Tissue culture and plant regeneration of big cordgrass,Spartina cynosuroides: implications for wetland restoration

Abstract

Big cordgrass, Spartina cynosuroides (Poaceae), grows in extensive stands in brackish marshes along the Atlantic and Gulf coasts of the United States and along the margins of tidal streams where freshwater wetland plants dominate the back marsh. Since big cordgrass is a species of ecological importance in wetland replenishment, a tissue culture and regeneration protocol was developed t'or it. This protocol provides a mechanism for producing improved varieties. Spartina cynosuroides callus was initiated from the mesocotyl of sterile seedlings grown on both a medium containing Murashige and Skoog salts + 3% sucrose + 0.5 mg 1 -~ 6-benzylaminopurine + 1 mg I -~ 1-Naphthaleneacetic acid + 0.5 mg I ~ 2,4-diehlorophenoxyacetic acid and 5% coconut water and a medium containing Murashige and Skoog salts + 3% sucrose + 1 nag 1-~ lndole-3-acetic acid and 1 mg 1-' 2,4-dichlorophenoxyacetic acid, but not from seedlings germinated on the prcvious medium plus 5% coconut water. Shoots formed from callus after it was transferred to a shoot regeneration medium containing Murashige and Skoog salts plus 1.0 mg l '~ 6-benzytaminopurine. Roots tbrmed from the base of these shoots after the larger shoots were transferred to root regeneration medium containing reduced-strength Murashige and Skoog medium. The mode of plant regeneration was via organogenesis, and the regenerated plants have set viable seeds in the greenhouse. This callus culture and regeneration procedure will be useful for producing S. cynosuroides somaclonat variants with superior characteristics for wetland restoration and creation in stressful habitats.

Spartina alterniflora Loisel., a halophyte grass model to dissect salt stress tolerance

“…Tissue culture and plant regeneration protocols have been developed for S. alterniflora ), S. patens (Li et al 1995), and S. cynosuroides (Li and Gallagher 1996). To utilize S. alterniflora as phytoremediater, Czako et al (2006) produced transgenic lines with two genes-organomercurial lyase (mer B) and mercuric reductase (mer A)-by Agrobacterium transformation that are resistant to both organomercurials (e.g., phenyl mercuric acetate and mercuric chloride).…”

The austral cordgrass Spartina densiflora Brong.: its taxonomy, biogeography and natural history

“…Spartina Schreber (Schreber, 1789) is one of the most studied genera of salt‐marsh plants worldwide, mainly because of its abundance in space and time and because it offers a wide range of ecosystem services (Adam, 1990; Chung, 1993; Li & Gallagher, 1996). Although the cordgrass S. densiflora is presently one of the three most widely distributed species of the genus, along with S. alterniflora and S. anglica , it has not received the attention of ecologists until recently (Bortolus, 2001).…”

Transgenic Spartina alterniflora for phytoremediation

“…As none of the published protocols for in vitro regeneration of Spartina species (Croughan et al 1993;Li et al 1995;Li & Gallagher 1996;Wang et al 2003) afforded a fast growing and friable cell culture from the local, South Carolina ecotype, alternative approaches were taken. Immature embryos and seedlings of the typical Folly River area S. alterniflora were cultured on Murashige and Skoog salts with 2,4-D from 0.1-1.0 mg/L without success.…”

“…Although there were protocols published for in vitro regeneration of Spartina species (Croughan et al 1993;Li et al 1995;Li & Gallagher 1996;Wang et al 2003), local, South Carolina ecotypes (Folly River at Charleston) did not afford such cell cultures using these methods. Establishment of a cell culture and subsequent plant regeneration from the South Carolina strain was possible using alternative culture media but because of the slow growth rate the use of these cell cultures for gene transfer experiments was not practical.…”