Parasites,such as flatworms,nematodes,and pentastomes,are commonly found inhabiting reptiles.In this study,we examined parasite infection status in 10 squamate species,including 5 snake species and 5 lizard species.We...Parasites,such as flatworms,nematodes,and pentastomes,are commonly found inhabiting reptiles.In this study,we examined parasite infection status in 10 squamate species,including 5 snake species and 5 lizard species.We identified one genuslevel and nine species-level parasites in these hosts,which belong to the phyla Platyhelminthes,Nemathelminthes,and Arthropoda.The overall infection rate was 45%,with significant differences in infection rates between lizard and snake species.Parasites in snakes included the Kalicephalus sp.,Ophidascaris filaria,Ophiotaenia bungari,Raillietiella orientalis and Spirometra erinaceieuropaei,and parasitic parasites in lizards were mainly composed of Meteterakis japonica,Plagiorchis koreanus,Spauligodon carbonelli,Spauligodon saxicolae and Strongyluris calotis,respectively.There were significant differences in infection rates among different species and regions,and even among different tissues of the same species.The overall investigation of squamate parasites in China will provide some basic information for public health safety.展开更多
[Objective] This study aimed to investigate the prevalence and variation of porcine kobuvirus （PKV） in suckling piglets in China. [Method] In 2013-2014, 224 feces samples from suckling piglets with diarrhea in 27 pi...[Objective] This study aimed to investigate the prevalence and variation of porcine kobuvirus （PKV） in suckling piglets in China. [Method] In 2013-2014, 224 feces samples from suckling piglets with diarrhea in 27 pig farms of five provinces in China were collected to detect 3D genes of PKV with RT-PCR method; the sequences and genetic variation of 29 PKV 3D genes were analyzed. [Result] Total positive rate of PKV in feces samples from suckling piglets with diarrhea was 65.18% （146/224）; total positive rate of PKV in pig farms was 85,2% （23/27）; nucleotide sequences and the deduced amino acid sequences of 29 PKV 3D genes shared 87.0%-100% and 92.7%-100% homologies with six PKV-related 3D sequences, respectively. [Conclusion] PKV infection is prevalent in suckling piglets in China; PKV 3D genes exhibit high diversity.展开更多
We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticillium albo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer （...We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticillium albo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer （ITS） sequences of Verticillium spp., a pair of species-specific primers, Vaal/Vaa2, was synthesized. After screening 17 isolates of V. alboatrum, 121 isolates from the Ascomycota, B asidiomycota, Deuteromycota, and Oomycota, the Vaal/Vaa2 primers amplified only a single PCR band of approximately 330 bp from V. albo-atrum. The detection sensitivity with primers Vaal/Vaa2 was 10 fg of genomic DNA. Using ITS1/ITS4 as the first-round primers, combined with Vaa1/Vaa2, the nested PCR procedures were developed, and the detection sensitivity increased 1 000-fold to 10 ag. The detection sensitivity for the soil pathogens was 100-conidiag^-1 soil. The PCR-based methods developed here could simplify both plant disease diagnosis and pathogen monitoring as well as guide plant disease management.展开更多
The ultimate goal of single-cell analyses is to obtain the biomolecular content for each cell in unicellular and multicellular organisms at different points of their life cycle under variable environmental conditions....The ultimate goal of single-cell analyses is to obtain the biomolecular content for each cell in unicellular and multicellular organisms at different points of their life cycle under variable environmental conditions.These require an assessment of:a)the total number of cells,b)the total number of cell types,and c)the complete and quantitative single molecular detection and identification for all classes of biopolymers,and organic and inorganic compounds,in each individual cell.For proteins,glycans,lipids,and metabolites,whose sequences cannot be amplified by copying as in the case of nucleic acids,the detection limit by mass spectrometry is about 105 molecules.Therefore,proteomic,glycomic,lipidomic,and metabolomic analyses do not yet permit the assembly of the complete single-cell omes.The construction of novel nanoelectrophoretic arrays and nano in microarrays on a single 1-cm-diameter chip has shown proof of concept for a high throughput platform for parallel processing of thousands of individual cells.Combined with dynamic secondary ion mass spectrometry,with 3D scanning capability and lateral resolution of 50 nm,the sensitivity of single molecular quantification and identification for all classes of biomolecules could be reached.Further development and routine application of such technological and instrumentation solution would allow assembly of complete omes with a quantitative assessment of structural and functional cellular diversity at the molecular level.展开更多
The immature embryos of wheat cultivars Liaochun10, Tiechun1 and Fengqiang3 were bombarded with gold particles coated with pti5 vp16 by gene gun and disease resistant regenerated plants were attained. In order to...The immature embryos of wheat cultivars Liaochun10, Tiechun1 and Fengqiang3 were bombarded with gold particles coated with pti5 vp16 by gene gun and disease resistant regenerated plants were attained. In order to confirm that the plants are genuine transformed ones, a series of molecular tests were conducted as follows. Firstly, transient GUS expression test on embryos two days after bombardment was done. There were many obvious blue spots produced on the surface of bombarded embryos after GUS staining, in which the maximum reached to 85 blue spots per embryo. Secondly, PCR test was performed with DNA from the regenerated plants obtained after double selection with ppt. 6 plants were found PCR test positive. At last, further verification analysis using dot hybridization and southern blotting was carried out on those PCR positive plants and the strong hybridization signals appeared as expected. All the above tests were uniformly indicated that the disease resistant regenerated plants were true transgenic plants. When inoculated with Blumeria graminis, transgenic wheat plants of PCR positive results were mostly resistant(R) after 7 days, and resis tant, moderate resistant(MR), moderate susceptible(MS) at 14 days respectively. The disease severity of them was distinctively lighter than that of control.展开更多
