用CTAB法提取总DNA,合成位于18 S rDNA和26 S rDNA上的两条各20 bp的引物,通过PCR扩增ITS的全序列,对PCR产物直接测序,分别获得了硬蒺藜(Tribulus terrestrisL.)和软蒺藜(Atriplex centralasiaticaIljin)的核糖体RNA基因—rDNA内转录间...用CTAB法提取总DNA,合成位于18 S rDNA和26 S rDNA上的两条各20 bp的引物,通过PCR扩增ITS的全序列,对PCR产物直接测序,分别获得了硬蒺藜(Tribulus terrestrisL.)和软蒺藜(Atriplex centralasiaticaIljin)的核糖体RNA基因—rDNA内转录间隔区(ribosomal DNA internal transcribed spacer,rDNA-ITS)的序列643 bp和607bp,其碱基总差异率为36.16%,其中,ITS1的碱基差异率为55.81%;5.8 S的碱基差异率为6.59%;ITS2的碱基差异率为56.77%。这种差异,以及基因序列本身,为硬软蒺藜的区别和种质资源鉴定提供了分子依据。展开更多
Restriction fragment length polymorphism （RFLP） profiles of the amplified products of Internal Transcribed Spacer （ITS） region of rDNA using four restriction enzymes （Alul, Rsal, HinfI and HhaI） revealed distinc...Restriction fragment length polymorphism （RFLP） profiles of the amplified products of Internal Transcribed Spacer （ITS） region of rDNA using four restriction enzymes （Alul, Rsal, HinfI and HhaI） revealed distinctness of six Indian isolates of Steinernema one each from Maharashtra （IARI-EPN-mh）, Himachal Pradesh （IARI-EPN-hp）, Dehradun （IARI-EPN-dhdl）, Jharkhand （IARI-EPN-jhl） and two from Madhya Pradesh （IARI-EPN-bpll ＆ IARI-EPN-gwll）, when compared with the only native species Steinernema thermophilum. One of the restriction enzyme, Rsal could differentiate all the six species/strains from one another. The three restriction enzymes yielded patterns which were of diagnostic value but Rsal appeared to be the best diagnostic marker for differentiating these isolates. A tree constructed based upon the band sharing amongst the isolates, produced trichotomy which placed strains from Madhya Pradesh and Jharkhand in one group showing 94% homology, one strain from Bhopal （M.P） formed separate clade along with S. thermophilum with 72% similarity. These isolates, from Maharashtra, Himachal Pradesh and Dehradun, showed only 51% similarity with the S. thermophilum by forming separate clade.展开更多
Tomato seedlings damping-off is a limiting factor in commercial greenhouse production. To determine the causal agents of disease, sampling and fungal isolation were performed during 2012. Samples were collected from i...Tomato seedlings damping-off is a limiting factor in commercial greenhouse production. To determine the causal agents of disease, sampling and fungal isolation were performed during 2012. Samples were collected from infected seedlings growing in greenhouses in the Syrian coastal region. Isolation of fungi was done in the laboratories of the Agronomical Reaserch Center, in Lattakia and the molecular analyses were done in the Biotechnology Center at Tishreen University, Lattakia, Syria, during the years 2012, 2013. Eight isolates ofPythium sp. obtained were purified using hyphal tip method （named P1, P2, P3, P4, P5, P6, P7 and P8）. Isolates were morphologically identified by optical microscope, then molecularly Characterized using genus specific ITS primers. The results of morphological characterization of pathogenic species suggested the detection of Pythium aphanidermatum, P. ultimum. The analysis of DNAs from the different isolates with ITS primers, recognizing the inter transcript spacer of nuclear ribosomal DNA proved that the eight, isolates were belonging to the species P. ultimum. The complete sequences of ribosomal DNA internal transcribed spacers regions of selected isolates were determined and submitted to GenBank. The GenBank-BLAST homology search revealed P. ultimum as the most similar sequence （〉 96% identity） with GenBank entry AB355596.展开更多
文摘用CTAB法提取总DNA,合成位于18 S rDNA和26 S rDNA上的两条各20 bp的引物,通过PCR扩增ITS的全序列,对PCR产物直接测序,分别获得了硬蒺藜(Tribulus terrestrisL.)和软蒺藜(Atriplex centralasiaticaIljin)的核糖体RNA基因—rDNA内转录间隔区(ribosomal DNA internal transcribed spacer,rDNA-ITS)的序列643 bp和607bp,其碱基总差异率为36.16%,其中,ITS1的碱基差异率为55.81%;5.8 S的碱基差异率为6.59%;ITS2的碱基差异率为56.77%。这种差异,以及基因序列本身,为硬软蒺藜的区别和种质资源鉴定提供了分子依据。
文摘Restriction fragment length polymorphism （RFLP） profiles of the amplified products of Internal Transcribed Spacer （ITS） region of rDNA using four restriction enzymes （Alul, Rsal, HinfI and HhaI） revealed distinctness of six Indian isolates of Steinernema one each from Maharashtra （IARI-EPN-mh）, Himachal Pradesh （IARI-EPN-hp）, Dehradun （IARI-EPN-dhdl）, Jharkhand （IARI-EPN-jhl） and two from Madhya Pradesh （IARI-EPN-bpll ＆ IARI-EPN-gwll）, when compared with the only native species Steinernema thermophilum. One of the restriction enzyme, Rsal could differentiate all the six species/strains from one another. The three restriction enzymes yielded patterns which were of diagnostic value but Rsal appeared to be the best diagnostic marker for differentiating these isolates. A tree constructed based upon the band sharing amongst the isolates, produced trichotomy which placed strains from Madhya Pradesh and Jharkhand in one group showing 94% homology, one strain from Bhopal （M.P） formed separate clade along with S. thermophilum with 72% similarity. These isolates, from Maharashtra, Himachal Pradesh and Dehradun, showed only 51% similarity with the S. thermophilum by forming separate clade.
文摘Tomato seedlings damping-off is a limiting factor in commercial greenhouse production. To determine the causal agents of disease, sampling and fungal isolation were performed during 2012. Samples were collected from infected seedlings growing in greenhouses in the Syrian coastal region. Isolation of fungi was done in the laboratories of the Agronomical Reaserch Center, in Lattakia and the molecular analyses were done in the Biotechnology Center at Tishreen University, Lattakia, Syria, during the years 2012, 2013. Eight isolates ofPythium sp. obtained were purified using hyphal tip method （named P1, P2, P3, P4, P5, P6, P7 and P8）. Isolates were morphologically identified by optical microscope, then molecularly Characterized using genus specific ITS primers. The results of morphological characterization of pathogenic species suggested the detection of Pythium aphanidermatum, P. ultimum. The analysis of DNAs from the different isolates with ITS primers, recognizing the inter transcript spacer of nuclear ribosomal DNA proved that the eight, isolates were belonging to the species P. ultimum. The complete sequences of ribosomal DNA internal transcribed spacers regions of selected isolates were determined and submitted to GenBank. The GenBank-BLAST homology search revealed P. ultimum as the most similar sequence （〉 96% identity） with GenBank entry AB355596.
