4Prasher (42) cloned a cDNA for the green fluorescent protein (GFP) gene from the jellyfish Aequorea victoria in 1992. Shortly thereafter, to the amazement of many investigators, this gene or derivatives thereof were successfully expressed and conferred fluorescence to bacteria and Caenorhabditis elegans cells in culture (10, 31), followed by yeast (24,39), mammals (40), Drosophila (66), Dictyostelium (23,30), plants (28,49), and filamentous fungi (54). The tremendous success of GFP as a reporter can be attributed to unique qualities of this 238-amino-acid, 27-kDa protein which absorbs light at maxima of 395 and 475 nm and emits light at a maximum of 508 nm. The fluorescence of GFP requires only UV or blue light and oxygen, and therefore, unlike the case with other reporters (-glucuronidase, -galacturonidase, chloramphenicol acetyltransferase, and firefly luciferase) that rely on cofactors or substrates for activity, in vivo observation of gfp expression is possible with individual cells, with cell populations, or in whole organisms interacting with symbionts or environments in real time. Complications caused by destructive sampling, cell permeablization for substrates, or leakage of products do not occur. Furthermore, the GFP protein is extremely stable in vivo and has been fused to the C or N terminus of many cellular and extracellular proteins without a loss of activity, thereby permitting the tagging of proteins for gene regulation analysis, protein localization, or specific organelle labeling. The mature protein resists many proteases and is stable up to 65°C and at pH 5 to 11, in 1% sodium dodecyl sulfate or 6 M guanidinium chloride (reviewed in references 17 and 67), and in tissue fixed with formaldehyde, methanol, or glutaraldehyde. However, GFP loses fluorescence in methanol-acetic acid (3:1) and can be masked by autofluorescent aldehyde groups in tissue fixed with glutaraldehyde. Fluorescence is optimal at pH 7.2 to 8.0 (67).Limitations on GFP as a reporter for some applications are its low turnover rate, 2-h lag time for autoactivation of its chromophore, improper folding at high temperatures (37°C), which results in nonfluorescent and insoluble forms of the protein, and requirement for oxygen, which is not present in equal concentrations in all subcellular locations or cell types (reviewed in references 17 and 67). These characteristics of GFP, however, have not posed a problem for many applications, and mutant forms of GFP that have an ability to fold properly at high temperatures, increased solubility and fluorescence, reduced photobleaching (16, 17, 51), and reduced half-lives (1) have been developed. Coupled with fluorescenceactivated cell sorting, confocal microscopy or quantitative image analysis techniques, GFP technology can be used to isolate transformed cells or specific cell types from populations of cells (14), to quantify gene expression of individual cells within whole organisms (8), or to assess the dispersal and biomass of organisms in complex environments, such as in animal or plan...
We evaluated effects of both physical and biological components of the environment on growth of Pantoea agglomerans on inoculated pear and apple blossoms and on spread of the bacterium to blossoms on non-inoculated trees. The center three rows of 0.35- to 0.5-ha blocks of four pear cultivars and four apple cultivars were sprayed with a suspension of streptomycin-resistant P. agglomerans strain C9-1S (C9-1S) at 20 to 60% and 60 to 90% bloom. Cultivars were chosen to create a sequence of continuous bloom from late March (d'Anjou pear) through mid-May (Red Rome apple). Each cultivar block was quartered into plots; two plots were treated twice with streptomycin sulfate near mid- and full bloom to suppress populations of indigenous bacterial epiphytes and the other two plots were treated with water. Colonization of blossoms by C9-1S and by indigenous bacterial epiphytes were monitored on inoculated trees and along transects of noninoculated trees. Immediately after spraying, C9-1S was detected principally on blossoms sampled from inoculated trees. As bloom progressed, trees up to 18 m from inoculated trees had high proportions of blossoms colonized by C9-1S. Streptomycin significantly (P= 0.05) reduced incidence of isolation and size of detectable populations of culturable bacteria (indigenous bacteria plus C9-1S) from pear blossoms in 1998 and from apple blossoms in both 1998 and 1999, but the antibiotic treatment did not affect incidence of isolation, size of detectable populations, or spread of C9-1S compared to the water-treated control in any experiment. Across all cultivars, relative area under the curve for size of detectable populations of C9-1S on inoculated trees and for incidence of isolation of C9-1S from noninoculated trees was positively correlated with mean degree hours per day during bloom (r= 0.61 to 0.73) and negatively correlated with the proportion of days with rain (r = -0.79 to -0.84). The results indicate that establishment and growth of C9-1S on pome fruit flowers was not strongly affected by streptomycin or by competition from indigenous bacterial epiphytes and, as with Erwinia amylovora, temperature is an important environmental variable affecting successful spread of this biological control agent from blossom to blossom.
Eukaryotic viruses and bacteriophage have been implicated in disease and bleaching in corals, but the compositional and functional diversity of these viruses in healthy and compromised hosts remains underexplored. To investigate whether viral assemblages differ in concert with coral bleaching, we collected bleached and non-bleached conspecific pairs of corals during a minor bleaching event in 2016 from reefs on the island of Mo'orea, French Polynesia. Using electron microscopy (EM), we identified several viral particle types, all reminiscent of medium to large-sized nucleocytoplasmic large DNA viruses (NCLDV). We also found that viral metagenomes from bleached corals have significantly more eukaryotic virus sequences, whereas bacteriophage sequences are significantly more abundant in viral metagenomes from non-bleached colonies. In this study, we also initiated the assembly of the first eukaryotic dsDNA coral virus genome. Based on our EM imagery and our taxonomic annotations of viral metagenome sequences, we hypothesize that this genome represents a novel, phylogenetically distinct member of the NCLDVs, with its closest sequenced relative being a distant marine flagellate-associated virus. We also showed that this NCLDV is abundant in bleached corals, but it is also present in apparently healthy corals, suggesting it plays a role in the onset and/or severity of coral bleaching.
Xylella fastidiosa is a major threat to the worldwide agriculture industry (1, 2). Despite its global importance, many aspects of X. fastidiosa biology and pathogenicity are poorly understood.
In the face of new emerging respiratory viruses, such as SARS-CoV2, vaccines and drug therapies are not immediately available to curb the spread of infection. Non-pharmaceutical interventions, such as mask-wearing and social distance, can slow the transmission. However, both mask and social distance have not prevented the spread of respiratory viruses SARS-CoV2 within the US. There is an urgent need to develop an intervention that could reduce the spread of respiratory viruses. The key to preventing transmission is to eliminate the emission of SARS-CoV2 from an infected person and stop the virus from propagating in the human population. Rhamnolipids are environmentally friendly surfactants that are less toxic than the synthetic surfactants. In this study, rhamnolipid products, 222B, were investigated as disinfectants against enveloped viruses, such as bovine coronavirus and herpes simplex virus 1 (HSV-1). The 222B at 0.009% and 0.0045% completely inactivated 6 and 4 log PFU/mL of HSV-1 in 5–10 min, respectively. 222B at or below 0.005% is also biologically safe. Moreover, 50µl of 222B at 0.005% on ~1 cm2 mask fabrics or plastic surface can inactivate ~ 103 PFU HSV-1 in 3–5 min. These results suggest that 222B coated on masks or plastic surface can reduce the emission of SARS-CoV2 from an infected person and stop the spread of SARS-CoV2.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.