Visn. Hark. nac. agrar. univ., Ser. Biol., 2019, Issue 2 (47), pp. 92-100


https://doi.org/10.35550/vbio2019.02.092




INFLUENCE OF 6-BENZYLAMINOPURINE ON INTENSITY OF CALLUSOGENESIS AND ORGANOGENESIS OF LINUM USITATISSIMUM L. UNDER IN VITRO CONDITIONS


S. V. Mishchenko

Institute of Bast Crops of National Academy of Agrarian Science of Ukraine

(Hlukhiv, Sumy region, Ukraine)


Linum usitatissimum L. convar. elongatum (Hlinum variety) is mostly capable to form the callus on the hypocotyl segments provided that it is cultivated on a Murashige and Skoog medium supplemented with 30 g/L sucrose, a 16 h photoperiod, a 60–80% relative humidity, an 22-24°C air temperature, and under the influence of only auxins (i), only cytokinins (ii) and combinations of auxins and cytokinins (iii) of exogenous origin. The highest frequency of organogenesis was in the variant with 1,0 mg/L 6-benzylaminopurine (BAP), 0.3 mg/L kinetine (cytokinins only), a combination of 1-naphthylacetic acid (NAA) and BAP, indole-3-acetic acid (IAA) and BAP. Studies with a large variation range of BAP concentrations indicate that in the presence of auxin NAA (0.05 mg/L in all variants) the efficiency of callus formation and shoots regeneration was higher. However, the use of only BAP can be quite a distinct technique for the induction of callus and organogenesis. For the studied species in general and this genotype in particular the synthesis of auxins of endogenous origin in the presence of cytokinins of exogenous origin is sufficient for organogenesis. Increasing of the concentration of BAP up to 3.0 mg/L inhibited the callus formation and shoots regeneration. The optimal concentrations of phytohormones can be expressed by inequality 1.0 ≤ BAP ≤ 1.75, and when 0.05 mg/L of NAA is added to the medium, inequality 0.5 ≤ BAP ≤ 2.0. The greatest efficiency of callus formation and organogenesis was found in variants 1.5 mg/L BAP (i), 1.25 mg/L BAP and 0.05 mg/L NAA (ii). Reaction of L. usitatissimum L. convar. elongatum on the effect of phytohormones compared to L. usitatissimum L. convar. mediterraneum was different.


Key words: Linum usitatissimum, in vitro, phytohormones, 6-benzylaminopurine, callus, organogenesis

 


REFERENCES


1. Polyakov A.V. 2000. Biotechnology in flax breeding. Tver : 180 p.
 
2. Soroka A.I. 2010. Peculiarities of donor plant preparation and flax anther cultivation in vitro for haploid plant production. Visnyk Zaporizkogo Nationalnogo Universytetu. Biologichni Nauky. 2 : 13-18.
 
3. Chen Y., Dribnenki P. 2002. Effect of genotype and medium composition on flax Linum usitatissimum L. anther culture. Plant Cell Rep. 21 (3) : 204-207. Doi: 10.1007/s00299-002-0500-x
https://doi.org/10.1007/s00299-002-0500-x
 
4. Blinstrubienė A., Burbulis N., Kuprienė R. 2009. Regeneration of adventitious shoots of linseed (Linum usitatissimum L.) from hypocotyl explants. Zemdirbyste-Agriculture. 96 (3) : 168-175.
 
5. Blinstrubienė A., Burbulis N., Kuprienė R. 2011. Effect of genotype and medium composition on linseed (Linum usitatissimum) ovary culture. Biologia. 66 (3) : 465-469. Doi: 10.2478/s11756-011-0028-z
https://doi.org/10.2478/s11756-011-0028-z
 
6. Blinstrubienė A., Burbulis N., Masienė R. 2017. Genotypic and exogenous factors affecting linseed ovary culture. Zemdirbyste-Agriculture. 2017. 104 (3) : 243-248. Doi: 10.13080/z a.2017.104.031
https://doi.org/10.13080/z-a.2017.104.031
 
7. Burbulis N., Blinstrubienė A., Sliesaravičius A., Venskutonienė E. 2005. Influence of genotype, growth regulators, sucrose level and preconditioning of donor plants on flax (Linum usitatissimum L.) anther culture. Acta Biologica Hungarica. 56 (3-4) : 323-331. o i: 10.1556/ABiol.56.2005.3 4.15
https://doi.org/10.1556/ABiol.56.2005.3-4.15
 
