Effect of glyphosate on tryptamine production and Sekiguchi lesion formation in rice infected with Magnaporthe grisea

When exposed to light, the Sekiguchi lesion (sl) rice mutant has an enhanced resistance to Magnaporthe grisea infection responsible for Sekiguchi lesion formation and tryptamine accumulation. Glyphosate [N-(phosphonomethyl) glycine] pretreatment suppressed Sekiguchi lesion formation and tryptamine accumulation in the sl mutant after M. grisea infection even under light. This inhibition by glyphosate was blocked by the supply of exogenous tryptophan, but not by exogenous phenylalanine. In glyphosate-pretreated leaves, 5-enol-pyruvyl-shikimate-3-phosphate synthase gene expression and tryptophan biosynthesis were significantly suppressed. During tryptophan starvation, catalase activity was maintained at a high level even under light, leading to the suppression of H₂O₂ generation and DNA fragmentation. These results show a strong relationship between the tryptophan and tryptamine pathways in the induction of light-enhanced resistance to M. grisea infection in the sl mutant.     

Cytological responses in the hypersensitive reaction in cotyledon and stem tissues of Brassica napus after infection by Leptosphaeria maculans

A hypersensitive response (HR) occurs after infection of cotyledon, leaf and stem of Brassica napus cv. Surpass 400 by an avirulent strain of Leptosphaeria maculans. The cotyledon or the sixth true-leaf stages of plants were inoculated with pycnidiospores of L. maculans (strain UWA P11). For the first time in this specific pathosystem and its HR, we report condensation of cytoplasm, shrinkage in cell size and nuclear DNA fragmentation in cotyledon tissues, and in stem tissues, shrinkage and condensation of the cytoplasm, chromatin fragmentation and lobing of the nucleus.     

核盘菌自噬相关基因SsATG5和SsATG8的功能分析

为探究自噬在核盘菌Sclerotinia sclerotiorum致病过程中的作用,利用酵母Saccharomyces自噬相关基因（autophagy-related gene,ATG）编码的蛋白序列比对核盘菌基因组,获得核盘菌假定ATG,并以核盘菌1980菌株为出发菌株,基于同源重组的原理对假定ATG进行敲除和回补,并测定不同突变体的生长表型和致病能力。结果表明,从核盘菌基因组中比对到2个ATG,分别命名为SsATG5和SsATG8,两者在核盘菌致病过程中均上调表达。SsATG5和SsATG8敲除突变体在菌丝生长、产草酸和侵染垫形成方面与野生型菌株无明显差异,但SsATG5敲除突变体在离体拟南芥Arabidopsis thaliana叶片上的致病力显著下降了约40%,在活体拟南芥植株上的致病力显著下降了约80%,同时SsATG5回补突变体恢复了正常的致病力。表明SsATG5参与了核盘菌的致病过程,证实自噬在核盘菌致病过程中发挥着重要作用。     

Indole derivatives enhance resistance against the rice blast fungus <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

In leaves pretreated with an indole derivative [indole-3-acetic acid (IAA) tryptamine, or tryptophan], blast lesion formation was suppressed compared to those treated with distilled water (DW) as a control. Phenylalanine ammonia-lyase (PAL) activity and PAL expression were significantly enhanced in the IAA- or tryptophan-pretreated leaves, but not in tryptamine-pretreated leaves. This induction of resistance was inhibited by pretreatment with a PAL inhibitor, α-aminooxyacetic acid, in IAA- and in tryptophan-treated leaves, but not in tryptamine-treated leaves. This study strongly shows that the indole derivatives IAA tryptamine and tryptophan can enhance a disease resistance mechanism that is supported by different metabolic pathways.     

Virulence, accumulation of acetyl-coenzyme A, and pectate lyase synthesis are controlled by PhoP-PhoQ two-component regulatory system responding to organic acids of Erwinia chrysanthemi 3937

