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Specific Primer for Detection of Jujube Witches' Broom Phytoplasma Group (16SrV) in Korea
Sangsub Han 한국식물병리학회 2005 Plant Pathology Journal Vol.21 No.1
In order to diagnose and differentiate jujube witches'broom (JWB) phytoplasma rapidly, oligonucleotide primer pair, 16Sr(V) F/R, for polymerase chain reactions (PCRs) was designed on the basis of 16S rRNA sequences of JWB phytoplasma. The PCR employing phytoplasma universal primer pair P1/P7 consistently amplified DNA in all tested phytoplasma isolates. But no phytoplasma DNA was detected from healthy jujube seedlings. The nested PCR, the primer pair 16S(V) F/R,about 460 bp fragment, amplified DNA in all tested JWB and related phytoplasmas including ligustrum witches’ broom phytoplasma of the 16S rRNA group V,but no DNA amplification was detected from other phytoplasma strains such as groups 16SrI (Aster yellows) and 16SrXII (Stolbur group) in which mulberry dwarf phytoplasma and chrysanthemum witches’ broom phytoplasma belong to, respectively.The same results were obtained from both Korean and Chinese isolates of JWB phytoplasma. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16SrV group-specific primer pair 16S(V) F/R could detect group V phytoplasmas rapidly and easily, in particular JWB phytoplasma.
Balclutha punctata 에 의한 뽕나무 오갈병 파이토플라스마의 전반
한상섭 ( Sangsub Han ) 한국산림과학회 2012 한국산림과학회지 Vol.101 No.4
The present study was initiated to find out the insect vector of mulberry dwarf phytoplasma. Among the collected insects from mulberry cultivation region the sucking insects (Hishimonas sellatus Bothrogonia japonica and Balelutha punctata) were selected for transmission of mulberry dwarf phytoplasma. Sucking insects H.sellatus and B. punctata successfully transmitted phytoplasma to healthy seedling mulberry and periwinkle plants by inoculation-feeding. Typical dwarf symptom was observed in mulberry tree but very slender twigs on stem and formatted small leaves were found on periwinkle plants which are typical witches` broom symptoms. The phytoplasmas were detected in all samples exception sucking insects B. japonica using phytoplasma universal primer pair R16F2n/R2.
Cha, Byeongjin,Han, Sangsub The Korean Society of Plant Pathology 2002 Plant Pathology Journal Vol.18 No.6
Heteroduplex mobility assay (HMA) and single-strand conformation polymorphism (SSCP) analyses combined with PCR were developed for genetic differentiation of various phytoplasma isolates. In the HMA and SSCP analyses, differences in the mobility shifts and the SSCP band patterns identified three distinct types of phyto-plasmas: Type Ⅰ, jujube witches'-broom (JWB) and ligustrum witches'-broom (LiWB); Type Ⅱ, mulberry dwarf(MD) and sumac witches'-broom (SuWB); and Type Ⅲ, paulownia witches'-broom (PaWB). Results of the sequence analyses revealed that phytoplasmas of JWB and MD had 100% homology with LiWB and SuWB, respectively. On the other hand, PaWB phyto-plasma had 97.8% homology with MD phytoplasma. The PCR-HMA and SSCP techniques were very useful in determining variations in sequence among several isolates of phytoplasmas. Furthermore, the methods were rapid, economical, highly sensitive, and easy to handle with the gels.
Cha, Byeongjin,Han, Sangsub The Korean Society of Plant Pathology 2002 Plant Pathology Journal Vol.18 No.2
Using phytoplasma universal primer pair Pl and P7, a fragment of about 1.8 kb nucleotide sequences of 16S rRNA gene and 16S-23S rRNA intergenic spacer region, and a portion of 23S rRNA gene of jujube witches'broom (JWB) and mulberry dwarf(MD) phytoplasmas were determined. The nucleotide sequences of JWB and MD were 1,850 bp and 1,831 bp long, respectively. The JWB phytoplasma sequence was aligned with the homologous sequence of MD phytoplasma. Twenty-eight base insertions and nine base deletions were found in the JWB phytoplasma sequence compared with that of MD phytoplasma. The similarity of the aligned sequences of JWB and MD was 84.8%. The near-complete 16S rRNA gene DNA sequences of JWB and MD were 1,529 bp and 1,530 bp in length, respectively, and revealed 89.0% homology. The 16S-23S rRNA intergenic spacer region DNA sequences were 263 bp and 243 bp in lengths respectively, while homology was only 70% and the conserved tRNA-lle gene of JWB and MD was located into the intergenic space region between 16S-23S rRNA gene. The nucleotide sequences were 77 bp long in both JWB and MD, and showed 97.4% sequence homology. Based on the phylogenetic analysis of the two phytoplasmas, the JWB phytoplasma belongs to the Elm yellow phytoplasma group (16S rV), whereas, the MD phytoplasma belongs to the Aster yellow group (16S rI).
장동헌 ( Dongheon Jang ),이상현 ( Sanghyun Lee ),한상섭 ( Sangsub Han ) 한국협동조합학회 2010 韓國協同組合硏究 Vol.28 No.1
In this study, we examined the development plans of producer organization on the basis of the investigation on producers`` awareness and distribution status of rhododendron farms. The key contents and suggestions are as follows. First, the technical standard of producers was of a high level and the difficulty experienced by producers was market and price instability. In addition, soil management was limited to fertilization and control of insect pests. Also, there was insufficiency of discussions on farming plans among producers. Therefore, it is necessary to establish composite farming plans as well as to pursue joint purchasing of farming materials, scientific soil management and introduction of programs toactivate the organization. Second, rhododendron distribution mainly took place as trading at farming yards and the price was determined largely by merchants. For marketing, there was lack of distribution information and insufficient securing of trading partners. Therefore, it is necessary to improve price negotiation power, to establish a landscaping plant distribution center and to promote the 6th industrialization, etc. Third, in the long-term perspective, it will be necessary to establish the basis for R&D and implementation of hub for rhododendron industry.