毛竹和厚壁毛竹的核型分析及45S和5S rDNA的FISH分析
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摘要
【目的】厚壁毛竹(Phyllostachys edulis‘Pachyloen’)是毛竹(Phyllostachys edulis)的栽培品种,是具有较高经济价值的优特种质。对毛竹与厚壁毛竹进行细胞遗传学研究,了解其核型特征。【方法】采用双色荧光原位杂交技术,对毛竹、厚壁毛竹进行了核型分析,利用45S和5S rDNA在其染色体上进行了物理定位。【结果】毛竹、厚壁毛竹的染色体数目都为48条,毛竹的染色体相对组成为2n=48=12L+10M_2+14M_1+12S,核型公式为2n=48=32m+12sm+4st(2SAT);厚壁毛竹的染色体相对组成为2n=48=12L+8M_2+16M_1+12S,核型公式为2n=48=34m+10sm+4st(2SAT)。毛竹与厚壁毛竹的染色体不对称系数分别为61.41%和59.55%,均属于"2B"型。荧光原位杂交(FISH)结果表明:毛竹、厚壁毛竹都有2个45S rDNA位点和4个5S rDNA位点。【结论】厚壁毛竹的染色体数量为48条,补充了毛竹的染色体为48条的循证依据;获得了毛竹、厚壁毛竹45S和5S rDNA荧光原位杂交核型,两者的45S和5S rDNA位点没有明显差异,具有相似的核型,显示其较近的亲缘关系。
【Objective】Phyllostachys edulis ‘Pachyloen' is a cultivar of Phyllostachys edulis, which is an economically-important germplasm. Cytogenetic research on this germplasm provide a cytomolecular basis for genetic research on bamboo species.【Method】Karyotype analysis and physical localization of 45 S and 5 S rDNA on the chromosomes of Ph. edulis and Ph. edulis ‘Pachyloen' were performed by double color fluorescence in situ hybridization(FISH).【Result】The results of karyotype analysis indicated that the chromosome number in the two materials was 2 n=48, the karyotype formula of Ph. edulis and Ph. edulis ‘Pachyloen' was 2 n=48=32 m+12 sm+4 st(2 SAT) and 2 n=48=34 m+10 sm+4 st(2 SAT), respectively. The chromosome composition of relative length was 2 n=48=12 L+10 M2+14 M1+12 S and 2 n=48=12 L+8 M2+16 M1+12 S, respectively. The asymmetrical karyotype coefficients of Ph. edulis and Ph. edulis ‘Pachyloen' were 61.41% and 59.55%, and their karyotypes were both of 2 B type. The results showed that there were 2 loci of 45 S rDNA and 4 loci of 5 S rDNA in both Ph. edulis and Ph. edulis ‘Pachyloen'. 【Conclusion】The chromosome number in the two materials is 2 n=48, the 45 S and 5 S rDNA loci of the two materials have no obvious differences, and the two varieties have similar karyotypes, which shows that they have close genetic relatedness.
【Objective】Phyllostachys edulis ‘Pachyloen' is a cultivar of Phyllostachys edulis, which is an economically-important germplasm. Cytogenetic research on this germplasm provide a cytomolecular basis for genetic research on bamboo species.【Method】Karyotype analysis and physical localization of 45 S and 5 S rDNA on the chromosomes of Ph. edulis and Ph. edulis ‘Pachyloen' were performed by double color fluorescence in situ hybridization(FISH).【Result】The results of karyotype analysis indicated that the chromosome number in the two materials was 2 n=48, the karyotype formula of Ph. edulis and Ph. edulis ‘Pachyloen' was 2 n=48=32 m+12 sm+4 st(2 SAT) and 2 n=48=34 m+10 sm+4 st(2 SAT), respectively. The chromosome composition of relative length was 2 n=48=12 L+10 M2+14 M1+12 S and 2 n=48=12 L+8 M2+16 M1+12 S, respectively. The asymmetrical karyotype coefficients of Ph. edulis and Ph. edulis ‘Pachyloen' were 61.41% and 59.55%, and their karyotypes were both of 2 B type. The results showed that there were 2 loci of 45 S rDNA and 4 loci of 5 S rDNA in both Ph. edulis and Ph. edulis ‘Pachyloen'. 【Conclusion】The chromosome number in the two materials is 2 n=48, the 45 S and 5 S rDNA loci of the two materials have no obvious differences, and the two varieties have similar karyotypes, which shows that they have close genetic relatedness.
