scholarly journals Yellow nutsedge WRI3/4-like gene improves drought tolerance in Arabidopsis thaliana by promoting cuticular wax biosynthesis

2019 ◽  
Author(s):  
Chao Cheng ◽  
Shutong Hu ◽  
Yun Han ◽  
Di Xia ◽  
Bang-Lian Huang ◽  
...  
Keyword(s):  
Fatty Acid Biosynthesis ◽  
Phylogenetic Analyses ◽  
Cuticular Wax ◽  
Wild Type ◽  
Yellow Nutsedge ◽  
Transgenic Lines ◽  
In The Wild ◽  
Peg Treatment ◽  
Simulated Drought ◽  
Tag Accumulation

Abstract In this study we cloned a WRI1-like gene from yellow nutsedge. Conserved domain and phylogenetic analyses indicated it to be a WRI3/4-like gene. Arabidopsis plants transformed with WRI3/4-like gene showed significantly improved tolerance to both PEG-simulated drought stress and real dehydration compared with the wild type. Quantitative RT-PCR indicated that, under unstressed conditions, the expressions of key genes involved in fatty acid biosynthesis was not significantly different between wild type (WT) and transgenic lines, while the expressions of genes involved in cuticular wax biosynthesis was significantly higher in transgenic lines compared with the wild type. The PEG treatment slightly decreased the expression of above mentioned genes in WT plants while it was significantly increased in transgenic lines compared with their respective unstressed control. Without PEG treatment, the expression of TAG1, the gene involved in triacylglycerol (TAG) accumulation, was 10-40% lower in the transgenic lines than that in the wild type. However, after PEG treatment, the expression of TAG1 was slightly decreased in the wild type, while in the transgenic lines its expression was decreased by 20-70% compared with unstressed transgenic lines and was highly significantly lower than that in the wild type. The cuticular wax content in Arabidopsis leaves was significantly higher in the transgenic lines than that in the wild type, while the oil content was not significantly different.

scholarly journals Yellow nutsedge WRI3/4-like gene improves drought tolerance in Arabidopsis thaliana by promoting cuticular wax biosynthesis

2019 ◽  
Author(s):  
Chao Cheng ◽  
Shutong Hu ◽  
Yun Han ◽  
Di Xia ◽  
Bang-Lian Huang ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Drought Stress ◽  
Drought Tolerance ◽  
Normal Growth ◽  
Growth Conditions ◽  
Cuticular Wax ◽  
Expression Data ◽  
Yellow Nutsedge ◽  
Transgenic Lines ◽  
Simulated Drought

Abstract Using RACE PCR, full length WRI1-like gene was amplified from yellow nutsedge. Conserved domain and phylogenetic analyses suggested it as WRI3/4-like gene. Tissue-specific expression data showed the highest expression in leaves, followed by roots while the lowest expression was detected in tuber. Transgenic Arabidopsis plants expressing nutsedge WRI3/4-like gene showed significantly improved tolerance to both PEG-simulated drought stress and real dehydration, compared with the wild type (WT). Under normal growth conditions, the expressions of key fatty acid biosynthesis genes was not significantly different between WT and transgenic lines, while the expressions of genes involved in cuticular wax biosynthesis was significantly higher in transgenic lines compared with the WT. The PEG-simulated drought stress did not induce any significant change in the expression of fatty acid and wax biosynthesis genes in WT plants, while the expression of fatty acid and wax biosynthesis genes was significantly increased in transgenic lines compared with WT as well as unstressed transgenic control. The expression of TAG1, the gene involved in triacylglycerol (TAG) accumulation, was significantly lower in the transgenic lines than that in the WT in normal growth conditions. Drought stress slightly decreased the expression of TAG1 in the WT, but significantly lowered it in transgenic lines compared with its unstressed transgenic control and WT. Consistent with gene expression data, the cuticular wax content in Arabidopsis leaves was significantly higher in the transgenic lines than in the WT, while the oil content was not significantly different. Our results indicated that WRI3/4-like gene from Cyperus esculentus improves drought tolerance in Arabidopsis probably by promoting cuticular wax biosynthesis and, hence, could be a valuable target for improving drought tolerance in crops through recombinant DNA technology.

