Development of In Vitro Seedling Screening Method for Selection of Resistant Rice Against Bakanae Disease

ABSTRACT We have constructed an improved recombination-based in vivo expression technology (RIVET) and used it as a screening method to identify Vibrio cholerae genes that are transcriptionally induced during infection of infant mice. The improvements include the introduction of modified substrate cassettes for resolvase that can be positively and negatively selected for, allowing selection of resolved strains from intestinal homogenates, and three different tnpR alleles that cover a range of translation initiation efficiencies, allowing identification of infection-induced genes that have low-to-moderate basal levels of transcription during growth in vitro. A transcriptional fusion library of 8,734 isolates of a V. cholerae El Tor strain that remain unresolved when the vibrios are grown in vitro was passed through infant mice, and 40 infection-induced genes were identified. Nine of these genes were inactivated by in-frame deletions, and their roles in growth in vitro and fitness during infection were measured by competition assays. Four mutant strains were attenuated >10-fold in vivo compared with the parental strain, demonstrating that infection-induced genes are enriched in genes essential for virulence.

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Development of an Efficient In Vitro Screening Method for Selection of Resistant Lily Cultivars Against Fusarium oxysporum f. sp. lilii

Korean Journal of Horticultural Science and Technology ◽

10.7235/hort.2015.15085 ◽

2015 ◽

Vol 33 (6) ◽

pp. 883-890 ◽

Cited By ~ 1

Author(s):

Ji-Young Jang ◽

Ki-Beom Moon ◽

Jang-Ho Ha ◽

Ji-Sun Park ◽

Mi-Jin Kim ◽

...

Keyword(s):

Fusarium Oxysporum ◽

Screening Method ◽

In Vitro Screening ◽

Selection Of

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High-throughput screening of biomolecules using cell-free gene expression systems

Synthetic Biology ◽

10.1093/synbio/ysy012 ◽

2018 ◽

Vol 3 (1) ◽

Cited By ~ 15

Author(s):

Luis E Contreras-Llano ◽

Cheemeng Tan

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Screening Method ◽

Screening Methods ◽

Free Gene ◽

On Chip ◽

Translation Systems ◽

Selection Of

Abstract The incorporation of cell-free transcription and translation systems into high-throughput screening applications enables the in situ and on-demand expression of peptides and proteins. Coupled with modern microfluidic technology, the cell-free methods allow the screening, directed evolution and selection of desired biomolecules in minimal volumes within a short timescale. Cell-free high-throughput screening applications are classified broadly into in vitro display and on-chip technologies. In this review, we outline the development of cell-free high-throughput screening methods. We further discuss operating principles and representative applications of each screening method. The cell-free high-throughput screening methods may be advanced by the future development of new cell-free systems, miniaturization approaches, and automation technologies.

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Efficient In Vitro Screening for Higher Soil pH Adaptability of Intersectional Hybrids in Blueberry

HortScience ◽

10.21273/hortsci.49.2.141 ◽

2014 ◽

Vol 49 (2) ◽

pp. 141-144 ◽

Cited By ~ 5

Author(s):

Hirotoshi Tsuda ◽

Hisato Kunitake ◽

Yo Aoki ◽

Akiko Oyama ◽

Takuya Tetsumura ◽

...

Keyword(s):

Screening Method ◽

Multiple Shoots ◽

Vaccinium Corymbosum ◽

Apparent Difference ◽

In Vitro Screening ◽

Ph Tolerance ◽

Short Time ◽

Hybrid Clones ◽

Selection Of

We tested efficient in vitro methods for screening the genotypes with higher pH tolerance using multiple shoots of intersectional hybrids between Vaccinium corymbosum ‘Spartan’ and V. bracteatum. The response of the four hybrid clones tested to different pH levels was clone-dependent in vitro. An apparent difference was found in the rooting rate among the hybrid clones even at higher pH levels; the rooting rates of JM4 (91%) at pH 8.0 indicated a significantly high value compared with other clones (JM1: 24%, JM2: 9%, JM3: 8%, ‘Spartan’: 0%). Furthermore, JM4 showed constantly high rooting rates (91% to 100%) at all pH levels with no significant differences. Similar differences in the root characters of the hybrids were also confirmed by checking the viability of roots using fluorescein diacetate (FDA)/propidium iodide (PI) staining after dipping the roots of in vitro-produced shoots in liquid medium at different pH levels for 6 hours. These results suggest that an in vitro screening method using the rooting rate of multiple shoots and the viability test of roots by FDA/PI staining as a marker could become a very useful tool for the selection of germplasm with tolerance to higher pH within a short time using small planting spaces. In addition, JM4, which showed a high rooting rate at pH 8.0, could be useful in breeding new cultivars with higher pH tolerance.

