In this study,silkworm strain T6,tolerant to fluoride,and silkworm strain 733xin,highly sensitive to fluoride,were used to construct the near-isogenic lines.300 random primers were used in RAPD amplification to DNAs o...In this study,silkworm strain T6,tolerant to fluoride,and silkworm strain 733xin,highly sensitive to fluoride,were used to construct the near-isogenic lines.300 random primers were used in RAPD amplification to DNAs of these lines.A molecular marker named S207 was found linked to the fluoride tolerance gene.Examination to F 2 segregated individuals of the above lines verified that this molecular marker was reliable.Subsequently,the molecular marker was cloned into a T vector (pUCm-T) for sequencing.Comparing with sequences available in the GenBank showed that this molecular marker was novel.We plan to convert it into a SCAR marker to facilitate establishment of a molecular marker assisted breeding system.展开更多
AFLP analysis was performed between a pair of thermo_sensitive genic male sterile (TGMS) rice allelic mutant lines (5460S and 5460F). The reaction conditions for rice AFLP assay were optimized. The relative efficienci...AFLP analysis was performed between a pair of thermo_sensitive genic male sterile (TGMS) rice allelic mutant lines (5460S and 5460F). The reaction conditions for rice AFLP assay were optimized. The relative efficiencies for polymorphism detection of RFLP, RAPD and AFLP were compared. The results indicated that the efficiency for polymorphism detection in rice was in the order of AFLP>RAPD>RFLP, and also indicated that AFLP was a powerful DNA molecular marker technique for polymorphism detection, especially in the case of extremely low polymorphism, such as isogenic lines and allelic mutant lines. Some of the AFLP products between the TGMS rice allelic mutant lines were cloned. Three of them were used as mixed probes to screen BAC library of rice line 5460S. 12 positive clones were screened out. In addition, the advantages and disadvantages of these three molecular marker systems were discussed.展开更多
Cassava mosaic disease （CMD）, the most important disease of cassava （Manihot esculenta Crantz） is a potential threat to Africa＇s cassava production. The disease is embedded in most landraces resulting in low yiel...Cassava mosaic disease （CMD）, the most important disease of cassava （Manihot esculenta Crantz） is a potential threat to Africa＇s cassava production. The disease is embedded in most landraces resulting in low yields. Host plant resistance has been found to be the best control strategy. A breeding programme using genetic hybridisation and Marker Assisted Selection was initiated in 2007 to improve the resistance levels of farmer-preferred landraces and reduce the long breeding cycle for developing improved cassava varieties. Thirty farmer-preferred landraces were selected and crossed with a high yielding and mosaic resistant cultivar （TMEI l） from International Institute of Tropical Agriculture （IITA）. The resultant progenies found to have a reasonable level of resistance （score, 1-3） to the CMD were AW 18, 273 Nyamebekyere, NK 43, AW 3, NK 26, K 25, Dabodabo, Ahwengyankwa, 674 Debor, Degarti, Agric Bankye, and NK 57. These resultant progenies were backcrossed to the resistant cultivar. The backcross one （BC l） progenies totaling 224 were screened with molecular markers that are associated to the CMD 2 gene. De-oxyribonucleic Acid （DNA） was extracted from leaves of the 224 BC1 progenies and 13 parents. Two Simple Sequence Repeats （SSR） markers （SSY28 and NSl58） and one Sequenced Characterized Amplified Region （SCAR） marker RME1, were used to screen and select for the resistant BC l progenies. Marker Assisted Selection （MAS） revealed that 82% of the genotypes had at least a marker allele for the CMD2 gene, indicating resistance. The study further revealed that by using MAS, the breeding cycle of cassava in the generation of varieties could be reduced from 8 years to 2 years. The resistant genotypes identified will be evaluated for yield and starch cooking quality in future breeding work.展开更多
文摘In this study,silkworm strain T6,tolerant to fluoride,and silkworm strain 733xin,highly sensitive to fluoride,were used to construct the near-isogenic lines.300 random primers were used in RAPD amplification to DNAs of these lines.A molecular marker named S207 was found linked to the fluoride tolerance gene.Examination to F 2 segregated individuals of the above lines verified that this molecular marker was reliable.Subsequently,the molecular marker was cloned into a T vector (pUCm-T) for sequencing.Comparing with sequences available in the GenBank showed that this molecular marker was novel.We plan to convert it into a SCAR marker to facilitate establishment of a molecular marker assisted breeding system.
文摘AFLP analysis was performed between a pair of thermo_sensitive genic male sterile (TGMS) rice allelic mutant lines (5460S and 5460F). The reaction conditions for rice AFLP assay were optimized. The relative efficiencies for polymorphism detection of RFLP, RAPD and AFLP were compared. The results indicated that the efficiency for polymorphism detection in rice was in the order of AFLP>RAPD>RFLP, and also indicated that AFLP was a powerful DNA molecular marker technique for polymorphism detection, especially in the case of extremely low polymorphism, such as isogenic lines and allelic mutant lines. Some of the AFLP products between the TGMS rice allelic mutant lines were cloned. Three of them were used as mixed probes to screen BAC library of rice line 5460S. 12 positive clones were screened out. In addition, the advantages and disadvantages of these three molecular marker systems were discussed.
文摘Cassava mosaic disease （CMD）, the most important disease of cassava （Manihot esculenta Crantz） is a potential threat to Africa＇s cassava production. The disease is embedded in most landraces resulting in low yields. Host plant resistance has been found to be the best control strategy. A breeding programme using genetic hybridisation and Marker Assisted Selection was initiated in 2007 to improve the resistance levels of farmer-preferred landraces and reduce the long breeding cycle for developing improved cassava varieties. Thirty farmer-preferred landraces were selected and crossed with a high yielding and mosaic resistant cultivar （TMEI l） from International Institute of Tropical Agriculture （IITA）. The resultant progenies found to have a reasonable level of resistance （score, 1-3） to the CMD were AW 18, 273 Nyamebekyere, NK 43, AW 3, NK 26, K 25, Dabodabo, Ahwengyankwa, 674 Debor, Degarti, Agric Bankye, and NK 57. These resultant progenies were backcrossed to the resistant cultivar. The backcross one （BC l） progenies totaling 224 were screened with molecular markers that are associated to the CMD 2 gene. De-oxyribonucleic Acid （DNA） was extracted from leaves of the 224 BC1 progenies and 13 parents. Two Simple Sequence Repeats （SSR） markers （SSY28 and NSl58） and one Sequenced Characterized Amplified Region （SCAR） marker RME1, were used to screen and select for the resistant BC l progenies. Marker Assisted Selection （MAS） revealed that 82% of the genotypes had at least a marker allele for the CMD2 gene, indicating resistance. The study further revealed that by using MAS, the breeding cycle of cassava in the generation of varieties could be reduced from 8 years to 2 years. The resistant genotypes identified will be evaluated for yield and starch cooking quality in future breeding work.
