한국작물학회 2017년도 9th Asian Crop Science Association conference
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pp.98-98
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2017
Leaf senescence is the process of aging in plants. Chlorophyll degradation during leaf senescence has the important role translocating nutrients from leaves to storage organs. The functional stay-green with slow leaf yellowing and photosynthesis activity maintenance has been considered one of strategy for increasing crop productivity. Here, we have identified two QTLs on chromosome 9 and 10 for leaf senescence with chlorophyll content of RIL population derived from a cross between Hanareum 2, early leaf senescence Indica-type variety, and Unkwang, delayed leaf senescence Japonica variety. Among these QTLs, we chose qPLS1 QTL on chromosome 9 for further study. qPLS1 was found to explain 14.4% of the total phenotypic variation with 11.2 of LOD score. Through fine-mapping approach, qPLS1 QTL locus was narrowed down to about 25kb in the marker interval between In/del-4-7-9 and In/del-5-9-4. There are 3 genes existed within 25kb of qPLS1 locus: LOC_Os09g36200, LOC_Os09g36210, and LOC_Os09g36220. Among these genes, transcript level of LOC_Os09g36200 was increased during the leaf senescence stage and the expression level of LOC_Os09g36200 in Indica was higher than in Japonica. Finally, we chose LOC_Os09g36200 as candidate gene and renamed it as OsPLS1-In and OsPLS1-Jp from Indica- and Japonica-type rice, respectively. OsPLS1-In and OsPLS1-Jp overexpressing transgenic plants showed both early leaf senescence phenotype. These results indicate that OsPLS1 functions in chlorophyll degradation and the difference of expression level of OsPLS1 cause the difference of leaf senescence between Indica and Japonica in rice.
The mature wheat embryo is arguably one of the best explants for genetic transformation because of its unlimited availability and lack of growth season restriction. However, an efficient regeneration system using mature wheat embryos (Triticum aestivum L.) is still not available. To identify genes related to the tissue culture response (TCR) of wheat, QTLs for callus induction from mature embryos and callus regeneration were mapped using an RIL population derived from the cross of 'Wangshuibai' with 'Nanda2419', which has a good TCR. By whole genome scanning we identified five, four and four chromosome regions conditioning, respectively, percent embryos forming a callus (PEFC), percent calli regenerating plantlets (PCRP), and number of plantlets per regenerating callus (NPRC). The major QTLs QPefc.nau-2A and QPcrp.nau-2A were mapped to the long arm of chromosome 2A, explaining up to 22.8% and 17.6% of the respective phenotypic variance. Moreover, two major QTLs for NPRC were detected on chromosomes 2D and 5D; these together explained 51.6% of the phenotypic variance. We found that chromosomes 2A, 2D, 5A, 5B and 5D were associated via different intervals with at least two of the three TCR indexes used. Based on this study and other reports, the TCRs of different explant types of wheat may be under the control of shared or tightly linked genes, while different genes or gene combinations may govern the stages from callus induction to plantlet regeneration. The importance of group 2 and 5 chromosomes in controlling the TCRs of Triticeae crops and the likely conservation of the corresponding genes in cereals are discussed.
