A FISH procedure was adopted to physical mapping rice RFLP marker RZ69 and the BAC clone 38J9 screened by RZ69 which is linked to gene Bph3 in Oryza saliva L. and O. officinalis Well ex: Watt. The FISH results showed ...A FISH procedure was adopted to physical mapping rice RFLP marker RZ69 and the BAC clone 38J9 screened by RZ69 which is linked to gene Bph3 in Oryza saliva L. and O. officinalis Well ex: Watt. The FISH results showed that both 38J9 and RZ69 were located in the middle of 4S in O. officinalis and the centromere area of 4S in O. sativa. In O. officinalis the percentage distances from the centromere to the hybridization sites were 20.00 +/- 5.40 and 22.12 +/- 3.44, the detection rates were 50.00% and 6.14%, but in O. sativa they were 0 and 0, 56.10% and 6.29% correspondingly. The results obtained from the BAC and RFLP clone were almost the same in the cultivated rice and O. officinalis. It was suggested that die marker RZ69 of the cultivated rice and its homologous sequence in O. officinalis were in the same BAC clone, the homologous sequence of Bph3 in O. officinalis should be located at the sites hybridized by both RZ69 and 38J9. Many signals on different chromosomes of O. officinalis were observed under no blocking with Cot-1 DNA, showing that the repetitive sequences were also homologous between O. sativa and O. officinalis. The identification of chromosome 4 of O. officinalis is based on comparative map with RG214 and BAC clone screened by RG214. The feasibility of comparative physical mapping performed between O. sativa and O. officinalis with rice BAC clones was discussed.展开更多
Multiple BAC-FISH is a powerful tool for modern cytogenetic researching in both animals and plants.But in cotton,this technique is unavailable due to the high percentage of repetitive sequences.
The rice BAC-DNA was used as probes and fluorescence in situ hybridization(FISH)was applied to the interphase and metaphase mitotic chromosomes of maize.To optimize the BAC-FISH technique,we respectively assayed the e...The rice BAC-DNA was used as probes and fluorescence in situ hybridization(FISH)was applied to the interphase and metaphase mitotic chromosomes of maize.To optimize the BAC-FISH technique,we respectively assayed the effect of several factors,including maize or rice genomic C_(o)t DNA used as blocking reagent of DNA,washing temperatures and FAD concentration in the washing buffer and in the hybrid solution.The results show that C_(o)t DNA of maize genome blocked the repetitive sequence of the rice BAC-DNA when the C_(o)t value was below 50.Meanwhile,it was necessary to adjust the C_(o)t value according to the different probes and their ratios.Decreasing the concentration of FAD in the hybridization mixtures,adjusting the washing rate after hybridization,and most especially,blocking the ricespecific repetitive sequences of BAC-DNA could improve the positive signals of BAC-FISH.展开更多
A fluorescence in situ hybridization (FISH) procedure was adopted to physically map two rice BAC clones 24E21 and 4F22 linked to Gm-6 and Pi-5(t) in O. offi-cinalis. FISH results showed that the two BAC clones were lo...A fluorescence in situ hybridization (FISH) procedure was adopted to physically map two rice BAC clones 24E21 and 4F22 linked to Gm-6 and Pi-5(t) in O. offi-cinalis. FISH results showed that the two BAC clones were located at 4L. The percentage distance from the centromere to the hybridization sites was 72 ± 2.62 for 24E21 and 54± 5.43 for 4F22, the detection rates were 52.70% and 61.2%. The results obtained from the BAC and plasmid clones, RG214 and RZ565 of cultivated rice and O. officinalis were the same. This suggested that the markers, RG214 and RZ565 of cultivated rice and O. officinalis were in the same BAC clones. The homologous sequences of Gm-6 and Pi-5(t) in O. officinalis were positions that signals existed on the 4L. Many signals were observed when no Cot-1 DNA blocked. This also showed that repetitive sequences were some ho-molgous between cultivated rice and O. officinalis. The identification of chromosome 4 of O. officinalis is based on Jena et al. (1994). In our study, we discussed展开更多
Apomixis is a means of asexual reproduction by which plants produce embryos without fertilization and meiosis,therefore the embryo is of clonal and maternal origin.Interspecific hybrids between dip-loid B.vulgaris(2n=...Apomixis is a means of asexual reproduction by which plants produce embryos without fertilization and meiosis,therefore the embryo is of clonal and maternal origin.Interspecific hybrids between dip-loid B.vulgaris(2n=2x=18)and tetraploid B.corolliflora(2n=4x=36)were established,and then back-crossed with B.vulgaris.Among their offspring,monosomic addition line M14(2n=2x=18+1)was se-lected because of the apomictic phenotype.We documented chromosome transmission from B.corol-liflora into M14 by using genomic in situ hybridization(GISH).Suppression of cross-hybridization by blocking DNA was not necessary,indicating that the investigated Beta genome contains sufficient species-specific DNA,thus enabling the determination of genomic composition of the hybrids.We analyzed BAC microarrays of B.corolliflora chromosome 9 by using fluorescence-specific mRNA of B.vulgaris and Beta M14.BAC clones 16-M11 and 26-L15 were detected as fluorescence-specifics of BAC DNA of Beta M14.Then both BAC clones 16-M11 and 26-L15 were in situ hybridized to M14 chromo-somes.The two hybridized BAC clones were located close to the telomere region of the long arm of a single chromosome 9,and showed hemizygosity.The results of BAC microarrays showed that these developments of embryo and endosperm have conservative expression patterns,indicating that sexual reproduction and apomixis have an interrelated pathway with common regulatory components and that the induction of a modified sexual reproduction program may enable the manifestation of apomixis in Beta species.It would be sufficient for the expression of apomixes with those apomictic-specific genes on chromosome 9 of B.corolliflora.展开更多
文摘A FISH procedure was adopted to physical mapping rice RFLP marker RZ69 and the BAC clone 38J9 screened by RZ69 which is linked to gene Bph3 in Oryza saliva L. and O. officinalis Well ex: Watt. The FISH results showed that both 38J9 and RZ69 were located in the middle of 4S in O. officinalis and the centromere area of 4S in O. sativa. In O. officinalis the percentage distances from the centromere to the hybridization sites were 20.00 +/- 5.40 and 22.12 +/- 3.44, the detection rates were 50.00% and 6.14%, but in O. sativa they were 0 and 0, 56.10% and 6.29% correspondingly. The results obtained from the BAC and RFLP clone were almost the same in the cultivated rice and O. officinalis. It was suggested that die marker RZ69 of the cultivated rice and its homologous sequence in O. officinalis were in the same BAC clone, the homologous sequence of Bph3 in O. officinalis should be located at the sites hybridized by both RZ69 and 38J9. Many signals on different chromosomes of O. officinalis were observed under no blocking with Cot-1 DNA, showing that the repetitive sequences were also homologous between O. sativa and O. officinalis. The identification of chromosome 4 of O. officinalis is based on comparative map with RG214 and BAC clone screened by RG214. The feasibility of comparative physical mapping performed between O. sativa and O. officinalis with rice BAC clones was discussed.
