A simple shoot multiplication procedure using internode explants,and its application for particle bombardment and Agrobacterium-mediated transformation in watercress |
| |
Authors: | Shinjiro Ogita Miki Usui Nanae Shibutani Yasuo Kato |
| |
Affiliation: | (1) Laboratory of Plant and Cell Engineering, Department of Biotechnology, Toyama Prefectural University, 5180 Kurokawa, Imizu Toyama, 939-0398, Japan |
| |
Abstract: | A shoot multiplication system derived from internode explants was investigated with the aim of improving genetic characteristics of watercress (Nasturtium officinale R. Br.). Internodes of ca. 1 cm excised from in vitro stock shoot culture were placed on half-strength Murashige and Skoog (MS) medium supplemented with 3 μM 2,4-dichlorophenoxyacetic acid as a pre-treatment. Laser scanning microscopy indicated clearly that the first sign of meristematic cell division could be seen after 1–2 days of pre-culture, and meristematic tissues multiplied along the vascular cambium of the internode segment during 7 days of culture. Multiple shoots could be obtained from more than 90% of the pre-treated explants when they were subsequently transferred to MS medium supplemented with 1 μM thidiazuron for 3 weeks. These findings indicate that pre-treatment of the internodes for 7 days promoted their capacity for organogenesis. Using this pre-treatment, frequent generation of transgenic watercress plants was achieved by adapting particle bombardment and Agrobacterium-mediated transformation techniques with a construct expressing a synthetic green florescent protein gene. |
| |
Keywords: | Agrobacterium Green florescent protein Multiple shoot Particle bombardment Watercress |
本文献已被 SpringerLink 等数据库收录! |
|