TY - JOUR
T1 - A reformulation of Murashige and Skoog medium (WPBS medium) improves embryogenesis, morphogenesis and transformation efficiency in temperate and tropical grasses and cereals
AU - Dalton, Sue
N1 - Funding Information:
The Institute of Biological, Environmental and Rural Sciences (IBERS) is grant aided by the UK Biotechnology and Biological Sciences Research Council (BBSRC), under the Energy Grasses & Biorefining Institute Strategic Programme (BBS/E/W/10963A01) and the EU project ‘WATBIO’ Development of improved perennial non-food biomass and bioproduct crops for water stressed environments (FP7‐311929).
Funding Information:
I would like to express my deep gratitude to Phil Morris for all his invaluable help and advice in the preparation of this manuscript. In addition I thank Kan Wang, Iowa State University for pTF102/EHA105, Candida Nibau, IBERS for pBRACT204/AGL1, Toshihiko Komari, Japan Tobacco Inc. for pTOK233/LBA4404, Seiichi Toki, National Institute of Agrobiological Sciences, Japan for pUBA, Paul Robson, IBERS for pBKS, Marcia Buanafina, Penn State University, Rakesh Bhatia, Maurice Bosch, Lorenz Fuchs, Tim Langdon, Phil Morris, Candida Nibau, Paul Robson, IBERS for the opportunity to use their genes of interest, Marionette Alana, IRRI for IR-64 and IET4786 seed, Oumaya Bouchabk?-Coussa, INRA for BD21-3 seed, the Maize Genetics Co-operation Stock Centre for A188 seed, IPK, Gatersleben for C8-A24 seed, and Ian Thomas, IBERS for Ba3081, WPBS and other cultivar seed and for creating my experiment database. I thank also Emma Timms-Taravella and Samantha Gill for their general assistance.
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/5/1
Y1 - 2020/5/1
N2 - Slow callus growth is a barrier to efficient genetic transformation in some gramineous species. A reformulation of Murashige and Skoog (MS) medium, with additional magnesium sulphate, potassium phosphate, copper sulphate, proline and glutamine, termed WPBS medium, has been developed which improves all aspects of in vitro culture when compared with MS based media. Embryogenic callus could be produced more rapidly from responsive genotypes of sixteen cereal, forage, model and energy grass species, whether using embryos, shoot tips or proliferated meristems as explants. Three species were not transformed due to contamination or unsuitable explant, but thirteen species were transformed using an identical Agrobacterium-mediated transformation, selection and regeneration protocol, including Avena sativa and Oryza sativa. Readily transformable species such as Lolium perenne, Brachypodium distachyon and Festuca arundinacea and recalcitrant species such as Lolium temulentum and Miscanthus sinensis were reliably transformed, while two new species Phalaris arundinacea and viviparous Deschampsia cespitosa were transformed at the first attempt. It is hoped that the use of WPBS media and this general transformation protocol may help to improve the efficiency of grass and cereal transformation.
AB - Slow callus growth is a barrier to efficient genetic transformation in some gramineous species. A reformulation of Murashige and Skoog (MS) medium, with additional magnesium sulphate, potassium phosphate, copper sulphate, proline and glutamine, termed WPBS medium, has been developed which improves all aspects of in vitro culture when compared with MS based media. Embryogenic callus could be produced more rapidly from responsive genotypes of sixteen cereal, forage, model and energy grass species, whether using embryos, shoot tips or proliferated meristems as explants. Three species were not transformed due to contamination or unsuitable explant, but thirteen species were transformed using an identical Agrobacterium-mediated transformation, selection and regeneration protocol, including Avena sativa and Oryza sativa. Readily transformable species such as Lolium perenne, Brachypodium distachyon and Festuca arundinacea and recalcitrant species such as Lolium temulentum and Miscanthus sinensis were reliably transformed, while two new species Phalaris arundinacea and viviparous Deschampsia cespitosa were transformed at the first attempt. It is hoped that the use of WPBS media and this general transformation protocol may help to improve the efficiency of grass and cereal transformation.
KW - embryogenic callus
KW - transformation
KW - forage grasses
KW - model grasses
KW - energy grasses
KW - cereals
KW - Transformation
KW - Cereals
KW - Model grasses
KW - Forage grasses
KW - Embryogenic callus
KW - Energy grasses
UR - http://www.scopus.com/inward/record.url?scp=85079827680&partnerID=8YFLogxK
U2 - 10.1007/s11240-020-01784-8
DO - 10.1007/s11240-020-01784-8
M3 - Article
C2 - 32308245
SN - 0167-6857
VL - 141
SP - 257
EP - 273
JO - Plant Cell, Tissue and Organ Culture
JF - Plant Cell, Tissue and Organ Culture
IS - 2
ER -