This paper dealt with the SDS polyacrymide gel electrophoresis of proteins ahd analysis of the amino acids in the diseasedand healthy leaves with the same strain respectively, which were at the same side and height, o...This paper dealt with the SDS polyacrymide gel electrophoresis of proteins ahd analysis of the amino acids in the diseasedand healthy leaves with the same strain respectively, which were at the same side and height, of Paulownia catalpifolia, P. elongata, P. albiphloea and P. kauakamii respectively. The results indicated that the leaves of 4 species of Paulownia trees with witches' broom had one protein band, of which molecular weight was 12 kD, which did not appear in the healthy leaves free of phytoplasmas. Moreover, the protein quantity in the affected leaves was more than that in the healthy leaves free of phytoplasmas, At the same time, there was significant difference on the amino acids between the healthy and diseased leaves of P. catalpifolia and P. kawakamii. The amount of cystine in the affected leaves was higher than that in the healthy leaves, but the change of amount of phenylalanine in the affected and healthy leaves was contrary.These changes of proteins and amino acids in the leaves might be related to the witches' broom of the Parlounia trees.展开更多
Stem segments from diseased Paulownia tomentosaXP, fortunei and leaves from healthy control were transformed with the expression vector p438PRSI via Agrobacterium tumefaciens. The p438PRSI vector contained shiva-1 gen...Stem segments from diseased Paulownia tomentosaXP, fortunei and leaves from healthy control were transformed with the expression vector p438PRSI via Agrobacterium tumefaciens. The p438PRSI vector contained shiva-1 gene, which encodes an antibacterial peptide under the control of a CaMV35S promoter. The regenerated plants from transformed explants were planted in a greenhouse and nursery. PCR and Southern blotting analysis showed that the shiva-1 gene was successfully integrated into the Paulownia genome. Transcription of the integrated shiva-1 gene was confirmed by RT-PCR. Bioassay in the green house and phytoplasma DNA-dot blotting demonstrated that resistance to Paulownia witch＇s broom disease （PWB） increased significantly in shiva-l-transgenic Paulownia. Further investigations indicated that higher Shiva- 1 expression correlated with fewer phytoplasma and less symptoms in diseased transgenic Paulownia. Together, our findings strongly suggest that breeding shiva-1-Paulownia is an effective strategy to control PWB disease.展开更多
Paulownia witches’-broom disease which was caused by the paulownia witches’-broom phytoplasma(PaWB) is one of the severest diseases in paulownia production.Antigenic membrane protein(Amp) gene was cloned from Paulow...Paulownia witches’-broom disease which was caused by the paulownia witches’-broom phytoplasma(PaWB) is one of the severest diseases in paulownia production.Antigenic membrane protein(Amp) gene was cloned from Paulownia plants infected with Paulownia witches’-broom phytoplasma in Shaanxi Province.The gene was 696 bp in length,encoding a predicted protein of 231 amino acids.Homology analysis of sequence from PaWB and other 8 phytoplasmas of GenBank available showed that amp gene of PaWB was 100% identical to PaWB-Japan,and closely related to those of onion yellow(AY) and aster yellow(OY) in the same 16Sr Ⅰgroup with nucleotide acids sequences homology rate of 97%;However it had a lower nucleotide acids sequences homology rate of 37% with western X disease in 16Sr Ⅲ group.Prediction of protein structure showed that molecular weight of Amp was 24.7 ku and isoelectric point was 9.935.The Amp protein possessed acentral hydrophilic region and two hydrophobic transmembrane regions.There were several potential cleavage sites of signal peptide,the strongest cleavage site was at the end of Amp and there was no potential cleavage sites in the middle Amp.The information suggested that the protein should be of good antigenicity.展开更多
本实验以泡桐丛枝植原体为材料,运用差速离心和脉冲场电泳(Pulsed Field Gel Electrophoresis,PFGE),获得了无寄主DNA污染、完整的泡桐丛枝植原体染色体,并用Southern blot杂交进行了验证,建立了植原体全基因组的研究方法。同时对泡桐...本实验以泡桐丛枝植原体为材料,运用差速离心和脉冲场电泳(Pulsed Field Gel Electrophoresis,PFGE),获得了无寄主DNA污染、完整的泡桐丛枝植原体染色体,并用Southern blot杂交进行了验证,建立了植原体全基因组的研究方法。同时对泡桐丛枝植原体染色体进行内切酶I-CeuI不完全酶切,测定了染色体全长,并定位了2个非连锁的rRNA操纵子在染色体上的位置,完成了泡桐丛枝植原体部分物理图谱的构建。实验结果表明,泡桐丛枝植原体染色体全长约为1 100kb,2个rRNA操纵子在染色体上相距500～600kb。展开更多
文摘This paper dealt with the SDS polyacrymide gel electrophoresis of proteins ahd analysis of the amino acids in the diseasedand healthy leaves with the same strain respectively, which were at the same side and height, of Paulownia catalpifolia, P. elongata, P. albiphloea and P. kauakamii respectively. The results indicated that the leaves of 4 species of Paulownia trees with witches' broom had one protein band, of which molecular weight was 12 kD, which did not appear in the healthy leaves free of phytoplasmas. Moreover, the protein quantity in the affected leaves was more than that in the healthy leaves free of phytoplasmas, At the same time, there was significant difference on the amino acids between the healthy and diseased leaves of P. catalpifolia and P. kawakamii. The amount of cystine in the affected leaves was higher than that in the healthy leaves, but the change of amount of phenylalanine in the affected and healthy leaves was contrary.These changes of proteins and amino acids in the leaves might be related to the witches' broom of the Parlounia trees.
基金the National Natural Science Foundation of China，the President Foundation of the Chinese Academy of Sciences
文摘Stem segments from diseased Paulownia tomentosaXP, fortunei and leaves from healthy control were transformed with the expression vector p438PRSI via Agrobacterium tumefaciens. The p438PRSI vector contained shiva-1 gene, which encodes an antibacterial peptide under the control of a CaMV35S promoter. The regenerated plants from transformed explants were planted in a greenhouse and nursery. PCR and Southern blotting analysis showed that the shiva-1 gene was successfully integrated into the Paulownia genome. Transcription of the integrated shiva-1 gene was confirmed by RT-PCR. Bioassay in the green house and phytoplasma DNA-dot blotting demonstrated that resistance to Paulownia witch＇s broom disease （PWB） increased significantly in shiva-l-transgenic Paulownia. Further investigations indicated that higher Shiva- 1 expression correlated with fewer phytoplasma and less symptoms in diseased transgenic Paulownia. Together, our findings strongly suggest that breeding shiva-1-Paulownia is an effective strategy to control PWB disease.
文摘Paulownia witches’-broom disease which was caused by the paulownia witches’-broom phytoplasma(PaWB) is one of the severest diseases in paulownia production.Antigenic membrane protein(Amp) gene was cloned from Paulownia plants infected with Paulownia witches’-broom phytoplasma in Shaanxi Province.The gene was 696 bp in length,encoding a predicted protein of 231 amino acids.Homology analysis of sequence from PaWB and other 8 phytoplasmas of GenBank available showed that amp gene of PaWB was 100% identical to PaWB-Japan,and closely related to those of onion yellow(AY) and aster yellow(OY) in the same 16Sr Ⅰgroup with nucleotide acids sequences homology rate of 97%;However it had a lower nucleotide acids sequences homology rate of 37% with western X disease in 16Sr Ⅲ group.Prediction of protein structure showed that molecular weight of Amp was 24.7 ku and isoelectric point was 9.935.The Amp protein possessed acentral hydrophilic region and two hydrophobic transmembrane regions.There were several potential cleavage sites of signal peptide,the strongest cleavage site was at the end of Amp and there was no potential cleavage sites in the middle Amp.The information suggested that the protein should be of good antigenicity.
文摘本实验以泡桐丛枝植原体为材料,运用差速离心和脉冲场电泳(Pulsed Field Gel Electrophoresis,PFGE),获得了无寄主DNA污染、完整的泡桐丛枝植原体染色体,并用Southern blot杂交进行了验证,建立了植原体全基因组的研究方法。同时对泡桐丛枝植原体染色体进行内切酶I-CeuI不完全酶切,测定了染色体全长,并定位了2个非连锁的rRNA操纵子在染色体上的位置,完成了泡桐丛枝植原体部分物理图谱的构建。实验结果表明,泡桐丛枝植原体染色体全长约为1 100kb,2个rRNA操纵子在染色体上相距500～600kb。