引文
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[10] 张莹,田埂,路慧萍,等.厚壁毛竹六个节气笋芽发育的转录组分析[J].江西农业大学学报,2015,37(3):466-474.DOI:10.13836/j.jjau.2015073.ZHANG Y,TIAN G,LU H P,et al.Transcriptome characterization of Phyllostachys edulis ‘Pachyloen’ shoots in different solar terms[J].Acta Agriculturae Universitatis Jiangxiensis,2015,37(3):466-474.
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[12] 徐川梅,卢江杰,汤定钦.45S rDNA在7种竹子植物染色体上的定位[J].林业科学,2009,45(12):42-45.DOI:10.3321/j.issn:1001-7488.2009.12.007.XU C M,LU J J,TANG D Q.Physical mapping of the 45S rDNA on metaphase chromosomes in seven bamboo species by in situ hybridization[J].Scientia Silvae Sinicae,2009,45(12):42-45.
[13] 轩淑欣,申书兴,赵建军,等.25S rDNA和5S rDNA在大白菜中期染色体上的FISH定位[J].中国农业科学,2007,40(4):782-787.DOI:10.3321/j.issn:0578-1752.2007.04.018.XUAN S X,SHEN S X,ZHAO J J,et al.Location of 25S rDNA and 5S rDNA in Chinese cabbage-pe-tsai metaphase chromosome[J].Scientia Agricultura Sinica,2007,40(4):782-787.
[14] JIANG J M,GILL B S.Current status and the future of fluorescence in situ hybridization (FISH) in plant genome research[J].Genome,2006,49(9):1057-1068.DOI:10.1139/g06-076.
[15] HAN Y H,ZHANG T,THAMMAPICHAI P,et al.Chromosome-specific painting in cucumis species using bulked oligonucleotides[J].Genetics,2015,200(3):771-779.DOI:10.1534/genetics.115.177642.
[16] SOLTIS D E,SOLTIS P S.Polyploidy:recurrent formation and genome evolution[J].Trends in Ecology & Evolution,1999,14(9):348-352.DOI:10.1016/s0169-5347(99)01638-9.
[17] VOLKOV R A,KOMAROVA N Y,HEMLEBEN V.Ribosomal DNA in plant hybrids:inheritance,rearrangement,expression[J].Systematics and Biodiversity,2007,5(3):261-276.DOI:10.1017/s1477200007002447.
[18] JIANG J M,GILL B S.Sequential chromosome banding and in situ hybridization analysis[J].Genome,1993,36(4):792-795.DOI:10.1139/g93-104.
[19] 陈佩度,周波,齐莉莉,等.用分子原位杂交(GISH)鉴定小麦-簇毛麦双倍体、附加系、代换系和易位系[J].遗传学报,1995,22(5):380-386.CHEN P D,ZHOU B,QI L L,et al.Identification of wheat-haynaldia villosa amphiploid,addition,substitution and translocation lines by in situ hybridization using biotin-labelled genomic DNA as a probe[J].Journal of Genetics and Genomics,1995,22(5):380-386.
[20] 李懋学,陈瑞阳.关于植物核型分析的标准化问题[J].植物科学学报,1985,3(4):297-302.LI M X,CHEN R Y.A suggestion on the standarization of karyotype analysis in plant[J].Journal of Plant Science,1985,3(4):297-302.
[21] LEVAN A,FREDGA K,SANDBERG A A.Nomenclature for centromeric position on chromosomes[J].Hereditas,2009,52(2):201-220.DOI:10.1111/j.1601-5223.1964.tb01953.x.
