• Title/Summary/Keyword: strain selection

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Molecular Mechanism of ABC Transporter Mdr49A Associated with a Positive Cross-Resistance in Transgenic Drosophila (형질전환 초파리를 이용한 Mdr49A 유전자의 살충제 교차저항성 기능 구명)

  • Seong, Keon Mook;Pittendrigh, Barry R.
    • Korean journal of applied entomology
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    • v.59 no.4
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    • pp.341-348
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    • 2020
  • The ATP-binding cassette (ABC) transporter superfamily represents the largest transmembrane protein that transports a variety of substrates across extra- and intra-cellular membranes. In insects, the ABC transporter proteins play crucial roles in insecticide resistance. To date, no studies have investigated the involvement of ABC transporter gene for cross-resistance to insecticide chemistries. Here, we studied such possible mechanisms against six conventional insecticides using transgenic Drosophila melanogaster strains carrying Mdr49 transcript variant A. For the 91-R and 91-C strains of Drosophila melanogaster, although they have a common origin, 91-R has been intensely selected with DDT for over 60 years, while 91-C has received no insecticide selection. Our transgenic analyses showed that overexpression of 91-R-MDR49 transcript variant A along with three amino acid variations can yield a relatively low degree of cross-resistance to carbofuran (2.0~6.7-fold) and permethrin (2.5~10.5-fold) but did not show cross-resistance to abamectin, imidacloprid, methoxychlor, and prothiofos as compared to the Gal4-driver control strain without transgene expression. These results indicate that the overexpression of Mdr49A in itself leads to a cross-resistance and three amino acid changes have additional effects on positive cross-resistance to carbofuran and permethrin.

Evaluation of Acaricidal Effect Against Two-spotted Spider Mite Collected from Strawberry in Greenhouse (딸기 시설 하우스 점박이응애에 대한 살비 효과 평가)

  • Kwon, D.H.;Ahn, Y.K.;Hong, K.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.22 no.2
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    • pp.135-146
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    • 2020
  • In the strawberry cultivation areas and domestic farms, two-spotted spider mites are the most serious pests. It decreases the product yield due to the direct feeding of chlorophyll by the mites resulting in reduced photosynthetic ability of host plant. In this study, a simplified acaricidal effect evaluation system (SAEES) was employed to choose the effective acaricidal products among 10 items based on leaf-dipping bioassay methods. SAEES had the advantage of being able to screen four commercial products with three replications at the same time in the recommended concentration. The susceptible strains (SL_YS) showed a high mortality rate of about 90-100%, whereas the mortality of local strains was differed by each acaricides. It suggests that the acaricide responses of field populations might differ due to spray frequencies and acaricide product. An efficacy index (0.8-1.0) was determined based on the mortality of susceptible strain, which would allow the most effective commercial products to be selected by the range of this index. In summary, SAEES will enable the selection of effective commercial products and contribute to increasing control against T. urticae in strawberries.

Optimization of Protoplast Isolation and Ribonucleoprotein/Nanoparticle Complex Formation in Lentinula edodes (표고버섯의 원형질체 분리 최적화와 RNPs/나노파티클 복합체 형성)

  • Kim, Minseek;Ryu, Hojin;Oh, Min Ji;Im, Ji-Hoon;Lee, Jong-Won;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.20 no.3
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    • pp.178-182
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    • 2022
  • Despite the long history of mushroom use, studies examining the genetic function of mushrooms and the development of new varieties via bio-molecular methods are significantly lacking compared to those examining other organisms. However, owing to recent developments, attempts have been made to use a novel gene-editing technique involving CRISPR/Cas9 technology and genetic scissors in mushroom studies. In particular, research is actively being conducted to utilize ribonucleoprotein particles (RNPs) that can be genetically edited with high efficiency without foreign gene insertion for ease of selection. However, RNPs are too large for Cas9 protein to pass through the cell membrane of the protoplasmic reticulum. Furthermore, guide RNA is unstable and can be easily decomposed, which remarkably affects gene editing efficiency. In this study, nanoparticles were used to mitigate the shortcomings of RNP-based gene editing techniques and to obtain transformants stably. We used Lentinula edodes (shiitake mushroom) Sanjo705-13 monokaryon strain, which has been successfully used in previous genome editing experiments. To identify a suitable osmotic buffer for the isolation of protoplast, 0.6 M and 1.2 M sucrose, mannitol, sorbitol, and KCl were treated, respectively. In addition, with various nanoparticle-forming materials, experiments were conducted to confirm genome editing efficiency via the formation of nanoparticles with calcium phosphate (CaP), which can be bound to Cas9 protein without any additional amino acid modification. RNPs/NP complex was successfully formed and protected nuclease activity with nucleotide sequence specificity.

