• Journal of Environmental Science International
• /
• v.30 no.5
• /
• pp.379-389
• /
• 2021

Soil microorganism activity in an agricultural field is affected by various factors including climate conditions, soil chemical properties, and crop cultivation. In this study, we elucidate the correlation between microorganism activity and agricultural environment factors using the dehydrogenase activity (DHA) value, which is one of the indicators of soil microbial activity. As a result, the various factors noted above were related to the DHA value. Annual rainfall, soil Mg2+, bacterial and fungal diversities, types of crops, developmental stages, seasons, and cultivation status were highly correlated with the DHA value. Furthermore, next-generation sequencing (NGS) analysis was used to identify that the type of crop affected soil microbial compositions of both bacteria and fungi. Soil used for soybean cultivation showed the highest relative abundance for Verrucomicrobia, Planctomycetes, and Acidobacteria but Actinobacteria and Firmicutes had the lowest relative abundance. In the case of soil used for potato cultivation, Actinobacteria had the highest relative abundance but Proteobacteria had the lowest relative abundance. Armatimonadetes showed the highest relative abundance in soil used for cabbage cultivation. Among the fungal communities, Mortierellomycota had the highest relative abundance for soybean cultivation but the lowest relative abundance for cabbage cultivation; further, Rozellomycota, Chytridiomycota, and Cercozoa had the highest relative abundance for cabbage cultivation. Basidiomycota had the highest relative abundance for potato cultivation but the lowest relative abundance for soybean cultivation.

• Journal of Genetic Medicine
• /
• v.19 no.1
• /
• pp.14-21
• /
• 2022

Complex chromosome rearrangements (CCRs) are structural chromosomal rearrangements involving at least three chromosomes and more than two breakpoints. CCR carriers are generally phenotypically normal but related to higher risk of recurrent miscarriage and having abnormal offspring with congenital anomalies. However, most of CCR carriers are not aware of their condition until genetic analysis of either abortus or affected baby or parental karyotyping is performed. Herein, we present the case that CCR carrier patients can be identified by preimplantation genetic testing of preimplantation embryos. An infertile male patient with severe oligoasthenoteratozoospermia was diagnosed balanced reciprocal translocation, 46,XY,t(3;11) (p26;p14) at first. After attempting the first preimplantation genetic testing for structural rearrangement (PGT-SR) cycle, we found the recurrent segmental gain or loss on 21q21.3-q22.3 of five out of nine embryos. As a result of karyotype re-analysis, the patient's karyotype showed a balanced CCR involving chromosomes 3, 11, and 21 with three breakpoints 3p26, 11p14, and 21q21. The patient underwent two PGT-SR cycles, and a pregnancy was established after the transfer of an euploid embryo in the second cycle. Amniocentesis confirmed that the baby carried normal karyotype without mosaicism. At 37 weeks gestation, a healthy girl weighting 3,050 g was born.

• Parasites, Hosts and Diseases
• /
• v.61 no.2
• /
• pp.183-193
• /
• 2023

Balamuthia mandrillaris amebic encephalitis (BAE) can cause a fatal condition if diagnosis is delayed or effective treatment is lacking. Patients with BAE have been previously reported in 12 provinces of China, with skin lesions being the primary symptom and encephalitis developing after several years. However, a significantly lower number of cases has been reported in Southwest China. Here we report an aggressive BAE case of a 64-year-old woman farmer with a history of skin lesions on her left hand. She was admitted to our hospital due to symptoms of dizziness, headache, cough, vomiting, and gait instability. She was initially diagnosed with syphilitic meningoencephalitis and received a variety of empirical treatment that failed to improve her symptoms. Finally, she was diagnosed with BAE combined with amebic pneumonia using next-generation sequencing (NGS), qRT-PCR, sequence analysis, and imaging studies. She died approximately 3 weeks after the onset. This case highlights that the rapid development of encephalitis can be a prominent clinical manifestation of Balamuthia mandrillaris infection.

