micropropagation

Megan Philpott, Valerie PenceL*, Cincinnati Zoo & Botanical Garden, United States *Speaker

Threats to global plant biodiversity compel the need for ex situ collections of species worldwide. However, the subset of species known as exceptional plants are often overlooked. These species produce few or no seeds or produce recalcitrant seeds. The Center for Conservation and Research of Endangered Wildlife (CREW) at the Cincinnati Zoo & Botanical Garden maintains a liquid nitrogen bank of exceptional plant seeds and tissues known as the CryoBioBank®. CREW has partnered with the Lyon Arboretum in Honolulu to develop cryopreservation protocols for 21 endangered exceptional Hawaiian plant species and bank them in the CryoBioBank®. To date, micropropagation protocols to produce target tissues have been tested in 13 species, and cryopreservation protocols have been tested in 7 species. As an example, in Cyrtandra gracilis, micropropagation on medium containing 2mg/L of the auxin indole-3-acetic acid and 0.1mg/L of the cytokinin 6-benzylaminopurine induces shoot organogenesis on excised leaves. These leaves were dissected into leaf bud segments and compared with shoot tips for survival after liquid nitrogen storage. Cryopreservation of shoot tips using droplet vitrification (DV) with PVS2 yielded 0% survival, while leaf bud segments yielded 20% survival. DV using PVS3 increased survival in these leaf buds to 60%. Cryopreservation of leaf bud segments immediately after bud primordia formed yielded 25% survival compared to 50% survival if the buds were left to develop into shoots before cryopreservation. Shoot tips from another species, Melicope mucronulata, have been banked in the CryoBioBank® using the DV method following an experimental survival rate of 57% after 2 days of pre- culture on a 0.3M mannitol medium instead of one day on 0.3M mannitol and one day on 0.5M mannitol medium. This project will result in the long-term protection of many endangered exceptional Hawaiian species that would otherwise be unbankable using conventional methods. (Supported by IMLS grant #MG-30-17-0055-17).

Contributing Author(s): 
Date Recorded: 
Thursday, July 25, 2019
Nellie Sugii, a key partner in the part of the collaboration focused on Hawaiian species, stands in her new micropropagation lab at Lyon Arboretum.
Photo Categories: 
Photo Credit: 
Cincinnati Zoo and Botanical Garden

Daniella DeRose, Chicago Botanic Garden

Chicago Botanic Garden’s Plants of Concern (POC) program is a collaboration between citizen scientists, natural resource managers, and researchers to collect data on rare plant populations in northeastern Illinois. The primary goal is to identify best conservation practices, while creating awareness and providing education on why conservation matters. POC engages citizen scientists, collects census data on rare plant populations—including identifying invasive species, threats, and evidence of management—and provides data to land managers. Data generated by POC inform the Illinois rare species listing process and are used by land managers to understand population trends and prioritize activities. Program monitors also serve as ecological site stewards. POC has monitored 292 species and 2453 populations, trained 953 volunteers, and worked with 135 landowners as of December 2018. POC trains its citizen scientists to collect standardized data. Partners include volunteer groups, government agencies, private landowners, and researchers.

POC is learning how management impacts rare species. Hill’s thistle, Cirsium hillii, is a habitat specialist threatened by habitat loss and lack of management. Although monitoring and management data suggested that burning and brush removal increase population size, in-depth study revealed a discrepancy between census size and effective population size. POC founder Susanne Masi led level 2 monitoring efforts looking at demography of C. hillii. CBG scientist Jeremie Fant & MS students Abigail White and Nora Gavin-Smyth found that most populations are highly clonal. Self-incompatibility, low flowering rates, and lack of compatible mates lead to low/no seed set, but genetic augmentation (introduced pollen) is able to reduce mate limitation. Based on these results, POC modified its monitoring protocol for C. hillii, introducing a minimum size for identifying individuals. CBG scientist Jacob Zeldin developed new micropropagation techniques that allow for the rapid and controlled cloning of individual C. hillii genotypes year-round with minimal space and materials. Land managers are now interested in using genetic augmentation for this species, sharing genetic material between sites.

Date Recorded: 
Thursday, May 2, 2019

Has anyone had success growing Lepanthes in micropropagation?

We are looking to propagate seeds from several Lapanthes species via micropropagation - if anyone has experience and protocols we would appreciate hearing from you.  We are working with several species, listed below, but we are looking for general information also.  

Lepanthes eltoroensis

Lepanthes rupestris

Lepanthes woodburyana

Thank you!