Details
| Original language | English |
|---|---|
| Pages (from-to) | 195-205 |
| Number of pages | 11 |
| Journal | Plant cell reports |
| Volume | 32 |
| Issue number | 2 |
| Early online date | 4 Dec 2012 |
| Publication status | Published - Feb 2013 |
Abstract
Key message: The PSAG12-ipt gene was transferred to miniature rose, as the first woody species, resulting in increased ethylene resistance due to specific up-regulation of theipt gene under senescence promoting conditions. Transgenic plants of Rosa hybrida 'Linda' were obtained via transformation with Agrobacterium tumefaciens strain harboring the binary vector pSG529(+) containing the PSAG12-ipt construct. A. tumefaciens strains AGL1, GV3850 and LBA4404 (containing P35S-INTGUS gene) were used for transformation of embryogenic callus, but transgenic shoots were obtained only when AGL1 was applied. The highest transformation frequency was 10 % and it was achieved when half MS medium was used for the dilution of overnight culture of Agrobacterium. Southern blot confirmed integration of 1-6 copies of the nptII gene into the rose genome in the tested lines. Four transgenic lines were obtained which were morphologically true-to-type and indistinguishable from Wt shoots while they were in in vitro cultures. Adventitious root induction was more difficult in transgenic shoots compared to the Wt shoots, however, one of the transgenic lines (line 6) was rooted and subsequently analyzed phenotypically. The ipt expression levels were determined in this line after exposure to exogenous ethylene (3. 5 μl l-1) and/or darkness. Darkness resulted in twofold up-regulation of ipt expression, whereas darkness combined with ethylene caused eightfold up-regulation in line 6 compared to Wt plants. The transgenic line had significantly higher content of chlorophyll at the end of the treatment period compared to Wt plants.
Keywords
- Cytokinins, Ethylene, ipt, Leaf senescence, Postharvest, Rose breeding, Stress
ASJC Scopus subject areas
- Agricultural and Biological Sciences(all)
- Agronomy and Crop Science
- Agricultural and Biological Sciences(all)
- Plant Science
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In: Plant cell reports, Vol. 32, No. 2, 02.2013, p. 195-205.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Transformation of miniature potted rose (Rosa hybrida cv. Linda) with PSAG12-ipt gene delays leaf senescence and enhances resistance to exogenous ethylene
AU - Zakizadeh, Hedayat
AU - Lütken, Henrik
AU - Sriskandarajah, Sridevy
AU - Serek, Margrethe
AU - Müller, Renate
N1 - Funding Information: Acknowledgments We would like to thank Prof. R. M. Amasino, University of Wisconsin, Madison, WI, USA and Dr. Susheng Gan, Cornell University, Ithaca, New York, USA for kindly supply of the ipt construct, Mohammad Al-Salem and Dr. Heiko Mibus from University of Hannover for their help with Southern blot. This research was funded by the Ministry of Science, Research and Technology of Iran.
PY - 2013/2
Y1 - 2013/2
N2 - Key message: The PSAG12-ipt gene was transferred to miniature rose, as the first woody species, resulting in increased ethylene resistance due to specific up-regulation of theipt gene under senescence promoting conditions. Transgenic plants of Rosa hybrida 'Linda' were obtained via transformation with Agrobacterium tumefaciens strain harboring the binary vector pSG529(+) containing the PSAG12-ipt construct. A. tumefaciens strains AGL1, GV3850 and LBA4404 (containing P35S-INTGUS gene) were used for transformation of embryogenic callus, but transgenic shoots were obtained only when AGL1 was applied. The highest transformation frequency was 10 % and it was achieved when half MS medium was used for the dilution of overnight culture of Agrobacterium. Southern blot confirmed integration of 1-6 copies of the nptII gene into the rose genome in the tested lines. Four transgenic lines were obtained which were morphologically true-to-type and indistinguishable from Wt shoots while they were in in vitro cultures. Adventitious root induction was more difficult in transgenic shoots compared to the Wt shoots, however, one of the transgenic lines (line 6) was rooted and subsequently analyzed phenotypically. The ipt expression levels were determined in this line after exposure to exogenous ethylene (3. 5 μl l-1) and/or darkness. Darkness resulted in twofold up-regulation of ipt expression, whereas darkness combined with ethylene caused eightfold up-regulation in line 6 compared to Wt plants. The transgenic line had significantly higher content of chlorophyll at the end of the treatment period compared to Wt plants.
AB - Key message: The PSAG12-ipt gene was transferred to miniature rose, as the first woody species, resulting in increased ethylene resistance due to specific up-regulation of theipt gene under senescence promoting conditions. Transgenic plants of Rosa hybrida 'Linda' were obtained via transformation with Agrobacterium tumefaciens strain harboring the binary vector pSG529(+) containing the PSAG12-ipt construct. A. tumefaciens strains AGL1, GV3850 and LBA4404 (containing P35S-INTGUS gene) were used for transformation of embryogenic callus, but transgenic shoots were obtained only when AGL1 was applied. The highest transformation frequency was 10 % and it was achieved when half MS medium was used for the dilution of overnight culture of Agrobacterium. Southern blot confirmed integration of 1-6 copies of the nptII gene into the rose genome in the tested lines. Four transgenic lines were obtained which were morphologically true-to-type and indistinguishable from Wt shoots while they were in in vitro cultures. Adventitious root induction was more difficult in transgenic shoots compared to the Wt shoots, however, one of the transgenic lines (line 6) was rooted and subsequently analyzed phenotypically. The ipt expression levels were determined in this line after exposure to exogenous ethylene (3. 5 μl l-1) and/or darkness. Darkness resulted in twofold up-regulation of ipt expression, whereas darkness combined with ethylene caused eightfold up-regulation in line 6 compared to Wt plants. The transgenic line had significantly higher content of chlorophyll at the end of the treatment period compared to Wt plants.
KW - Cytokinins
KW - Ethylene
KW - ipt
KW - Leaf senescence
KW - Postharvest
KW - Rose breeding
KW - Stress
UR - http://www.scopus.com/inward/record.url?scp=84872486146&partnerID=8YFLogxK
U2 - 10.1007/s00299-012-1354-5
DO - 10.1007/s00299-012-1354-5
M3 - Article
C2 - 23207761
AN - SCOPUS:84872486146
VL - 32
SP - 195
EP - 205
JO - Plant cell reports
JF - Plant cell reports
SN - 0721-7714
IS - 2
ER -