[1]马秋月,李淑顺,马 骧,等.基于流式细胞技术的两种槭属植物基因组大小测定[J].南京林业大学学报(自然科学版),2018,42(05):201-205.[doi:10.3969/j.issn.1000-2006.201711030]
 MA Qiuyue,LI Shushun,MA Xiang,et al.Estimation of genome size of two Acer Linn. by flow cytometry[J].Journal of Nanjing Forestry University(Natural Science Edition),2018,42(05):201-205.[doi:10.3969/j.issn.1000-2006.201711030]
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基于流式细胞技术的两种槭属植物基因组大小测定
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《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

卷:
42
期数:
2018年05期
页码:
201-205
栏目:
研究简报
出版日期:
2018-09-15

文章信息/Info

Title:
Estimation of genome size of two Acer Linn. by flow cytometry
文章编号:
1000-2006(2018)05-0201-05
作者:
马秋月1李淑顺1马 骧2闻 婧1朱 璐1唐 玲1黎 瑞1郭 伟3李倩中1*
1.江苏农业科学院休闲农业研究所,江苏 南京 210014; 2.金坛区农林局林业站,江苏 常州 213200; 3.江苏省农业科学院种质资源与生物技术研究所,江苏 南京 210014
Author(s):
MA Qiuyue1 LI Shushun1 MA Xiang2 WEN Jing1 ZHU Lu1 TANG Ling1 LI Rui1 GUO Wei3 LI Qianzhong1*
1.Institute of Leisure Agriculture in Jiangsu Academy of Agricultural Sciences, Nanjing 210014,China; 2. Jintan Bureau of Agricultural and Forestry, Changzhou 213200, China; 3. Institute of Germplasm Resources and Biotechnology, Nanjing 210014, China
关键词:
槭属 鸡爪槭 元宝枫 基因组大小 流式细胞术
Keywords:
Acer Linn. A. palmatum A. truncatum genome size flow cytometry
分类号:
S687.9; S722
DOI:
10.3969/j.issn.1000-2006.201711030
文献标志码:
A
摘要:
【目的】槭属植物具有重要的园林观赏价值和药用价值,开发应用前景广阔,但该属植物分子生物学及基因组学研究还相对匮乏,此次研究利用流式细胞仪技术对该属中鸡爪槭和元宝枫基因组大小进行测定,为后期槭属植物的细胞遗传学和基因组学研究奠定基础。【方法】以鸡爪槭(Acer palmatum)和元宝枫(A. truncatum)的嫩叶为材料,首次采用LB01解离液和碘化丙啶(PI)荧光染料,利用基因组大小已知的玉米(Zea mays L.)CE-777作为内标,建立适合于该两种材料的流式细胞术基因组大小测定方法。【结果】LB01解离液对测定样品与内标解离效果均较好,粒子清晰集中,无重叠峰且区分度良好,最后测得鸡爪槭的基因组1C含量为0.840 4 pg,元宝枫的基因组1C含量为0.756 0 pg。【结论】基于流式细胞仪技术,选取合适的样品处理方法,可成功测定鸡爪槭和元宝枫的基因组大小。该结果和方法可丰富槭属植物的基因库,为揭示该属植物的物种起源和进化提供重要依据,也为其后期的基因组测序工作奠定了重要基础。
Abstract:
【Objective】Acer plants have potential value as ornamental plants and as a source of medicinal compounds. However, their use is hindered by a lack of molecular biology and genomic data. Presently, flow cytometry was used to examine Acer palmatum Thunb. and A. truncatum Bunge genome sizes, to provide foundational data for cytogenetics and genomics studies of Acer Linn.【Method】Fresh leaves of A. palmatum Thunb and A. truncatum were collected. LB01 and PI were used as isolation buffer. Zea mays L. CE-777 was used as the internal standard for the flow cytometry analyses. 【Result】Samples and the internal standard were better dissociated by the LB01 solution, with particles clearly distinguished and concentrated without overlapping peaks. The genome size of A. palmatum and A. truncatum was 0.840 4 and 0.756 0 pg, respectively.【Conclusion】Optimized sample processing and flow cytometry permitted the determinations of the genome sizes of A. palmatum Thunb and A. truncatum Bunge. The optimized processing and results enrich the Acer gene database and provide foundational data for investigations of origin and evolution of species, and genome sequencing.

