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Transformation: a tool for studying fungal pathogens of plants 总被引:18,自引:0,他引:18
Plant diseases caused by plant pathogenic fungi continuously threaten the sustainability of global crop production. An effective
way to study the disease-causing mechanisms of these organisms is to disrupt their genes, in both a targeted and random manner,
so as to isolate mutants exhibiting altered virulence. Although a number of techniques have been employed for such an analysis,
those based on transformation are by far the most commonly used. In filamentous fungi, the introduction of DNA by transformation
typically results in either the heterologous (illegitimate) integration or the homologous integration of the transforming
DNA into the target genome. Homologous integration permits a targeted gene disruption by replacing the wild-type allele on
the genome with a mutant allele on transforming DNA. This process has been widely used to determine the role of newly isolated
fungal genes in pathogenicity. The heterologous integration of transforming DNA causes a random process of gene disruption
(insertional mutagenesis) and has led to the isolation of many fungal mutants defective in pathogenicity. A big advantage
of insertional mutagenesis over the more traditional chemical or radiation mutagenesis procedures is that the mutated gene
is tagged by transforming DNA and can subsequently be cloned using the transforming DNA. The application of various transformation-based
techniques for fungal gene manipulation and how they have increased our understanding and appreciation of some of the most
serious plant pathogenic fungi are discussed.
Received 9 May 2001; received after revision 2 July 2001; accepted 3 July 2001 相似文献
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Horvat A Nikezić G Petrović S Kanazir DT 《Cellular and molecular life sciences : CMLS》2001,58(4):636-644
The subsynaptosomal distribution and specific binding of 17beta-estradiol in vitro to mitochondria isolated from presynaptic nerve endings of female rat brain were examined. 17Beta-estradiol is (i) distributed unequally in synaptosomes and mitochondria posses the highest capacity to bind estradiol with respect to the available amount of the hormone. (ii) Estradiol binds specifically to isolated synaptosomal mitochondria. A Michaelis-Menten plot of specific binding was sigmoidal within a concentration range of 0.1-5 nM of added estradiol, with a saturation plateau at 3 nM. Binding of higher estradiol concentrations demonstrated an exponential Michaelis-Menten plot, indicating non-specific binding to mitochondria. Vmax and Km for the sigmoidal-shape range were estimated as 46 +/- 6 fmol of estradiol/mg of mitochondrial proteins and 0.46 +/- 0.07 nM free estradiol respectively. (iii) Estradiol binding is not affected by the removal of ovaries. The results show that inhibition of Na-dependent Ca2+ efflux from mitochondria by estradiol occurs according to an affinity change of the translocator for Na+, at the same estradiol concentrations that show specific binding to mitochondrial membranes. These data imply that physiological concentrations of estradiol, acting on mitochondrial membrane properties, extragenomically modulate the mitochondrial, and consequently the synaptosomal content of Ca2+, and in that way exert a significant change in nerve cell homeostasis. 相似文献