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BSA-Seq Technologies Identify a Major QTL for Clubroot Resistance in Chinese Cabbage (Brassica rapa ssp. pekinesis)

  • Yuan, Yu-Xiang;Wei, Xiao-Chun;Zhang, Qiang;Zhao, Yan-Yan;Jiang, Wu-Sheng;Yao, Qiu-Ju;Wang, Zhi-Yong;Zhang, Ying;Tan, Yafei;Li, Yang;Xu, Qian;Zhang, Xiao-Wei
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.41-41
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    • 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.

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Effects of Light Intensity on Photosynthetic Activity of Shade Tolerant and Intolerant Tree Species (광도(光度)가 내음성(耐陰性)이 서로 다른 3수종(樹種)의 광합성(光合成) 생리(生理)에 미치는 영향(影響))

  • Kim, Pan-Gi;Yi, Yong-Sub;Chung, Dong-Jun;Woo, Su-Young;Sung, Joo Han;Lee, Eun Ju
    • Journal of Korean Society of Forest Science
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    • v.90 no.4
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    • pp.476-487
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    • 2001
  • In this study, three tree species (Populus euramericana, Kalopanax pictus and Quercus serrata) exhibiting different levels of shade tolerance were employed to investigate photosynthetic responses to the lower light condition on forest floors. Chlorophyll contents, spectral properties and photosynthetic characteristics were examined by using the tree species grown under high light intensity (PPFD $920{\mu}mol\;m^{-2}s^{-1}$) or low light intensity (PPFD $80{\mu}mol\;m^{-2}s^{-1}$). Plants grown under the low light intensity tended to have reduced leaf area, chlorophyll content per unit leaf area, light absorption and respiration in the shade intolerant tree species of P. euramericana. However, the shade tolerant species K. pictus and Q. serrata showed increased leaf area, chlorophyll content and net apparent quantum yield. Therefore, light absorption and light utilization efficiency were improved under the low light intensity. Also shade tolerant species maintained activities of photosystems and $CO_2$ fixation systems with little changes in low light intensity condition.

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Comparison of Viability, ATP and In vitro Fertilization of Boar Sperm Stored at 4℃ in the Three Different Diluents

  • Yi, Y.J.;Li, Z.H.;Kim, E.S.;Song, E.S.;Cong, P.Q.;Zhang, Y.H.;Lee, S.H.;Lee, J.W.;Park, Chang-Sik
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.8
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    • pp.1127-1133
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    • 2008
  • This study was designed to develop a method of liquid storage of boar sperm at $4^{\circ}C$ by using the modified Beltsville F5 (BF5) diluent with bovine serum albumin (BSA) and N-acetyl-D-glucosamine. Boar sperm were stored in lactose-egg yolk and N-acetyl-D-glucosamine (LEN), BF5 and Golden-Pig liquid 4 (GPL4) diluents at $4^{\circ}C$ for 5 days and were examined for sperm viability, adenosine triphosphate (ATP) and in vitro fertilization (IVF). The percentage of sperm viability in GPL4 diluent was higher than in LEN and BF5 diluent from 1 to 5 days of storage at $4^{\circ}C$. The percentage of sperm viability steadily declined from 1 to 5 days of storage in the three different diluents. Sperm ATP in GPL4 diluent was higher than in LEN and BF5 diluents from 1 to 5 days of storage. Sperm ATP rapidly declined after 5 days of storage in the three different diluents. Porcine oocytes matured in vitro were inseminated with different sperm concentrations of liquid semen stored for 3 days in GPL4 diluent. The percentage of monospermic oocytes did not show any differences from 2.5 to $20{\times}10^5$ sperm/ml. However, the percentage of polyspermic oocytes in the sperm concentration of $2.5{\times}10^5$ sperm/ml was lower than in concentrations of 5, 10 and $20{\times}10^5$5 sperm/ml. The percentage of blastocysts from the cleaved oocytes at $2.5{\times}10^5/ml$ sperm concentration was significantly lower than at 5, 10 and $20{\times}10^5sperm/ml$ concentrations. In conclusion, GPL4 diluent can be stored at $4^{\circ}C$ for 5 days and showed higher sperm viability and sperm ATP concentration compared with LEN and BF5 diluents. Also, we found that GPL4 diluent can be used for IVF of porcine oocytes.