The PhoP-PhoQ two-components regulatory system is involved in the pathogenesis of animal, plant, and insect pathogenic bacteria in response to various environmental factors. To elucidate how this system contributes to the plant pathogenesis of Erwinia chrysanthemi 3937 (Ech 3937), marker-exchanged mutants of phoP and phoQ were constructed. Their role in the regulation of a major virulent factor, pectate lyase (Pel), in response to various organic acids was then tested. These mutants synthesized more Pel than did the wild type in the medium containing acetate or citrate as the sole source of carbon, but they synthesized less Pel than did the wild type in pyruvate or malate as the sole source of carbon. Synthesis of Pel did not differ in succinate, fumarate, or glycerol from the wild type. The phoP and phoQ mutants grown and resuspended in acetate or citrate also caused more maceration, and the wild type pretreated in pyruvate or malate caused more maceration than did the mutants. The level of intracellular acetyl-coenzyme A (acetyl-CoA) almost paralleled the synthesis of Pel in the wild type and in the mutants of the phoP and phoQ. These results suggested that acetyl-CoA may be involved in regulation of Pel synthesis through two-independent regulatory cascades via the PhoP-PhoQ system (in an opposite manner) in response to acetate/citrate and pyruvate/malate. However, ackA and pta genes, involved in the synthesis of acetyl-CoA in Escherichia coli, were not expressed as predicted on the basis of the level of acetyl-CoA. Thus there may be an additional regulation or pathway for the synthesis of acetyl-CoA in Ech 3937.     

Flagella-mediated motility is required for biofilm formation by Erwinia carotovora subsp. carotovora

To elucidate the role of flagella in biofilm formation by Erwinia carotovora subsp. carotovora EC1, we used a nonflagellate, nonmotile mutant (ΔfliC) and a flagellate, nonmotile mutant (ΔmotA). A biofilm-inducing medium, which contains the yeast peptone (YP) medium plus the salts of M-63 minimal medium, supported biofilm formation to a greater extent than either the YP or Luria Bertani (LB) medium alone. We demonstrated that both the ΔfliC and ΔmotA mutants greatly reduced their ability to form a biofilm on the surface of the wells of polyvinyl chloride (PVC) microtiter plates. The inability of both mutants to form biofilm on the PVC surface was further confirmed with phase-contrast microscopy. Both aflagellate (ΔfliC) and flagellate (ΔmotA) nonmotile mutants were equally defective in attachment to the PVC surface. The treatment of bacteria with the protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP), which inhibits the motility of this organism, reduced greatly the biofilm formation. Based on these results, flagella-mediated motility may play an important role in biofilm formation of E. carotovora subsp. carotovora EC1.     

Genetic diversity of the entomopathogen<Emphasis Type="Italic">Verticillium lecanii</Emphasis> on the basis of vegetative compatibility

Forty-three isolates ofVerticillium lecanii from insects, phytopathogenic fungi and other substrates were tested for vegetative compatibility by observing heterokaryon formation among complementary nitrate-nonutilizing (nit) mutants.nit mutants were isolated from 42/43 strains examined. Twenty-one isolates were self-incompatible, and the remaining 21 isolates were divided into 14 vegetative compatibility groups (VCGs): ten containing only a single strain each, and the remaining four containing two to four isolates each. Members of isolates in each of these VCGs all shared the same IGS haplotype. Further, the isolates within a VCG were correlated with one another in part by fragment patterns of mt-LrDNA, -SrDNA, Bt-2 and H4 region, by PCR-RFLP and -SSCP, but not by dsRNA. Two isolates belonging to VL-J2 have high virulence to aphids, whereas strains from VL-J1 lack this character. These findings indicate that two VCGs (VL-J1 and -J2) may originate from two distinct clonal lineages. Alternatively, high VCG diversity and HSI frequency ofV. lecanii might be associated with an array of distinct lineages. These data not only suggest relationships among DNA polymorphisms, virulence, and VCG, but also demonstrate genetic heterogeneity ofV. lecanii. http://www.phytoparasitica.org posting Sept. 30, 2003.     

Studies on red light-induced resistance of broad bean to Botrytis cinerea: I. Possible production of suppressor and elicitor by germinating spores of pathogen

When detached broad bean leaves were preinoculated with virulent strain B304 of Botrytis cinerea 24 h before a challenge inoculation with strain B304, lesion formation by B304 was significantly inhibited in red light but not in the dark. In leaves that were preinoculated with avirulent strain 021 and then challenged by B304, however, lesion formation was not inhibited even under red light. Such differences in lesion formation after the challenge inoculation with an avirulent strain were also observed with lesions caused by Alternaria alternata, a nonpathogen of broad bean and by avirulent strain 021R in the presence of germination fluid from spores of strains B304 and 021R. These results suggest the possibility that virulent B. cinerea produced a suppressor involved in induced susceptibility and an elicitor involved in resistance induced by red light during spore germination.     

