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Results for “stomatal phenotyping”

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At least 19 recordsLinked to original sources
PubMed2026

Virus-induced gene editing of stomatal regulators in Nicotiana benthamiana enables rapid functional genomics.

Virus-induced gene editing (VIGE) holds promise as a rapid and scalable approach for functional genomics in plants. Here, we apply a tobacco rattle virus (TRV)-based single-guide RNA (sgRNA) delivery system to target key regulators of stomatal development in Nicotiana benthamiana using transgenic Cas9-expressing lines. sgRNAs fused to a mobile RNA element and co-delivered with TRV enabled both somatic and heritable genome editing across orthologs of STOMAGEN, EPF2, YODA, and SPEECHLESS. Somatic editing frequencies reached up to 95%, and heritable tetra-allelic mutations were recovered in multiple target genes. Mutants exhibited significant, gene-specific changes in stomatal density, with corresponding effects on leaf temperature indicative of altered evaporative cooling. Additionally, sgRNAs fused to an AmCyan reporter enabled visualization of virus-infected tissues, allowing stomatal phenotyping in edited M0 sectors. This TRV-based platform facilitates functional assessment of genes influencing stomatal patterning and offers a powerful tool for dissecting gene function in a developmentally and physiologically relevant context.

Nicotiana↗
PubMed1976

Phenotypic mixing of vesicular stomatitis virus and D-type oncornavirus.

After mixed infection of cells with vesicular stomatitis virus (VSV, thermolabile mutant tl 17) and oncornavirus type D, phenotypically mixed virions are formed containing the VSV genome and oncornavirus and VSV envelope. The virions are thermostable and have serologic characteristics of both viruses.

Animals↗
PubMed1976

Biological and electron microscopic studies on the phenotypic mixing of the thermolabile mutant of vesicular stomatitis virus, tl-17, with avian RNA tumor viruses.

A thermolabile as well as thermosensitive mutant of vesicular stomatitis virus, VSV-tl-17, could be thermostabilized by phenotypic mixing with avian RNA tumor viruses (ATV). Biological, immunological and morphological studies revealed that this effect is due to a replacement of the thermolabile projections of the VSV by the avian tumor viral projections. The arrangement of projections of VSV and ATV on phenotypically mixed viria was studied by electron microscopy. A-type particles were detected in chick embryo cells which were doubly infected by ATV and VSV. Their intracytoplasmic appearance seemed to be the result of a disturbed maturation of ATV due to the relative deficiency of envelope proteins which were depleted by VSV maturation in the course of phenotypic mixing.

Alpharetrovirus↗
PubMed1979

Neutralization of pseudotypes of vesicular stomatitis virus by sera from avian retrovirus-infected hosts.

We investigated the capacity of lymphocytes and sera from chickens bearing tumors induced by avian sarcoma viruses (ASV) to interact with phenotypically mixed particles of vesicular stomatitis virus (VSV) and ASV. Immune chicken sera were able to specifically neutralize such VSV pseudotypes. This ability could be absorbed out, however, on purified preparations of avian retroviruses, suggesting that reactivity was primarily against avian retrovirus enveloped components. Supernatant fluids containing phenotypically mixed particles were unable to stimulate division of lymphocytes of tumor-bearing hosts, an ability possessed by culture fluids containing native ASV particles. Polyacrylamide gel analysis was unable to resolve any distinct pseudotype protein, which was not present in either of the parental virus types. Treatment with crude preparations of ultraviolet (UV)-irradiated VSV pseudotype material did not afford immunity against subsequent challenge with live ASV.

Alpharetrovirus↗
PubMed1979

A vesicular stomatitis virus (cytomegalovirus) pseudotype and its use in neutralization tests.

Infection with vesicular stomatitis virus (VSV) of human diploid cells preinfected with the AD-169 strain of human cytomegalovirus (CMV) resulted in the formation of a VSV (CMV) pseudotype. Its formation was favored by increasing the bicarbonate content in doubly-infected cultures. The pseudotype was capable of infecting not only human but also rabbit cells. Pseudotype particles formed after infection with the tl 17 mutant of VSV, which carries a thermolabile lesion in its neutralization antigen, were more stable at 45 degrees than the original tl 17 virus. The pseudotype was used in the neutralization test with human sera. All sera positive for CMV antibody in the complement-fixation (CF) test were also reactive in the neutralization test. In addition, numerous sera negative for CMV antibody in the CF test neutralized the pseudotype.

Adult↗
PubMed1977

Mechanism of restriction of ecotropic and xenotropic murine leukemia viruses and formation of pseudotypes between the two viruses.

Ecotropic and xenotropic murine leukemia viruses (MuLV's) constitute separate interference groups; within each group there is cross-interference, but between the groups there is no detectable interference. Interference is manifest against pseudotypes in which the vesicular stomatitis virus genome is contained within the coat of one of the murine leukemia viruses. The pseudotypes display the cell specificity of the leukemia viruses: pseudotypes with an ecotropic MuLV coat infect mouse cells but not rabbit or mink cells; pseudotypes with a xenotropic MuLV coat infect rabbit or mink cells well but mouse cells very poorly. Efficient pseudotype formation also occurs between the two MuLV classes, and both the interference patterns and the cell specificity of these pseudotypes are entirely determined by their envelope. Using these pseudotypes, ecotropic MuLV infection could be established in xenogeneic cells, and the resulting progeny could be scored by using a conventional XC cell assay. Also, xenotropic MuLV infection could be established in a mouse cell, showing that no absolute intracellular barrier against xenotropic virus growth exists in murine cells. The major barriers against both xenotropic and ecotropic MuLV therefore are cell surface barriers. Xenogeneic cells probably lack receptors for ecotropic MuLV, but murine cells may either lack receptors for xenotropic MuLV or have receptors that are blocked by endogenous expression of the glycoprotein of endogenous xenotropic MuLV.