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The effect of fungicides used in the protection of forest tree seedlings on the growth of ectomycorrhizal fungi

Acta Mycologica ◽

10.5586/am.1997.028 ◽

2014 ◽

Vol 32 (2) ◽

pp. 315-322 ◽

Cited By ~ 3

Author(s):

Marta Aleksandrowicz-Trzcińska ◽

Andrzej Grzywacz

Keyword(s):

Ectomycorrhizal Fungi ◽

Screening Method ◽

Forest Tree ◽

Tree Seedlings ◽

Laccaria Laccata ◽

Suillus Luteus ◽

Study Results ◽

Forest Nurseries ◽

Selection Of

Fungitoxical activity of ten fungictdes most commonly used in the phytopathological protection of forest nurseries was studied, using the in vitro screening method. The fungitoxical activity was studied against five species of ectomycorrhizal fungi (seven strains). The resulting growth inhibition of fungi species and strains tested was prcscnted in terms of fungitoxicity classes of the preparations used. The highest total fungitoxicity against the mycelia of fungi taxa tested was found for Euparen, Bravo, Dithane M-45 and Ridomil. The weakest fungitoxical effect was observed for Topsin M and Bayleton. The least susceptible for the action of the fungicides studied were mycelia of Suillus luteus, while the most susceptible were those of Hebeloma crustuliniforme and Laccaria laccata. The study results arę useful for the selection of fungi strains proper for the artificial mycorrhization of seedlings.

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In vitro technique for detecting tegument damage in Diclidophora merlangi: Possible screening method for selection of undamaged tissues or organisms prior to physiological investigation

Experimental Parasitology ◽

10.1016/0014-4894(71)90069-5 ◽

1971 ◽

Vol 30 (1) ◽

pp. 54-57 ◽

Cited By ~ 29

Author(s):

D.W. Halton ◽

C. Arme

Keyword(s):

Screening Method ◽

Physiological Investigation ◽

In Vitro Technique ◽

Selection Of

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An integrated screening method for evaluating the pulmonary toxicity of inhaled particulates: Role of microscopic techniques in assessing pulmonary macrophage clearance functions

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100161692 ◽

1990 ◽

Vol 48 (3) ◽

pp. 826-827

Author(s):

David B. Warheit ◽

Lena Achinko ◽

Mark A. Hartsky

Keyword(s):

Bronchoalveolar Lavage ◽

Pulmonary Toxicity ◽

Screening Method ◽

Pulmonary Response ◽

Inhaled Particles ◽

Cytochemical Analysis ◽

Pulmonary Macrophages ◽

Integrated Screening

There is a great need for the development of a rapid and reliable bioassay to evaluate the pulmonary toxicity of inhaled particles. A number of methods have been proposed, including lung clearance studies, bronchoalveolar lavage analysis, and in vitro cytotoxicity tests. These methods are often limited in scope inasmuch as they measure only one dimension of the pulmonary response to inhaled, instilled or incubated dusts. Accordingly, a comprehensive approach to lung toxicity studies has been developed.To validate the method, rats were exposed for 6 hours or 3 days to various concentrations of either aerosolized alpha quartz silica (Si) or carbonyl iron (CI) particles. Cells and fluids from groups of sham and dust-exposed animals were recovered by bronchoalveolar lavage (BAL). Alkaline phosphatase, LDH and protein values were measured in BAL fluids at several time points postexposure. Cells were counted and evaluated for viability, as well as differential and cytochemical analysis. In addition, pulmonary macrophages (PM) were cultured and studied for morphology, chemotaxis, and phagocytosis by scanning electron microscopy.

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