CHROMOSOME COUNT ON YOUNG ANTHER OF BANANA MALE BUD USING EZYMATIC MACERATION AND DAPI STAINING IN SLIDE PREPARATION

Fajarudin Ahmad, Yuyu Suryasari Poerba
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Abstract

Chromosome counting is the basis in describing the chromosomes number of organism that might useful for genetic study and classification. In banana studies, the root tip with a combination of non-fluorescent staining methods such as carmine or orcein and squash is the most common material for chromosome counting. In this study, we presented the usefulness of young anther of banana male bud with enzymatic maceration method for cell spreading and 4,6-diamino-2-phenyl-indole (DAPI) for staining agent to get a satisfying chromosomes image at metaphase for mitotic study of diploid and tetraploid bananas. The principle of this study is fixation using ethanol:acetic acid (3:1), enzymatic digestion, maceration and staining using DAPI. Our result showed that this method can provide well spread cells with intensely contrast of chromosomes images that satisfying for chromosome counting.

 

Keywords

Diploid, Fluorescent, Metaphase, Mitotic, Staining, Tetraploid

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References

Adeleke, M.T .V, Pillay, M. and Okoli, B.E., 2002. An improved method for examining meiotic chromosomes in Musa L. HortScience, 37(6), pp. 959–961.

Anamthawat-Jónsson, K., 2014. Preparation of chromosomes from plant leaf meristems for karyotype analysis and Preparation of chromosomes from plant leaf meristems for karyotype analysis and in situ hybridization. Method in Cell Science, 25, pp. 91–95.

Antonin, W. and Neumann, H., 2016. Chromosome condensation and decondensation during mitosis. Current Opinion in Cell Biology, 40, pp. 15–22.

Bakry, F. and Shepherd, K., 2014. Banana protocol chromosome count on banana root tip squashes. Fruits, 63, pp. 179–181.

Cheesman, E.E., 1947. Classification of the bananas: the genus Musa L. Kew Bulletin, 2, pp. 106–117.

Crang, R., Lyons-sobaski, S. and Wise, R., 2018. Plant Anatomy: A Concept-Based Approach to the Structure of Seed Plants. Springer Nature. Cham. pp. 123–154.

D’Hont, A., Paget-Goy, A., Escoute, J. and Carreel, F., 2000. The interspecific genome structure of cultivated banana, Musa spp. revealed by genomic DNA in situ hybridization. Theoretical and Applied Genetics, 100, pp. 177–183.

Doležel, J., Doleželová, M., Roux, N. and Van den Houwe, I., 1998. A novel method to prepare slides for high resolution chromosome studies in Musa spp. Infomusa, 7, pp. 3–4.

Doležel, J., Lysak, M.A., van Den Houwe, I., Dolezelova, M. and Roux, N., 1997. Use of flow cytometry for rapid ploidy determination in Musa species. Infomusa, 6, pp. 6–9.

Ferguson-Smith, M.A., 2002. Satellited Chromosome. In: Brener, S. and Miller, J.H. eds. Ensiclopedia of Genetics. pp. 1773–1774. Academic Press.

Heslop-Harrison, J., Osuji, J. and Hull, R., 1998. Fluorescent in situ hybridization of plant chromosomes: illuminating the Musa genome. In: INIBAP: Networking Banana and Plaintain: INIBAP Annual Report 1998. pp. 26–29. International Network for the Improvement of Banana and Plaintain, Montpellier, France.

Kanchanapoom, K. and Koarapatchaikul, K., 2012. In vitro

induction of tetraploid plants from callus cultures of diploid bananas (Musa acuminata, AA group) “Kluai Leb Mu Nang” and “Kluai Sa”. Euphytica, 183, pp. 111–117.

Kantama, L., Wijnker, E. and de Jong, H., 2017. Optimization of Cell Spreading and Image Quality for the Study of Chromosomes in Plant Tissues. In: Schmidt, A. ed. Plant Germline Development: Methods and Protocols. pp. 141–158. Humana Press. New York.

Lusinska, J., Majk, J., Betekhtin, A., Susek, K., Wolny, E. and Hasterk, R., 2018. Chromosome identification and reconstruction of evolutionary rearrangements in Brachypodium distachyon, B. stacei and B. hybridum. Annals of Botany, 122, pp. 445–459.

Mora-Bermúdez, F., Gerlich, D. and Ellenberg, J., 2007. Maximal chromosome compaction occurs by axial shortening in anaphase and depends on Aurora kinase. Nature Cell Biology, 9, pp. 822–831.

Ortiz, R., 2000. Understanding the Musa genome: An update. Acta Horticulturae, 340, pp. 157–168.

Osuji, J., Okoli, B.E. and Ortiz, R., 1996. An improved procedure for mitotic studies of of the Eumusa section of the genus Musa L. (Musaceae). Infomusa, 5, pp. 12–14.

Osuji, J., Okoli, B.E. and Edeoga, H.O., 2006. Karyotypes of the A and B Genomes of Musa L. Cytologia, 71, pp. 21–24.

Poerba, Y.S., Handayani, T. and Witjaksono, 2017. Characterization of tetraploid Pisang Rejang induced by oryzalin. Berita Biologi, 16, pp. 85–93.

Rekha, A. and Hiremath, S.C., 2008. Chromosome studies and karyotype analysis of some triploid banana (Musa species) cultivars of AAA genomic group. Journal of Horticultal Science, 3, pp. 30–34.

Roux, N., Toloza, A., Radecki, Z., Zapata-Arias, F.J. and Dolezel, J., 2003. Rapid detection of aneuploidy in Musa using flow cytometry. Plant Cell Reports, 21, pp. 483–490.

Sapre, A.B. and Barve, S., 1983. Somatic chromosomes from intercalary. Cytologia, 48, 539–541.

Sheikh, S. and Kondo, K., 1995. Differential Staining with Orcein, Giemsa, CMA and DAPI for Comparative Chromosome Study of 12 Species of Australian Drosera (Droseraceae). American Journal of Botany, 82, pp. 1278–1286.

Shepherd, K., 1999. Cytogenetics of the genus Musa. International Network for the Improvement of Banana and Plantain. Montpellier, France. pp. 9–14.

Schneider, C.A., Rasband, W.S. and Eliceiri, K.W., 2012. NIH Image to ImageJ: 25 years of image analysis. Nature Methods, 9(7), pp. 671–675.

Wong, C., Kiew, R., Argent, G., Set, O., Lee, S.K. and Gan, Y.Y., 2002. Assessment of the validity of the sections in Musa (Musaceae) using AFLP. Annals of Botany, 90, pp. 231–238.


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