To elucidate the mechanism of pyrethroid resistance in Helicoverpa armigera, the study explored three possibilities based on deltamethrin as a model pyrethroid; 1) the existence of mutations in the target site of deltamethrin, 2) the existence of variation at the genomic level between insecticide-susceptible and resistant strains, 3) differences in gene expression patterns between the strains. Based on these hypotheses, three levels of resistant strains and a susceptible strain as well as nine Korean field populations were used. As results, 1) any point mutations were not detected in sodium channel gene. 2) based on newly set Korean reference genome (GCA_026262555.1), approximately 3,369,837 and 1,032,689 variants (SNPs and Indels) were revealed from genome and ORFs, respectively. However, any specific variants were not found to be highly correlated with the level of insecticide resistance. 3) based on DEG analysis, some of detoxification enzyme genes were differently expressed particularly cytochrome P450 genes. Therefore, H. armigera possibly acquires deltamethrin resistance through a combination of actions, including over-expression of various detoxification enzymes such as CYP3 subfamilies and cuticular proteins.
본 연구는 Yorkshire종, Landrace종 및 Duroc종에 대한 유전체자료를 이용하여 수퇘지의 웅취를 유발하는 세 가지 호르몬인 androstenone, indole 및 skatole 호르몬에 대한 유의적인 유전자영역, SNP 마커 및 후보유전자를 발굴하여 최종적으로 저웅취 종돈을 육종하는데 그 목적이 있다. Genomoe-Wide Association Study를 수행하기 위한 참조집단으로 수집한 유전체 정보는 Yorkshire, Landrace 및 Duroc종에서 각각 3,858 두, 472두 및 1,029두로 총 5,359두에 대한 유전체자료를 분석에 이용하였다. 추정되는 육종가의 정확도를 평가하기 위하여 REML방법을 ASREML 4.1 소프트웨어를 이용하여 분석하였고 세 가지 호르몬에 대하여 다형질 개체모형을 적용하였으며 추정된 육종가로부터 산출한 deregreessed DEBVincPA를 반응변수로 이용하여 연구를 수행하였다. 세 가지 호르몬에 대하여 BayesB와 C의 방법론을 통하여 분석한 결과 BayesB에서 세 가지 호르몬과 연관될 것으로 예상되는 SNP marker 9개, 즉 androstenone에서 3개, indole에서 1개 및 skatole에서 5개가 발굴되었다. BayesC에 서는 이보다 적은 SNP marker 3개가 발굴되었다. 수퇘지의 웅취 호르몬에 영향을 미칠 것으로 예상되는 후보유전자는 총 6개로 각각 LMAN2L, ABLI, NRG3, CDH12, TRAPPC9, MAN1A2로 나타났다
본 연구는 제주지역에서 사육되고 있는 제주 재래돼지기반의 제주흑돈(제주재래돼지 합성돈)에 대하여 산육형질인 등지방 두께, 일당증체량, 90 kg 도달일령에 대한 후보유전자 탐색을 위해 전장유전체 분석을 실시하였고, 유전체 육종가의 정확도 비교를 위해 베이지안 방법(BayesB, BayesC), π(0.00, 0.50, 0.90, 0.99)값과 개체 자신과 후대의 정보만 포함하고 조상의 정보에 대해 정보를 보정한 DEBVexcPA (Deregressed EBV exlude parents average)와 부모 평균을 포함한 DEBVincPA (Deregressed EBV include parents average)의 반응변수를 이용하였다. 산육형질에 대한 전장유전체 분석을 실시한 결과 등지방두께는 KIAA1549 유전자(SSC18 at 11Mb region), 일당증체량과 90 kg 도달일령에서 POLD3(SSC9 at 9Mb region), STX6(SSC9 at 134Mb region) 유전자가 공통적으로 탐색되었다. 유전체의 정확도 범위는 일당증체량 BayesB의 경우 DEBVexcPA와 DEBVincPA에서 각각 0.285~0.305와 0.497~0.510과 BayesC에서 0.287~0.296와 0.499~0.511으로 추정되었다. 90 kg 도달일령은 BayesB에서 DEBVexcPA와 DEBVincPA는 각각 0.284~0.305와 0.497~0.510, BayesC에서는 0.284~0.296와 0.498-0.511로 추정되었다. 특히, 등지방두께의 유전체 정확도가 BayesB방법론의 DEBVincPA에서 0.625로 가장 높게 추정되었다. 제주흑돈의 산육형질에 대한 유전적인 개량 속도를 가속화하기 위해 유전체 선발법을 적용하는데 있어 부모 평균을 포함한 DEBVincPA (Deregressed EBV include parents average)의 반응변수를 이용하는 것이 바람직하다고 판단된다.
