Abstract
Papaver degenii (Urum. & Jav.) Kuzmanov is a Bulgarian glacial relict which is reported as part of Papaver alpinum subsp. alpinum by some studies. The species has restricted distribution and populations with low density and contains valuable alkaloids. Therefore, conservation measures are needed. Indirect somatic embryogenesis was induced for the first time for this species. The influence of gelling agents (Gelrite and agar) and elicitors (jasmonic acid and methyl jasmonate) on the regeneration effectiveness was studied. Phytochemical analysis on the obtained plants was conducted. It was ascertained that Murashige and Skoog medium supplemented with 4.53 μM 2,4-dichlorophenoxyacetic acid + 2.27 μM thidiazuron + 2.22 μM benzylaminopurine was the most suitable for embryogenic callus induction. The biggest number of somatic embryos during the maturation stage was formed on medium B5 solidified with Gelrite. However, medium 2B5 hardened with agar had a better effect on their further development and germination. During the germination stage, a bigger number of somatic embryos formed on the media solidified with agar. Methyl jasmonate influence depended on the stage it was applied at. It inhibited the germination of the somatic embryos when it was applied at the maturation stage and stimulated the germination when it was added at the germination stage. Jasmonic acid slightly affected the development of somatic embryos regardless of the stage of their development. Methyl jasmonate increased the percentage of alkaloid content in the developed plants. The main alkaloid in the investigated plant samples was the alkaloid amurensine.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Attree SM, Moore D, Sawhney D, Fowke LC (1991) Enhanced maturation and desiccation tolerance of white spruce (Picea glauca [Moench] Voss) somatic embryos: effects of a non-plasmolysing water stress and abscisic acid. Ann Bot 68:519–525. https://doi.org/10.1093/oxfordjournals.aob.a088291
Catana R, Holbiuc I. (2015) Direct somatic embryogenesis of the endemic taxon Papaver alpinum L. ssp. corona-sancti-stefani (Zapal.) Borza for conservative purpose. Oltenia J Stud Nat Sci 31:47–51. Available at http://olteniastudiisicomunicaristiintelenaturii.ro/cont/31_1/06_Catana.pdf
Coelho N, Gonçalves S, Romano A (2020) Endemic plant species conservation: biotechnological approaches. Plants 9:345. https://doi.org/10.3390/plants9030345
Doncheva T, Kostova N, Antonova A, Tashev A, Philipov S. (2014) Alkaloids from Papaver degenii (Urum. et Jav.) Kuzmanov. C R Acad Bulg Sci 67: 339–342. Available at http://www.proceedings.bas.bg/cgi-bin/mitko/0DOC_abs.pl?2014_3_06
Doncheva T, Stanilova M, Vutov V, Philipov S (2017) Alkaloids of seeds, in vitro cultivated and ex vitro adapted plants of the Bulgarian endemic species Papaver degenii (Papaveraceae). Nat Prod Commun 12:359–361. https://doi.org/10.1177/1934578X1701200312
Doycheva I, Philipov S, Stanilova M Influence of elicitation on Glaucium flavum cultures in in vitro conditions C R Acad Bulg Sci (in press)
Doycheva I, Yankova-Tsvetkova E, Stanilova M (2017) Somatic embryogenesis induction in Glaucium flavum Crantz (Papaveraceae) C R Acad Bulg Sci 70:525–530. Available at https://www.researchgate.net/publication/316669457_Somatic_embryogenesis_induction_in_glaucium_flavum_crantz_Papaveraceae
Eeva M, Ojala T, Tammela P, Galambosi B, Vuorela H, Hiltunen R, Fagerstedt K, Vuorela P (2003) Propagation of Angelica archangelica plants in an air-spurged bioreactor from a novel embryogenic cell line, and their production of coumarins. Biol Plant 46:343–347. https://doi.org/10.1023/A:1024309731828
Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158. https://doi.org/10.1016/0014-4827(68)90403-5
Gorgorov R, Yankova E, Baldjiev G, Apostolova I, Yurukova-Grancharova P, Stanilova M. (2011) Reproductive capacity and in vitro cultivation of the glacial relict Papaver degenii (Papaveraceae) Phyt Balc 17:333–340. Available at http://www.bio.bas.bg/~phytolbalcan/PDF/17_3/17_3_07_Gorgorov_&_al.pdf
