Abstract
To date, more than 75% of Austrian elderberry acreage is planted with cultivar ‘Haschberg’. This leads to high costs during harvest, since most of the berries have to be harvested at the same time and need to be frozen within hours after picking. In order to improve the spectrum of elderberry cultivars micropropagation of five elderberry cultivars was optimized. To meet the requirement for certified planting material, thermotherapy and chemotherapy (ribavirin), followed by meristem tip culture, were succesfully applied to eliminate various plant pathogens from elderberry plants. Virus detection was carried out by RT-PCR and confirmed by electron microscopy. An efficient protocol was developed using the cytokinins, BA for multiplication and meta-Topolin for shoot elongation and rooting of Sambucus nigra L. plants. Two commercial arbuscular mycorrhizal inoculants were evaluated for their potential to improve the process of ex-vitro-acclimatization. Both inoculants significantly enhanced survival rates, shoot length and root colonization levels of elderberry plants.
Key message
Pathogen free elderberry plants were produced via thermotherapy or chemotherapy followed by meristem culture. Culture media for micropropagation were optimized. Acclimatization of plants was significantly improved by artificial inoculation with AMF products.
This is a preview of subscription content, log in via an institution to check access.
Abbreviations
- 2iP:
-
6-(γ,γ-Dimethylallylamino)purine
- AMF:
-
Arbuscular mycorrhzial fungi
- BA:
-
N6 benzylaminopurine
- GA3 :
-
Gibberellic acid
- IBA:
-
3-Indolebutyric acid
- MSF:
-
Modified MS medium according to Finer and Nagasawa (1988)
- RT-PCR:
-
Reverse transcription polymerase chain reaction
- MS:
-
Murashige and Skoog medium (1962)
- mT:
-
N6-(3-hydroxybenzyl) adenine (meta-Topolin)
- NAA:
-
1-Napataleneacetic acid
- TDZ:
-
Thidiazuron
References
Bairu MW, Stirk WA, Dolezal K, Van Staden J (2007) Optimizing the micropropagation protocol for the endangered Aloe polyphylla: can meta-topolin and its derivatives serve as replacement for benzyladenine and zeatin? Plant Cell Tiss Organ Cult 90:15–23. https://doi.org/10.1007/s11240-007-9233-4
Bertolini E, Olmos A, López MM, Cambra M (2003) Multiplex nested reverse transcription-polymerase chain reaction in a single tube for sensitive and simultaneous detection of four RNA Viruses and Pseudomonas savastanoi pv. savastanoi in olive trees. Phytopathology 93(3):286–292. https://doi.org/10.1094/PHYTO.2003.93.3.286
Charlebois D, Brassard N (2015) Micropropagation of American elderberry: culture medium optimization and field performance. Acta Hortic 1061:175–182
Chauhan RD, Taylor NJ (2018) Meta-topolin stimulates de novo shoot organogenesis and plant regeneration in cassava. Plant Cell Tiss Organ Cult 132:219–224. https://doi.org/10.1007/s11240-017-1315-3
Ellis PJ, Converse RH, Stace-Smith R (1992) Viruses of Sambucus canadensis in North America. Acta Hortic 308:69–80. https://doi.org/10.17660/ActaHortic.1992.308.8
EPPO (2007) Diagnostic Standard PM 7/78 (1) Verticillium albo-atrum and V. dahlia on hop. Bull OEPP/EPPO Bull 37:528–535
EPPO (2008) Certification Scheme for Sambucus EPPO Bulletin 38, 19–24. https://gd.eppo.int/download/standard/111/pm4-032-1-en.pdf. Accessed 11 Sept 2018
Estrada-Luna AA, Davies FT, Egilla JN (2000) Mycorrhizal fungi enhancement of growth and gas exchange of micropropagated guava plantlets (Psidium guajava L.) during ex vitro acclimatization and plant establishment. Mykorrhiza 10:1–8. https://doi.org/10.1007/s005720050280
Faccioli G, Virus MF (1998) Virus elimination by meristem tip culture and tip micrografting. In: Hadidi A, Khetarpal RK, Koganezawa H (eds) Plant virus disease control. American Phytopathological Society Press, St Paul (MN), pp 346–380
Finer JJ, Nagasawa A (1988) Development of an embryogenic suspension culture of soybean (Glycine max. Merrill.). Plant Cell Tiss Organ Cult 15:125–138. https://doi.org/10.1007/BF00035754
Gentile A, Jàquez Gutiérrez M, Martinez J et al (2014) Effect of meta-Topolin on micropropagation and adventitious shoot regeneration in Prunus rootstocks. Plant Cell Tiss Organ Cult 118:373–381. https://doi.org/10.1007/s11240-014-0489-1
Giovannetti M, Mosse B (1980) An evaluation of techniques for measuring vesicular arbuscular mycorrhizal infection in roots. New Phytol 84:489–500. https://doi.org/10.1111/j.1469-8137.1980.tb04556.x
