Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2018, Vol. 53, ¹ 3, p. 531-538.
(how to cite this article)

doi: 10.15389/agrobiology.2018.3.531eng

UDC 633.18:631.527:581.143.6:575.2

Acknowledgments:
The authors would like to deeply thank colleagues from the Laboratory of Rice Breeding (Primorskii Research Institute of Agriculture) for the hybrid rice material provided.

Supported in part by Far East Program for Basic Research of the Far Eastern Branch RAS (grant ¹ 15-I-6-005)

 

 

NUCLEAR DNA CONTENT IN RICE (Oryza sativa L.) REGENERANTS DERIVED FROM ANTHER CULTURE in vitro

M.V. Ilyushko1, M.V. Skaptsov2, M.V. Romashova1

1Primorskii Research Institute of Agriculture, Federal Agency for Scientific Organizations, 30, ul. Volozhenina, pos.  Timityazevskii, Primorskii Krai, 692539 Russia, e-mail ilyushkoiris@mail.ru (✉ corresponding author);
2South Siberian Botanical Garden of Altai State University, 61, prosp. Lenina, Barnaul, 656049 Russia, e-mail mr.skaptsov@mail.ru

ORCID:
Ilyushko M.V. orcid.org/0000-0001-7042-8641
Skaptsov M.V. orcid.org/0000-0002-4884-0768
Romashova M.V. orcid.org/0000-0002-7426-8523

Received January 30, 2017

 

Rice is an important food crop grown in the south of the Russian Far East. Therefore, breeding new varieties with high harvest and crop quality is relevant. Anther in vitro culture is successfully applied in breeding programs in rice-growing countries, including Russia. In anther in vitro culture, flow cytometry is applicable to select haploid, dihaploid and polyploid regenerants. Cytological studies show genome variations from haploids to hexaploids in plant tissue in vitro culture, and also chromosome changes which result in aneuploidy or endopolyploidy leding to an inconstant nuclear DNA content. In the work, we followed the aims i) to evaluated nuclear DNA content by flow cytometry in an androgenic rice regenerant population, and ii) to estimate the applicability of the combination of two approaches, the anther in vitro culture technique and flow cytometry, in rice breeding. A total of 1099 regenerants from in vitro anther culture of a single F2 (UkrNIIS 3435 × Ukr 96) rice (Oryza sativa L. ssp. japonica Kato) hybrid plant were separated into four groups with regard to morphological features. Haploids were sterile plants with very small flowers, dihaploids were fertile plants, tetraploids were the plants with very few large seeds, an expressed keel and the ribbed floral scales. Also, there were the plants without seeds which flowers were normal in size but formed two or more sterile panicles. In the last group of the regenerants the plants died during early development. A total of 176 regeneranats were estimated by flow cytometry. It was revealed that nuclear DNA content varied greatly (Cv = 32 %) in the plants without seeds. This group seems to include plants with double set of chromosomes, triploids, tetraploids, and pentaploids. Additionally, in this group we found the regenerants with endopoyploidy since five of the plants had two nuclear DNA content peaks like those for haploids and diploids. In 23 plants nuclear DNA content approximated to dihaploid chromosome set and averaged 2.00 pg. Obviously, an aneuploidy characteristic of rice anther in vitro cultures could lead to aliquant changes in chromosome set in the regenerants, causing a loss of fertility. The dihaploid and tetraploid plants were low variable (Cv of 10.5 and 5.3 %) and had nuclear DNA content of 1.88 and 3.75 pg, respectively, whereas the haploids were high variable (Cv = 29 %) with an average nuclear DNA amount of 0.89 pg. Our findings indicate that flow cytometry, together with production index, may be applied to reveal tetraploid regenerants and to remove haploids in rice breeding. That allows avoiding ex vitro trials of unpromising regenerants.

Keywords: Oryza sativa L., anther culture in vitro, flow cytometry, regenerant, haploid, dihaploid, tetraploid.

 

Full article (Rus)

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REFERENCES

  1. Chaika A.K. V sbornike: Dal'nevostochnaya nauka — agropromyshlennomu proizvodstvu regiona [In: Far East science for agro-industrial production of the region]. Vladivostok, 2008: 37-54 (in Russ.).
  2. Sakhno A.L., Ilyushko M.V. Materialy Mezhvuzovskoi nauch.-prakt. konferentsii «Molodye uchenye — agropromyshlennomu kompleksu Dal'nego Vostoka». Ussuriisk, 2011, vyp. 11: 44-52 (in Russ.).
  3. Datta S.K. Androgenic haploids: factors controlling development and its application in crop improvement. Current Science, 2005, 89: 1870-1878.
  4. Dunwell J.M. Haploids in flowering plants: origins and exploitation. Plant Biotechnol. J., 2010, 8: 377-424 CrossRef
  5. Katalog sortov risa i ovoshchebakhchevykh kul'tur kubanskoi selektsii (spravochno-metodicheskoe izdanie) [Catalog of varieties of rice and vegetables and cabbage crops of the Kuban breeding (reference and methodical edition)]. Krasnodar, 2016 (in Russ.).
  6. Zmeeva V.N. Tendentsii izmenchivosti nekotorykh khozyaistvenno poleznykh priznakov v populyatsiyakh somaklonov i androgennykh digaploidov risa Oryza sativa L. Avtoreferat kandidatskoi dissertatsii [Trends in the variability of some economically useful traits in populations of somaclones and androgenic digaploid rice Oryza sativa L. PhD Thesis]. Vladivostok, 1995 (in Russ.).
  7. Ilyushko M.V. Materialy nauchno-prakticheskoi konferentsii «Rol' agrarnoi nauki v obespechenii prodovol'stvennoi bezopasnosti dal'nevostochnogo regiona: (k 40-letiyu Primorskogo NIISKH)» [Proc. Conf. on the role of agrarian science in ensuring food security in the Far Eastern region]. Vladivostok, 2016: 75-79 (in Russ.).
  8. Yamamoto T., Soeda Y., Nishikawa A., Hirohara H. A study of somaclonal variation for rice improvement induced by three kinds of anther-derived cell culture techniques. Plant Tissue Culture Letters, 1994, 11: 116-121 CrossRef
  9. Kucherenko L.A., Kharchenko P.N., Kovaleva E.N. Materialy Vsesoyuznoi konferentsii «Sostoyanie i perspektivy razvitiya sel'skokhozyaistvennoi biotekhnologii» [State of art and prospects of development of agricultural biotechnology]. Leningrad, 1986: 92-96 (in Russ.).
  10. Ilyushko M.V. Vestnik Rossiiskoi sel'skokhozyaistvennoi nauki, 2016, 5: 20-22 (in Russ.).
  11. Kucherenko L.A., Kharchenko P.N. Sel'skoe khozyaistvo za rubezhom, 1975, 11: 20-21 (in Russ.).
  12. Ochatt S.J. Flow cytometry in plant breeding. Cytometry, 2008, 73A: 581-598 CrossRef
  13. Murovec J., Bohanec B. Haploid and double haploids in plant breeding. In: Plant breeding. Rijeka, Croatia, 2012, Ch. 5: 87-106.
  14. Xu L., Najeeb U., Tang G.H. Haploid and doubled haploid technology. Adv. Bot. Res., 2007, 45: 181-216 CrossRef
  15. Li Y., Li H., Chen Z., Ji L.-X., Ye M.-X., Wang J., Wang L., An X.-M. Haploid plants from anther culture of poplar (Populus × beijingensis). Plant Cell Tiss. Org., 2013, 114: 39-48 CrossRef
  16. Sood S., Dwivedi S., Reddy T.V., Prasanna P.S., Sharma N. Improving androgenesis-mediated doubled haploid production efficiency of FCV tobacco (Nicotiana tabacum L.) through in vitro colchicines application. Plant Breeding, 2013, 132: 764-771 CrossRef
