Avirulence effectors(Avrs),encoded by plant pathogens,can be recognized by plants harboring the corresponding resistance proteins,thereby initiating effector-triggered immunity(ETI).In susceptible plants,however,Avrs ...Avirulence effectors(Avrs),encoded by plant pathogens,can be recognized by plants harboring the corresponding resistance proteins,thereby initiating effector-triggered immunity(ETI).In susceptible plants,however,Avrs can function as effectors,facilitating infection via effector-triggered susceptibility(ETS).Mechanisms of Avr-mediated ETS remain largely unexplored.Here we report that the Magnaporthe oryzae effector Avr-PikD enters rice cells via the canonical cytoplasmic secretion pathway and suppresses rice basal defense.Avr-PikD interacts with an LSD1-like transcriptional activator AKIP30 of rice,and AKIP30 is also a positive regulator of rice immunity,whereas Avr-PikD impedes its nuclear localization and suppresses its transcriptional activity.In summary,M.oryzae delivers Avr-PikD into rice cells to facilitate ETS by inhibiting AKIP30-mediated transcriptional regulation of immune response against M.oryzae.展开更多
[Objectives]This study was conducted to screen out rice resources resistant to rice blast(Magnaporthe oryzae).[Methods]The qualitative and quantitative resistance of 1659 rice resources from 45 countries and regions t...[Objectives]This study was conducted to screen out rice resources resistant to rice blast(Magnaporthe oryzae).[Methods]The qualitative and quantitative resistance of 1659 rice resources from 45 countries and regions to rice blast were evaluated by disease nursery in upland condition and the test of the spectrum to rice blast isolates.[Results]There were 292 entries which accounted for 17.6%showed high blast resistance(0 and 1 disease scale),68 entries(counted for 4.1%)showed resistance to blast(3 disease scale);and the number of the entries showed intermediate resistance,intermediate susceptible and susceptible were 208(with the corresponding percentage of 12.5%),471(28.4%),620(37.4%)respectively.Among the tested entries,27 entries including BG1222,BL122,BTX,IR37704-131-2-3-2,and LEBONNET had showed broad-spectrum blast resistance with the resistance frequency of higher than 90%,Quantitative resistance evaluation was conducted on some key resources,and 14 entries,of which are BR27,DRAGO,IR100,QINLIUAI、SERIBU GANTANG,YUEXIANGZHAN and so on,showed good quantitative resistances,and 8 entries had higher quantitative resistances than IR36.[Conclusions]This study provides important blast resistance resources for the local rice breeding program and has a significant value for the discovery of new blast resistance genes and its application in the blast resistance breeding.展开更多
In this study, three wheat varieties were tested to determine seed germination and the incidence of Magnaporthe oryzae triticum (MoT). Among these varieties, BARI Gom 24 (Prodip) wheat seed exhibited the highest seed ...In this study, three wheat varieties were tested to determine seed germination and the incidence of Magnaporthe oryzae triticum (MoT). Among these varieties, BARI Gom 24 (Prodip) wheat seed exhibited the highest seed germination rate (93%) but also had the highest incidence (30%) of MoT. To manage blast disease in an ecofriendly manner, seven treatments were employed: T<sub>1</sub> = Control, T<sub>2</sub> = Garlic clove extracts, T<sub>3</sub> = Aloe vera leaf extracts, T<sub>4</sub> = Black cumin seed extracts, T<sub>5</sub> = Neem leaf extracts, T<sub>6</sub> = Nativo 75 WG, and T<sub>7</sub> = Provax 200 WP. The experiment was conducted using a Randomized Complete Block Design (RCBD) layout with three replications using Prodip wheat variety that exhibited highest MoT infection severity based on laboratory analysis among collected varieties. Data were collected on blast disease incidence (%), disease severity, and various growth and yield parameters of wheat. The experiment’s results indicated that among all the treatments, T<sub>7</sub> (Seed treatment with Provax 200 WP) and T<sub>5</sub> (Foliar spraying with Neem leaf extract) performed better in controlling blast disease in wheat. The lowest blast disease incidence (%) was observed with T<sub>7</sub> (Provax 200 WP), with values of 7.86, 9.86, and 10.19 recorded during the milking stage, soft dough stage, and hard dough stage of wheat, respectively. T<sub>5</sub> (Neem leaf extract) also demonstrated a statistically equivalent reduction in blast disease incidence (%). In terms of disease severity, T<sub>7</sub> (Seed treatment with Provax 200 WP) showed the lowest values of 1.03, 1.23, and 1.63 during the milking stage, soft dough stage, and hard dough stage of wheat, respectively. Foliar spraying with neem leaf extract also exhibited similar result as of Provax 200 WP regarding panicle blast severity. As a result of these findings, it can be concluded that T<sub>5</sub> (Neem leaf extract) is recommended as an ecofriendly management approach for blast disease in wheat.展开更多
A mature appressorium cDNA library of rice blast fungus, Magnaporthegrisea, was constructed in a λ TriplEx2 vector by SMARTTM cDNA library containing 2.37×10^6 independent clones about 100% of which harbor forei...A mature appressorium cDNA library of rice blast fungus, Magnaporthegrisea, was constructed in a λ TriplEx2 vector by SMARTTM cDNA library containing 2.37×10^6 independent clones about 100% of which harbor foreign cDNA inserts with average size of 660 bp. Of 9 randomly selected clones, 2 expressed sequence tags (ESTs) sequences did not have homologous EST sequences of M. grisea in Gen Bank. The appressorium cDNA library is suitable for gene expression analysis and function analysis of the late stages of appressorium formation and the early stages of penetration of M. grisea.展开更多
