Rice sheath blight, caused by Rhizoctonia solani AG1-IA, is a major disease in rice-growing areas worldwide. Effectors of phytopathogenic fungi play important roles during the infection process of fungal pathogens ont...Rice sheath blight, caused by Rhizoctonia solani AG1-IA, is a major disease in rice-growing areas worldwide. Effectors of phytopathogenic fungi play important roles during the infection process of fungal pathogens onto their host plants. However, the molecular mechanisms by which R. solani effectors regulate rice immunity are not well understood. Through prediction, 78 candidate effector molecules were identified. Using the tobacco rattle virus-host induced gene silencing(TRV-HIGS) system, 45 RNAi constructs of effector genes were infiltrated into Nicotiana benthamiana leaves. The results revealed that eight of these constructs resulted in a significant reduction in necrosis caused by infection with the AG1-IA strain GD-118. Additionally, stable rice transformants carrying the double-stranded RNA construct for one of the effector genes, AGLIP1, were generated to further verify the function of this gene. The suppression of the AGLIP1 gene increased the resistance of both N. benthamiana and rice against GD-118, and also affected the growth rate of GD-118, indicating that AGLIP1 is a key pathogenic factor. Small RNA sequencing showed that the HIGS vectors were processed into si RNAs within the plants and then translocated to the fungi, leading to the silencing of the target genes. As a result, AGLIP1 might be an excellent candidate for HIGS, thereby enhancing crop resistance against the pathogen and contributing to the control of R. solani infection.展开更多
Pear is a fruit crop of worldwide importance and cold storage is an integral part of the production and distribution of pears.An uncharacterized fungal disease has been observed on‘Huangguan’pear fruit during cold s...Pear is a fruit crop of worldwide importance and cold storage is an integral part of the production and distribution of pears.An uncharacterized fungal disease has been observed on‘Huangguan’pear fruit during cold storage in Hebei Province.The fungus was consistently isolated from diseased fruit by routine tissue separation method,and shown to be the causal agent according to Koch postulates.Based on its morphology,molecular characteristics,pathogenicity and ITS sequence,the fungus was identified as Rhizoctonia solani.This study recorded postharvest fruit rot caused by Rhizoctonia solani on pear fruit in China.展开更多
分别基于7个烟草靶斑病菌株基因组中ITS-5.8S r DNA序列设计引物和探针;并对引物及探针特异性进行验证;建立检测体系并对接种烟草靶斑病菌的叶片和土壤中的烟草靶斑病菌进行检测。结果表明:所设计的引物及探针对R.solani AG-3具有特异性...分别基于7个烟草靶斑病菌株基因组中ITS-5.8S r DNA序列设计引物和探针;并对引物及探针特异性进行验证;建立检测体系并对接种烟草靶斑病菌的叶片和土壤中的烟草靶斑病菌进行检测。结果表明:所设计的引物及探针对R.solani AG-3具有特异性,检测体系可以检测出烟草叶片及土壤样品中的烟草靶斑病菌。接种烟草叶片的检测表明,接种后6h就可检测到强致病力菌株YC-9,12h后能检测到弱致病力菌株LF-2;获得了烟草靶斑病菌DNA质量的对数与添加菌丝量的对数之间的回归曲线方程,对不同月份土壤样品的测定结果表明烟草靶斑病菌在土壤中呈周年动态变化趋势。展开更多
Rice sheath blight disease （ShB）, caused by Rhizoctonia solani, gives rise to significant grain yield losses. The present study evaluated the efficacy of Integral, the commercial liquid formulation of Bacillus subti...Rice sheath blight disease （ShB）, caused by Rhizoctonia solani, gives rise to significant grain yield losses. The present study evaluated the efficacy of Integral, the commercial liquid formulation of Bacillus subtilis strain MBI 600, against rice ShB and for plant growth promotion. In greenhouse studies, four log concentrations of Integral （from 2.2×10^6 to 2.2×10^9 cfu/mL） were used as seed treatment （ST）. After 25 d, seedlings were dipped （SD） into Integral prior to transplanting. At 30 d after transplanting （DAT）, leaf sheaths were inoculated with immature sclerotia of the pathogen. At 45 DAT, a foliar spray （FS） with Integral was applied to some treatments. The fungicide control was 50% carbendazim at 1.0 g/L, and a nontreated control was also included. Overall, there were 10 treatments, each with five replications. ShB severity was rated at 52 DAT, and seedling height and number of tillers per plant were rated at 60 DAT. In 2009, two field trials evaluated Integral at 2.2×10^8 and 2.2×10^9 cfu/mL. Integral was applied as ST, and seedlings were produced in a nursery bed. After 32 d, seedlings were treated with Integral as SD and transplanted into 10 m^2 blocks. Foliar sprays