• 제목/요약/키워드: LATE-PCR

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Epidemiology and Control of Strawberry Bacterial Angular Leaf Spot Disease Caused by Xanthomonas fragariae

  • Kim, Da-Ran;Gang, Gun-hye;Jeon, Chang-Wook;Kang, Nam Jun;Lee, Sang-woo;Kwak, Youn-Sig
    • The Plant Pathology Journal
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    • 제32권4호
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    • pp.290-299
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    • 2016
  • Strawberry bacterial angular leaf spot (ALS) disease, caused by Xanthomonas fragariae has become increasingly problematic in the strawberry agro-industry. ALS causes small angular water-soaked lesions to develop on the abaxial leaf surface. Studies reported optimum temperature conditions for X. fragariae are $20^{\circ}C$ and the pathogen suffers mortality above $32^{\circ}C$. However, at the nursery stage, disease symptoms have been observed under high temperature conditions. In the present study, results showed X. fragariae transmission was via infected maternal plants, precipitation, and sprinkler irrigation systems. Systemic infections were detected using X. fragariae specific primers 245A/B and 295A/B, where 300-bp and 615-bp were respectively amplified. During the nursery stage (from May to August), the pathogen was PCR detected only in maternal plants, but not in soil or irrigation water through the nursery stage. During the cultivation period, from September to March, the pathogen was detected in maternal plants, progeny, and soil, but not in water. Additionally, un-infected plants, when planted with infected plants were positive for X. fragariae via PCR at the late cultivation stage. Chemical control for X. fragariae with oxolinic acid showed 87% control effects against the disease during the nursery period, in contrast to validamycin-A, which exhibited increased efficacy against the disease during the cultivation stage (control effect 95%). To our knowledge, this is the first epidemiological study of X. fragariae in Korean strawberry fields.

Stem Rot on Adzuki Bean (Vigna angularis) Caused by Rhizoctonia solani AG 4 HGI in China

  • Sun, Suli;Xia, Changjian;Zhang, Jiqing;Duan, Canxing;Wang, Xiaoming;Wu, Xiaofei;Lee, Suk-Ha;Zhu, Zhendong
    • The Plant Pathology Journal
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    • 제31권1호
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    • pp.67-71
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    • 2015
  • During late August and early September 2011, stem rot symptoms were observed on adzuki bean plants (Vigna angularis) growing in fields located in Beijing and Hebei Province, China, respectively. In this study, four isolates were obtained from infected stems of adzuki bean plants. Based on their morphology, and sequence and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses of the ribosomal DNA internal transcribed spacers (rDNA-ITS) region, the four isolates were identified as Rhizoctonia solani in anastomosis group (AG) 4 HGI. Pathogenicity tests showed that all isolates were strongly pathogenic to adzuki bean and resulted in serious wilt symptoms which was similar to observations in the fields. Additionally, the isolates infected several other crops and induced related rot on the roots and basal stems. To our knowledge, this is the first report of Rhizoctonia solani AG 4 HGI causing stem rot on adzuki bean.

cDNA cloning and expression pattern of Cinnamate-4-Hydroxylase in the Korean black raspberry

  • Baek, Myung-Hwa;Chung, Byung-Yeoup;Kim, Jin-Hong;Kim, Jae-Sung;Lee, Seung-Sik;An, Byung-Chull;Lee, In-Jung;Kim, Tae-Hoon
    • BMB Reports
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    • 제41권7호
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    • pp.529-536
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    • 2008
  • Cinnamate-4-hydroxylase (C4H) is a key enzyme in the phenylpropanoid pathway, which is responsible for synthesizing a variety of secondary metabolites that participate in development and adaptation. In this study, we isolated a full-length cDNA of the C4H gene from the Korean black raspberry (Rubus sp.) and found that this gene existed as a single gene. By comparing the deduced amino acid sequence of Rubus sp. C4H with other sequences reported previously we determined that this sequence was highly conserved among widely divergent plant species. In addition, quantitative real time PCR studies indicated that the C4H gene had a differential expression pattern during fruit development, where gene expression was first detected in green fruit and was then remarkably reduced in yellow fruit, followed by an increase in red and black fruit. To investigate the two peaks in expression observed during fruit development and ripening, we measured the flavonoid content. The content of the major flavanol of Korean black raspberry fruits was determined to be highest at the beginning of fruit development, followed by a gradually decrease according to the developmental stages. In contrast, the content of anthocyanins during the progress of ripening was dramatically increased. Our results suggest that the C4H gene in Korean black raspberry plays a role during color development at the late stages of fruit ripening, whereas the expression of C4H gene during the early stages may be related to the accumulation of flavanols.