Effects of some indole-related compounds on the infection behavior of Magnaporthe grisea

Based on their effect on the infection behavior of Magnaporthe grisea, indole-related compounds were classified into three groups. The first group, including tryptophan, indole-3-butyric acid, indole-3-pyruvic acid, and indole-3-acetamide, did not inhibit infection behavior such as spore germination, appressorium formation, or infection hypha formation in M. grisea. The second, including indole acetic acid, indole-3-acetonitrile, oxindole, and tryptamine inhibited all stages of infection behaviors in a dose-dependent manner. The third, including gramine and indole, did not inhibit spore germination or appressorium formation, whereas it did inhibit infection hypha formation in a dose-dependent manner. These results suggest that endogenous or exogenously applied indole-related compounds in the second and third groups may contribute to protection in blast-susceptible plants such as rice and barley.     

Effect of Phenylpyrrole-resistance Mutations on Ecological Fitness of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and their Genetical Basis in <Emphasis Type="Italic">Ustilago maydis</Emphasis>

Mutants of Botrytis cinerea and Ustilago maydis highly resistant to fludioxonil were isolated at a high frequency, after nitrosoguanidine or UV mutagenesis, respectively, and selection on media containing fludioxonil. Tests on the response of mutant strains to high osmotic pressure resulted in the identification of two fludioxonil-resistant phenotypes (FLD_osm/s and FLD_osm/r), regarding the sensitivity to high osmolarity. Approximately 95% of fludioxonil-resistant mutants were found to be more sensitive to high osmotic pressure than the wild-type parent strains. Genetic analysis of phenylpyrrole-resistance in the phytopathogenic basidiomycete U. maydis, showed that fludioxonil-resistance was coded by three unlinked chromosomal loci (U/fld-1, U/fld-2 and U/fld-3), from which only the U/fld-1 mutation coded an osmotic sensitivity similar to that of the wild-types. Cross-resistance studies with fungicides from other chemical groups showed that the mutations for resistance to phenylpyrroles affect the sensitivity of mutant strains to the aromatic hydrocarbon and dicarboximide fungicides, but not to the benzimidazoles, anilinopyrimidines, phenylpyridinamines, hydroxyanilides or the sterol biosynthesis inhibiting fungicides. A study of fitness parameters in the wild-type and fludioxonil-resistant mutants of B. cinerea, showed that all osmotic sensitive (B/FLD_osm/s) isolates had significant reductions in the characteristics determining saprophytic fitness such as mycelial growth, sporulation, conidial germination and sclerotial production. Contrary to that, with the exception of mycelial growth, the fitness parameters were unaffected or only slightly affected in most of the osmotic resistant (B/FLD_osm/r) isolates. Tests on cucumber seedlings showed that the osmotic-sensitive strains were significantly less pathogenic compared with the wild-type and B/FLD_osm/r strains of B. cinerea. Preventative applications of the commercial products Saphire 50 WP (fludioxonil) and Rovral 50 WP (iprodione) were effective against lesion development on cotyledons by the wild-type and the mutant strains of B. cinerea that were resistant to the anilinopyrimidine cyprodinil (B/CPL-27) and to the hydroxyanilide fenhexamid (B/FNH-21), but ineffective, even at high concentrations, against disease caused by the fludioxonil-resistant isolates (B/FLD) and a mutant strain resistant to the dicarboximide iprodione (B/IPR-1). Experiments on the stability of the fludioxonil-resistant phenotype showed a reduction of resistance, mainly in osmotic-sensitive isolates, when the mutants were grown on inhibitor-free medium. A rapid recovery of the high resistance was observed after mutants were returned to the selection medium. Studies on the competitive ability of mutant isolates against the wild-type parent strain of B. cinerea, by applications of a mixed conidial population, showed that, in vitro, all mutants were less competitive than the wild-type strain. However, the competitive ability of osmotic-resistant mutants was higher than the osmotic-sensitive ones. Furthermore, competition tests, in planta, showed a significant reduction of the frequency of both phenylpyrrole-resistant phenotypes, with a respective increase in the population of the wild-type strain of the pathogen.     

