NDLI: Agrobacterium-mediated transformation using embryogenic calli in Satsuma mandarin (Citrus unshiu Marc.) cv. Miyagawa wase

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Horticulture, Environment, and Biotechnology : Volume 52, Issue 1, February 2011

Agrobacterium-mediated transformation using embryogenic calli in Satsuma mandarin (Citrus unshiu Marc.) cv. Miyagawa wase

Content Provider	Springer Nature Link
Author	Bachchu, Md. Adnan Jin, Seong Beom Park, Jeong Won Sun, Hyeon Jin Yun, Su Hyun Lee, Hyo Yeon Lee, Dong Sun Hong, Quan Chun Kim, Yong Woo Riu, Key Zung Kim, Jae Hoon
Copyright Year	2011
Abstract	Agrobacterium-mediated transformation in Satsuma mandarin (Citrus unshiu Marc.) cv. Miyagawa wase was achieved with reasonable transformation efficiency of about 22%, which was the percentage of transgenic plantlets confirmed by genomic PCR (37 plantlets/168 hygromycin-resistant calli). Embryogenic calli of Miyagawa wase were infected with Agrobacterium tumefaciens strain EHA105 harboring binary vector pCAMBIA1300 that contained miraculin gene (a taste-modifying protein) and hygromycin as a selection marker. After 5 days of co-culture in the medium containing 100 μM acetosyringone, calli were transferred to the liquid half embryogenic cell culture medium (half concentration of Murashige and Tucker’s (MT) medium modified with the addition of 500 mg·L$^{−}$ malt extract, 50 g·L$^{−}$ sucrose and 1.55 g·L$^{−}$ glutamine) with 15 mg·L$^{−}$ hygromycin and 250 mg·L$^{−}$ cefotaxime, and were cultured for two weeks. Then, the calli were grown on the solid selection medium with 20 mg·L$^{−}$ hygromycin for four weeks and 25 mg·L$^{−}$ hygromycin for another four weeks. The resistant embryos were selected and transferred to the embryo maturation medium. After 3 weeks of culture, the heart shaped embryos were transferred to the MT medium containing 1.0 mg ·L$^{−}$ GA3, 20.0 mL·L$^{−}$ coconut water, 0.02 mg·L$^{−}$ NAA and 0.0146 mg·L$^{−}$ coumarin for embryo germination. Finally the germinated embryos were cultured on MT medium containing 3.0% sucrose and 0.8% agar for growing to the normal plant. Stable integration of the transgene in the plant genome was confirmed by PCR and Southern blot analysis.
Starting Page	170
Ending Page	175
Page Count	6
File Format	PDF
ISSN	22113452
Journal	Horticulture, Environment, and Biotechnology
Volume Number	52
Issue Number	2
e-ISSN	22113460
Language	English
Publisher	Korean Society for Horticulture and Science
Publisher Date	2011-05-01
Publisher Place	Korea
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Life Sciences Plant Physiology Plant Ecology Agriculture Plant Breeding/Biotechnology
Content Type	Text
Resource Type	Article
Subject	Plant Science Horticulture Biotechnology

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Agrobacterium-mediated transformation using embryogenic calli in Satsuma mandarin (Citrus unshiu Marc.) cv. Miyagawa wase

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