Estimation of nuclear DNA content of 17 Chinese wild rose species by flow cytometry
-
摘要: 以17种中国野生蔷薇为试材，采用改良的LB 01裂解液，以4种不同的标准植物——大豆（Glycine max Merr.‘Polanka’）、绿豆（Vigna radiata（L.） Wilczek）、番茄（Lycopersicon esculentum Miller）和玉米（Zea mays L.）为外标，以二倍体材料丽江蔷薇（Rosa×lichiangensis Yü et Ku）为内部参照，利用流式细胞术对其核DNA含量及染色体倍性进行检测，并采用常规染色体压片法验证倍性准确性。本研究首次检测了3个二倍体种——商城蔷薇（Rosa shangchengensis T.C.Ku）、广东蔷薇（Rosa kwangtungensis Yü et Tsai）和无刺刺梨（Rosa roxburghii f.inermis S.D.Shi），1个三倍体种——伞房蔷薇（Rosa corymbulosa Rolfe）和1个四倍体种——弯刺蔷薇（Rosa beggeriana Schrenk）的核DNA含量及基因组大小。结果表明，流式细胞术检测结果与常规染色体压片法结果一致，可对中国野生蔷薇的倍性研究进行补充。本研究结果可丰富中国蔷薇属植物的细胞遗传学背景资料并为繁育新品种提供理论依据。Abstract: Using four different standard plants (Glycine max Merr.‘Polanka’,Vigna radiata(L.) Wilczek,Lycopersicon esculentum Miller,and Zea mays L.) as outer calibrations and diploid taxon Rosa×lichiangensis Yü et Ku as the inner calibration,we estimated the nuclear genome sizes of 17 wild roses in China using flow cytometry based on the improved LB 01 buffer.Simultaneously,traditional chromosome tableting was performed to verify ploidy levels.The nuclear DNA contents and genome sizes of three diploid species (Rosa shangchengensis T.C.Ku,Rosa kwangtungensis Yü et Tsai and Rosa roxburghii f.inermis S.D.Shi),one triploid species (Rosa corymbulosa Rolfe),and one tetraploid species (Rosa beggeriana Schrenk) were reported in this study for the first time.The determined ploidy levels based on flow cytometry were consistent with those obtained by the traditional chromosome tableting technique.These findings enrich the cytogenetic resources of Rosa L.in China and offer a basis for the breeding of new strains.
-
Keywords:
- Rosa /
- Nuclear DNA content /
- Ploidy level /
- Flow cytometry
-
-
[1] Ku TC, Robertson KR. Rosa(Rosaceae)[M]//Wu ZY, Raven PH, eds. Flora of China. Vol 9. Beijing:Science Press; Saint Louis:Missouri Botanical Garden Press, 2003:339-381.
[2] Jian HY, Zhang H, Tang KX, Li SF, Wang Q, et al. Decaploidy in Rosa praelucens Byhouwer (Rosaceae) endemic to Zhongdian Plateau, Yunnan, China[J]. Caryologia, 2010, 63(2):162-167.
[3] 田新民, 周香艳, 弓娜. 流式细胞术在植物学研究中的应用——检测植物核DNA含量和倍性水平[J]. 中国农学通报, 2011, 27(9):21-27. Tian XM, Zhou XY, Gong N. Application of flow cytometry in plant research-analysis of nuclear DNA content and ploidy level in plant cells[J]. Chinese Agricultural Science Bulletin, 2011, 27(9):21-27.
[4] Greilhuber J, Doležel J, Lysak MA, Bennett MD. The origin, evolution and proposed stabilization of the terms ‘Genome Size’ and ‘C-Value’ to describe nuclear DNA contents[J]. Ann Bot, 2005, 95(1):255-260.
[5] Dickson EE, Arumuganathan K, Kresovich S, Doyle JJ. Nuclear DNA content variation within the Rosaceae[J]. Am J Bot, 1992:1081-1086.
[6] Yokoya K, Roberts AV, Mottley J, Lewis R, Brandham PE. Nuclear DNA amounts in roses[J].[QX(Y12#] Ann Bot, 2000, 85(4):557-561.
[7] Roberts AV, Gladis T, Brumme H. DNA amounts of roses (RosaL.) and their use in attributing ploidy levels[J]. Plant Cell Rep, 2009, 28(1):61-71.
[8] Jian HY, Zhang T, Wang Q, Yan HJ, Qiu XQ, et al. Nuclear DNA content and 1Cx-value variations in genus RosaL.[J]. Caryologia, 2014, 67(4):273-280.
[9] 武荣花, 葛蓓蓓, 王茂良, 周燕, 冯慧. 应用流式细胞术测18个中国古老月季基因组大小[J]. 北京林业大学学报, 2016, 38(6):98-104. Wu RH, Ge BB, Wang ML, Zhou Y, Feng H. Estimation of genome size of eighteen Chinese old garden roses by flow cytometry[J]. Journal of Beijing Forestry University, 2016, 38(6):98-104.
[10] Doležel J, Doleželová M, Novák FJ. Flow cytometric estimation of nuclear DNA amount in diploid bananas (Musa acuminata, and M. balbisiana)[J]. Biol Plantarum, 1994, 36(3):351-357.
[11] Bennett MD, Smith JB. Nuclear DNA amounts in angiosperms[J]. Philos T R Soc B, 1991, 334(1271):309-345.
[12] Doležel J, Sgorbati S, Lucretti S. Comparison of three DNA fluorochromes for flow cytometric estimation of nuc-lear DNA content in plants[J]. Physiol Plantarum, 1992, 85(4):625-631.
[13] Lysak MA, Doležel J. Estimation of nuclear DNA content in Sesleria(Poaceae)[J]. Caryologia, 1998, 51(2):123-132.
[14] Doležel J, Greilhuber J, Suda J. Estimation of nuclear DNA content in plants using flow cytometry[J]. Nat Protoc, 2007, 2(9):2233-2244.
[15] 马燕, 陈俊愉. 中国蔷薇属6个种的染色体研究[J]. 广西植物, 1992, 12(4):333-336. Ma Y, Chen JY. Chromosome studies of 6 species of Rosain China[J]. Guihaia, 1992, 12(4):333-336.
[16] Popek R, Facsar G, Malecka J. Cyto-taxonomische untersuchungen an der gattung Rosa(Rosaceae)-die arten aus ungarn und anderen gebieten[J]. Fragm Flor Geobot, 1991, 36(1):81-87.
[17] Sedelnikova TS, Pimenov AV. Cytogenetic features of woody plants adaptation in extreme environmental conditions[J]. Bull State Nikitsky Gardens, 2002, 86:61-62.
[18] Zhou XY, Ma JF, Wang WY, Gong N, Zhang YY, Liu JQ. Genome size of the diploid hybrid species Hippophae goniocarpaand its parental species, H. rhamnoidesssp. sinensisand H. neurocarpassp. neurocarpa(Elaeagnaceae)[J]. Acta Biol Cracov Bot, 2010, 52(2):12-16.
[19] 吴青青, 赖彪, 朱文超, 钟沛霖. 流式细胞术检测草莓倍性的方法优化[J]. 现代农业科技, 2015(20):48-50. Wu QQ, Lai B, Zhu WC, Zhong PL. Method optimization for detection of strawberry ploidy by flow cytometry[J]. Modern Agricultural Science and Technology, 2015(20):48-50.
[20] 富新年, 高洪文, 王赞. 流式细胞术鉴定黄花苜蓿染色体倍性水平[J]. 中国草地学报, 2013,35(1):18-22. Fu XN, Gao HW, Wang Z. Ploidy levels of Medicago falcategermplasm determined by flow cytometry[J]. Chinese Journal of Grassland, 2013, 35(1):18-22.
[21] 吴青青, 胡小京, 黄伟, 杨雪莲, 夏景风, 鲁周. 流式细胞仪测定百合花粉倍性的研究[J]. 安徽农业科学, 2015, 43(28):4-6. Wu QQ, Hu XJ, Huang W, Yang XL, Xia JF, Lu Z. Study on determination of Liliumpollen ploidy by flow cytometry[J]. Journal of Anhui Agricultural Sciences, 2015, 43(28):4-6.
[22] Zhang NN, Yang J, Sun WB. Genome size estimation of Viburnum(Adoxaceae) species by using flow cytometry[J]. Plant Diversity and Resources, 2014, 36(6):730-736.
[23] 张志珂, 王永清, 林顺权, 杜奎. 借助细胞流式仪进行枇杷基因组学测序材料的倍性鉴定[J]. 果树学报, 2012, 29(3):498-504. Zhang ZK, Wang YQ, Lin SQ, Du K. Ploidy identification of loquats for genome sequencing project by flow cytometry[J]. Journal of Fruit Science, 2012, 29(3):498-504.
[24] 魏高明, 方炎明. 栎属植物流式细胞术实验体系的建立与优化[J]. 南京林业大学学报:自然科学版, 2015, 39(1):167-172. Wei GM, Fang YM. Establishment and optimization of an experiment system for flow cytometry in Quercus[J]. Journal of Nanjing Forestry University:Natural Sciences Edition, 2015, 39(1):167-172.
[25] Bennett MD, Leitch IJ. Plant DNA C-value Database (release 6.0)[DB/OL]. 2012.[2017-01-01]. Http://data.kew.org/cvalues/.
计量
- 文章访问数: 1011
- HTML全文浏览量: 2
- PDF下载量: 1300
下载:
