【目的】建立基于鸡毒支原体种特异性粘附蛋白编码基因pvpA的实时荧光定量PCR检测方法。【方法】根据GenBank公布的不同国家和地区鸡毒支原体pvpA基因序列,在高度保守区域设计一对引物。以临床分离鸡毒支原体RC1为模板扩增pvpA基因,将...【目的】建立基于鸡毒支原体种特异性粘附蛋白编码基因pvpA的实时荧光定量PCR检测方法。【方法】根据GenBank公布的不同国家和地区鸡毒支原体pvpA基因序列,在高度保守区域设计一对引物。以临床分离鸡毒支原体RC1为模板扩增pvpA基因,将其连接到PMD19-T载体,转化至大肠杆菌DH5α中,经PCR和酶切鉴定并测序验证后得到阳性重组质粒rPvpA90。以rPvpA90为模板建立SYBR Green I荧光定量的标准曲线和溶点曲线,并进行特异性,灵敏性,重复性及临床样本检测试验,评价该方法的可行性。【结果】所建立的荧光定量PCR标准曲线循环阈值与模板浓度呈良好的线性关系,溶点曲线特异,相关系数为0.990。最低检测限为72拷贝/20μL,其敏感性比常规PCR至少高100倍;无论是对不同病原DNA单模板还是几种病原DNA混合模板进行扩增,该方法都呈现很好的特异性;重复性试验中,批内和批间变异系数均小于2%,表明该方法重现性好;临床样本的检测结果表明所建立的荧光定量PCR检测方法的检测率明显高于常规PCR方法。【结论】本研究初步建立了基于种特异性基因pvpA的鸡毒支原体荧光定量PCR方法,为养禽场诊断和监测鸡毒支原体病原提供一种新的特异、灵敏的方法。展开更多
To evaluate the supplementary blue light intensity on growth and health-promoting compounds in pak choi（Brassica campestris ssp.chinensis var.communis）,four blue light intensity treatments（T0,T50,T100 and T150 indi...To evaluate the supplementary blue light intensity on growth and health-promoting compounds in pak choi（Brassica campestris ssp.chinensis var.communis）,four blue light intensity treatments（T0,T50,T100 and T150 indicate 0,50,100,and 150μmol m^（-2） s^（-1）,respectively）were applied 10 days before harvest under greenhouse conditions.Both of cultivars（greenand red-leaf pak choi）under T50 had the highest yield,content of chlorophyll and sugars.With light intensity increasing,antioxidant compounds（vitamin C and carotenoids）significantly increased,while nitrate content showed an opposite trend.The health-promoting compounds（phenolics,flavonoids,anthocyanins,and glucosinolates）were significantly higher under supplementary light treatment than T0,so as the antioxidant capacity（2,2-diphenyl-1-picrylhydrazyl and ferric-reducing antioxidant power）.The species-specific differences in photosynthetic pigment and health-promoting compounds was found in green-and red-leaf pak choi.T50 treatment could be used for yield improvement,whereas T100 treatment could be applied for quality improvement.Results showed that blue light intensity can regulate the accumulation of biomass,morphology and health-promoting compounds in pak choi under greenhouse conditions.展开更多
Scutellaria baicalensis(S.baicalensis)and Scutellaria barbata(S.barbata)are common medicinal plants of the Lamiaceae family.Both produce specific flavonoid compounds,including baicalein,scutellarein,norwogonin,and wog...Scutellaria baicalensis(S.baicalensis)and Scutellaria barbata(S.barbata)are common medicinal plants of the Lamiaceae family.Both produce specific flavonoid compounds,including baicalein,scutellarein,norwogonin,and wogonin,as well as their glycosides,which exhibit antioxidant and antitumor activities.Here,we report chromosome-level genome assemblies of S.baicalensis and S.barbata with quantitative chromosomal variation(2 n=18 and 2 n=26,respectively).The divergence of S.baicalensis and S.barbata occurred far earlier than previously reported,and a whole-genome duplication(WGD)event was identified.The insertion of long terminal repeat elements after speciation might be responsible for the observed chromosomal expansion and rearrangement.Comparative genome analysis of the congeneric species revealed the species-specific evolution of chrysin and apigenin biosynthetic genes,such as the S.baicalensis-specific tandem duplication of genes encoding phenylalanine ammonia lyase and chalcone synthase,and the S.barbata-specific duplication of genes encoding 4-Co A ligase.In addition,the paralogous duplication,colinearity,and expression diversity of CYP82 D subfamily members revealed the functional divergence of genes encoding flavone hydroxylase between S.baicalensis and S.barbata.Analyzing these Scutellaria genomes reveals the common and species-specific evolution of flavone biosynthetic genes.Thus,these findings would facilitate the development of molecular breeding and studies of biosynthesis and regulation of bioactive compounds.展开更多