According to the quarantine procedures for sugarcane introduction, thirty-three sugarcane varieties/materials from the United States, Bangladesh, Thailand and some other countries were quarantined. The result showed t...According to the quarantine procedures for sugarcane introduction, thirty-three sugarcane varieties/materials from the United States, Bangladesh, Thailand and some other countries were quarantined. The result showed that main diseases and pests that are concerned in quarantine were undiscovered. In addition, molecular detection results of Fiji disease and leaf scald disease were negative ; detection results of mosaic in six varleties/materials were positive; detection results of yellow leaf syndrome in eight varieties/materials were positive.展开更多
The present PCR assay was conducted to develop rapid and sensitive detection of Phytophthora colocasiae,in order to provide a robust and reliable tool for healthy seedling production of taro and limiting the transmiss...The present PCR assay was conducted to develop rapid and sensitive detection of Phytophthora colocasiae,in order to provide a robust and reliable tool for healthy seedling production of taro and limiting the transmission and spread of the causal organism of taro leaf blight in taro planting regions.The samples were used to extract total DNA and to be detected by PCR with P.colocasiae specific primer pairs PCSP-RL F/PCSP-RL R and PCSP-T F/PCSP-T R,respectively.Distinct fragments of about 200 bp and 240 bp were amplified by PCR using primers PCSP-RL F/PCSP-RL R and PCSP-T F/PCSP-T R,respectively.The analysis of the nucleotide sequence of the PCR products were found to be 99% identical to sequence of RAS-related protein (Ypt1) and phospho-ribosylanthranilate isomerase (TRP1) in P.colocasiae,respectively.It is concluded that rapid and sensitive developed PCR assay for detection of P.colocasiae could be used in routine diagnosis and aid in management practices to mitigate taro leaf blight.展开更多
Cystic echinococcosis (CE) is a prevalent zoonotic disease caused by Echinococcus granulosus, with a cosmopolitan distribution. The parasite is transmitted cyclically between canines and numerous intermediate herbivor...Cystic echinococcosis (CE) is a prevalent zoonotic disease caused by Echinococcus granulosus, with a cosmopolitan distribution. The parasite is transmitted cyclically between canines and numerous intermediate herbivorous livestock animals. Also, other Taeniid tapeworms could infect domestic dogs and they pose significant veterinary and public health concerns worldwide. This study aimed to develop a sensitive molecular method for detecting Echinococcus spp. DNA in dog fecal samples using next-generation sequencing (NGS). A set of PCR primers targeting conserved regions of Taeniid tapeworms’ 18s rRNA genes was designed and tested for amplifying genomic DNA from various tapeworm species. The PCR system demonstrated high sensitivity, amplifying DNA from all tested tapeworm species, with differences observed in amplified band sizes. The primers were adapted for NGS analysis by adding forward and reverse adapters, enabling the sequencing of amplified DNA fragments. Application of the developed PCR system to dog fecal samples collected from Yatta town, Palestine, revealed the presence of E. granulosus DNA in five out of 50 samples. NGS analysis confirmed the specificity of the amplified DNA fragments, showing 98% - 99% similarity with the 18s rDNA gene of E. granulosus. This study demonstrates the utility of NGS-based molecular methods for accurate and sensitive detection of Echinococcus spp. in dog fecal samples, providing valuable insights for epidemiological surveillance and control programs of echinococcosis in endemic regions.展开更多
Surface plasmonic resonance(SPR)has been a corner stone for approaching single molecular detection due to its highsensitivity capability and simple detection mechanism,and has brought major advancements in biomedicine...Surface plasmonic resonance(SPR)has been a corner stone for approaching single molecular detection due to its highsensitivity capability and simple detection mechanism,and has brought major advancements in biomedicine and life science technology.Over decades,the successful integration of SPR with versatile techniques has been demonstrated.However,several crucial limitations have hindered this technique for practical applications,such as long detection time and low overall sensitivity.This review aims to provide a comprehensive summary of existing approaches in enhancing the performance of SPR sensors based on“passive”and“active”methods.Firstly,passive enhancement is discussed from a material aspect,including signal amplification tags and modifications of conventional substrates.Then,the focus is on the most popular active enhancement methods including electrokinetic,optical,magnetic,and acoustic manipulations that are summarized with highlights on their advantageous features and ability to concentrate target molecules at the detection sites.Lastly,prospects and future development directions for developing SPR sensing towards a more practical,single molecular detection technique in the next generation are discussed.This review hopes to inspire researchers’interests in developing SPR-related technology with more innovative and influential ideas.展开更多
It is necessary to rapidly diagnose diseases, identify and monitor the pathogens to achieve scientific and effective control of major diseases in sugarcane. In the present study, the molecular techniques for rapid det...It is necessary to rapidly diagnose diseases, identify and monitor the pathogens to achieve scientific and effective control of major diseases in sugarcane. In the present study, the molecular techniques for rapid detection of 13 pathogens that cause 10 important diseases of sugarcane including smut, rust, leaf scald, ratoon stunting, red stripe, mosaic, Fiji, yellow leaf, white leaf and bacilliform virus were established by Sugarcane Research Institute, Yunnan Academy of Agricul- tural Sciences after years of effort. The results will provide scientific basis for effective diagnosis and control of sugarcane diseases, detection of virus-free seedlings and quarantine management of exotic species.展开更多
