A simple and efficient method was presented for isolating microsatellite DNA markers from peanut (Arachis hypogaea L.) genome. The genomic DNA was converted into pre-amplified AFLP fragments and hybridized with biotin...A simple and efficient method was presented for isolating microsatellite DNA markers from peanut (Arachis hypogaea L.) genome. The genomic DNA was converted into pre-amplified AFLP fragments and hybridized with biotin-labeled SSR probes. Then the hybrid mixture was used to incubate with magnetic beads coated with streptavidin. After washing to remove the non-SSR fragments, the eluted single-strand DNA, which was cloned and sequenced, was largely enriched for microsatellites. Primers can then be designed according to the sequence flanking the repeat motifs and used for polymorphism analysis. The whole experiment can be completed within one week and can be employed as a reliable option for any molecular laboratory to develop SSR markers.展开更多
In order to screen molecular markers linked to fertility restoring genes and further improve the breeding efficiency of restorer lines, in this study, wheat varieties 18A, 18B and 99AR144-1 were used as experimental m...In order to screen molecular markers linked to fertility restoring genes and further improve the breeding efficiency of restorer lines, in this study, wheat varieties 18A, 18B and 99AR144-1 were used as experimental materials to establish F2 fertility-segregating population. Plant quantitative trait ＂major gene ＋ polygene mixed mo- del＂ separation analysis method and simple sequence repeat （SSR） molecular markers were adopted for genetic analysis of four generations, including the parents （P~ and P2）, and hybrid （G and G） populations. The results show that AL-type fertility restoring gene is controlled by two pairs of additive-dominant-epistatic genes and addi- tive-dominant polygene; two primers linked to fertility restoring genes were selected by SSR molecular markers, including Xgwm95 on chromosome 2A and Barc61 on chromosome 1B, with the linkage distance of 15.0 cM and 18.0 cM, respectively. Based on verification, these two markers are reliable for distinguishing AL-type wheat ste- rile lines and restorer lines.展开更多
A RIL population was used in this study, which was derived from a cross between a temperate maize germplasm inbred line B31-1 and a tropical maize germplasm inbred lines Huangzao 4. A genetic linkage map was construct...A RIL population was used in this study, which was derived from a cross between a temperate maize germplasm inbred line B31-1 and a tropical maize germplasm inbred lines Huangzao 4. A genetic linkage map was constructed comprising of 153 polymorphic markers. Among the 153 polymorphic markers, 82 markers showed the significantly segregation distortion（P〈0.05）, favoring either the marker alleles of female parent 1331-1（62.50%） or male parent Huangzao 4（37.50%）. Segregation distortion marker distribution along the present molecular maps of maize was far from uniform, with clusters of tightly linked loci and single marker. Nine segregation distortion regions were detected on 10 chromosomes, indicating that possible causes for segregation deviation of molecular markers are genetic selection.展开更多
Microsatellite markers have become one kind of the most important molecular tools used in various researches. A large number of microsatellite markers are required for the whole genome survey in the fields of molecula...Microsatellite markers have become one kind of the most important molecular tools used in various researches. A large number of microsatellite markers are required for the whole genome survey in the fields of molecular ecology,quantitative genetics and genomics. Therefore,it is extremely necessary to select several versatile,low-cost,efficient and time-and labor-saving methods to develop a large panel of microsatellite markers. In this study,we used Zhikong scallop(Chlamys farreri) as the target species to compare the efficiency of the five methods derived from three strategies for microsatellite marker development. The results showed that the strategy of constructing small insert genomic DNA library resulted in poor efficiency,while the microsatellite-enriched strategy highly improved the isolation efficiency. Although the mining public database strategy is time-and cost-saving,it is difficult to obtain a large number of microsatellite markers,mainly due to the limited sequence data of non-model species deposited in public databases. Based on the results in this study,we recommend two methods,microsatellite-enriched library construction method and FIASCO-colony hybridization method,for large-scale microsatellite marker development. Both methods were derived from the microsatellite-enriched strategy. The experimental results obtained from Zhikong scallop also provide the reference for microsatellite marker development in other species with large genomes.展开更多