were given at 45 and 60 DAT. There were seven treatments, each with eight replications arranged as a factorial randomized complete block design. At 20 DAT, the plots were broadcast inoculated with R. solani produced on rice grains. Seedling height before transplanting, ShB severity at 90 DAT, and grain yield at harvest were recorded. Integral at 2.2×10^9 cfu/mL provided significant increase of seedling heights over other treatments under greenhouse conditions. The Integral treatments of ST ＋ SD ＋ FS at 2.2×10^9 cfu/mL significantly suppressed ShB over other treatments. In field studies, Integral provided significant increase of seedling height in nursery, and number of tillers per plant, compared with the control. ShB severity was significantly suppressed with higher concentrations of Integral compared to lower concentrations. Grain yield were the highest at an Integral concentration of 2.2×10^9 cfu/mL. Overall, Integral significantly reduced ShB severity, enhanced seedling growth, number of tillers per plant and grain yield as ST ＋ SD ＋ FS at the concentration of 2.2×10^9 cfu/mL under the conditions evaluated.展开更多
The inhibition of lanthanum (La) to mycelial growth and three disease-related enzymes of Rhizoctonia solani were studied. The results showed that lanthanum inhibits the growth of Rhizoctonia solani strongly. EC_(50) a...The inhibition of lanthanum (La) to mycelial growth and three disease-related enzymes of Rhizoctonia solani were studied. The results showed that lanthanum inhibits the growth of Rhizoctonia solani strongly. EC_(50) and EC_(95) of La were 171.9 and 667.7 mg·L^(-1) measured in solid culture media respectively, while 111.4 and 500.7 mg·L^(-1) measured in liquid culture media respectively. Lanthanum also has activating effects on disease-related enzymes of the fungus such as pectinase, protease and cellulase. However, the quantity or the activity of the total enzymes decreases significantly because of the strong blockage of mycelial growth when the La_2O_3 concentration is over 50 mg·L^(-1), and the virulence of pathogen decreases as well.展开更多
Sheath blight （SB） caused by Rhizoctonia solani is one of the three major diseases of rice, and now has become the most severe disease causing rice yield loss in China. Breeding and use of varieties resistant to SB ...Sheath blight （SB） caused by Rhizoctonia solani is one of the three major diseases of rice, and now has become the most severe disease causing rice yield loss in China. Breeding and use of varieties resistant to SB is crucial in controlling the disease, but the advances achieved have been limited due to the tack of highly SB-resistant rice germplasm. Genetic analysis revealed that the SB resistance in rice was a typical quantitative trait controlled by multi-genes. Although many QTLs conferring resistance to SB have been identified in recent years, most of the QTLs only showed small effects and few of them have been evaluated for utilization potential. Many R. solani-resistant resources have been found in wild rice species, microorganisms and other plant species. It is already known that the SB-resistance could be improved in transgenic rice plants by genetic transformation. This paper reviewed the genetic mapping of loci associated with resistance to rice SB, the evaluation of the potential of resistance QTLs, and the resistant resources found in various organisms besides rice. To develop SB-resistant rice varieties, it is important to develop and explore new resistant rice germplasms, fine map and evaluate resistance QTLs, and also to pay attention to various bio-resources showing resistance to R. solani.展开更多
Eight compounds were isolated from the fermentation cultures of rice sheath blight pathogen Rhizoctonia solani Kuhn. They were identified as ergosterol （1）, 6β-hydroxysitostenone （2）, sitostenone （3）, m-hydroxy...Eight compounds were isolated from the fermentation cultures of rice sheath blight pathogen Rhizoctonia solani Kuhn. They were identified as ergosterol （1）, 6β-hydroxysitostenone （2）, sitostenone （3）, m-hydroxyphenylacetic acid （4）, methyl m-hydroxyphenylacetate （5）, m-hydroxymethylphenyl pentanoate （6）, （Z）-3-methylpent-2-en-1,5-dioic acid （7） and 3-methoxyfuran-2-carboxylic acid （8） by means of physicochemical and spectroscopic analysis. Among them, 2, 3, 5-8 were isolated from R. solani for the first time. All the compounds were evaluated for their biological activities. 4-6 and 8 showed their inhibitory activities on the radical and germ elongation of rice seeds. 1,4 and 7 showed moderate antibacterial activity to some bacteria. 4, 7 and 8 exhibited weak inhibitory activities on spore germination of Magnaporthe oryzae. 8 showed moderate antioxidant activity with the 1,1-diphenyl-2-picryhydrazyl （DPPH） andβ-carotene-linoleic acid assays. This is the first time to reveal compounds 5, 6 and 8 from rice sheath blight pathogen R. solani to have in vitro phytotoxic activity.展开更多