Current Status and Prospects for Standards, Regulations, and Detection of Probiotic Yogurt: Review (프로바이오틱 요구르트의 기준, 규정, 검출에 관한 현황 및 전망: 총설)

  • Jung-Whan Chon;Kun-Ho Seo;Tae-Jin Kim;Hye-Young Youn;Seok-Hyeong Kang;Won-Uk Hwang;Hajeong Jeong;Dongkwan Jeong;Kwang-Young Song
    • Journal of Dairy Science and Biotechnology
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    • v.41 no.1
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    • pp.9-25
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    • 2023
  • Yogurt fermentation is known to be beneficial because it provides a low pH and harsh environment for foodborne pathogens and improves organoleptic properties. Additionally, organic acids produced through fermentation have a good effect on the viscosity and gelling properties of yogurt. Several potential health benefits of probiotic and generally recognized as safe strains have been suggested. Yogurt is the preferred vehicle for delivering probiotics to health-conscious consumers. Therefore, manufacturers of probiotic beverages must comply with the relevant regulations. The development of probiotic yogurt begins with the selection of strains with safety and functional properties of probiotics. The selected probiotic strain should be technically suitable for viability and improve organoleptic quality while maintaining the number of bacteria above the standard value during processing and storage conditions. In addition, the efficacy of probiotic strains contained in yogurt should be investigated, confirmed, and approved according to well-designed clinical trials. Although various methods are used to detect probiotic strains, the recently widely used next generation sequencing method can be actively utilized. In the future, more research should be conducted with the latest methods to identify probiotic functions and accurately detect probiotic strains.

Introduction, Development, and Characterization of Supernodulating Soybean Mutant. 1. Mutagenesis of Soybean and Selection of Supernodulating Soybean Mutant (다량 뿌리혹 형성 콩 계통의 도입 개발 및 생육특성구명 1. 돌연변이유기에 의한 콩 초다뿌리혹형성 계통선발)

  • Lee, Hong-Suk;Chae, Young-Am;Park, Eui-Ho;Kim, Yong-Wook;Yun, Kwang-Il;Lee, Suk-Ha
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.2
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    • pp.247-253
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    • 1997
  • Development of soybean cultivars with great nodulation and high nitrogen fixation activity, derived mostly from mutagenesis, may decrease inputs of chemical fertilizer nitrogen into the soil-plant system. Soybean seeds (cv. Jangyupkong, Hwanggeumkong, and Geomjungkong 1) were treated with three different levels of EMS (ethyl methanesulfonate) concentration(30, 50, and 70mM). Increasing the doses of EMS resulted in decreased field emergence rate of seeds, whereas it did not increase M$_2$ mutation frequencies. This indicated that the most efficient concentration of EMS was 30mM for generating mutants. Extensive mutagenesis of Sinpaldalkong 2 with 30mM EMS was undertaken to isolate soybean mutants with greater nodulation. Approximately 8, 200 M$_2$ families were screened for greater nodulation on 5 mM nitrate after inoculation with Bradyrhizobium japonicum strain YCK213-KFCC-10728. Mutant SS-2 nodulated more than the wild type. Comparison of supernodulation between SS-2 and two nts mutants(nts 1007 and nts 1116) revealed that SS-2 showed the supernodulation character at an earlier growth stage than the two nts mutants. Further studies should be needed to characterize the difference in timing of nodulation between SS-2 and nts mutants.

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Conserved COG Pathways and Genes of 122 Species of Archaea (고세균 122종의 보존적 COG pathways와 유전자)

  • Dong-Geun Lee ;Sang-Hyeon Lee
    • Journal of Life Science
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    • v.33 no.11
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    • pp.944-949
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    • 2023
  • The purpose of this study was to identify conserved metabolic pathways and conserved genes in 122 archaeal species. Using the Clusters of Orthologous Groups of Proteins (COG) database of conserved genes, we analyzed whether 122 species had 63 COG metabolic pathways, the 822 COGs that compose them, and a total of 4,877 COGs. Archaeal ribosomal proteins were the most conserved in metabolic pathways. 46 COGs in seven COG pathways among 63 COG pathways and 20 COGs in others were conserved in 122 species. Some genes involved in cell wall and extracellular matrix synthesis, replication, transcription, translation, and protein metabolism were common to all 122 species. When the distance value of the phylogenetic tree was analyzed at the phylum level or class level, the average was the lowest at the class Halobacteria of the phylum Euryarchaeota. Standard deviation was high for the class Nitosospharia of the phylum Thaumarchaeota, the unclassified members of phylum Thaumarchaeota, the class Halobacteria of the phylum Euryarchaeota, the class Thermoprotei of the phylum Crenarchaeota, and other archaea. Furthermore, the phylogenetic tree analysis revealed six commonalities. The results of this study, along with data on conserved genes, could be used for drug development and gene selection for strain improvement.

