Large-scale micropropagation of the Australian key species Gahnia radula (Cyperaceae) and its return to revegetation sites
Andrea Kodym A B F , Ian Clarke C , Cristina Aponte A , Shane Turner D E , Eric Bunn D E and John Delpratt A
+ Author Affiliations
- Author Affiliations
A Faculty of Science, The University of Melbourne, Burnley Campus, Richmond, Vic. 3121, Australia.
B Core Facility Botanical Garden & Department of Pharmacognosy, University of Vienna, Vienna, 1030, Austria.
C Royal Botanic Gardens Melbourne, Private Bag 2000, Birdwood Avenue, South Yarra, Vic. 3141, Australia.
D Kings Park and Botanic Garden, West Perth, WA 6005, Australia.
E School of Plant Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, WA 6009, Australia.
F Corresponding author. Email: andrea.kodym@univie.ac.at
Australian Journal of Botany 62(5) 417-427 https://doi.org/10.1071/BT14091
Submitted: 29 April 2014 Accepted: 26 August 2014 Published: 13 October 2014
Abstract
We report on the successful propagation of the sedge Gahnia radula (R.Br.) Benth. from seed by using plant tissue culture, and its successful establishment in the field. This keystone species, although common along parts of the eastern coast of Australia, is currently not available for revegetation because of a lack of efficient propagation methods, leading to the use of substitute species in many restoration programs. Even though seed quality is a common problem for G. radula, one population bearing filled seed was located in the near-east of Melbourne and after harvest of fruit in December 2011, seeds were successfully germinated in vitro after removal of the pericarp. Overnight soaking in sterile 10% (v/v) smoke water before culturing enhanced in vitro germination from 29.2% to 66.7%. In vitro-grown seedlings were then used as starting material for tissue-culture propagation via shoot culture. A micropropagation rate of about six new plantlets per cycle was achieved within 5–6 weeks with liquid half-strength Murashige–Skoog medium and a pulse treatment with 10 µM 6-benzylaminopurine (BAP) and 2 µM naphthalene acetic acid (NAA). Plants rooted after receiving a pulse treatment with 5 µM kinetin and were successfully acclimatised into potting mix and were ready for field planting after 5–6 months. Tube stock was planted into two field sites with minimal weed control. Survival was 98% in both cases 1 month after planting and 54% and 74% after the summer. Division of in vitro-derived plants in the nursery was very successful, with 93–96% establishment of divisions. This research highlights the important role of plant tissue culture in conserving biodiversity of native flora.
Additional keywords: dormancy, germination, in vitro, sedge.
References
Baskin JM, Baskin CC (2004) A classification system for seed dormancy. Seed Science Research 14, 1–16.| A classification system for seed dormancy.Crossref | GoogleScholarGoogle Scholar |
Brundrett M, Bougher N, Dell B, Grove T, Malajczuk N (1996) ‘Working with mycorrhizas in forestry and agriculture.’ ACIAR Monograph 32. (Australian Centre for International Agricultural Research: Canberra)
Bunn E, Senaratna T, Sivasithamparam K, Dixon KW (2005) In vitro propagation of Eucalyptus phylacis L. Johnson and K. Hill., a critically endangered relict from Western Australia. In Vitro Cellular & Developmental Biology. Plant 41, 812–815.
| In vitro propagation of Eucalyptus phylacis L. Johnson and K. Hill., a critically endangered relict from Western Australia.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XisFGqsLs%3D&md5=e0b57fbcc89497b3fd59ca00ea4a612aCAS |
Caldwell BA, Jumpponen A, Trappe JM (2000) Utilization of major detrital substrates by dark-septate, root endophytes. Mycologia 92, 230–232.
| Utilization of major detrital substrates by dark-septate, root endophytes.Crossref | GoogleScholarGoogle Scholar |
Curtis WM, Morris DI (1994) ‘Flora of Tasmania. Part 4B: Angiospermae: Alismataceae to Butmanniaceae.’ (St David’s Park Publishing: Hobart)
Electronic Flora of South Australia (2007) Fact sheet: Gahnia radula. Government of South Australia. Department of Environment, Water and Natural Resources. Available at http//:www.flora.sa.gov.au [Accessed 10 November 2013]
Elliot R, Jones D (1986) ‘Encyclopedia of Australian plants suitable for cultivation. Vol. 4.’ (Lothian Publishing: Melbourne)
Evans D, Miller M (1990) The role of the external mycelial network in the effect of soil disturbance upon vesicular – arbuscular mycorrhizal colonization of maize. New Phytologist 114, 65–71.
| The role of the external mycelial network in the effect of soil disturbance upon vesicular – arbuscular mycorrhizal colonization of maize.Crossref | GoogleScholarGoogle Scholar |
Fuchs B, Haselwandter K (2004) Red list plants: colonization by arbuscular mycorrhizal fungi and dark septate endophytes. Mycorrhiza 14, 277–281.
| Red list plants: colonization by arbuscular mycorrhizal fungi and dark septate endophytes.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD2cvhtFyiug%3D%3D&md5=97a0136201e74dd7cba716d52117d1bdCAS | 15221579PubMed |
Fuchs B, Haselwandter K (2008) Arbuscular mycorrhiza of endangered plant species: potential impacts on restoration strategies. In ‘Mycorrhiza’. (Ed. A Varma) pp. 565–579. (Springer: Berlin)
Goetghebeur P (1998) Cyperaceae. In ‘The families and genera of vascular plants. Vol. 4: flowering plants. Monocotyledons: Alismatanae and Commelinanae (except Gramineae)’. (Ed. K Kubitzki) pp. 141–190. (Springer: Berlin)
Hallaby SF, Jusaitis M (2011) Micropropagation studies of thatch saw-sedge (Gahnia radula, Cyperaceae). Report. Botanic Gardens of Adelaide, SA.
Haselwandter K, Read D (1982) The significance of a roots–fungus association in two Carex species of high-alpine plant communities. Oecologia 53, 352–354.
| The significance of a roots–fungus association in two Carex species of high-alpine plant communities.Crossref | GoogleScholarGoogle Scholar |
Hetrick BAD (1991) Mycorrhizas and root architecture. Experientia 47, 355–362.
| Mycorrhizas and root architecture.Crossref | GoogleScholarGoogle Scholar |
Jones MD, Smith SE (2004) Exploring functional definitions of mycorrhizas: are mycorrhizas always mutualisms? Canadian Journal of Botany 82, 1089–1109.
| Exploring functional definitions of mycorrhizas: are mycorrhizas always mutualisms?Crossref | GoogleScholarGoogle Scholar |
Jumpponen A (2001) Dark septate endophytes – are they mycorrhizal? Mycorrhiza 11, 207–211.
| Dark septate endophytes – are they mycorrhizal?Crossref | GoogleScholarGoogle Scholar |
Jumpponen ARI, Trappe JM (1998) Dark septate endophytes: a review of facultative biotrophic root-colonizing fungi. New Phytologist 140, 295–310.
| Dark septate endophytes: a review of facultative biotrophic root-colonizing fungi.Crossref | GoogleScholarGoogle Scholar |
Koch JM (2007) Restoring a Jarrah forest understorey vegetation after bauxite mining in Western Australia. Restoration Ecology 15, S26–S39.
| Restoring a Jarrah forest understorey vegetation after bauxite mining in Western Australia.Crossref | GoogleScholarGoogle Scholar |
Kodym A, Delpratt J (2010) Systematic investigation of fruit viability in thatch saw-sedge (Gahnia radula, Cyperaceae). Ecological Management & Restoration 11, 63–65.
| Systematic investigation of fruit viability in thatch saw-sedge (Gahnia radula, Cyperaceae).Crossref | GoogleScholarGoogle Scholar |
Kodym A, Turner S, Delpratt J (2010) In situ seed development and in vitro regeneration of three difficult-to-propagate Lepidosperma species (Cyperaceae). Australian Journal of Botany 58, 107–114.
| In situ seed development and in vitro regeneration of three difficult-to-propagate Lepidosperma species (Cyperaceae).Crossref | GoogleScholarGoogle Scholar |
Kodym A, Temsch E, Bunn E, Delpratt J (2012) Ploidy stability of somatic embryo derived plants in two ecological keystone sedge species (Lepidosperma laterale and L. concavum, Cyperaceae). Australian Journal of Botany 60, 396–404.
| Ploidy stability of somatic embryo derived plants in two ecological keystone sedge species (Lepidosperma laterale and L. concavum, Cyperaceae).Crossref | GoogleScholarGoogle Scholar |
Lagrange A, Ducousso M, Jourand P, Majorel C, Amir H (2011) New insights into the mycorrhizal status of Cyperaceae from ultramafic soils in New Caledonia. Canadian Journal of Microbiology 57, 21–28.
Lai S, Menon A, Bunn E, Turner S, Kodym A (2014) Developing an in vitro protocol for the difficult-to-propagate dryland sedge species Mesomelaena pseudostygia (Cyperaceae). In Vitro Cellular & Developmental Biology. Plant 50, 99–109.
| Developing an in vitro protocol for the difficult-to-propagate dryland sedge species Mesomelaena pseudostygia (Cyperaceae).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXjs1Cmu7k%3D&md5=8a822bf27d27f72d6de2e0a40de56737CAS |
Melbourne Water (2014) Monthly rainfall summary. Available at http://www.melbournewater.com.au/waterdata/rainfallandriverleveldata/rainfallsummary/Pages/Monthly-rainfall-summary.aspx [Accessed 2 March 2014]
Meney KA, Dixon KW (1988) Phenology, reproductive biology and seed development in four rush and sedge species from Western Australia. Australian Journal of Botany 36, 711–726.
| Phenology, reproductive biology and seed development in four rush and sedge species from Western Australia.Crossref | GoogleScholarGoogle Scholar |
Meney K, Dixon K, Scheltema M, Pate J (1993) Occurrence of vesicular mycorrhizal fungi in dryland species of Restionaceae and Cyperaceae from south-west Western Australia. Australian Journal of Botany 41, 733–737.
| Occurrence of vesicular mycorrhizal fungi in dryland species of Restionaceae and Cyperaceae from south-west Western Australia.Crossref | GoogleScholarGoogle Scholar |
Meney K, Dixon K (1995) In vitro propagation of Westerns Australian rushes (Restionaceae and realated families) by embryo culture. Part 1. In vitro embryo growth. Plant Cell, Tissue and Organ Culture 41, 107–133.
Miller RM, Smith CI, Jastrow JD, Bever JD (1999) Mycorrhizal status of the genus Carex (Cyperaceae). American Journal of Botany 86, 547–553.
| Mycorrhizal status of the genus Carex (Cyperaceae).Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3MnhtVKisw%3D%3D&md5=39030021c6be7940b4052533bb382a57CAS | 10205075PubMed |
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco callus culture. Physiologia Plantarum 15, 473–497.
| A revised medium for rapid growth and bio-assays with tobacco callus culture.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaF3sXksFKm&md5=85ecf06b1f3c63f6520567847232ed75CAS |
Muthukumar T, Kudaiyan K, Kleiner D, Manian S (1999) Mycorrhizae in sedges as related to root character and its ecological significance. Red 7, 0.31
Muthukumar T, Udaiyan K, Shanmughavel P (2004) Mycorrhiza in sedges – an overview. Mycorrhiza 14, 65–77.
| Mycorrhiza in sedges – an overview.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD2c7ovFWlug%3D%3D&md5=967de8ae0fb459569e35c9e00928ddebCAS | 14999550PubMed |
Neyland M, Bell P (2000) Ecology and conservation of the Chaostola skipper butterfly (Antipodia Chaostola leucophaea) in Tasmania. The Tasmanian Naturalist 122, 47–54.
Panaia M, Bunn E, Turner S, McComb JA (2009) Incubation temperature critical to successful stimulation of in vitro zygotic embryo growth in four Australian native Cyperaceae species. Plant Cell, Tissue and Organ Culture 97, 197–202.
| Incubation temperature critical to successful stimulation of in vitro zygotic embryo growth in four Australian native Cyperaceae species.Crossref | GoogleScholarGoogle Scholar |
Panaia M, Bunn E, McComb JA (2011) Primary and repetitive secondary somatic embryogenesis of Lepidosperma drummondii (Cyperaceae) and Baloskion tetraphyllum (Restionaceae) for land restoration and horticulture. In Vitro Cellular & Developmental Biology. Plant 47, 379–386.
| Primary and repetitive secondary somatic embryogenesis of Lepidosperma drummondii (Cyperaceae) and Baloskion tetraphyllum (Restionaceae) for land restoration and horticulture.Crossref | GoogleScholarGoogle Scholar |
Penman TD, Binns DL, Allen RM, Shiels RJ, Plummer SH (2008) Germination response of a dry sclerophyll forest soil-stored seedbank to fire related cues. Cunninghamia 10, 547–555.
