Among the viruses Cucumber mosaic virus (CMV) has been rated worldwide as one of the five most important viruses infecting vegetable species. CMV is a tripartite virus with high sequence variability, classified into t...Among the viruses Cucumber mosaic virus (CMV) has been rated worldwide as one of the five most important viruses infecting vegetable species. CMV is a tripartite virus with high sequence variability, classified into three subgroups with 80% to 97% identical nucleotides in their coat protein. Due to the absence of natural resistance CMV is the plant virus with longest history in genetic engineering using pathogen induced approaches. However, the transformation and regeneration for some very important crops like chili is difficult. Therefore it will be an advantage to screen in model plants for gene constructs which might be independent of the target of final transformation and other parameters having an influence on the efficiency of a biotechnological approach. In our study we compared the resistance for all combinations of five different antiviral constructs, two different transformation vectors and two model host plants. From these approaches we identified the most effective construct which might also be applicable to transform eventually chili plants.展开更多
[ Objective] The paper was to study the resistance of barnyard grass to quinclorac in double-harvest rice area in Hunan Province of China. [ Method ] Using whole-plant determination method, the resistance of 36 biotyp...[ Objective] The paper was to study the resistance of barnyard grass to quinclorac in double-harvest rice area in Hunan Province of China. [ Method ] Using whole-plant determination method, the resistance of 36 biotypes of barnyard grass to quinclorac in double-harvest rice area in Hunan Province was studied. [ Result ] The biotype collected from Lijiaping Town of Shaoyang County was the most susceptible barnyard grass to quinclerac, and the EC50 value was 70. 253 1 g a. i./hm2 , so it was recognized as the sensitive biotype. The biotypes collected from Caowei Town of Yuanjiang County （ HN3 and HN4 from direct seeding rice fields） and Zhongyukou Town of Nanxian County presented very high resistance to quinclorac ; the ECho values were 1 999. 122 5, 1 664.544 6 and 1 022.739 5 g a. i./hm2, and their resistance indexes reached 28.46, 23.69 and 14.56, respectively, indicating that these throe biotypes hod high level of resistance to quinclo- rac. EC50 values of about 22 biotypes were ranged from 144. 837 3 to 408.289 4 g a. i./hm2, and resistance indexes were ranged from 2.06 to 5.81, indicating that these biotypes hod produced resistance to quinclorac. EC50 values of other 10 biotypes were ranged from 98.032 6 to 138. 660 9 g a. i./hm2, and the resistance in- dexes were ranged from 1.40 to 1.97, indicating that the sensitivity of barnyard grass in these places was decreasing. [ Conclusion] The paper provided guidance for scientific and rational use of quinclorac in paddy fields.展开更多
Pepino mosaic virus(PepMV)causes severe disease in tomato and other Solanaceous crops around globe.To effectively study and manage this viral disease,researchers need new,sensitive,and high-throughput approaches for v...Pepino mosaic virus(PepMV)causes severe disease in tomato and other Solanaceous crops around globe.To effectively study and manage this viral disease,researchers need new,sensitive,and high-throughput approaches for viral detection.In this study,we purified PepMV particles from the infected Nicotiana benthamiana plants and used virions to immunize BALB/c mice to prepare hybridomas secreting anti-PepMV monoclonal antibodies(mAbs).A panel of highly specific and sensitive murine mAbs(15B2,8H6,23D11,20D9,3A6,and 8E3)could be produced through cell fusion,antibody selection,and cell cloning.Using the mAbs as the detection antibodies,we established double antibody sandwich enzyme-linked immunosorbent assay(DAS-ELISA),Dot-ELISA,and Tissue print-ELISA for detecting PepMV infection in tomato plants.Resulting data on sensitivity analysis assays showed that both DAS-ELISA and Dot-ELISA can efficiently monitor the virus in PepMV-infected tissue crude extracts when diluted at 1:1310720 and 1:20480(weight/volume ratio(w/v),g/mL),respectively.Among the three methods developed,the Tissue print-ELISA was found to be the most practical detection technique.Survey results from field samples by the established serological approaches were verified by reverse transcription polymerase chain reaction(RT-PCR)and DNA sequencing,dem on strati ng all three serological methods are reliable and effective for monitoring PepMV.An ti-PepMV mAbs and the newly developed DAS-ELISA,Dot-ELISA,and Tissue print-ELISA can benefit PepMV detection and field epidemiological study,and management of this viral disease,which is already widespread in tomato plants in Yunnan Province of China.展开更多