Citrus tristeza virus （CTV） exists in citrus as a large number of distinct strains differing in biological characters. The control strategies such as mild strains cross protection （MSCP） require a clear understand...Citrus tristeza virus （CTV） exists in citrus as a large number of distinct strains differing in biological characters. The control strategies such as mild strains cross protection （MSCP） require a clear understanding of the characterization of CTV. For better understanding of the structure of CTV population and the relationship between molecular and biological characterization, 72 CTV samples collected from five provinces in China were studied, using biological indexing, p25/Hinf I restriction fragment length polymorphism （RFLP）, multiple molecular markers, and bidirectional RT-PCR assay. The mixture of severe stem pitting isolates was found to be dominant in the field. CTV isolates with p25/Hinf Ⅰ RFLP group 3 and p23/BD-PCR group Ⅰ, Ⅲ were the main cause of epidemics, and most CTV isolates were found to be the mixture of T30 and VT genotypes. More accurate identification of strain mixtures in the field and better understanding of the biological traits of the isolates may be achieved by applying the three molecular detection methods simultaneously.展开更多
Worldwide there has been a significant increase in the incidence of oropharyngeal squamous cell carcinoma(OPSCC)etiologically attributed to oncogenic human papillomavirus(HPV).Reliable and accurate identification and ...Worldwide there has been a significant increase in the incidence of oropharyngeal squamous cell carcinoma(OPSCC)etiologically attributed to oncogenic human papillomavirus(HPV).Reliable and accurate identification and detection tools are important as the incidence of HPV-related cancer is on the rise.Several HPV detection methods for OPSCC have been developed and each has its own advantages and disadvantages in regard to sensitivity,specificity,and technical difficulty.This review summarizes our current knowledge of molecular methods for detecting HPV in OPSCC,including HPV DNA/RNA polymerase chain reaction(PCR),loop-mediated isothermal amplification(LAMP),p16 immunohistochemistry(IHC),and DNA/RNA in situ hybridization(ISH)assays.This summary may facilitate the selection of a suitable method for detecting HPV infection,and therefore may help in the early diagnosis of HPV-related carcinoma to reduce its mortality,incidence,and morbidity.展开更多
Fusarium oxysporum f. sp. conglutinans （Foc） is the causal agent of Fusarium wilt disease of Brassica oleracea. A rapid, accurate, and reliable method to detect and identify plant pathogens is vitally important to i...Fusarium oxysporum f. sp. conglutinans （Foc） is the causal agent of Fusarium wilt disease of Brassica oleracea. A rapid, accurate, and reliable method to detect and identify plant pathogens is vitally important to integrated disease management. In this study, using a comparative genome analysis among Fusarium oxysporum （Fo）, we developed a Foc-specific primer set （Focs-l/Focs-2） and established a multiplex-PCR assay. In the assay, the Focs-1/Focs-2 and universal primers for Fusarium species （W106PJF106S） could be used to detect Foc isolates in a single PCR reaction. With the optimized PCR parameters, the multiplex-PCR assay showed a high specificity for detecting Foc and was very sensitive to detect as little as 100 pg of pure Foc genomic DNAor 1 000 spores in 1 g of twice-autoclaved soil. We also demonstrated that Foc isolates were easily detected from infected plant tissues, as well as from natural field soils, using the multiplex-PCR assay. To our knowledge, this is a first report on detection Fo by comparative genomic method.展开更多
In China, the purity of maize hybrid strain is discomforting to the development of seed industrialization. Finding a new method for reproduction of maize hybrid strain is necessary. In this study, using particle bomba...In China, the purity of maize hybrid strain is discomforting to the development of seed industrialization. Finding a new method for reproduction of maize hybrid strain is necessary. In this study, using particle bombardment, barstar gene was transferred into maize inbred line 18-599 （White）, which is an antiviral and high quality maize inbred line. By molecular detection of the anther of transgenic maize, two plants transferred with barstar gene were gained in this study, which are two restorer lines. The two plants showed normal male spike, and lively microspores. But the capacity of the two restorer lines should be studied in the future. The aim of this study is to find a new method of reproduction of maize hybrid strain using engineering restorer lines and engineering sterility lines by gene engineering technology.展开更多
Lymnaeid snails are key intermediate hosts for the development and survival of Fasciola spp.,the causative agent of Fascioliasis which are economically important parasites infecting humans and livestock globally.The c...Lymnaeid snails are key intermediate hosts for the development and survival of Fasciola spp.,the causative agent of Fascioliasis which are economically important parasites infecting humans and livestock globally.The current control method for treating Fascioliasis is heavily reliant on anthelmintic drugs,particularly Triclabendazole(TCBZ)which has resulted in drug-resistant parasites and poses significant risk as there are no long-term efficacious alternatives available.Sustainable control measures at the farm level could include both parasite and snail control will play an important role in Fasciola spp.control and reduce the reliance on anthelmintic drugs.Implementation of such sustainable control measures requires effective identification of snails on the property however Lymnaeid snails are small and difficult to physically locate.Snail identification using an environmental DNA approach is a recent approach in which physically locating snails are not required.Austropeplea tomentosa,is the primary intermediate snail host for F.hepatica transmission in South-East Australia and we present an in-field loop-mediated isothermal amplification and water filtering method for the detection of A.tomentosa eDNA from water samples to improve current surveillance methods.This methodology is highly sensitive with a detection limit of 5×10^(−6)ng/μL,detected in<20 minutes,with cumulative sample preparation and amplification time under 1 hour.This proposed workflow could assist in monitoring areas to determine the risk of Fascioliasis infection and implement strategies to manage snail populations to ultimately reduce the risk of infection for humans and livestock.展开更多