文摘Multiple BAC-FISH is a powerful tool for modern cytogenetic researching in both animals and plants.But in cotton,this technique is unavailable due to the high percentage of repetitive sequences.
基金This research was supported by Major State Basic Research Development Program of China(No.2001CB108806).
文摘The rice BAC-DNA was used as probes and fluorescence in situ hybridization(FISH)was applied to the interphase and metaphase mitotic chromosomes of maize.To optimize the BAC-FISH technique,we respectively assayed the effect of several factors,including maize or rice genomic C_(o)t DNA used as blocking reagent of DNA,washing temperatures and FAD concentration in the washing buffer and in the hybrid solution.The results show that C_(o)t DNA of maize genome blocked the repetitive sequence of the rice BAC-DNA when the C_(o)t value was below 50.Meanwhile,it was necessary to adjust the C_(o)t value according to the different probes and their ratios.Decreasing the concentration of FAD in the hybridization mixtures,adjusting the washing rate after hybridization,and most especially,blocking the ricespecific repetitive sequences of BAC-DNA could improve the positive signals of BAC-FISH.
基金the National Natural Science Foundation ofChina (Grant No. 39870423) and the Doctorate Vesting Point Foundation of the State Education Commission of China (Grant No. 207980112).
文摘A fluorescence in situ hybridization (FISH) procedure was adopted to physically map two rice BAC clones 24E21 and 4F22 linked to Gm-6 and Pi-5(t) in O. offi-cinalis. FISH results showed that the two BAC clones were located at 4L. The percentage distance from the centromere to the hybridization sites was 72 ± 2.62 for 24E21 and 54± 5.43 for 4F22, the detection rates were 52.70% and 61.2%. The results obtained from the BAC and plasmid clones, RG214 and RZ565 of cultivated rice and O. officinalis were the same. This suggested that the markers, RG214 and RZ565 of cultivated rice and O. officinalis were in the same BAC clones. The homologous sequences of Gm-6 and Pi-5(t) in O. officinalis were positions that signals existed on the 4L. Many signals were observed when no Cot-1 DNA blocked. This also showed that repetitive sequences were some ho-molgous between cultivated rice and O. officinalis. The identification of chromosome 4 of O. officinalis is based on Jena et al. (1994). In our study, we discussed
基金the National Transgene Scientific Program (Grant No. JY03A2402)the Postdoctoral Science Foundation of Heilongjiang Province
文摘Apomixis is a means of asexual reproduction by which plants produce embryos without fertilization and meiosis,therefore the embryo is of clonal and maternal origin.Interspecific hybrids between dip-loid B.vulgaris(2n=2x=18)and tetraploid B.corolliflora(2n=4x=36)were established,and then back-crossed with B.vulgaris.Among their offspring,monosomic addition line M14(2n=2x=18+1)was se-lected because of the apomictic phenotype.We documented chromosome transmission from B.corol-liflora into M14 by using genomic in situ hybridization(GISH).Suppression of cross-hybridization by blocking DNA was not necessary,indicating that the investigated Beta genome contains sufficient species-specific DNA,thus enabling the determination of genomic composition of the hybrids.We analyzed BAC microarrays of B.corolliflora chromosome 9 by using fluorescence-specific mRNA of B.vulgaris and Beta M14.BAC clones 16-M11 and 26-L15 were detected as fluorescence-specifics of BAC DNA of Beta M14.Then both BAC clones 16-M11 and 26-L15 were in situ hybridized to M14 chromo-somes.The two hybridized BAC clones were located close to the telomere region of the long arm of a single chromosome 9,and showed hemizygosity.The results of BAC microarrays showed that these developments of embryo and endosperm have conservative expression patterns,indicating that sexual reproduction and apomixis have an interrelated pathway with common regulatory components and that the induction of a modified sexual reproduction program may enable the manifestation of apomixis in Beta species.It would be sufficient for the expression of apomixes with those apomictic-specific genes on chromosome 9 of B.corolliflora.
