TY - JOUR

T1 - Regeneration and transformation in adult plants of Campanula species

AU - Sriskandarajah, Sridevy

AU - Mibus, Heiko

AU - Serek, Margrethe

N1 - Funding Information: Acknowledgments The project was funded by the Danish Ministry of Agriculture (93S-2466-Å97-01023), the Danish Campanula Growers: Gartneriet Thoruplund A/S (Odense) and Gartneriet El-megaard ApS (Skælskør) (SS and MS), and the University of Hannover (MS and HM). The authors thank Professor Gerco Ange-nent, Plant Research International, The Netherlands, for providing the bacterial strains and Dr. Bjarne Stummann for critical reading of the manuscript. We acknowledge Prof. Errol Hewett, Massey University, New Zealand for reading the manuscript for clarity and language. We also thank Thanussa Thuraisingam and Annette Steding for their excellent technical assistance.

PY - 2008/11

Y1 - 2008/11

N2 - Adult plants are known for recalcitrance when it comes to adventitious organ formation and regeneration. Methods used for regeneration in explants from seedlings of Campanula carpatica failed to work for explants from adult plants of the same species. The present investigation generated efficient regeneration methods for mature specimens of four species of Campanula, C. carpatica, C. haylodgensis, C. portenschlagiana and C. poscharskyana. Petiole explants from dark-grown in vitro shoot cultures grown from nodal cuttings of adult plants regenerated successfully (95%), while explants from light-grown in vitro shoot cultures and greenhouse-grown plants regenerated at 12% and zero percentage, respectively. Dark-treatment, along with media manipulation with plant growth regulators, further enhanced regenerative capacity of the explants. A MS-based medium containing 10mg l -1 TDZ and 0.25 mg l-1 NAA was the most efficient regeneration medium. Transgenic shoots from C. carpatica (3%) and C. haylodgensis (1%) and transgenic callus from all species were produced using Agrobacterium tumefaciens, and transformation was confirmed by histochemical and Southern blot analyses. Protocols developed in this study may be useful for achieving efficient regeneration and transformation of recalcitrant adult plants.

AB - Adult plants are known for recalcitrance when it comes to adventitious organ formation and regeneration. Methods used for regeneration in explants from seedlings of Campanula carpatica failed to work for explants from adult plants of the same species. The present investigation generated efficient regeneration methods for mature specimens of four species of Campanula, C. carpatica, C. haylodgensis, C. portenschlagiana and C. poscharskyana. Petiole explants from dark-grown in vitro shoot cultures grown from nodal cuttings of adult plants regenerated successfully (95%), while explants from light-grown in vitro shoot cultures and greenhouse-grown plants regenerated at 12% and zero percentage, respectively. Dark-treatment, along with media manipulation with plant growth regulators, further enhanced regenerative capacity of the explants. A MS-based medium containing 10mg l -1 TDZ and 0.25 mg l-1 NAA was the most efficient regeneration medium. Transgenic shoots from C. carpatica (3%) and C. haylodgensis (1%) and transgenic callus from all species were produced using Agrobacterium tumefaciens, and transformation was confirmed by histochemical and Southern blot analyses. Protocols developed in this study may be useful for achieving efficient regeneration and transformation of recalcitrant adult plants.

KW - Adult plants

KW - Agrobacterium tumefaciens

KW - Campanula species

KW - Regeneration

KW - Reinvigoration

KW - Transformation

UR - http://www.scopus.com/inward/record.url?scp=54049151528&partnerID=8YFLogxK

U2 - 10.1007/s00299-008-0590-1

DO - 10.1007/s00299-008-0590-1

M3 - Article

C2 - 18696077

AN - SCOPUS:54049151528

VL - 27

SP - 1713

EP - 1720

JO - Plant cell reports

JF - Plant cell reports

SN - 0721-7714

IS - 11

ER -