scholarly journals A Novel Gene, ROA, Is Required for Normal Morphogenesis and Discharge of Ascospores in Gibberella zeae

2010 ◽  
Vol 9 (10) ◽  
pp. 1495-1503 ◽  
Author(s):  
Kyunghun Min ◽  
Jungkwan Lee ◽  
Jin-Cheol Kim ◽  
Sang Gyu Kim ◽  
Young Ho Kim ◽  
...  
Keyword(s):  
Crop Residues ◽  
Turgor Pressure ◽  
Green Fluorescence Protein ◽  
Dispersal Distance ◽  
Gibberella Zeae ◽  
Multiple Roles ◽  
Head Blight ◽  
Content Type ◽  
Novel Gene ◽  
Fluorescence Protein

ABSTRACT Head blight, caused by Gibberella zeae, is a significant disease among cereal crops, including wheat, barley, and rice, due to contamination of grain with mycotoxins. G. zeae is spread by ascospores forcibly discharged from sexual fruiting bodies forming on crop residues. In this study, we characterized a novel gene, ROA, which is required for normal sexual development. Deletion of ROA (Δroa) resulted in an abnormal size and shape of asci and ascospores but did not affect vegetative growth. The Δroa mutation triggered round ascospores and insufficient cell division after spore delimitation. The asci of the Δroa strain discharged fewer ascospores from the perithecia but achieved a greater dispersal distance than those of the wild-type strain. Turgor pressure within the asci was calculated through the analysis of osmolytes in the epiplasmic fluid. Deletion of the ROA gene appeared to increase turgor pressure in the mutant asci. The higher turgor pressure of the Δroa mutant asci and the mutant spore shape contributed to the longer distance dispersal. When the Δroa mutant was outcrossed with a Δmat1-2 mutant, a strain that contains a green fluorescence protein (GFP) marker in place of the MAT1-2 gene, unusual phenotypic segregation occurred. The ratio of GFP to non-GFP segregation was 1:1; however, all eight spores had the same shape. Taken together, the results of this study suggest that ROA plays multiple roles in maintaining the proper morphology and discharge of ascospores in G. zeae.

scholarly journals Effects of Temperature and Moisture on Development of Fusarium graminearum Perithecia in Maize Stalk Residues

2015 ◽  
Vol 82 (1) ◽  
pp. 184-191 ◽  
Author(s):  
Valentina Manstretta ◽  
Vittorio Rossi
Keyword(s):  
Fusarium Graminearum ◽  
Crop Residues ◽  
Head Blight ◽  
Dispersal Patterns ◽  
Content Type ◽  
Predominant Component ◽  
Effects Of Temperature ◽  
Ascospore Production ◽  
Maize Stalks ◽  
Over Time

ABSTRACTFusarium graminearumis the predominant component of the Fusarium head blight complex of wheat.F. graminearumascospores, which initiate head infection, mature in perithecia on crop residues and become airborne. The effects of temperature (T) and moisture on perithecium production and maturation and on ascospore production on maize stalk residues were determined. In the laboratory, perithecia were produced at temperatures between 5 and 30°C (the optimum was 21.7°C) but matured only at 20 and 25°C. Perithecia were produced when relative humidity (RH) was ≥75% but matured only when RH was ≥85%; perithecium production and maturation increased with RH. Equations describing perithecium production and maturation over time as a function ofTand RH (R2> 0.96) were developed. Maize stalks were also placed outdoors on three substrates: a grass lawn exposed to rain; a constantly wet, spongelike foam exposed to rain; and a grass lawn protected from rain. No perithecia were produced on stalks protected from rain. Perithecium production and maturation were significantly higher on the constantly wet foam than on the intermittently wet lawn (both exposed to rain). Ascospore numbers but not their dispersal patterns were also affected by the substrate.

scholarly journals Novel Detection Strategy To Rapidly Evaluate the Efficacy of Antichlamydial Agents