Griffing L, Fowke LC, Constable F (1989) Radioimmunoassay of morphinan alkaloids in Papaver somniferum hypocotyls, callus, and suspension cultures. J Plant Physiol 134:645–650. https://doi.org/10.1016/S0176-1617(89)80021-5
Huang H, Liu B, Liu L, Song S (2017) Jasmonate action in plant growth and development. J Exp Bot 68:1349–1359. https://doi.org/10.1093/jxb/erw495
Ilahi I, Ghauri EG (1994) Regeneration in cultures of Papaver bracteatum hormones and temperature. Plant Cell Tiss Org Cult 38:81–83. https://doi.org/10.1007/BF00034450
Kȩpczyński J, Florek I (1997) The influence of JA-ME and ABA on induction of somatic embryogenesis in Medicago sativa L. In: Ellis RH, Black M, Murdoch AJ, Hong TD (eds) Basic and applied aspects of seed biology, Kluwer, Dordrecht, 137–140
Kim SW, In DS, Kim TJ, Liu JR (2003) High frequency somatic embryogenesis and plant regeneration in petiole and leaf explant cultures and petiole-derived embryogenic cell suspension cultures of Hylomecon vernalis. Plant Cell Tiss Org Cult 74:163–167. https://doi.org/10.1023/A:1023997627578
Kim SW, Min BW, Liu JR (1999) High frequency plant regeneration from immature ovule-derived embryogenic cell suspension cultures of Chelidonium majus var. asiaticum. Plant Cell Tiss Org Cult 56:125–129. https://doi.org/10.1023/A:1006210205704
Klimaszewska K, Bernier-Cardou M, Cyr DR, Sutton BCS (2000) Influence of gelling agents on culture medium gel strength, water availability, tissue water potential, and maturation response in embryogenic cultures of Pinus strobus L. In Vitro Cell Dev Biol - Plant 36:279–286. https://doi.org/10.1007/s11627-000-0051-1
Kuzmanov B (1970) Papaver L. In: Jordanov D (ed) Flora of the Republic of Bulgaria, 4. Publishing house of the Bulgarian Academy of Sciences, Sofia, pp 282–283
Mira MM, Wally OSD, Elhiti M, El-Shanshory A, Reddy DS, Hill RD, Stasolla C (2016) Jasmonic acid is a downstream component in the modulation of somatic embryogenesis by Arabidopsis Class 2 phytoglobin. J Exp Bot 67:2231–2246. https://doi.org/10.1093/jxb/erw022
Mohamed ME, Arafa AM, Soliman SS, Eldahmy SI (2014) Plant germination and production of callus from the yellow hornpoppy (Glaucium flavum): the first stage micropropagation. Pharmazie 69:715–720
Moon PA, Litz RE, Chávez VM (2004) Effect of gelling agent on growth and development of Ceratozamia hildae somatic embryos. Bot Rev 70:47–53. https://doi.org/10.1663/0006-8101(2004)070[0047:EOGAOG]2.0.CO;2
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Owens LD, Wozniak CA (1991) Measurement and effects of gel matric potential and expressibility on production of morphogenic callus by cultured sugarbeet leaf discs. Plant Cell Tissue Organ Cult 26:127–133. https://doi.org/10.1007/BF00036116
Palanyandy SR, Gantait S, Sinniah UR (2020) Effects of some gelling agents and their concentrations on conversion of oil palm polyembryoids into plantlets. J Genet Eng Biotechnol 18:5. https://doi.org/10.1186/s43141-019-0018-z
Park SU, Facchini PJ (2000) High-efficiency somatic embryogenesis and plant regeneration in California poppy, Eschscholzia californica Cham. Plant Cell Rep 19:421–426. https://doi.org/10.1007/s002990050750
Pathak S, Mishra BK, Misra P, Misra P, Joshi VK, Shukla S, Trivedi PK (2012) High frequency somatic embryogenesis, regeneration and correlation of alkaloid biosynthesis with gene expression in Papaver somniferum. Plant Growth Regul 68:17–25. https://doi.org/10.1007/s10725-012-9689-z
Roberts DR, Sutton BSS, Flinn BS (1990) Synchronous and high frequency germination of interior spruce somatic embryos following partial drying at high relative humidity. Can J Bot 68:1086–1090. https://doi.org/10.1139/b90-136
Rostampour S, Sohi HH, Dehestani A (2010) In vitro regeneration of Persian poppy (Papaver bracteatum). Biologia 65:647–652. https://doi.org/10.2478/s11756-010-0079-6
Rudús I, Kȩpczyńska E, Kȩpczyński J (2006) Comparative efficacy of abscisic acid and methyl jasmonate for indirect somatic embryogenesis in Medicago sativa L. Plant Growth Regul 48:1–11. https://doi.org/10.1007/s10725-005-5136-8