To rapidly obtain high-quality genomic DNA from Chenopodium quinoa Willd, the genomic DAN in different tissues （leaves, stems and roots） of Chenopodi- um quinoa Willd was extracted by modified CTAB method, SDS metho...To rapidly obtain high-quality genomic DNA from Chenopodium quinoa Willd, the genomic DAN in different tissues （leaves, stems and roots） of Chenopodi- um quinoa Willd was extracted by modified CTAB method, SDS method and high- salt Iow-pH method, respectively. The quality and yield of extracted DNA was deter- mined using agarose gel electrophoresis and UV spectrophotometry. At the same time, the PCR-SSR and SSCP molecular detection was also performed. The results showed that the gel test strips, without obvious decomposition, of all the extraction methods were relatively obvious; the genomic DNA yield extracted by modified CTAB method was highest, followed by that by SDS method, and the genomic DNA extracted by high-salt Iow-pH method was lowest： the genomic DNA yields extracted by different methods from Chenopodium quinoa Wiltd leaves were all high- er than those from roots and stems; the quality of Chenopodium quinoa Willd ge- nomic DNA extracted by modified CTAB method and high-salt Iow-pH method was better, and polyphenols, polysaccharides and other impurities were removed more completely. The PCR-SSR and SSCP detection results showed that the genomic DNA extracted by different methods from different tissues of Chenopodium quinoa Willd all could be better amplified, and high-quality strips could be obtained. So the Chenopodium quinoa Willd genomic DNA extracted by the three methods all can be used for subsequent molecular biology research.展开更多
[Objective] The aim of this study was to observe the agronomic,yield and quality traits of Ningchun 4 and its parents under the same environmental conditions,as well as to carry out molecular detection on the backgrou...[Objective] The aim of this study was to observe the agronomic,yield and quality traits of Ningchun 4 and its parents under the same environmental conditions,as well as to carry out molecular detection on the background genes distribution of the corresponding traits.[Method]Ningchun 4 and its parents Sonora 64,Hongtu,Abbondanza,Quality were used as materials to detect the agronomic and quality traits,as well as to analyze the genetic variation laws by molecular determination method.[Result]Ningchun 4 had inherited the advantages of bigger spike,red and hard grain from Sonora 64 and higher 1 000-grain weight from Hongtu.However,it had also inherited the disadvantages of late-maturing from Sonora 64 and lower tillering ability from Hongtu;the grain quality of Ningchun 4 was slightly lower than the Sonora 64;Ningchun 4 had high quality subunit of 5＋10,which had good dough rheological properties.[Conclusion]Ningchun 4 had inherited the long photoperiod characteristics and no-resistant to slow-leaf rusting and stripe rust characteristics from Hongtu and low PPO activity,high yellow pigments content from parents.展开更多
White spot syndrome virus (WSSV),Taura syndrome virus (TSV) and Infectious hypodermal and haematopoietic necrosis virus (IHHNV) are three shrimp viruses responsible for major pandemics affecting the shrimp farming ind...White spot syndrome virus (WSSV),Taura syndrome virus (TSV) and Infectious hypodermal and haematopoietic necrosis virus (IHHNV) are three shrimp viruses responsible for major pandemics affecting the shrimp farming industry. Shrimps samples were collected from 12 farms in Zhejiang province,China,in 2008 and analyzed by PCR to determine the prevalence of these viruses. From the 12 sampling locations,8 farms were positive for WSSV,8 for IHHNV and 6 for both WSSV and IHHNV. An average percentage of 57.4% of shrimp individuals were infected with WSSV,while 49.2% were infected with IHHNV. A high prevalence of co-infection with WSSV and IHHNV among samples was detected from the following samples:Bingjiang (93.3%),liuao (66.7%),Jianshan (46.7%) and Xianxiang (46.7%). No samples exhibited evidence of infection with TSV in collected samples. This study provides comprehensive information of the prevalence of three shrimp viruses in Zhejiang and may be helpful for disease prevention control in this region.展开更多
The common fig（Ficus carica L.）was one of the earliest horticultural crops to be domesticated.A number of different viruses can infect fig trees including Fig mosaic virus（FMV）that has been detected in several com...The common fig（Ficus carica L.）was one of the earliest horticultural crops to be domesticated.A number of different viruses can infect fig trees including Fig mosaic virus（FMV）that has been detected in several commercial fig trees in Xinjiang,China.However,the distribution of FMV and other fig-infecting viruses in China remains unknown.In the present study,a sample from an ancient fig tree growing in Xinjiang was investigated by electron microscopy（EM）followed by PCR/RT-PCR,and FMV,Fig badnavirus 1（FBV-1）and Fig leaf mottle-associated virus 1（FLMaV-1）were detected.Fig leaf samples（252）from commercial orchards across China were subjected to PCR/RT-PCR,and FMV,FBV-1 and Fig fleck-associated virus（FFka V）were relatively abundant（44.4,48.4 and 44%,respectively）,while FLMaV-1 and Fig mild mottle-associated virus（FMMa V）were much scarcer（5.6 and 0.4%,respectively）,and FLMaV-2,Fig cryptic virus（FCV）,and Fig latent virus（FLV）were not detected.The presence of disease-causing viruses in fig trees presents a significant challenge for fig producers in China.This study may help to promote actions aimed at controlling fig viruses,especially FMV.展开更多
基金supported by grants from the National Natural Science Foundation of China (32171498 and 32171495)the Finance Science and Technology Project of Hainan Province (ZDYF2018219)the Hainan Key Program of Science and Technology (ZDXM20110008)。
文摘Parasites,such as flatworms,nematodes,and pentastomes,are commonly found inhabiting reptiles.In this study,we examined parasite infection status in 10 squamate species,including 5 snake species and 5 lizard species.We identified one genuslevel and nine species-level parasites in these hosts,which belong to the phyla Platyhelminthes,Nemathelminthes,and Arthropoda.The overall infection rate was 45%,with significant differences in infection rates between lizard and snake species.Parasites in snakes included the Kalicephalus sp.,Ophidascaris filaria,Ophiotaenia bungari,Raillietiella orientalis and Spirometra erinaceieuropaei,and parasitic parasites in lizards were mainly composed of Meteterakis japonica,Plagiorchis koreanus,Spauligodon carbonelli,Spauligodon saxicolae and Strongyluris calotis,respectively.There were significant differences in infection rates among different species and regions,and even among different tissues of the same species.The overall investigation of squamate parasites in China will provide some basic information for public health safety.