2016 ◽  
Vol 61 (2) ◽  
Author(s):  
Yan Zhang ◽  
Yuqi Xian ◽  
Leiqiong Gao ◽  
Hiba Elaasar ◽  
Yao Wang ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Green Fluorescence Protein ◽  
Proof Of Concept ◽  
Reporter System ◽  
Content Type ◽  
Gfp Reporter ◽  
Fluorescence Protein ◽  
Novel Strategy ◽  
The Impact ◽  
Antimicrobial Effectiveness

ABSTRACT Chlamydia trachomatis infections present a major heath burden worldwide. The conventional method used to detect C. trachomatis is laborious. In the present study, a novel strategy was utilized to evaluate the impact of antimicrobial agents on the growth of C. trachomatis and its expression of ompA promoter-driven green fluorescence protein (GFP). We demonstrate that this GFP reporter system gives a robust fluorescent display of C. trachomatis growth in human cervical epithelial cells and, further, that GFP production directly correlates to changes in ompA expression following sufficient exposure to antimicrobials. Validation with azithromycin, the first-line macrolide drug used for the treatment of C. trachomatis infection, highlights the advantages of this method over the traditional method because of its simplicity and versatility. The results indicate both that ompA is highly responsive to antimicrobials targeting the transcription and translation of C. trachomatis and that there is a correlation between changing GFP levels and C. trachomatis growth. This proof-of-concept study also reveals that the ompA-GFP system can be easily adapted to rapidly assess antimicrobial effectiveness in a high-throughput format.

Comparing Genetic Variation within Red Winter Wheat Populations with and without Image-Based Optical Sorter Selection

2019 ◽  
Vol 9 (2) ◽  
pp. 266-277
Author(s):  
Hussein M. Khaeim ◽  
Anthony Clark ◽  
Tom Pearson ◽  
Dr. David Van Sanford
Keyword(s):  
Genetic Variation ◽  
Fusarium Head Blight ◽  
Heading Date ◽  
Fusarium Head Blight Resistance ◽  
Gibberella Zeae ◽  
Mass Selection ◽  
Head Blight ◽  
Red Winter Wheat ◽  
Two Populations ◽  
Head Scab

Head scab is historically a devastating disease affecting not just all classes of wheat but also barley and other small grains around the world. Fusarium head blight (FHB), or head scab, is caused most often by Fusarium graminearum (Schwabe), (sexual stage – Gibberella zeae) although several Fusarium spp. can cause the disease. This study was conducted to determine the effect of mass selection for FHB resistance using an image-based optical sorter. lines were derived from the C0 and C2 of two populations to compare genetic variation within populations with and without sorter selection. Our overall hypothesis is that sorting grain results in improved Fusarium head blight resistance. Both of the used wheat derived line populations have genetic variation, and population 1 has more than population 17. They are significantly different from each other for fusarium damged kernel (FDK), deoxynivalenol (DON), and other FHB traits. Although both populations are suitable to be grown for bulks, population 1 seems better since it has more genetic variation as well as lower FDK and DON, and earlier heading date. Lines within each population were significantly different and some lines in each population had significantly lower FDK and DON after selection using an optical sorter. Some lines had significant reduction in both FDK and DON, and some others had either FDK or DON reduction. Lines of population 1 that had significant reduction, were more numerous than in population 17, and FDK and DON reduction were greater.

scholarly journals Expansion of EasyClone-MarkerFree toolkit for Saccharomyces cerevisiae genome with new integration sites

2021 ◽  
Author(s):  
Mahsa Babaei ◽  
Luisa Sartori ◽  
Alexey Karpukhin ◽  
Dmitrii Abashkin ◽  
Elena Matrosova ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Recombinant Dna ◽  
Green Fluorescence Protein ◽  
Cellular Growth ◽  
Green Fluorescence ◽  
Biotechnological Production ◽  
Yeast Saccharomyces Cerevisiae ◽  
Integration Sites ◽  
Fluorescence Protein ◽  
Integration Efficiency

Abstract Biotechnological production requires genetically stable recombinant strains. To ensure genomic stability, recombinant DNA is commonly integrated into the genome of the host strain. Multiple genetic tools have been developed for genomic integration into baker's yeast Saccharomyces cerevisiae. Previously, we had developed a vector toolkit EasyClone-MarkerFree for stable integration into eleven sites on chromosomes X, XI, and XII of S. cerevisiae. The markerless integration was enabled by CRISPR-Cas9 system. In this study, we have expanded the kit with eight additional intergenic integration sites located on different chromosomes. The integration efficiency into the new sites was above 80%. The expression level of green fluorescence protein (gfp) for all eight sites was similar or above XI-2 site from the original EasyClone-MarkerFree toolkit. The cellular growth was not affected by the integration into any of the new eight locations. The eight-vector expansion kit is available from AddGene.