Two silkworm strains viz, B20 A (high cocoon shell ratio) and C.Nichi (low cocoon shell ratio) were sib mated for 10 generations to determine the homozygosis. Both bulked segregant analysis(BSA) and near isogenic line...Two silkworm strains viz, B20 A (high cocoon shell ratio) and C.Nichi (low cocoon shell ratio) were sib mated for 10 generations to determine the homozygosis. Both bulked segregant analysis(BSA) and near isogenic lines (NIL) studies were done to identify the RFLP markers closely linked to cocoon shell parameters. Three hundred and fifty two random clones were identified as the low copy number sequence and used for identification of Restriction Fragment Length Polymorphic (RFLP) marker linked to cocoon weight and cocoon shell character. In the bulk segregant analysis, DNA from the parents (B20 A, C.Nichi), F 1 and F 2 progeny of high shell ratio (HSR) and low shell ratio (LSR) were screened for hybridization with the random clones. Polymorphic banding pattern achieved through southern hybridization with different probes indicated the probable correlation of polymorphism with high and low cocoon shell character which are possible landmarks in identifying the putative marker(s) for the cocoon shell character. Out of the 100 probes tried with parents, F 1, F 2 and their bulks, 10 probes were found to be closely linked to cocoon shell characters.展开更多
目的建立22q11微缺失综合征的产前诊断方法。方法应用细菌人工染色体标记一磁珠鉴别／分离技术（BACs-on-Beads^TM，BoBs）和荧光原位杂交技术（fluorescence in situ hybridization，FISH），对1例羊水染色体培养失败的胎儿及1例疑似22...目的建立22q11微缺失综合征的产前诊断方法。方法应用细菌人工染色体标记一磁珠鉴别／分离技术（BACs-on-Beads^TM，BoBs）和荧光原位杂交技术（fluorescence in situ hybridization，FISH），对1例羊水染色体培养失败的胎儿及1例疑似22q11微缺失综合征的双胎行产前分子诊断。结果产前BoBs试剂盒方法检测到1例胎儿及1例双胎均为22q11的微缺失，同时3例胎儿中期分裂细胞的FISH验证结果均显示为22q11微缺失，即在DiGeorge／VCFSN25位点上仅有一个红色荧光信号，对照22号末端22q13．3ARSA位点上有两个绿色荧光信号。结论产前Bobs方法联合FISH技术可以成为22q11微缺失的一种产前分子诊断手段。展开更多
文摘A simple and efficient method was presented for isolating microsatellite DNA markers from peanut (Arachis hypogaea L.) genome. The genomic DNA was converted into pre-amplified AFLP fragments and hybridized with biotin-labeled SSR probes. Then the hybrid mixture was used to incubate with magnetic beads coated with streptavidin. After washing to remove the non-SSR fragments, the eluted single-strand DNA, which was cloned and sequenced, was largely enriched for microsatellites. Primers can then be designed according to the sequence flanking the repeat motifs and used for polymorphism analysis. The whole experiment can be completed within one week and can be employed as a reliable option for any molecular laboratory to develop SSR markers.
基金Special Foundation for "12th Five-year" Biological Germplasm Resources Innovation&Functional Gene Discovery and Utilization of Xinjiang Production and Construction Corps(No.2012BB047)"12th Five-year" Breeding Tacking Program of Xinjiang Production and Construction Corps(No.2011BA002)
文摘In order to screen molecular markers linked to fertility restoring genes and further improve the breeding efficiency of restorer lines, in this study, wheat varieties 18A, 18B and 99AR144-1 were used as experimental materials to establish F2 fertility-segregating population. Plant quantitative trait ＂major gene ＋ polygene mixed mo- del＂ separation analysis method and simple sequence repeat （SSR） molecular markers were adopted for genetic analysis of four generations, including the parents （P~ and P2）, and hybrid （G and G） populations. The results show that AL-type fertility restoring gene is controlled by two pairs of additive-dominant-epistatic genes and addi- tive-dominant polygene; two primers linked to fertility restoring genes were selected by SSR molecular markers, including Xgwm95 on chromosome 2A and Barc61 on chromosome 1B, with the linkage distance of 15.0 cM and 18.0 cM, respectively. Based on verification, these two markers are reliable for distinguishing AL-type wheat ste- rile lines and restorer lines.
基金Supported by the Fund for Breeding and Commercial Seed Production of Functional Maize Materials(cstc2016shms-ztzx80013)the Fund for Commercial Seed Production Innovation of New Silage Maize Varieties(cstc2016shms-ztzx80015)+1 种基金the Fund for Breeding and Mechanized Production of High-quality and High-Yield Silage Maize Varieties in Southwest China(2016YFD0300309-4)Chongqing basic scientific research fund in 2016(Discovery and Application of Key Gene for Selenium Accumulation in Maize Grains)~~
文摘A RIL population was used in this study, which was derived from a cross between a temperate maize germplasm inbred line B31-1 and a tropical maize germplasm inbred lines Huangzao 4. A genetic linkage map was constructed comprising of 153 polymorphic markers. Among the 153 polymorphic markers, 82 markers showed the significantly segregation distortion（P〈0.05）, favoring either the marker alleles of female parent 1331-1（62.50%） or male parent Huangzao 4（37.50%）. Segregation distortion marker distribution along the present molecular maps of maize was far from uniform, with clusters of tightly linked loci and single marker. Nine segregation distortion regions were detected on 10 chromosomes, indicating that possible causes for segregation deviation of molecular markers are genetic selection.