Development of Neutaribeosut Varieties (Pleurotus ostreatus) Chongpung, Myongwol

  • Lee, He-Duck;Kim, Hong-Kyu;Kim, Young-Gyun;Han, Kyu-Heng;Min, Kwang-Ho;Jeong, Tae-Wo;Lee, Jae-Jung;Jeon, Tae-Sun;Kim, Duck-wan;Kim, Hun-Sick;Han, Jong-Ku
    • Plant Resources
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    • 제3권2호
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    • pp.105-109
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    • 2000
  • This study was carried out to find a useful mushroom at Chungnam Agricultural Research And Extention Service. Twenty materials used were collected from domestic and exotic area. These races were compared bontanical characteristics to leading varieties by PCR-RAPD methods. Mycelial growth temperature of Chongpung and Myongwol were at 20 to 25 $^{\circ}C$ and 25 to 30 $^{\circ}C$ at PDA medium, respectively mycelial growth of these varieties were similiar at pH 6.5 to 7.5. In case of mushroom cultivation temperature ranges, Chongpung was at 5 to 26$^{\circ}C$ and Myongwol was at 7 to 28$^{\circ}C$, but the optimum temperature range of these were appeared at 15 to 19$^{\circ}C$. Culture temperature of these was 23$^{\circ}C$ and period of mycelial culture was needed 23 to 24 days under 850cc/pp, while was needed 11 to 12 days at waste cotton medium. Cap color of these at first inducing mushroom was all dark blue, but at late growing stages Chongpung was shown as grey, and Myongwol was shown as dark grey. Yield of Chongpung was appeared as 46kg/3.3$m^2$ and that of Myongwol was 41kg /3.3$m^2$, while Chunchu No2 as check was 40kg/3.3$m^2$. Results from PDA medium and PCR-RAPD analysis two of these were different from others.

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Dynamics of Bacterial Communities of Lamb Meat Packaged in Air and Vacuum Pouch during Chilled Storage

  • Wang, Taojun;Guo, Huiyuan;Zhang, Hao;Ren, Fazheng;Zhang, Ming;Ge, Shaoyang;Luo, Hailing;Zhao, Liang
    • 한국축산식품학회지
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    • 제39권2호
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    • pp.209-221
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    • 2019
  • In this study, the changes in microbial communities of lamb meat packaged in the air (plastic tray, PT) and in a vacuum pouch (VAC) were assessed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) during the storage at $4^{\circ}C$. For the PT lamb, the total viable count (TVC) was $10^7CFU/g$ on Day 5, and the dominated bacteria were Pseudomonas fragi, P. fluorescens, and Acinetobacter spp. For the VAC lamb, the TVC was $10^7CFU/g$ on Day 9, and the dominated bacteria were lactic acid bacteria, including Carnobacterium divergens, C. maltaromaticum, and Lactococcus piscium. One strain of Pseudomonas spp. also appeared in VAC lamb. The relative abundance of Enterobacteriaceae in VAC lamb was higher than that PT lamb, indicating a more important role of Enterobacteriaceae in spoilage for VAC lamb than that of PT lamb. The microbial compositions changed faster in the lamb stored in a PT than that stored in a VAC, and microbial community compositions of the late storage period were largely different from those of the early storage period for both the conditions. The findings of this study may guide improve the lamb hygiene and prolong the shelf life of the lamb.

The spatio-temporal expression analysis of parathyroid hormone like hormone gene provides a new insight for bone growth of the antler tip tissue in sika deer