One sequence tagged site marker Subl-1 and twice submergence stress method were used in selection of submergence tolerant homozygous line from Sub-lBS, a submergence tolerant, bentazon sensitive and photoperiod-sensit...One sequence tagged site marker Subl-1 and twice submergence stress method were used in selection of submergence tolerant homozygous line from Sub-lBS, a submergence tolerant, bentazon sensitive and photoperiod-sensitive and/or thermo-sensitive genic male sterile line that developed by our laboratory. The results revealed that the original Sub-lBS was heterozygous in SublA-1 locus even though it was identical in almost all of agronomical traits and the segregation of SublA-1 was in accordance with Mendelian law based on chi-square test. And then the original Sub-IBS was divided into two groups： one was ofSublA-1 introgression and the other was not; and the two groups were tested by twice submergence stress method. After the first submergence stress that lasted for 12 d, the average plant heights were significant difference at the 1% level between the two groups. After recovery for 10 d, the second submergence stress sustained for 18 d was carried on; and the group with SublA-1 gene was found apparently tolerant than the other group in submergence tolerance.展开更多
Soybean chlorotic mottle virus(SbCMV)was first detected from soybean plants in Jiangxi Province of China by high throughput sequencing and was confirmed by PCR.The complete nucleotide sequence of NC113 was determined ...Soybean chlorotic mottle virus(SbCMV)was first detected from soybean plants in Jiangxi Province of China by high throughput sequencing and was confirmed by PCR.The complete nucleotide sequence of NC113 was determined to be 8210 nucleotides,and shared the highest similarity(91.7%)with sequences of SbCMV that was only reported in Japan.It encodes nine putative open reading frames(ORFs Ia,Ib and Ⅱ-Ⅷ),and contains a large intergenic region located at nucleotide 5976-6512 between ORFs VI and VII.Sequence analysis and phylogenetic tree indicated that NC113 is an isolate of SbCMV,and is more related to the soymoviruses Blueberry red ringspot virus(BRRSV),Peanut chlorotic streak virus(PCSV)and Cestrum yellow leaf curling virus(CmYLCV)than to other representative members in the Caulimoviridae family.Field survey of 472 legume plants from Jiangxi and Zhejiang provinces showed SbCMV was only detected from soybean in Nanchang City with a low incidence rate.This is the first report of Soybean chlorotic mottle virus identified in China.展开更多
In this study, the characteristic of three transformants named as B1C106-1, B1C106-2, and B1C106-3 were studied that carried three innate resistant genes Bph14, Bph15, and Xa23, and two enthetic resistant genes CrylCa...In this study, the characteristic of three transformants named as B1C106-1, B1C106-2, and B1C106-3 were studied that carried three innate resistant genes Bph14, Bph15, and Xa23, and two enthetic resistant genes CrylCa# and Bar. The five resistant genes were all verified by PCR and the two enthetic genes were identified in single copy insertion by Southern blot. At tillering stage, the CrylC and PAT （phosphinothricin acetyl transferase） protein contents in leaf, sheath, and stem of T2 generation were in the similar pattern： leaf〉stem〉sheath, and showed significant difference （P〈0.01） among three organs. The average contents of CrylC protein in plant of B1C106-1, B1C106-2, and BIC106-3 were 12.95, 6.57, and 11.30 μg g-1, respectively, and showed significant difference （P〈0.01） among them. However, the average contents of PAT in plant of B1C106-1, B1C106-2, and B1C106-3 were 28.54, 27.66, and 28.02 pg g-1, respectively, and there were no significant difference among three transformants. The glufosinate tolerable concentration of three transformants of T3 generation reached at least 6 g L-1, and the mortality of rice leaf rollers were above 97.4% in 5 days after being fed with fresh transformants＇ leaves. The CrylC protein toxicity was also assessed by silkworms, and the mortality of silkworms feeding mulberry leaves smeared with Cry1C protein extracts of leaves of B1 C106-1, BIC106-2, and B1 C106-3 were 90, 67.8, and 87.8%, respectively, that were positive correlation （r=0.993） with CrylC protein contents in plant of three transformants. The three transformants also maintained high resistance to brown planthopper and bacterial blight as the original version. The above results indicate the tetra-resistant rice germplasm was well-developed by pyramiding innate and enthetic resistant genes in an elite line to provide with resistances of glufosinate, rice leaf roller, brown planthopper, and bacterial blight.展开更多