8. Burbulis N., Blinstrubienė A. 2011. Genotypic and exogenous factors affecting linseed (Linum usitatissimum L.) anther culture. J. Food Agricult. Environ. 9 (3-4) : 364-367. Doi: 10.1234/4.2011.2285
 
9. Burbulis N., Blinstrubienė A., Masienė R., Jonytienė V. 2012. Influence of genotype, growth regulators and sucrose concentration on linseed (Linum usitatissimum L.) anther culture. J. Food, Agricult. Environ. 10 (3-4) : 764-767. Doi: 10.1234/4.2012.3509
 
10. Diederrichsen A., Richards K. 2003. Taxonomy and germplasm conservation. In: Flax: The genus Linum (Ed. by A.D. Muir and N.D. Westcott). Boca Raton, USA, CRC Press : 39-42.
 
11. Evtimova M., Vlahova M., Atanassov A. 2005. Flax improvement by biotechnology means. Journal of Natural Fibers. 2 (2) : 17-34. Doi: 10.1300/J395v02n02_02
https://doi.org/10.1300/J395v02n02_02
 
12. Janowicz J., Niemann J., Wojciechowski A. 2012. The effect of growth regulators on the regeneration ability of flax (Linum usitatissimum L.) hypocotyl explants in in vitro culture. BioTechnologia. 93 (2) : 135-138. Doi: 10.5114/bta.2012.46578
https://doi.org/10.5114/bta.2012.46578
 
13. Millam S., Davidson D., Powell W. 1992. The use of flax (Linum usitatissimum) as a model system for studies on organogenesis in vitro: the effect of different carbohydrates. Plant Cell Tiss. Organ. Cult. 28 (2) : 163-166. Doi : 10.1007/BF00055512
https://doi.org/10.1007/BF00055512
 
14. Millam S., Obert B., Pretová A. 2005. Plant cell and biotechnology studies in Linum usitatissimum - a review. Plant Cell Tiss. Organ. Cult. 82 (1) : 93-103. Doi: 10.1007/s11240-004-6961-6
https://doi.org/10.1007/s11240-004-6961-6
 
15. Mundhara R., Rashid A. 2006. TDZ-induced triple-response and shoot formation on intact seedlings of Linum, putative role of ethylene in regeneration. Plant Sci. 170 (2) : 185-190. Doi: 10.1016/j.plantsci.2005.06.015
https://doi.org/10.1016/j.plantsci.2005.06.015
 
16. Murashige T., Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15 (3) : 473-497. Doi: 10.1111/j.1399 3054.1962.tb08052.x
https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
 
17. Obert B., Bartosova Z., Pretova A. 2005. Dihaploid production in flax by anther and ovary cultures. J. Natural Fibers. 1 (3) : 1-14. Doi: 10.1300/J395v01n03_01
https://doi.org/10.1300/J395v01n03_01
 
18. Rutkowska-Krause I., Mankowska G., Lukaszewicz M., Szopa J. 2003. Regeneration of flax (Linum usitatissimum L.) plants from anther culture and somatic tissue with increased resistance to Fusarium oxysporum. Plant Cell Rep. 22 (2) : 110-116. doi: 10.1007/s00299 003 0662-1
https://doi.org/10.1007/s00299-003-0662-1
 
19. Seta-Koselska A., Skórzyńska-Polit E. 2017. Optimization of in vitro culture conditions for obtaining flax (Linum usitatissimum L. cv. Modran) cell suspension culture. BioTechnologia. 98 (3) : 183-188. Doi: 10.5114/bta.2017.70796
https://doi.org/10.5114/bta.2017.70796
 
20. Siegień I., Adamczuk A., Wróblewska K. 2013. Light affects in vitro organogenesis of Linum usitatissimum L. and its cyanogenic potential. Acta Physiol Plant. 35 (3) : 781-789. Doi: 10.1007/s11738-012-1118-4
https://doi.org/10.1007/s11738-012-1118-4
 
21. Yildiz M., Özgen M. 2004. The effect of a submersion pretreatment on in vitro explant growth and shoot regeneration from hypocotyls of flax (Linum usitatissimum). Plant Cell Tiss. Organ Cult. 77 (1) : 111-115. Doi : 10.1023/B:TICU.0000016493.03592.c3
https://doi.org/10.1023/B:TICU.0000016493.03592.c3
 
22. Yildiz M., Sağlik C., Telci C., Erkilich E.G. 2011. The effect of in vitro competition on shoot regeneration from hypocotyl explants of Linum usitatissimum. Turk. J. Bot. 35 (2) : 211-218. Doi: 10.3906/bot-1005-26