Harper SJ, Ward LI, Clover GRG (2010) Development of LAMP and real-time PCR methods for the rapid detection of Xylella fastidiosa for quarantine and field applications. Plant Pathol 100(12):1283–1288. https://doi.org/10.1094/PHYTO-06-10-0168
Ho T, Quito-Avila D, Keller KE, Postman JD, Martin RR, Tzanetakis IE (2016) Evidence of sympatric speciation of elderberry carlaviruses. Virus Res 215:72–75. https://doi.org/10.1016/j.virusres.2016.01.017
Hussain SA, Ahmad N, Anis M, Alatar AA (2019) Influence of meta-topolin on in vitro organogenesis in Tecoma stans L., assessment of genetic fidelity and phytochemical profiling of wild and regenerated plants. Plant Cell Tiss Organ Cult 138:339–351. https://doi.org/10.1007/s11240-019-01631-5
Jeong JJ, Ju HJ, Noh JJ (2014) A review of detection methods for the plant viruses. Res. Plant Dis. 20:173–181. https://doi.org/10.5423/RPD.2014.20.3.173
Kalinowska E, Paduch-Cichal E, Chodorska M (2013) First Report of Blueberry scorch virus in Elderberry in Poland. Plant Dis 97(11):1515. https://doi.org/10.1094/PDIS-03-13-0277-PDN
López-Fabuel I, Wetzel T, Bertolini E, Bassler A, Vidal E, Torres LB, Yuste A, Olmos A (2013) Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses. J Virol Methods 188:21–24
Monticelli S, Gentile A, Frattarelli A, Caboni E (2017) Effects of the natural cytokinin meta-Topolin on in vitro shoot proliferation and acclimatization of Prunus spp. Acta Hortic 1155:375–380. https://doi.org/10.17660/ActaHortic.2017.1155.51
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497
Newman EI (1966) A method of estimating the total length of root in a sample. J Appl Ecol 3:139–145
Panattoni A, Luvisi A, Triolo E (2013) Review. Elimination of viruses in plants: twenty years of progress. Spanish J Agric Res 11(1):173–188. https://doi.org/10.5424/sjar/2013111-3201
Rout GR, Mohapatra A, Jain SM (2006) Tissue culture of ornamental pot plant: Acritical review on present scenario and future prospects. Biotechnol Adv 24:531–560
Tang (2016a) In: Vaccinium (Blueberry & Cranberry) post-entry quarantine testing manual. New Zealand Government. https://www.mpi.govt.nz/dmsdocument/13666/. Accessed 15 May 2017.
Tang (2016b) In: Vitis (Grapevine) post-entry quarantine testing manual. https://www.mpi.govt.nz/dmsdocument/13669 Accessed 15 May 2017
Tang J, Khan S, Delmiglio C, Ward LI (2014) Sensitive detection of Tomato ringspot virus by real-time TaqMan RT-PCR targeting the highly conserved 3'-UTR region. J Virol Methods 201:38–43
Tauler M, Baraza F (2015) Improving the acclimatization and establishment of Arundo donax L. plantlets, a promising energy crop, using a mycorrhiza-based biofertilizer. Ind Crops Prod 66:299–304
Tennant D (1975) A test of a modified line intersect method of estimating root length. J Ecol 63:995–1001
Vierheilig H, Coughlan AP, Wyss U, Piché Y (1998) Ink and vinegar, a simple staining technique for arbuscular-mycorrhizal fungi. Appl Environ Microbiol 64:5004–5007
Wang GQ, Zhang T, Sun H (2012) Tissue culture and mass propagation of Sambucus willamsii Hance. J Shenyang Agric Univ. https://en.cnki.com.cn/Article_en/CJFDTOTAL-SYNY201203005.htm Accessed 14 June 2016
Werbrouck SPO, Strnad M, OnckelenH V, Debergh PC (1996) Meta-topolin, an alternative to benzyladenine in tissue culture? Physiol Plant 98:291–297. https://doi.org/10.1034/j.1399-3054.1996.980210.x
Wu QG, Zou LJ, Luo MH (2013) Callus induction and plant regeneration of Sambucus chinensis (Abstract). Zhog Yao Cai 36(12):1899–1903
Acknowledgements
We want to thank the Styrian soft fruit cooperative for supplying elderberry plants and being our partner in the Collective Research Project „Prozessoptimierung der Holunderernte durch Diversifizierung des Sortenspektrums “. This study was funded by the Austria Research Promotion Agency (FFG) (Grant number 858605). We thank Dr. Susanne Richter for electron microscopic examination of elderberry plants.
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Kopper, E., Granilshchikova, M., Leichtfried, T. et al. Micropropagation and pathogen elimination in elderberry (Sambucus nigra L.). Plant Cell Tiss Organ Cult 142, 647–652 (2020). https://doi.org/10.1007/s11240-020-01874-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-020-01874-7