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1.
J Exp Bot ; 59(9): 2371-8, 2008.
Article in English | MEDLINE | ID: mdl-18467324

ABSTRACT

Fusarium head blight (FHB; scab), primarily caused by Fusarium graminearum, is a devastating disease of wheat worldwide. FHB causes yield reductions and contamination of grains with trichothecene mycotoxins such as deoxynivalenol (DON). The genetic variation in existing wheat germplasm pools for FHB resistance is low and may not provide sufficient resistance to develop cultivars through traditional breeding approaches. Thus, genetic engineering provides an additional approach to enhance FHB resistance. The objectives of this study were to develop transgenic wheat expressing a barley class II chitinase and to test the transgenic lines against F. graminearum infection under greenhouse and field conditions. A barley class II chitinase gene was introduced into the spring wheat cultivar, Bobwhite, by biolistic bombardment. Seven transgenic lines were identified that expressed the chitinase transgene and exhibited enhanced Type II resistance in the greenhouse evaluations. These seven transgenic lines were tested under field conditions for percentage FHB severity, percentage visually scabby kernels (VSK), and DON accumulation. Two lines (C8 and C17) that exhibited high chitinase protein levels also showed reduced FHB severity and VSK compared to Bobwhite. One of the lines (C8) also exhibited reduced DON concentration compared with Bobwhite. These results showed that transgenic wheat expressing a barley class II chitinase exhibited enhanced resistance against F. graminearum in greenhouse and field conditions.


Subject(s)
Chitinases/metabolism , Fusarium/physiology , Genetic Engineering , Immunity, Innate , Plant Diseases/microbiology , Triticum/immunology , Triticum/microbiology , Chitinases/genetics , Gene Expression , Hordeum/enzymology , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/immunology , Plants, Genetically Modified/microbiology , Triticum/genetics
2.
Plant Cell Rep ; 27(7): 1217-25, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18392625

ABSTRACT

The number of viable shoots influences the overall architecture and productivity of wheat (Triticum aestivum L.). The development of lateral branches, or tillers, largely determines the resultant canopy. Tillers develop from the outgrowth of axillary buds, which form in leaf axils at the crown of the plant. Tiller number can be reduced if axillary buds are not formed or if the outgrowth of these buds is restricted. The teosinte branched1 (tb1) gene in maize, and homologs in rice and Arabidopsis, genetically regulate vegetative branching. In maize, increased expression of the tb1 gene restricts the outgrowth of axillary buds into lateral branches. In this study, the maize tb1 gene was introduced through transformation into the wheat cultivar "Bobwhite" to determine the effect of tb1 overexpression on wheat shoot architecture. Examination of multiple generations of plants reveals that tb1 overexpression in wheat results in reduced tiller and spike number. In addition, the number of spikelets on the spike and leaf number were significantly greater in tb1-expressing plants, and the height of these plants was also reduced. These data reveal that the function of the tb1 gene and genetic regulation of lateral branching via the tb1 mode of action is conserved between wheat, rice, maize and Arabidopsis. Thus, the tb1 gene can be used to alter plant architecture in agriculturally important crops like wheat.


Subject(s)
Genes, Plant/genetics , Plants, Genetically Modified/growth & development , Triticum/growth & development , Zea mays/genetics , Blotting, Southern , Flowers/genetics , Flowers/growth & development , Gene Expression Regulation, Plant , Meristem/genetics , Meristem/growth & development , Phenotype , Plant Leaves/genetics , Plant Leaves/growth & development , Plant Shoots/genetics , Plant Shoots/growth & development , Plants, Genetically Modified/genetics , Reverse Transcriptase Polymerase Chain Reaction , Triticum/genetics
3.
Plant Cell Rep ; 26(4): 479-88, 2007 Apr.
Article in English | MEDLINE | ID: mdl-17103001

