We quantified temporal and spatial changes in the habitat for fish populations, the distribution of mandarin fish(Siniperca scherzeri) and an introduced species, largemouth bass(Micropterus salmoides) in Lake Paro and inflowing streams. The number of fish species identified in Lake Paro and the tributary streams included 10 families, 24 species and 10 families 30 species, respectively. The dominant fish species in Lake Paro were Zacco platypus, Hemibarbus labeo, Squalidus gracilis majimae, S. scherzeri and Tridentiger brevispinis, Z. platypus, Z. koreanus, and S. gracilis majimae in the inflowing streams. Although the habitat segregation for S. scherzeri and M. salmoides occurs, these two species showed the use of the fishes of the family Gobiidae as an important prey item based on IRI analysis. S. scherzeri and M. salmoides preyed mainly on T. brevispinis(67.4 %) and R. brunneus(84.0 %), respectively. The species preyed on by S. scherzeri and M. salmoides were benthic fishes that inhabit shallow water depths around the lake and have little swimming ability.

본 연구는 2011년∼2013년 사이에 춘천시에 위치하고 종적으로 분포하는 2개 하천(공지천, 신촌천)을 대상으로 각각 2개의 조사 지점에서 저서성 대형무척추동물 군집의 계절 및 공간별 조사를 시행하였다. 본 연구는 하천에서의 인위적인 교란이 저서성 대형무척추동물의 종수 및 개체수의 감소에 미치는 영향을 고찰하였다. 본 연구의 상류구간인 신촌천(St.1, St.2)과는 달리 공지천 구간(St.3, St.4)은 공지천생태하천조성사업의 진행에 따른 인위적인 교란에 빈번하게 노출된 구간이었다. 이 사업 이후 물 흐름의 정체시간이 장기화됨으로 인하여 2012년 하절기에 신촌천 상류(St.1)와 공지천 하류(St.4) 사이에 커다란 수온의 차이(최대 9℃까지) 및 30℃ 이상 수온이 증가하는 현상을 보였다. 또한 매년 몬순강우는 저서성 대형무척추동물의 종수 및 개체수 그리고 종 다양도지수의 감소를 초래하였다. 분류군에 따른 우점 양상을 보면, 신촌천 상류에서만 날도래류가 우점하였을 뿐, 그 외의 지점들에서는 파리류가 우세하게 출현하였다. 저서성 대형무척추동물의 시·공간적인 변동은 인위적인 하상의 교란(하상변화, 수온 증가) 및 몬순강우 등으로 인한 토사유입과 같은 물리적인 요인들의 영향을 반영하는 지표생물로서 중요하다.

Citrus canker caused by Xanthomonas citri is a notorious disease affecting a decrease in fruit productivity and quality. Citrus export to USA is also prohibited by the disease. Therefore, development of citrus canker resistant variety is essential and exploitation of markers for molecular breeding is urgent. To develop DNA molecular markers, we performed whole genome resequencing for 8 varieties: 4 citrus canker resistant varieties including C. hybrid ‘Kioymi’ and 4 citrus canker susceptible varieties including C. iyo ‘Miyauchiiyokan’. In total, 642 polymorphic SNPs were detected between resistant and susceptible varieties. Of the 642 SNPs, 50 SNPs were preferably selected based on integrative genomics viewer. To apply the markers in a broad range of citrus variety, we performed genotyping with 6 other varieties very well known as citrus canker resistant and susceptible varieties in addition to previous mentioned 8 varieties. Three of the 50 SNPs were identified as a marker to distinguish citrus canker resistant varieties from susceptible varieties. Secondly, we developed molecular markers to apply for F1 lines crossed by ‘Kiyomi’ and ‘Miyauchiiyokan’. Of the 50 SNPs, we identified 2 SNP markers to distinguish between F1 resistant and susceptible lines. One of them is a resistance gene that plays a role in plant defense mechanism. In this study, we developed 5 molecular marker candidates possible to apply for molecular breeding to develop citrus canker resistant variety. We are working on development of candidate markers related to citrus canker.

