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Category: BioOne.1
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Print ISSN: 1054-5476
Current: Nov 2008 : Volume 44 Issue 6
BioOne Member Since: 2004 (Active through 2008)
Frequency: Bimonthly
Impact Factor: 0.548
ISI Journal Citation Reports® Rankings:
112/152 - Plant Sciences
149/156 - Cell Biology
35/47 - Developmental Biology
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Most Read Articles (last 30 Days)
PHYSIOLOGY OF EFFECTS OF TEMPORARY IMMERSION BIOREACTORS ON MICROPROPAGATED PINEAPPLE PLANTLETS
FACTORS AFFECTING SOMATIC EMBRYOGENESIS AND PLANT REGENERATION FROM A RANGE OF RECALCITRANT GENOTYPES OF CHINESE COTTONS (GOSSYPIUM HIRSUTUM L)
Podophyllotoxin production via cell and adventitious root cultures of Podophyllum peltatum
Protocorm-like body (PLB) formation and plant regeneration from the callus culture of Dendrobium candidum Wall ex Lindl
Genetic analyses of micropropagated and regenerated plantlets of banana as assessed by RAPD and ISSR markers
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Home / All Titles / In Vitro Cellular and Developmental Biology - Plant / November 2003 / pg(s) 578-585
In Vitro Cellular and Developmental Biology - Plant
Published by: Society for In Vitro Biology
« previous article : next article »
In Vitro Cellular and Developmental Biology - Plant 39(6):578-585. 2003
doi: 10.1079/IVP2003454
TISSUE CULTURE AND AGROBACTERIUM-MEDIATED TRANSFORMATION OF HEMP (CANNABIS SATIVA L.)
M. FEENEYa and Z. K. PUNJAab
aCentre for Environmental Biology, Department of Biological Sciences, 8888 University Drive, Simon Fraser University, Burnaby, British Columbia, Canada V5A 1S6
bAuthor to whom correspondence should be addressed: punja@ sfu.ca
P. Ozias-Akins
Summary
Hemp (Cannabis sativa L.) is cultivated in many parts of the world for its fiber, oil, and seed. The development of new hemp cultivars with improved traits could be facilitated through the application of biotechnological strategies. The purpose of this study was to investigate the propagation of hemp in tissue culture and to establish a protocol for Agrobacterium-mediated transformation for foreign gene introduction. Stem and leaf segments from seedlings of four hemp varieties were placed on Murashige and Skoog medium with Gamborg B5 vitamins (MB) supplemented with 5 µM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 µM kinetin, 3% sucrose, and 8 g l−1 agar. Large masses of callus were produced within 4 wk for all cultivars. Suspension cultures were established in MB medium containing 2.5 µM 2,4-D. To promote embryogenesis or organogenesis, explants, callus, and suspension cultures derived from a range of explant sources and seedling ages were exposed to variations in the culture medium and changes to the culture environment. None of the treatments tested were successful in promoting plantlet regeneration. Suspension cells were transformed with Agrobacterium tumefaciens strain EHA101 carrying the binary vector pNOV3635 with a gene encoding phosphomannose isomerase (PMI). Transformed callus was selected on medium containing 1–2% mannose. A chlorophenol red assay was used to confirm that the PMI gene was expressed. Polymerase chain reaction and Southern hybridization detected the presence of the PMI gene. Copy number in different lines ranged from one to four.
Received: August 20, 2002; Accepted: April 30, 2003
Keywords: callus, suspension culture, Agrobacterium tumefaciens, mannose selection, phosphomannose isomerase, regeneration
Current Issue
Category: BioOne.1
Aims & Scope
Print ISSN: 1054-5476
Current: Nov 2008 : Volume 44 Issue 6
BioOne Member Since: 2004 (Active through 2008)
Frequency: Bimonthly
Impact Factor: 0.548
ISI Journal Citation Reports® Rankings:
112/152 - Plant Sciences
149/156 - Cell Biology
35/47 - Developmental Biology
Title Tools
Most Read Articles (last 30 Days)
PHYSIOLOGY OF EFFECTS OF TEMPORARY IMMERSION BIOREACTORS ON MICROPROPAGATED PINEAPPLE PLANTLETS
FACTORS AFFECTING SOMATIC EMBRYOGENESIS AND PLANT REGENERATION FROM A RANGE OF RECALCITRANT GENOTYPES OF CHINESE COTTONS (GOSSYPIUM HIRSUTUM L)
Podophyllotoxin production via cell and adventitious root cultures of Podophyllum peltatum
Protocorm-like body (PLB) formation and plant regeneration from the callus culture of Dendrobium candidum Wall ex Lindl
Genetic analyses of micropropagated and regenerated plantlets of banana as assessed by RAPD and ISSR markers
Sign up for e-alerts
RSS Feeds
Home / All Titles / In Vitro Cellular and Developmental Biology - Plant / November 2003 / pg(s) 578-585
In Vitro Cellular and Developmental Biology - Plant
Published by: Society for In Vitro Biology
« previous article : next article »
In Vitro Cellular and Developmental Biology - Plant 39(6):578-585. 2003
doi: 10.1079/IVP2003454
TISSUE CULTURE AND AGROBACTERIUM-MEDIATED TRANSFORMATION OF HEMP (CANNABIS SATIVA L.)
M. FEENEYa and Z. K. PUNJAab
aCentre for Environmental Biology, Department of Biological Sciences, 8888 University Drive, Simon Fraser University, Burnaby, British Columbia, Canada V5A 1S6
bAuthor to whom correspondence should be addressed: punja@ sfu.ca
P. Ozias-Akins
Summary
Hemp (Cannabis sativa L.) is cultivated in many parts of the world for its fiber, oil, and seed. The development of new hemp cultivars with improved traits could be facilitated through the application of biotechnological strategies. The purpose of this study was to investigate the propagation of hemp in tissue culture and to establish a protocol for Agrobacterium-mediated transformation for foreign gene introduction. Stem and leaf segments from seedlings of four hemp varieties were placed on Murashige and Skoog medium with Gamborg B5 vitamins (MB) supplemented with 5 µM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 µM kinetin, 3% sucrose, and 8 g l−1 agar. Large masses of callus were produced within 4 wk for all cultivars. Suspension cultures were established in MB medium containing 2.5 µM 2,4-D. To promote embryogenesis or organogenesis, explants, callus, and suspension cultures derived from a range of explant sources and seedling ages were exposed to variations in the culture medium and changes to the culture environment. None of the treatments tested were successful in promoting plantlet regeneration. Suspension cells were transformed with Agrobacterium tumefaciens strain EHA101 carrying the binary vector pNOV3635 with a gene encoding phosphomannose isomerase (PMI). Transformed callus was selected on medium containing 1–2% mannose. A chlorophenol red assay was used to confirm that the PMI gene was expressed. Polymerase chain reaction and Southern hybridization detected the presence of the PMI gene. Copy number in different lines ranged from one to four.
Received: August 20, 2002; Accepted: April 30, 2003
Keywords: callus, suspension culture, Agrobacterium tumefaciens, mannose selection, phosphomannose isomerase, regeneration