Longiflorum and Asiatic lilies of the genus Lilium of the family Liliaceae are two important groups of modem lily cultivars. One of the main trends of lily breeding is to realize introgression between these groups. Wi...Longiflorum and Asiatic lilies of the genus Lilium of the family Liliaceae are two important groups of modem lily cultivars. One of the main trends of lily breeding is to realize introgression between these groups. With cut style pollination and embryo rescue, distant hybrids between the two groups have been obtained. However, the FI hybrids are highly sterile or some of them could produce a small number of 2n gametes, and their BC1 progenies are usually triploids. Dutch lily breeders have selected many cultivars from these BC1 progenies based on their variation. It is presumably suggested that such variation could be caused by intergenomic recombination and abnormal meiosis during gamete formation in F1 hybrids of Longiflorum × Asiatic (LA) hybrids in Lilium. Therefore, the meiotic process of ten F1 LA hybrids was cytologically investigated using genomic in situ hybridization and traditional cytological methods in the present research. The results showed that: at metaphase I, the homoeologous chromosome pairing among different F1 hybrids ranged from 2.0 to 11.4 bivalents formed by homoeologous chromosomes per pollen mother cell (PMC), and very few multivalents, and even very few bivalents were formed by two chromosomes within one genome rather than homoeologous chromosomes in some PMCs; at anaphase I, all biva- lents were disjoined and most univalents were divided. Both the disjoined bivalents (half-bivalents) and the divided univalents (sister chromatids) moved to the opposite poles, and then formed two groups of chromosomes; because the two resulting half-bivalents retained their axes in the cell undisturbed, many crossover types, including single crossovers, three strand double crossovers, four strand double crossovers, four strand triple crossovers, and four strand multiple crossovers between the non-sister chromatids in the tetrads of bivalents, were clearly inferred by analyzing the breakpoints on the disjoined bivalents. The present investigation not only explained the reason for sterility of the Fl LA hybrids and the variation of their BCx progenies, but also provided a new method to analyze crossover types in other F1 interspecific hybrids as well.展开更多
To transfer desirable resistance traits from O. minuta to O. sativa, intergeneric hybrid plants between O. sativa (AA, 2n=2X=24) and O. minuta (BBCC, 2n=4X=48) were produced by embryo rescue after sexual cross. Morpho...To transfer desirable resistance traits from O. minuta to O. sativa, intergeneric hybrid plants between O. sativa (AA, 2n=2X=24) and O. minuta (BBCC, 2n=4X=48) were produced by embryo rescue after sexual cross. Morphological observation and chromosome counts indicated their hybrid status (ABC, 2n=3X=36). Genomic in situ hybridization (GISH) was further applied to confirm the parentage of the chromosomes of F 1 hybrids. Chromosomes of O. minuta and O. sativa were distinguishable in the hybrids in different fluorescence colors. GISH indicated that A and BC chromosomes were not randomly assembled in a cell. RAPD profiles unequivocally revealed their hybrids with double parent patterns. The results of blast tests showed that the hybrids had obtained disease resistance from O. minuta, and had a level of susceptibility between the parents.展开更多