参考文献/References:

[1] 方文培. 中国植物志:46卷[M]. 北京:科学出版社,1981.
[2] HOVANET M V, DOCIU N, DINU M, et al. A comparative physico-chemical analysis of Acer platanoides and Acer pseudoplatanus seed oils [J]. Revista de Chimie-Bucharest-Original Edition, 2015, 66(7):987-991.
[3] 王性炎. 中国元宝枫[M]. 成都:四川民族出版社,2003.
[4] 徐廷志. 槭属的系统演化与地理分布[J]. 植物分类与资源学报, 1998, 20(4):383-393.
XU T Z. The systematic evolution and distribution of the genus Acer [J]. Plant Diversity, 1998, 20(4):383-393.
[5] SWIFT H. The constancy of desoxyribose nucleic acid in plant nuclei [J]. Proceedings of the National Academy of Sciences of the United States of America, 1950, 36(11):643-654. DOI: 10.1073/pnas.36.11.643.
[6] LARGIA M J V, SHILPHA J, POTHIRAJ G, et al. Analysis of nuclear DNA content, genetic stability, Bacoside A quantity and antioxidant potential of long term in vitro grown germplasm lines of Bacopa monnieri(L.)[J]. Plant Cell Tissue & Organ Culture, 2015, 120(1):399-406. DOI:10.1007/s11240-014-0602-5.
[7] LEVINE R F, JR B P, HAZZARD K C, et al. Flow cytometric analysis of megakaryocyte ploidy: comparison with Feulgen microdensitometry and discovery that 8N is the predominant ploidy class in guinea pig and monkey marrow [J]. Blood, 1980, 56(2):210-217.
[8] BAUMGARTH N, ROEDERER M. A practical approach to multicolor flow cytometry for immunophenotyping [J]. Journal of Immunological Methods, 2000, 243(1-2):77-97.
[9] MARCO F, MARCO M, WARD J A, et al. An evaluation of the PacBio RS platform for sequencing and de novo assembly of a chloroplast genome [J]. Bmc Genomics, 2013, 14(1):670-670. DOI: 10.1186/1471-2164-14-670.
[10] BENNETT M D, LEITCH I J. Nuclear DNA amounts in angiosperms: targets, trends and tomorrow [J]. Annals of Botany, 2011, 107(3): 467-590. DOI: 10.1093/aob/mcq258.
[11] 张琳琳, 曹博, 白成科. 应用流式细胞术测定药用植物黄芩基因组大小[J]. 中国农学通报, 2013, 29(25):130-135.
ZHANG L L, CAO B, BAI C K. The detection of genome size of medicinal plant Scutellaria baicalensis georigi by flow cytometry [J]. Chinese Agricultural Science Bulletin, 2013, 29(25):130-135.
[12] 张靖靖. 紫草基因组大小的测定研究[D]. 南京:南京大学, 2013.
ZHANG J J. Study on the measuration of genome size of Lithospermum erythrorhizon [D]. Nanjing: Nanjing University, 2013.
[13] 邓果特, 刘清波, 蒋建雄,等. 五节芒基因组大小测定[J]. 植物遗传资源学报, 2013, 14(2):339-341. DOI:10.3969/j.issn.1672-1810.2013.02.024.
DENG G T, LIU Q B, JIANG J X, et al. Estimation of genome size of Miscanthus floridulus [J]. Journal of Plant Genetic Resource, 2013, 14(2):339-341.
[14] 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. DOI:10.3969/j.issn.1000-2006.2015.01.030.
WEI G M, FANG Y M. Establishment and optimization of an experiment system for flow cytometry in Quercus [J]. Journal of Nanjing Forsetry University(Natural Sciences Edition), 2015, 39(1):167-172.
[15] SILJAK-YAKOVLEV S, PUSTAHIJA F, SOLIC E M, et al. Towards a genome size and chromosome number database of balkan flora: C-values in 343 taxa with novel values for 242 [J]. Journal of Computational & Theoretical Nanoscience, 2010, 3(2):190-213. DOI:10.1166/asl.2010.1115.
[16] MARTIN A, LYSAK, JAROSLAV DOLEZEL. Estimation of nuclear DNA content in Sesleria(Poaceae)[J]. Caryologia, 1998, 51(2):123-132.
[17] 周翔宇, 林峰, 徐莉, 等. 基于流式细胞术的5种鼠尾草基因组C值测定[J].南方农业学报, 2017, 48(6):960-965. DOI:10.3969/j.issn.2095-1191.2017.06.03.
ZHOU X Y, LIN F, XU L, et al. Measurement of genomic C value of five Salvia species by flow cytometry[J], 2017, 48(6):960-965.
[18] 林峰, 周翔宇, 徐莉, 等. 几种鼠尾草属植物基因组C值测定[J]. 农业生物技术学报, 2017, 25(10): 1622-1628. DOI:10.3969/j.issn.1674-7968.2017.10.007.
LIN F, ZHOU X Y, XU L, et al. Estimation of genomic C value in several species of Salvia [J]. Journal of Southern Agriculture, 2017, 25(10): 1622-1628.
[19] 吴丽萍, 唐岩, 李颖岳,等. 枣和酸枣基因组大小测定[J]. 北京林业大学学报, 2013, 35(3):77-83. DOI:10.13332/j.1000-1522.2013.03.012.