FT-transgenic spray-type Chrysanthemum (Dendranthema grandiflorum Kitamura) showing early-flowering (FT 유전자 형질전환 스프레이 국화 (Dendranthema grandiflorum (Ramat.) Kitamura)의 조기개화성)

  • Lee, Su-Young;Han, Bong-Hee;Hur, Eun-Joo;Shin, Hak-Kee;Lee, Il-Ha;Lee, Eun-Kyung;Kim, Seung-Tae;Kim, Won-Hee;Kwon, O-Hyeon
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.140-145
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    • 2012
  • The flowering locus T (FT) gene, of which expression will be controlled at high temperature by heat shock promoter (it printed as to HSproFT), was introduced into spray-type chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) 2 cultivars ('Pink PangPang' and 'Pink Pride' by co-cultivation with Agrobacterium tumefaciens strain C58C1 harboring pCAMBIA2300 containing the HSproFT gene. After leaf segments of the 2 cultivars were infected with the A. tumafaciens with C58C1 as explants, shoots were regenerated from the explants cultured on the $1^{st}$ selection medium (MS basal salts + 1.0 mg/L BA, 0.5 mg/L IAA + 10 mg/L kanamycin + 0.7% plant agar, pH 5.8). The shoots were transferred into the $2^{nd}$ selection medium (MS basal salts + 1.0 mg/L BA, 0.5 mg/L IAA + 20 mg/L kanamycin + 0.7% plant agar, pH 5.8). One hundred seventeen plantlets from 'Pink PangPang' and 5 ones from 'Pink Pride' were confirmed as transformants by PCR analysis. Twenty six of the transformants and non-transformants were acclimatized and established well in a green house. Eights of 26 transgenic lines showed flower bud 1.7~10 days earlier than nontransgenic plants, and 24 of them flowered 1~6 days earlier than non- transgenic plants. The shape and color of flower of all HSproFT-transgenic lines were not different with those of non- transgenic plants.

Recurrent parent genome (RPG) recovery analysis in a marker-assisted backcross breeding based on the genotyping-by-sequencing in tomato (Solanum lycopersicum L.) (토마토 MABC 육종에서 GBS(genotyping-by-sequencing)에 의한 RPG(recurrent parent genome) 회복률 분석)

  • Kim, Jong Hee;Jung, Yu Jin;Seo, Hoon Kyo;Kim, Myong-Kwon;Nou, Ill-Sup;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.165-171
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    • 2019
  • Marker-assisted backcrossing (MABC) is useful for selecting an offspring with a highly recovered genetic background for a recurrent parent at early generation to various crops. Moreover, marker-assisted backcrossing (MABC) along with marker-assisted selection (MAS) contributes immensely to overcome the main limitation of the conventional breeding and it accelerates recurrent parent genome (RPG) recovery. In this study, we were employed to incorporate rin gene(s) from the donor parent T13-1084, into the genetic background of HK13-1151, a popular high-yielding tomato elite inbred line that is a pink color fruit, in order to develop a rin HK13-1084 improved line. The recurrent parent genome recovery was analyzed in early generations of backcrossing using SNP markers obtained from genotyping-by-sequencing analysis. From the $BC_1F_1$ and $BC_2F_1$ plants, 3,086 and 4868 polymorphic SNP markers were obtained via GBS analysis, respectively. These markers were present in all twelve chromosomes. The background analysis revealed that the extent of RPG recovery ranged from 56.7% to 84.5% and from 87.8% to 97.8% in $BC_1F_1$ and $BC_2F_1$ generations, respectively. In this study, No 5-1 with 97.8% RPG recovery rate among $BC_2F_1$ plants was similar to HK13-1151 strain in the fruit shape. Therefore, the selected plants were fixed in $BC_2F_2$ generation through selfing. MAS allowed identification of the plants that are more similar to the recurrent parent for the loci evaluated in the backcross generations. MABC can greatly reduce breeding time as compared to the conventional backcross breeding. For instance, MABC approach greatly shortened breeding time in tomato.