To construct the T-DNA insertional mutagenesis transformation system for rice sheath blight pathogen Rhizoctonia solani AG-1 IA,the virulent isolate GD118 of this pathogen was selected as an initial isolate for transf...To construct the T-DNA insertional mutagenesis transformation system for rice sheath blight pathogen Rhizoctonia solani AG-1 IA,the virulent isolate GD118 of this pathogen was selected as an initial isolate for transformation.The conditions for transformation of isolate GD118 were optimized in five aspects,i.e.pre-induction time,co-culture time,acetosyringone(AS) concentration at the co-culture phase,co-culture temperature and pH value of induction solid medium(ISM) at the co-culture phase.Finally,a system of Agrobacterium tumefaciens-mediated transformation(ATMT) for R.solani AG-1 IA was established successfully.The optimal conditions for this ATMT system were as follows:the concentration of hygromycin B at 30 μg/mL for transformant screening,8 h of pre-induction,20 h of co-culture,200 μmol/L of AS in ISM,co-culture at 25 ℃ and pH 5.6 to 5.8 of ISM at the co-culture phase.The transformants still displayed high resistance to hygromycin B after subculture for five generations.A total of 10 randomly selected transformants were used for PCR verification using the specific primers designed for the hph gene,and the results revealed that an expected band of 500 bp was amplified from all of the 10 transformants.Moreover,PCR amplification for these 10 transformants was carried out using specific primers designed for the Vir gene of A.tumefaciens,with four strains of A.tumefaciens as positive controls for eliminating the false-positive caused by the contamination of A.tumefaciens.An expected band of 730 bp was amplified from the four strains of A.tumefaciens,whereas no corresponding DNA band could be amplified from the 10 transformants.The results of the two PCR amplifications clearly showed that T-DNA was indeed inserted into the genome of target isolate GD118.展开更多
基金supported by the Henan Province Science and Technology Research Project, China (Grant No. 242102110232)the National Natural Science Foundation of China (Grant No. 31801677)the Major Program of Guangdong Basic and Applied Basic Research, China (Grant No. 2019B030302006)。
文摘Rice sheath blight, caused by Rhizoctonia solani AG1-IA, is a major disease in rice-growing areas worldwide. Effectors of phytopathogenic fungi play important roles during the infection process of fungal pathogens onto their host plants. However, the molecular mechanisms by which R. solani effectors regulate rice immunity are not well understood. Through prediction, 78 candidate effector molecules were identified. Using the tobacco rattle virus-host induced gene silencing(TRV-HIGS) system, 45 RNAi constructs of effector genes were infiltrated into Nicotiana benthamiana leaves. The results revealed that eight of these constructs resulted in a significant reduction in necrosis caused by infection with the AG1-IA strain GD-118. Additionally, stable rice transformants carrying the double-stranded RNA construct for one of the effector genes, AGLIP1, were generated to further verify the function of this gene. The suppression of the AGLIP1 gene increased the resistance of both N. benthamiana and rice against GD-118, and also affected the growth rate of GD-118, indicating that AGLIP1 is a key pathogenic factor. Small RNA sequencing showed that the HIGS vectors were processed into si RNAs within the plants and then translocated to the fungi, leading to the silencing of the target genes. As a result, AGLIP1 might be an excellent candidate for HIGS, thereby enhancing crop resistance against the pathogen and contributing to the control of R. solani infection.