377 isolates of Magnaporthe grisea were collected from 17 provinces in China and their geographic distribution of mating types and their fertility was tested with four standard isolates, KA3 and TH12 (Mat1.1) and Guy1...377 isolates of Magnaporthe grisea were collected from 17 provinces in China and their geographic distribution of mating types and their fertility was tested with four standard isolates, KA3 and TH12 (Mat1.1) and Guy11 and TH16 (Mat1.2) provided by CIRAD. 73 fertile isolates were tested with SCAR markers of 13 pairs of primers. Preliminary results showed that the geographic distribution of M. grisea existed among isolates collected from the same location as well as different locations and the genetic relationship between fertile isolates of the fungus in China. The existence of sexual reproduction of M. grisea was explored in the field as well.展开更多
Wheat blast,caused by the fungus Magnaporthe oryzae Triticum(MoT)pathotype,is a devastating disease persistent in South America and Bangladesh.Since MoT generally fails to cause visual symptoms in wheat until the head...Wheat blast,caused by the fungus Magnaporthe oryzae Triticum(MoT)pathotype,is a devastating disease persistent in South America and Bangladesh.Since MoT generally fails to cause visual symptoms in wheat until the heading stage when the infection would have advanced,disease control by fungicide application solely based on the detection of visual symptoms is ineffective.To develop an accurate and sensitive method to detect MoT at the seedling and vegetative stages for disease control,we sequenced the genomes of two MoT isolates from Brazil and identified two DNA fragments,MoT-6098 and MoT-6099,that are present in the MoT genome but not in the genome of the rice-infecting Magnaporthe oryzae Oryzae(MoO)pathotype.Using polymerase chain reaction(PCR),we confirmed the specificity of the two markers in 53 MoT and MoO isolates from South America and Bangladesh.To test the efficiency of the two markers,we first established a loop-mediated isothermal amplification(LAMP)method to detect MoT at isothermal conditions,without the use of a PCR machine.Following this,we used the Cas12a protein and guide RNAs(gRNAs)to target the MoT-6098 and MoT-6099 sequences.The activated Cas12a showed indiscriminate single-stranded deoxyribonuclease(ssDNase)activity.We then combined targetdependent Cas12a ssDNase activation with recombinase polymerase amplification(RPA)and nucleic acid lateral flow immunoassay(NALFIA)to develop a method that accurately,sensitively,and cost-effectively detects MoT-specific DNA sequences in infected wheat plants.This novel technique can be easily adapted for the rapid detection of wheat blast and other important plant diseases in the field.展开更多
Race-specific resistance and field resistance of 30 rice blast resistance monogenic lines derived from different resources were evaluated. The spectra of resistance to 163 Magnaporthe grisea isolates collected from in...Race-specific resistance and field resistance of 30 rice blast resistance monogenic lines derived from different resources were evaluated. The spectra of resistance to 163 Magnaporthe grisea isolates collected from indica rice in Guangdong Province, China ranged from 0.6% to 89.6%. Most of the monogenic lines showed a narrow resistance spectrum and high susceptibility in rice blast area, whereas the lines with Pikh and Pi1(t) had the broad resistance spectra of 89.6% and 82.2% respectively, showing a high and stable blast resistance in fields. According to the cluster analysis of specific resistance to 163 blast isolates tested, the 30 monogenic lines were divided into 15 groups, and based on the principal factor analysis, nine kinds of race-specific resistance were identified. Pik, Piz5, Pi9 and Pish can be used as candidate resistance genes for rice breeding since their specific resistance differed from those of the backbone parents in Guangdong, China. Gene pyramiding of Pikh [or Pi1(t)], Pi9 (or Piz5) and Pish (or Pita2) will be effective to obtain broad-spectrum blast resistance in rice breeding program in Guangdong, China. The strategies for studying and application of rice blast resistance genes were discussed.展开更多
DNA methylation participates in regulating the expression of coding and non-coding regions in plants. To investigate the association between DNA methylation and pathogen infection, we used whole-genome bisulfite seque...DNA methylation participates in regulating the expression of coding and non-coding regions in plants. To investigate the association between DNA methylation and pathogen infection, we used whole-genome bisulfite sequencing to survey temporal DNA methylation changes in rice after infection with the rice blast fungus Magnaporthe oryzae. In contrast to previous findings in Arabidopsis, global DNA methylation levels in rice increased slightly after rice blast infection. We identified over 38,000 differentially methylated regions(DMRs), and hypermethylated DMRs far outnumbered hypomethylated DMRs. Most DMRs were located in transposable element regions. Using transcriptome analysis, we identified 8830 differentially expressed genes(DEGs) after 1, 3, and 5 days of infection. Over one-third of DEGs, most of which were CHH-type DMRs, were associated with DMRs. Functional analysis of the CHH DMR-DEGs indicated their involvement in many important biological processes, including cell communication and response to external stimulus. The transcription of many NBS-LRR family genes was affected by changes in DNA methylation, suggesting that DNA methylation plays essential roles in the response of rice to M. oryzae infection. More broadly, the DNA methylation analysis presented here sheds light on epigenomic involvement in plant defense against fungal pathogens.展开更多