Roche S, Dixon KW, Pate JS (1997) Seed ageing and smoke: partner cues in the amelioration of seed dormancy in selected Australian native species. Australian Journal of Botany 45, 783–815.
| Seed ageing and smoke: partner cues in the amelioration of seed dormancy in selected Australian native species.Crossref | GoogleScholarGoogle Scholar |
Rossetto M, Dixon KW, Meney KA, Bunn E (1992) In vitro propagation of Chinese puzzle (Caustis dioica, Cyperaceae) – a commercial sedge species from Western Australia. Plant Cell, Tissue and Organ Culture 30, 65–67.
| In vitro propagation of Chinese puzzle (Caustis dioica, Cyperaceae) – a commercial sedge species from Western Australia.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK38Xmt1ygtbY%3D&md5=a99bbca23aef8f630fa5298d106faa3aCAS |
Royal Botanic Gardens Tasmania (RTBG) (2013) ‘Achenes of Gahnia radula.’ http://www.flickr.com/photos/rtbg/12369711304/ [Accessed 21 February 2014]
Sarasan V, Cripps R, Ramsay MM, Atherton C, McMichen M, Prendercast C, Rowntree JK (2006) Conservation in vitro of threatened plants – progress in the past decade. In Vitro Cellular & Developmental Biology. Plant 42, 206–214.
| Conservation in vitro of threatened plants – progress in the past decade.Crossref | GoogleScholarGoogle Scholar |
Smith SE, Read DJ (2010) ‘Mycorrhizal symbiosis.’ (Academic Press: New York)
Terry J, Probert RJ, Linington SH (2003) Processing and maintenance of the Millennium seed bank collections. In ‘Seed conservation – turning science into practice’. (Eds RD Smith, JB Dickie, SH Linington, HW Prichard, RJ Probert) p. 318. (The Royal Botanic Gardens, Kew: London)
Turner SR (2014) Seed ecology of Lepidosperma scabrum (Cyperaceae), a dryland sedge from Western Australia with physiological seed dormancy. Australian Journal of Botany, in press.
Van Staden J, Harty AR (1988) Cyotkinins and adventitious root formation. In ‘Adventitious root formation in cuttings. Advances in Plant Sciences Series. Vol. 2’. (Eds T Davis, BE Haissig, N Sankhla) pp. 185–197. (Dioscorides Press: Portland,OR)
Watson P (2005) Saw-sedges are more than cutting grasses. Indigenotes 16, 4–5. Available at http://www.iffa.org.au/IFFA-files/Indigenotes_archive/Indigenotes_16-1_April_2005.pdf
Webber J, Johnston ME, Wearing AH (2003) High irradiance increases organogenesis in friable callus of Caustis blakei (Cyperaceae). In Vitro Cellular & Developmental Biology. Plant 39, 139–141.
| High irradiance increases organogenesis in friable callus of Caustis blakei (Cyperaceae).Crossref | GoogleScholarGoogle Scholar |
Willyams D (2005) Tissue culture of geophytic rush and sedge species for relegation of bauxite mine sites in the northern Jarrah forest of Western Australia. In ‘Contributing to a sustainable future. Proceedings of the Australian branch of the IAPTC&B, Perth, Western Australia, 21–24 September 2005’. (Eds IJ Bennet, E Bunn, H Clarke, JA McComb) pp. 226–241. (The Australasian Plant Breeding Association Inc.: Melbourne)
Wilson KL (1993) Flora of NSW: Gahnia radula. In ‘PlantNET – the plant information network system of The Royal Botanic Gardens and Domain Trust, Sydney, Australia (Version 2.0)’. Available at http://plantnet.rbgsyd.nsw.gov.au/cgibin/NSWfl.pl?page=nswfl&lvl=sp&name=Gahnia~radula [Accessed 5 March 2014]
Wilson KL (1994) Gahnia. In ‘Flora of Victoria. Vol. 2’. (Eds NG Walsh, TJ Entwisle) pp. 280–286. (Inkata Press: Melbourne)
