rDNA在斑茅染色体的物理定位和染色体基数研究
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摘要
为了了解5S rDNA和45S rDNA在斑茅染色体上的分布情况及其染色体基数,利用荧光原位杂交技术在斑茅海南92-77(2n=60)和海南92-105(2n=60)的中期染色体和间期细胞核上进行5S rDNA和45S rDNA定位研究。结果表明在海南92-77和海南92-105的中期染色体和间期细胞核上45S rDNA位点均有6个信号位点,且均位于染色体短臂的端部上;这6个45S rDNA位点大小具有明显的差异,说明斑茅的45S rDNA的拷贝数存在差异。5S rDNA位点在海南92-77和海南92-105均有6个信号位点,且均位于染色体长臂的着丝粒附近;5S rDNA位点大小相差无几,说明斑茅的5S rDNA的拷贝数差异不大。因此,可以推断染色体数目为60的斑茅染色体基数为x=10。
In order to know the physical mapping of 5 S rDNA and 45 S rDNA on the chromosomes of E. arundinaceus and determination of basic chromosome number, the E. arundinaceus chromosome localization of 5 S rDNA and 45 S rDNA was studied by fluorescence in situ hybridization on metaphase chromosomes in Hainan 92-77(2 n=60) and Hainan 92-105(2 n=60). The results showed that the number of 45 S rDNA site signal loci were six at somatic chromosomes of metaphase and cell nucleus of interphase in both Hainan 92-77 and Hainan 92-105, and the signal loci of 45 S rDNA was located in the short arm end of six chromosomes. Among them, loci were different, suggesting that the copy number of 45 S rDNA was different. The number of 5 S rDNA site signal loci were six in both Hainan 92-77 and Hainan 92-105, and the signal loci of 45 S rDNA was located near the long arm of chromosome centromere, all the 6 loci were of similar size, suggesting that the copy number of 5 S rDNA was similar. Therefore, the basic chromosome number of E. arundinaceus with 2 n=60 was determined as x=10.
In order to know the physical mapping of 5 S rDNA and 45 S rDNA on the chromosomes of E. arundinaceus and determination of basic chromosome number, the E. arundinaceus chromosome localization of 5 S rDNA and 45 S rDNA was studied by fluorescence in situ hybridization on metaphase chromosomes in Hainan 92-77(2 n=60) and Hainan 92-105(2 n=60). The results showed that the number of 45 S rDNA site signal loci were six at somatic chromosomes of metaphase and cell nucleus of interphase in both Hainan 92-77 and Hainan 92-105, and the signal loci of 45 S rDNA was located in the short arm end of six chromosomes. Among them, loci were different, suggesting that the copy number of 45 S rDNA was different. The number of 5 S rDNA site signal loci were six in both Hainan 92-77 and Hainan 92-105, and the signal loci of 45 S rDNA was located near the long arm of chromosome centromere, all the 6 loci were of similar size, suggesting that the copy number of 5 S rDNA was similar. Therefore, the basic chromosome number of E. arundinaceus with 2 n=60 was determined as x=10.
引文
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[20]LAN H,CHEN C L,MIAO Y,et al.Fragile sites of‘Valencia’sweet orange(Citrus sinensis)chromosomes are related with active 45S rDNA[J].PLo S One,2016,11(3):e0151512.
[21]FERRO D A,NéO D M,MOREIRA-FILHO O,et al.Nucleolar organizing regions,18S and 5S rDNA in Astyanax scabripinnis(Pisces,Characidae):populations distribution and functional diversity[J].Genetica,2000,110(1):55-62.
[22]HUSSEIN M,EL-TWAB A.Physical mapping of the 45S rDNA on the chromosomes of Triticum turgidum and T.aestivum using fluorescence in situ hybridization for chromosome ancestors[J].Arab J Biotech,2007,10(10):69-80.
[23]ZHANG H,BIAN Y,GOU X,et al.Persistent whole-chromosome aneuploidy is generally associated with nascent allohexaploid wheat[J].Proc Natl Acad Sci U S A,2013,110(9):3447-3452.
[24]徐川梅,别同德,王春梅,等.45S rDNA在小麦及其近缘物种染色体上的分布[J].遗传,2007,29(9):1126-1130.
[25]IRVINE J.Saccharum species as horticultural classes[J].Theor Appl Genet,1999,98(2):186-194.
[26]SINGH R K,SINGH R B,SINGH S P,et al.Identification of sugarcane microsatellites associated to sugar content in sugarcane and transferability to other cereal genomes[J].Euphytica,2011,182(3):335-354.
[2]DE ALMEIDATOLEDO L F,STOCKER A J,FORESTI F,et al.Fluorescence in situ hybridization with rDNA probes on chromosomes of two nucleolus organizer region phenotypes of a species of Eigenmannia(Pisces,Gymnotoidei,Sternopygidae)[J].Chromosome Res,1996,4(4):301-305.