한국생물정보시스템생물학회 2001년도 제2회 생물정보 워크샵 (DNA Chip Bioinformatics)
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pp.61-86
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2001
All cancers are caused by abnormalities in DNA sequence. Throughout life, the DNA in human cells is exposed to mutagens and suffers mistakes in replication, resulting in progressive, subtle changes in the DNA sequence in each cell. Since the development of conventional and molecular cytogenetic methods to the analysis of chromosomal aberrations in cancers, more than 1,800 recurring chromosomal breakpoints have been identified. These breakpoints and regions of nonrandom copy number changes typically point to the location of genes involved in cancer initiation and progression. With the introduction of molecular cytogenetic methodologies based on fluorescence in situ hybridization (FISH), namely, comparative genomic hybridization (CGH) and multicolor FISH (m-FISH) in carcinomas become susceptible to analysis. Conventional CGH has been widely applied for the detection of genomic imbalances in tumor cells, and used normal metaphase chromosomes as targets for the mapping of copy number changes. However, this limits the mapping of such imbalances to the resolution limit of metaphase chromosomes (usually 10 to 20 Mb). Efforts to increase this resolution have led to the "new"concept of genomic DNA chip (1 to 2 Mb), whereby the chromosomal target is replaced with cloned DNA immobilized on such as glass slides. The resulting resolution then depends on the size of the immobilized DNA fragments. We have completed the first draft of its Korean Genome Project. The project proceeded by end sequencing inserts from a library of 96,768 bacterial artificial chromosomes (BACs) containing genomic DNA fragments from Korean ethnicity. The sequenced BAC ends were then compared to the Human Genome Project′s publicly available sequence database and aligned according to known cancer gene sequences. These BAC clones were biotinylated by nick translation, hybridized to cytogenetic preparations of metaphase cells, and detected with fluorescein-conjugated avidin. Only locations of unique or low-copy Portions of the clone are identified, because high-copy interspersed repetitive sequences in the probe were suppressed by the addition of unlabelled Cotl DNA. Banding patterns were produced using DAPI. By this means, every BAC fragment has been matched to its appropriate chromosomal location. We have placed 86 (156 BAC clones) cytogenetically defined landmarks to help with the characterization of known cancer genes. Microarray techniques would be applied in CGH by replacement of metaphase chromosome to arrayed BAC confirming in oncogene and tumor suppressor gene: and an array BAC clones from the collection is used to perform a genome-wide scan for segmental aneuploidy by array-CGH. Therefore, the genomic DNA chip (arrayed BAC) will be undoubtedly provide accurate diagnosis of deletions, duplication, insertions and rearrangements of genomic material related to various human phenotypes, including neoplasias. And our tumor markers based on genetic abnormalities of cancer would be identified and contribute to the screening of the stage of cancers and/or hereditary diseases
As a first step towards identifying genes involving in the signal transduction pathways mediating rice blast resistance, we isolated 3 mutants lines that showed enhanced susceptibility to rice blast KJ105 (91-033) from a T-DNA insertion library of the japonica rice cultivar, Hwayeong. Since none of the susceptible phenotypes co-segregated with the T-DNA insertion we adapted a map-based cloning strategy to isolate the gene(s) responsible for the enhanced susceptibility of the Hwayeong mutants. A genetic mapping population was produced by crossing the resistant wild type Hwayeong with the susceptible cultivar, Nagdong. Chi-square analysis of the $F_2$ segregating population indicated that resistance in Hwayeong was controlled by a single major gene that we tentatively named Pi-hy. Randomly selected susceptible plants in the $F_2$ population were used to build an initial map of Pi-hy. The SSLP marker RM2265 on chromosome 2 was closely linked to resistance. High resolution mapping using 105 $F_2$ plants revealed that the resistance gene was tightly linked, or identical, to Pib, a resistance gene with a nucleotide binding sequence and leucine-rich repeats (NB-LRR) previously isolated. Sequence analysis of the Pib locus amplified from three susceptible mutants revealed lesions within this gene, demonstrating that the Pi-hy gene is Pib. The Pib mutations in 1D-22-10-13, 1D-54-16-8, and 1C-143-16-1 were, respectively, a missense mutation in the conserved NB domain 3, a nonsense mutation in the 5th LRR, and a nonsense mutation in the C terminus following the LRRs that causes a small deletion of the C terminus. These findings provide evidence that NB domain 3 and the C terminus are required for full activity of the plant R gene. They also suggest that alterations of the resistance gene can cause major differences in pathogen specificity by affecting interactions with an avirulence factor.