Magnaporthe oryzae,the causal agent of blast diseases,is a destructive filamentous fungus that infects many plants including most economically important food crops,rice,wheat,pearl millet and finger millet.Magnaporthe...Magnaporthe oryzae,the causal agent of blast diseases,is a destructive filamentous fungus that infects many plants including most economically important food crops,rice,wheat,pearl millet and finger millet.Magnaporthe oryzae has numerous pathotypes because of its high host-specificity in the field.The Oryza pathotype(MoO)of M.oryzae is the most devastating pathogen of rice,causing 10–30%yield loss in the world.On the other hand,the Triticum pathotype(MoT)causes blast disease in wheat,which is now a serious threat to wheat production in some South American countries,Bangladesh and Zambia.Because of low fungicide efficacy against the blast diseases and lack of availability of resistant varieties,control of rice and wheat blast diseases is difficult.Therefore,an integrated management programme should be adopted to control these two diseases in the field.Here,we introduced and summarized the classification,geographical distribution,host range,disease symptoms,biology and ecology,economic impact,and integrated pest management(IPM)programme of both rice and wheat blast diseases.展开更多
An isolate GT4028 was obtained from soil samples collected from a field in Gotsu city (Kawahira), Shimane. The use of a culture suspension and culture filtrate of this isolate significantly suppressed the spore germin...An isolate GT4028 was obtained from soil samples collected from a field in Gotsu city (Kawahira), Shimane. The use of a culture suspension and culture filtrate of this isolate significantly suppressed the spore germination in Magnaporthe oryzae. The inhibitory activity of the culture filtrate was heat-stable. The formation of rice blast lesions by M. oryzae was significantly suppressed in the presence of the culture suspension of isolate GT4028. Furthermore, mycelial growth of some plant pathogenic fungi was inhibited by the isolate in a dual culture assay. Sequence analysis of 16S rDNA region of the isolate indicated that it shared similarities with species of the genus Burkholderia. Also, isolate GT4028 could be grown even in the presence of fungicides (Blastin, Kasugamycin, and Amistar) that act against M. oryzae. These results suggest that isolate GT4028 might be a potential control agent for plant protection against diseases, such as rice blast disease.展开更多
A totatl of 116 isolates of rice blast fungus, Magnaporthe oryzae, were collected from 45 samples in different counties of Heilongjiang Province, and 20 Chinese physiological races belonging to seven groups were ident...A totatl of 116 isolates of rice blast fungus, Magnaporthe oryzae, were collected from 45 samples in different counties of Heilongjiang Province, and 20 Chinese physiological races belonging to seven groups were identified by using seven standard Chinese rice blast identifying varieties. Results showed that the dominant groups could be ranked as ZA, ZD, and ZB, with the occurrence frequencies of 47.41%, 22.41% and 15.52%, respectively. The race ZA49 was the dominant race with the occurrence frequency of 26.72%. The occurrence frequencies of the races ZD5 and ZD1 were 10.34% and 8.62%, respectively. The rising occurrence frequencies of these three dominant species were the most important reasons that causing Kongyu-131 more sensitive to rice blast. The results of virulence frequency indicated that the race harboring Pi-k, Pi-i, Pi-a gene were more susceptible to rice blast in Heilongjiang Province, and they should not be large-scale cultivated.展开更多
Eleven nitrate non-utilizing (nit) mutants were recovered from six isolates of Magnaporthe grisea cultured on MM media,amended with 60 g/L potassium chlorate, with a frequency of 1.42°. Some biological properties...Eleven nitrate non-utilizing (nit) mutants were recovered from six isolates of Magnaporthe grisea cultured on MM media,amended with 60 g/L potassium chlorate, with a frequency of 1.42°. Some biological properties, such as growth rate, growth biomass, cultural characters, conidial production, sexual reproduction ability, and pathogenicity were compared between nit mutants and their parent isolates. Results showed that all the nit mutants were resistant to chlorate. Some important biological properties such as the growth rate on YPSA, conidial production ability on TPSA, pathogenicity, had no significant differences between nit mutants and their parent isolates. Mating type didn't change, but perithecia production ability of fertile isolates changed significantly as compared with that of their parent isolates. Therefore. the nitcan be used as a genetic marker to study the genetics such as pathogenicity, fungicide resistance in Magnaporthe grisea.展开更多
Application of promoter trapping based on transformation in Magnaporthe grisea is reported in this paper. Two promoter-trapping vectors, designated as pCBGFP and pEGFPHPH, were constructed and transformed into protopl...Application of promoter trapping based on transformation in Magnaporthe grisea is reported in this paper. Two promoter-trapping vectors, designated as pCBGFP and pEGFPHPH, were constructed and transformed into protoplasts of M. grisea. A library of 1077 transformants resistant to hygromycin B was generated. Of which, 448 transformants were found to express eGFP gene in different structures of M. grisea. Three transformants grew slowly, 5 transformants decreased in conidiation and 7 transformants reduced in pathogenicity greatly among these 448 transformants. Eleven transformants were checked by genomic southern blot randomly, and 9 of which were single-copy insertions. The promoter trapping technique has been applied successfully in M. grisea and can be used as a tool for functional genomic analysis.展开更多