Sequencing, Genomic Structure, Chromosomal Mapping and Association Study of the Porcine ADAMTS1 Gene with Litter Size

  • Yue, K.;Peng, J.;Zheng, R.;Li, J.L.;Chen, J.F.;Li, F.E.;Dai, L.H.;Ding, SH.H.;Guo, W.H.;Xu, N.Y.;Xiong, Y.ZH.;Jiang, S.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.7
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    • pp.917-922
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    • 2008
  • A disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif (ADAMTS1) plays a critical role in follicular rupture and represents a major advance in the proteolytic events that control ovulation. In this study, a 9,026-bp DNA sequence containing the full coding region, all 8 introns and part of the 5'and 3' untranslated region of the porcine ADAMTS1 gene was obtained. Analysis of the ADAMTS1 gene using the porcine radiation hybrid panel indicated that pig ADAMTS1 is closely linkage with microsatellite marker S0215, located on SSC13q49. The open reading frame of its cDNA covered 2,844 bp and encoded 947 amino acids. The coding region of porcine ADAMTS1 as determined by sequence alignments shared 85% and 81% identity with human and mouse cDNAs, respectively. The deduced protein contained 947 amino acids showing 85% sequence similarity both to the human and mouse proteins, respectively. Comparative sequencing of three pig breeds revealed one single nucleotide polymorphism (SNP) within exon 7 of which a G-C substitution at position 6006 changes a codon for arginine into a codon for proline. The substitution was situated within a PvuII recognition site and developed as a PCR-RFLP marker for further use in population variation investigations and association analysis with litter size. Allele frequencies of this SNP were investigated in seven pig breeds/lines. An association analysis in a new Qingping female line suggested that different ADAMTS1 genotypes have significant differences in litter size (p<0.01).

Characterizing Milk Production Related Genes in Holstein Using RNA-seq

  • Seo, Minseok;Lee, Hyun-Jeong;Kim, Kwondo;Caetano-Anolles, Kelsey;Jeong, Jin Young;Park, Sungkwon;Oh, Young Kyun;Cho, Seoae;Kim, Heebal
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.343-351
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    • 2016
  • Although the chemical, physical, and nutritional properties of bovine milk have been extensively studied, only a few studies have attempted to characterize milk-synthesizing genes using RNA-seq data. RNA-seq data was collected from 21 Holstein samples, along with group information about milk production ability; milk yield; and protein, fat, and solid contents. Meta-analysis was employed in order to generally characterize genes related to milk production. In addition, we attempted to investigate the relationship between milk related traits, parity, and lactation period. We observed that milk fat is highly correlated with lactation period; this result indicates that this effect should be considered in the model in order to accurately detect milk production related genes. By employing our developed model, 271 genes were significantly (false discovery rate [FDR] adjusted p-value<0.1) detected as milk production related differentially expressed genes. Of these genes, five (albumin, nitric oxide synthase 3, RNA-binding region (RNP1, RRM) containing 3, secreted and transmembrane 1, and serine palmitoyltransferase, small subunit B) were technically validated using quantitative real-time polymerase chain reaction (qRT-PCR) in order to check the accuracy of RNA-seq analysis. Finally, 83 gene ontology biological processes including several blood vessel and mammary gland development related terms, were significantly detected using DAVID gene-set enrichment analysis. From these results, we observed that detected milk production related genes are highly enriched in the circulation system process and mammary gland related biological functions. In addition, we observed that detected genes including caveolin 1, mammary serum amyloid A3.2, lingual antimicrobial peptide, cathelicidin 4 (CATHL4), cathelicidin 6 (CATHL6) have been reported in other species as milk production related gene. For this reason, we concluded that our detected 271 genes would be strong candidates for determining milk production.