Some behaviors of pigs that are not expressed in the wild state or are observed in a small minority of individuals after groups of pigs are mixed have been reported to indicate poor welfare. A GWAS analysis was performed by measuring the frequency and duration of the four ethological traits and using the mlma command provided by the genome-wide complex trait analysis (GCTA). The positional candidate genes on significantly identified single nucleotide polymorphism (SNP) markers were identified by using the dbSNP provided by the National Center for Biotechnology Information (NCBI). When the GWAS analysis was applied the 43,565 (of the purebred Landrace population) and 41,700 (of the purebred Yorkshire population) SNP markers, 1, 2, and 1 significant SNP markers were identified for the traits of feeding frequency (LOC110262254), locomotion time (LOC110260361), and locomotion frequency (LOC110260361) of the purebred Landrace population, respectively. Meanwhile, 1 and 7 significant SNP markers were identified for the traits of drinking time (LOC 110260090) and feeding frequency (MAP3K19; LOC110257013; ACMSD; TMEM163; RAB3GAP1) of the purebred Yorkshire population, respectively. The results of this study may suggest that the GWAS analysis of the ethological traits of purebred Landrace and Yorkshire populations could be used to perform a GWAS analysis on non-economic traits, and the results can thus be provided as basic data for GWAS analyses of other non-economic traits in the future.
According to Livestock Inspection Standards, the piglets enter the feedlot at approximately 30 kg, and the inspection starts after the preliminary feeding period. The reason for applying the preliminary feeding period is to select inspection piglets with no diseases after the complete growth of the internal organs until 10 weeks of age. Furthermore, the age of 10 week is the time when the muscle fibers grow to their maximum size and the piglets are prepared for fat deposition at the later fattening period. In the study, a genome-wide association study (GWAS) was performed through the mlma command of the genome-wide complex trait analysis (GCTA) program with 703 purebred Landrace population, and the candidate genes associated with the weight of 10 week were searched. The GWAS identified 3 single nucleotide polymorphism (SNP) markers, which have a significant genome-wide suggestive level, on chromosome 6 (DIAS0002615; p-value=1.62×10-6, MARC0083933; p-value=4.94×10-6, ASGA0028717; p-value=5.40×10-6). The 2 genes (Ubiquitin protein ligase E3 component n-recognin 4; UBR4, WD and tetratricopeptide repeats 1; WDTC1) in which these 3 SNP markers are located are positional candidate genes of the weight of 10 week of the purebred Landrace population. 2 candidate genes have been reported to be associated with fattening. Therefore, the positional candidate genes in this study, UBR4 and WDTC1, are expected to be usable as genes for traits associated with the weight of 10 week weight and fattening through additional experimental research with other population.
The cotton bollworm, Helicoverpa armigera is one of the most polyphagous and cosmopolitan pest species in world wide including Korea. It also showed high level of resistance against various types of insecticide. To set the integrated resistant management program, first, we tried to identify the insecticide resistant mechanism via transcriptome analysis of coding and non-coding RNAs using 4 strains (Australian susceptible and resistant strains, Korean and Brazilian resistant strains, additional a sub-lethal dose insecticide treatment in Korean resistant strain). From the illumina based RNAseq data sets with genome information, some resistant involved detoxification genes and long non-coding RNAs were discovered. Second, following molecular markers were developed from resistant strain specific amino acid substitution from those candidate genes. Third, functionally identified genes’ markers were applied in field populations using some molecular diagnostic tools such as LAMP (loop mediated isothermal amplification). Now additional functional analysis is going on for these un-characterized candidate genes and long non-coding RNAs.
돼지의 생시체중은 생존율과 폐사율에 밀접한 관련이 있어 양돈산업에서 자돈 관리와 직결된 중요한 경제형질이다. 본 연구는 Genome-Wide Association Study(GWAS) 분석을 통해 순종 랜드레이스의 생 시체중과 관련된 위치상 후보유전자 탐색을 실시하였다. 생시체중의 유의적 관련 분석 결과, genomewide suggestive level에서 유의성 있는 single nucleotide polymorphism(SNP) marker는 3번 염색체 (ASGA0098921, P=2.41×10-5)와 4번 염색체(H3GA0013451, P=2.47×10-5)에서 각각 1개씩 동정되었다. 이들 SNP marker가 위치한 3개의 유전자(LOC110260055, LOC100156472, LOC100157689)들은 순종 랜드레이스 생시체중의 위치상 후보 유전자이며, 이들 유전자 정보를 이용해서 순종 랜드레이스 생시 체중을 선발할 수 있는 기초 연구 자료가 될 것으로 사료된다.