基金supported by the National Key R&D Program of China(Grant No.2016YFD0400903–06)the earmarked fund for China Agriculture Research System(Grant No.CARS-29–19)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences。
文摘Pear is a fruit crop of worldwide importance and cold storage is an integral part of the production and distribution of pears.An uncharacterized fungal disease has been observed on‘Huangguan’pear fruit during cold storage in Hebei Province.The fungus was consistently isolated from diseased fruit by routine tissue separation method,and shown to be the causal agent according to Koch postulates.Based on its morphology,molecular characteristics,pathogenicity and ITS sequence,the fungus was identified as Rhizoctonia solani.This study recorded postharvest fruit rot caused by Rhizoctonia solani on pear fruit in China.
文摘分别基于7个烟草靶斑病菌株基因组中ITS-5.8S r DNA序列设计引物和探针;并对引物及探针特异性进行验证;建立检测体系并对接种烟草靶斑病菌的叶片和土壤中的烟草靶斑病菌进行检测。结果表明:所设计的引物及探针对R.solani AG-3具有特异性,检测体系可以检测出烟草叶片及土壤样品中的烟草靶斑病菌。接种烟草叶片的检测表明,接种后6h就可检测到强致病力菌株YC-9,12h后能检测到弱致病力菌株LF-2;获得了烟草靶斑病菌DNA质量的对数与添加菌丝量的对数之间的回归曲线方程,对不同月份土壤样品的测定结果表明烟草靶斑病菌在土壤中呈周年动态变化趋势。
文摘Rice sheath blight disease （ShB）, caused by Rhizoctonia solani, gives rise to significant grain yield losses. The present study evaluated the efficacy of Integral, the commercial liquid formulation of Bacillus subtilis strain MBI 600, against rice ShB and for plant growth promotion. In greenhouse studies, four log concentrations of Integral （from 2.2×10^6 to 2.2×10^9 cfu/mL） were used as seed treatment （ST）. After 25 d, seedlings were dipped （SD） into Integral prior to transplanting. At 30 d after transplanting （DAT）, leaf sheaths were inoculated with immature sclerotia of the pathogen. At 45 DAT, a foliar spray （FS） with Integral was applied to some treatments. The fungicide control was 50% carbendazim at 1.0 g/L, and a nontreated control was also included. Overall, there were 10 treatments, each with five replications. ShB severity was rated at 52 DAT, and seedling height and number of tillers per plant were rated at 60 DAT. In 2009, two field trials evaluated Integral at 2.2×10^8 and 2.2×10^9 cfu/mL. Integral was applied as ST, and seedlings were produced in a nursery bed. After 32 d, seedlings were treated with Integral as SD and transplanted into 10 m^2 blocks. Foliar sprays were given at 45 and 60 DAT. There were seven treatments, each with eight replications arranged as a factorial randomized complete block design. At 20 DAT, the plots were broadcast inoculated with R. solani produced on rice grains. Seedling height before transplanting, ShB severity at 90 DAT, and grain yield at harvest were recorded. Integral at 2.2×10^9 cfu/mL provided significant increase of seedling heights over other treatments under greenhouse conditions. The Integral treatments of ST ＋ SD ＋ FS at 2.2×10^9 cfu/mL significantly suppressed ShB over other treatments. In field studies, Integral provided significant increase of seedling height in nursery, and number of tillers per plant, compared with the control. ShB severity was significantly suppressed with higher concentrations of Integral compared to lower concentrations. Grain yield were the highest at an Integral concentration of 2.2×10^9 cfu/mL. Overall, Integral significantly reduced ShB severity, enhanced seedling growth, number of tillers per plant and grain yield as ST ＋ SD ＋ FS at the concentration of 2.2×10^9 cfu/mL under the conditions evaluated.
文摘The inhibition of lanthanum (La) to mycelial growth and three disease-related enzymes of Rhizoctonia solani were studied. The results showed that lanthanum inhibits the growth of Rhizoctonia solani strongly. EC_(50) and EC_(95) of La were 171.9 and 667.7 mg·L^(-1) measured in solid culture media respectively, while 111.4 and 500.7 mg·L^(-1) measured in liquid culture media respectively. Lanthanum also has activating effects on disease-related enzymes of the fungus such as pectinase, protease and cellulase. However, the quantity or the activity of the total enzymes decreases significantly because of the strong blockage of mycelial growth when the La_2O_3 concentration is over 50 mg·L^(-1), and the virulence of pathogen decreases as well.