Tylenchulus semipenetrans is the most economically important and widespread nematode pest of citrus in China.rDNA-ITS of 14 populations of T.semipenetrans which were collected from different citrus groves or Chinese ...Tylenchulus semipenetrans is the most economically important and widespread nematode pest of citrus in China.rDNA-ITS of 14 populations of T.semipenetrans which were collected from different citrus groves or Chinese fir（Cunninghamia lanceolata） plantations in China were amplified and sequenced.The species-specific primers were designed for the first time to diagnosis T.semipenetrans based on the sequences of rDNA-ITS regions of geographic population above.The primers were sensitive to amplify the expected band size（297 bp） from DNA template of a single second-stage juvenile（J2） or different life stages of T.semipenetrans.No specific band was amplified from 15 non-target nematode species which were commonly found in citrus groves.Specificity and reliability of the primers were validated by further PCR amplification of 16 extra populations of T.semipenetrans collected from 4 provinces of China.The primers successfully detected a single J2 of T.semipenetrans within a whole nematode community comprising a large numbers of non-target nematode.The developed diagnostic technique can be used for accurate identification of T.semipenetrans and also as a decision tool for nematode management for citrus or Chinese fir in China.展开更多
A SYBR Green I real-time PCR assay was developed to detect and quantify Plasmodiophora brassicae ribosomal DNA（rDNA） and internal transcribed spacer（ITS）.A pair of primers PBF1/PBR1 was designed based on the conse...A SYBR Green I real-time PCR assay was developed to detect and quantify Plasmodiophora brassicae ribosomal DNA（rDNA） and internal transcribed spacer（ITS）.A pair of primers PBF1/PBR1 was designed based on the conservative region of rDNA-ITS of P.brassicae.The positive plasmid pB12 was obtained and used as the template to create standard curve.The specificity,sensitivity,and reproducibility of real-time PCR were evaluated respectively.Naturally and artificially infested soil samples containing different concentrations of P.brassicae were detected.The results demonstrated that standard curve established by recombinant plasmid was shown a fine linear relationship between threshold cycle and template concentration.The melting curve was specific with the correlation coefficient of 0.995 and that the amplification efficiency was 93.8%.The detection limit of P.brassicae genomic DNA was approximately 40 copies per 25 μL.The sensitivity of the assay was at least 100-fold higher than conventional PCR.Only DNA from P.brassicae could be amplified and detected using this assay,suggesting the highly specific of this assay.The coefficient of variation was less than 3%,indicating the PCR method revealed high reproducibility.The detection limit in soil samples corresponded to 1 000 resting spores g-1soil.Bait plants were used to validate the real-time PCR assay.This developed real-time PCR assay allows for fast and sensitive detection of P.brassicae in soil and should be useful in disease management and pest interception so as to prevent further spread of P.brassicae.展开更多
Phyllosticta species associated with diseases of four commercial Citrus species grown in China are reported.Totally,496 Phyllosticta strains were isolated from mandarins(Citrus reticulata),pomeloes(C.maxima),oranges(C...Phyllosticta species associated with diseases of four commercial Citrus species grown in China are reported.Totally,496 Phyllosticta strains were isolated from mandarins(Citrus reticulata),pomeloes(C.maxima),oranges(C.sinensis)and lemons(C.limon)in the main citrus producing regions across China,and 74 strains were selected for phylogenetic analysis.Analyses inferred from the sequences of internal transcribed spacer region(ITS1,5.8S nrDNA and ITS2),partial translation elongation factor 1-alpha(TEF1)and partial actin gene(ACT),showed these representative Phyllosticta isolates clustered in four distinct clades corresponding to three known,and one undescribed species.The newly resolved taxon,Phyllosticta citrichinaensis was isolated from leaves and fruits of all four Citrus species and is introduced in this paper.This taxon caused minor damage,showing irregular spots or freckles.Phyllosticta citriasiana,associated with tan spot of pomeloes,was isolated only from pomeloes,and never from lemons,mandarins and oranges.Phyllosticta citricarpa,the citrus black spot pathogen,which is presently