Callus formation, plant regeneration, and transient expression in the halophyte sea aster (Aster tripolium L.).

Yuichi Uno; Shigeyuki Nakao; Yumiko Yamai; Michio Kanechi; Noboru Inagaki

Abstract  An in vitro regeneration and transient expression systems were developed for the halophyte sea aster (Aster tripolium L.), an important genetic resource for salt tolerance. Adventitious shoots were formed from both leaf explants and suspension-cultured cells in a Murashige and Skoog (MS) (Physiol Plant 15:473–497, 1962) basal salts containing 500 mg l−1 casamino acids, and supplemented with 5.4 μM a-naphthaleneacetic acid (NAA) and 4.7 μM kinetin to the culture medium. Hyperhydricity of shoots was avoided by increasing the ventilation of the culture vessel. Root formation from shoots was promoted in the presence of 26.9 μM NAA. A high yield of protoplasts was isolated using 1% cellulase and 0.25% pectinase from both leaf mesophyll and suspension-cultured cells, and these were used for transient expression. The highest level of transient expression of the green fluorescent protein was obtained with 1 × 105 protoplasts ml−1, 25 μg batch−1 of plasmid vector, and 30% polyethylene glycol 4,000.

REGENERATION (Botany); GENE expression in plants; HALOPHYTES; GERMPLASM; CELL separation; PLANT cell culture; BIOCOMPATIBILITY; MARINE plants

Plant Cell, Tissue & Organ Culture, 2009, Vol 98, Issue 3, p303

0167-6857

Academic Journal

10.1007/s11240-009-9564-4