Establishment of an Embryotoxicity Assay with Green Fluorescence Protein-expressing Embryonic Cell-derived Cardiomyocytes

1999 ◽  
Vol 27 (3) ◽  
pp. 471-484 ◽  
Author(s):  
Susanne Bremer ◽  
Maaike Van Dooren ◽  
Martin Paparella ◽  
Eugen Kossolov ◽  
Bernd Fleischmann ◽  
...  
Keyword(s):  
Green Fluorescence Protein ◽  
Embryonic Cell ◽  
Green Fluorescence ◽  
Fluorescence Protein

scholarly journals Fertility of mice receiving vitrified adult mouse ovaries

Reproduction ◽  
2006 ◽  
Vol 131 (4) ◽  
pp. 681-687 ◽  
Author(s):  
Toshio Hani ◽  
Takanori Tachibe ◽  
Saburo Shingai ◽  
Nobuo Kamada ◽  
Otoya Ueda ◽  
...  
Keyword(s):  
Germ Cells ◽  
Adult Mouse ◽  
Green Fluorescence Protein ◽  
Green Fluorescence ◽  
Experimental Animals ◽  
Immature Mouse ◽  
Estrous Cyclicity ◽  
Fluorescence Protein ◽  
High Degree ◽  
Old Mice

Cryopreservation of the ovaries is a useful technology for preservation of germ cells from experimental animals, because if the female founder is infertile or has mutated mitochondrial DNA, preservation of female germ cells is necessary. Although it is possible to cryopreserve immature mouse ovaries with a high degree of viability by vitrification with a mixture of several cryoprotectants, the viability of cryopreserved adult mouse ovaries is still unknown. Here, we investigated the viability of mouse ovaries at various ages after cryopreservation by vitrification techniques. Donor ovaries were collected from 10-day-, 4-week-, 10-week- and 7-month-old, female, nulliparous, green fluorescence protein (GFP)-transgenic mice and cryopreserved by vitrification. The vitrified-warmed ovaries were orthotopically transplanted to 4- or 10-week-old mice. GFP-positive pups were obtained in all experimental groups. In the 4-week-old recipients, the percentages of GFP-positive pups among the total pups from recipients transplanted with ovaries of 10-day-, 4-week-, 10-week- and 7-month-old donors were 44%, 9%, 12% and 4% respectively. In the 10-week-old recipients, the percentages of GFP-positive pups among the total pups from recipients transplanted with ovaries of 10-day-, 4-week-, 10-week- and 7-month-old donors were 36%, 16%, 2% and 9% respectively. Furthermore, GFP-positive pups also were obtained from recipients transplanted with ovaries of donors without normal estrous cyclicity. Our results indicate that cryopreservation of mouse ovaries by vitrification is a useful method for the preservation of female germ cells from mice of various ages.

scholarly journals Survival and Inoculum Production of Gibberella zeae in Wheat Residue

Plant Disease ◽  
2004 ◽  
Vol 88 (7) ◽  
pp. 724-730 ◽  
Author(s):  
S. A. Pereyra ◽  
R. Dill-Macky ◽  
A. L. Sims
Keyword(s):  
Management Strategies ◽  
Soil Surface ◽  
Triticum Aestivum L ◽  
Gibberella Zeae ◽  
Effective Management ◽  
Head Blight ◽  
Fusarium Spp ◽  
Residue Decomposition ◽  
Inoculum Production ◽  
Mesh Bags