Flagellin from Pseudomonas syringae pv. tabaci induced hrp-independent HR in tomato

We have previously shown that flagellin of Pseudomonas syringae pv. tabaci is an elicitor that induces a hypersensitive reaction (HR) in nonhost tomato cells. Flagellin is the major HR elicitor produced by this pathogen, as shown by the inability of a flagellin-defective mutant, ΔfliC, to induce HR. Also, a ΔfliD mutant that secretes large amounts of monomer flagellins induces a strong HR in tomato. In this study, the possible involvement of an Hrp type III secretion system (TTSS) in flagellin-induced HR was investigated using flagella-defective mutants or Hrp TTSS-defective mutants. The hrcC gene encodes HrcC protein, which is required for Hrp pilus formation in the outer membrane. An hrcC mutation, introduced into the wild-type, ΔfliC, and ΔfliD mutants of P. syringae pv. tabaci did not affect swimming motility or flagellin secretion, whereas all ΔhrcC, ΔfliC, and ΔfliD mutants lost the ability to cause disease on host tobacco leaves. However, the ΔhrcC mutant and the ΔfliD/ΔhrcC double mutant were still able to induce HR cell death, expression of one of the defense-related genes hsr203J, and the generation of hydrogen peroxide in nonhost tomato cells. Thus, flagellin is required for both pathogenicity in host tobacco and HR in nonhost tomato. On the other hand, hrp TTSS is necessary for pathogenicity on host tobacco but is not indispensable to induce HR in nonhost tomato. These results clearly show that flagellin-induced HR is hrp-independent in tomato.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB049570     

Characterization of Laboratory Mutants of Botrytis cinerea Resistant to QoI Fungicides

Mutants of Botrytis cinerea with moderate and high resistance to pyraclostrobin, a Qo inhibitor of mitochondrial electron transport at the cytochrome bc ₁ complex, were isolated at a high mutation frequency, after nitrosoguanidine mutagenesis and selection on medium containing pyraclostrobin and salicylhydroxamate (SHAM), a specific inhibitor of cyanide-resistant (alternative) respiration. Oxygen uptake in whole cells was strongly inhibited in the wild-type strain by pyraclostrobin and SHAM, but not in the mutant isolates. Cross-resistance studies with other Qo and Qi inhibitors (QoIs and QiIs) of cytochrome bc ₁ complex of mitochondrial respiration showed that the mutation(s) for resistance to pyraclostrobin also reduced the sensitivity of mutant strains to other QoIs as azoxystrobin, fluoxastrobin, trifloxystrobin and picoxystrobin, but not to famoxadone and to the QiIs cyazofamid and antimycin-A. An increased sensitivity of pyraclostrobin-resistant strains to the carboxamide boscalid, an inhibitor of complex II, and to the anilinopyrimidine cyprodinil, a methionine biosynthesis inhibitor, was observed. Moreover, no effect of pyraclostrobin resistance mutation(s) on fungitoxicity of the hydroxyanilide fenhexamid, the phenylpyrrole fludioxonil, the benzimidazole benomyl, and to the phenylpyridinamine fluazinam, which affect other cellular pathways, was observed. Study of fitness parameters in the wild-type and pyraclostrobin-resistant mutants of B. cinerea showed that most mutants had a significant reduction in the sporulation, conidial germination and sclerotia production. Experiments on the stability of the pyraclostrobin-resistant phenotype showed a reduction of resistance, mainly in moderate resistant strains, when the mutants were grown on inhibitor-free medium. However, a rapid recovery of the resistance level was observed after the mutants were returned to a selective medium. Studies on the competitive ability of mutant isolates against the wild-type parent strain, by applications of a mixed conidial population, showed that, in vitro, all mutants were less competitive than the wild-type strain. However, the competitive ability of high resistant mutants was higher than the moderate ones. Pathogenicity tests on cucumber seedlings showed that all mutant strains tested exhibited an infection ability similar with the wild-type parent strain. Preventive applications of the commercial product of F-500 25EC (pyraclostrobin) were effective against lesion development on cotyledons by the wild-type, but ineffective, even at high concentrations, against disease caused by the pyraclostrobin-resistant isolates. Boscalid (F-510 50WG) was found equally effective against the disease caused by the wild-type or pyraclostrobin-resistant mutants. This is the first report indicating the appearance of B. cinerea strains resistant to QoI fungicides by the biochemical mechanism of site modification and the risk for field resistance.     

