Proliferation and harvesting of secondary protocorms as a novel means for improving propagation of terrestrial orchids
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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.
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A cryopreservation protocol for ex situ conservation of terrestrial orchids using asymbiotic primary and secondary (adventitious) protocormsBustam, B.; Dixon, Kingsley; Bunn, E. (2016)© 2015, The Society for In Vitro Biology. In a bid to better conserve endangered terrestrial orchids, we detail cryogenic research using a widely distributed terrestrial orchid, Caladenia latifolia, as a model species for ...
Cryopreservation of invitro-propagated protocorms of Caladenia for terrestrial orchid conservation in Western AustraliaWatanawikkit, P.; Tantiwiwat, S.; Bunn, E.; Dixon, Kingsley; Chayanarit, K. (2012)Cryopreservation is an important tool for the exsitu preservation of endangered plants. In this article, we describe the development of a cryopreservation protocol for orchid protocorms using the terrestrial Australian ...
Cryopreservation of secondary Protocorms, an alternative pathway for conservation of Western Australian terrestrial orchidsBustam, B.; Dixon, Kingsley; Bunn, E. (2015)Orchidaceae contains many species worldwide with a high extinction risk. Efforts to overcome this problem include ex situ approaches such as seed banking and in vitro germination of orchid seed symbiotically or asymbiotically. ...