Agravitropic behaviour of roots of rapeseed (Brassica napus L.) transformed by Agrobacterium rhizogenes.
Transgenic hairy roots of Brassica napus (cv. Omega) have been developed, using Agrobacterium rhizogenes strain AR 25, for use as a model system in the investigation of physiological and morphological differences between transgenic and normal roots. The basic parameters of growth and normal or altered gravitropical behaviour of hairy roots are for the first time presented in this paper together with an ultrastructural and morphological analysis of the root statocytes. The results obtained also represented the basis for the TRANSF0RM-experiment on the IML-2 mission performed onboard the Space Shuttle Columbia. Typical hairy root traits such as hormone-autonomous growth high growth rate, lateral branching, and changed/absence of gravitropism were detected. The transformed nature of the roots was confirmed by Southern blot analyses. The gravitropical behaviour of apices from hairy root cultures of this clone has been compared with root tips from normal seedlings. While the wild type roots curved progressively with increasing stimulation angles, the transformed roots showed no curvature when stimulated at 45 degrees, 90 degrees or 135 degrees on the ground. The morphology and ultrastructure of the root tip regions were examined by light microscopy and transmission electron microscopy. At the ultrastructural level no major differences could be detected between the roots studied. There was, however, a slight reduction in the starch content of most of the amyloplasts of the transgenic root tips, and the root cap was more V-shaped in the transgenic roots than in the wild type. Preliminary results from the Shuttle experiment TRANSFORM show a random distribution of amyloplasts in the root cells of both transformed and wild type root caps after 14 h on a 1xg centrifuge followed by 37 h in microgravity.
Odegaard
E
E
Department of Botany, Norwegian University of Science and Technology, Trondheim, Norway.
Nielsen
K M
KM
Beisvag
T
T
Evjen
K
K
Johnsson
A
A
Rasmussen
O
O
Iversen
T H
TH
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D015139:Blotting, Southern; D001937:Brassica / Q000235:genetics / Q000254:growth & development / Q000382:microbiology; D018744:DNA, Plant; D018522:Gravitropism / Q000235:genetics* / Q000502:physiology; D008854:Microscopy, Electron; D018518:Plant Root Cap / Q000254:growth & development* / Q000648:ultrastructure; D018517:Plant Roots / Q000235:genetics* / Q000254:growth & development; D030821:Plants, Genetically Modified; D010957:Plasmids; D018087:Plastids / Q000502:physiology; D013026:Space Flight; D013213:Starch; D014170:Transformation, Genetic; D014893:Weightlessness
D018744:DNA, Plant; D013213:Starch