[Objectives]A codominance functional marker of the broad-spectrum bacterial blight resistance gene,Xa7,of rice was identified for accurate detection,generation tracking,and differentiation between homozygous and hemiz...[Objectives]A codominance functional marker of the broad-spectrum bacterial blight resistance gene,Xa7,of rice was identified for accurate detection,generation tracking,and differentiation between homozygous and hemizygous genotypes of the gene.[Methods]A potential functional marker containing four primers was designed using Premier 5 software and based on the differences on the sequences of Xa7,xa7,and allele-free genomes.The molecular distinctness of the marker in different materials was verified by PCR.Three crossbreed lines of Xa7 and their parents were inoculated with seven bacterial blight strains at the booting stage to examine the affected agronomic traits at maturation.[Results]The homozygous R084 of Xa7 could be amplified into a 91 bp band and the Nip free of allele with a 153 bp band,while the heterozygote Nip/R084,91 bp and 153 bp bands.The candidate codominance marker,Xa7fun,amplified fragments that matched the predicted target bands.No 91 bp fragment was amplified from 18 germplasms of varied types,indicating a lack of Xa7 in them.Whereas Ry1,Ry2 and Ry3 had a 91 bp band,suggesting the inclusion of homozygous Xa7.Under an elevated temperature,Huazhan responded to the seven bacterial blight pathogens as highly susceptible(HS),intermediate susceptible(MS),or susceptible(S);R084 to six of the seven pathogens(HNA1-4,FuJ,GDA2,GD1358,PX086,and YN24)as highly resistant(HR),intermediate resistant(MR)or resistant(R);Ry-1 to five pathogens(GDA2,HNA1-4,FuJ,GD1358,and YN24)as HR or MR;Ry-2 to five pathogens(GDA2,GD1358,HNA1-4,PXO86,and YN24)as HR or R;and Ry-3 to 6 pathogens(HNA1-4,FuJ,GDA2,GD1358,PXO86,and YN24)as HR or MR.Therefore,the infiltration of Xa7 in the improved crossbred lines RY-1,RY-2,and RY-3 significantly accentuated the blight resistance of Huazhan.[Conclusions]Homozygous or hemizygous Xa7 could be accurately differentiated by the currently identified codominance functional marker Xa7 fun.The Xa7 introgression did not significantly alter the critical agronomic traits in the hybridization from generation to generation and could be safely applied in breeding rice varieties with bacterial blight resistance.展开更多
文摘Among the viruses Cucumber mosaic virus (CMV) has been rated worldwide as one of the five most important viruses infecting vegetable species. CMV is a tripartite virus with high sequence variability, classified into three subgroups with 80% to 97% identical nucleotides in their coat protein. Due to the absence of natural resistance CMV is the plant virus with longest history in genetic engineering using pathogen induced approaches. However, the transformation and regeneration for some very important crops like chili is difficult. Therefore it will be an advantage to screen in model plants for gene constructs which might be independent of the target of final transformation and other parameters having an influence on the efficiency of a biotechnological approach. In our study we compared the resistance for all combinations of five different antiviral constructs, two different transformation vectors and two model host plants. From these approaches we identified the most effective construct which might also be applicable to transform eventually chili plants.
基金Supported by National Natural Science Foundation of China(31201530)Special Fund for Agro-scientific Research in the Public Interest(201303031)
文摘[ Objective] The paper was to study the resistance of barnyard grass to quinclorac in double-harvest rice area in Hunan Province of China. [ Method ] Using whole-plant determination method, the resistance of 36 biotypes of barnyard grass to quinclorac in double-harvest rice area in Hunan Province was studied. [ Result ] The biotype collected from Lijiaping Town of Shaoyang County was the most susceptible barnyard grass to quinclerac, and the EC50 value was 70. 253 1 g a. i./hm2 , so it was recognized as the sensitive biotype. The biotypes collected from Caowei Town of Yuanjiang County （ HN3 and HN4 from direct seeding rice fields） and Zhongyukou Town of Nanxian County presented very high resistance to quinclorac ; the ECho values were 1 999. 122 5, 1 664.544 6 and 1 022.739 5 g a. i./hm2, and their resistance indexes reached 28.46, 23.69 and 14.56, respectively, indicating that these throe biotypes hod high level of resistance to quinclo- rac. EC50 values of about 22 biotypes were ranged from 144. 837 3 to 408.289 4 g a. i./hm2, and resistance indexes were ranged from 2.06 to 5.81, indicating that these biotypes hod produced resistance to quinclorac. EC50 values of other 10 biotypes were ranged from 98.032 6 to 138. 660 9 g a. i./hm2, and the resistance in- dexes were ranged from 1.40 to 1.97, indicating that the sensitivity of barnyard grass in these places was decreasing. [ Conclusion] The paper provided guidance for scientific and rational use of quinclorac in paddy fields.