Cell Line↗
PubMed2025

Smarter stomata: emergent technologies unlocking yield potential in a changing climate.

Stomata, the gatekeepers of leaf gas exchange, regulate carbon dioxide uptake and water loss, functions increasingly critical as crops face more frequent, intense heat and drought. Under dry conditions, stomatal conductance (g s) typically decreases, limiting carbon assimilation and yield. Heat stress, in contrast, elicits variable g S responses: sometimes increasing to facilitate transpirational cooling, while at other times decreasing, especially when combined with drought. Heat and drought also induce complex, context-dependent shifts in stomatal anatomy. Smaller, denser stomata improve drought resilience in some cases, while reduced density confers greater tolerance in others. The optimal stomatal ideotype remains unknown, and different or even opposing traits may confer resilience dependent on the environmental scenario. Substantial genotypic variation in g s and stomatal anatomy, high heritability and co-localized quantitative trait loci for stomatal traits and yield highlight their untapped potential as breeding targets for climate-resilient crops. However, stomatal traits remain largely absent from breeding pipelines due to challenges of phenotyping at scale. This is changing rapidly. Advances in deep learning, porometry, digital microscopy, and remote sensing now enable high-throughput measurement of stomatal physiology and anatomy. Next-generation breeding technologies including clustered regularly interspaced short palindromic repeats (CRISPR), multi-omics approaches, and artificial intelligence-driven ideotype selection models could revolutionize breeding, allowing precise engineering of stomatal traits for resilience to environmental stress. The time has come to move beyond characterizing stomatal traits and start actively incorporating them into breeding strategies. By leveraging these technologies, stomatal traits can become high value targets, unlocking their potential to enhance crop performance in a hotter, drier future.

abiotic stress↗
PubMed1978

Pseudotype particles of vesicular stomatitis virus with surface antigens of bovine leukaemia virus--VSV (BLV) -- as a sensitive probe for detecting antibodies in the sera of spontaneously infected cattle.

Phenotypically mixed particles containing the genome of vesicular stomatitis virus (VSV) and envelope antigen corresponding to bovine leukaemia virus (BLV) -- the VSV (BLV) pseudotypes -- can be employed as a rapid, specific and sensitive probe for detecting BLV-neutralizing antibodies in bovine sera.

Animals↗
PubMed1978

Coinfection with a rhabdovirus: vesicular stomatitis virus of Indiana and New-Jersey serotypes.

Coinfection of cells with vesicular stomatitis virus (VSV) of Indiana and New-Jersey serotypes were performed. Thermosensitive mutants (ts) of VSV Indiana and the wild type strain (+) of New-Jersey were used. Harvests and titrations were made at permissive(PT) and nonpermissive (NPT) temperatures. It was shown that the harvest was mainly composed of one parental-like infectious particles. The dominance of one serotype over the other was shown to be a function of the relative multiplicity of the two viruses; the presence of a thermosensitive lesion imparts a disadvantage to the corresponding serotype. Non parental-like particles were also detected. As expected, these particles were detected only in two conditions. 1) Harvest performed at NPT and titrations allowed at PT.- Most of the infectious particles (i.e. twin particles) resistant to anti-Nj serum developped a plaque (i.e. mixed-plaque)containing virions of both serotypes: Indiana (ts) and New-Jersey (+). After sonication or EDTA treatment of the harvest, prior to titrations, no more mixed-plaques were formed. Examination of the harvest by electron microscopy showed that 7-17 % of the particles formed aggregates; therefore, it is likely that the twin-particles are in fact aggregates. 2) Harvest performed at PT and titrations allowed at NPT.-It has been shown that 1 % of the wild type infectious particles was resistant to anti-Nj serum even though being of Nj genotype. It was inactivated by a mixture of anti-Nj and anti-In sera and therfore behave as pseudotypes. But since twin particles, when plated at Nt, would give rise to an homogenous progeny from New-Jersey (+), they could be confused with pseudotypes. Under those conditions there is no absolute evidence that phenotypic mixing really occurs between VSV of Indiana and New-Jersey serotypes.

Cell Line↗
PubMed1975

Further evidence for the existence of a viral envelope protein defect in the Bryan high-titer strain of Rous sarcoma virus.

Electron microscopy observations of purified Bryan high-titer Rous sarcoma virus (BH RSV) using the freeze-drying technique showed that progeny made in the absence of a helper virus lacked visible surface projections or spikes. Phenotypic mixing experiments employing BH RSV and a thermolabile mutant of vesicular stomatitis virus, tl 17, yielded no evidence of pseudotype formation. Since tl 17 is known to be defective for an envelope glycoprotein, the lack of successful phenotypic mixing with BH RSV is consistent with the observed absence of viral spikes.

Animals↗
PubMed1979

A rapid neutralization test for antibodies to bovine leukemia virus, with the use of rhabdovirus pseudotypes.

Two rhabdoviruses, vesicular stomatitis (type Indiana) and Chandipura viruses, formed pseudotype particles with envelope antigens provided by bovine leukemia virus (BLV). The pseudotypes are infectious for calf, human, mink, and rat cells, but the most sensitive indicator proved to be the Vero cells. Infectivity of the pseudotypes was increased by DEAE-dextran present during adsorption. Sera of spontaneously infected cattle contained high titers (some over 1/10,000) of antibodies neutralizing the pseudotypes, whereas sera of cattle from uninfected herds possessed no neutralizing activity in 1/10 dilution. The neutralization of these pseudotypes can serve as a rapid and sensitive test for the detection of antibodies in the cattle infected with BLV.

Antibodies, Viral↗