基金supported by ‘863’ Program (2006AA10A408 and 2006AA10A411), NSFC30571417, NYHYZX07-047, 2005DKA30470, 2006BAD09A10 and NCET-06-0594.
文摘Microsatellite markers have become one kind of the most important molecular tools used in various researches. A large number of microsatellite markers are required for the whole genome survey in the fields of molecular ecology,quantitative genetics and genomics. Therefore,it is extremely necessary to select several versatile,low-cost,efficient and time-and labor-saving methods to develop a large panel of microsatellite markers. In this study,we used Zhikong scallop(Chlamys farreri) as the target species to compare the efficiency of the five methods derived from three strategies for microsatellite marker development. The results showed that the strategy of constructing small insert genomic DNA library resulted in poor efficiency,while the microsatellite-enriched strategy highly improved the isolation efficiency. Although the mining public database strategy is time-and cost-saving,it is difficult to obtain a large number of microsatellite markers,mainly due to the limited sequence data of non-model species deposited in public databases. Based on the results in this study,we recommend two methods,microsatellite-enriched library construction method and FIASCO-colony hybridization method,for large-scale microsatellite marker development. Both methods were derived from the microsatellite-enriched strategy. The experimental results obtained from Zhikong scallop also provide the reference for microsatellite marker development in other species with large genomes.
文摘Two silkworm strains viz, B20 A (high cocoon shell ratio) and C.Nichi (low cocoon shell ratio) were sib mated for 10 generations to determine the homozygosis. Both bulked segregant analysis(BSA) and near isogenic lines (NIL) studies were done to identify the RFLP markers closely linked to cocoon shell parameters. Three hundred and fifty two random clones were identified as the low copy number sequence and used for identification of Restriction Fragment Length Polymorphic (RFLP) marker linked to cocoon weight and cocoon shell character. In the bulk segregant analysis, DNA from the parents (B20 A, C.Nichi), F 1 and F 2 progeny of high shell ratio (HSR) and low shell ratio (LSR) were screened for hybridization with the random clones. Polymorphic banding pattern achieved through southern hybridization with different probes indicated the probable correlation of polymorphism with high and low cocoon shell character which are possible landmarks in identifying the putative marker(s) for the cocoon shell character. Out of the 100 probes tried with parents, F 1, F 2 and their bulks, 10 probes were found to be closely linked to cocoon shell characters.
文摘目的建立22q11微缺失综合征的产前诊断方法。方法应用细菌人工染色体标记一磁珠鉴别／分离技术（BACs-on-Beads^TM，BoBs）和荧光原位杂交技术（fluorescence in situ hybridization，FISH），对1例羊水染色体培养失败的胎儿及1例疑似22q11微缺失综合征的双胎行产前分子诊断。结果产前BoBs试剂盒方法检测到1例胎儿及1例双胎均为22q11的微缺失，同时3例胎儿中期分裂细胞的FISH验证结果均显示为22q11微缺失，即在DiGeorge／VCFSN25位点上仅有一个红色荧光信号，对照22号末端22q13．3ARSA位点上有两个绿色荧光信号。结论产前Bobs方法联合FISH技术可以成为22q11微缺失的一种产前分子诊断手段。
文摘目的产前诊断1例超声异常的Miller-Dieker（Miller—Dieker syndrome，MDS）胎儿，分析并探讨其基因型与表型的对应关系。方法联合应用染色体核型分析、细菌人工染色体标记一磁珠鉴别／分离技术（BACs—on-Beads，BoBs）、荧光原位杂交技术（fluorescence in situ hybridization，FISH）和单核苷酸多态性微阵列技术对1例超声异常的胎儿进行产前诊断。结果产前BoBs检测提示胎儿携带17p13区的MDS微缺失，中期分裂细胞FISH确认其为17p13区的微缺失，高分辨的单核苷酸多态微阵列检测确定该胎儿染色体在17p13区存在约5．2Mb的缺失：arr[hg19]17p13．3p13．2（525—5204373）×1。胎儿脐血细胞染色体核型分析、孕妇及其配偶的外周血高分辨染色体核型分析和BoBs检测均未见异常。结论联合应用产前分子遗传学技术对1例新发的MDS综合征胎儿进行了产前诊断，临床上应重视微缺失微重复的病例，避免漏诊。