In the swine industry, growth related traits are important economic traits directly linked to profitability. Representative growth traits include daily gain, back fat thickness, and carcass weight. This study was conducted to search for positional candidate genes associated with the carcass weight through a genome-wide association study(GWAS) using suggestive levels of statistical thresholds in pigs. As a result of the genome-wide analysis of the associations with carcass weight, the single nucleotide polymorphism(SNP) markers with suggestive significance were identified in 1 SNP marker on chromosome 2(ALGA0015365) and 1 SNP marker on chromosome 4(ALGA0023678). We could select positional 2 candidate genes, located close to the SNP markers with suggestive significance levels. The SNP markers in adjacent to the 2 genes(LOC100519538, LOC100737583) may provide basic data regarding the marker-assisted selection for the carcass weight trait in pigs.
NGS data was yielded by using Illumina Hiseq platform. The short reads were filtered by quality score and read length were mapped against the reference genome (KACC42780). Genome-wide reanalyzed data of Flammulina strains were compared against the reference genome to establish a genome-wide single nucleotide polymorphism (SNP). The rate of mapping differences between the strains reflected in the strain variation in its result. The genome-wide SNPs distribution divided into types of homozygous SNP and heterozygous SNP moreover all of the strains demonstrated a wide variation in all of the regions. In the further study of topological relationship between the collected strains, phylogenetic tree was separated into 3 major groups. Group I contained F. velutipes var. related strains of ASI 4062, 4148, 4195. Group Ⅱ contained strains that were different species of ASI 4188 F. elastica, ASI 4190 F. fennae, and ASI 4194 F. rossica. The other 19 strains F. velutipes were classified as a single group. Polymorphic SNPs of F. velutipes strains representing the phylogenetic segregation of whiteand brown-fruiting body forming groups were compared. As previously reported, white gene expression is recessive to brown in fruiting body color gene expression. The white strains produced 131,874 SNPs to be aa type and homozygous from of SNP. 407,947 SNPs were detected as AA, Aa type from the brown-fruiting body of SNP. We constructed a SNP matrix with 8 white strains and 12 brown strains. To develop the molecular marker related in to fruiting body color and geographical origin, we isolated 240 SNPs from the white-and brown-fruiting body forming. To determine the chromosome relationship on polymorphic SNP between Korea and Japan strains producing white-fruiting body, we analyzed that the Korea white strains detected 185,695 SNPs and the Japan white strains produced 263,811 SNPs. Using the constructed SNP matrix with 3 Korea white strains and 3 Japan white strains, the experiment generated 475 SNPs of phylogenetic SNPs fromKorea and Japan white-fruiting body. As a result, we regarded as they are potentially related to the white color. White and brown color and origin specific SNPs could be used as an identification marker for selection of F. veluipes strains in the breeding program.
Data on primal cuts were collected from 1,829 steers of Hanwoo progeny testing programs, between 2010 and 2015 for the ssGWAS. SNP data were analyzed by using Illumina Bovine 50K Beadchip. The SNP data that matches with phenotype data was 674 animals. As a first step, the genomic estimated breeding value(GEBV) of the loin and rib cuts were estimated, which was used in the estimation of SNP marker effects and their variances related to the traits. Then, the estimated variance explained by each marker was expressed as a proportion to the total genetic variance. Finally, the SNP loci and their significance to any possible QTL were examined. Among the 20 best SNP loci explaining a larger proportion of SNP variance to the total genetic variance for tender loin yield, the region between 12,812,193 ~ 12,922,313bp on BTA 10 harbored a cluster of SNPs that explained about 7.32 to 7.34% of the total genetic variance. For strip loin yield, a peak for higher effects for multiple SNPs was found in BTA24, between 38,158,543 and 38,347,278bp distances, which explained about 8.36 to 8.56% of the observed variance for this trait. For loin yield had relatively smaller effects in terms of the total genetic variance. Therefore, loin yield might be affected by a few loci with moderate effects and many other loci with smaller effects across the genome.