[3]CHEN G F,ZHANG W C Y,ZHANG B Y,et al.Development of r RNA and rDNA-targeted probes for fluorescence in situ hybridization to detect Heterosigma akashiwo(Raphidophyceae)[J].J Exp Mar Biol Ecol,2008,355(1):66-75.
[4]FUKUI K,OHMIDO N,KHUSH G S.Variability in rDNA loci in the genus Oryza detected through fluorescence in situ hybridization[J].Theor Appl Genet,1994,87(8):893-899.
[5]D’HONT A,ISON D,ALIX K,et al.Determination of basic chromosome numbers in the genus Saccharum by physical mapping of ribosomal RNA genes[J].Genome,1998,41(2):221-225.
[6]D’HONT A,RAO P,FELDMANN P,et al.Identification and characterization of sugarcane intergeneric hybrids,Saccharum officinarum×Erianthus arundinaceus,with molecular markers and DNA in situ hybridization[J].Theor Appl Genet,1995,91(2):320-326.
[7]JENKIN M J,READER S M,PURDIE K A,et al.Detection of rDNA sites in sugarcane by FISH[J].Chromosome Res,1995,3(7):444-445.
[8]BESSE P,MCINTYRE C L.Chromosome in situhybridization of ribosomal DNA in Erianthus sect.R[J].Genome,1999,42(2):270-273.
[9]LU Y,D’HONT A,WALKER D,et al.Relationships among ancestral species of sugarcane revealed with RFLP using single copy maize nuclear probes[J].Euphytica,1994,78(1-2):7-18.
[10]IRVINE J.Saccharum species as horticultural classes[J].Theor Appl Genet,1999,98(2):186-194.
[11]AMALRAJ V A,BALASUNDARAM N.On the taxonomy of the members of‘Saccharum complex’[J].Genet Resour Crop Ev,2006,53(1):35-41.
[12]ROTT P,MOHAMED I,KLETT P,et al.Resistance to leaf scald disease is associated with limited colonization of sugarcane and wild relatives by Xanthomonas albilineans[J].Phytopathology,1997,87(12):1202-1213.
[13]PIPERIDIS G,CHRISTOPHER M J,CARROLL B J,et al.Molecular contribution to selection of intergeneric hybrids between sugarcane and the wild species Erianthus arundinaceus[J].Genome,2000,43(6):1033-1037.
[14]RAM B,SREENIVASAN T,SAHI B,et al.Introgression of low temperature tolerance and red rot resistance from Erianthus in sugarcane[J].Euphytica,2001,122(1):145-153.
[15]CAI Q,AITKEN K S,FAN Y H,et al.A preliminary assessment of the genetic relationship between Erianthusrockii and the“Saccharum complex”using microsatellite(SSR)and AFLP markers[J].Plant Sci,2005,169(5):976-984.
[16]TAI P,MILLER J.A Core Collection for Saccharum spontaneum L.from the World Collection of Sugarcane[J].Crop Sci,2001,41(3):879-885.
[17]TAI P,MILLER J.Germplasm diversity among four sugarcane species for sugar composition[J].Crop Sci,2002,42(3):958-964.
[18]JACKSON P,HENRY R J.Erianthus[M].Berlin:Springer Berlin Heidelberg,2011:97-107.
[19]杨清辉,李富生,肖凤回,等.斑茅染色体和植物学性状观察研究[J].云南农业大学学报,1997,12(4):253-256.
[20]LAN H,CHEN C L,MIAO Y,et al.Fragile sites of‘Valencia’sweet orange(Citrus sinensis)chromosomes are related with active 45S rDNA[J].PLo S One,2016,11(3):e0151512.
[21]FERRO D A,NéO D M,MOREIRA-FILHO O,et al.Nucleolar organizing regions,18S and 5S rDNA in Astyanax scabripinnis(Pisces,Characidae):populations distribution and functional diversity[J].Genetica,2000,110(1):55-62.
[22]HUSSEIN M,EL-TWAB A.Physical mapping of the 45S rDNA on the chromosomes of Triticum turgidum and T.aestivum using fluorescence in situ hybridization for chromosome ancestors[J].Arab J Biotech,2007,10(10):69-80.
[23]ZHANG H,BIAN Y,GOU X,et al.Persistent whole-chromosome aneuploidy is generally associated with nascent allohexaploid wheat[J].Proc Natl Acad Sci U S A,2013,110(9):3447-3452.
[24]徐川梅,别同德,王春梅,等.45S rDNA在小麦及其近缘物种染色体上的分布[J].遗传,2007,29(9):1126-1130.
[25]IRVINE J.Saccharum species as horticultural classes[J].Theor Appl Genet,1999,98(2):186-194.
[26]SINGH R K,SINGH R B,SINGH S P,et al.Identification of sugarcane microsatellites associated to sugar content in sugarcane and transferability to other cereal genomes[J].Euphytica,2011,182(3):335-354.