Vegetative compatibility groups inVerticillium dahliae isolates from cotton in Turkey

Verticillium dahliae Kleb. with a complicated genetic diversity is a widely distributed major pathogen resulting in cotton wilt, which causes high economic losses in cotton lint production in the cotton belt of Turkey. A collection of 70 TurkishV. dahliae isolates (68 from wilted cotton plants in 28 districts and two from watermelon plants in two districts) were tested for vegetative compatibility by observing heterokaryon formation among complementary nitrate-nonutilizing (nit) mutants. The mutants were tested against international reference tester isolates and also were paired with one another. Thirty-nine isolates were assigned to vegetative compatibility group (VCG) 2B, 19 to VCG2A and three to VCG4B. One isolate was self-incompatible and eight others could not be assigned to any of the identified VCGs because theirnit mutants showed negative reactions with the tester isolates of four VCGs or theirnit mutants reverted back to the wild type. This is the first report of VCGs inV. dahliae from cotton in Turkey.     

Note: Evaluation of antibacterial activity of essential oil of<Emphasis Type="Italic">Rosa damascena</Emphasis> on<Emphasis Type="Italic">Erwinia amylovora</Emphasis>

Significant antibacterial activity was observed in the essential oil (E.O.) ofRosa damascena Mill. and the Minimum Bactericidal Concentration (MBC) of the E.O. was determined as 1386.5 μg ml⁻¹ forErwinia amylovora, the causal agent of fire blight disease. Ooze formation on immature pears and lesion formation in artificially inoculated shoots were completely (100%) prevented by the essential oil ofR. damascena (1500 μg ml⁻¹), essential oil ofThymbra spicata var.spicata (500 μg ml⁻¹) and streptomycin (100 μg ml⁻¹). Copper oxychloride plus maneb significantly reduced ooze formation and lesion formation, but the control was less than that obtained with the essential oils or streptomycin. The essential oil ofR. damascena may be a useful natural bactericide for the control of the fire blight pathogen,E. amylovora. http://www.phytoparasitica.org posting July 14, 2004.     

苹果树腐烂病菌染色质重塑因子Vmles4的功能研究

染色质重塑因子INO80是一类由多亚基构成的遗传学调控因子，调控多种DNA代谢，在基因的表达中发挥着重要的调控功能。但其在苹果树腐烂病菌（Valsa mali）中是否存在及其生物学功能并不清楚，为此，本研究从病菌基因组中分析鉴定到INO80的一个亚基基因Vmles4，利用qRT-PCR技术分析了Vmles4在病菌侵染过程中的表达模式，利用Double-joint PCR技术和PEG介导的原生质体转化方法进行了基因敲除，然后对3个突变体的营养生长及致病力进行测定和分析，并对病菌的非核糖体肽合成酶基因VmNRPS12及VmNRPS14在突变体中的表达进行定量分析。结果表明：Vmles4在侵染初期的表达显著上调，接种后6 h上调表达6.2倍。敲除突变体的生长速率平均降低28%且菌丝生长稀疏，在富士苹果品种（Malus domestic cv. Fuji）叶片和枝条上的致病力分别降低到22.5%和27.5%，VmNRPS12及VmNRPS14基因在Vmles4突变体侵染苹果枝条24 h后的表达量分别下调86.5%和50%；综上所述，Vmles4正调控腐烂病菌的营养生长、致病力以及次级代谢合成酶基因VmNRPS12和VmNRPS14的表达。     

Multiple copies of <Emphasis Type="Italic">AMT2</Emphasis> are prerequisite for the apple pathotype of <Emphasis Type="Italic">Alternaria alternata</Emphasis> to produce enough AM-toxin for expressing pathogenicity

The apple pathotype of Alternaria alternata produces the cyclic depsipeptide AM-toxin and causes Alternaria blotch of apple. Previously, we cloned AMT2 from the apple pathotype as an orthologue of AFTS1, which is required for biosynthesis of the decatrienoic acid ester AF-toxin I of the strawberry pathotype. These genes were predicted to encode aldo-keto reductases involved in biosynthesis of a common precursor, 2-hydroxy-isovaleric acid, of AF-toxin I and AM-toxin. In this study, we analyzed the function of AMT2 in AM-toxin biosynthesis in the apple pathotype. DNA gel blot analysis of the apple pathotype strain IFO8984 with five restriction enzymes suggested that this strain has a single copy of AMT2 in the genome. However, gene disruption experiments showed that IFO8984 probably has three copies of AMT2. We made mutants having one or two copies of AMT2 disrupted. The single-copy mutants produced less AM-toxin than did the wild type and were still as pathogenic as the wild type. The two-copy mutants produced trace or undetectable amounts of AM-toxin and were markedly reduced in pathogenicity. Thus, AMT2 was verified to be required for AM-toxin biosynthesis and hence pathogenicity. The fact that the two-copy mutants have a remaining copy of AMT2 suggests that multiple copies of AMT2 are prerequisite for the pathogen to produce enough AM-toxin for full pathogenicity.     