[22] KUO S R,WANG T T,HUANG T C.Karyotype analysis of some Formosan gymnosperms[J].Taiwania,1972,17(1):66-80.
[23] STEBBINS G L.Chromosomal evolution in higher plants[M].London:Edward Arnold (Publishers) Ltd,1971.
[24] ARANO H.Cytological studies in subfamily carduoideae (compositae) of Japan VIII[J].Shokubutsugaku Zasshi,1962,75(892):401-410.DOI:10.15281/jplantres1887.75.401.
[25] 刘大钧.细胞遗传学[M].北京:中国农业出版社,1999:9-10.LIU D J.Cylogenetics[M].Beijing:China Agyisurtural Publishing House,1999:9-10.
[2] ZHANG Y J,MA P F,LI D Z.High-throughput sequencing of six bamboo chloroplast genomes:phylogenetic implications for temperate woody bamboos (Poaceae:Bambusoideae)[J].PLoS One,2011,6(5):e20596.
[3] 李秀兰,林汝顺,冯学琳,等.中国部分丛生竹类染色体数目报道[J].植物分类学报,2001(5):433-442,T001-T002.LI X L,LIN R S,FUNG H L,et al.Chromosome numbers of some caespitose bamboosnative in or introduced to China[J].Acta Phytotaxonomica Sinica,2001(5):433-442.
[4] 林树燕,石文文,缪彬彬,等.竹类植物生殖生物学研究进展[J].世界竹藤通讯,2010,8(2):1-6.DOI:10.3909/j.issn.1672-0431.2010.02.001.LIN S Y,SHI W W,MIU B B,et al.Research advances in reproduction biology of bamboos[J].World Bamboo and Rattan,2010,8(2):1-6.
[5] 杨光耀,黎祖尧,杜天真,等.毛竹新栽培变种——厚皮毛竹[J].江西农业大学学报,1997,19(04):97-98.YANG G Y,LI Z Y,DU T Z,et al.A new cultivated variety of moso bamboo[J].Journal of Jiangxi Agricultural University,1997,19(4):97-98.
[6] 魏强,高志鹏,陈铭,等.厚壁毛竹与毛竹地下茎发育规律的比较[J].南京林业大学学报(自然科学版),2018,42(2):197-201.DOI:10.3969/j.issn.1000-2006.201611021.WEI Q,GAO Z P,CHEN M,et al.Morphological and cellular characterization of rhizome development of Phyllostachys edulis ‘Pachyloen’ and Phyllostachys edulis[J].Journal of Nanjing Forestry University(Natural Sciences Edition),2018,42(2):197-201.
[7] 丁雨龙.刚竹属(Phyllostachys)系统分类的研究[D].南京:南京林业大学,1998.DING Y L.Studies on taxonomy of Phyllostachys [D].Nanjing:Nanjing Forestry University,1998.
[8] 郭起荣.厚壁毛竹的种质性状及其繁育研究[D].长沙:中南林学院,2003.GUO Q R.Studies on germplasm characters and breeding of Phyllostachys edulis cv.pachyloen[D].Changsha:Central South Forestry College,2003.
[9] 李建,杨清培,方楷,等.厚壁毛竹与毛竹的抗寒生理比较研究[J].江西农业大学学报,2011,33(3):537-541.DOI:10.3969/j.issn.1000-2286.2011.03.023.LI J,YANG Q P,FANG K,et al.Studies on physiological characters of cold resistance of Phyllostachys edulis ‘ Pachyloen’ and Phyllostachys edulis[J].Acta Agriculturae Universitatis Jiangxiensis(Natural Sciences Edition),2011,33(3):537-541.
[10] 张莹,田埂,路慧萍,等.厚壁毛竹六个节气笋芽发育的转录组分析[J].江西农业大学学报,2015,37(3):466-474.DOI:10.13836/j.jjau.2015073.ZHANG Y,TIAN G,LU H P,et al.Transcriptome characterization of Phyllostachys edulis ‘Pachyloen’ shoots in different solar terms[J].Acta Agriculturae Universitatis Jiangxiensis,2015,37(3):466-474.