subjected to phytosanitary legislation in the EU and United States,was isolated from lemons,mandarins and oranges,but never from pomeloes.The isolates of P.citricarpa clustered in two subclades,one from mandarins,the other from oranges and lemons.P.capitalensis was isolated from all four Citrus species as an endophyte,causing false melanose,or together with P.citricarpa or P.citriasiana.Morphological,cultural and biochemical characters were consistent with the results of phylogenetic analysis.In addition,a specific primer pair Pca8/ITS4 was designed and selected,and its corresponding PCR procedure was developed for the detection of P.citriasiana in this study.展开更多
The inflection point is an important feature of sigmoidal height-diameter(H-D)models.It is often cited as one of the properties favoring sigmoidal model forms.However,there are very few studies analyzing the inflectio...The inflection point is an important feature of sigmoidal height-diameter(H-D)models.It is often cited as one of the properties favoring sigmoidal model forms.However,there are very few studies analyzing the inflection points of H-D models.The goals of this study were to theoretically and empirically examine the behaviors of inflection points of six common H-D models with a regional dataset.The six models were the Wykoff(WYK),Schumacher(SCH),Curtis(CUR),HossfeldⅣ(HOS),von Bertalanffy-Richards(VBR),and Gompertz(GPZ)models.The models were first fitted in their base forms with tree species as random effects and were then expanded to include functional traits and spatial distribution.The distributions of the estimated inflection points were similar between the two-parameter models WYK,SCH,and CUR,but were different between the threeparameter models HOS,VBR,and GPZ.GPZ produced some of the largest inflection points.HOS and VBR produced concave H-D curves without inflection points for 12.7%and 39.7%of the tree species.Evergreen species or decreasing shade tolerance resulted in larger inflection points.The trends in the estimated inflection points of HOS and VBR were entirely opposite across the landscape.Furthermore,HOS could produce concave H-D curves for portions of the landscape.Based on the studied behaviors,the choice between two-parameter models may not matter.We recommend comparing seve ral three-parameter model forms for consistency in estimated inflection points before deciding on one.Believing sigmoidal models to have inflection points does not necessarily mean that they will produce fitted curves with one.Our study highlights the need to integrate analysis of inflection points into modeling H-D relationships.展开更多
本文采用苹果属过敏原蛋白的mal d 4.02基因,桃属微卫星标记MA023a分别作为苹果汁和桃汁特异性引物,并以植物高度保守的叶绿体AccD基因作为内标物,利用PCR检测方法对两种果汁进行种属鉴定。结果表明,特异性PCR检测方法可以准确的对苹果...本文采用苹果属过敏原蛋白的mal d 4.02基因,桃属微卫星标记MA023a分别作为苹果汁和桃汁特异性引物,并以植物高度保守的叶绿体AccD基因作为内标物,利用PCR检测方法对两种果汁进行种属鉴定。结果表明,特异性PCR检测方法可以准确的对苹果和桃产品进行种属鉴定,苹果、桃属特异性PCR检测方法的最低检测限分别为5、10ng/μL,此方法能快速、准确对苹果汁和桃汁种类进行定性鉴定和掺假检测。展开更多
Objective:As an important food therapy product with traditional Chinese medicine(TCM) applications,donkey-hide gelatin(Asini Corii Colla,ACC) has been used for thousands of years.However,till now few effective strateg...Objective:As an important food therapy product with traditional Chinese medicine(TCM) applications,donkey-hide gelatin(Asini Corii Colla,ACC) has been used for thousands of years.However,till now few effective strategy had been proposed to distinguish ACC from other animal hide gelatins,especially closely related horse-and mule-hide gelatins,which was an embarrassment of ACC quality control.Methods:Combined mass spectrometry and bioinformatic methods have been applied to identify and verify two ACC-specific peptides(Pep-1 and Pep-2) capable of distinguishing ACC from other closely related animal gelatins with high selectivity.Results:It confirmed that these two peptides could be not only used for distinguishing ACC from highly homologous horse-hide and mule-hide gelatins as well as other animal hide gelatins.Conclusion:The present study provides a simple method for species-specific peptides discovery,which can be used for assessing the quality of animal gelatin products,and ensure they are authenticable and traceable.展开更多
