Characterization of the Root-Nodulating Rhizobia Associated with Native and Non-Native Legumes in Hawai‘i and Their Potentials for Restoration of Native Wiliwili (Erythrina sandwicensis) Trees
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2020
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University of Hawaii at Manoa
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Legumes are highly nutritional, pod-bearing plants that account for more than a quarter of
all agricultural crops. In addition to being a staple of global diets, they are feasible alternatives
to nitrogenous fertilizers as they form a symbiotic relationship with rhizobia. Through biological
nitrogen fixation, rhizobia are able to transform N 2 gas into ammonia, a readily accessible form
for living organisms. Because nodulation of legumes is important for both agricultural and
native plants, it is imperative to understand which species of rhizobia associates with which
legumes. My objective in Part I of this project was to identify native and non-native legume-
rhizobia associations by sampling the nodules of 22 legume species across the island of ‘Oahu.
Following the isolation and culturing of the corresponding rhizobia within these nodules,
molecular techniques were used to isolate, amplify, and sequence the DNA. Nodules from 51
different leguminous individuals, including 22 different species, were sampled, yielding 20
different rhizobia strains from four different genera. On the island of ‘Oahu, Bradyrhizobium sp.
strain c020_F was found to be a generalist, forming nodules on nine different leguminous
species. My objective in Part II of this project was to develop an Erythrina sandwicensis
(wiliwili) restoration protocol using the two strains of Bradyrhizobium isolated from its roots in
Part I. This protocol tested the feasibility and effectiveness of sowing seeds into soil inoculated
by a rhizobia slurry of 10 6 rhizobia/mL. After three months, nodulated wiliwili seedlings had a
significantly larger biomass than non-nodulated seedlings. Both strains of Bradyrhizobium had a
nodulation rate of 80%; however, there were no significant differences in biomass between the
two strains. Results of this experiment suggest that soil inoculation of wiliwili seedlings yields
larger, healthier individuals and should be considered in future restoration protocols.
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49 pages
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