scholarly journals Role of Unsaturated Fatty Acid Biosynthesis in Virulence of Streptococcus mutans

2007 ◽  
Vol 75 (3) ◽  
pp. 1537-1539 ◽  
Author(s):  
Elizabeth M. Fozo ◽  
Kathy Scott-Anne ◽  
Hyun Koo ◽  
Robert G. Quivey
Keyword(s):  
Fatty Acids ◽  
Streptococcus Mutans ◽  
Type Strain ◽  
Wild Type Strain ◽  
Fatty Acid Biosynthesis ◽  
Wild Type ◽  
Acid Biosynthesis ◽  
Carious Lesions ◽  
In The Wild

ABSTRACT An insertionally inactivated fabM strain of Streptococcus mutans does not produce unsaturated membrane fatty acids and is acid sensitive (E. M. Fozo and R. G. Quivey, Jr., J. Bacteriol. 186:4152-4158, 2004). In this study, the strain was shown to be poorly transmissible from host to host. Animals directly infected with the fabM strain exhibited fewer and less severe carious lesions than those observed in the wild-type strain.

scholarly journals Yellow nutsedge WRI4-like gene improves drought tolerance in Arabidopsis thaliana by promoting cuticular wax biosynthesis

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Chao Cheng ◽  
Shutong Hu ◽  
Yun Han ◽  
Di Xia ◽  
Bang-Lian Huang ◽  
...  
Keyword(s):  
Drought Stress ◽  
Drought Tolerance ◽  
Transgenic Arabidopsis ◽  
Cuticular Wax ◽  
Cyperus Esculentus ◽  
Expression Data ◽  
Yellow Nutsedge ◽  
Transgenic Lines ◽  
Use Efficiency ◽  
Key Genes

Abstract Background Cuticular wax plays important role in protecting plants from drought stress. In Arabidopsis WRI4 improves drought tolerance by regulating the biosynthesis of fatty acids and cuticular wax. Cyperus esculentus (yellow nutsedge) is a tough weed found in tropical and temperate zones as well as in cooler regions. In the current study, we report the molecular cloning of a WRI4-like gene from Cyperus esculentus and its functional characterization in Arabidopsis. Results Using RACE PCR, full-length WRI-like gene was amplified from yellow nutsedge. Phylogenetic analyses and amino acid comparison suggested it to be a WRI4-like gene. According to the tissue-specific expression data, the highest expression of WRI4-like gene was found in leaves, followed by roots and tuber. Transgenic Arabidopsis plants expressing nutsedge WRI4-like gene manifested improved drought stress tolerance. Transgenic lines showed significantly reduced stomatal conductance, transpiration rate, chlorophyll leaching, water loss and improved water use efficiency (WUE). In the absence of drought stress, expression of key genes for fatty acid biosynthesis was not significantly different between transgenic lines and WT while that of cuticular wax biosynthesis genes was significantly higher in transgenic lines than WT. The PEG-simulated drought stress significantly increased expression of key genes for fatty acid as well as wax biosynthesis in transgenic Arabidopsis lines but not in WT plants. Consistent with the gene expression data, cuticular wax load and deposition was significantly higher in stem and leaves of transgenic lines compared with WT under control as well as drought stress conditions. Conclusions WRI4-like gene from Cyperus esculentus improves drought tolerance in Arabidopsis probably by promoting cuticular wax biosynthesis and deposition. This in turn lowers chlorophyll leaching, stomatal conductance, transpiration rate, water loss and improves water use efficiency under drought stress conditions. Therefore, CeWRI4-like gene could be a good candidate for improving drought tolerance in crops.

scholarly journals Transgenic Ectopic Overexpression of Broad Complex (BrC-Z2) in the Silk Gland Inhibits the Expression of Silk Fibroin Genes of Bombyx mori

Insects ◽  
2020 ◽  
Vol 11 (6) ◽  
pp. 374
Author(s):  
Jiangshan Cong ◽  
Cuicui Tao ◽  
Xuan Zhang ◽  
Hui Zhang ◽  
Tingcai Cheng ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Developmental Stages ◽  
Expression Patterns ◽  
Silk Gland ◽  
Silk Protein ◽  
Regulation Mechanism ◽  
Wild Type ◽  
Transgenic Lines ◽  
In The Wild ◽  
Broad Complex