The globally invasive cassava mealybug Phenacoccus manihoti Matile-Ferrero is a pernicious pest of cassava,and its recent introduction into Asia has raised considerable alarm.To slow or prevent further invasion,an acc...The globally invasive cassava mealybug Phenacoccus manihoti Matile-Ferrero is a pernicious pest of cassava,and its recent introduction into Asia has raised considerable alarm.To slow or prevent further invasion,an accurate,simple,and developmental-stage-independent detection method for P.manihoti is required.In the present study,a PCR method based on a species-specific mitochondrial DNA cytochrome oxidase I(SS-COI)marker was developed for rapid identification of P.manihoti.One pair of SS-COI primers(PMSSZW-1F and PMSSZW-1R)was designed based on sequence variations in the COI gene among P.manihoti and related mealybug species.Specificity of the primer pair was validated on 21 closely related species.Sensitivity tests were performed on four immature developmental stages and female adults.Efficacy tests demonstrated that at the relatively low concentration of(135.2±14.7)pgresuspended DNA,the specific fragment was detected in all replicates.Furthermore,the SS-COI primer pair was assayed on three populations of P.manihoti from major exporting countries of cassava.The PCR assay was proved to be a rapid,simple,and reliable molecular measure for the identification of P.manihoti.This tool will be useful for quarantine,monitoring,and management of this invasive pest.展开更多
Botryosphaeriaceae species are important causal agents of blueberry stem blight worldwide. Blueberry stem blight has become an important disease, potentially affecting the quality and production of blueberries in Chin...Botryosphaeriaceae species are important causal agents of blueberry stem blight worldwide. Blueberry stem blight has become an important disease, potentially affecting the quality and production of blueberries in China. It is difficult and time-consuming to identify at the species level using morphological methods. The aim of this study was to develop polymerase chain reaction（PCR） assays for the diagnosis and early detection of latent infections of blueberry stems by Botryosphaeria spp. Species-specific primers, based on the ribosomal DNA internal transcribed spacer region and β-tubulin gene, were designed and selected for use in PCR assays. Three primer pairs, Lt347-F/R for Lasiodiplodia theobromae, Np304-F/R for Neofusicoccum parvum and FaF/Bt2b for Botryosphaeria dothidea, successfully amplified specific PCR fragments of different sizes on pure cultures or from blueberry stems inoculated and naturally infected blueberry plants with three pathogens, respectively. These primers did not amplify any PCR fragments from other blueberry stem disease-associated pathogens, such as Phomopsis spp. and Pestalotiopsis spp. This PCR protocol could detect as low as 1 00 pg to 1 ng of purified fungal DNA. This PCR-based protocol could be used for the diagnosis and detection of these pathogens from pure cultures or from infected blueberry plants.展开更多
Providing nest-boxes as surrogate tree cavities can be of great importance to increase the breeding populations of cavity-nesting birds in managed forests.However,the exact placement of nest-boxes should be taken into...Providing nest-boxes as surrogate tree cavities can be of great importance to increase the breeding populations of cavity-nesting birds in managed forests.However,the exact placement of nest-boxes should be taken into consideration to enhance their occupancy according to species-specific preferences.In this study,we investigated which factors can better predict nest-box occupancy by the Great Tit(Parus major)in eucalypt plantations.We used generalised linear mixed-effects models to analyse the influence of topography,nest-box positioning,vegetation cover and landscape variables on three-year occupancy records from 80 newly provided nest-boxes.Non-random patterns of nest-box occupancy were found with respect to all categories except topography.Results suggest that Great Tits prefer to occupy high-placed nest-boxes,close to areas that can provide them with supplementary resources either within or in the vicinity of the stand(i.e.,trees other than eucalypts,riparian vegetation,and large patches of adjacent habitats).Overall,this study provides important recommendations for nest-box placement and spatial distribution in managed forests and enhances the potential of nest-box interventions as a biodiversity offset and management tool.展开更多