It is widely believed that a rice variety loses its resistance to rice blast after three to five years in commercial production due to the emergence of new Magnaporthe oryzae pathotypes(or physiological races)(Ou,1985...It is widely believed that a rice variety loses its resistance to rice blast after three to five years in commercial production due to the emergence of new Magnaporthe oryzae pathotypes(or physiological races)(Ou,1985).However,we believe that a resistant rice variety loses its resistance to M.oryzae not due to the emergence of new physiological races.We propose that pathogenic physiological races already exist in the natural environment,serving as potential physiological races.It is only because of the lack of appropriate environmental conditions,particularly the lack of a suitable rice host,on which such a race is unable to propagate widely and exists as an inferior race.However,when there is an appropriate rice host,the potential pathogenic physiological race can quickly proliferate to become the dominant physiological race,thus leading to the loss of blast resistance in resistant rice varieties.展开更多
Fungal strains isolated from the fruiting bodies of wild mushrooms were evaluated for fungicidal activity against Magnaporthe oryzae, the causal agent of the rice blast disease. Fungal isolates (n = 105) were obtained...Fungal strains isolated from the fruiting bodies of wild mushrooms were evaluated for fungicidal activity against Magnaporthe oryzae, the causal agent of the rice blast disease. Fungal isolates (n = 105) were obtained from 46 samples of wild mushrooms. Infection behaviors of M. oryzae were assessed in the presence of culture filtrates from 90 fungal isolates, of which 20 inhibited spore germination. Heat-treated culture filtrates of these isolates were classified into 3 groups according to biological activity. Blast lesion formation by M. oryzae was significantly inhibited by pretreatment with culture filtrates from 4 fungal isolates. ITS region sequence analysis indicated that these?isolates shared similarities with species of the genera Annulohypoxylon, Nigrospora, and Penicillium. Studies of symbiotic and parasitic fungi from wild mushrooms may yield potential control agents for plant diseases such as the rice blast disease.展开更多
Spore germination and appressorium formation of Magnaporthe oryzae spores was completely suppressed by an ethyl acetate extract of the culture filtrate from the H921 isolate (H921-EAE-CF). Production of antifungal sub...Spore germination and appressorium formation of Magnaporthe oryzae spores was completely suppressed by an ethyl acetate extract of the culture filtrate from the H921 isolate (H921-EAE-CF). Production of antifungal substance(s) in the H921-EAE-CF began to increase up to 3 days after isolate H921 incubation. Furthermore, heat treatment (105°C or 121°C) of H921-EAE-CF did not alter its inhibitory effect on M. oryzae spore germination compared to non-heat-treated H921-EAE-CF. Blast lesion formation inhibition by H921-EAE-CF was dose-dependent. Internal transcribed spacer (ITS) region sequence analysis indicated that this isolate shared similarities with species of the genera Trichoderma. This study suggests that H921-EAE-CF contains some antifungal substances that could be promising candidates for control of rice blast disease.展开更多
The differential hybridization teehnique has been widely used 10 identify genes that are differentially expressed. However, this approach has several drawbacks. First, the screening procedures are rather labor-intensi...The differential hybridization teehnique has been widely used 10 identify genes that are differentially expressed. However, this approach has several drawbacks. First, the screening procedures are rather labor-intensive and time. consuming. Second, the amoont of phage DNAs transferred onto the two filters may not be equivalent,展开更多
The rice blast disease has been recognized asthe most devastative disease to rice. Because of the diversity of the fungus races, frequent resistance breakdown has become a barrier totraditional breeding.
NAC transcription factors(TFs) play critical roles in plant immunity by modulating the expression of downstream genes via binding to specific cis-elements in promoters. Here, we report the function and regulatory netw...NAC transcription factors(TFs) play critical roles in plant immunity by modulating the expression of downstream genes via binding to specific cis-elements in promoters. Here, we report the function and regulatory network of a pathogenand defense phytohormone-inducible NAC TF gene, ONAC083, in rice(Oryza sativa) immunity.ONAC083 localizes to the nucleus and exhibits transcriptional activation activity that depends on its C-terminal region. Knockout of ONAC083enhances rice immunity against Magnaporthe oryzae, strengthening pathogen-induced defense responses, and boosting chitin-induced pattern-triggered immunity(PTI), whereas ONAC083 overexpression has opposite effects. We identified ONAC083-binding sites in the promoters of 82genes, and showed that ONAC083 specifically binds to a conserved element with the core sequence ACGCAA. ONAC083 activated the transcription of the genes OsRFPH2-6, OsTrx1, and OsPUP4 by directly binding to the ACGCAA element. OsRFPH2-6, encoding a RING-H2 protein with an N-terminal transmembrane region and a C-terminal typical RING domain, negatively regulated rice immunity against M. oryzae and chitin-triggered PTI. These data demonstrate that ONAC083 negatively contributes to rice immunity against M. oryzae by directly activating the transcription of OsRFPH2-6 through the ACGCAA element in its promoter. Overall, our study provides new insight into the molecular regulatory network of NAC TFs in rice immunity.展开更多
