작물의 검은무늬병 관리)

  • Young Hee Lee;Su Min Kim;Seoung Bin Lee;Sang Hee Kim;Byung-Wook Yun;Jeum Kyu Hong
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.363-376
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    • 2023
  • Alternaria black spots or blights in cruciferous crops have been devastating diseases worldwide and led to economic losses in broccoli, Chinese cabbage, kale, radish, rapeseed, etc. These diseases are caused by different Alternaria spp., including A. brassicae, A. brassicicola and A. raphani transmitted from infected seeds or insect vectors. Efforts to excavate disease resistance traits of cruciferous crops against Alternaria black spots or blights have been demonstrated. Genetic resource of disease resistance was investigated in the wild relatives of cruciferous crops, and different cultivars were screened under different inoculation conditions. Development of the disease-resistant lines against Alternaria black spots or blights was also tried via genetic transformation of the cruciferous crops using diverse plant defence-associated genes. Plant immunity activated by pre-treatment with chemicals, i. e. β-amino-n-butyric acid and melatonin, was suggested for reducing Alternaria black spots or blights in cruciferous crops. The disease resistance traits have also been evaluated in model plant Arabidopsis originating from different habitats. Various plant immunity-related mutants showing different disease responses from wild-type Arabidopsis provided valuable information for managing Alternaria black spots or blights in cruciferous crops. In particular, redox regulation and antioxidant responses altered in the Alternaria-infected mutants were discussed in this review.

Attachment of Pasteuria penetrans Endospores to Meloidogyne spp. Juveniles Affected by Temperatures and the Nematode species (선충기생세균(Pasteuria penetrans) 내생포자의 뿌리혹선충(Meloidogyne spp.) 유충 부착에 대한 온도와 선충종의 영향)

  • Cho, Myoung Rae;Kim, Hyung Hwan;Kang, Taek Joon;Ahn, Seung Joon;Yang, Chang Yul;Jeon, Sung Wook
    • Korean journal of applied entomology
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    • v.52 no.2
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    • pp.109-113
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    • 2013
  • A greenhouse soil infested with an obligate nematode parasitic bacterium, Pasteuria penetrans, was used to test the effect of temperatures on the endospore attachment to root-knot nematode, Meloidogyne arenaria, juveniles (J2). Freshly hatched J2s were inoculated to the soil in petri dish and incubated under different temperatures of $20^{\circ}C$, $25^{\circ}C$, $30^{\circ}C$, and $35^{\circ}C$ for 7 days. The endospore attachment rates were 100% in all the temperatures, while the number of endospores attached per J2 was highest in $25^{\circ}C$ with 28.3 endospores/J2 followed by 20.2, 18.6, and 13.6 in $30^{\circ}C$, $20^{\circ}C$, and $35^{\circ}C$, respectively. When the soil was pre-treated under different temperatures before the J2 inoculation, the endospore attachment rates significantly decreased from 60% in room temperature to $25^{\circ}C$, 31.7, 8.3, 5.0, and 0% after the soil incubation in $-30^{\circ}C$, $4^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $100^{\circ}C$ for 10 days, respectively. The endospore numbers attached per J2 were 3.5, 4.3, 1, 1, and 0 when the soil was pre-treated in $-30^{\circ}C$, $4^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $100^{\circ}C$, respectively, which were lower than 5.3/J2 of room temperature treated soil. The P. penetrans isolate in the soil showed nematode species-specific endospore attachment characteristics with 100% attachment rate only on M. arenaria J2s while the rates were 0% on M. hapla and M. incognita J2s.

Quality Preservation of Shredded Carrots Stored in UV LED Packaging System (자외선 LED 포장용기 시스템에 의한 포장절단당근의 품질보존)

  • Kim, Nam Yong;Lee, Dong Sun;An, Duck Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.135-140
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    • 2014
  • Pre-storage ultra-violet (UV) light treatment on fresh produce is known to inactivate the contaminated microorganisms, activate the defense system, and delay ripening extending the shelf life. As UV light emitting diode (LED) becomes available at a relatively low price, continuous or intermittent UV treatment during chilled storage is possible in a container or package. This study attempted an in situ UV LED treatment on fresh produce stored under a refrigerated container in order to see its potential in the fresh produce storage and further optimize its application conditions. The effect of in-container UV LED irradiation on the quality preservation of shredded carrots was investigated in the air and modified atmosphere (MA) conditions. Two sets of experiment with Escherichia coli inoculation and with natural microbial flora in the air (two 30 minute on-off cycles of 1 $diode/dm^2$ per day at a location above 2 cm) showed a clear and significant effect of the UV LED irradiation on the suppression of microbial growth: 280 nm was the most effective by maintaining a lower microbial count by at least 0.5 log (CFU/g) throughout the 6 day storage period. The carotenoids content of shredded carrots subjected to UV LED treatment at 365 and 405 nm in the air was higher than that of the control shredded carrots. In MA condition of $O_2$ of 1.2~4.3% and $CO_2$ of 8.4~10.6% being indifferent with LED wavelengths, 280 nm UV LED irradiation was also effective in inhibiting the microbial growth. While there was no observed difference in the carotenoids content between untreated and UV LED-treated shredded carrots in MA, UV LED irradiation at 365 and 405 nm was slightly better in DPPH radical scavenging activity. The use of UV LED in storage container or package seems to give the benefits of preserving the microbial and nutritional qualities of minimally processed fruits and vegetables.

Simple Method to Discriminate the Fungicide Resistant Botrytis cinerea Strain in Tomatoes (토마토 잿빛곰팡이병균 약제저항성 간이 판별법)

  • Lee, Mun Haeng;Lee, Hee Kyoung;Kim, Sung Eun;Lee, Hwan Gu;Lee, Sun Gye;Yu, Seung Hun;Kim, Young Shik;Kim, Sang Woo;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.41 no.3
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    • pp.172-180
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    • 2013
  • Grey mold infection rate in tomato was investigated with the inoculation of dead flowers on Botrytis selective media. The grey mold infection rate of flower after fruiting were higher in the order of after 45 days, after 25 days, and fruiting day with 100%, 87% and 65%, respectively. The number of infected flowers were increased with time increase after the flowering before fruiting. BSM (Botrytis selective medium) was used to check grey mold infection rate depending on the flowering stage and cultivar. Grey mold infection rate depending on the flowering stage was similar in all the beef-tomato cultivar as 1.5~5% at preflowering, 1.5~45% at flowering and 75~90% at fruiting. On the other hand, cherry tomato cultivar "KoKo" had lower infection rates of 0~3.5% at pre-flowering, 10~30% at flowering and 20~50% at fruiting. These resulted from the fact that beaf-tomato cultivar have much bigger flowers and larger amount of pollens compared to those of cherry tomato cultivar. The amounts of falling pollens of Botrytis spp. were checked for beaf-tomato cultivar and cherry tomato cultivar using BSTM. The amounts of falling pollens were increased as growth period was extended, and the amount of spores increased rapidly during the outbreak of grey mold. Twelve field trials in Buyeo and Iksan areas showed that Fluazinam, and Diethofencarb+Carbendazim were effective fungicides to control tomato grey mold, and these results were similar to those of field trials with BSTM. This is the first report of Fluazinam as a effective fungicide for the control of grey mold of tomato even though it has not been registered yet for the control of gray mold in tomato.