Bombyx mori silk protein genes are strictly turned on and off in different developmental stages under the hormone periodically change. The broad complex (BrC) is a transcription factor mediating 20-hydroxyecdysone action, which plays important roles during metamorphosis. Here, we observed that two isoforms of BmBrC (BmBrC-Z2 and BmBrC-Z4) exhibited contrasting expression patterns with fibroin genes (FibH, FibL and P25) in the posterior silk gland (PSG), suggesting that BmBrC may negatively regulate fibroin genes. Transgenic lines were constructed to ectopically overexpress BmBrC-Z2 in the PSG. The silk protein genes in the transgenic line were decreased to almost half of that in the wild type. The silk yield was decreased significantly. In addition, the expression levels of regulatory factors (BmKr-h1 and BmDimm) response to juvenile hormone (JH) signal were inhibited significantly. Then exogenous JH in the BmBrC-Z2 overexpressed lines can inhibit the expression of BmBrC-Z2 and activate the expression of silk protein genes and restore the silk yield to the level of the wild type. These results indicated that BmBrC may inhibit fibroin genes by repressing the JH signal pathway, which would assist in deciphering the comprehensive regulation mechanism of silk protein genes.

scholarly journals Aging-related variation of cuticular hydrocarbons in wild type and variant Drosophila melanogaster

2021 ◽  
Author(s):  
Jérôme Cortot ◽  
Jean-Pierre Farine ◽  
Jean-François Ferveur ◽  
Claude Everaerts
Keyword(s):  
Cuticular Hydrocarbons ◽  
Adult Life ◽  
Environmental Harm ◽  
Wild Type ◽  
Related Variation ◽  
Transgenic Lines ◽  
In The Wild ◽  
Large Period ◽  
Qualitative Pattern ◽  
Species Specific

Abstract The cuticle of all insects is covered with hydrocarbons which have multiple functions. Cuticular hydrocarbons (CHCs) basically serve to protect insects against environmental harm and to reduce dehydration. In many species, some CHCs also act as pheromones. CHCs have been intensively studied in Drosophila species and more specially in D. melanogaster. In this species, flies produce about 40 CHCs forming a complex sex- and species-specific bouquet. The quantitative and qualitative pattern of the CHC bouquet was characterized during the first days of adult life but remains unexplored in aging flies. Here, we characterized CHCs during the whole—or a large period of—adult life in males and females of several wild type and transgenic lines. Both types of lines included standard and variant CHC profiles. Some of the genotypes tested here showed very dramatic and unexpected aging-related variation based on their early days profile. This study provides a concrete dataset to better understand the mechanisms underlying the establishment and maintenance of CHCs on the fly cuticle. It could be useful to determine physiological parameters, including age and response to climate variation, in insects collected in the wild.

scholarly journals The Nicotiana tabacum Mediator subunit MED25 regulates nicotine biosynthesis through interacting with the basic helix-loop-helix (bHLH) transcription factor NtMYC2s

2021 ◽  
Author(s):  
Ge Bai ◽  
Yong Li ◽  
Da-Hai Yang ◽  
Tao Pang ◽  
Zhi-Yong Fan ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Acid Treatment ◽  
Plant Secondary Metabolites ◽  
Bhlh Transcription Factor ◽  
Wild Type ◽  
Helix Loop Helix ◽  
Transgenic Lines ◽  
In The Wild ◽  
Nicotine Biosynthesis

Nicotine is one of the most important secondary metabolites in tobacco, and its biosynthesis can be induced by topping and jasmonic acid treatment. NtMYC2s play pivotal roles in the regulation of nicotine. The mediator server as a bridge betwen the transcription factors and RNA polymerase in order to facilitates transcription and functions in plants. However, the role of mediator in the regulation of nicotine biosynthesis remains unknown. In this study, we firstly identify the NtMED25 through homologous analysis. NtMED25 interacts with NtMYC2s through the MD region. Interestingly, the nicotine content is decreased in the the knock-down transgenic lines of NtMED25, and the expression levels of two nicotine biosynthesis genes, NtQPT2 and NtPMT2, are also reduced when compared with that in the wild-type plants. Furthermore, NtMED25 enhances the binding of NtMYC2a/ NtMYC2b to the promoter of NtPMT2 and NtQPT2, and then facilitates the nicotine biosynthesis. Therefore, our study revealed the function of mediator in the regulation of nicotine, and provide the insight role on the transcriptional regulation of plant secondary metabolites.

scholarly journals Lytic Polysaccharide Monooxygenase from Talaromyces amestolkiae with an Enigmatic Linker-like Region: The Role of This Enzyme on Cellulose Saccharification

2021 ◽  
Vol 22 (24) ◽  
pp. 13611
Author(s):  
Juan Antonio Méndez-Líter ◽  
Iván Ayuso-Fernández ◽  
Florian Csarman ◽  
Laura Isabel de Eugenio ◽  
Noa Míguez ◽  
...  
Keyword(s):  
Catalytic Domain ◽  
Phylogenetic Analyses ◽  
Carbohydrate Binding ◽  
Wild Type ◽  
Wild Type Enzyme ◽  
Lytic Polysaccharide Monooxygenase ◽  
In The Wild ◽  
Type Sequence ◽  
Polysaccharide Monooxygenase

The first lytic polysaccharide monooxygenase (LPMO) detected in the genome of the widespread ascomycete Talaromyces amestolkiae (TamAA9A) has been successfully expressed in Pichia pastoris and characterized. Molecular modeling of TamAA9A showed a structure similar to those from other AA9 LPMOs. Although fungal LPMOs belonging to the genera Penicillium or Talaromyces have not been analyzed in terms of regioselectivity, phylogenetic analyses suggested C1/C4 oxidation which was confirmed by HPAEC. To ascertain the function of a C-terminal linker-like region present in the wild-type sequence of the LPMO, two variants of the wild-type enzyme, one without this sequence and one with an additional C-terminal carbohydrate binding domain (CBM), were designed. The three enzymes (native, without linker and chimeric variant with a CBM) were purified in two chromatographic steps and were thermostable and active in the presence of H2O2. The transition midpoint temperature of the wild-type LPMO (Tm = 67.7 °C) and its variant with only the catalytic domain (Tm = 67.6 °C) showed the highest thermostability, whereas the presence of a CBM reduced it (Tm = 57.8 °C) and indicates an adverse effect on the enzyme structure. Besides, the potential of the different T. amestolkiae LPMO variants for their application in the saccharification of cellulosic and lignocellulosic materials was corroborated.

scholarly journals The α-Gliadins in Bread Wheat: Effect of Nitrogen Treatment on the Expression of the Major Celiac Disease Immunogenic Complex in Two RNAi Low-Gliadin Lines

2021 ◽  
Vol 12 ◽  
Author(s):  
Susana Sánchez-León ◽  
María José Giménez ◽  
Francisco Barro
Keyword(s):  
Celiac Disease ◽  
Autoimmune Disorder ◽  
N Fertilization ◽  
Wild Type ◽  
Worldwide Population ◽  
Full Complement ◽  
Transgenic Lines ◽  
In The Wild ◽  
New Varieties ◽  
N Treatment

Celiac Disease (CD) is an autoimmune disorder that affects approximately 1% of the worldwide population. The α-gliadins of wheat contain the 33-mer peptide, the most active peptide in CD both in adults and pediatric patients. In this study, we have characterized the variants and expression profile of an α-gliadins amplicon, harboring the 33-mer peptide, in two low-gliadin RNAi wheat lines, under two different Nitrogen (N) treatments. We estimated that the amplicon expands 45 different α-gliadin variants with high variability due to length, randomly distributed SNPs, and the presence of encoded CD epitopes. Expression of this amplicon is reduced in both RNAi lines in comparison to the wild type. High N treatment significantly increases transcripts of the amplicon in the wild type, but not in the transgenic lines. Classification of α-gliadin variants, considering the number of epitopes, revealed that amplicon variants containing the full complement of 33-mer peptide were affected by N treatment, increasing their expression when N was increased. Line D793 provided higher and more stable silencing through different N fertilization regimes, expressing fewer CD epitopes than D783. Results of this study are important for better understanding of RNAi α-gliadin silencing in response to N treatments, and for undertaking new strategies by RNAi or CRISPR/Cas toward obtaining new varieties suitable for people suffering gluten intolerances.

