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
This paper reports the findings regarding morphology, DNA markers and fruit quality of Solanum muricatum L., a newly introduced fruit in Hazara University, Pakistan, and other locally grown members of the Solanaceae f...This paper reports the findings regarding morphology, DNA markers and fruit quality of Solanum muricatum L., a newly introduced fruit in Hazara University, Pakistan, and other locally grown members of the Solanaceae family. In this study, RAPD (Random Amplified Polymorphic NA) based PCR report on phylogenetic affinities of pepino with brinjal, chilies, potato and tomato was also included. Total genomic DNA was isolated and 120 arbitrary decamers were applied to the genomic DNA of all these species. Only five out of the 120 decamers produced 27 polymorphic loci with the range of 400 - 1400 bp. We observed the specificity of markers, and primer B-08 amplified 700 bp locus linked to Solanum muricatum L., (Pepino), which could be used to identify this particular species of this family. Cluster analysis was also performed using the DNAMAN software (version 5.2.2.0) against the bivariate data collected from the products of several decamers. The UPGMA (Unweighted pair Group Method with Arithmetic Mean) analyses depicted three distinct groups, i.e. group I sorted out brinjal-tomato, group II sorted out chili-potato and pepino sorted out into its independent group III. Though on the basis of morphological traits pepino clustered with tomato, its DNA analyses proved it as a distinct species, far more related to the tomato. Moreover a total six parameters in biochemical analysis were studied, which revealed that pepino is a juicy fruit with maximum 93.25% moisture. This study could be helpful to maximize the new crop pepino in Pakistan.展开更多
The some physical and chemical properties values ofpepino fruit were determined. The mineral elements （AI, B, Ca, Cu, Fe, K, Mg, Mn, Mo, Na, Ni, P, Se and Zn） of fruit were analysed by Inductively Coupled Plasma Ato...The some physical and chemical properties values ofpepino fruit were determined. The mineral elements （AI, B, Ca, Cu, Fe, K, Mg, Mn, Mo, Na, Ni, P, Se and Zn） of fruit were analysed by Inductively Coupled Plasma Atomic Emission Spectrometry （ICP-AES）. Organic acid contents were analysed via HPLC equipped Ca （3,256.96 mg/kg）, K （43,465.60 mg/kg）, P （7,907.32 mg/kg） Na （1,496.13 mg/kg） and Mg （2,541.12 mg/kg） were established as major minerals in fruits. These results show that pepino fruit may be useful for the evaluation of dietary information in important food crops.展开更多
Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s co...Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s countries. Prevention, based on early virus detection is the only effective control measure. Monoclonal antibodies appeared to be very useful tool. The authors used for the production of monoclonal antibodies hybridomas technique, by fusing spleen cells of immunized BALB/c mice to PepMV and SP2/O cancerous cells. The aim of this work is to produce hybridomas producers of Mab that could be used for ELISA in Morocco. At the same time, these efforts will serve to decrease expenses of producers concerning phytosanitory control. We obtained 16 hybridomas lines producers of Mab specific for PepMV. They were tested for efficiencies in ELISA and two lines were retained for production of Mab on large scale （1B 11-G 10 and 5A l-G5）. Isotyping of these two lines showed that they are belonging to IgG 1 class and easily purified by affinity chromatography in agarose column by protein A. The conjugation of these two antibodies to alkaline phosphatase has been verified by DAS-ELISA. These antibodies will enable to diagnose the disease from infected tomato plants, integrating several serological tests to control it and target the actions of struggles.展开更多
[Objectives]This study was conducted to explore the feasibility of planting pepino(Solanum muricatum Ait.)in Qinghai Province,and to understand the cultivation quality and yield in various areas of Qinghai.[Methods]Wi...[Objectives]This study was conducted to explore the feasibility of planting pepino(Solanum muricatum Ait.)in Qinghai Province,and to understand the cultivation quality and yield in various areas of Qinghai.[Methods]With small fruit type of pepino as the material,four areas in eastern Qinghai were selected to determine the agronomic traits,yield and quality indexes of pepino.[Results]Under the same cultivation conditions,there were some differences in the cultivation status of pepino,but overall,pepino fruit had higher quality.Various physiological indexes were correlated with quality and yield.[Conclusions]This study clarified the specific cultivation situation of pepino in Qinghai Province,and evaluated the quality and yield of pepino,providing strong data support for the promotion and planting of pepino in various regions in the future.展开更多
Autophagy plays an active anti-viral role in plants.Increasing evidence suggests that viruses can inhibit or manipulate autophagy,thereby winning the arms race between plants and viruses.Here,we demonstrate that overe...Autophagy plays an active anti-viral role in plants.Increasing evidence suggests that viruses can inhibit or manipulate autophagy,thereby winning the arms race between plants and viruses.Here,we demonstrate that overexpression of an m^(6)A writer from Solanum lycopersicum,SlHAKAI,could negatively regulate pepino mosaic virus(PepMV)infection,inhibit viral RNA and protein accumulations by affecting viral m^(6)A levels in tomato plants and vice versa.The PepMV-encoded RNA-dependent RNA polymerase(RdRP)directly interacts with SlHAKAI and reduces its protein accumulation.The RdRP-mediated decreased protein accumulation of SlHAKAI is sensitive to the autophagy inhibitor 3-methyladenine and is compromised by knocking down a core autophagy gene.Furthermore,PepMV RdRP could interact with an essential autophagy-related protein,SlBeclin1.RdRP,SlHAKAI,and SlBeclin1 interaction complexes form bright granules in the cytoplasm.Silencing of Beclin1 in Nicotiana benthamiana plants abolishes the RdRP-mediated degradation of SlHAKAI,indicating the requirement of Beclin1 in this process.This study uncovers that the PepMV RdRP exploits the autophagy pathway by interacting with SlBeclin1 to promote the autophagic degradation of the SlHAKAI protein,thereby inhibiting the m^(6)A modification-mediated plant defense responses.展开更多
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.
