Welcome to Acta Agronomica Sinica，

Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (10): 1794-1800.doi: 10.3724/SP.J.1006.2011.01794

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Analyzing Synchronization Process of Mitosis Cells in Wheat Root-tip with Flow Karyotype

GUO Dong-Wei1,ZHANG Ren-He1,LI Chun-Lian1,CHEN Yao-Feng1,MIN Dong-Hong1,*,CHEN Ming2,LI Lian-Cheng2,XU Zhao-Shi2,MA You-Zhi2   

  1. 1 Agronomy College, Northwest A & F University, Yangling 712100, China; 2 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2011-01-13 Revised:2011-06-25 Online:2011-10-12 Published:2011-07-28
  • Contact: 闵东红, E-mail: mdh2493@nwsuaf.edu.cn E-mail:gdwei1973@126.com

PDF

1789

Abstract: Genomic sequencing of wheat can be simplified through constructing chromosome specific library. Synchronization of cells is propitious to high mitosis index (MI), which results in the accumulation of metaphase chromosomes and a successful flow sorting. In the present study, we used a double-blocked strategy in the synchronization treatment of wheat (Triticum aestivum L. cv. Chinese Spring) root-tip cells with 50 µmol L-1 hydroxyurea (HU) and 100 µmol L-1 trifluralin. The effects of treatments were evaluated by analyzing the dynamic proportions of cells at different stages of cell cycle. The optimal hours of treatments were 10 h for HU block, 7 h for recovery, and 4 h for trifluralin block. According to this synchronization scheme, the maximum MI was 70.1%. Additional ice water (0°C) depression was helpful for decreasing chromosome debris and rising resolution of the flow karyotype, but had no effect on the accumulation of metaphase chromosomes in root-tip cells.