In the genus Oryza, interspecific hybrids are useful bridges for transferring the desired genes from wild species to cultivated rice (Oryza sativa L.). In the present study, hybrids between O. sativa (AA genome) a...In the genus Oryza, interspecific hybrids are useful bridges for transferring the desired genes from wild species to cultivated rice (Oryza sativa L.). In the present study, hybrids between O. sativa (AA genome) and three Chinese wild rices, namely O. rufipogon (AA genome), O. officinalis (CC genome), and O. meyeriana (GG genome), were produced. Agricultural traits of the F1 hybrids surveyed were intermediate between their parents and appreciably resembled wild rice parents. Except for the O. sativa × O. rufipogon hybrid, the other F1 hybrids were completely sterile. Genomic in situ hybridization (GISH) was used for hybrid verification. Wild rice genomic DNAs were used as probes and cultivated rice DNA was used as a block. With the exception of O. rufipogon chromosomes, this method distinguished the other two wild rice and cultivated rice chromosomes at the stage of mitotic metaphase with different blocking ratios. The results suggest that a more distant phylogenetic relationship exists between O. meyeriana and O. sativa and that O. rufipogon and O. sativa share a high degree of sequence homology. The average mitotic chromosome length of O. officinalis and O. meyeriana was 1.25- and 1.51-fold that of O. sativa, respectively. 4',6'-Diamidino- 2-phenylindole staining showed that the chromosomes of O. officinalis and O. meyeriana harbored more heterochromatin, suggesting that the C and G genomes were amplified with repetitive sequences compared with the A genome. Although chromocenters formed by chromatin compaction were detected with wild rice-specific signals corresponding to the C and G genomes in discrete domains of the F1 hybrid interphase nuclei, the size and number of O. meyeriana chromocenters were bigger and greater than those of O. officinalis. The present results provide an important understanding of the genomic relationships and a tool for the transfer of useful genes from three native wild rice species in China to cultivars.展开更多
Gossypium mustelinum [(AD)4] is one of five tetraploid species in Gossypium.Three pairs of nucleolar organizer regions(NOR) in(AD)4 were detected by FISH with 45S rDNA as a probe,they also were observed with genomic D...Gossypium mustelinum [(AD)4] is one of five tetraploid species in Gossypium.Three pairs of nucleolar organizer regions(NOR) in(AD)4 were detected by FISH with 45S rDNA as a probe,they also were observed with genomic DNA(gDNA) from Gossypium D genome species as probes.Of展开更多
The St and E are two important basic genomes in the perennial tribe Triticeae (Poaceae). They exist in many perennial species and are very closely related to the A, B and D genomes of bread wheat (Triticum aestivum...The St and E are two important basic genomes in the perennial tribe Triticeae (Poaceae). They exist in many perennial species and are very closely related to the A, B and D genomes of bread wheat (Triticum aestivum L.). Genomic Southern hybridization and genomic in situ hybridization (GISH) were used to analyze the genomic relationships between the two genomes (St and E) and the three basic genomes (A, B and D) of T. aestivum. The semi-quantitative analysis of the Southern hybridization suggested that both St and E genomes are most closely related to the D genome, then the A genome, and relatively distant to the B genome. GISH analysis using St and E genomic DNA as probes further confirmed the conclusion. St and E are the two basic genomes of Thinopyrum ponticum (StStE^eE^bE^x) and Th. intermedium (StE^eE^b), two perennial species successfully used in wheat improvement. Therefore, this paper provides a possible answer as to why most of the spontaneous wheat-Thinopyrum translocations and substitutions usually happen in the D genome, some in the A genome and rarely in the B genome. This would develop further use of alien species for wheat improvement, especially those containing St or E in their genome components.展开更多