Kim, Tae-Hun;Choi, Bong-Hwan;Yoon, Du-Hak;Park, Eung-Woo;Jeon, Jin-Tae;Han, Jae-Young;Oh, Sung-Jong;Cheong, Il-Cheong
Asian-Australasian Journal of Animal Sciences
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제17권9호
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pp.1210-1213
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2004
Quantitative trait loci (QTL) mapping can be applied to detect chromosomal locations that control economic traits in farm animals. Teat number has been considered as one of the most important factors to evaluate mothering ability of sow. Especially, teat number is more important when the number is less than the litter size. This study was conducted to identify QTL affecting teat number in the Korean native pig${\times}$Landrace resource family. A total of 240 animals was genotyped for 132 polymorphic microsatellites covering the 18 pig autosomes. Mean and standard deviation of teat number in $F_2$animals is 13.46${\pm}$1.40. QTL was analyzed using F2 QTL Analysis Servlet of QTL express. A QTL for teat number on SSC9 was significant at the 1% chromosome-wide level and three suggestive QTL were detected on SSC3, 7 and 14. All QTL detected in this study had additive effect and Landrace alleles were associated with higher teat number in comparison with Korean native pig for three of four QTL.
This study was carried out to map Bphl and bph2 gene in Mudgo and Sangju13 (Oryza sativa L.) respectively conferring resistance to brown plan-thopper (BPH) and to establish the marker-assisted selection (MAS) system. Bulked seedling (grown for 20 days) test was conducted with the 73 F4 lines derived from a cross between Nagdongbyeo and Mudgo for Bphl and with 53 BC3F5 lines derived from the Milyang95/Sangju13 cross for bph2. Bph1 was mapped between RG413 and RG901 on chromo-some 12 at a distance of 7.5 cM from RG413 and 8.4 cM from RG90l. A recessive gene bph2 was located near RZ76 on chromosome 12 at a distance of 14.4 cM. Bphl and bph2 were linked to each other with a distance of about 30 cM. An RFLP marker, RG413 linked to Bphl, was converted to an STS marker to facilitate the marker-assisted selection. BPH resistant genotypes could be selected with 92% accuracy in a population derived from a line of NM47-B-B.
International Journal of Industrial Entomology and Biomaterials
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제18권2호
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pp.113-120
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2009
Embryonic lethal abnormal visual (elav) is a lethal gene in Drosophila inducing the abnormal development and function of nervous system. We cloned a Bm-elav gene by bioinformatics and biological experiment, based on sequence of ELAV protein and dbEST of Bombyx mori. The full-length of Bm-elav cDNA is 1498 bp, contains a 906 bp open read frame (ORF) encoding a precursor of 301 amino acid residues with a calculated molecular weight of 34 kDa and pI of 8.99. Bm-ELAV protein precursor contains three RNA recognition motifs (RRM) in $24{\sim}91$, $110{\sim}177$ and $222{\sim}295$ bit amino acid residues respectively, and belongs to RNA-binding protein family. Bm-ELAV shared varying positives, ranging from 56% to 60% (Identities from 41% to 45%), with RRM from other species of Xenopus tropicalis, Apis mellifera, Tribolium castaneum, Branchiostoma belcheri and Drosophila. Gene localization indicated that Bm-elav is a single-copy gene, gene mapping within 12-chromosome from 7916.68 knt to 7918.16 knt region of nscaf2993. Spatiotemporal expressions pattern analysis revealed that Bm-elav expressed higher in most tested tissues and developmental stages in whole generation, such as silk gland, fat body, midgut, hemopoietic organ and ovary, but almost no expression in terminated diapause eggs. This suggested that the expression of Bm-elav in early developmental embryonic stages might induce abnormal development like in Drosophila. Cloning of the Bm-elav gene enables us to test its potential role in controlling pests by transferring the gene into field lepidopteran insects in the future.
Wang, Heng;Yang, Shulin;Tang, Zhonglin;Mu, Yulian;Cui, Wentao;Li, Kui
Asian-Australasian Journal of Animal Sciences
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제20권9호
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pp.1349-1353
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2007
Calcineurin is a calmodulin dependent protein that functions as a regulator of muscle cell growth and function. Agents capable of interacting with calcineurin could have important applications in muscle disease treatment as well as in the improvement of livestock production. Calsarcins comprise a family of muscle-specific calcineurin binding proteins which play an important role in modulating the function of calcineurin in muscle cells. Recently, we described the first two members of the calsarcin family (calsarcin-1 and calsarcin-2) in the pig. Here, we characterized the third member of the calsarcin family, calsarcin-3, which is also expressed specifically in skeletal muscle. However, unlike calsarcin-1 and calsarcin-2, the calsarcin-3 mRNA expression in skeletal muscle kept rising throughout the prenatal and postnatal development periods. In addition, radiation hybrid mapping indicated that porcine calsarcin-3 mapped to the distal end of the q arm of pig chromosome 2 (SSC2). A C/T single nucleotide polymorphism site in exon 5 was genotyped using the denaturing high performance liquid chromatography (DHPLC) method and the allele frequencies at this locus were significantly different among breeds.