Lipid droplets are important storages in fungal conidia and can be used by plant pathogenic fungi for infection.However,the regulatory mechanism of lipid droplets formation and the utilization during fungal developmen...Lipid droplets are important storages in fungal conidia and can be used by plant pathogenic fungi for infection.However,the regulatory mechanism of lipid droplets formation and the utilization during fungal development and infection are largely unknown.Here,in Magnaporthe oryzae,we identified a lipid droplet-associated protein Nem1 that played a key role in lipid droplets biogenesis and utilization.Nem1 was highly expressed in conidia,but lowly expressed in appressoria,and its encoded protein was localized to lipid droplets.Deletion of NEM1 resulted in reduced numbers of lipid droplets and decreased content of diacylglycerol(DAG)or triacylglycerol(TAG).NEM1 was required for asexual development especially conidia production.TheΔnem1 mutant was nearly loss of virulence to host plants due to defects in appressorial penetration and invasive growth.Remarkably,Nem1 was regulated by the TOR signaling pathway and involved in the autophagy process.The Ser303 residue of Nem1 could be phosphorylated by the cAMP-PKA signaling pathway and was important for biological function of Nem1.Together,our study revealed a regulatory mechanism of lipid biogenesis and metabolism during the conidium and appressorium formation of the rice blast fungus.展开更多
基金supported by grants from the National Natural Science Foundation of China(31401692,31901960,32272513,32001976)the Natural Science Foundation of Fujian Province(2019J01766,2023J011418,2020J05177)+3 种基金Fujian Provincial Science and Technology Key Project(2022NZ030014)External Cooperation Program of Fujian Academy of Agricultural Sciences(DWHZ-2024-23)State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crop Opening Project(SKL2019005)Project of Fujian Provincial Department of Education(JAT190627)。
文摘Avirulence effectors(Avrs),encoded by plant pathogens,can be recognized by plants harboring the corresponding resistance proteins,thereby initiating effector-triggered immunity(ETI).In susceptible plants,however,Avrs can function as effectors,facilitating infection via effector-triggered susceptibility(ETS).Mechanisms of Avr-mediated ETS remain largely unexplored.Here we report that the Magnaporthe oryzae effector Avr-PikD enters rice cells via the canonical cytoplasmic secretion pathway and suppresses rice basal defense.Avr-PikD interacts with an LSD1-like transcriptional activator AKIP30 of rice,and AKIP30 is also a positive regulator of rice immunity,whereas Avr-PikD impedes its nuclear localization and suppresses its transcriptional activity.In summary,M.oryzae delivers Avr-PikD into rice cells to facilitate ETS by inhibiting AKIP30-mediated transcriptional regulation of immune response against M.oryzae.
基金Supported by Science and Technology Plan Project of Shaoguan(210804164531395)Special Fund for Modern Agricultural Industry Technology System(CARS-01-32,2022KJ105)"14thFive-Year Plan"New Discipline Team Building Project of Guangdong Academy of Agricultural Sciences(202116TD)。
文摘[Objectives]This study was conducted to screen out rice resources resistant to rice blast(Magnaporthe oryzae).[Methods]The qualitative and quantitative resistance of 1659 rice resources from 45 countries and regions to rice blast were evaluated by disease nursery in upland condition and the test of the spectrum to rice blast isolates.[Results]There were 292 entries which accounted for 17.6%showed high blast resistance(0 and 1 disease scale),68 entries(counted for 4.1%)showed resistance to blast(3 disease scale);and the number of the entries showed intermediate resistance,intermediate susceptible and susceptible were 208(with the corresponding percentage of 12.5%),471(28.4%),620(37.4%)respectively.Among the tested entries,27 entries including BG1222,BL122,BTX,IR37704-131-2-3-2,and LEBONNET had showed broad-spectrum blast resistance with the resistance frequency of higher than 90%,Quantitative resistance evaluation was conducted on some key resources,and 14 entries,of which are BR27,DRAGO,IR100,QINLIUAI、SERIBU GANTANG,YUEXIANGZHAN and so on,showed good quantitative resistances,and 8 entries had higher quantitative resistances than IR36.[Conclusions]This study provides important blast resistance resources for the local rice breeding program and has a significant value for the discovery of new blast resistance genes and its application in the blast resistance breeding.
文摘In this study, three wheat varieties were tested to determine seed germination and the incidence of Magnaporthe oryzae triticum (MoT). Among these varieties, BARI Gom 24 (Prodip) wheat seed exhibited the highest seed germination rate (93%) but also had the highest incidence (30%) of MoT. To manage blast disease in an ecofriendly manner, seven treatments were employed: T<sub>1</sub> = Control, T<sub>2</sub> = Garlic clove extracts, T<sub>3</sub> = Aloe vera leaf extracts, T<sub>4</sub> = Black cumin seed extracts, T<sub>5</sub> = Neem leaf extracts, T<sub>6</sub> = Nativo 75 WG, and T<sub>7</sub> = Provax 200 WP. The experiment was conducted using a Randomized Complete Block Design (RCBD) layout with three replications using Prodip wheat variety that exhibited highest MoT infection severity based on laboratory analysis among collected varieties. Data were collected on blast disease incidence (%), disease severity, and various growth and yield parameters of wheat. The experiment’s results indicated that among all the treatments, T<sub>7</sub> (Seed treatment with Provax 200 WP) and T<sub>5</sub> (Foliar spraying with Neem leaf extract) performed better in controlling blast disease in wheat. The lowest blast disease incidence (%) was observed with T<sub>7</sub> (Provax 200 WP), with values of 7.86, 9.86, and 10.19 recorded during the milking stage, soft dough stage, and hard dough stage of wheat, respectively. T<sub>5</sub> (Neem leaf extract) also demonstrated a statistically equivalent reduction in blast disease incidence (%). In terms of disease severity, T<sub>7</sub> (Seed treatment with Provax 200 WP) showed the lowest values of 1.03, 1.23, and 1.63 during the milking stage, soft dough stage, and hard dough stage of wheat, respectively. Foliar spraying with neem leaf extract also exhibited similar result as of Provax 200 WP regarding panicle blast severity. As a result of these findings, it can be concluded that T<sub>5</sub> (Neem leaf extract) is recommended as an ecofriendly management approach for blast disease in wheat.