文摘[Objective] This study aimed to investigate the prevalence and variation of porcine kobuvirus （PKV） in suckling piglets in China. [Method] In 2013-2014, 224 feces samples from suckling piglets with diarrhea in 27 pig farms of five provinces in China were collected to detect 3D genes of PKV with RT-PCR method; the sequences and genetic variation of 29 PKV 3D genes were analyzed. [Result] Total positive rate of PKV in feces samples from suckling piglets with diarrhea was 65.18% （146/224）; total positive rate of PKV in pig farms was 85,2% （23/27）; nucleotide sequences and the deduced amino acid sequences of 29 PKV 3D genes shared 87.0%-100% and 92.7%-100% homologies with six PKV-related 3D sequences, respectively. [Conclusion] PKV infection is prevalent in suckling piglets in China; PKV 3D genes exhibit high diversity.
基金We thank Dr Chen Qinghe,Dr Ko Wenhong,Dr Ho Hanhin,Dr Hu Baishi,Dr Peng Jinghuo and China General Microbiological Culture Collection Center(CGMCC)for providing some isolates.This work was supported by the China National 863 Program(2003AA249020).
文摘We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticillium albo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer （ITS） sequences of Verticillium spp., a pair of species-specific primers, Vaal/Vaa2, was synthesized. After screening 17 isolates of V. alboatrum, 121 isolates from the Ascomycota, B asidiomycota, Deuteromycota, and Oomycota, the Vaal/Vaa2 primers amplified only a single PCR band of approximately 330 bp from V. albo-atrum. The detection sensitivity with primers Vaal/Vaa2 was 10 fg of genomic DNA. Using ITS1/ITS4 as the first-round primers, combined with Vaa1/Vaa2, the nested PCR procedures were developed, and the detection sensitivity increased 1 000-fold to 10 ag. The detection sensitivity for the soil pathogens was 100-conidiag^-1 soil. The PCR-based methods developed here could simplify both plant disease diagnosis and pathogen monitoring as well as guide plant disease management.
文摘The ultimate goal of single-cell analyses is to obtain the biomolecular content for each cell in unicellular and multicellular organisms at different points of their life cycle under variable environmental conditions.These require an assessment of:a)the total number of cells,b)the total number of cell types,and c)the complete and quantitative single molecular detection and identification for all classes of biopolymers,and organic and inorganic compounds,in each individual cell.For proteins,glycans,lipids,and metabolites,whose sequences cannot be amplified by copying as in the case of nucleic acids,the detection limit by mass spectrometry is about 105 molecules.Therefore,proteomic,glycomic,lipidomic,and metabolomic analyses do not yet permit the assembly of the complete single-cell omes.The construction of novel nanoelectrophoretic arrays and nano in microarrays on a single 1-cm-diameter chip has shown proof of concept for a high throughput platform for parallel processing of thousands of individual cells.Combined with dynamic secondary ion mass spectrometry,with 3D scanning capability and lateral resolution of 50 nm,the sensitivity of single molecular quantification and identification for all classes of biomolecules could be reached.Further development and routine application of such technological and instrumentation solution would allow assembly of complete omes with a quantitative assessment of structural and functional cellular diversity at the molecular level.
文摘The immature embryos of wheat cultivars Liaochun10, Tiechun1 and Fengqiang3 were bombarded with gold particles coated with pti5 vp16 by gene gun and disease resistant regenerated plants were attained. In order to confirm that the plants are genuine transformed ones, a series of molecular tests were conducted as follows. Firstly, transient GUS expression test on embryos two days after bombardment was done. There were many obvious blue spots produced on the surface of bombarded embryos after GUS staining, in which the maximum reached to 85 blue spots per embryo. Secondly, PCR test was performed with DNA from the regenerated plants obtained after double selection with ppt. 6 plants were found PCR test positive. At last, further verification analysis using dot hybridization and southern blotting was carried out on those PCR positive plants and the strong hybridization signals appeared as expected. All the above tests were uniformly indicated that the disease resistant regenerated plants were true transgenic plants. When inoculated with Blumeria graminis, transgenic wheat plants of PCR positive results were mostly resistant(R) after 7 days, and resis tant, moderate resistant(MR), moderate susceptible(MS) at 14 days respectively. The disease severity of them was distinctively lighter than that of control.