Erwinia amylovora hrpN mutants,blocked in harpin synthesis,express a reduced virulence on host plants and elicit variable hypersensitive reactions on tobacco

Erwinia amylovora is the bacterium responsible for fire blight, a necrotic disease affecting many rosaceous plants and especially pear tree and apple tree. A protein named harpin, secreted through the Hrp secretion pathway and able to elicit an hypersensitive reaction (HR) on tobacco has recently been isolated. Mutants inhrpN, the gene encoding harpin were described as non pathogenic on immature pear fruit and unable to elicit an HR on tobacco [Weiet al., 1992; Wei and Beer, 1993]. In this paper, the phenotype on plant ofhrpN mutants was carefully determined.hrpN mutants expressed a weak but significant virulence on host plants. Furthermore, when infiltrated into tobacco leaf mesophyll, thehrpN mutants elicited varied responses that fluctuated from null reaction to full necrosis of the infiltrated area. These results show that harpin is not absolutely required neither for pathogenicity on host plant nor for elicitation of an hypersensitive reaction on tobacco. Furthermore, in all the tests performed, mutant blocked in harpin secretion remained non pathogenic and unable to elicit an HR on tobacco. This suggests that factor(s), different from harpin, involved both in pathogenicity and HR eliciting ability is (are) secreted through the Hrp secretion pathway.Abbreviations HR hypersensitive reaction - NSI necrosis severity index - CFU colonie forming units     

Insertional mutagenesis of <Emphasis Type="Italic">Magnaporthe grisea</Emphasis> toward decreased responsiveness of alternative respiration to inhibition by azoxystrobin

 Inhibitors of respiration with high affinity to the Qo site of cytochrome b constitute a major class of modern agricultural fungicides. Many fungal organisms, including plant pathogens, can circumvent this inhibition site by expressing alternative respiration, a pathway dependent on alternative oxidase and other unidentified gene products. The restriction enzyme-mediated insertion (REMI) technique was employed in this study to disrupt genes involved in the expression of fully functional alternative respiration of Magnaporthe grisea. In one of the REMI mutants obtained, the rescue response mediated by alternative respiration was completely abolished. In two other mutants, the response was diminished but not entirely silenced. For all three mutants, phenotype changes were not explained by an altered structure of the alternative oxidase gene AOXMg or by a decreased level of gene expression in response to the Qo inhibitor azoxystrobin. The gene potentially affected in one of the REMI mutants was a homolog of the ABC1 family of genes encoding chaperone-like proteins with roles in the optimal assembly of mitochondrial membrane complexes. Received: May 28, 2002 / Accepted: October 16, 2002 Acknowledgments The research was funded, in part, by financial support to C. Avila-Adame provided by the Fulbright-Garcia Robles Foundation, the Institute of International Education, CONACYT-Mexico, and the Colegio de Postgraduados-Mexico.     

A Recessive Lesion Mimic Mutant of Rice with Elevated Resistance to Fungal Pathogens

A gamma-ray-induced rice mutant, M1009, which spontaneously forms brown specks on leaves was isolated. In appearance, these lesions very much resemble those that are produced during the hypersensitive resistance response of rice to pathogens. A similar phenotype has been associated with a number of mutants in maize or Arabidopsis, which have been briefly categorized as disease lesion mimics. Brown speck formation was suppressed at temperatures of 25°C and above, and was severe at 20°C. Light irradiation is also required to initiate brown specks. In addition, the mutant exhibits heightened resistance to two major fungal pathogens of rice, Magnaporthe grisea and Cochliobolus miyabeanus. Genetic characterization of the mutant has indicated that its les-bs (lesion-brown speck) phenotype segregates as a monogenic recessive mutation. Received 13 September 1999/ Accepted in revised form 15 December 1999