Key words: Wheat, Mitosis, Synchronization, Flow karyotype, Flow sorting

[1]Gill B S, Appels R, Botha-Oberholster A M, Robin Buell C, Bennetzen J L, Chalhoub B, Chumley F, Dvoák J, Iwanaga M, Keller B, Li W, McCombie W R, Ogihara Y, Quetier F, Sasaki T. A workshop report on wheat genome sequencing: international genome research on wheat consortium. Genetics, 2004, 168: 1087–1096
[2]Vrana J, Kubalakova M, Simkova M, Cihalikovaa J, Lysáka M A,Dolezela J. Flow sorting of mitotic chromosomes in common wheat (Triticum aestivum L.). Genetics, 2000, 156: 2033–2041
[3]Paux E, Sourdille P, Salse J, Saintenac C, Choulet F, Leroy P,Korol A, Michalak M, Kianian S, Spielmeyer W, Lagudah E,Somers D, Kilian A, Alaux M, Vautrin S, Berges H, Eversole K,Appels R, Safar J, Simkova H, Dolezel J, Bernard M, Feuillet C. A physical map of the 1-gigabase bread wheat chromosome 3B.Science, 2008, 322: 101–104
[4]Janda J, Bartos J, Safar J, Kubalakova M, Valarik M, Cihalikova J, Simkova H, Caboche M, Sourdille P, Benard M, Chalhoub B, Dolezel J. Construction of a subgenomic BAC library specific for chromosomes 1D, 4D and 6D of hexaploid wheat. Theor Appl Genet, 2004, 109: 1337–1345
[5]Safar J, Simkova H, Kubalakova M, Cihalikova J, Suchankova P,Bartos J, Dolezel J. Development of chromosome-specific BAC resources for genomics of bread wheat. Cytogenetic & Genome Res, 2010, 129: 211–223
[6]Lee J H, Arumuganathan K, Yen Y, Kaeppler S, Kaeppler H,Baenziger P S. Root tip cell cycle synchronization and metaphase chromosome isolation suitable for flow sorting in common wheat (Triticum aestivum L.). Genome, 1997, 40: 633–638
[7]Dolezel J, Kubalakova M, Paux E, Bartos J, Feuillet C. Chromosome-based genomics in the cereals. Chromosome Res, 2007, 15:51–66 1800 作 物 学 报 第 37 卷
[8]Kubalakova M, Kubalakova P, Suchankova P, Cihalikova J, Bartos J, Lucretti S, Wantanabe N, Kianian J, Dolezel J. Chromosome sorting in tetraploid wheat and its potential for genome analysis. Genetics, 2005, 170: 823–829
[9]Pan W H, Houben A and Schlegel R. Highly effective cell synchronization in plant roots by hydroxyurea and amiprophos-methyl or colchicines. Genome, 1993, 36: 387–390
[10]Suchankova P, Simkova, H, Svensson J T, Hribova E, Condamine P, Bhat P R, Bartos J, Safar J, Close T J, Dole?el J. Production of microgram amounts of DNA from flow-sorted plant chromosomes. Chromosome Res, 2007, 15: 4–5
[11]Clain E, Brulfert A. Hydroxyurea-induced mitotic synchronization in Allium sativum root meristems. Planta, 1980, 150: 26–31
[12]Bouget F V, Moulager M, Corellou F. Circadian regulation of cell division. In: Verma D P S, Hong Z, eds. Cell Division Control in Plants. Verlag Berlin Heideberg: Springer, 2007. pp 3–12
[13]Guo D W, Ma Y Z, Li L C, Chen Y F. Flow sorting of wheat chromosome arms from the ditelosomic 7BL. Plant Mol Biol Rep, 2006, 24: 23–31
[14]Luo M, Ma Y, You F M, Anderson O D, Kopecky D, Simkova H,Safar J, Dole?el J, Gill B, McGuire P, Dvorak J. Feasibility of physical map construction from fingerprinted bacterial artificial chromosome libraries of polyploid plant species. BMC Genomics,2010, 11: 122
[15]Mayer K F X, Taudien S, Martis M, Šimková H, Suchánková P,Gundlach H, Wicker T, Petzold A, Felder M, Steuernagel B,Scholz U, Graner A, Platzer M, Dole?el J, Stein N. Gene content and virtual gene order of barley chromosome 1H. Plant Physiol,2009, 151: 496–505
[16]Simkova H, Safar J, Kubalakova M, Suchankova P, Cihalikova J,Robert-Quatre H, Azhaguvel P, Weng Y, Peng J, Lapitan Nora L V, Ma Y, You F M, Luo M, Bartos J, Dolezel J. BAC libraries from wheat chromosome 7D: efficient tool for positional cloning of aphid resistance genes. J Biomedicine & Biotechnol, 2011, doi: 10.1155/2011/302543
[17]Kovarova P, Navratilova A, Macas J, Dolezel J. Chromosome analysis and sorting in Vicia sativa using flow cytometry. Biol Plant, 2007, 51: 43–48
[18]Nompumelelo H M, Walter D, Margit M, Spencer M, Carl C,Rachael H, Jeroen N, Simon S, James A H M. D-type cyclins activate division in the root apex to promote seed germination in Arabidopsis. Proc Natl Acad Sci USA, 2005, 102: 15694–15699
[19]Huberman J A. New views of the biochemistry of eukaryotic DNA replication revealed by aphidicolin, an unusual inhibitor of DNA polymerase alpha. Cell, 1981, 23: 647–648
[20]Simkova H, Safar J, Suchankova P, Kovarova P, Bartoa J, Kubalakova M, Janda J, Cihalikova J, Mago R, Lelley T, Dolezel J.Novel resource for genomics of Triticeae: BAC library specific for the short arm of rye (Secale cereale L.) chromosome 1R (1RS). BMC Genomics, 2008, 9: 237