Characteristics and pedigree selection of a shortened cultivation period strain in Lepista nuda (재배기간이 짧은 민자주방망이버섯 우량계통 선발 및 특성)

  • Jeon, Jong-Ock;Lee, Kwan-Woo;Lee, Kyoung-Jun;Kim, Min-Ja;Kim, In-Jae;Kim, Young-Ho
    • Journal of Mushroom
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    • v.18 no.4
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    • pp.331-338
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    • 2020
  • This study was conducted to cultivate new Lepista nuda varieties with shorter cultivation period and better fruiting body compared to that of wild strains, for mass production and commercial application. Eighteen genetic resources of L. nuda were collected and grown in boxes using rice straw-fermented growth medium. Four lines with fruiting bodies were formed and selected as cross-breeding lines. Although 657 combinations were crossed through monospore crossing, only 17 combinations were bred between the 'CBMLN-19' line and the 'CBMLN-30' line. Among them, 8 lines with fast mycelial growth and high density were selected. After inoculating the rice straw-fermented growth medium with 14 genetic resources and 8 cross-breeding lines, their incubation period was investigated. Six of the cross-breeding lines completed their incubation in 20 days, while 7 of the 14 genetic resources took more than 40 days to complete their incubation, reducing the incubation period by more than 20 days in most cross-breeding lines. After the incubations were completed, the clay loam soil was covered with for post-cultivation, and when the mycelial cultivation was complete, the formation of fruiting bodies was induced after scraping the mycelial bodies under these environmental conditions: 14℃, 95% relative humidity or higher, and 1,500 to 2,000 ppm CO2 concentration. The temperature was reduced to 6℃ at night, resulting in a low temperature shock. Thus, 4 lines of fruiting bodies occurred from two genetic resources 'CBMLN-31' and 'CBMLN-44' and two cross-bred lines 'CBMLN-96' and 'CBMLN-103'. After inoculation, the longest period for fruiting bodies to occur was 100 days for the control:, the genetic resource 'CBMLN-31', and the shortest period (45 days) was observed for the cross-breeding line 'CBMLN-103'. The result of the investigation of the fruiting body characteristics shows that the cross-bred line 'CBMLN-103' showed a small form with 1.9 g of individual weight and 123validstipes per box, which was the highest incidence among the four lines. Another cross-bred line, 'CBMLN-96', had an individual weight of 5.5 g, which is larger than that of 'CBMLN-103'; however, the number of valid stipes per box was 30 less than that of 'CBMLN-103'. Quantity analysis showed that the control, 'CBMLN-31', had the highest quantity of 783 g per box, followed by the cross-bred line, 'CBMLN-96' with 165 g per box, and then the 'CBMLN-103' with 232 g. The quantity of the two crossbred lines was lower than that of the control 'CBMLN-31'; however, the amount of fruiting bodies was higher, and the cultivation period was shortened by 32 to 33 days. Therefore, these two lines would be selected as superior lines.

Serological Approach for Selection of Bradyrhizobium japonicum Strain with Superior Symbiotic Effectiveness (Bradyrhizobium japonicum의 공생효과(共生效果) 우수균주(優秀菌株) 선발(選拔)을 위한 면역혈청학적(免疫血淸學的) 접근(接近))

  • Kang, Ui-Gum;Ha, Ho-Sung;Park, Kyeong-Bae;Lee, Sang-Kyu;Lim, Dong-Kyu;Yang, Min-Suk
    • Korean Journal of Soil Science and Fertilizer
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    • v.29 no.2
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    • pp.165-172
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    • 1996
  • Symbiotic effectiveness and significance of Bradyrhizobium japonicum strains in five serogroups which were consisted of three corresponding serotype strains, respectively, indigenous to Korean soils were evaluated in terms of utility of strain serogroup for symbiotic improvement on soybean plant. The nodulation by rhizobia of each serogroup on four soybean cultivars(Danweonkong, Kwangkyo, Pangsakong, and Eunhakong) was good in order of USDA 123 > YCK 150 > YCK 117 > YCK 141 > USDA 110 serogroup members. Shoot dry weight of soybean was relatively high with USDA 110 serogroup members as well as with YCK 141 serogroup members, whereas the effectiveness of USDA 123 serogroup members was the lowest among the serogroups examined. In particular, Pangsakong among soybeans inoculated with five-serogroup members was positively outstanding on nodulation and shoot dry weight of the plant. Overall, symbiotic parameters of serogroup members associated with soybean plant such as nodule number, nodule mass, $N_2$ase activity, and shoot dry weight showed significantly different responses at level of 1% probability among both rhizobial serogroups and soybean cultivars, respectively. The rate in symbiotic similarity of the members of each serogroup from F-test ($$P{\leq_-}0.05$$) was 100% for nodule No., 90% for $N_2$ase activity. and 80% for soybean shoot dry weight. Taken together, the results indicated that the serological grouping of B. japonicum could be strongly useful for improving the symbiotic effectiveness hetween soybean and Rhizobium.

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