  • Haihua Xing;Ruobing Han;Qianghui Wang;Zihui Sun;Heping Li
    • Animal Bioscience
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    • 제37권8호
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    • pp.1367-1376
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    • 2024
  • Objective: Parathyroid hormone like hormone (PTHLH), as an essential factor for bone growth, is involved in a variety of physiological processes. The aim of this study was to explore the role of PTHLH gene in the growth of antlers. Methods: The coding sequence (CDS) of PTHLH gene cDNA was obtained by cloning in sika deer (Cervus nippon), and the bioinformatics was analyzed. The quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the differences expression of PTHLH mRNA in different tissues of the antler tip at different growth periods (early period, EP; middle period, MP; late period, LP). Results: The CDS of PTHLH gene was 534 bp in length and encoded 177 amino acids. Predictive analysis results revealed that the PTHLH protein was a hydrophilic protein without transmembrane structure, with its secondary structure consisting mainly of random coil. The PTHLH protein of sika deer had the identity of 98.31%, 96.82%, 96.05%, and 94.92% with Cervus canadensis, Bos mutus, Oryx dammah and Budorcas taxicolor, which were highly conserved among the artiodactyls. The qRT-PCR results showed that PTHLH mRNA had a unique spatio-temporal expression pattern in antlers. In the dermis, precartilage, and cartilage tissues, the expression of PTHLH mRNA was extremely significantly higher in MP than in EP, LP (p<0.01). In the mesenchyme tissue, the expression of PTHLH mRNA in MP was significantly higher than that of EP (p<0.05), but extremely significantly lower than that of LP (p<0.01). The expression of PTHLH mRNA in antler tip tissues at all growth periods had approximately the same trend, that is, from distal to basal, it was first downregulated from the dermis to the mesenchyme and then continuously up-regulated to the cartilage tissue. Conclusion: PTHLH gene may promote the rapid growth of antler mainly through its extensive regulatory effect on the antler tip tissue.

치주인대섬유모세포의 분화과정에서 아미노산 수송계 L의 발현 (Expression of amino acid transport system L in the differentiation of periodontal ligament fibroblast cells)

  • 황규영;김도경;김창현;장현선;박주철;최성미;김병옥
    • Journal of Periodontal and Implant Science
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    • 제36권3호
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    • pp.783-796
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    • 2006
  • The periodontium is a topographically complex organ consisting of epithelial tissue, soft and mineralized tissues. Structures comprising the periodontium include the gingiva, periodontal ligament (PDL) , cementum and the alveolar bone. The molecular mechanism of differentiation in PDL fibroblast cells remain unclear. Amino acid transporters play an important role in supplying nutrition to normal and cancer cells and for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the Na+-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In this study, the expression pattern of amino acid transport system L was, therefore, investigated in the differentiation of PDL fibroblast cells. To determine the expression level of amino acid transport system L participating in intracellular transport of amino acids in the differentiation of PDL fibroblast cells, it was examined by RT-PCR, observation of cell morphology, Alizaline red-S staining and uptake analysis after inducing experimental differentiation in PDL fibroblast cells isolated from mouse molar teeth. The results are as follows. 1. The LAT1 mRNA was expressed in the early stage of PDL fibroblast cell differentiation. This expression level was gradually reduced by differentiation- inducing time and it was not observed after the late stage. 2. The expression level of LAT2 mRNA was increased in time-dependent manner during differentiation induction of PDL fibroblast cells. 3. There was no changes in. the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during differentiation of PDL fibroblast cells. 4. The expression level of ALP mRNA was gradually increased and the expression level of Col I mRNA was decreased during differentiation of PDL fibroblast cells. 5. The L-leucine transport was reduced by time from the early stage to the late stage in PDL fibroblast cell differentiation. As the results, it is considered that among neutral ammo acid transport system L in differentiation of PDL fibroblast cells, the LATl has a key role in cell proliferation in the early stage of cell differentiation and the LAT2 has an important role in the late stage of cell differentiation for providing cells with neutral amino acids including several essential amino acids.

Potato virus Y에 의한 하령 감자의 괴경 괴저증상 (Superficial Tuber Necrosis in Potato Cultivar 'Haryeong' Caused by Potato virus Y)

  • 이영규;김점순;김주일;박영은
    • 식물병연구
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    • 제19권2호
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    • pp.90-94
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    • 2013
  • 하령 감자는 전분함량이 높고, 역병에 저항성이며 맛이 좋은 품종으로 2005년도에 신품종으로 등록되었다. 2010년 저장 중인 하령 씨감자에서 심한 표피의 괴저와 표면이 융기되고 원형의 괴저 병반이 생기는 괴경 괴저 증상이 발생하였다. 괴경 괴저 증상의 감자를 PVY 진단용 프라이머를 이용한 RT-PCR 분석 결과 모두 PVY가 검출되었다. 괴저증상 감자에서 검출된 $PVY^{Hkr}$ 외피단백질의 유전자 염기서열을 분석하였고 $PVY^{Kor}$, $PVY^N$, $PVY^{NTN}$, $PVY^O$, $PVY^C$ 계통과 상동성을 비교한 결과 $PVY^{Hkr}$은 2005년에 보고된 $PVY^{Kor}$와 2개의 염기를 제외하고 정확히 일치하였다. PVY 감염이 저장 중인 하령 품종에서 같은 병징을 일으키는지 확인하기 위해 5품종의 감자(하령, 수미, 대서, 대지, 추백)와 2종의 바이러스(PVY, PLRV)를 이용하여 생물검정을 실시하였다. 그 결과, 괴경 괴저 증상은 PVY에 감염된 저장 중인 하령 품종에서만 나타났다.

