[Objectives]To establish a simple,rapid and accurate method for identifying the genetic relationship of hybrid rice varieties to their parents.[Methods]Taking F1 hybrids Liangyou 336,Deliangyou Huazhan,and the parents...[Objectives]To establish a simple,rapid and accurate method for identifying the genetic relationship of hybrid rice varieties to their parents.[Methods]Taking F1 hybrids Liangyou 336,Deliangyou Huazhan,and the parents of Liangyou 336,i.e.,C815S(♀)and R336(♂),as experimental materials,the genetic relationship of the hybrid rice varieties to the parental materials was identified by way of PCR amplification with the 48 pairs of SSR primers of Protocol for Identification of Rice Varieties:SSR Marker Method(NY/T 1433-2014).[Results]The genetic relationship of the hybrid rice varieties could be determined by comparing the PCR amplification products of the mixed DNA of the parents and the DNA of the F1 hybrids.[Conclusions]This method not only reduced the number of samples required but also had a good visual effect and high accuracy.展开更多
The extraction of DNA is often the most time consuming and laborious step in high-throughput molecular genetic analysis and marker assisted selection （MAS） programs. A simple method for preparation of rice genomic D...The extraction of DNA is often the most time consuming and laborious step in high-throughput molecular genetic analysis and marker assisted selection （MAS） programs. A simple method for preparation of rice genomic DNA was developed. A small amount （1~50 mg） of leaf tissue of rice seedling, 500 pL of extraction buffer, and one steel bead were put into a 2-mL microcentrifuge tube. After vigorously mashing for 2 min, 5 μL of supernatant was directly applied to PCR amplification. Otherwise, the supematant was precipitated with two times volume of ethanol to obtain high quality genomic DNA. This method is simple, rapid, low cost, and reliable for PCR analysis. One person can manipulate as many as 96 samples for PCR in 10 min. It is especially suitable for genotyping of large number of samples.展开更多
A simple method to prepare of DNA template suitable for PCR amplification from filamentous fungi will be valuable for improving experimental efficiency.Here,a method was developed which just needed ultrasonic treatmen...A simple method to prepare of DNA template suitable for PCR amplification from filamentous fungi will be valuable for improving experimental efficiency.Here,a method was developed which just needed ultrasonic treatment of the mycelium at usual condition,and the produced solution could directly be used as DNA template for internal transcribed spacer(ITS)amplification successfully.The PCR could be improved by additional treatment of 60℃water baths,but was not centrifugation.When the template amount was 0.5-2μL and the ultrasonic time was 7-11 min,there was no distinctly influences on PCR.The method was commonly used for M.purpureus,I.cicadae,Lentinula sp.,Flammul sp.and Dictyophora sp.etc.to detect target sequences of ITS,hygromycin resistance gene(Hyg),CRISPR-associated protein 9(Cas9),Citrinin gene C(CtnC),Citrinin gene D(CtnD),large subunit rRNA gene(NL),and so on.The method could provide a simple,rapid,safe and economic approach to prepare the DNA template for large-scale PCR of the special filamentous fungi materials.展开更多
We have used the polymerase chain reaction to analyze variation in the size of 5S ribosomal gene spacer sequence. Eighty accessions, including 65 cultivated rice and 15 wild rice, were analyzed. Among them seven size ...We have used the polymerase chain reaction to analyze variation in the size of 5S ribosomal gene spacer sequence. Eighty accessions, including 65 cultivated rice and 15 wild rice, were analyzed. Among them seven size classes of 5S DNA spacer were observed. Classification asindica orjaponica on the basis of 5S DNA spacer patterns generally agrees with classification based on morphological studies, indicating that the length polymorphism of 5S DNA spacer could be used as a molecular marker for taxonomic and phylogenetic analysis.展开更多
The chromosomes 1, 3, 5, 6, 7,10 and 12 of rice field eel (Monopterus albus Zuiew) have been microdissected successfully from meiosis I diakinesis spreads by using glass microneedlc under an inverted microscope. And t...The chromosomes 1, 3, 5, 6, 7,10 and 12 of rice field eel (Monopterus albus Zuiew) have been microdissected successfully from meiosis I diakinesis spreads by using glass microneedlc under an inverted microscope. And the DOP-PCR products of the single chromosome dotted on the nylon membrane as 'specific chromosomal DNA pool', have been hybridized with 6 probes to map these genes. The mapping results show that Zfa has been mapped to chromosome 1, rDNA to chromosomes 3 and 7, both Gh and Pdegγto chromosome 10, Hsl to chromosome 5 and Hox genes have been detected on chromosomes 1, 3, 6 and 10 meantime. It has initiatively been suggested that chromosome 10 of rice field eel might possess the commom conserved synteny to that on chromosome 17 of human, chromosome 11 of mouse, chromosome 12 of pig and chromosome 19 of bovine. And so chromosome 3 of rice field eel might also contain the commom conserved synteny to that on chromosome 2 of zebraf-ish. Our study is an attempt to establish a new and feasible展开更多
基金Agricultural Science and Technology Innovation Fund of Hunan Province(2019LS06).