基金National Key R&D Program of China(Nos.2019YFD1001800 and 2017YFD0201604)the National Natural Science Foundation of China(Nos.31772125 and 31972234)。
文摘Pepino mosaic virus(PepMV)causes severe disease in tomato and other Solanaceous crops around globe.To effectively study and manage this viral disease,researchers need new,sensitive,and high-throughput approaches for viral detection.In this study,we purified PepMV particles from the infected Nicotiana benthamiana plants and used virions to immunize BALB/c mice to prepare hybridomas secreting anti-PepMV monoclonal antibodies(mAbs).A panel of highly specific and sensitive murine mAbs(15B2,8H6,23D11,20D9,3A6,and 8E3)could be produced through cell fusion,antibody selection,and cell cloning.Using the mAbs as the detection antibodies,we established double antibody sandwich enzyme-linked immunosorbent assay(DAS-ELISA),Dot-ELISA,and Tissue print-ELISA for detecting PepMV infection in tomato plants.Resulting data on sensitivity analysis assays showed that both DAS-ELISA and Dot-ELISA can efficiently monitor the virus in PepMV-infected tissue crude extracts when diluted at 1:1310720 and 1:20480(weight/volume ratio(w/v),g/mL),respectively.Among the three methods developed,the Tissue print-ELISA was found to be the most practical detection technique.Survey results from field samples by the established serological approaches were verified by reverse transcription polymerase chain reaction(RT-PCR)and DNA sequencing,dem on strati ng all three serological methods are reliable and effective for monitoring PepMV.An ti-PepMV mAbs and the newly developed DAS-ELISA,Dot-ELISA,and Tissue print-ELISA can benefit PepMV detection and field epidemiological study,and management of this viral disease,which is already widespread in tomato plants in Yunnan Province of China.
基金supported by the Knowledge Innovation Program of the Chinese Academy of Sciences(KSCX2-EW-N-01)
文摘One sequence tagged site marker Subl-1 and twice submergence stress method were used in selection of submergence tolerant homozygous line from Sub-lBS, a submergence tolerant, bentazon sensitive and photoperiod-sensitive and/or thermo-sensitive genic male sterile line that developed by our laboratory. The results revealed that the original Sub-lBS was heterozygous in SublA-1 locus even though it was identical in almost all of agronomical traits and the segregation of SublA-1 was in accordance with Mendelian law based on chi-square test. And then the original Sub-IBS was divided into two groups： one was ofSublA-1 introgression and the other was not; and the two groups were tested by twice submergence stress method. After the first submergence stress that lasted for 12 d, the average plant heights were significant difference at the 1% level between the two groups. After recovery for 10 d, the second submergence stress sustained for 18 d was carried on; and the group with SublA-1 gene was found apparently tolerant than the other group in submergence tolerance.
基金supported by the Special Fund for Agro-Scientific Research in the Public Interest, China (201303028)the National Natural Science Foundation of China (31571977)
文摘Soybean chlorotic mottle virus(SbCMV)was first detected from soybean plants in Jiangxi Province of China by high throughput sequencing and was confirmed by PCR.The complete nucleotide sequence of NC113 was determined to be 8210 nucleotides,and shared the highest similarity(91.7%)with sequences of SbCMV that was only reported in Japan.It encodes nine putative open reading frames(ORFs Ia,Ib and Ⅱ-Ⅷ),and contains a large intergenic region located at nucleotide 5976-6512 between ORFs VI and VII.Sequence analysis and phylogenetic tree indicated that NC113 is an isolate of SbCMV,and is more related to the soymoviruses Blueberry red ringspot virus(BRRSV),Peanut chlorotic streak virus(PCSV)and Cestrum yellow leaf curling virus(CmYLCV)than to other representative members in the Caulimoviridae family.Field survey of 472 legume plants from Jiangxi and Zhejiang provinces showed SbCMV was only detected from soybean in Nanchang City with a low incidence rate.This is the first report of Soybean chlorotic mottle virus identified in China.
