NDLI: Agrobacterium-mediated transformation of Pisum sativum in vitro and in vivo

Content Provider	Springer Nature Link
Author	Svabova, L. Smykal, P. Griga, M. Ondrej, V.
Copyright Year	2005
Abstract	Six pea (Pisum sativum L.) cultivars (Adept, Komet, Lantra, Olivin, Oskar, Tyrkys) were transformed via Agrobacterium tumefaciens strain EHA105 with pBIN19 plasmid carrying reporter uidA (β-glucuronidase, GUS, containing potato ST-LS1 intron) gene under the CaMV 35S promoter, and selectable marker gene nptII (neomycin phosphotransferase II) under the nos promoter. Two regeneration systems were used: continual shoot proliferation from axillary buds of cotyledonary node in vitro, and in vivo plant regeneration from imbibed germinating seed with removed testa and one cotyledon. The penetration of Agrobacterium into explants during co-cultivation was supported by sonication or vacuum infiltration treatment. The selection of putative transformants in both regeneration systems carried out on media with 100 mg dm−3 kanamycin. The presence of introduced genes was verified histochemically (GUS assay) and by means of PCR and Southern blot analysis in T0 putative transformants and their seed progenies (T1 to T3 generations). Both methods, but largely in vivo approach showed to be genotype independent, resulting in efficient and reliable transformation system for pea. The in vivo approach has in addition also benefit of time and money saving, since transgenic plants are obtained in much shorter time. All tested T0 – T3 plants were morphologically normal and fertile.
Starting Page	361
Ending Page	370
Page Count	10
File Format	PDF
ISSN	00063134
Journal	Biologia Plantarum
Volume Number	49
Issue Number	3
e-ISSN	15738264
Language	English
Publisher	Kluwer Academic Publishers-Consultants Bureau
Publisher Date	2005-01-01
Publisher Place	New York
Access Restriction	Subscribed
Subject Keyword	Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Plant Science Horticulture

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