文摘【目的】建立基于鸡毒支原体种特异性粘附蛋白编码基因pvpA的实时荧光定量PCR检测方法。【方法】根据GenBank公布的不同国家和地区鸡毒支原体pvpA基因序列,在高度保守区域设计一对引物。以临床分离鸡毒支原体RC1为模板扩增pvpA基因,将其连接到PMD19-T载体,转化至大肠杆菌DH5α中,经PCR和酶切鉴定并测序验证后得到阳性重组质粒rPvpA90。以rPvpA90为模板建立SYBR Green I荧光定量的标准曲线和溶点曲线,并进行特异性,灵敏性,重复性及临床样本检测试验,评价该方法的可行性。【结果】所建立的荧光定量PCR标准曲线循环阈值与模板浓度呈良好的线性关系,溶点曲线特异,相关系数为0.990。最低检测限为72拷贝/20μL,其敏感性比常规PCR至少高100倍;无论是对不同病原DNA单模板还是几种病原DNA混合模板进行扩增,该方法都呈现很好的特异性;重复性试验中,批内和批间变异系数均小于2%,表明该方法重现性好;临床样本的检测结果表明所建立的荧光定量PCR检测方法的检测率明显高于常规PCR方法。【结论】本研究初步建立了基于种特异性基因pvpA的鸡毒支原体荧光定量PCR方法,为养禽场诊断和监测鸡毒支原体病原提供一种新的特异、灵敏的方法。
基金supported by the National Key Research and Development Program of China (2017YFD0701500)the Teamwork Projects Funded by Guangdong Natural Science Foundation, China (S2013030012842)+1 种基金the Guangdong Provincial Science & Technology Project, China (2015A020209146, 2015B090903074)the Guangzhou Science & Technology Project, China (201605030005, 201704020058)
文摘To evaluate the supplementary blue light intensity on growth and health-promoting compounds in pak choi（Brassica campestris ssp.chinensis var.communis）,four blue light intensity treatments（T0,T50,T100 and T150 indicate 0,50,100,and 150μmol m^（-2） s^（-1）,respectively）were applied 10 days before harvest under greenhouse conditions.Both of cultivars（greenand red-leaf pak choi）under T50 had the highest yield,content of chlorophyll and sugars.With light intensity increasing,antioxidant compounds（vitamin C and carotenoids）significantly increased,while nitrate content showed an opposite trend.The health-promoting compounds（phenolics,flavonoids,anthocyanins,and glucosinolates）were significantly higher under supplementary light treatment than T0,so as the antioxidant capacity（2,2-diphenyl-1-picrylhydrazyl and ferric-reducing antioxidant power）.The species-specific differences in photosynthetic pigment and health-promoting compounds was found in green-and red-leaf pak choi.T50 treatment could be used for yield improvement,whereas T100 treatment could be applied for quality improvement.Results showed that blue light intensity can regulate the accumulation of biomass,morphology and health-promoting compounds in pak choi under greenhouse conditions.
基金the National Key R&D Program of China(Grant No.2019YFC1711100)the National Natural Science Foundation of China(Grant No.31700264)the Chinese Academy of Medical Sciences(CAMS)Innovation Fund for Medical Sciences(CIFMS)(Grant No.2016-I2M3-016)。
文摘Scutellaria baicalensis(S.baicalensis)and Scutellaria barbata(S.barbata)are common medicinal plants of the Lamiaceae family.Both produce specific flavonoid compounds,including baicalein,scutellarein,norwogonin,and wogonin,as well as their glycosides,which exhibit antioxidant and antitumor activities.Here,we report chromosome-level genome assemblies of S.baicalensis and S.barbata with quantitative chromosomal variation(2 n=18 and 2 n=26,respectively).The divergence of S.baicalensis and S.barbata occurred far earlier than previously reported,and a whole-genome duplication(WGD)event was identified.The insertion of long terminal repeat elements after speciation might be responsible for the observed chromosomal expansion and rearrangement.Comparative genome analysis of the congeneric species revealed the species-specific evolution of chrysin and apigenin biosynthetic genes,such as the S.baicalensis-specific tandem duplication of genes encoding phenylalanine ammonia lyase and chalcone synthase,and the S.barbata-specific duplication of genes encoding 4-Co A ligase.In addition,the paralogous duplication,colinearity,and expression diversity of CYP82 D subfamily members revealed the functional divergence of genes encoding flavone hydroxylase between S.baicalensis and S.barbata.Analyzing these Scutellaria genomes reveals the common and species-specific evolution of flavone biosynthetic genes.Thus,these findings would facilitate the development of molecular breeding and studies of biosynthesis and regulation of bioactive compounds.
基金supported by the National Natural Science Foundation of China (30700526)the Postdoctoral Science Foundation of China (55920)the Science Foundation of the Fujian Province,China (2009N0013)
文摘Tylenchulus semipenetrans is the most economically important and widespread nematode pest of citrus in China.rDNA-ITS of 14 populations of T.semipenetrans which were collected from different citrus groves or Chinese fir（Cunninghamia lanceolata） plantations in China were amplified and sequenced.The species-specific primers were designed for the first time to diagnosis T.semipenetrans based on the sequences of rDNA-ITS regions of geographic population above.The primers were sensitive to amplify the expected band size（297 bp） from DNA template of a single second-stage juvenile（J2） or different life stages of T.semipenetrans.No specific band was amplified from 15 non-target nematode species which were commonly found in citrus groves.Specificity and reliability of the primers were validated by further PCR amplification of 16 extra populations of T.semipenetrans collected from 4 provinces of China.The primers successfully detected a single J2 of T.semipenetrans within a whole nematode community comprising a large numbers of non-target nematode.The developed diagnostic technique can be used for accurate identification of T.semipenetrans and also as a decision tool for nematode management for citrus or Chinese fir in China.
基金supported by the emarked fund for Moden Agro-Industry Technology Research System, China (CARS25)the National Natural Science Foundation of China (31201473)the Key Laboratory of Biology and Genetic Improvement of Horticulture Crops, Ministry of Agriculture, China
文摘A SYBR Green I real-time PCR assay was developed to detect and quantify Plasmodiophora brassicae ribosomal DNA（rDNA） and internal transcribed spacer（ITS）.A pair of primers PBF1/PBR1 was designed based on the conservative region of rDNA-ITS of P.brassicae.The positive plasmid pB12 was obtained and used as the template to create standard curve.The specificity,sensitivity,and reproducibility of real-time PCR were evaluated respectively.Naturally and artificially infested soil samples containing different concentrations of P.brassicae were detected.The results demonstrated that standard curve established by recombinant plasmid was shown a fine linear relationship between threshold cycle and template concentration.The melting curve was specific with the correlation coefficient of 0.995 and that the amplification efficiency was 93.8%.The detection limit of P.brassicae genomic DNA was approximately 40 copies per 25 μL.The sensitivity of the assay was at least 100-fold higher than conventional PCR.Only DNA from P.brassicae could be amplified and detected using this assay,suggesting the highly specific of this assay.The coefficient of variation was less than 3%,indicating the PCR method revealed high reproducibility.The detection limit in soil samples corresponded to 1 000 resting spores g-1soil.Bait plants were used to validate the real-time PCR assay.This developed real-time PCR assay allows for fast and sensitive detection of P.brassicae in soil and should be useful in disease management and pest interception so as to prevent further spread of P.brassicae.
基金This work was supported by the earmarked fund for Modern Agro-industry Technology Research System(MATRS)of China,the National Foundation of Natural Science of China(31071649)The Global Research Network for Fungal Biology and King Saud University are thanked for supporting this research.MFLU awarded grant No 53101020017 to study the genus Phyllosticta in northern Thailand and the National Research Council of Thailand awarded grant No 54201020004 to study the genus Phyllosticta in Thailand.
文摘Phyllosticta species associated with diseases of four commercial Citrus species grown in China are reported.Totally,496 Phyllosticta strains were isolated from mandarins(Citrus reticulata),pomeloes(C.maxima),oranges(C.sinensis)and lemons(C.limon)in the main citrus producing regions across China,and 74 strains were selected for phylogenetic analysis.Analyses inferred from the sequences of internal transcribed spacer region(ITS1,5.8S nrDNA and ITS2),partial translation elongation factor 1-alpha(TEF1)and partial actin gene(ACT),showed these representative Phyllosticta isolates clustered in four distinct clades corresponding to three known,and one undescribed species.The newly resolved taxon,Phyllosticta citrichinaensis was isolated from leaves and fruits of all four Citrus species and is introduced in this paper.This taxon caused minor damage,showing irregular spots or freckles.Phyllosticta citriasiana,associated with tan spot of pomeloes,was isolated only from pomeloes,and never from lemons,mandarins and oranges.Phyllosticta citricarpa,the citrus black spot pathogen,which is presently subjected to phytosanitary legislation in the EU and United States,was isolated from lemons,mandarins and oranges,but never from pomeloes.The isolates of P.citricarpa clustered in two subclades,one from mandarins,the other from oranges and lemons.P.capitalensis was isolated from all four Citrus species as an endophyte,causing false melanose,or together with P.citricarpa or P.citriasiana.Morphological,cultural and biochemical characters were consistent with the results of phylogenetic analysis.In addition,a specific primer pair Pca8/ITS4 was designed and selected,and its corresponding PCR procedure was developed for the detection of P.citriasiana in this study.
文摘The inflection point is an important feature of sigmoidal height-diameter(H-D)models.It is often cited as one of the properties favoring sigmoidal model forms.However,there are very few studies analyzing the inflection points of H-D models.The goals of this study were to theoretically and empirically examine the behaviors of inflection points of six common H-D models with a regional dataset.The six models were the Wykoff(WYK),Schumacher(SCH),Curtis(CUR),HossfeldⅣ(HOS),von Bertalanffy-Richards(VBR),and Gompertz(GPZ)models.The models were first fitted in their base forms with tree species as random effects and were then expanded to include functional traits and spatial distribution.The distributions of the estimated inflection points were similar between the two-parameter models WYK,SCH,and CUR,but were different between the threeparameter models HOS,VBR,and GPZ.GPZ produced some of the largest inflection points.HOS and VBR produced concave H-D curves without inflection points for 12.7%and 39.7%of the tree species.Evergreen species or decreasing shade tolerance resulted in larger inflection points.The trends in the estimated inflection points of HOS and VBR were entirely opposite across the landscape.Furthermore,HOS could produce concave H-D curves for portions of the landscape.Based on the studied behaviors,the choice between two-parameter models may not matter.We recommend comparing seve ral three-parameter model forms for consistency in estimated inflection points before deciding on one.Believing sigmoidal models to have inflection points does not necessarily mean that they will produce fitted curves with one.Our study highlights the need to integrate analysis of inflection points into modeling H-D relationships.
文摘本文采用苹果属过敏原蛋白的mal d 4.02基因,桃属微卫星标记MA023a分别作为苹果汁和桃汁特异性引物,并以植物高度保守的叶绿体AccD基因作为内标物,利用PCR检测方法对两种果汁进行种属鉴定。结果表明,特异性PCR检测方法可以准确的对苹果和桃产品进行种属鉴定,苹果、桃属特异性PCR检测方法的最低检测限分别为5、10ng/μL,此方法能快速、准确对苹果汁和桃汁种类进行定性鉴定和掺假检测。
基金funded by the National Natural Science Foundation of China (No.81973450)the National Key R&D Program of China (No.2018YFC1706100)+4 种基金the Jiangsu Qinglan Projectthe Jiangsu “333” Projectthe Young Researchers Training Project of China Association of Traditional Chinese Medicine (No.QNRC2C14)the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (No.19KJB360020)the Open Project of Chinese Materia Medica First-Class Discipline of Nanjing University of Chinese Medicine (2020YLXK009)。
文摘Objective:As an important food therapy product with traditional Chinese medicine(TCM) applications,donkey-hide gelatin(Asini Corii Colla,ACC) has been used for thousands of years.However,till now few effective strategy had been proposed to distinguish ACC from other animal hide gelatins,especially closely related horse-and mule-hide gelatins,which was an embarrassment of ACC quality control.Methods:Combined mass spectrometry and bioinformatic methods have been applied to identify and verify two ACC-specific peptides(Pep-1 and Pep-2) capable of distinguishing ACC from other closely related animal gelatins with high selectivity.Results:It confirmed that these two peptides could be not only used for distinguishing ACC from highly homologous horse-hide and mule-hide gelatins as well as other animal hide gelatins.Conclusion:The present study provides a simple method for species-specific peptides discovery,which can be used for assessing the quality of animal gelatin products,and ensure they are authenticable and traceable.
基金supported by the National Key R&D Program of China(2017YFC1200600,2016YFC1201200 and 2015BAD08A16)the Science and Technology Innovation Program of CAAS(caascx-2013-2018-IAS)
文摘The globally invasive cassava mealybug Phenacoccus manihoti Matile-Ferrero is a pernicious pest of cassava,and its recent introduction into Asia has raised considerable alarm.To slow or prevent further invasion,an accurate,simple,and developmental-stage-independent detection method for P.manihoti is required.In the present study,a PCR method based on a species-specific mitochondrial DNA cytochrome oxidase I(SS-COI)marker was developed for rapid identification of P.manihoti.One pair of SS-COI primers(PMSSZW-1F and PMSSZW-1R)was designed based on sequence variations in the COI gene among P.manihoti and related mealybug species.Specificity of the primer pair was validated on 21 closely related species.Sensitivity tests were performed on four immature developmental stages and female adults.Efficacy tests demonstrated that at the relatively low concentration of(135.2±14.7)pgresuspended DNA,the specific fragment was detected in all replicates.Furthermore,the SS-COI primer pair was assayed on three populations of P.manihoti from major exporting countries of cassava.The PCR assay was proved to be a rapid,simple,and reliable molecular measure for the identification of P.manihoti.This tool will be useful for quarantine,monitoring,and management of this invasive pest.
基金supported financially by the National Natural Science Foundation of China (31301610)
文摘Botryosphaeriaceae species are important causal agents of blueberry stem blight worldwide. Blueberry stem blight has become an important disease, potentially affecting the quality and production of blueberries in China. It is difficult and time-consuming to identify at the species level using morphological methods. The aim of this study was to develop polymerase chain reaction（PCR） assays for the diagnosis and early detection of latent infections of blueberry stems by Botryosphaeria spp. Species-specific primers, based on the ribosomal DNA internal transcribed spacer region and β-tubulin gene, were designed and selected for use in PCR assays. Three primer pairs, Lt347-F/R for Lasiodiplodia theobromae, Np304-F/R for Neofusicoccum parvum and FaF/Bt2b for Botryosphaeria dothidea, successfully amplified specific PCR fragments of different sizes on pure cultures or from blueberry stems inoculated and naturally infected blueberry plants with three pathogens, respectively. These primers did not amplify any PCR fragments from other blueberry stem disease-associated pathogens, such as Phomopsis spp. and Pestalotiopsis spp. This PCR protocol could detect as low as 1 00 pg to 1 ng of purified fungal DNA. This PCR-based protocol could be used for the diagnosis and detection of these pathogens from pure cultures or from infected blueberry plants.
基金co-financed by Funda?ao para a Ciencia e a Tecnologia(FCT)the European Regional Development Fund(FEDER)through Portugal 2020 Competitiveness and Internationalization Operational Programme(POCI),reference POCI-01-0145-FEDER-030250 and PTDC/ASP-SIL/30250/2017-TOPDEVIL+1 种基金the R&D Unit Centre for Functional Ecology-Science for People and the Planet(CFE),with reference UIDB/04004/2020,financed by FCT/MCTES through national funds(PIDDAC)FCT/MCTES also funded L.P.S.with contract CEECIND/02064/2017。
文摘Providing nest-boxes as surrogate tree cavities can be of great importance to increase the breeding populations of cavity-nesting birds in managed forests.However,the exact placement of nest-boxes should be taken into consideration to enhance their occupancy according to species-specific preferences.In this study,we investigated which factors can better predict nest-box occupancy by the Great Tit(Parus major)in eucalypt plantations.We used generalised linear mixed-effects models to analyse the influence of topography,nest-box positioning,vegetation cover and landscape variables on three-year occupancy records from 80 newly provided nest-boxes.Non-random patterns of nest-box occupancy were found with respect to all categories except topography.Results suggest that Great Tits prefer to occupy high-placed nest-boxes,close to areas that can provide them with supplementary resources either within or in the vicinity of the stand(i.e.,trees other than eucalypts,riparian vegetation,and large patches of adjacent habitats).Overall,this study provides important recommendations for nest-box placement and spatial distribution in managed forests and enhances the potential of nest-box interventions as a biodiversity offset and management tool.