MicroRNA-101 Inhibits Cell Proliferation, Invasion, and Promotes Apoptosis by Regulating Cyclooxygenase-2 in Hela Cervical Carcinoma Cells

  • Huang, Fei;Lin, Chen;Shi, Yong-Hua;Kuerban, Gulinar
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권10호
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    • pp.5915-5920
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    • 2013
  • Aim: Although aberrant miRNA expression has been documented, altered miR-101 expression in cervical cancer and its carcinogenic effects and mechanisms remain unexplored. The aim of our study was to investigate the role of miR-101 alteration in cervical carcinogenesis. Methods: Expression of miR-101 was examined by quantitative real-time reverse transcriptase PCR (qRT-PCR) in Hela cells. After modulating miR-101 expression using miR-101 mimics, cell growth, apoptosis and proliferation, and migration were tested separately by MTT or flow cytometry and cell wound healing assay and protein expression was detected by qRT-PCR. The expression of COX-2 in Hela cell was also examined by immunohistochemical staining and the correlation with miR-101 expression was analysed. Results: The miR-101 demonstrated significantly low expression in Hela cell. When we transfected miR-101 mimics into Hela cells, the modulation of miR-101 expression remarkably influenced cell proliferation, cycling and apoptosis: 1) The expression of microRNA-101 tended to increase after transfection; 2) Overexpression of miR-101 was able to promote cell apoptosis, the apoptosis rate being markedly higher (97.6%) than that seen pre-transfection (12.2%) (P<0.05); 3) The miR-101 negatively regulates cell migration and invasion, scratch results being lower ($42.7um{\pm}2um$) than that observed pre-transfection ($181.4um{\pm}2um$); 4) miRNA-101 inhibits the proliferation of Hela cells as well as the level of COX-2 protein, which was negatively correlated with miR-101 expression. Conclusions: Overexpression of miR-101 has obvious inhibitory effects on cell proliferation, migration and invasion. Thus reduced miR-101 expression could participate in the development of cervical cancer at least partly through loss of inhibition of target gene COX-2, which probably occurs in a relative late phase of carcinogenesis. Our data suggest an important role of miR-101 in the molecular etiology of cancer and indicate potential application of miR-101 in cancer therapy.

자궁경부암세포에서 방사선조사시 차등 발현되는 유전자 동정 (Identification of Differentially Expressed Radiation-induced Genes in Cervix Carcinoma Cells Using Suppression Subtractive Hybridization)

  • 김준상;이영숙;이증훈;이웅희;성은영;조문준
    • Radiation Oncology Journal
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    • 제23권1호
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    • pp.43-50
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    • 2005
  • 목적 : 자경경부암세포에서 polymeric chain reaction (PCR)원리를 이용한 suppression subtractive hybridization (SSH) 방법으로 방사선조사 시 차등 발현되는 유전자를 동정하고자 하였다. 대상 및 방법 : 자궁경부암세포주인 HeLa세포주에 방사선조사 전과 후 총 RNA와 poly $(A)^+$ mRNA를 분리였다. SSH방법으로 forward 및 reverse-subtracted cDNA libraries를 만들었다. 차등 발현된 유전자를 screening하기 위해 reverse Northern blotting (dot blot analysis)을 이용하여 각각의 library에서 88개의 클론을 선택하였고 Nothern blotting으로 확인 후 sequencing하였다. 결과 : screening상 176개 클론 중 forward-subtracted library에서 10개의 유전자가 reverse-subtracted library에서 9개의 유전자가 동정되었다. forward-subtracted library로부터 3개의 유전자가 Northern blotting에 의하여 확인되었고 이중 telomerase catalytic subunit and sodium channel-like protein 유전자와 1개의 ESTs (expressed sequence tags) 유전자가 방사선선량에 따라 증가하쳐다. 결론 : 본 연구를 통해 자궁경부암세포주에서 방사선에 의해 유도되는 유전자를 SSH 방법을 통해 동정할 수 있었다. 그러나 이러한 유전자가 어떤 생물학적인 기능을 갖고 있는지에 대한 계속적인 연구가 필요하다