文摘A mature appressorium cDNA library of rice blast fungus, Magnaporthegrisea, was constructed in a λ TriplEx2 vector by SMARTTM cDNA library containing 2.37×10^6 independent clones about 100% of which harbor foreign cDNA inserts with average size of 660 bp. Of 9 randomly selected clones, 2 expressed sequence tags (ESTs) sequences did not have homologous EST sequences of M. grisea in Gen Bank. The appressorium cDNA library is suitable for gene expression analysis and function analysis of the late stages of appressorium formation and the early stages of penetration of M. grisea.
文摘377 isolates of Magnaporthe grisea were collected from 17 provinces in China and their geographic distribution of mating types and their fertility was tested with four standard isolates, KA3 and TH12 (Mat1.1) and Guy11 and TH16 (Mat1.2) provided by CIRAD. 73 fertile isolates were tested with SCAR markers of 13 pairs of primers. Preliminary results showed that the geographic distribution of M. grisea existed among isolates collected from the same location as well as different locations and the genetic relationship between fertile isolates of the fungus in China. The existence of sexual reproduction of M. grisea was explored in the field as well.
基金The data that support the findings of this study have being submitted to GenBank and the accession numbers are JAAXMV000000000 and JAAXMU000000000.
文摘Wheat blast,caused by the fungus Magnaporthe oryzae Triticum(MoT)pathotype,is a devastating disease persistent in South America and Bangladesh.Since MoT generally fails to cause visual symptoms in wheat until the heading stage when the infection would have advanced,disease control by fungicide application solely based on the detection of visual symptoms is ineffective.To develop an accurate and sensitive method to detect MoT at the seedling and vegetative stages for disease control,we sequenced the genomes of two MoT isolates from Brazil and identified two DNA fragments,MoT-6098 and MoT-6099,that are present in the MoT genome but not in the genome of the rice-infecting Magnaporthe oryzae Oryzae(MoO)pathotype.Using polymerase chain reaction(PCR),we confirmed the specificity of the two markers in 53 MoT and MoO isolates from South America and Bangladesh.To test the efficiency of the two markers,we first established a loop-mediated isothermal amplification(LAMP)method to detect MoT at isothermal conditions,without the use of a PCR machine.Following this,we used the Cas12a protein and guide RNAs(gRNAs)to target the MoT-6098 and MoT-6099 sequences.The activated Cas12a showed indiscriminate single-stranded deoxyribonuclease(ssDNase)activity.We then combined targetdependent Cas12a ssDNase activation with recombinase polymerase amplification(RPA)and nucleic acid lateral flow immunoassay(NALFIA)to develop a method that accurately,sensitively,and cost-effectively detects MoT-specific DNA sequences in infected wheat plants.This novel technique can be easily adapted for the rapid detection of wheat blast and other important plant diseases in the field.
基金supported by the ‘948’ Project of Ministry of Agriculture, China (Grant No. 2006-G61)the Natural Science Foundation of Guangdong Province, China (Grant No. 04101156)+2 种基金the Science and Technique Project of Guangdong Province, China (Grant No. 2005B20101006 and No. 0711124900076)the Science and Technique Project of Guangzhou City, China (Grant No. 2005C12E0061)the Science Fundamental Research Foundation of Guangdong Academy of Agricultural Sciences, China.
文摘Race-specific resistance and field resistance of 30 rice blast resistance monogenic lines derived from different resources were evaluated. The spectra of resistance to 163 Magnaporthe grisea isolates collected from indica rice in Guangdong Province, China ranged from 0.6% to 89.6%. Most of the monogenic lines showed a narrow resistance spectrum and high susceptibility in rice blast area, whereas the lines with Pikh and Pi1(t) had the broad resistance spectra of 89.6% and 82.2% respectively, showing a high and stable blast resistance in fields. According to the cluster analysis of specific resistance to 163 blast isolates tested, the 30 monogenic lines were divided into 15 groups, and based on the principal factor analysis, nine kinds of race-specific resistance were identified. Pik, Piz5, Pi9 and Pish can be used as candidate resistance genes for rice breeding since their specific resistance differed from those of the backbone parents in Guangdong, China. Gene pyramiding of Pikh [or Pi1(t)], Pi9 (or Piz5) and Pish (or Pita2) will be effective to obtain broad-spectrum blast resistance in rice breeding program in Guangdong, China. The strategies for studying and application of rice blast resistance genes were discussed.
基金supported by the Natural Science Foundation of Fujian Province(2018J06006)National Key Research and Development Program of China(2016YFD0300700)+4 种基金National Natural Science Foundation of China(31770156)State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops(SKL2018006)the Program for New Century Excellent Talents of Fujian Province Universitythe Pre-eminent Youth Fund and Distinguished Young Scholars of Fujian Provincesupported by the Supercomputing Center at the College of Plant Protection of Fujian Agriculture and Forestry University。
文摘DNA methylation participates in regulating the expression of coding and non-coding regions in plants. To investigate the association between DNA methylation and pathogen infection, we used whole-genome bisulfite sequencing to survey temporal DNA methylation changes in rice after infection with the rice blast fungus Magnaporthe oryzae. In contrast to previous findings in Arabidopsis, global DNA methylation levels in rice increased slightly after rice blast infection. We identified over 38,000 differentially methylated regions(DMRs), and hypermethylated DMRs far outnumbered hypomethylated DMRs. Most DMRs were located in transposable element regions. Using transcriptome analysis, we identified 8830 differentially expressed genes(DEGs) after 1, 3, and 5 days of infection. Over one-third of DEGs, most of which were CHH-type DMRs, were associated with DMRs. Functional analysis of the CHH DMR-DEGs indicated their involvement in many important biological processes, including cell communication and response to external stimulus. The transcription of many NBS-LRR family genes was affected by changes in DNA methylation, suggesting that DNA methylation plays essential roles in the response of rice to M. oryzae infection. More broadly, the DNA methylation analysis presented here sheds light on epigenomic involvement in plant defense against fungal pathogens.