文摘This paper reports the findings regarding morphology, DNA markers and fruit quality of Solanum muricatum L., a newly introduced fruit in Hazara University, Pakistan, and other locally grown members of the Solanaceae family. In this study, RAPD (Random Amplified Polymorphic NA) based PCR report on phylogenetic affinities of pepino with brinjal, chilies, potato and tomato was also included. Total genomic DNA was isolated and 120 arbitrary decamers were applied to the genomic DNA of all these species. Only five out of the 120 decamers produced 27 polymorphic loci with the range of 400 - 1400 bp. We observed the specificity of markers, and primer B-08 amplified 700 bp locus linked to Solanum muricatum L., (Pepino), which could be used to identify this particular species of this family. Cluster analysis was also performed using the DNAMAN software (version 5.2.2.0) against the bivariate data collected from the products of several decamers. The UPGMA (Unweighted pair Group Method with Arithmetic Mean) analyses depicted three distinct groups, i.e. group I sorted out brinjal-tomato, group II sorted out chili-potato and pepino sorted out into its independent group III. Though on the basis of morphological traits pepino clustered with tomato, its DNA analyses proved it as a distinct species, far more related to the tomato. Moreover a total six parameters in biochemical analysis were studied, which revealed that pepino is a juicy fruit with maximum 93.25% moisture. This study could be helpful to maximize the new crop pepino in Pakistan.
文摘The some physical and chemical properties values ofpepino fruit were determined. The mineral elements （AI, B, Ca, Cu, Fe, K, Mg, Mn, Mo, Na, Ni, P, Se and Zn） of fruit were analysed by Inductively Coupled Plasma Atomic Emission Spectrometry （ICP-AES）. Organic acid contents were analysed via HPLC equipped Ca （3,256.96 mg/kg）, K （43,465.60 mg/kg）, P （7,907.32 mg/kg） Na （1,496.13 mg/kg） and Mg （2,541.12 mg/kg） were established as major minerals in fruits. These results show that pepino fruit may be useful for the evaluation of dietary information in important food crops.
文摘Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s countries. Prevention, based on early virus detection is the only effective control measure. Monoclonal antibodies appeared to be very useful tool. The authors used for the production of monoclonal antibodies hybridomas technique, by fusing spleen cells of immunized BALB/c mice to PepMV and SP2/O cancerous cells. The aim of this work is to produce hybridomas producers of Mab that could be used for ELISA in Morocco. At the same time, these efforts will serve to decrease expenses of producers concerning phytosanitory control. We obtained 16 hybridomas lines producers of Mab specific for PepMV. They were tested for efficiencies in ELISA and two lines were retained for production of Mab on large scale （1B 11-G 10 and 5A l-G5）. Isotyping of these two lines showed that they are belonging to IgG 1 class and easily purified by affinity chromatography in agarose column by protein A. The conjugation of these two antibodies to alkaline phosphatase has been verified by DAS-ELISA. These antibodies will enable to diagnose the disease from infected tomato plants, integrating several serological tests to control it and target the actions of struggles.
基金Supported by Science and Technology Demonstration of Rural Revitalization in Jianzha County(2023-NK-X01)。
文摘[Objectives]This study was conducted to explore the feasibility of planting pepino(Solanum muricatum Ait.)in Qinghai Province,and to understand the cultivation quality and yield in various areas of Qinghai.[Methods]With small fruit type of pepino as the material,four areas in eastern Qinghai were selected to determine the agronomic traits,yield and quality indexes of pepino.[Results]Under the same cultivation conditions,there were some differences in the cultivation status of pepino,but overall,pepino fruit had higher quality.Various physiological indexes were correlated with quality and yield.[Conclusions]This study clarified the specific cultivation situation of pepino in Qinghai Province,and evaluated the quality and yield of pepino,providing strong data support for the promotion and planting of pepino in various regions in the future.
基金This work was funded by the National Key Research and Development Program of China(2021YFD1400400)to FLthe National Natural Science Foundation of China(31930089 and 31972244)to XZ and FL.
文摘Autophagy plays an active anti-viral role in plants.Increasing evidence suggests that viruses can inhibit or manipulate autophagy,thereby winning the arms race between plants and viruses.Here,we demonstrate that overexpression of an m^(6)A writer from Solanum lycopersicum,SlHAKAI,could negatively regulate pepino mosaic virus(PepMV)infection,inhibit viral RNA and protein accumulations by affecting viral m^(6)A levels in tomato plants and vice versa.The PepMV-encoded RNA-dependent RNA polymerase(RdRP)directly interacts with SlHAKAI and reduces its protein accumulation.The RdRP-mediated decreased protein accumulation of SlHAKAI is sensitive to the autophagy inhibitor 3-methyladenine and is compromised by knocking down a core autophagy gene.Furthermore,PepMV RdRP could interact with an essential autophagy-related protein,SlBeclin1.RdRP,SlHAKAI,and SlBeclin1 interaction complexes form bright granules in the cytoplasm.Silencing of Beclin1 in Nicotiana benthamiana plants abolishes the RdRP-mediated degradation of SlHAKAI,indicating the requirement of Beclin1 in this process.This study uncovers that the PepMV RdRP exploits the autophagy pathway by interacting with SlBeclin1 to promote the autophagic degradation of the SlHAKAI protein,thereby inhibiting the m^(6)A modification-mediated plant defense responses.