  17. Germana M.A. Gametic embryogenesis and haploid technology as valuable support to plant breeding. Plant Cell. Rep., 2011, 30: 839-857 CrossRef
  18. Goncharova Yu.K. Ispol'zovanie metoda kul'tury pyl'nikov v selektsii risa [The use of the anther culture in rice selection]. Krasnodar, 2012 (in Russ.).
  19. Gorbunova V.Yu. Androgenez in vitro u yarovoi myagkoi pshenitsy. Avtoreferat kandidatskoi dissertatsii [Androgenesis in vitro in spring soft wheat. PhD Thesis]. St. Petersburg, 2000 (in Russ.).
  20. Barow M., Jovtchev G. Endopolyploidy in plants and its analysis by flow cytometry. In: Flow cytometry with plant cell. J. Dolezel, J. Greilhuber, J. Suda (eds.). Wiley-VCH Verlag, Weinheim, 2007: 349-372 CrossRef
  21. Chu C. The N6 medium and its applications to anther culture of cereal crops. Proc. Simposium on Plant Tissue Culture. Peking, 1978: 43-50.
  22. Pfosser M., Amon A., Lelley T., Heberle-Bors E. Evaluation of sensitivity of flow cytometry in detecting aneuploidy in wheat using disomic and ditelosomic wheat-rye addition lines. Cytometry, 1995, 21(4): 387-393 CrossRef
  23. Skaptsov M.V., Smirnov S.V., Kutsev M.G., Shmakov A.I. Turczaninowia, 2016, 19(3): 120-122 (in Russ.).
  24. Skaptsov M.V., Smirnov S.V., Kutsev M.G. Turczaninowia, 2014, 17: 72-78 CrossRef (in Russ.).
  25. Skaptsov M.V., Lomonosova M.N., Kutsev M.G., Smirnov S.V., Shmakov A.I. The phenomenon of endopolyploidy in some species of the Chenopodioideae (Amaranthaceae). Botany Letters, 2017, 164(1): 47-53 CrossRef
  26. Barow M. Endopolyploidy in seed plants. BioEssays, 2006, 28: 271-281 CrossRef
  27. Bai C.K., Alverson W.S., Follansbee A., Waller M.D. New reports of nuclear DNA content for 407 vascular plant taxa from the United States. Annals of Botany, 2012, 110: 1623-1629 CrossRef
  28. Bennett M.D., Smit J.B. Nuclear DNA amounts in angiosperms. Phil. Trans. R. Soc. Lond. B, 1991, 334: 309-345 CrossRef
  29. Maletskii S.I., Yudanova S.S., Maletskaya E.I. Analysis of epigenomic and epiplastome variability in the haploid and dihaploid sugar beet (Beta vulgaris L.) plants. Sel’skokhozyaistvennaya biologiya [Agricultural Biology], 2015, 50: 579-589 CrossRef
  30. Goncharova Yu.K. Genetika, 2013, 49: 196-203 CrossRef (in Russ.).
  31. Luan L., Wang X., Long W.B., Liu Y.H., Tu S.B., Xiao X.Y., Kong F.L. A comparative cytogenetic study of the rice (Oryza sativa L.) autotetraploid restorers and hybrids. Genetika, 2009, 45: 1225-1233.
  32. Barker R.E., Kilgore J.A., Cook R.L., Garay A.E., Warnke S.E. Use of flow cytometry to determine ploidy level of ryegrass. Seed Sci. Technol., 2001, 29: 493-502.
  33. Smarda P., Bures P., Horova L., Foggi B., Rossi G. Genome size and GC content evolution of Festuca: Ancestral expansion and subsequent reduction. Annals of Botany, 2008, 101: 421-433 CrossRef
  34. Bennett M.D., Smith J.B. Nuclear DNA amounts in angiosperms. Philos. T. Roy. Soc. B, 1976, 274: 227-274 CrossRef

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