Citrus canker caused by Xanthomonas citri pv. citri is one of economically important diseases in the citrus industry. The devastating bacterial disease results in unattractive quality and a significant reduction in fruit production. Citrus growers and industry in Korea has been struggling with the serious disease causing the prohibition of export market. Korea also became the top import market for oranges. The development of markers linked to citrus canker resistance is strongly needed. In this study, we investigated molecular markers between ‘Kiyomi’ (Citrus unshiu ｘ C. sinensis), a resistant cultivar, and Natsudaidai (C. natsudaidai), a susceptible cultivar. To develop markers, we focused on structural variation (copy number variation, CNV, and presence/absence variation, PAV). It has been well documented that CNV and PAV of defense-related genes are associated with resistant cultivars. Using a read depth approach following next-generation sequencing, we performed genome-wide analysis of CNV and PAV in two varieties. As a result, 633 genes showing at least two times difference between the mapping reads from two varieties and 61 genes showing presence of the mapping reads in only either one of them were screened. Visual inspection using the Integrative Genomics Viewer (IGV) was performed and experimental validation is being investigated. Interestingly, one of PAV candidates showed polymorphism in ‘Kiyomi’ and ‘Natsudaidai’ as well as other resistant and susceptible cultivars. Our results suggest a necessity for the detection of structural variation and indicate that the candidates may be useful for molecular breeding for citrus canker resistance and understanding disease resistance mechanism.

TCR subunits are members of membrane-bound receptors which allow the fast and efficient elimination of the specific fish pathogens have regulated function in adaptive immunity. Sequence structure of TCR subunits have been reported for various teleosts, but the information of each TCR subunit functional characterization through expression analysis in fish was unknown. In this study, we examined the gene expression of TCR subunits in the early developmental stages and observed transcript levels in various tissues from healthy adult olive flounder by RT-PCR. The mRNA expression of alpha subunit was already detected in the previous hatching step. But the transcripts of another TCR subunit were not observed during embryo development and increased after hatching and maintained until metamorphosis at the same level. It was found that all TCR subunits mRNAs are commonly expressed in the immune-related organ such as spleen, kidney and gill, also weak expressed in fin and eye. TCR alpha and beta subunit were expressed in brain, whereas gamma and delta were not expressed same tissue. The sequence alignment analysis shows that there are more than 80% sequence homology between TCR subunits. Because it has a high similarity of amino acid sequence to expect similar in function, but expression analysis show that will have may functional diversity due to different time and place of expression.

The innate immune system is the only defense weapon that invertebrates have, and it is the fundamental defense mechanism for fish. The innate immune response is important in newly hatched flounders because it is closely involved in the initial feeding phase, which is why it is essential for survival during the juvenile period. The expression analysis of genes involved in the innate immune response in the olive flounder (Paralichthys olivaceus) in the days after hatching is incomplete. Therefore, we have begun to examine the expression patterns of genes specifically induced during the development of the innate immune system in newly hatched flounders. Microscopic observation showed that pronephron formation corresponded with the expression of perforin-encoding gene. These results suggest that perforin plays a vital role in the innate immunity of the kidney during developmental stages. Perforin expression was strong at the start of the development of the innate immune response, and continued throughout all the development stages. Our findings have important implications with respect to perforin’s biological role and the evolution of the first defense mechanisms in olive flounder. Further studies are required to elucidate the perforin-mediated innate immunity response and to decipher the functional role of perforin in developmental stages.

Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival in such an environment. The key component of innate immune system, C3 is central protein of all activation pathways of the complement system, leading to inflammatory reactions, such as opsonisation, chemotaxis, and cell lysis of pathogens. Although, innate mechanisms is essential for survival in the early stage of development, little is known about defence mechanisms. In this study, the alignment analysis showed that amino acid sequence of C3 from olive flounder liver EST homologous to other known C3 sequences with 73-99% identity. Also, we examined the tissue distribution of olive flounder C3 and analyzed expression pattern from the fertilized egg until 28 days post hatching. As a result, olive flounder C3 mRNA was expressed only in the liver and the mRNA level more increased as developmental proceed during the early stage. These results may suggest that olive flounder C3 plays an important function in the early immune response of olive flounder larvae.