Using the genomic DNAs of maize and rice as probes respectively, the homology of maize and rice genomes was assessed by genomic in situ hybridization. When rice genomic DNAs were hybridized to maize, all chromosomes d...Using the genomic DNAs of maize and rice as probes respectively, the homology of maize and rice genomes was assessed by genomic in situ hybridization. When rice genomic DNAs were hybridized to maize, all chromosomes displayed many multiple discrete regions, while each rice chromosome delineated a single consecutive chromosomal region after they were hybridized with maize genomic DNAs. The results indicate that the genomes of maize and rice share high homology, and confirm the proposal that maize and rice are diverged from a common ancestor.展开更多
To identify alien chromosomes in recipient progenies and to analyze genome components in polyploidy, a genomic in situ hybridization (GISH) technique that is suitable for cotton was developed using increased stringe...To identify alien chromosomes in recipient progenies and to analyze genome components in polyploidy, a genomic in situ hybridization (GISH) technique that is suitable for cotton was developed using increased stringency conditions. The increased stringency conditions were a combination of the four factors in the following optimized state: 100:1 ratio of blocking DNA to probe, 60% formamide wash solution, 43 ℃ temperature wash and a 13 min wash. Under these specific conditions using gDNA from Gossypium sturtianum (C1 C1 ) as a probe, strong hybridization signals were only observed on chromosomes from the C1 genome in somatic cells of the hybrid F1 (G. hirsutum x G. sturtianum) (AtDtC1). Therefore, GISH was able to discriminate parental chromosomes in the hybrid. Further, we developed a multi-color GISH to simultaneously discriminate the three genomes of the above hybrid. The results repeatedly displayed the three genomes, At, Dt, and C1, and each set of chromosomes with a unique color, making them easy to identify. The power of the multi-color GISH was proven by analysis of the hexaploid hybrid F1 (G. hirsutum x G. australe) (AtAtDtDtG2G2). We believe that the powerful multi-color GISH technique could be applied extensively to analyze the genome component in polyploidy and to identify alien chromosomes in the recipient progenies.展开更多
Genomic in situ hybridization banding (GISH-banding), a technique slightly modified from conventional GISH, was used to probe the Chinese native rye (Secale cereale L.) DNA, and enabled us to visualize the individ...Genomic in situ hybridization banding (GISH-banding), a technique slightly modified from conventional GISH, was used to probe the Chinese native rye (Secale cereale L.) DNA, and enabled us to visualize the individual rye chromosomes and create a universal reference karyotype of the S. cereale chromosome 1R to 7R. The GISH-banding approach used in the present study was able to discriminate S. cereale chromosomes or segments in the wheat (Triticum aestivum L.) background, including the Triticale, wheat-rye addition and translocation lines. Moreover, the GISH-banding pattern of S. cereale subsp. Afghanicum chromosomes was consistent with that of Chinese native rye cv. Jingzhou rye; whereas the GISH-banding pattern of Secale vavilovii was different from that of S. cereale, indicating that GISH-banding can be used to study evolutionary polymorphism in species or subspecies of Secale. In addition, the production and application of GISH-banding to the study of adenine-thymine-riched heterochromaUn is discussed.展开更多
以栉孔扇贝[Chlamys farreri(Jones et Preston)](♀)和虾夷扇贝(Patinopecten yessoensis)(♂)杂交子代担轮幼虫为材料,分别用栉孔扇贝和虾夷扇贝基因组作探针,采用基因组荧光原位杂交(GISH)的方法,对杂交后代杂交子的确切身份进行初...以栉孔扇贝[Chlamys farreri(Jones et Preston)](♀)和虾夷扇贝(Patinopecten yessoensis)(♂)杂交子代担轮幼虫为材料,分别用栉孔扇贝和虾夷扇贝基因组作探针,采用基因组荧光原位杂交(GISH)的方法,对杂交后代杂交子的确切身份进行初步鉴定。结果表明,子代分别继承了双亲各一套染色体(n=19),为真正的杂交种。为了解杂交扇贝的免疫学特性,在自然海域栉孔扇贝大规模死亡的情况下,分别对杂交扇贝及其亲本3个扇贝群体血细胞的胞内活性氧含量(ROIs)、血清凝集素效价(HA)、溶菌酶活力(LSZ)、抑菌活力、酚氧化酶活性(PO)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)以及酸性磷酸酶(ACP)和碱性磷酸酶(ALP)等9种非特异性免疫学指标进行测定。结果表明,栉孔扇贝除ROIs、SOD、ACP等3个指标显著低于虾夷扇贝外(P<0.01),其他6种指标均高于虾夷扇贝,且除血清凝集素效价外均达显著水平(P<0.05)。在杂交子代中,上述免疫指标除SOD活性低于低值亲本外(P>0.6),其余8种免疫指标均介于双亲之间。杂交子代在9种免疫指标中有8种与母本无显著性差异,而子代与父本之间9种免疫指标中有7种达显著差异(P<0.05)。这些结果说明,杂交扇贝在非特异性免疫上存在明显的偏母性特征,这点与子代在外形特征上的偏母性相吻合。因此杂交扇贝相对于其母本在生产实践中表现出的一定程度的抗逆优势可能与非特异性免疫无明显关系。[中国水产科学,2006,13(4):597-602]展开更多