Survival and inoculum production of Gibberella zeae (Schwein.) Petch (anamorph Fusarium graminearum (Schwabe)), the causal agent of Fusarium head blight of wheat and barley, was related to the rate of wheat (Triticum aestivum L.) residue decomposition. Infested wheat residue, comprising intact nodes, internodes, and leaf sheaths, was placed in fiberglass mesh bags on the soil surface and at 7.5- to 10-cm and 15- to 20-cm depths in chisel-plowed plots and 15 to 20 cm deep in moldboard-plowed plots in October 1997. Residue was sampled monthly from April through November during 1998 and every 2 months through April to October 1999. Buried residue decomposed faster than residue placed on the soil surface. Less than 2% of the dry-matter residue remained in buried treatments after 24 months in the field, while 25% of the residue remained in the soil-surface treatment. Survival of G. zeae on node tissues was inversely related to the residue decomposition rate. Surface residue provided a substrate for G. zeae for a longer period of time than buried residue. Twenty-four months after the initiation of the trial, the level of colonization of nodes in buried residue was half the level of colonization of residue on the soil surface. Colonization of node tissues by G. zeae decreased over time, but increased for other Fusarium spp. Ascospores of G. zeae were still produced on residue pieces after 23 months, and these spores were capable of inducing disease. Data from this research may assist in developing effective management strategies for residues infested with G. zeae.

scholarly journals Depletion of Undecaprenyl Pyrophosphate Phosphatases Disrupts Cell Envelope Biogenesis in Bacillus subtilis

2016 ◽  
Vol 198 (21) ◽  
pp. 2925-2935 ◽  
Author(s):  
Heng Zhao ◽  
Yingjie Sun ◽  
Jason M. Peters ◽  
Carol A. Gross ◽  
Ethan C. Garner ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Transcriptional Repression ◽  
Cell Envelope ◽  
Turgor Pressure ◽  
Teichoic Acid ◽  
Lethal Gene ◽  
Genetic Redundancy ◽  
Wall Teichoic Acid ◽  
Content Type ◽  
Lipid Ii

ABSTRACTThe integrity of the bacterial cell envelope is essential to sustain life by countering the high turgor pressure of the cell and providing a barrier against chemical insults. InBacillus subtilis, synthesis of both peptidoglycan and wall teichoic acids requires a common C55lipid carrier, undecaprenyl-pyrophosphate (UPP), to ferry precursors across the cytoplasmic membrane. The synthesis and recycling of UPP requires a phosphatase to generate the monophosphate form Und-P, which is the substrate for peptidoglycan and wall teichoic acid synthases. Using an optimizedclusteredregularlyinterspacedshortpalindromicrepeat (CRISPR) system with catalytically inactive (“dead”)CRISPR-associated protein9(dCas9)-based transcriptional repression system (CRISPR interference [CRISPRi]), we demonstrate thatB. subtilisrequires either of two UPP phosphatases, UppP or BcrC, for viability. We show that a third predicted lipid phosphatase (YodM), with homology to diacylglycerol pyrophosphatases, can also support growth when overexpressed. Depletion of UPP phosphatase activity leads to morphological defects consistent with a failure of cell envelope synthesis and strongly activates the σM-dependent cell envelope stress response, includingbcrC, which encodes one of the two UPP phosphatases. These results highlight the utility of an optimized CRISPRi system for the investigation of synthetic lethal gene pairs, clarify the nature of theB. subtilisUPP-Pase enzymes, and provide further evidence linking the σMregulon to cell envelope homeostasis pathways.IMPORTANCEThe emergence of antibiotic resistance among bacterial pathogens is of critical concern and motivates efforts to develop new therapeutics and increase the utility of those already in use. The lipid II cycle is one of the most frequently targeted processes for antibiotics and has been intensively studied. Despite these efforts, some steps have remained poorly defined, partly due to genetic redundancy. CRISPRi provides a powerful tool to investigate the functions of essential genes and sets of genes. Here, we used an optimized CRISPRi system to demonstrate functional redundancy of two UPP phosphatases that are required for the conversion of the initially synthesized UPP lipid carrier to Und-P, the substrate for the synthesis of the initial lipid-linked precursors in peptidoglycan and wall teichoic acid synthesis.

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