Studies on Seeding Mechanized Culture of Wheat in Paddy Field (답리작 소맥의 파종 기계화 재배에 관한 연구)

  • 박문수;장민식;이강세;박노풍
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.24 no.3
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    • pp.35-42
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    • 1979
  • Experiments were conducted to find out the labor saving efficiency and optimum seeding rate and fertilizer level when newly invented Barley Rotor Caster was used in wheat sowing on paddy field. It decreased the labor for sowing by 66~77% compared with conventional sowing method. And the optimum. seeding rate was considered to be approximately 20$\ell$Q per 10a in standard fertilizer level and optimum fertilizer level to be 18-12-12kg/10a, $N-P_2O_5-K_2O$ respectively.

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Calcium Channel Blockers Suppress the Responses of Rat Dorsal Horn Cell to Nociceptive Input (쥐 척수후각세포의 유해자극 반응에 대한 칼슘이온통로 차단제의 억제작용)

  • Kang, Sok-Han;Kim, Kee-Soon;Shin, Hong-Kee
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.6
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    • pp.625-637
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    • 1997
  • Calcium ions are implicated in a variety of physiological functions, including enzyme activity, membrane excitability, neurotransmitter release, and synaptic transmission, etc. Calcium antagonists have been known to be effective for the treatment of exertional angina and essential hypertension. Selective and nonselective voltage-dependent calcium channel blockers also have inhibitory action on the acute and tonic pain behaviors resulting from thermal stimulation, subcutaneous formalin injection and nerve injury. This study was undertaken to investigate the effects of iontophoretically applied $Ca^{++}$ and its antagonists on the responses of WDR (wide dynamic range) cells to sensory inputs. The responses of WDR cells to graded electrical stimulation of the afferent nerve and also to thermal stimulation of the receptive field were recorded before and after iontophoretical application of $Ca^{++}$, EGTA, $Mn^{++}$, verapamil, ${\omega}-conotoxin$ GVIA, ${\omega}-conotoxin$ MVIIC and ${\omega}-agatoxin$ IVA. Also studied were the effects of a few calcium antagonists on the C-fiber responses of WDR cells sensitized by subcutaneous injection of mustard oil (10%). Calcium ions and calcium channel antagonists ($Mn^{++}$, verapamil, ${\omega}-conotoxin$ GVIA & ${\omega}-agatoxin$ IVA) current-dependently suppressed the C-fiber responses of WDR cells without any significant effects on the A-fiber responses. But ${\omega}-conotoxin$ MVIIC did not have any inhibitory actions on the responses of WDR cell to A-fiber, C-fiber and thermal stimulation. Iontophoretically applied EGTA augmented the WDR cell responses to C-fiber and thermal stimulations while spinal application of EGTA for about $20{\sim}30\;min$ strongly inhibited the C-fiber responses. The augmenting and the inhibitory actions of EGTA were blocked by calcium ions. The WDR cell responses to thermal stimulation of the receptive field were reduced by iontophoretical application of $Ca^{++}$, verapamil, ${\omega}-agatoxin$ IVA, and ${\omega}-conotoxin$ GVIA but not by ${\omega}-conotoxin$ MVIIC. The responses of WDR cells to C-fiber stimulation were augmented after subcutaneous injection of mustard oil (10%, 0.15 ml) into the receptive field and these sensitized C-fiber responses were strongly suppressed by iontophoretically applied $Ca^{++}$, verapamil, ${\omega}-conotoxin$ GVIA and ${\omega}-agatoxin$ IVA. These experimental findings suggest that in the rat spinal cord, L-, N-, and P-type, but not Q-type, voltage-sensitive calcium channels are implicated in the calcium antagonist-induced inhibition of the normal and the sensitized responses of WDR cells to C-fiber and thermal stimulation, and that the suppressive effect of calcium and augmenting action of EGTA on WDR cell responses are due to changes in excitability of the cell.