文摘[Objectives]To establish a simple,rapid and accurate method for identifying the genetic relationship of hybrid rice varieties to their parents.[Methods]Taking F1 hybrids Liangyou 336,Deliangyou Huazhan,and the parents of Liangyou 336,i.e.,C815S(♀)and R336(♂),as experimental materials,the genetic relationship of the hybrid rice varieties to the parental materials was identified by way of PCR amplification with the 48 pairs of SSR primers of Protocol for Identification of Rice Varieties:SSR Marker Method(NY/T 1433-2014).[Results]The genetic relationship of the hybrid rice varieties could be determined by comparing the PCR amplification products of the mixed DNA of the parents and the DNA of the F1 hybrids.[Conclusions]This method not only reduced the number of samples required but also had a good visual effect and high accuracy.
基金supported by grants from the Ministry of Agriculture of China for transgenic research(Grant No.2008ZX08009-003)the National Natural Science Foundation of China(Grant Nos.30710103903 and 30771160)the Natural Science Foundation of Zhejiang Province,China(Grant No.R3090023)
文摘The extraction of DNA is often the most time consuming and laborious step in high-throughput molecular genetic analysis and marker assisted selection （MAS） programs. A simple method for preparation of rice genomic DNA was developed. A small amount （1~50 mg） of leaf tissue of rice seedling, 500 pL of extraction buffer, and one steel bead were put into a 2-mL microcentrifuge tube. After vigorously mashing for 2 min, 5 μL of supernatant was directly applied to PCR amplification. Otherwise, the supematant was precipitated with two times volume of ethanol to obtain high quality genomic DNA. This method is simple, rapid, low cost, and reliable for PCR analysis. One person can manipulate as many as 96 samples for PCR in 10 min. It is especially suitable for genotyping of large number of samples.
基金Supported by the National Natural Science Foundation of China(81960692)the Science and Technology Support Program of Guizhou Province(2019-2776)。
文摘A simple method to prepare of DNA template suitable for PCR amplification from filamentous fungi will be valuable for improving experimental efficiency.Here,a method was developed which just needed ultrasonic treatment of the mycelium at usual condition,and the produced solution could directly be used as DNA template for internal transcribed spacer(ITS)amplification successfully.The PCR could be improved by additional treatment of 60℃water baths,but was not centrifugation.When the template amount was 0.5-2μL and the ultrasonic time was 7-11 min,there was no distinctly influences on PCR.The method was commonly used for M.purpureus,I.cicadae,Lentinula sp.,Flammul sp.and Dictyophora sp.etc.to detect target sequences of ITS,hygromycin resistance gene(Hyg),CRISPR-associated protein 9(Cas9),Citrinin gene C(CtnC),Citrinin gene D(CtnD),large subunit rRNA gene(NL),and so on.The method could provide a simple,rapid,safe and economic approach to prepare the DNA template for large-scale PCR of the special filamentous fungi materials.
文摘We have used the polymerase chain reaction to analyze variation in the size of 5S ribosomal gene spacer sequence. Eighty accessions, including 65 cultivated rice and 15 wild rice, were analyzed. Among them seven size classes of 5S DNA spacer were observed. Classification asindica orjaponica on the basis of 5S DNA spacer patterns generally agrees with classification based on morphological studies, indicating that the length polymorphism of 5S DNA spacer could be used as a molecular marker for taxonomic and phylogenetic analysis.
基金This work was supported by the National Natural Science Foundation of China (Grant No. 39970406)the Research Fund for the Doctoral Program of Higher Education of China (Grant No. 98048628).
文摘The chromosomes 1, 3, 5, 6, 7,10 and 12 of rice field eel (Monopterus albus Zuiew) have been microdissected successfully from meiosis I diakinesis spreads by using glass microneedlc under an inverted microscope. And the DOP-PCR products of the single chromosome dotted on the nylon membrane as 'specific chromosomal DNA pool', have been hybridized with 6 probes to map these genes. The mapping results show that Zfa has been mapped to chromosome 1, rDNA to chromosomes 3 and 7, both Gh and Pdegγto chromosome 10, Hsl to chromosome 5 and Hox genes have been detected on chromosomes 1, 3, 6 and 10 meantime. It has initiatively been suggested that chromosome 10 of rice field eel might possess the commom conserved synteny to that on chromosome 17 of human, chromosome 11 of mouse, chromosome 12 of pig and chromosome 19 of bovine. And so chromosome 3 of rice field eel might also contain the commom conserved synteny to that on chromosome 2 of zebraf-ish. Our study is an attempt to establish a new and feasible