基金Supported by Special Fund for Modern Agro-industry Technology Research System(NYCYTXGXCXTD-2)Scientific Research and Technological Development Project of Guangxi Guangxi Zhuang Autonomous Region(GKN1347012-1+2 种基金GKN 1347004-2)Natural Science Foundation of Guangxi Zhuang Autonomous Region(11166-02)Internaltional Scientific Exchange Program(2013DFA31600)
文摘According to the quarantine procedures for sugarcane introduction, thirty-three sugarcane varieties/materials from the United States, Bangladesh, Thailand and some other countries were quarantined. The result showed that main diseases and pests that are concerned in quarantine were undiscovered. In addition, molecular detection results of Fiji disease and leaf scald disease were negative ; detection results of mosaic in six varleties/materials were positive; detection results of yellow leaf syndrome in eight varieties/materials were positive.
基金Sponsored by Science and Technology Major Project of Guangxi(AA17204026)Basic Research Special Fund,and Scientific and Technological Fund of Guangxi Academy of Agricultural Sciences(2016YM20,2018JZ37)Guangxi Natural Science Fund(2016GXNSFAA380195)
文摘The present PCR assay was conducted to develop rapid and sensitive detection of Phytophthora colocasiae,in order to provide a robust and reliable tool for healthy seedling production of taro and limiting the transmission and spread of the causal organism of taro leaf blight in taro planting regions.The samples were used to extract total DNA and to be detected by PCR with P.colocasiae specific primer pairs PCSP-RL F/PCSP-RL R and PCSP-T F/PCSP-T R,respectively.Distinct fragments of about 200 bp and 240 bp were amplified by PCR using primers PCSP-RL F/PCSP-RL R and PCSP-T F/PCSP-T R,respectively.The analysis of the nucleotide sequence of the PCR products were found to be 99% identical to sequence of RAS-related protein (Ypt1) and phospho-ribosylanthranilate isomerase (TRP1) in P.colocasiae,respectively.It is concluded that rapid and sensitive developed PCR assay for detection of P.colocasiae could be used in routine diagnosis and aid in management practices to mitigate taro leaf blight.
文摘Cystic echinococcosis (CE) is a prevalent zoonotic disease caused by Echinococcus granulosus, with a cosmopolitan distribution. The parasite is transmitted cyclically between canines and numerous intermediate herbivorous livestock animals. Also, other Taeniid tapeworms could infect domestic dogs and they pose significant veterinary and public health concerns worldwide. This study aimed to develop a sensitive molecular method for detecting Echinococcus spp. DNA in dog fecal samples using next-generation sequencing (NGS). A set of PCR primers targeting conserved regions of Taeniid tapeworms’ 18s rRNA genes was designed and tested for amplifying genomic DNA from various tapeworm species. The PCR system demonstrated high sensitivity, amplifying DNA from all tested tapeworm species, with differences observed in amplified band sizes. The primers were adapted for NGS analysis by adding forward and reverse adapters, enabling the sequencing of amplified DNA fragments. Application of the developed PCR system to dog fecal samples collected from Yatta town, Palestine, revealed the presence of E. granulosus DNA in five out of 50 samples. NGS analysis confirmed the specificity of the amplified DNA fragments, showing 98% - 99% similarity with the 18s rDNA gene of E. granulosus. This study demonstrates the utility of NGS-based molecular methods for accurate and sensitive detection of Echinococcus spp. in dog fecal samples, providing valuable insights for epidemiological surveillance and control programs of echinococcosis in endemic regions.
基金the National Natural Science Foundation of China(No.61905145)Guangdong Natural Science Foundation and Province Project(No.2021A1515011916)Shenzhen Science and Technology R&D and Innovation Foundation(No.JCYJ20200109105608771).
文摘Surface plasmonic resonance(SPR)has been a corner stone for approaching single molecular detection due to its highsensitivity capability and simple detection mechanism,and has brought major advancements in biomedicine and life science technology.Over decades,the successful integration of SPR with versatile techniques has been demonstrated.However,several crucial limitations have hindered this technique for practical applications,such as long detection time and low overall sensitivity.This review aims to provide a comprehensive summary of existing approaches in enhancing the performance of SPR sensors based on“passive”and“active”methods.Firstly,passive enhancement is discussed from a material aspect,including signal amplification tags and modifications of conventional substrates.Then,the focus is on the most popular active enhancement methods including electrokinetic,optical,magnetic,and acoustic manipulations that are summarized with highlights on their advantageous features and ability to concentrate target molecules at the detection sites.Lastly,prospects and future development directions for developing SPR sensing towards a more practical,single molecular detection technique in the next generation are discussed.This review hopes to inspire researchers’interests in developing SPR-related technology with more innovative and influential ideas.
基金Supported by Earmarked Fund for Modern Agro-industry Technology Research System of China(CARS-20-2-2)Earmarked fund for Modern Agroindustry Technology Research System of Yunnan Province
文摘It is necessary to rapidly diagnose diseases, identify and monitor the pathogens to achieve scientific and effective control of major diseases in sugarcane. In the present study, the molecular techniques for rapid detection of 13 pathogens that cause 10 important diseases of sugarcane including smut, rust, leaf scald, ratoon stunting, red stripe, mosaic, Fiji, yellow leaf, white leaf and bacilliform virus were established by Sugarcane Research Institute, Yunnan Academy of Agricul- tural Sciences after years of effort. The results will provide scientific basis for effective diagnosis and control of sugarcane diseases, detection of virus-free seedlings and quarantine management of exotic species.
