Abstract
An efficient method to transform five cultivars of tomato (Lycopersicon esculentum), Micro-Tom, Red Cherry, Rubion, Piedmont, and E6203 is reported. A comparison was made of leaf, cotyledon, and hypocotyl explants on 7 different regeneration media without Agrobacterium tumefaciens cocultivation and on 11 different media with cocultivation. Although all cultivars and explants formed callus and regenerated on the initial 7 media, cocultivation with A. tumefaciens significantly reduced the callus induction and regeneration. From these experiments, a transformation methodology using either hypocotyls or cotyledons cultured for one day on BA 1 mg L-1, NAA 0.1 mg L -1 and 3 days cocultivation with the Agrobacterium on this same medium followed by a transfer to a medium with zeatin 2 mg L-1 and IAA 0.1 mg L-1 for 4-6 weeks resulted in a greater than 20% transformation frequency for all five cultivars tested. In this transformation method, no feeder layers of tobacco, petunia or tomato suspension cultures were used, and the subculture media was minimal. Stable integration and transmission of the transgene in T1 generation plants were confirmed by Southern blot analysis. This procedure represents a simple, efficient and general means of transforming tomato.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1253-1257 |
| Number of pages | 5 |
| Journal | Journal of Plant Physiology |
| Volume | 160 |
| Issue number | 10 |
| DOIs | |
| State | Published - Oct 2003 |
| Externally published | Yes |
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Keywords
- Agrobacterium
- Tomato (lycopersicon esculentum)
- Transformation
- Zeatin
ASJC Scopus subject areas
- Plant Science
Cite this
Efficient and genotype-independent Agrobacterium - Mediated tomato transformation. / Park, Sung Hun; Morris, James L; Park, Jung Eun; Hirschi, Kendal D.; Smith, Roberta H.
In: Journal of Plant Physiology, Vol. 160, No. 10, 10.2003, p. 1253-1257.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Efficient and genotype-independent Agrobacterium - Mediated tomato transformation
AU - Park, Sung Hun
AU - Morris, James L
AU - Park, Jung Eun
AU - Hirschi, Kendal D.
AU - Smith, Roberta H.
PY - 2003/10
Y1 - 2003/10
N2 - An efficient method to transform five cultivars of tomato (Lycopersicon esculentum), Micro-Tom, Red Cherry, Rubion, Piedmont, and E6203 is reported. A comparison was made of leaf, cotyledon, and hypocotyl explants on 7 different regeneration media without Agrobacterium tumefaciens cocultivation and on 11 different media with cocultivation. Although all cultivars and explants formed callus and regenerated on the initial 7 media, cocultivation with A. tumefaciens significantly reduced the callus induction and regeneration. From these experiments, a transformation methodology using either hypocotyls or cotyledons cultured for one day on BA 1 mg L-1, NAA 0.1 mg L -1 and 3 days cocultivation with the Agrobacterium on this same medium followed by a transfer to a medium with zeatin 2 mg L-1 and IAA 0.1 mg L-1 for 4-6 weeks resulted in a greater than 20% transformation frequency for all five cultivars tested. In this transformation method, no feeder layers of tobacco, petunia or tomato suspension cultures were used, and the subculture media was minimal. Stable integration and transmission of the transgene in T1 generation plants were confirmed by Southern blot analysis. This procedure represents a simple, efficient and general means of transforming tomato.
AB - An efficient method to transform five cultivars of tomato (Lycopersicon esculentum), Micro-Tom, Red Cherry, Rubion, Piedmont, and E6203 is reported. A comparison was made of leaf, cotyledon, and hypocotyl explants on 7 different regeneration media without Agrobacterium tumefaciens cocultivation and on 11 different media with cocultivation. Although all cultivars and explants formed callus and regenerated on the initial 7 media, cocultivation with A. tumefaciens significantly reduced the callus induction and regeneration. From these experiments, a transformation methodology using either hypocotyls or cotyledons cultured for one day on BA 1 mg L-1, NAA 0.1 mg L -1 and 3 days cocultivation with the Agrobacterium on this same medium followed by a transfer to a medium with zeatin 2 mg L-1 and IAA 0.1 mg L-1 for 4-6 weeks resulted in a greater than 20% transformation frequency for all five cultivars tested. In this transformation method, no feeder layers of tobacco, petunia or tomato suspension cultures were used, and the subculture media was minimal. Stable integration and transmission of the transgene in T1 generation plants were confirmed by Southern blot analysis. This procedure represents a simple, efficient and general means of transforming tomato.
KW - Agrobacterium
KW - Tomato (lycopersicon esculentum)
KW - Transformation
KW - Zeatin
UR - http://www.scopus.com/inward/record.url?scp=0142229555&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0142229555&partnerID=8YFLogxK
U2 - 10.1078/0176-1617-01103
DO - 10.1078/0176-1617-01103
M3 - Article
C2 - 14610894
AN - SCOPUS:0142229555
VL - 160
SP - 1253
EP - 1257
JO - Journal of Plant Physiology
JF - Journal of Plant Physiology
SN - 0176-1617
IS - 10
ER -