Kim, Sun Ha;Shim, Kyu-Chan;Lee, Hyun-Sook;Le, Anh Quynh;Ahn, Sang-Nag
한국작물학회:학술대회논문집
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한국작물학회 2017년도 9th Asian Crop Science Association conference
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pp.116-116
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2017
Low-temperature is one of the environmental stress factors that affect plant growth and development and consequently limit crop productivity. The control of seed germination under low-temperature is organized by many genes which are called quantitative trait loci (QTLs). High germination rate for low-temperature is an important factor of growing rice. Previously, we identified a major QTL controlling low-temperature germinability in rice using 96 introgression lines (ILs) derived from a cross between Oryza rufipogon (Rufi) and the Korean japonica cultivar, 'Hwaseongbyeo (HS)'. A $BC_3F_7$ line (TR5) showed better low-temperature germinability than its recurrent parent. TR5 was crossed with HS to develop a segregating F2:3 populations for the target QTL. Six SSR markers polymorphic between HS and Rufi were used to screen and fine map the qLTG1. The qLTG1 on chromosome 1, which accounted for 55.5% of the total phenotypic variation, confirmed that Rufi allele enhanced the low-temperature germinability. Intervals between markers CRM16 and CRM15, four candidate genes were identified. The identified candidate genes, which are encoded by a protein of unknown function, showed their direct involvement on seed germination at low-temperature. To identify genes targeted by qLTG1, we investigated the expression profiles of these candidate genes and germination behavior of qLTG1 under different stress conditions and compared to HS, Rufi, and TR5 at $13{\pm}2^{\circ}C$ for 3 days after incubation. Furthermore, transgenic rice plants will also be developed to conduct a detailed investigation on low-temperature germinability. Hence, the QTL for low-temperature germinability would be useful in rice breeding programs especially in the development of lines possessing low-temperature germinability.
A simulation study was conducted to evaluate the effect of reciprocal cross on the detection and characterization of parent-of-origin (POE) QTL in $F_2$ QTL populations. Data were simulated under two different mating designs. In the one-way cross design, six $F_0$ grand sires of one breed and 30 $F_0$ grand dams of another breed generated 10 $F_1$ offspring per dam. Sixteen $F_1$ sires and 64 $F_1$ dams were randomly chosen to produce a total of 640 $F_2$ offspring. In the reciprocal design, three $F_0$ grand sires of A breed and 15 $F_0$ grand dams of B breed were mated to generate 10 $F_1$ offspring per dam. Eight $F_1$ sires and 32 $F_1$ dams were randomly chosen to produce 10 $F_2$ offspring per $F_1$ dam, totaling 320 $F_2$ offspring. Another mating set comprised three $F_0$ grand sires of B breed and 15 $F_0$ grand dams of A breed to produce the same number of $F_1$ and $F_2$ offspring. A chromosome of 100 cM was simulated with large, medium or small QTL with fixed or different allele frequencies in parental breeds. A series of tests between Mendelian and POE models were applied to characterize QTL as Mendelian, paternal, maternal or partial expression QTL. The overall detection powers were similar between the two mating designs. However, the proportions of paternally expressed QTL that were declared as paternal QTL type were greater in the reciprocal cross design than in the one-way cross, and vice versa for Mendelian QTL. When QTL alleles were segregating in parental breeds, a significant proportion of Mendelian QTL were spuriously declared POE QTL, suggesting that care must be taken to characterize imprinting QTL in a QTL mapping population with a small number of $F_1$ parents.
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