Morphogenesis of Anthers Isolated From Cabbage (Brassica Oleracea L.) In Vitro


This study aimed to optimize the steps of obtaining regenerated cabbage plants by direct embryogenesis from isolated anthers and ovaries. Stepwise pretreatment of inflorescences was usedfor the studied hybrids and inbred lines. First, the inflorescences were placed in water and kept at a temperature of +4-6∘C for one day without the use of biologically active substances. Then the inflorescences were placed in a solution of the drug Dropp (10 mg/l) and cultivated for two days. After that, the anthers and ovaries were isolated from the flower buds and cultured on the MS culture medium at a temperature of + 32∘C for one day. The cultivation of the isolated explants on a nutrient medium (containing 0.01 mg/lof Dropp, 1.0 mg/lof NAA, 500 mg/lof asparagine, 100 mg/l of tyrosine, and 10 g/l of sucrose)led to an increase in their morphogenetic potential in the culture of anthers and ovaries (by 3.42% and 5.54%, respectively).A cytological method was usedto demonstrate the haploid nature of the regenerating plants. The number of chromosomes in the root meristem andleaves, and the chloroplasts in the closing cells of the stomatawere calculated.

Keywords: cabbage, culture in vitro, regenerated plants, anthers, ovaries, reproductive organs

[1] Petrushko YN.White cabbage. Modern resource-saving growing technology in the Primorsky Territory.Ussuriysk:AcademicPublishers; 2003.

[2] Bondareva LL, PivovarovVF.The main directions and results of selection and seed production of cabbage crops in VNIISSOK. Journalof Vegetables of Russia. 2013;3:4- 9.

[3] MayDC. Improving the technology for haploid and digaploid rape plants (Brassica napus l.) andcabbage (Brassica oleracea l.) in vitro[PhD dissertation]. Moscow, Russian Timiryazev State Agrarian University; 2010.

[4] Gil-Humanes J, Barro F. Production of doubled haploids in Brassica. Advances in haploid production in higher plants. Luxembourg, Springer, Biomedical and Life Sciences; 2009.

[5] Akmal M, Nafis T, Mirza KJ et al. High frequency somatic embryogenesis in mustard crop (Brassica juncea L. cv. Pusa Jai kisan): Microscopic and histological analyses. Australian Journal of Crop Science. 2011;5(13):1783- 1789.

[6] Na H, Hwang G, Kwak JH, Yoon MK, Chun C. Microspore derived embryo formation and doubled haploid plant production in broccoli (Brassica oleracea L. varitalica) according to nutritional and environmental conditions. African Journal of Biotechnology.2011;10(59):12535-12541.

[7] Pink D, Bailey L, McClement S et al. Double haploids, markers and QTL analysis in vegetable Brassicas. Euphytica.2008;164:509-514.

[8] Tuncer B, Ruhsar Y. Induction of microspore embryogenesis in ornamental kale by gamma irradiation and high temperature stress. Asian Journal of Biotechnology.2011;3:415-421.

[9] JongIP, Nasar UA, Hye-Ran K. (2012). Advances in in vitro culture of the Brassicaceae crop plants. Plant Biotechnology Journal. 2012;39:13–22.

[10] KalashnikovaEA, Cherednichenko MY, Kirakosyan RN, Zaitseva SM. (2017). Laboratory workshop on the culture of cells and plant tissues. Moscow: FGNU ”Rosinformagrotex”; 2017.

[11] Puchalski VA, Soloviev AA, Badaeva ED, Yrtsev VN Workshop on cytology and cytogenetics of plants. Moscow: Colossus; 2007.

[12] Muromtsev GS, Butenko RG, Tikhonenko TI, Prokofiev MI.The basics agricultural biotechnology. Moscow: Agropromizdat; 1990.

[13] Kirakosyan RN, Kalashnikova EA. (2016). Cytological and biochemical analysis of plants-regenerators of white cabbage (Brassica oleracea L.) obtained from in vitro reproductive organs. Presented at: VII International Scientific and Practical Conference dedicated to the 30th anniversary of the Department of Plant Biotechnology Nikitsky Botanical Garden; September 25th– October 1st, 2016, Yalta (Russia).

[14] Kirov I, Divashuk M, Van LaereK et al. An easy “SteamDrop” method for high quality plant chromosome preparation. Molecular Cytogenetics. 2014;7:21-21.

[15] Jahangir M, Kim HK,Choi YH, Verpoorte R. Health-affecting compounds in Brassicaceae. Comprehensive Reviews in Food Science and Food Safety. 2009;8:31–43.