The crushing of leaves is the most time and effort-consuming part in DNA extraction which is a fundamental step in the study of molecular biology. In this study, a new rapid and batch-oriented crushing method for DNA ...The crushing of leaves is the most time and effort-consuming part in DNA extraction which is a fundamental step in the study of molecular biology. In this study, a new rapid and batch-oriented crushing method for DNA extraction from maize leaves was developed. In addition, the practicability of the developed method in molecular marker-assisted breeding was verified using SSR molecular maker technology so as to provide a rapid, batch-oriented, low-cost and non-toxic leafcrushing method for a large number of molecular marker tests, improving test efficiency.展开更多
RAPD （random amplified polymorphic DNA） markers were performed to fingerprint 20 varieties of maize. Twenty operon primers generated informative RAPD patterns and selected for further RAPD analysis. These 20 primers...RAPD （random amplified polymorphic DNA） markers were performed to fingerprint 20 varieties of maize. Twenty operon primers generated informative RAPD patterns and selected for further RAPD analysis. These 20 primers yielded 291 of main bands, out of which 169 were polymorphic bands across tested varieties. Each selected primer produced between 59 bands （OPC-01） to 142 bands （OPX-04）. Amplification products （DNA amplicons） and fragments size ranged from 260 bp （OPT-06） to 2,365 bp （OPP-05）. The largest number of polymorphic bands （20 bands） was produced by primer OPX-04 while, the lowest number of polymorphic bands （1 band） was produced by primer OPA-03. The primers of the most interest of this purpose were those that produced more variety specific DNA profiles, such as OPD-03, OPE-18, OPF-05, OPL-11 and OPX-04. The primer efficiency ranged from 0.20 in primer OPA-12 to 0.01 in primer OPA-03. The highest polymorphism and value of discrimination in this study were obtained with primers OPX-04, while, the lowest polymorphism and value of discrimination were obtained with primers OPA-03. The lowest genetic distance was 0.1941 between varieties Manlcet and Biotech Bag, while, the highest genetic distance was 0.6433 between varieties Pio 3751 and Buhooth 106. Cluster analysis （phylogenetic tree） by UPGMA （unweighted pair-group method of arithmetic means） based dendrogram revealed that they were four main genetic groups. The overall analysis of the results reveals that the genetic relationships among the maize varieties were related to some of their morphological characters as well as to their geographical origins at the molecular genetics.展开更多
The origin and evolution of B chromosomes could be explained by the specific DNA sequence on them. But the specific sequences known were quite limited. To investigate maize B chromosome sqicific DNA sequeces, maize ge...The origin and evolution of B chromosomes could be explained by the specific DNA sequence on them. But the specific sequences known were quite limited. To investigate maize B chromosome sqicific DNA sequeces, maize genomes with and without B chromosomes were analyzed by AFLP. Only 5 markers were found specific to genomes with B chromosomes among about 2000 AFLP markers. Southern hybridization and sequence analysis revealed that only the sequence of M8-2D was a B chromosome specific sequence. This sequence contained the telomeric repeat unit AGGGTTT conserved in plant chromosome telomeres. In addition, the sequence of M8-2D shared low homology to clones from maize chromosome 4 centromere as well. M8-2D were localized to B chromosome centrorneric and telomeric regions.展开更多
基金Supported by Special Fund for Agro-scientific Research in the Public Interest of China(201303008)Science and Technology Plan Project of Guangdong Province(2012B020301006)Key Breeding Project for Special Maize of Department of Agriculture of Guangdong Province(B3071328)~~
文摘The crushing of leaves is the most time and effort-consuming part in DNA extraction which is a fundamental step in the study of molecular biology. In this study, a new rapid and batch-oriented crushing method for DNA extraction from maize leaves was developed. In addition, the practicability of the developed method in molecular marker-assisted breeding was verified using SSR molecular maker technology so as to provide a rapid, batch-oriented, low-cost and non-toxic leafcrushing method for a large number of molecular marker tests, improving test efficiency.
文摘RAPD （random amplified polymorphic DNA） markers were performed to fingerprint 20 varieties of maize. Twenty operon primers generated informative RAPD patterns and selected for further RAPD analysis. These 20 primers yielded 291 of main bands, out of which 169 were polymorphic bands across tested varieties. Each selected primer produced between 59 bands （OPC-01） to 142 bands （OPX-04）. Amplification products （DNA amplicons） and fragments size ranged from 260 bp （OPT-06） to 2,365 bp （OPP-05）. The largest number of polymorphic bands （20 bands） was produced by primer OPX-04 while, the lowest number of polymorphic bands （1 band） was produced by primer OPA-03. The primers of the most interest of this purpose were those that produced more variety specific DNA profiles, such as OPD-03, OPE-18, OPF-05, OPL-11 and OPX-04. The primer efficiency ranged from 0.20 in primer OPA-12 to 0.01 in primer OPA-03. The highest polymorphism and value of discrimination in this study were obtained with primers OPX-04, while, the lowest polymorphism and value of discrimination were obtained with primers OPA-03. The lowest genetic distance was 0.1941 between varieties Manlcet and Biotech Bag, while, the highest genetic distance was 0.6433 between varieties Pio 3751 and Buhooth 106. Cluster analysis （phylogenetic tree） by UPGMA （unweighted pair-group method of arithmetic means） based dendrogram revealed that they were four main genetic groups. The overall analysis of the results reveals that the genetic relationships among the maize varieties were related to some of their morphological characters as well as to their geographical origins at the molecular genetics.
基金This research was supported by grant from National Natural Sciences Foundationof China (39970357).
文摘The origin and evolution of B chromosomes could be explained by the specific DNA sequence on them. But the specific sequences known were quite limited. To investigate maize B chromosome sqicific DNA sequeces, maize genomes with and without B chromosomes were analyzed by AFLP. Only 5 markers were found specific to genomes with B chromosomes among about 2000 AFLP markers. Southern hybridization and sequence analysis revealed that only the sequence of M8-2D was a B chromosome specific sequence. This sequence contained the telomeric repeat unit AGGGTTT conserved in plant chromosome telomeres. In addition, the sequence of M8-2D shared low homology to clones from maize chromosome 4 centromere as well. M8-2D were localized to B chromosome centrorneric and telomeric regions.