基金supported by the grant from the Technological Key Project of Ministry of Agriculture of China on New Transgenic Variety Breeding (Grant No. 2008ZX08001-02)
文摘Sheath blight （SB） caused by Rhizoctonia solani is one of the three major diseases of rice, and now has become the most severe disease causing rice yield loss in China. Breeding and use of varieties resistant to SB is crucial in controlling the disease, but the advances achieved have been limited due to the tack of highly SB-resistant rice germplasm. Genetic analysis revealed that the SB resistance in rice was a typical quantitative trait controlled by multi-genes. Although many QTLs conferring resistance to SB have been identified in recent years, most of the QTLs only showed small effects and few of them have been evaluated for utilization potential. Many R. solani-resistant resources have been found in wild rice species, microorganisms and other plant species. It is already known that the SB-resistance could be improved in transgenic rice plants by genetic transformation. This paper reviewed the genetic mapping of loci associated with resistance to rice SB, the evaluation of the potential of resistance QTLs, and the resistant resources found in various organisms besides rice. To develop SB-resistant rice varieties, it is important to develop and explore new resistant rice germplasms, fine map and evaluate resistance QTLs, and also to pay attention to various bio-resources showing resistance to R. solani.
基金co-financed by the grants from the National Basic Resea rch Program of China (2013GB127800)the National Natural Science Foundation of China (31271996)
文摘Eight compounds were isolated from the fermentation cultures of rice sheath blight pathogen Rhizoctonia solani Kuhn. They were identified as ergosterol （1）, 6β-hydroxysitostenone （2）, sitostenone （3）, m-hydroxyphenylacetic acid （4）, methyl m-hydroxyphenylacetate （5）, m-hydroxymethylphenyl pentanoate （6）, （Z）-3-methylpent-2-en-1,5-dioic acid （7） and 3-methoxyfuran-2-carboxylic acid （8） by means of physicochemical and spectroscopic analysis. Among them, 2, 3, 5-8 were isolated from R. solani for the first time. All the compounds were evaluated for their biological activities. 4-6 and 8 showed their inhibitory activities on the radical and germ elongation of rice seeds. 1,4 and 7 showed moderate antibacterial activity to some bacteria. 4, 7 and 8 exhibited weak inhibitory activities on spore germination of Magnaporthe oryzae. 8 showed moderate antioxidant activity with the 1,1-diphenyl-2-picryhydrazyl （DPPH） andβ-carotene-linoleic acid assays. This is the first time to reveal compounds 5, 6 and 8 from rice sheath blight pathogen R. solani to have in vitro phytotoxic activity.
基金supported by a ‘Special Fund for Agro-scientific Research in the Public Interest’ from the Ministry of Agriculture of China(Grant No.nyhyzx3-16)
文摘To construct the T-DNA insertional mutagenesis transformation system for rice sheath blight pathogen Rhizoctonia solani AG-1 IA,the virulent isolate GD118 of this pathogen was selected as an initial isolate for transformation.The conditions for transformation of isolate GD118 were optimized in five aspects,i.e.pre-induction time,co-culture time,acetosyringone(AS) concentration at the co-culture phase,co-culture temperature and pH value of induction solid medium(ISM) at the co-culture phase.Finally,a system of Agrobacterium tumefaciens-mediated transformation(ATMT) for R.solani AG-1 IA was established successfully.The optimal conditions for this ATMT system were as follows:the concentration of hygromycin B at 30 μg/mL for transformant screening,8 h of pre-induction,20 h of co-culture,200 μmol/L of AS in ISM,co-culture at 25 ℃ and pH 5.6 to 5.8 of ISM at the co-culture phase.The transformants still displayed high resistance to hygromycin B after subculture for five generations.A total of 10 randomly selected transformants were used for PCR verification using the specific primers designed for the hph gene,and the results revealed that an expected band of 500 bp was amplified from all of the 10 transformants.Moreover,PCR amplification for these 10 transformants was carried out using specific primers designed for the Vir gene of A.tumefaciens,with four strains of A.tumefaciens as positive controls for eliminating the false-positive caused by the contamination of A.tumefaciens.An expected band of 730 bp was amplified from the four strains of A.tumefaciens,whereas no corresponding DNA band could be amplified from the 10 transformants.The results of the two PCR amplifications clearly showed that T-DNA was indeed inserted into the genome of target isolate GD118.