基金supported by National Natural Science Foundation of China(30471205).
文摘Citrus tristeza virus （CTV） exists in citrus as a large number of distinct strains differing in biological characters. The control strategies such as mild strains cross protection （MSCP） require a clear understanding of the characterization of CTV. For better understanding of the structure of CTV population and the relationship between molecular and biological characterization, 72 CTV samples collected from five provinces in China were studied, using biological indexing, p25/Hinf I restriction fragment length polymorphism （RFLP）, multiple molecular markers, and bidirectional RT-PCR assay. The mixture of severe stem pitting isolates was found to be dominant in the field. CTV isolates with p25/Hinf Ⅰ RFLP group 3 and p23/BD-PCR group Ⅰ, Ⅲ were the main cause of epidemics, and most CTV isolates were found to be the mixture of T30 and VT genotypes. More accurate identification of strain mixtures in the field and better understanding of the biological traits of the isolates may be achieved by applying the three molecular detection methods simultaneously.
基金Project supported by the Universiti Sains Malaysia Research University Grant(1001/PPSG/8012345)。
文摘Worldwide there has been a significant increase in the incidence of oropharyngeal squamous cell carcinoma(OPSCC)etiologically attributed to oncogenic human papillomavirus(HPV).Reliable and accurate identification and detection tools are important as the incidence of HPV-related cancer is on the rise.Several HPV detection methods for OPSCC have been developed and each has its own advantages and disadvantages in regard to sensitivity,specificity,and technical difficulty.This review summarizes our current knowledge of molecular methods for detecting HPV in OPSCC,including HPV DNA/RNA polymerase chain reaction(PCR),loop-mediated isothermal amplification(LAMP),p16 immunohistochemistry(IHC),and DNA/RNA in situ hybridization(ISH)assays.This summary may facilitate the selection of a suitable method for detecting HPV infection,and therefore may help in the early diagnosis of HPV-related carcinoma to reduce its mortality,incidence,and morbidity.
基金supported in part by the Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture, P.R.Chinathe National Natural Science Foundation of China (31571962, 31272003)+3 种基金the National Key Technology R&D Program of China (2012BAD19B06)the Special Fund for Agro-Scientific Research in the Public Interest, China (200903049-04)the National Staple Vegetable Industry Technology System Construction Project, China (CARS-25-B-01)the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences
文摘Fusarium oxysporum f. sp. conglutinans （Foc） is the causal agent of Fusarium wilt disease of Brassica oleracea. A rapid, accurate, and reliable method to detect and identify plant pathogens is vitally important to integrated disease management. In this study, using a comparative genome analysis among Fusarium oxysporum （Fo）, we developed a Foc-specific primer set （Focs-l/Focs-2） and established a multiplex-PCR assay. In the assay, the Focs-1/Focs-2 and universal primers for Fusarium species （W106PJF106S） could be used to detect Foc isolates in a single PCR reaction. With the optimized PCR parameters, the multiplex-PCR assay showed a high specificity for detecting Foc and was very sensitive to detect as little as 100 pg of pure Foc genomic DNAor 1 000 spores in 1 g of twice-autoclaved soil. We also demonstrated that Foc isolates were easily detected from infected plant tissues, as well as from natural field soils, using the multiplex-PCR assay. To our knowledge, this is a first report on detection Fo by comparative genomic method.
文摘In China, the purity of maize hybrid strain is discomforting to the development of seed industrialization. Finding a new method for reproduction of maize hybrid strain is necessary. In this study, using particle bombardment, barstar gene was transferred into maize inbred line 18-599 （White）, which is an antiviral and high quality maize inbred line. By molecular detection of the anther of transgenic maize, two plants transferred with barstar gene were gained in this study, which are two restorer lines. The two plants showed normal male spike, and lively microspores. But the capacity of the two restorer lines should be studied in the future. The aim of this study is to find a new method of reproduction of maize hybrid strain using engineering restorer lines and engineering sterility lines by gene engineering technology.
基金supported by Cooperative Research Centres Project(CRCP)awarded to Geneworks and La Trobe University.L.T.is supported by an Australian Research Training Program scholarship and the Tim Healy Memorial Scholarship awarded by The Department of Primary Industries South Australia(PIRSA).
文摘Lymnaeid snails are key intermediate hosts for the development and survival of Fasciola spp.,the causative agent of Fascioliasis which are economically important parasites infecting humans and livestock globally.The current control method for treating Fascioliasis is heavily reliant on anthelmintic drugs,particularly Triclabendazole(TCBZ)which has resulted in drug-resistant parasites and poses significant risk as there are no long-term efficacious alternatives available.Sustainable control measures at the farm level could include both parasite and snail control will play an important role in Fasciola spp.control and reduce the reliance on anthelmintic drugs.Implementation of such sustainable control measures requires effective identification of snails on the property however Lymnaeid snails are small and difficult to physically locate.Snail identification using an environmental DNA approach is a recent approach in which physically locating snails are not required.Austropeplea tomentosa,is the primary intermediate snail host for F.hepatica transmission in South-East Australia and we present an in-field loop-mediated isothermal amplification and water filtering method for the detection of A.tomentosa eDNA from water samples to improve current surveillance methods.This methodology is highly sensitive with a detection limit of 5×10^(−6)ng/μL,detected in<20 minutes,with cumulative sample preparation and amplification time under 1 hour.This proposed workflow could assist in monitoring areas to determine the risk of Fascioliasis infection and implement strategies to manage snail populations to ultimately reduce the risk of infection for humans and livestock.
