Proliferation and harvesting of secondary protocorms as a novel means for improving propagation of terrestrial orchids
Betty Mauliya Bustam A B C D , Kingsley Dixon A B and Eric Bunn A B
+ Author Affiliations
- Author Affiliations
A Kings Park and Botanic Garden, Fraser Avenue, West Perth, WA 6005, Australia.
B School of Plant Biology, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia.
C Mathematics and Natural Sciences Faculty, Syiah Kuala University, Jl. Syech Abdurrauf No. 3, Darussalam, Banda Aceh 23111, Indonesia.
D Corresponding author. Email: liya1304@yahoo.com
Australian Journal of Botany 62(7) 614-621 https://doi.org/10.1071/BT14291
Submitted: 28 October 2014 Accepted: 17 January 2015 Published: 19 February 2015
Abstract
This study investigated optimisation of media and primary-protocorm development stages to enhance secondary-protocorm production as a novel means for propagation of terrestrial orchids, including taxa of conservation concern. Seeds of Caladenia latifolia were germinated asymbiotically on ½-strength Murashige and Skoog (MS) medium fortified with 5% (v/v) coconut water. Resulting protocorms at 3, 5 and 7 weeks of growth were subcultured to protocorm-proliferation media treatments consisting of ½-strength MS basal-salts medium with 6-benzylaminopurine (BA) and α- naphthaleneacetic acid (NAA) singly or in combination. Conversion of seeds to primary protocorms was high (87–92%). The highest percentage of secondary-protocorm proliferation was 40.1%, using 5-week-old protocorms (early Stage 4 of protocorm development) as explants and cultured on ½-strength MS with a combination of 5 µM NAA + 2 µM BA. Half-strength MS media containing a single plant-growth regulator (BA or NAA) were substantially less effective (<10% protocorm proliferation). The present study has provided a novel approach to sequential protocorm production that will be of value particularly for threatened orchids with limited seed availability. Protocorm proliferation in vitro enables a renewable supply of protocorms with which to conduct propagation, cryostorage and pilot restoration programs.
Additional keywords: Caladenia latifolia, conservation, protocorm proliferation, restoration.
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