WU L P, TANG Y, LI Y Y, et al. Estimation of genome size of Ziziphus jujuba and Z. acdiojujuba[J]. Journal of Beijing Forestry University, 2013, 35(3):77-83.
[20] 周知里, 熊智, 马焕成, 等. 两种木棉科植物基因组大小测定[J]. 西部林业科学, 2014(2):97-101. DOI: 10.16473/j.cnki.xblykx1972.2014.02.007.
ZHOU Z L, XIONG Z, MA H C, et al. Genome size of two bombacaceae plants [J]. Journal of West China Forsetry Science, 2014(2):97-101.
[21] 陈丙义, 李金凤, 霍恒志,等. 6种野生草莓基因组大小估算[J]. 果树学报, 2015(1):51-56.
CHEN B Y, LI J F, HUO H Z, et al. Estimation of genome size in six wild strawberry species [J]. Journal of Fruit Science, 2015(1):51-56. DOI: 10.13925/j.cnki.gsxb.20140169.
[22] 李祯, 伊贤贵, 顾宇,等. 山樱花基因组大小的测定[J]. 南京林业大学学报(自然科学版), 2014,38(s1):17-19. DOI: 10.3969/j issn.1000-2006. 2014.S1.004.
LI Z, YIN X G, GU Y, et al. The genome size determination of Cerasus serrulata [J]. Journal of Nanjing Forsetry University(Natural Sciences Edition), 2014,38(s1):17-19.
[23] LOUREIRO J, RODRIGUEZ E, DOLEEL J, et al. Two new nuclear isolation buffers for plant DNA flow cytometry: a test with 37 species [J]. Annals of Botany, 2007, 100(4):875-888. DOI:10.1093/aob/mcm152.
[24] TAYLOR, RANDOLPH W. A morphological and cytological study of reproduction in the genus Acer [J]. Southeast Asian Journal of Tropical Medicine & Public Health, 2002, 33 suppl 2:115-123.
[25] 王性炎, 王姝清. 神经酸新资源——元宝枫油[J]. 中国油脂, 2005, 30(9):62-64.
[26] DOLEZEL J, BARTOS J, VOGLMAYR H, et al. Nuclear DNA content and genome size of trout and human [J]. Cytometry Part A, 2003, 51(2):127-128. DOI: 10.1002/cyto.a.10013.
[27] GREILHUBER J, DOLEZEL J, LYSáK M A, et al. The origin, evolution and proposed stabilization of the terms ‘genome size' and ‘C-value' to describe nuclear DNA contents[J]. Annals of Botany, 2005, 95(1):255-260. DOI: 10.1093/aob/mci019.
[28] OLSZEWSKA M J, OSIECKA R. The relationship between 2 C DNA content, systematic position, and the level of nuclear DNA endoreplication during differentiation of root parenchyma in some dicotyledonous shrubs and trees: comparison with herbaceous species [J]. Biochemie Und Physiologie Der Pflanzen, 1984, 179(8):641-657.
[29] BANCHEVA S, GREILHUBER J. Genome size in Bulgarian centaurea s.l.(Asteraceae)[J]. Plant Systematics & Evolution, 2006, 257(1/2):95-117. DOI:10.1007/s00606-005-0384-7.
[30] KUO L Y, HUANG Y J, CHANG J, et al. Evaluating the spore genome sizes of ferns and lycophytes: a flow cytometry approach [J]. New Phytologist, 2017, 213(4):1974. DOI: 10.1111/nph.14291.
[31] LAMATSCH D K, SCHMID M, SCHARTL M, et al. Noninvasive determination of genome size and ploidy level in fishes by flow cytometry: detection of triploid Poecilia formosa[J]. Cytometry Part A, 2015, 39(2):91-95. DOI: 10.1023/A:1022504602101.
[32] PAULE J, GREGOR T, SCHMIDT M, et al. Chromosome numbers of the flora of Germany: a new online database of georeferenced chromosome counts and flow cytometric ploidy estimates[J]. Plant Systematics & Evolution, 2016, 303(8):1-7. DOI:10.1007/s00606-016-1362-y.
[33] MISSET M T, GOURRET J P. Flow cytometric analysis of the different ploidy levels observed in the genus Ulex L. Faboideae-Genisteae in Brittany(France)[J]. Plant Biology, 2015, 109(1):72-79.
[34] PUTTICK M N, JAMES C, DONOGHUE P C J. Size is not everything: rates of genome size evolution, not C L. Faboideae-Genisteae in Brittany(France)[J]. Plant Biology, 2015, 109(1):72-79.
[34] PUTTICK M N, JAMES C, DONOGHUE P C J. Size is not everything: rates of genome size evolution, not lue, correlate with speciation in angiosperms [J]. Proceedings of the Royal Society B Biological Sciences, 2015, 282(1820):20152289. DOI:10.1098/rspb.2015.2289.

备注/Memo

备注/Memo:
收稿日期:2017-11-09 修回日期:2018-05-22 基金项目:农业科技自主创新基金项目(CX(17)1004); 国家自然科学基金项目(31700628) 第一作者:马秋月(yue.870808@163.com),助理研究员,博士。*通信作者:李倩中(546707668@qq.com),研究员。
更新日期/Last Update: 2018-09-15