基金the National Natural Science Foundation of China(31871912,31972229 and 31772119)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-ASTIP)the grants from the Krishi Gobeshona Foundation(KGF),Bangladesh Project(TF50-C/17).
文摘Magnaporthe oryzae,the causal agent of blast diseases,is a destructive filamentous fungus that infects many plants including most economically important food crops,rice,wheat,pearl millet and finger millet.Magnaporthe oryzae has numerous pathotypes because of its high host-specificity in the field.The Oryza pathotype(MoO)of M.oryzae is the most devastating pathogen of rice,causing 10–30%yield loss in the world.On the other hand,the Triticum pathotype(MoT)causes blast disease in wheat,which is now a serious threat to wheat production in some South American countries,Bangladesh and Zambia.Because of low fungicide efficacy against the blast diseases and lack of availability of resistant varieties,control of rice and wheat blast diseases is difficult.Therefore,an integrated management programme should be adopted to control these two diseases in the field.Here,we introduced and summarized the classification,geographical distribution,host range,disease symptoms,biology and ecology,economic impact,and integrated pest management(IPM)programme of both rice and wheat blast diseases.
文摘An isolate GT4028 was obtained from soil samples collected from a field in Gotsu city (Kawahira), Shimane. The use of a culture suspension and culture filtrate of this isolate significantly suppressed the spore germination in Magnaporthe oryzae. The inhibitory activity of the culture filtrate was heat-stable. The formation of rice blast lesions by M. oryzae was significantly suppressed in the presence of the culture suspension of isolate GT4028. Furthermore, mycelial growth of some plant pathogenic fungi was inhibited by the isolate in a dual culture assay. Sequence analysis of 16S rDNA region of the isolate indicated that it shared similarities with species of the genus Burkholderia. Also, isolate GT4028 could be grown even in the presence of fungicides (Blastin, Kasugamycin, and Amistar) that act against M. oryzae. These results suggest that isolate GT4028 might be a potential control agent for plant protection against diseases, such as rice blast disease.
基金Supported by the Youth Science Funds of Heilongjiang Province (QC2010027)
文摘A totatl of 116 isolates of rice blast fungus, Magnaporthe oryzae, were collected from 45 samples in different counties of Heilongjiang Province, and 20 Chinese physiological races belonging to seven groups were identified by using seven standard Chinese rice blast identifying varieties. Results showed that the dominant groups could be ranked as ZA, ZD, and ZB, with the occurrence frequencies of 47.41%, 22.41% and 15.52%, respectively. The race ZA49 was the dominant race with the occurrence frequency of 26.72%. The occurrence frequencies of the races ZD5 and ZD1 were 10.34% and 8.62%, respectively. The rising occurrence frequencies of these three dominant species were the most important reasons that causing Kongyu-131 more sensitive to rice blast. The results of virulence frequency indicated that the race harboring Pi-k, Pi-i, Pi-a gene were more susceptible to rice blast in Heilongjiang Province, and they should not be large-scale cultivated.
文摘Eleven nitrate non-utilizing (nit) mutants were recovered from six isolates of Magnaporthe grisea cultured on MM media,amended with 60 g/L potassium chlorate, with a frequency of 1.42°. Some biological properties, such as growth rate, growth biomass, cultural characters, conidial production, sexual reproduction ability, and pathogenicity were compared between nit mutants and their parent isolates. Results showed that all the nit mutants were resistant to chlorate. Some important biological properties such as the growth rate on YPSA, conidial production ability on TPSA, pathogenicity, had no significant differences between nit mutants and their parent isolates. Mating type didn't change, but perithecia production ability of fertile isolates changed significantly as compared with that of their parent isolates. Therefore. the nitcan be used as a genetic marker to study the genetics such as pathogenicity, fungicide resistance in Magnaporthe grisea.
基金Project supported by the National Natural Science Foundation of China (Nos. 30270049 and 30470064) and the Hi-Tech Research and Development Program (863) of China (No. 2002AA245041)
文摘Application of promoter trapping based on transformation in Magnaporthe grisea is reported in this paper. Two promoter-trapping vectors, designated as pCBGFP and pEGFPHPH, were constructed and transformed into protoplasts of M. grisea. A library of 1077 transformants resistant to hygromycin B was generated. Of which, 448 transformants were found to express eGFP gene in different structures of M. grisea. Three transformants grew slowly, 5 transformants decreased in conidiation and 7 transformants reduced in pathogenicity greatly among these 448 transformants. Eleven transformants were checked by genomic southern blot randomly, and 9 of which were single-copy insertions. The promoter trapping technique has been applied successfully in M. grisea and can be used as a tool for functional genomic analysis.
基金the Key Research and Development Project of Zhejiang Province,China(Grant No.2019C02018)the National Key Research and Development Projects of China(Grant Nos.2016YFD0200801 and 2018YFD0200304)Innovation Project of Chinese Academy of Agricultural Sciences(Grant No.CAAS-ASTIP-2013-CNRRI).