基金Supported by National Natural Science Foundation of China(31301372)Key Project of Science and Technology Plan of Zhejiang Province(2011C12030)Innovation Training Project of Zhejiang Agriculture and Forestry University(201301004)~~
文摘To rapidly obtain high-quality genomic DNA from Chenopodium quinoa Willd, the genomic DAN in different tissues （leaves, stems and roots） of Chenopodi- um quinoa Willd was extracted by modified CTAB method, SDS method and high- salt Iow-pH method, respectively. The quality and yield of extracted DNA was deter- mined using agarose gel electrophoresis and UV spectrophotometry. At the same time, the PCR-SSR and SSCP molecular detection was also performed. The results showed that the gel test strips, without obvious decomposition, of all the extraction methods were relatively obvious; the genomic DNA yield extracted by modified CTAB method was highest, followed by that by SDS method, and the genomic DNA extracted by high-salt Iow-pH method was lowest： the genomic DNA yields extracted by different methods from Chenopodium quinoa Wiltd leaves were all high- er than those from roots and stems; the quality of Chenopodium quinoa Willd ge- nomic DNA extracted by modified CTAB method and high-salt Iow-pH method was better, and polyphenols, polysaccharides and other impurities were removed more completely. The PCR-SSR and SSCP detection results showed that the genomic DNA extracted by different methods from different tissues of Chenopodium quinoa Willd all could be better amplified, and high-quality strips could be obtained. So the Chenopodium quinoa Willd genomic DNA extracted by the three methods all can be used for subsequent molecular biology research.
基金Supported by Natural Science Fund of Ningxia(NZ0866)~~
文摘[Objective] The aim of this study was to observe the agronomic,yield and quality traits of Ningchun 4 and its parents under the same environmental conditions,as well as to carry out molecular detection on the background genes distribution of the corresponding traits.[Method]Ningchun 4 and its parents Sonora 64,Hongtu,Abbondanza,Quality were used as materials to detect the agronomic and quality traits,as well as to analyze the genetic variation laws by molecular determination method.[Result]Ningchun 4 had inherited the advantages of bigger spike,red and hard grain from Sonora 64 and higher 1 000-grain weight from Hongtu.However,it had also inherited the disadvantages of late-maturing from Sonora 64 and lower tillering ability from Hongtu;the grain quality of Ningchun 4 was slightly lower than the Sonora 64;Ningchun 4 had high quality subunit of 5＋10,which had good dough rheological properties.[Conclusion]Ningchun 4 had inherited the long photoperiod characteristics and no-resistant to slow-leaf rusting and stripe rust characteristics from Hongtu and low PPO activity,high yellow pigments content from parents.
基金State Key Program for Basic Research Grants (2006CB101801)
文摘White spot syndrome virus (WSSV),Taura syndrome virus (TSV) and Infectious hypodermal and haematopoietic necrosis virus (IHHNV) are three shrimp viruses responsible for major pandemics affecting the shrimp farming industry. Shrimps samples were collected from 12 farms in Zhejiang province,China,in 2008 and analyzed by PCR to determine the prevalence of these viruses. From the 12 sampling locations,8 farms were positive for WSSV,8 for IHHNV and 6 for both WSSV and IHHNV. An average percentage of 57.4% of shrimp individuals were infected with WSSV,while 49.2% were infected with IHHNV. A high prevalence of co-infection with WSSV and IHHNV among samples was detected from the following samples:Bingjiang (93.3%),liuao (66.7%),Jianshan (46.7%) and Xianxiang (46.7%). No samples exhibited evidence of infection with TSV in collected samples. This study provides comprehensive information of the prevalence of three shrimp viruses in Zhejiang and may be helpful for disease prevention control in this region.
基金supported by grants from the Special Fund for Agro-scientific Research in the Public Interest, China (201203076)the Opening Fund of the State Key Laboratory for Biology of Plant Diseases and Insect Pests, China (SKLOF201518)
文摘The common fig（Ficus carica L.）was one of the earliest horticultural crops to be domesticated.A number of different viruses can infect fig trees including Fig mosaic virus（FMV）that has been detected in several commercial fig trees in Xinjiang,China.However,the distribution of FMV and other fig-infecting viruses in China remains unknown.In the present study,a sample from an ancient fig tree growing in Xinjiang was investigated by electron microscopy（EM）followed by PCR/RT-PCR,and FMV,Fig badnavirus 1（FBV-1）and Fig leaf mottle-associated virus 1（FLMaV-1）were detected.Fig leaf samples（252）from commercial orchards across China were subjected to PCR/RT-PCR,and FMV,FBV-1 and Fig fleck-associated virus（FFka V）were relatively abundant（44.4,48.4 and 44%,respectively）,while FLMaV-1 and Fig mild mottle-associated virus（FMMa V）were much scarcer（5.6 and 0.4%,respectively）,and FLMaV-2,Fig cryptic virus（FCV）,and Fig latent virus（FLV）were not detected.The presence of disease-causing viruses in fig trees presents a significant challenge for fig producers in China.This study may help to promote actions aimed at controlling fig viruses,especially FMV.