ABSTRACT

Fusarium head blight (FHB) of wheat, caused by Fusarium graminearum and other Fusarium species, is a major disease problem for wheat production worldwide. To combat this problem, large-scale breeding efforts have been established. Although progress has been made through standard breeding approaches, the level of resistance attained is insufficient to withstand epidemic conditions. Genetic engineering provides an alternative approach to enhance the level of resistance. Many defense response genes are induced in wheat during F. graminearum infection and may play a role in reducing FHB. The objectives of this study were (1) to develop transgenic wheat overexpressing the defense response genes alpha-1-purothionin, thaumatin-like protein 1 (tlp-1), and beta-1,3-glucanase; and (2) to test the resultant transgenic wheat lines against F. graminearum infection under greenhouse and field conditions. Using the wheat cultivar Bobwhite, we developed one, two, and four lines carrying the alpha-1-purothionin, tlp-1, and beta-1,3-glucanase transgenes, respectively, that had statistically significant reductions in FHB severity in greenhouse evaluations. We tested these seven transgenic lines under field conditions for percent FHB disease severity, deoxynivalenol (DON) mycotoxin accumulation, and percent visually scabby kernels (VSK). Six of the seven lines differed from the nontransgenic parental Bobwhite line for at least one of the disease traits. A beta-1,3-glucanase transgenic line had enhanced resistance, showing lower FHB severity, DON concentration, and percent VSK compared to Bobwhite. Taken together, the results showed that overexpression of defense response genes in wheat could enhance the FHB resistance in both greenhouse and field conditions.


Subject(s)
Fusarium/growth & development , Plant Diseases/genetics , Triticum/genetics , Blotting, Southern , Blotting, Western , Glucan 1,3-beta-Glucosidase/genetics , Glucan 1,3-beta-Glucosidase/metabolism , Plant Diseases/microbiology , Plants, Genetically Modified , Reverse Transcriptase Polymerase Chain Reaction , Transformation, Genetic , Triticum/metabolism , Triticum/microbiology
4.
Plant Cell Rep ; 25(4): 313-9, 2006 Apr.
Article in English | MEDLINE | ID: mdl-16252090

ABSTRACT

Fusarium head blight (FHB), caused primarily by Fusarium graminearum, is a major disease problem in wheat (Triticum aestivum). Genetic engineering holds significant potential to enhance FHB resistance in wheat. Due to the requirement of screening for FHB resistance on flowers at anthesis, the number of screens carried out in a year is limited. Our objective was to evaluate the feasibility of using the rapid-maturing dwarf wheat cultivar Apogee as an alternative genotype for transgenic FHB resistance research. Our transformation efficiency (number of transgenic plants/number of embryos) for Apogee was 1.33%. Apogee was also found to exhibit high FHB susceptibility and reached anthesis within 4 weeks. Interestingly, microsatellite marker haplotype analysis of the chromosome 3BS FHB resistant quantitative trait locus (QTL) region indicated that this region maybe deleted in Apogee. Our results indicate that Apogee is particularly well suited for accelerating transgenic FHB resistance research and transgenic wheat research in general.


Subject(s)
Fusarium/physiology , Plant Diseases/genetics , Plant Diseases/microbiology , Triticum/genetics , Triticum/microbiology , Genetic Engineering , Models, Biological , Transformation, Genetic , Triticum/growth & development
5.
Mycopathologia ; 156(4): 329-32, 2003.
Article in English | MEDLINE | ID: mdl-14682459

ABSTRACT

Several inhibitors of deoxyhypusine synthase were examined for antifungal activity. All compounds, used at concentrations of 1.0 mM or less, reduced mycelial growth of the oat leaf stripe pathogen Pyrenophora avenae grown on solid media. Four of the compounds completely inhibited fungal growth, 4a doing so at a concentration of only 0.61 mM. Inhibition of fungal growth afforded by some of the compounds was more marked than was reflected by the radial growth measurements.