[1] Mao Jia-Qi, Huang Peng-Yu, Zhao Jia-Jia, Zheng Xing-Wei, Wu Bang-Bang, Hao Yu-Qiong, Qu Fei, Liu Cheng, Ma Peng-Tao, Zheng Jun. Evaluation of powdery mildew resistance in wheat cultivars and molecular detection of resistance genes in Shanxi province, China [J]. Acta Agronomica Sinica, 2026, 52(6): 1669-1681.
[2] Hu Chuan, Zhao Kai-Nan, Huang Xiu-Li, Wu Jin-Zhi, Ren Kai-Ming, Wang He-Zheng, Fu Guo-Zhan, Huang Ming, Li You-Jun. Effects of tillage methods and nitrogen rates on yield and quality of dryland wheat under one-off irrigation [J]. Acta Agronomica Sinica, 2026, 52(6): 1830-1846.
[3] Chen Xue-Yan, He Hua-Chuan, Li Zheng-Jia, Dong Xin-Pan, Li Ou-Qi, Liu Xiao-Yun, Li Dan-Ping, Chen Zhi-Wei, Liu Guo-Xia, Lyu Sheng-Yuan, Wu Yin-Ying, Zhao Zhen-Dong, Cao Xin-You, Wan He-Ping. Dynamic changes in root organic acid secretion and its transcriptional regulatory mechanisms in ‘Jimai 60’ seedlings under combined salinity-alkalinity stress in hydroponics [J]. Acta Agronomica Sinica, 2026, 52(6): 1859-1875.
[4] Gao Pei-Yang, Li Jin-Xuan, Dong Yu-Kui, Shi Yu, Zhang Zhen, Zhang Yong-Li. Response of wheat tillering and spike formation to nitrogen rate under supplementary irrigation based on soil moisture content [J]. Acta Agronomica Sinica, 2026, 52(6): 1847-1858.
[5] Zhang Xian-Feng, Guo Li-Jian, Li Kang-Chun, Kong Bin-Xue, Liu Yu-Fang, Che Zhuo, Yang De-Long. Identification of the ABHD6 gene family and development of functional markers for grain weight in wheat [J]. Acta Agronomica Sinica, 2026, 52(6): 1711-1727.
[6] Zhai Sheng-Nan, Cao Xin-You, Li Hao-Sheng, Li Ji-Hu, Li Fa-Ji, Liu Jin-Dong, Xia Xian-Chun, Lyu Ying-Ying, Ma Rui-Feng, Wang Ying, Geng Hong-Wei, Liu Jian-Jun. Analysis of the genetic effects of allelic variation at the Pod-A1, Pod-D1, and Pod-2D loci on peroxidase activity in wheat grains [J]. Acta Agronomica Sinica, 2026, 52(6): 1593-1603.
[7] Xi Qian-Hui, Xu Zi-Yuan, Liu Meng-Meng, Wang Hong-Yi, Lang Kai-Lin, Jing Zhen-Hai, Chen Feng, Zhao Lei. Genome-wide association study and candidate gene prediction of grain copper content in wheat [J]. Acta Agronomica Sinica, 2026, 52(6): 1604-1617.
[8] Wang Zhuang-Zhuang, Wu Zi-Jun, Zhang Yong-Xin, Zhang Xin-Yuan, Yuan Li-Xue, Chen Ru-Xue, Liu Shi-Ju, Duan Jian-Zhao, Feng Wei, Wang Tong-Chao, Wang Yong-Hua. Optimized water-nitrogen synergy enhances winter wheat yield and nitrogen use efficiency in clay-loam fluvo-aquic soils of southeastern Henan, China [J]. Acta Agronomica Sinica, 2026, 52(5): 1501-1521.
[9] He Wan-Long, Geng Hong-Wei, Zhang Fei-Fei, Mikereayi·Ababaikere , Luo Zi-Yang, Li Peng-Cheng, Zhou Zhao-Yu, Cheng Yu-Kun. Development of a deep learning-based image recognition system for major wheat diseases [J]. Acta Agronomica Sinica, 2026, 52(5): 1401-1417.
[10] Zhang Zhen, Feng Lian-Jie, Shi Yu, Yu Zhen-Wen, Zhang Yong-Li. Yield formation of wheat with different ear types under water-saving supplementary irrigation conditions [J]. Acta Agronomica Sinica, 2026, 52(5): 1522-1535.
[11] Hou Si-Yu, Wang Guo-Cui, Wei Jin-Gui, Xie Wei-Xin, Yin Wen, Fan Zhi-Long, Chai Qiang, Hu Fa-Long. Effects of green manure combined with chemical nitrogen fertilizer on dry matter accumulation and yield formation of wheat in arid irrigation areas of northwestern China [J]. Acta Agronomica Sinica, 2026, 52(4): 1208-1219.
[12] Shang Yun-Qiu, Zhao Zhu, Chen Huan, Ding Yong-Gang, Qiao Yu-Qiang, Li Wei, Zhang Xiang-Qian, Cao Cheng-Fu, Du Shi-Zhou. Effects of long-term tillage practices on grain-filling and yield formation in rain-fed wheat [J]. Acta Agronomica Sinica, 2026, 52(4): 1236-1250.
[13] Qiao Yu-Xin, Li Cheng-Yue, Kang Xiao-Yu, Zhang Xin-Qi, Jia Shao-Hui, Liu Qian, Cao Ya-Li, Shi Xin-Rui, Hao Xing-Yu, Li Ping. Study on the effects of long-term no-tillage straw mulching on wheat yield improvement in dryland areas based on the APSIM model [J]. Acta Agronomica Sinica, 2026, 52(4): 1181-1192.
[14] Li Can, Zhang Xi-Wei, Zhu Bo-Tao, Zhang Pei-Pei. Functional characterization of wheat GSK kinase TaSK41 and screening for interacting proteins [J]. Acta Agronomica Sinica, 2026, 52(3): 677-687.
[15] Hou Jie, Fu Duo-Duo, Wu Hai-Feng, Hao Yu-Qiong, Zheng Xing-Wei, Wu Bang-Bang, Zhou Kai, Li Xiao-Hua, Zheng Jun, Zhao Jia-Jia. Chromosome diversity and its effects in wheat landraces from Shanxi province, China [J]. Acta Agronomica Sinica, 2026, 52(3): 746-763.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd