Twenty Inter simple sequence repeat (ISSR) primers were used to assess genetic diversity of 64 Agaricus strainsincluding 45 A. bisporus strains and other 19 Agaricus spp. ISSR primers, (GA)T, (AG)YC, (GA)C and (CTC) amplified PCRpolymorphic bands between the Agaricus species or within A. bisporus strains. PCR polymorphic bands were inputted forUPGMA cluster analysis. The varieties, Saea, Saedo, Saejeong and Saeyeon that have recently been developed in Korea wereinvolved in the same group with closely genetic relationship of coefficient similarity over 0.92, whereas, other Korean strains weregenetically related to A. bisporus strains that were introduced from USA, Eroupe and Chinese. Furthermore, ISSR-PCRpolymorphism could potentially be used to identify homokaryon isolates.

우리나라와 전 세계의 여러 지역에서 수집한 비늘버섯속 18 균주와 개암비늘버섯 2 균주를 대상으로rDNA의 ITS region 염기서열과 genomic DNA의RAPD-PCR을 수행하였다. ITS1과 ITS2영역의 염기의수는 각각 233~271, 158~233 그리고 174~219 염기쌍으로 종에 따라 변이가 있었는데 ITS2영역의 염기서열이 ITS1의 영역보다 변이가 높았고 5.8S지역의 염기의 수는 비교적 변이가 적었다. 각각의 균주 간 유연관계를 알아보기 위해 ITS영역의 염기서열을 이용하여계통도를 작성한 결과 실험에 사용한 균주는 8개의 클러스터로 나누어지는 것으로 나타났으며 동일한 종의버섯은 동일한 클러스터에 속하는 것으로 나타났다.또한 20종류의 primer를 이용하여 비늘버섯속 버섯을대상으로 RAPD-PCR을 수행한 결과 15개의 primer가효과적으로 염색체 DNA를 증폭하는 것으로 나타났다.증폭의 양상은 primer의 종류와 종에 따라 변이가 있었다. 이 결과를 토대로 계통수를 작성한 결과 계통수는 ITS 영역의 PCR 결과와 매우 유사하였다. 본 실험결과, 실험에 사용한 비늘버섯속 버섯의 종과 계통 간의 유연관계는 높았으며, rDNA ITS 영역의 염기서열분석 결과를 이용해 공시된 각각의 비늘버섯 종을 분류하는데 유용하게 사용이 가능하였다.

[Aims] Lentinula edodes (Berk.) Pegler has many commercial strains both morphologically and physiologically similar to each other. At present, detection of polymorphism in rDNA-IGS region (Babasaki, 2006) and/or RAPD marker (Zhang and Molina, 1995) is generally used for strain typing of L. edodes. However, it is rather time-and-cost consuming. Inter-retrotransposon amplified polymorphism (IRAP)-PCR method mainly used for horticultural crops takes less time and lower in cost in strain typing (Kalendar et al, 1999). In this study, we designed IRAP primers for L. edodes and verified their strain typing efficiency. [Method] Thirty three strains were provided for this study. Either fungal cultures on PDA or fungal tissues of fruit bodies were cut into approximately 4 x 4 x 4 mm. Total DNA of each samples were extracted by DNeasy Plant Mini Kit (QIAGEN). For PCR, IRAP primer set and Pfu-X polymerase (greiner) were performed. Based on LTR (Long Terminal Repeat) sequence in L. edodes, we designed one set of primers amplifying the regions between retrotransposons. Ampricons were electrophored for 50 min at 100 V on 1.7 % agarose gel with GelRed (Biotium) and evaluated under UV irradiation. [Results] The products obtained by IRAP-PCR were determined using mini-gel electrophoresis system. The band patterns of IRAP-PCR products differ among strains except the ones having the same parental cultivar. The detected bands were bright and clear without smearing. The IRAP-PCR products of fungal cultures on PDA and correlating fungal tissues of fruit bodies showed the same band pattern, suggesting that the procedure is highly reproducible. Thus, it is considered that IRAP-PCR with short ranged (ca. 1 kb) electrophoresis is a time-efficient and practical strain typing method of L. edodes.

The diamondback moth, Plutella xylostella, is one of the most important pests of cole crops in the world and is the first insect to evolve resistance to Bt toxins in open-field populations. To search for useful molecular markers for Bt reistance monitoring, the PCR-based restriction fragment length polymorphism (RFLP) profiles of three aminopeptidase N (PxAPN1, PxAPN2 and PxAPN4) were determined for 15 representative regional field populations of P. xylostella. Most regional samples had similar RFLP patterns, whereas PxAPN1 from four regions and PxAPN4 from two regions showed different banding patterns after restriction enzyme treatment, but no differences were found in PxAPN2 among populations. The DNA sequence analysis revealed that a point mutation at the restriction site was responsible for the polymorphism of PxAPN1 but no mutations were observed in PxAPN4. Comparing amino acid sequences of PxAPNs from regional populations with reference PxAPNs (GenBank accession no. AAB70755) revealed that four regional populations possessed a point mutation in the Cry1A binding site of PxAPN1 and five regional populations possessed a deletion of eight amino acids in PxAPN4. These RFLP patterns were consistently observed in Southern regions of Korea, including Kyungsangnam-Do and Jeju-Do. The functional association of these RFLP with Bt resistance is currently under investigation

This research aimed to compare the detection methods of Anisakis simplex in Sea fish by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and macroscopic inspection. We examined 18 Trichiurus lepturus, 11 Scomber japonicus, and 65 Todarodes pacificus collected from the retail markets in the areas of Uljin, Kyuonggi province and Seoul. As the result of examinations, we found that detection rate of Anisakis simplex by macroscopic observation was 89% in Trichiurus lepturus, 90.9% in Scomber japonicus, 32.3% in Todarodes pacificus. The detection rate of Anisakis simplex by PCR-RFLP was 77.7% in Trichiurus lepturus, 81.8% in Scomber japonicus, 26.1% in Todarodes pacificus. We could conclude that PCR-RFLP method of Anisakis simplex was more specific rather than macroscopic observation.