Subject(s)
Ascomycota/drug effects , Enzyme Inhibitors/pharmacology , Oxidoreductases Acting on CH-NH Group Donors/antagonists & inhibitors , Antifungal Agents/pharmacology , Ascomycota/enzymology
6.
Biochem J ; 373(Pt 3): 885-92, 2003 Aug 01.
Article in English | MEDLINE | ID: mdl-12737625

ABSTRACT

The role of spermine in the sensitivity of cells to various established and experimental anti-tumour agents was examined, using paired cell lines that possess or lack spermine synthase. All spermine-synthase-deficient cells had no detectable spermine, and elevated spermidine, content. Spermine content did not alter the cell growth rate. There was little or no difference in sensitivity of immortalized mouse embryonic fibroblasts to doxorubicin, etoposide, cisplatin, methylglyoxal bis(guanylhydrazone) or H(2)O(2) and only a slight increase in sensitivity to vinblastine and nocodazole. However, the absence of spermine clearly increased the sensitivity to 1,3-bis(2-chloroethyl)- N -nitrosourea, suggesting that depletion of spermine may be a useful way to increase the anti-neoplastic effects of anti-tumour agents that form chloroethyl-mediated interstrand DNA cross-links. The effects of spermine on the response to polyamine analogues (which have been proposed to be useful anti-neoplastic agents) were complex, and depended on the compound examined and on the cells tested. Sensitivity to CHENSpm ( N (1)-ethyl- N (11)-[(cycloheptyl)methyl]-4,8-diazaundecane) was substantially greater in immortalized fibroblasts that lack spermine. In contrast, BE-3-4-3 [ N (1), N (12)-bis(ethyl)spermine] and BE-3-3-3 [ N (1), N (11)-bis(ethyl)norspermine] were more active against cells that contained spermine. The presence of spermine correlated with a greater induction of spermidine/spermine- N (1)-acetyltransferase by BE-3-3-3, which is consistent with suggestions that this induction is important for the response to this drug. These findings support the concepts that different polyamine analogues have different sites of action and that CHENSpm has a different site of action from BE-3-3-3.


Subject(s)
Antineoplastic Agents/pharmacology , Spermine Synthase/metabolism , Animals , Base Sequence , Blotting, Western , Chromatography, High Pressure Liquid , DNA Primers , Male , Mice , Spermine Synthase/genetics , Transfection , Tumor Cells, Cultured
7.
FEBS Lett ; 527(1-3): 223-28, 2002 Sep 11.
Article in English | MEDLINE | ID: mdl-12220664

ABSTRACT

Activation of the extracellular signal-regulated kinases (ERKs) 1 and 2 is correlated to cell survival, but in some cases ERKs can act in signal transduction pathways leading to apoptosis. Treatment of mouse fibroblasts with 20 microM etoposide elicited a sustained phosphorylation of ERK 1/2, that increased until 24 h from the treatment in parallel with caspase activity. The inhibitor of ERK activation PD98059 abolished caspase activation, but caspase inhibition did not reduce ERK 1/2 phosphorylation, suggesting that ERK activation is placed upstream of caspases. Both ERK and caspase activation were blocked in cells depleted of polyamines by the ornithine decarboxylase inhibitor alpha-difluoromethylornithine (DFMO). In etoposide-treated cells, DFMO also abolished phosphorylation of c-Jun NH(2)-terminal kinases triggered by the drug. Polyamine replenishment with exogenous putrescine restored the ability of the cells to undergo caspase activation and ERK 1/2 phosphorylation in response to etoposide. Ornithine decarboxylase activity decreased after etoposide, indicating that DFMO exerts its effect by depleting cellular polyamines before induction of apoptosis. These results reveal a role for polyamines in the transduction of the death signal triggered by etoposide.


Subject(s)
Caspases/metabolism , Etoposide/pharmacology , Fibroblasts/metabolism , Mitogen-Activated Protein Kinases/metabolism , Polyamines/metabolism , Animals , Apoptosis/drug effects , Apoptosis/physiology , Caspase Inhibitors , Caspases/drug effects , Coumarins/pharmacology , Eflornithine/pharmacology , Enzyme Activation/drug effects , Enzyme Inhibitors/pharmacology , Fibroblasts/drug effects , Fibroblasts/pathology , Mice , Mitogen-Activated Protein Kinase 1/drug effects , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3 , Mitogen-Activated Protein Kinases/drug effects , Oligopeptides/pharmacology , Ornithine Decarboxylase Inhibitors , Phosphorylation , Putrescine/pharmacology , Signal Transduction
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