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Biosurfactant as a microbial pesticide

  • Lee, Baek-Seok;Choi, Sung-Won;Choi, Ki-Hyun;Lee, Jae-Ho;Kim, Eun-Ki
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.40-44
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    • 2003
  • Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

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Vegetational Management Planning of the Namsan Nature Park in Seoul on the Basis of Phytosociological Characteristics (식물사회학적 특성을 고려한 남산공원 식생의 관리대책)

  • Oh, Koo Kyoon;Lee, Kyong Jae;Yim, Kyong Bin
    • Journal of Korean Society of Forest Science
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    • v.77 no.1
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    • pp.1-9
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    • 1988
  • To analyze phytosociological characteristics of the Mt. Namsan nature park, Seoul, vegetation structure of north-east and south-west slopes, competition capability among species, vegetation structures of afforested species were studied with belt-transect and quadrat methods. And management plan of each vegetational zone was proposed by the results of this study and the other previous reports. Numbers of species and individuals of south-west slope were greater than those of north-east slope. But species diversity of northeast showed stability of 1.0, while that of south-west showed instability from 0.8 to 1.3. The floristic dissimilarity coefficient between two slopes showed 70-80% in which species composition between two slopes was much different. Quercus mongolica was a dominant species at north-east slope and Pinus densiflora at south-west slope. Competition capability among major species was resulted in deciduous trees>Robinia pseudoacacia>Populus alba ${\times}$ glandulosa>Pinais densiflora, Quercus mongolica>deciduous trees>Pinus densiflora. Afforested vegetation at the Mt. Namsan, especially Pinus koraiensis, Abies holophylla, Picea abies, etc showed poor growth rate. The goal of the Nam-san nature park management is to conserve natural ecosystem as it is and, if necessary, to accerlerate restoration of the disturbed ecosytem to its nature by minimum energy input as possible. Management proposal was made for six vegetational management zone ; P. derasitZoru forest zone, Q. mongolica forest zone, deciduous forest zone, afforestation zone, R. pseduoacacia zone and landscape planting zone.

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Installation and Data Analysis of Superconducting Gravimeter in MunGyung, Korea; Preliminary Results (문경 초전도 중력계 설치 및 기초자료 분석)

  • Kim, Tae-Hee;Neumeyer, Juergen;Woo, Ik;Park, Hyuck-Jin;Kim, Jeong-Woo
    • Economic and Environmental Geology
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    • v.40 no.4
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    • pp.445-459
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    • 2007
  • Superconducting Gravimeter(SG) was installed and has been successfully operated at MunGyung, Kyungsang province in Korea in March 2005. It was registered as the 21st observatory of the Global Geodynamics Project. Since SG can precisely measure the gravity variations below the 1mHz frequency band, it has the outstanding capability to sense and resolve many different periodic gravity components from each other. From the raw data collected between 18 March 2005 and 21 February 2006 diurnal and semi-diurnal tidal band's residual gravity components were analyzed. During this process, the instrumental noises, air pressure, and ground water corrections were carried out. Values of $-3.18nm/s^2/hPa\;and\;17nm/s^2/m$ were used respectively in the air pressure and groundwater corrections. Hartmann-Wenzel and Whar-Dehant Earth tide models were adopted to compute the residual gravity for Q1, O1, P1, K1, M2, N2, S2, K2 tidal bands. For the ocean loading correction, SCW80, FES952, and FES02 models were used and compared. As a result, FES02 ocean loading model has shown the best match for the data processing at MunGyung SG MunGyung SG gravity was compared with GRACE satellite gravity. The correlation coefficient between the two gravity after groundwater correction was 0.628, which is higher than before ground water correction. To evaluate sensitivity at MunGyung SG gravity statition, the gravity data measured during 2005 Indodesian earthquake was compared with STS-2 broad band seismometer data. The result clearly revealed that the SG could recorded the same period of earthquake with seismometer event and a few after-shock events those were detected by seismometer.