Single pollen grain polymerase chain reaction （PCR） has succeeded in several species, however only limited numbers of pollen grains were involved due to difficulties in pollen isolation and lysis. This has limited i...Single pollen grain polymerase chain reaction （PCR） has succeeded in several species, however only limited numbers of pollen grains were involved due to difficulties in pollen isolation and lysis. This has limited its application in genetic analysis and mapping studies in plants. A high-throughput （HT） procedure for collecting and detecting genetic variation in a large number of individual pollen grains by PCR is reported. The HT procedure involved the collection of individual pollen grains by a pair of special forceps and the lysis of pollen grains in a heated alkali/detergent solution followed by neutralization with a tris-ethylenediamine tetraacetic acid （TE） buffer. These resulting template solutions yielded PCR reactions involving the 5S ribosomal RNA intergenic spacers, randomly amplified polymorphic DNA, and simple sequence repeats markers. Using this procedure, one person with experience could collect and process up to 288 single pollen grain PCR reactions per day. The method worked well on sugarcane, corn, Miscanthus spp., snap bean, sorghum, and tomato. The ability to collect and conduct PCR on individual pollen grains on a large scale offers a new approach to genetic analyses and mapping studies in an easily controllable environment with a considerable cost reduction. The method will also significantly benefit studies in species that are difficult subjects for classical genetic research.展开更多
Electrophoresis and staining of proteins from the single pollen grains of Hibiscus rosa sinensis have been developed by using general ultrathin polyacrylamide gel combined with highly sensitive silver staining techniq...Electrophoresis and staining of proteins from the single pollen grains of Hibiscus rosa sinensis have been developed by using general ultrathin polyacrylamide gel combined with highly sensitive silver staining technique.The result revealed that the pollen abortion could occur in different stages of pollen development.The protein patterns varied greatly in different stages of pollen development, even in the different pollen grains in the same anther at the same development stage.Some bands exhibited a disjunction by classical Mendelian ratio 1∶1, suggesting that the gene loci were heterogeneous and the proteins were related to the expression of the genes at the early stage of pollen development.展开更多
基金Grower/processor Check-off Funds administrated by theAmerican Sugar Cane League of the USA., Inc., Thibodaux, Louisiana, USA(to Y.-B. Pan)the Chinese 948 Project (2003-Q06) (to P.-H. Chen).
文摘Single pollen grain polymerase chain reaction （PCR） has succeeded in several species, however only limited numbers of pollen grains were involved due to difficulties in pollen isolation and lysis. This has limited its application in genetic analysis and mapping studies in plants. A high-throughput （HT） procedure for collecting and detecting genetic variation in a large number of individual pollen grains by PCR is reported. The HT procedure involved the collection of individual pollen grains by a pair of special forceps and the lysis of pollen grains in a heated alkali/detergent solution followed by neutralization with a tris-ethylenediamine tetraacetic acid （TE） buffer. These resulting template solutions yielded PCR reactions involving the 5S ribosomal RNA intergenic spacers, randomly amplified polymorphic DNA, and simple sequence repeats markers. Using this procedure, one person with experience could collect and process up to 288 single pollen grain PCR reactions per day. The method worked well on sugarcane, corn, Miscanthus spp., snap bean, sorghum, and tomato. The ability to collect and conduct PCR on individual pollen grains on a large scale offers a new approach to genetic analyses and mapping studies in an easily controllable environment with a considerable cost reduction. The method will also significantly benefit studies in species that are difficult subjects for classical genetic research.
文摘Electrophoresis and staining of proteins from the single pollen grains of Hibiscus rosa sinensis have been developed by using general ultrathin polyacrylamide gel combined with highly sensitive silver staining technique.The result revealed that the pollen abortion could occur in different stages of pollen development.The protein patterns varied greatly in different stages of pollen development, even in the different pollen grains in the same anther at the same development stage.Some bands exhibited a disjunction by classical Mendelian ratio 1∶1, suggesting that the gene loci were heterogeneous and the proteins were related to the expression of the genes at the early stage of pollen development.