研究甘蔗属与滇蔗茅属间远缘杂交F1染色体遗传行为具有重要科学意义,可为发掘利用滇蔗茅野生优异基因资源提供细胞学依据。本研究采用常规压片技术对可育父本及不育杂交F1花粉母细胞减数分裂过程进行比较观察,结果显示可育父本云南95-1...研究甘蔗属与滇蔗茅属间远缘杂交F1染色体遗传行为具有重要科学意义,可为发掘利用滇蔗茅野生优异基因资源提供细胞学依据。本研究采用常规压片技术对可育父本及不育杂交F1花粉母细胞减数分裂过程进行比较观察,结果显示可育父本云南95-19减数分裂正常,而不育杂交F1分裂异常;进一步对F1花粉母细胞减数分裂过程进行基因组荧光原位杂交(GISH,genome in situ hybridization)分析,结果表明:滇蔗茅与甘蔗属热带种的亲缘关系较远,双亲染色体在F1细胞中不能进行同源配对,终变期,15条滇蔗茅染色体以单价体形式存在,花粉母细胞减数分裂细胞中存在滞后染色体、染色体丢失和不均衡分离现象。甘蔗-滇蔗茅属间远缘杂交F1花粉母细胞减数分裂异常因而杂交F1花粉完全败育。展开更多
基金the National Natural Science Foundation of China(No.30471222)
文摘Longiflorum and Asiatic lilies of the genus Lilium of the family Liliaceae are two important groups of modem lily cultivars. One of the main trends of lily breeding is to realize introgression between these groups. With cut style pollination and embryo rescue, distant hybrids between the two groups have been obtained. However, the FI hybrids are highly sterile or some of them could produce a small number of 2n gametes, and their BC1 progenies are usually triploids. Dutch lily breeders have selected many cultivars from these BC1 progenies based on their variation. It is presumably suggested that such variation could be caused by intergenomic recombination and abnormal meiosis during gamete formation in F1 hybrids of Longiflorum × Asiatic (LA) hybrids in Lilium. Therefore, the meiotic process of ten F1 LA hybrids was cytologically investigated using genomic in situ hybridization and traditional cytological methods in the present research. The results showed that: at metaphase I, the homoeologous chromosome pairing among different F1 hybrids ranged from 2.0 to 11.4 bivalents formed by homoeologous chromosomes per pollen mother cell (PMC), and very few multivalents, and even very few bivalents were formed by two chromosomes within one genome rather than homoeologous chromosomes in some PMCs; at anaphase I, all biva- lents were disjoined and most univalents were divided. Both the disjoined bivalents (half-bivalents) and the divided univalents (sister chromatids) moved to the opposite poles, and then formed two groups of chromosomes; because the two resulting half-bivalents retained their axes in the cell undisturbed, many crossover types, including single crossovers, three strand double crossovers, four strand double crossovers, four strand triple crossovers, and four strand multiple crossovers between the non-sister chromatids in the tetrads of bivalents, were clearly inferred by analyzing the breakpoints on the disjoined bivalents. The present investigation not only explained the reason for sterility of the Fl LA hybrids and the variation of their BCx progenies, but also provided a new method to analyze crossover types in other F1 interspecific hybrids as well.
基金The work was supported by National High Technology Development Program of China(Z16-02-01-02).
文摘To transfer desirable resistance traits from O. minuta to O. sativa, intergeneric hybrid plants between O. sativa (AA, 2n=2X=24) and O. minuta (BBCC, 2n=4X=48) were produced by embryo rescue after sexual cross. Morphological observation and chromosome counts indicated their hybrid status (ABC, 2n=3X=36). Genomic in situ hybridization (GISH) was further applied to confirm the parentage of the chromosomes of F 1 hybrids. Chromosomes of O. minuta and O. sativa were distinguishable in the hybrids in different fluorescence colors. GISH indicated that A and BC chromosomes were not randomly assembled in a cell. RAPD profiles unequivocally revealed their hybrids with double parent patterns. The results of blast tests showed that the hybrids had obtained disease resistance from O. minuta, and had a level of susceptibility between the parents.
文摘In the genus Oryza, interspecific hybrids are useful bridges for transferring the desired genes from wild species to cultivated rice (Oryza sativa L.). In the present study, hybrids between O. sativa (AA genome) and three Chinese wild rices, namely O. rufipogon (AA genome), O. officinalis (CC genome), and O. meyeriana (GG genome), were produced. Agricultural traits of the F1 hybrids surveyed were intermediate between their parents and appreciably resembled wild rice parents. Except for the O. sativa × O. rufipogon hybrid, the other F1 hybrids were completely sterile. Genomic in situ hybridization (GISH) was used for hybrid verification. Wild rice genomic DNAs were used as probes and cultivated rice DNA was used as a block. With the exception of O. rufipogon chromosomes, this method distinguished the other two wild rice and cultivated rice chromosomes at the stage of mitotic metaphase with different blocking ratios. The results suggest that a more distant phylogenetic relationship exists between O. meyeriana and O. sativa and that O. rufipogon and O. sativa share a high degree of sequence homology. The average mitotic chromosome length of O. officinalis and O. meyeriana was 1.25- and 1.51-fold that of O. sativa, respectively. 4',6'-Diamidino- 2-phenylindole staining showed that the chromosomes of O. officinalis and O. meyeriana harbored more heterochromatin, suggesting that the C and G genomes were amplified with repetitive sequences compared with the A genome. Although chromocenters formed by chromatin compaction were detected with wild rice-specific signals corresponding to the C and G genomes in discrete domains of the F1 hybrid interphase nuclei, the size and number of O. meyeriana chromocenters were bigger and greater than those of O. officinalis. The present results provide an important understanding of the genomic relationships and a tool for the transfer of useful genes from three native wild rice species in China to cultivars.
文摘Gossypium mustelinum [(AD)4] is one of five tetraploid species in Gossypium.Three pairs of nucleolar organizer regions(NOR) in(AD)4 were detected by FISH with 45S rDNA as a probe,they also were observed with genomic DNA(gDNA) from Gossypium D genome species as probes.Of
基金Supported by the National Natural Science Foundation of China (30671293).The authors thank Ms. Yanyan Ru and Mr. Wei Yue for their technical assistance and beneficial discussion on the manuscript.
文摘The St and E are two important basic genomes in the perennial tribe Triticeae (Poaceae). They exist in many perennial species and are very closely related to the A, B and D genomes of bread wheat (Triticum aestivum L.). Genomic Southern hybridization and genomic in situ hybridization (GISH) were used to analyze the genomic relationships between the two genomes (St and E) and the three basic genomes (A, B and D) of T. aestivum. The semi-quantitative analysis of the Southern hybridization suggested that both St and E genomes are most closely related to the D genome, then the A genome, and relatively distant to the B genome. GISH analysis using St and E genomic DNA as probes further confirmed the conclusion. St and E are the two basic genomes of Thinopyrum ponticum (StStE^eE^bE^x) and Th. intermedium (StE^eE^b), two perennial species successfully used in wheat improvement. Therefore, this paper provides a possible answer as to why most of the spontaneous wheat-Thinopyrum translocations and substitutions usually happen in the D genome, some in the A genome and rarely in the B genome. This would develop further use of alien species for wheat improvement, especially those containing St or E in their genome components.
基金the National NaturalScience Foundation of China (Grant No. 39870423) and the Doctorate Spot Fund of the Ministry of Education (Grant No. 207980112).
文摘Using the genomic DNAs of maize and rice as probes respectively, the homology of maize and rice genomes was assessed by genomic in situ hybridization. When rice genomic DNAs were hybridized to maize, all chromosomes displayed many multiple discrete regions, while each rice chromosome delineated a single consecutive chromosomal region after they were hybridized with maize genomic DNAs. The results indicate that the genomes of maize and rice share high homology, and confirm the proposal that maize and rice are diverged from a common ancestor.
基金the National Natural Science Foundation of China (30571184)Jiangsu Provincial Natural Science Foundation (BK2007166)+3 种基金the Tenth Five-year Plan of the National Key Program (2004BA525B05)the 111 Project(B08025) the Eleventh Five-year Plan of the National Sci-technologicalSupporting Program (2006BAD13B04-1-08)the Changjiang Scholars and Innovative Research Team in University and the Teaching and Research AwardProgram for Outstanding Young Teachers in Higher Education Institutions ofMinistry of Education (MOE), China.
文摘To identify alien chromosomes in recipient progenies and to analyze genome components in polyploidy, a genomic in situ hybridization (GISH) technique that is suitable for cotton was developed using increased stringency conditions. The increased stringency conditions were a combination of the four factors in the following optimized state: 100:1 ratio of blocking DNA to probe, 60% formamide wash solution, 43 ℃ temperature wash and a 13 min wash. Under these specific conditions using gDNA from Gossypium sturtianum (C1 C1 ) as a probe, strong hybridization signals were only observed on chromosomes from the C1 genome in somatic cells of the hybrid F1 (G. hirsutum x G. sturtianum) (AtDtC1). Therefore, GISH was able to discriminate parental chromosomes in the hybrid. Further, we developed a multi-color GISH to simultaneously discriminate the three genomes of the above hybrid. The results repeatedly displayed the three genomes, At, Dt, and C1, and each set of chromosomes with a unique color, making them easy to identify. The power of the multi-color GISH was proven by analysis of the hexaploid hybrid F1 (G. hirsutum x G. australe) (AtAtDtDtG2G2). We believe that the powerful multi-color GISH technique could be applied extensively to analyze the genome component in polyploidy and to identify alien chromosomes in the recipient progenies.
基金the National Natural Science Foundation of China (30671288and 30730065)
文摘Genomic in situ hybridization banding (GISH-banding), a technique slightly modified from conventional GISH, was used to probe the Chinese native rye (Secale cereale L.) DNA, and enabled us to visualize the individual rye chromosomes and create a universal reference karyotype of the S. cereale chromosome 1R to 7R. The GISH-banding approach used in the present study was able to discriminate S. cereale chromosomes or segments in the wheat (Triticum aestivum L.) background, including the Triticale, wheat-rye addition and translocation lines. Moreover, the GISH-banding pattern of S. cereale subsp. Afghanicum chromosomes was consistent with that of Chinese native rye cv. Jingzhou rye; whereas the GISH-banding pattern of Secale vavilovii was different from that of S. cereale, indicating that GISH-banding can be used to study evolutionary polymorphism in species or subspecies of Secale. In addition, the production and application of GISH-banding to the study of adenine-thymine-riched heterochromaUn is discussed.
文摘以栉孔扇贝[Chlamys farreri(Jones et Preston)](♀)和虾夷扇贝(Patinopecten yessoensis)(♂)杂交子代担轮幼虫为材料,分别用栉孔扇贝和虾夷扇贝基因组作探针,采用基因组荧光原位杂交(GISH)的方法,对杂交后代杂交子的确切身份进行初步鉴定。结果表明,子代分别继承了双亲各一套染色体(n=19),为真正的杂交种。为了解杂交扇贝的免疫学特性,在自然海域栉孔扇贝大规模死亡的情况下,分别对杂交扇贝及其亲本3个扇贝群体血细胞的胞内活性氧含量(ROIs)、血清凝集素效价(HA)、溶菌酶活力(LSZ)、抑菌活力、酚氧化酶活性(PO)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)以及酸性磷酸酶(ACP)和碱性磷酸酶(ALP)等9种非特异性免疫学指标进行测定。结果表明,栉孔扇贝除ROIs、SOD、ACP等3个指标显著低于虾夷扇贝外(P<0.01),其他6种指标均高于虾夷扇贝,且除血清凝集素效价外均达显著水平(P<0.05)。在杂交子代中,上述免疫指标除SOD活性低于低值亲本外(P>0.6),其余8种免疫指标均介于双亲之间。杂交子代在9种免疫指标中有8种与母本无显著性差异,而子代与父本之间9种免疫指标中有7种达显著差异(P<0.05)。这些结果说明,杂交扇贝在非特异性免疫上存在明显的偏母性特征,这点与子代在外形特征上的偏母性相吻合。因此杂交扇贝相对于其母本在生产实践中表现出的一定程度的抗逆优势可能与非特异性免疫无明显关系。[中国水产科学,2006,13(4):597-602]
文摘研究甘蔗属与滇蔗茅属间远缘杂交F1染色体遗传行为具有重要科学意义,可为发掘利用滇蔗茅野生优异基因资源提供细胞学依据。本研究采用常规压片技术对可育父本及不育杂交F1花粉母细胞减数分裂过程进行比较观察,结果显示可育父本云南95-19减数分裂正常,而不育杂交F1分裂异常;进一步对F1花粉母细胞减数分裂过程进行基因组荧光原位杂交(GISH,genome in situ hybridization)分析,结果表明:滇蔗茅与甘蔗属热带种的亲缘关系较远,双亲染色体在F1细胞中不能进行同源配对,终变期,15条滇蔗茅染色体以单价体形式存在,花粉母细胞减数分裂细胞中存在滞后染色体、染色体丢失和不均衡分离现象。甘蔗-滇蔗茅属间远缘杂交F1花粉母细胞减数分裂异常因而杂交F1花粉完全败育。