Copy number variation (CNV) is one of structural variation types that shows various numbers of copies in segments of the DNA. This study aimed to identify the association between copy number variation regions (CNVRs) and carcass traits in Hanwoo. We analyzed a total of 571 Hanwoo steers with the four carcass traits (marbling score (MS), backfat thickness (BF), carcass weight (CW), loineye muscle area (LMA)). PennCNV program was used to identify the CNVs and CNVRuler program was used to analyze the association between CNVRs and carcass traits. A total of 1,659 CNVRs were identified in the whole genome of Hanwoo. These 1,659 CNVRs divided into 415 Gain, 1082 Loss and 162 Gain/Loss events. A genome wide association analysis between the CNVRs and the carcass traits was performed using CNVRuler program. The number of significant CNVR at a threshold of p<1×10-4 was 2, 7, 2 and 1 loci for MS, BF, CW and LMA, respectively. We performed gene ontology (GO) analysis for the genes in the significant CNVRs using DAVID. ABCA2 and EDF1 were related to regulation of lipid metabolic process. C8G, TRAF2 and STAB2 were related to immune. CHST11 was related to developmental growth. Our results may provide an important resource for molecular breeding research in Hanwoo.
The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges. fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylationsites(Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1(Asn-454), fvLac-2(Asn- 437andAsn-455), fvLac-3(Asn-111andAsn-237), and fvLac4 (Asn-402andAsn-457). In addition, the first 19–20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction(RT-PCR) analyses clearly reveal that CuSO4 affects the induction and the transcription level of these laccase genes.
The 35.6-mb genome contained 12,218 predicted protein-encoding genes and 287 tRNA genes assembled into 11 scaffolds corresponding with the 11 chromosomes of strain KACC 42780. Based on presence of copper binding region and signal peptide, 8 laccase genes (Fv Lac-1, Fv Lac-2, Fv Lac-3, Fv Lac-4, Fv Lac-5, Fv Lac-6, Fv Lac-7, Fv Lac-8) were selected in F.velutipes genome. The laccase genes of F.velutipes ranged from 1,300 to 1,700 base pair in their size. In addition, the molecular mass and PI values of laccase genes ranged from 50 to 62 kDa and from 3 to 11, respectively.
In laccase activity assays, the highest laccase activity (3unit/ml)was shown in medium with 0.5mM of CuSO4 on 9th day. However, laccase activity was severely decreased in medium with 1mM and 2mM of CuSO4 was drastically decreased from day 3 through day 9. Interestingly, laccase activity in the medium without CuSO4 showed higher level than in the medium with 0.25, 1 and 2mM on both 6th and 9th day.
RT_PCR showed the highest transcription level with 0.25mM and 0.5mM of CuSO4 supplementation on day 3 and on day 9, respectively. In addition, the transcription level of Lac-1 increased depending on concentration of CuSO4 supplementation.
혈청 내 존재하는 효소 중 Glutamic pyruvic transaminase(GPT)는 근육이나 간세포의 손상에 대한 임상 화학적 지표로 사용 된다. 본 연구는 Landrace와 한국재래돼지의 F2 교잡 축군(N=1,105)에 대해 Porcine SNP 60K beadchip을 사용하여 유전자형 분석을 실시하고, GPT 형질과의 관련성을 검증하기 위해 Genome-Wide Association Study(GWAS)를 수행하였다. F2 교잡축군의 가계구조를 보정한 GWAS를 수행하기 위하여 혼합모형과 회귀분석을 조합한 GRAMMAR방법을 관련성 분석에 사용 하였다. 유의성 있는 SNP 표지들은 Sus scrofa chromosome(SSC) 6, 7, 그리고 13에서 동정되었다. 이들유의성 있는 SNP marker들에 가장 근접한 염색체상 위치의 유전자를 그 유전자의 기능을 고려하여 SSC7에서 2개의 위치후보유전자(BCL11B, AHNAK2)를 선정하였다. Pyrosequencing법을 통하여 이 들유전자 내에 존재하는 4개 SNP 표지들의 유전자형을 분석하여 GPT 형질간의 관련성 분석에 이용하였다. 관련성 분석결과, BCL11B g.267 T>C에서 nominal P=7.23×10-8 과 AHNAK2 g.1439 C>T에서 nominal P=5.64×10-6, g.1736 C>A에서 nominal P=3.51×10-6의 결과를 얻었다. 이 들 중 가장 유의한 결과를 얻은 BCL11B 유전자의 g.267 T>C SNP 표지는 추가 연구를 통하여 혈청 GPT 변이에 영향을 미치는 위치상 후보 유전자 표지로 사용 되어 질 수 있을 것이라 사료되어진다.