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

HortScience ◽  
2014 ◽  
Vol 49 (2) ◽  
pp. 141-144 ◽  
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.

Development of an in vitro screening method for safety evaluation of nanomaterials

2009 ◽  
Vol 19 (1) ◽  
pp. 19-27 ◽  
Author(s):  
Atsuko Matsuoka ◽  
Agneta Önfelt ◽  
Yoshie Matsuda ◽  
Ryusuke Nakaoka ◽  
Yuji Haishima ◽  
...  
Keyword(s):  
Screening Method ◽  
Safety Evaluation ◽  
In Vitro Screening

AN IN-VITRO SCREENING METHOD TO STUDY THE ACTIVITY POTENTIAL OF BIOFERTILIZERS BASED ONTRICHODERMAANDBACILLUSSP.

2013 ◽  
Vol 36 (9) ◽  
pp. 1439-1452 ◽  
Author(s):  
Zafrin Akter ◽  
Markus Weinmann ◽  
Günter Neumann ◽  
Volker Römheld
Keyword(s):  
Screening Method ◽  
In Vitro Screening

scholarly journals Second-Generation Recombination-Based In Vivo Expression Technology for Large-Scale Screening for Vibrio cholerae Genes Induced during Infection of the Mouse Small Intestine

2005 ◽  
Vol 73 (2) ◽  
pp. 972-980 ◽  
Author(s):  
C. G. Osorio ◽  
J. A. Crawford ◽  
J. Michalski ◽  
H. Martinez-Wilson ◽  
J. B. Kaper ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Large Scale ◽  
Screening Method ◽  
Mouse Small Intestine ◽  
In Vivo Expression ◽  
Induced Genes ◽  
El Tor ◽  
Selection Of

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.

scholarly journals 057 IN VITRO SCREENING OF WESTERN UNITED STATES VACCINIUM SPECIES FOR pH TOLERANCE

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 436b-436
Author(s):  
Amy J. Moberg ◽  
James J. Luby ◽  
Carl J. Rosen ◽  
Peter D. Ascher
Keyword(s):  
United States ◽  
Succinic Acid ◽  
Root Zone ◽  
Western United States ◽  
Screening Procedure ◽  
In Vitro Screening ◽  
Essential Nutrients ◽  
Ph Tolerance

Accessions of Vaccinium species (deliciosum, ovalifolium, membranaceum, parvifolium, scoparium) were evaluated for tolerance to higher pH in the root zone using an in vitro screening procedure. Seeds were germinated on media containing all essential nutrients with nitrogen in the nitrate form at pH 5 and pH 6 and evaluated for 21 weeks. Excess EDTA was used to buffer the micronutrients and pH was buffered by MES and succinic acid. Germination varied among species with V. ovalifolium being highest and V. parvifolium not germinating at all. Mortality was lower at pH 5. At pH 6, V. ovalifolium and V. membranaceum exhibited variation for growth while all other species suffered complete mortality.

scholarly journals Evaluation in Vitro of Blueberry Germplasm for Higher pH Tolerance

1991 ◽  
Vol 116 (2) ◽  
pp. 312-316 ◽  
Author(s):  
Chad E. Finn ◽  
James J. Luby ◽  
Carl J. Rosen ◽  
Peter D. Ascher
Keyword(s):  
Combining Ability ◽  
Variance Components ◽  
Growth Traits ◽  
Dry Weight ◽  
Vaccinium Corymbosum ◽  
High Ph ◽  
Ph Tolerance ◽  
Radicle Emergence ◽  
Ethanesulfonic Acid

Progenies from crosses among eight highbush (Vaccinium corymbosum L.), lowbush (V. angustifolium Ait.), and V. corymbosum/V. angustifolium hybrid-derivative parents were evaluated in vitro at low (5.0) and high (6.0) pH for vitality, height, and dry weight. Succinic acid and 2[N- morpholino]ethanesulfonic acid (Mes) effectively maintained pH in the medium and rhizosphere. The pH regime did not affect percent radicle emergence from seed or survival; however, percent seed germination was slightly lower at high pH. The parental general combining ability (GCA), reciprocal and maternal, but not the specific combining ability (SCA) variance components were significant for plant vitality, height, and dry weight. The GCA variance components were six to 26 times larger than the SCA variance components for the plant growth traits. Variation due to pH regime was significant for vitality and dry weight but not for plant height. The progenies of parents with high percent lowbush ancestry were taller at both pH levels than those with less such ancestry. Little variation was apparent for higher pH tolerance as measured by dry weight; however, the GCA effects suggested that the progenies of some parents performed better than others at high pH. Vaccinium angustifolium parents differed in the extent to which tolerance to high pH was transmitted. In vitro screening in concert with a traditional breeding program should be effective in improving blueberry tolerance to higher pH.

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

2018 ◽  
Vol 3 (1) ◽  
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.

scholarly journals Simple rapid in vitro screening method for SARS-CoV-2 anti-virals that identifies potential cytomorbidity-associated false positives

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Kexin Yan ◽  
Daniel J. Rawle ◽  
Thuy T. Le ◽  
Andreas Suhrbier
Keyword(s):  
Cell Density ◽  
Drug Screening ◽  
Screening Method ◽  
False Positives ◽  
In Vitro Screening ◽  
Viral Activity ◽  
Drug Concentrations ◽  
In Vitro Drug Screening ◽  
Growth Assay

Abstract Background The international SARS-CoV-2 pandemic has resulted in an urgent need to identify new anti-viral drugs for treatment of COVID-19. The initial step to identifying potential candidates usually involves in vitro screening that includes standard cytotoxicity controls. Under-appreciated is that viable, but stressed or otherwise compromised cells, can also have a reduced capacity to replicate virus. A refinement proposed herein for in vitro drug screening thus includes a simple growth assay to identify drug concentrations that cause cellular stress or “cytomorbidity”, as distinct from cytotoxicity or loss of viability. Methods A simple rapid bioassay is presented for antiviral drug screening using Vero E6 cells and inhibition of SARS-CoV-2 induced cytopathic effects (CPE) measured using crystal violet staining. We use high cell density for cytotoxicity assays, and low cell density for cytomorbidity assays. Results The assay clearly illustrated the anti-viral activity of remdesivir, a drug known to inhibit SARS-CoV-2 replication. In contrast, nitazoxanide, oleuropein, cyclosporine A and ribavirin all showed no ability to inhibit SARS-CoV-2 CPE. Hydroxychloroquine, cyclohexamide, didemnin B, γ-mangostin and linoleic acid were all able to inhibit viral CPE at concentrations that did not induce cytotoxicity. However, these drugs inhibited CPE at concentrations that induced cytomorbidity, indicating non-specific anti-viral activity. Conclusions We describe the methodology for a simple in vitro drug screening assay that identifies potential anti-viral drugs via their ability to inhibit SARS-CoV-2-induced CPE. The additional growth assay illustrated how several drugs display anti-viral activity at concentrations that induce cytomorbidity. For instance, hydroxychloroquine showed anti-viral activity at concentrations that slow cell growth, arguing that its purported in vitro anti-viral activity arises from non-specific impairment of cellular activities. The cytomorbidity assay can therefore rapidly exclude potential false positives.

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