• Journal of Life Science
• /
• v.30 no.7
• /
• pp.625-629
• /
• 2020

The Ocean is a rich source of diverse living organisms include viruses. In this study, we examined the microbial communities in the sea adjacent to Jeju Island in two seasons by metatranscriptomics. We collected and extracted total RNA, and, using the next-generation sequencing HiSeq 2000 and de novo transcriptome assembly, we identified 652,984 and 163,759 transcripts from the March and December samples, respectively. The most abundant organisms in March were bacteria, while eukaryotes were dominant in the December sample. The bacterial communities differed between the two samples, suggesting seasonal change. To identify the viruses, we searched the transcripts against a viral reference database using MegaBLAST with the most identified being bacteriophages infecting the marine bacteria. However, we also revealed an abundance of transcripts associated with diverse herpesviruses in the two transcriptomes, indicating the presence or possible threat of infection of fish in the sea around Jeju Island. This data is valuable for the study of marine microbial communities and for identifying possible viral pathogens.

• Korean Journal of Ecology and Environment
• /
• v.56 no.1
• /
• pp.36-44
• /
• 2023

Environmental DNA (eDNA) can exist in both intracellular and extracellular forms in natural ecosystems. When targeting harmful cyanobacteria, extracellular eDNA indicates the presence of traces of cyanobacteria, while intracellular eDNA indicates the potential for cyanobacteria to occur. However, identifying the "actual" potential for harmful cyanobacteria to occur is difficult using the existing sediment eDNA analysis method, which uses silica beads and cannot distinguish between these two forms of eDNA. This study analyzes the applicability of a density gradient centrifugation method (Ludox method) that can selectively analyze intracellular eDNA in sediment to overcome the limitations of conventional sediment eDNA analysis. PCR was used to amplify the extracted eDNA based on the two different methods, and the relative amount of gene amplification was compared using electrophoresis and Image J application. While the conventional bead beating method uses sediment as it is to extract eDNA, it is unknown whether the mic gene amplified from eDNA exists in the cyanobacterial cell or only outside of the cell. However, since the Ludox method concentrates the intracellular eDNA of the sediment through filtration and density gradient, only the mic gene present in the cyanobacteria cells could be amplified. Furthermore, the bead beating method can analyze up to 1 g of sediment at a time, whereas the Ludox method can analyze 5 g to 30 g at a time. This gram of sediments makes it possible to search for even a small amount of mic gene that cannot be searched by conventional bead beating method. In this study, the Ludox method secured sufficient intracellular gene concentration and clearly distinguished intracellular and extracellular eDNA, enabling more accurate and detailed potential analysis. By using the Ludox method for environmental RNA expression and next-generation sequencing (NGS) of harmful cyanobacteria in the sediment, it will be possible to analyze the potential more realistically.

• Journal of Plant Biotechnology
• /
• v.42 no.4
• /
• pp.312-325
• /
• 2015

In this review, we summarized the trends of genomics and transcriptomics research on peach, a model species of Rosaceae. Peach genome maps have been developed from various progeny groups with many next-generation sequencing (NGS) based single nucleotide polymorphism markers. Molecular markers of qualitative traits and quantitative trait loci (QTL) such as fruit characteristics, blooming date, and disease resistance have been analyzed. Among many characteristics, markers related to flesh softening and flesh adhesion are useful for marker assisted selection. Through comparative genomics, peach genome has been compared to the genome of Arabidopsis, Populus, Malus, and Fragaria species. Through transcriptomics and proteomics, fruit growth and development, and flavonoid synthesis, postharvest related transcriptomes and disease resistance related proteins have been reported. Recently, development of NGS based markers, construction of core collection of germplasm, and genotyping of various progenies have been preceded. In the near future, accurate QTL analysis and identification of useful genes are expected to establish a foundation for effective molecular breeding.

• Journal of Plant Biotechnology
• /
• v.44 no.4
• /
• pp.372-378
• /
• 2017

For comparison of the transcription profiles in apple (Malus domestica L.) cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red flesh apple cultivar, 'Redfield' and white flesh apple cultivar, 'Granny Smith' were investigated by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the red flesh apple cultivar and white flesh apple cultivar were selected for nucleotide sequence determination and homology searches. High resolution melting (HRM) technique measures temperature induced strand separation of short PCR amplicons, and is able to detect variation as small as one base difference between red flesh apple cultivars and white flesh apple cultivars. We applied high resolution melting (HRM) analysis to discover single nucleotide polymorphisms (SNP) based on the predicted SNP information derived from the apple EST database. All 103 pairs of SNPs were discriminated, and the HRM profiles of amplicons were established. Putative SNPs were screened from the apple EST contigs by HRM analysis displayed specific difference between 10 red flesh apple cultivars and 11 white flesh apple cultivars. In this study, we report an efficient method to develop SNP markers from an EST database with HRM analysis in apple. These SNP markers could be useful for apple marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in apple cultivars.