文摘It is widely believed that a rice variety loses its resistance to rice blast after three to five years in commercial production due to the emergence of new Magnaporthe oryzae pathotypes(or physiological races)(Ou,1985).However,we believe that a resistant rice variety loses its resistance to M.oryzae not due to the emergence of new physiological races.We propose that pathogenic physiological races already exist in the natural environment,serving as potential physiological races.It is only because of the lack of appropriate environmental conditions,particularly the lack of a suitable rice host,on which such a race is unable to propagate widely and exists as an inferior race.However,when there is an appropriate rice host,the potential pathogenic physiological race can quickly proliferate to become the dominant physiological race,thus leading to the loss of blast resistance in resistant rice varieties.
文摘Fungal strains isolated from the fruiting bodies of wild mushrooms were evaluated for fungicidal activity against Magnaporthe oryzae, the causal agent of the rice blast disease. Fungal isolates (n = 105) were obtained from 46 samples of wild mushrooms. Infection behaviors of M. oryzae were assessed in the presence of culture filtrates from 90 fungal isolates, of which 20 inhibited spore germination. Heat-treated culture filtrates of these isolates were classified into 3 groups according to biological activity. Blast lesion formation by M. oryzae was significantly inhibited by pretreatment with culture filtrates from 4 fungal isolates. ITS region sequence analysis indicated that these?isolates shared similarities with species of the genera Annulohypoxylon, Nigrospora, and Penicillium. Studies of symbiotic and parasitic fungi from wild mushrooms may yield potential control agents for plant diseases such as the rice blast disease.
文摘Spore germination and appressorium formation of Magnaporthe oryzae spores was completely suppressed by an ethyl acetate extract of the culture filtrate from the H921 isolate (H921-EAE-CF). Production of antifungal substance(s) in the H921-EAE-CF began to increase up to 3 days after isolate H921 incubation. Furthermore, heat treatment (105°C or 121°C) of H921-EAE-CF did not alter its inhibitory effect on M. oryzae spore germination compared to non-heat-treated H921-EAE-CF. Blast lesion formation inhibition by H921-EAE-CF was dose-dependent. Internal transcribed spacer (ITS) region sequence analysis indicated that this isolate shared similarities with species of the genera Trichoderma. This study suggests that H921-EAE-CF contains some antifungal substances that could be promising candidates for control of rice blast disease.
文摘The differential hybridization teehnique has been widely used 10 identify genes that are differentially expressed. However, this approach has several drawbacks. First, the screening procedures are rather labor-intensive and time. consuming. Second, the amoont of phage DNAs transferred onto the two filters may not be equivalent,
文摘The rice blast disease has been recognized asthe most devastative disease to rice. Because of the diversity of the fungus races, frequent resistance breakdown has become a barrier totraditional breeding.
基金supported by grants from the National Natural Science Foundation of China (31871945, 32072403)the National Key Research and Development Program of China (2016YFD0100600)。
文摘NAC transcription factors(TFs) play critical roles in plant immunity by modulating the expression of downstream genes via binding to specific cis-elements in promoters. Here, we report the function and regulatory network of a pathogenand defense phytohormone-inducible NAC TF gene, ONAC083, in rice(Oryza sativa) immunity.ONAC083 localizes to the nucleus and exhibits transcriptional activation activity that depends on its C-terminal region. Knockout of ONAC083enhances rice immunity against Magnaporthe oryzae, strengthening pathogen-induced defense responses, and boosting chitin-induced pattern-triggered immunity(PTI), whereas ONAC083 overexpression has opposite effects. We identified ONAC083-binding sites in the promoters of 82genes, and showed that ONAC083 specifically binds to a conserved element with the core sequence ACGCAA. ONAC083 activated the transcription of the genes OsRFPH2-6, OsTrx1, and OsPUP4 by directly binding to the ACGCAA element. OsRFPH2-6, encoding a RING-H2 protein with an N-terminal transmembrane region and a C-terminal typical RING domain, negatively regulated rice immunity against M. oryzae and chitin-triggered PTI. These data demonstrate that ONAC083 negatively contributes to rice immunity against M. oryzae by directly activating the transcription of OsRFPH2-6 through the ACGCAA element in its promoter. Overall, our study provides new insight into the molecular regulatory network of NAC TFs in rice immunity.
基金supported by the National Natural Science Foundation of China(Grants 32072365 and 32272476)the Open Research Fund of the State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China(SKL-KF202216).
文摘Lipid droplets are important storages in fungal conidia and can be used by plant pathogenic fungi for infection.However,the regulatory mechanism of lipid droplets formation and the utilization during fungal development and infection are largely unknown.Here,in Magnaporthe oryzae,we identified a lipid droplet-associated protein Nem1 that played a key role in lipid droplets biogenesis and utilization.Nem1 was highly expressed in conidia,but lowly expressed in appressoria,and its encoded protein was localized to lipid droplets.Deletion of NEM1 resulted in reduced numbers of lipid droplets and decreased content of diacylglycerol(DAG)or triacylglycerol(TAG).NEM1 was required for asexual development especially conidia production.TheΔnem1 mutant was nearly loss of virulence to host plants due to defects in appressorial penetration and invasive growth.Remarkably,Nem1 was regulated by the TOR signaling pathway and involved in the autophagy process.The Ser303 residue of Nem1 could be phosphorylated by the cAMP-PKA signaling pathway and was important for biological function of Nem1.Together,our study revealed a regulatory mechanism of lipid biogenesis and metabolism during the conidium and appressorium formation of the rice blast fungus.
