Late blight caused by the oomycete Phytophthora infestans Montagne de Bary is a devastating disease on potato production. A total of six parameters affecting the PCR system in P. infestans were investigated based on t...Late blight caused by the oomycete Phytophthora infestans Montagne de Bary is a devastating disease on potato production. A total of six parameters affecting the PCR system in P. infestans were investigated based on the template DNA of the isolate HH06-23 and EST-SSR primer pair Pi08N. The results showed that the optimal annealing temperature was 63 ℃, and the optimum PCR system of EST-SSR was 25 ng template DNA, 0.5 mmol·L^-1 dNTPs, 2 μL 10×Buffer （Mg^2＋ free）, 1.75 mmol·L^-1 MgCl2, 15 pmol primer, and 1.2 U Taq DNA polymerase in total 20 μL reaction system. The PCR program was initial denaturation at 94℃ for 2 rain, followed by 35 cycles of 94℃ for 30 s, 63℃ for 30 s, and 72℃ for 30 s, then a final extension step was 72℃ for 7 min, and held at 4℃. In addition, using the optimal PCR system, a total of 20 isolates of P. infestans were used for testing the stability and polymorphism of the PCR amplification. The clarity and abundant polymorphism indicated that this system was stable and suitable for researching the genetic diversity ofP. infestans population.展开更多
A total of 104 isolates including two Korean isolates and three Japanese isolates of Phytophthora infestans collected from Heilongjiang and Jilin Provinces from 2006 to 2008 were used to determine their mating types,m...A total of 104 isolates including two Korean isolates and three Japanese isolates of Phytophthora infestans collected from Heilongjiang and Jilin Provinces from 2006 to 2008 were used to determine their mating types,metalaxyl resistance,and RAPD genotypes.All the isolates of P.infestans collected from Heilongjiang and Jilin Provinces belonged to A 1 mating type,and no A 2 mating type was detected.Frequencies of metalaxyl resistant isolates were 94.4%,47.8% and 75.0% in 2006,2007 and 2008,respectively.Accord...展开更多
The occurrence of sexual reproduction accelerates the population genetic variation of Phytophthora infestans and makes it more difficult to control.The systematic analysis of the differentiation of phenotype(mating ty...The occurrence of sexual reproduction accelerates the population genetic variation of Phytophthora infestans and makes it more difficult to control.The systematic analysis of the differentiation of phenotype(mating type and metalaxyl sensitivity)and genotype(mtDNA haplotype and SSR genotype)of 65 single oospore strains of P.infestans was carried out in this article.Five test strains were isolated from Heilongjiang Province and Mongolia Autonomous Region.The experiment results showed that the isolation ratio of metalaxyl resistance(MR:HR)of single oospore strains produced through the cross of medium resistance and high resistant parents was 18:13;the isolation ratio of the metalaxyl resistance(S:MR:HR)of single oospore strains produced through the cross of sensitive and high resistant parents was 4:12:7.The progenies of single oospore strains produced through self-fertility parents with medium resistance were all of the medium resistance.The mating types A1:A2 was greater than 1:1 in single oospore strains of the progenies,which did not conform to the Mendel's law of inheritance.All single oospore strains of the progenies inherited mitochondrial DNA fragments from only one parent.Sexual recombination of single oospore strains was verified by using two pairs of SSR primers(Pi4B and Pi4G).At the locus of Pi4B and Pi4G in the cross of KS-37 and KS-25,the separation frequencies of allele were 19:12 and 14:17,respectively.They produced two new genotype strains.This study could provide a basis for formulating disease control strategies.展开更多
基金Supported by Natural Science Foundation of Heilongjiang Province of China (C200622)
文摘Late blight caused by the oomycete Phytophthora infestans Montagne de Bary is a devastating disease on potato production. A total of six parameters affecting the PCR system in P. infestans were investigated based on the template DNA of the isolate HH06-23 and EST-SSR primer pair Pi08N. The results showed that the optimal annealing temperature was 63 ℃, and the optimum PCR system of EST-SSR was 25 ng template DNA, 0.5 mmol·L^-1 dNTPs, 2 μL 10×Buffer （Mg^2＋ free）, 1.75 mmol·L^-1 MgCl2, 15 pmol primer, and 1.2 U Taq DNA polymerase in total 20 μL reaction system. The PCR program was initial denaturation at 94℃ for 2 rain, followed by 35 cycles of 94℃ for 30 s, 63℃ for 30 s, and 72℃ for 30 s, then a final extension step was 72℃ for 7 min, and held at 4℃. In addition, using the optimal PCR system, a total of 20 isolates of P. infestans were used for testing the stability and polymorphism of the PCR amplification. The clarity and abundant polymorphism indicated that this system was stable and suitable for researching the genetic diversity ofP. infestans population.
基金Supported by Scientific Research Fund of Northeast Agricultural UniversityScience Foundation and Ministry of EducationReturned Overseas Students to Start-up Foundation (210150)
文摘A total of 104 isolates including two Korean isolates and three Japanese isolates of Phytophthora infestans collected from Heilongjiang and Jilin Provinces from 2006 to 2008 were used to determine their mating types,metalaxyl resistance,and RAPD genotypes.All the isolates of P.infestans collected from Heilongjiang and Jilin Provinces belonged to A 1 mating type,and no A 2 mating type was detected.Frequencies of metalaxyl resistant isolates were 94.4%,47.8% and 75.0% in 2006,2007 and 2008,respectively.Accord...
基金Supported by Heilongjiang Province Natural Science Foundation(C2016019)Cooperative Innovation System of Potato Modern Agricultural Industry Technology in Heilongjiang Province(HNWJZT201801)。
文摘The occurrence of sexual reproduction accelerates the population genetic variation of Phytophthora infestans and makes it more difficult to control.The systematic analysis of the differentiation of phenotype(mating type and metalaxyl sensitivity)and genotype(mtDNA haplotype and SSR genotype)of 65 single oospore strains of P.infestans was carried out in this article.Five test strains were isolated from Heilongjiang Province and Mongolia Autonomous Region.The experiment results showed that the isolation ratio of metalaxyl resistance(MR:HR)of single oospore strains produced through the cross of medium resistance and high resistant parents was 18:13;the isolation ratio of the metalaxyl resistance(S:MR:HR)of single oospore strains produced through the cross of sensitive and high resistant parents was 4:12:7.The progenies of single oospore strains produced through self-fertility parents with medium resistance were all of the medium resistance.The mating types A1:A2 was greater than 1:1 in single oospore strains of the progenies,which did not conform to the Mendel's law of inheritance.All single oospore strains of the progenies inherited mitochondrial DNA fragments from only one parent.Sexual recombination of single oospore strains was verified by using two pairs of SSR primers(Pi4B and Pi4G).At the locus of Pi4B and Pi4G in the cross of KS-37 and KS-25,the separation frequencies of allele were 19:12 and 14:17,respectively.They produced two new genotype strains.This study could provide a basis for formulating disease control strategies.