• Korean Journal of Plant Resources
• /
• v.32 no.4
• /
• pp.303-311
• /
• 2019

Various colors of fruit skin and flesh are the most popular commercial criteria for peach classification. In order to breed new red-fleshed peach cultivar, many cross seedlings and generations should be maintained. Therefore it is necessary to develop early selection markers to screen seedlings with target traits to increase breeding efficiency. For the comparison of transcription profiles in peach cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red-fleshed peach cultivar, 'Josanghyeoldo' and white-fleshed peach cultivar, 'Mibaekdo' were analyzed by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the two cultivars were selected for nucleotide sequence determination and homology searches. Putative single nucleotide polymorphisms (SNP) were screened from peach EST contigs by high resolution melting (HRM) analysis displayed specific difference between 8 red-fleshed peach cultivars and 24 white-fleshed peach cultivars. All 72 pairs of SNPs were discriminated and the HRM profiles of amplicons were established. In the study reported here, the development of SNP markers for distinguishing between red and white fleshed peach cultivars by HRM analysis offers the opportunity to use DNA markers. This SNP marker could be useful for peach marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in peach cultivars.

• 한국균학회소식:학술대회논문집
• /
• 2015.05a
• /
• pp.41-41
• /
• 2015

BSA-seq technologies, combined Bulked Segregant Analysis (BSA) and Next-Generation Sequencing (NGS), are making it faster and more efficient to establish the association of agronomic traits with molecular markers or candidate genes, which is the requirement for marker-assisted selection in molecular breeding. Clubroot disease, caused by Plasmodiophora brassicae, is a serious threat to Brassica crops. Even we have breed new clubroot resistant varieties of Chinese cabbage (B. rapa ssp. pekinesis), the underlying genetic mechanism is unclear. In this study, an $F_2$ population of 340 plants were inoculated with P. brassicae from Xinye (Pathotype 2 on the differentials of Williams). Resistance phenotype segregation ratio for the populations fit a 3:1 (R:S) segregation model, consistent with a single dominant gene model. Super-BSA, using re-sequencing the parents, extremely R and S DNA pools with each 50 plants, revealed 3 potential candidate regions on the chromosome A03, with the most significant region falling between 24.30 Mb and 24.75 Mb. A linkage map with 31 markers in this region was constructed with several closely linked markers identified. A Major QTL for clubroot resistance, CRq, which was identified with the peak LOD score at 169.3, explaining 89.9% of the phenotypic variation. And we developed a new co-segregated InDel marker BrQ-2. Joint BSA-seq and traditional QTL analysis delimited CRq to an 250 kb genomic region, where four TIR-NBS-LRR genes (Bra019409, Bra019410, Bra019412 and Bra019413) clustered. The CR gene CRq and closely linked markers will be highly useful for breeding new resistant Chinese cabbage cultivars.

• Journal of Plant Biotechnology
• /
• v.47 no.2
• /
• pp.141-149
• /
• 2020

Solanum hougasii, one of the wild Solanum species, has been widely used in potato breeding since it exhibits excellent resistance to diverse important pathogens. S. hougasii can be directly crossed with the cultivated tetraploid potato (S. tuberosum) owing to its EBN (Endosperm Balanced Number) value of 4, which is same as that of S. tuberosum although it is an allohexaploid. In this study, the complete chloroplast genome sequence of S. hougasii was obtained by next-generation sequencing technology, and compared with that of the chloroplast genome of seven other Solanum species to identify S. hougasii-specific PCR markers. The length of the complete chloroplast genome of S. hougasii was 155,549 bp. The structural organization of the chloroplast genome in S. hougasii was found to be similar to that of seven other Solanum species studied. Phylogenetic analysis of S. hougasii with ten other Solanaceae family members revealed that S. hougasii was most closely related to S. stoloniferum, followed by S. berthaultii, and S. tuberosum. Additional comparison of the chloroplast genome sequence with that of five other Solanum species revealed five InDels and 43 SNPs specific to S. hougasii. Based on these SNPs, four PCR-based markers were developed for the differentiation of S. hougasii from other Solanum species. The results obtained in this study will aid in exploring the evolutionary and breeding aspects of Solanum species.