Freeze-substituted fungal cells of Nectria haematococca: A comparison of the macroconidial walls of the adhesion-competent wild type with an adhesion-reduced mutant

1990 ◽  
Vol 48 (3) ◽  
pp. 688-689
Author(s):  
Thecan Caesar-Ton That ◽  
Lynn Epstein
Keyword(s):  
Freeze Substitution ◽  
Uranyl Acetate ◽  
Wild Type ◽  
Nectria Haematococca ◽  
Fruit Extract ◽  
Dialysis Tubing ◽  
Tube Emergence ◽  
Contrast Microscopy ◽  
In The Wild ◽  
Freeze Damage

Nectria haematococca mating population I (anamorph, Fusarium solani) macroconidia attach to its host (squash) and non-host surfaces prior to germ tube emergence. The macroconidia become adhesive after a brief period of protein synthesis. Recently, Hickman et al. (1989) isolated N. haematococca adhesion-reduced mutants. Using freeze substitution, we compared the development of the macroconidial wall in the wild type in comparison to one of the mutants, LEI.Macroconidia were harvested at 1C, washed by centrifugation, resuspended in a dilute zucchini fruit extract and incubated from 0 - 5 h. During the incubation period, wild type macroconidia attached to uncoated dialysis tubing. Mutant macroconidia did not attach and were collected on poly-L-lysine coated dialysis tubing just prior to freezing. Conidia on the tubing were frozen in liquid propane at 191 - 193C, substituted in acetone with 2% OsO4 and 0.05% uranyl acetate, washed with acetone, and flat-embedded in Epon-Araldite. Using phase contrast microscopy at 1000X, cells without freeze damage were selected, remounted, sectioned and post-stained sequentially with 1% Ba(MnO4)2 2% uranyl acetate and Reynold’s lead citrate. At least 30 cells/treatment were examined.

scholarly journals Rad51-Independent Interchromosomal Double-Strand Break Repair by Gene Conversion Requires Rad52 but Not Rad55, Rad57, or Dmc1

2007 ◽  
Vol 28 (3) ◽  
pp. 897-906 ◽  
Author(s):  
Thomas J. Pohl ◽  
Jac A. Nickoloff
Keyword(s):  
Gene Conversion ◽  
Strand Break ◽  
Double Strand Break ◽  
Single Strand ◽  
Homology Search ◽  
Wild Type ◽  
Dsb Repair ◽  
Single Strand Annealing ◽  
In The Wild ◽  
Break Induced Replication

ABSTRACT Homologous recombination (HR) is critical for DNA double-strand break (DSB) repair and genome stabilization. In yeast, HR is catalyzed by the Rad51 strand transferase and its “mediators,” including the Rad52 single-strand DNA-annealing protein, two Rad51 paralogs (Rad55 and Rad57), and Rad54. A Rad51 homolog, Dmc1, is important for meiotic HR. In wild-type cells, most DSB repair results in gene conversion, a conservative HR outcome. Because Rad51 plays a central role in the homology search and strand invasion steps, DSBs either are not repaired or are repaired by nonconservative single-strand annealing or break-induced replication mechanisms in rad51Δ mutants. Although DSB repair by gene conversion in the absence of Rad51 has been reported for ectopic HR events (e.g., inverted repeats or between plasmids), Rad51 has been thought to be essential for DSB repair by conservative interchromosomal (allelic) gene conversion. Here, we demonstrate that DSBs stimulate gene conversion between homologous chromosomes (allelic conversion) by >30-fold in a rad51Δ mutant. We show that Rad51-independent allelic conversion and break-induced replication occur independently of Rad55, Rad57, and Dmc1 but require Rad52. Unlike DSB-induced events, spontaneous allelic conversion was detected in both rad51Δ and rad52Δ mutants, but not in a rad51Δ rad52Δ double mutant. The frequencies of crossovers associated with DSB-induced gene conversion were similar in the wild type and the rad51Δ mutant, but discontinuous conversion tracts were fivefold more frequent and tract lengths were more widely distributed in the rad51Δ mutant, indicating that heteroduplex DNA has an altered structure, or is processed differently, in the absence of Rad51.

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