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Title of Journal: Plant Cell Rep

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Abbravation: Plant Cell Reports

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Springer-Verlag

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DOI

10.1016/0140-6736(92)92840-C

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1432-203X

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Plant regeneration and stable transformation in th

Authors: YungTing Tsai PoYen Chen KinYing To
Publish Date: 2012/02/23
Volume: 31, Issue: 7, Pages: 1189-1198
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Abstract

Cleome spinosa is widely used as a garden ornamental in many countries Here we determined the optimal conditions for plant regeneration from different tissue explants grown in vitro Induction medium containing MS salts MS vitamins 3 sucrose 1 mg l−1 BA 200 mg l−1 timentin and 08 agar was sufficient for shoot regeneration of all the tissue explants examined including leaf hypocotyl and cotyledon Subsequently an Agrobacterium tumefaciensmediated method was developed to transform the vector pCHS which carries the transgenes Petunia chalcone synthase chs and selection marker neomycin phosphotransferase II nptII into C spinosa From a total of 368 cotyledon explants 13 putative transgenic lines were regenerated from selection medium supplemented with 50 mg l−1 kanamycin and 200 mg l−1 timentin and transferred to the greenhouse Genomic PCR and Southern blot analyses revealed that the nptII transgene was present in all 13 transgenic plants Similarly when the Petunia chs transgene was used as a probe in Southern blot analysis single or multiple hybridization bands were detected in 12 out of the 13 transgenic plants In addition T1 progeny assay from selected transformants showed that the nptII transgene can be transmitted in a Mendelian manner from transgenic parents into their progeny This is the first report of stable transformation of the C3 dicotyledon C spinosa which will facilitate functional comparison of celltype specific genes with counterpart C4 dicotyledon C gynandra using transgenic approachesThe authors thank Miranda Loney for critical reading and editing of the manuscript Director WenHsiung Li Biodiversity Research Center Academia Sinica for initiating and supporting the research project Director MingChe Shih Agricultural Biotechnology Research Center Academia Sinica Professor Maurice S B Ku Department of BioAgricultural Science National Chaiyi University Taiwan and colleagues in this research project for valuable suggestions We also thank ShowJane Sun Genomics Research Center Academia Sinica for providing and maintaining the greenhouse facility This work was financially supported by Academia Sinica Republic of China

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