Ruduś I, Kępczyński J, Kępczyńska E (2001) The influence of the jasmonates and abscisic acid on callus growth and somatic embryogenesis in Medicago sativa L. tissue culture. Acta Physiol Plant 23:103–107. https://doi.org/10.1007/s11738-001-0029-6
Ruduś I, Weiler EW, Kępczyńska E (2009) Do stress-related phytohormones, abscisic acid and jasmonic acid play a role in the regulation of Medicago sativa L. somatic embryogenesis? Plant Growth Regul 59:159–169. https://doi.org/10.1007/s10725-009-9399-3
Sagare AP, Lee YL, Lin TC, Chen CC, Tsay HS (2000) Cytokinin-induced somatic embryogenesis and plant regeneration in Corydalis yanhusuo (Fumariaceae) — a medicinal plant. Plant Sci 160:139–147. https://doi.org/10.1016/S0168-9452(00)00377-0
Scholten HJ, Pierik RLM (1998) Agar as a gelling agent: chemical and physical analysis. Plant Cell Rep 17:230–235. https://doi.org/10.1007/s002990050384
Schönswetter P, Solstad H, García P, Elven R (2009) A combined molecular and morphological approach to the taxonomically intricate European mountain plant Papaver alpinum s.l. (Papaveraceae) – taxa or informal phylogeographical groups? Taxon 58:1326–1348. https://doi.org/10.1002/tax.584020
Singh A, Dwivedi P (2018) Methyl-jasmonate and salicylic acid as potent elicitors for secondary metabolite production in medicinal plants: a review. J Pharmacogn Phytochem 7:750–757. Available at https://www.phytojournal.com/archives?year=2018&vol=7&issue=1&ArticleId=2574
Singha S (1982) Influence of agar concentration on in vitro shoot proliferation of Malus spp. “Almey” and Pyrus communis “Seckel.” Am Soc Hort Sci 107:657–660
Singha S, Townsend EC, Oberly GH (1985) Mineral nutrient status of crabapple and pear shoots cultured in vitro on varying commercial agars. Am Soc Hort Sci 110:407–411
Sirmandi HB, Vahdati K, Kalantari S (2010) Enhancement of maturation and germination of somatic embryos of Persian walnut (Juglans regia L) by gellan gum, cold and PGR. Acta Hortic 861:333–344. https://doi.org/10.17660/ActaHortic.2010.861.46
Tokuji Y, Mizue Y, Masuda H (1995) Effects of methyl jasmonate and concanavalin A on embryogenesis and the induction of secondary somatic embryos of carrot. Biosi Biotech Biochem 59:1675–1678. https://doi.org/10.1271/bbb.59.1675
Tremblay L, Tremblay FM (1991) Effects of gelling agents, ammonium nitrate, and light on the development of Picea mariana (Mill) BSP. (black spruce) and Picea rubens Sarg. (red spruce) somatic embryos. Plant Sci 77:233–242. https://doi.org/10.1016/0168-9452(91)90092-M
Verrall B, Pickering CM (2020) Alpine vegetation in the context of climate change: a global review of past research and future directions. Sci Total Environ 748 https://doi.org/10.1016/j.scitotenv.2020.141344
Red Data Book of the Republic of Bulgaria, vol 1. Plants and fungi (2015) Peev D et al. (eds) BAS & MoEW, Sofia. Available at http://e-ecodb.bas.bg/rdb/en/vol1/Papdegen.html
Yang JL, Zhao B, Seong ES, Kim MJ, Kang WH, Kim NY, Yu CY, Li CH (2010) Callus induction and high-efficiency plant regeneration via somatic embryogenesis in Papaver nudicaule L. an ornamental medicinal plant. Plant Biotechnol Rep 4:261–267. https://doi.org/10.1007/s11816-010-0144-1
Zakaria RA, Hour MH, Zare N (2011) Callus production and regeneration of the medical plant Papaver orientale. Afr J Biotechnol 10:11152–11156. https://doi.org/10.5897/AJB11.204
Funding
This work was supported by the Bulgarian Ministry of Education and Science under the National Research Programme “Young scientists and postdoctoral students” approved by DCM No. 577/17.08.2018.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Doycheva, I., Doncheva, T. & Philipov, S. Indirect somatic embryogenesis induction of Papaver degenii and influence of gelling agents and elicitors. In Vitro Cell.Dev.Biol.-Plant 58, 716–727 (2022). https://doi.org/10.1007/s11627-022-10306-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11627-022-10306-4