[11] 陈瑞阳.中国主要经济植物基因组染色体图谱——第Ⅱ册[M].北京:科学出版社,2003:484-485.CHEN R Y.Chromosome atlas of major economic plants genome in China:second volume [M].Beijing:Science Press,2003:484-485.
[12] 徐川梅,卢江杰,汤定钦.45S rDNA在7种竹子植物染色体上的定位[J].林业科学,2009,45(12):42-45.DOI:10.3321/j.issn:1001-7488.2009.12.007.XU C M,LU J J,TANG D Q.Physical mapping of the 45S rDNA on metaphase chromosomes in seven bamboo species by in situ hybridization[J].Scientia Silvae Sinicae,2009,45(12):42-45.
[13] 轩淑欣,申书兴,赵建军,等.25S rDNA和5S rDNA在大白菜中期染色体上的FISH定位[J].中国农业科学,2007,40(4):782-787.DOI:10.3321/j.issn:0578-1752.2007.04.018.XUAN S X,SHEN S X,ZHAO J J,et al.Location of 25S rDNA and 5S rDNA in Chinese cabbage-pe-tsai metaphase chromosome[J].Scientia Agricultura Sinica,2007,40(4):782-787.
[14] JIANG J M,GILL B S.Current status and the future of fluorescence in situ hybridization (FISH) in plant genome research[J].Genome,2006,49(9):1057-1068.DOI:10.1139/g06-076.
[15] HAN Y H,ZHANG T,THAMMAPICHAI P,et al.Chromosome-specific painting in cucumis species using bulked oligonucleotides[J].Genetics,2015,200(3):771-779.DOI:10.1534/genetics.115.177642.
[16] SOLTIS D E,SOLTIS P S.Polyploidy:recurrent formation and genome evolution[J].Trends in Ecology & Evolution,1999,14(9):348-352.DOI:10.1016/s0169-5347(99)01638-9.
[17] VOLKOV R A,KOMAROVA N Y,HEMLEBEN V.Ribosomal DNA in plant hybrids:inheritance,rearrangement,expression[J].Systematics and Biodiversity,2007,5(3):261-276.DOI:10.1017/s1477200007002447.
[18] JIANG J M,GILL B S.Sequential chromosome banding and in situ hybridization analysis[J].Genome,1993,36(4):792-795.DOI:10.1139/g93-104.
[19] 陈佩度,周波,齐莉莉,等.用分子原位杂交(GISH)鉴定小麦-簇毛麦双倍体、附加系、代换系和易位系[J].遗传学报,1995,22(5):380-386.CHEN P D,ZHOU B,QI L L,et al.Identification of wheat-haynaldia villosa amphiploid,addition,substitution and translocation lines by in situ hybridization using biotin-labelled genomic DNA as a probe[J].Journal of Genetics and Genomics,1995,22(5):380-386.
[20] 李懋学,陈瑞阳.关于植物核型分析的标准化问题[J].植物科学学报,1985,3(4):297-302.LI M X,CHEN R Y.A suggestion on the standarization of karyotype analysis in plant[J].Journal of Plant Science,1985,3(4):297-302.
[21] LEVAN A,FREDGA K,SANDBERG A A.Nomenclature for centromeric position on chromosomes[J].Hereditas,2009,52(2):201-220.DOI:10.1111/j.1601-5223.1964.tb01953.x.
[22] KUO S R,WANG T T,HUANG T C.Karyotype analysis of some Formosan gymnosperms[J].Taiwania,1972,17(1):66-80.
[23] STEBBINS G L.Chromosomal evolution in higher plants[M].London:Edward Arnold (Publishers) Ltd,1971.
[24] ARANO H.Cytological studies in subfamily carduoideae (compositae) of Japan VIII[J].Shokubutsugaku Zasshi,1962,75(892):401-410.DOI:10.15281/jplantres1887.75.401.
[25] 刘大钧.细胞遗传学[M].北京:中国农业出版社,1999:9-10.LIU D J.Cylogenetics[M].Beijing:China Agyisurtural Publishing House,1999:9-10.