基金supported by the State Key Science and Technology Programme on Breeding of New Genetically Modified Organisms, China (2011ZX08001-003 and 2016ZX08001-003)the Open Research Fund of State Key Laboratory of Hybrid Rice (Hunan Hybrid Rice Research Center), China
文摘In this study, the characteristic of three transformants named as B1C106-1, B1C106-2, and B1C106-3 were studied that carried three innate resistant genes Bph14, Bph15, and Xa23, and two enthetic resistant genes CrylCa# and Bar. The five resistant genes were all verified by PCR and the two enthetic genes were identified in single copy insertion by Southern blot. At tillering stage, the CrylC and PAT （phosphinothricin acetyl transferase） protein contents in leaf, sheath, and stem of T2 generation were in the similar pattern： leaf〉stem〉sheath, and showed significant difference （P〈0.01） among three organs. The average contents of CrylC protein in plant of B1C106-1, B1C106-2, and BIC106-3 were 12.95, 6.57, and 11.30 μg g-1, respectively, and showed significant difference （P〈0.01） among them. However, the average contents of PAT in plant of B1C106-1, B1C106-2, and B1C106-3 were 28.54, 27.66, and 28.02 pg g-1, respectively, and there were no significant difference among three transformants. The glufosinate tolerable concentration of three transformants of T3 generation reached at least 6 g L-1, and the mortality of rice leaf rollers were above 97.4% in 5 days after being fed with fresh transformants＇ leaves. The CrylC protein toxicity was also assessed by silkworms, and the mortality of silkworms feeding mulberry leaves smeared with Cry1C protein extracts of leaves of B1 C106-1, BIC106-2, and B1 C106-3 were 90, 67.8, and 87.8%, respectively, that were positive correlation （r=0.993） with CrylC protein contents in plant of three transformants. The three transformants also maintained high resistance to brown planthopper and bacterial blight as the original version. The above results indicate the tetra-resistant rice germplasm was well-developed by pyramiding innate and enthetic resistant genes in an elite line to provide with resistances of glufosinate, rice leaf roller, brown planthopper, and bacterial blight.
基金Supported by Changde Science and Technology Transformation and Promotion Service Project[2019][CCN][0051-000].
文摘[Objectives]A codominance functional marker of the broad-spectrum bacterial blight resistance gene,Xa7,of rice was identified for accurate detection,generation tracking,and differentiation between homozygous and hemizygous genotypes of the gene.[Methods]A potential functional marker containing four primers was designed using Premier 5 software and based on the differences on the sequences of Xa7,xa7,and allele-free genomes.The molecular distinctness of the marker in different materials was verified by PCR.Three crossbreed lines of Xa7 and their parents were inoculated with seven bacterial blight strains at the booting stage to examine the affected agronomic traits at maturation.[Results]The homozygous R084 of Xa7 could be amplified into a 91 bp band and the Nip free of allele with a 153 bp band,while the heterozygote Nip/R084,91 bp and 153 bp bands.The candidate codominance marker,Xa7fun,amplified fragments that matched the predicted target bands.No 91 bp fragment was amplified from 18 germplasms of varied types,indicating a lack of Xa7 in them.Whereas Ry1,Ry2 and Ry3 had a 91 bp band,suggesting the inclusion of homozygous Xa7.Under an elevated temperature,Huazhan responded to the seven bacterial blight pathogens as highly susceptible(HS),intermediate susceptible(MS),or susceptible(S);R084 to six of the seven pathogens(HNA1-4,FuJ,GDA2,GD1358,PX086,and YN24)as highly resistant(HR),intermediate resistant(MR)or resistant(R);Ry-1 to five pathogens(GDA2,HNA1-4,FuJ,GD1358,and YN24)as HR or MR;Ry-2 to five pathogens(GDA2,GD1358,HNA1-4,PXO86,and YN24)as HR or R;and Ry-3 to 6 pathogens(HNA1-4,FuJ,GDA2,GD1358,PXO86,and YN24)as HR or MR.Therefore,the infiltration of Xa7 in the improved crossbred lines RY-1,RY-2,and RY-3 significantly accentuated the blight resistance of Huazhan.[Conclusions]Homozygous or hemizygous Xa7 could be accurately differentiated by the currently identified codominance functional marker Xa7 fun.The Xa7 introgression did not significantly alter the critical agronomic traits in the hybridization from generation to generation and could be safely applied in breeding rice varieties with bacterial blight resistance.
