1. + Production of Engineered long-life and male-sterile Pelargonium plants

2. + Key Words - - Pelargonium zonale and Pelargonium peltatum - - pSAG12 and PsEND1 promoter - - Engineered long-lived and male sterile plants - - ipt gene: expressed in long-living plants - - Barnase: gene expressed in pollen free plants - - Agrobacterium tumefaciens - - Delayed senescence - - Anther ablation: abnormalities in anther production

3. + Background - - Flowering plant - - Grown in Europe since 17 th century - - More than 200 species of Pelargonium - -Cultivars: P. zonale and P. peltatum is the most popular. - - Species of Pelargonium are considered important in: Market of ornamental plants Perfume industry

4. + Pelargonium Breeds Background

5. + Pelargonium breeders produce new flower colors, shapes and market quality plants via gene transfer Long-living plants Allergen-Free house plants

6. + - Agrobacterium tumefaciens - - Crown Gall Disease - - Important in gene transfer Plant Manufacturers

7. + - - In vitro regeneration system - - Pelargonium zonale explant used as a starting materials. - P. zonale – organogenesis - shoots - Two new traits in P. zonale - Long-life plants - Male sterility - Promoter used: - pSAG12 – Senescence-specific gene promoter - PsEND1 promoter – Anther specific promoter Regeneration of plants

8. + Plant senescence - - Study of aging in plants - - pSAG12 promoter fuses with ipt gene which has an ipt enzyme - - Produce more cytokinin - - Delayed Senescence

9. + Influence of pSAG12:ipt

10. + Male Sterile Plants - Why are they important? - eliminates pollen allergens - Prevents gene flow of GM material and related plant species.

11. + Elimination of pollen allergens Expression of barnase gene - Anther – specific PsEND1 promoter -Specific ablation in plants -Anther tissues are necessary - No pollen grains in ablated anthers - Barnase destroys cells for structural tissues

12. + Pelargonium zonale explant Soak it in Agrobacterium - Plasmid with T-DNA region that contains Ipt and Barnase gene under the control of their promoters Plasmid leaves bacteria, goes into plant cells DNA  chromosome = tumors Place into Media with Hormones - Roots - Shoots Transfer to a pot to fully grow

13. + RESULTS pSAG12::ipt transgenic plants Delayed leaf senscence Increased branching Smaller flowers Intense coloration Transgenic lines with PsEND1::barnase No pollen grains in anther structures

14. References BioMed Central Limited (2012, August 29). No more sneezing: Allergen-free house plants. ScienceDaily. Retrieved November 18, 2012, from http://www.sciencedaily.com­ /releases/2012/08/120831083315.htm BioMed Central Limited (31 August 2012). Production of engineered long-life and male sterile Pelargonium plants. [Research Article] Available at: http://www.biomedcentral.com/1471- 2229/12/156. [Last Accessed 18 November 2012]. Biomedcentral.com (2012) www.biomedcentral.com - Figure. [online] Available at: http://www.biomedcentral.com/1471-2229/12/156/figure/F5 [Accessed: 27 Nov 2012]. Maria Lis-Balchin, (2007). Geranium and Pelargonium: The Genera Geranium and Pelargonium. 1st ed. New York: Taylor & Francis. Mary Spink. Growing Pelargoniums. [ONLINE] Available at: : http://www.swanlandnurseries.co.uk/culturalnotes.asp. [Last Accessed 18 November 2012]. Scielo.cl (2007) Electronic Journal of Biotechnology - Influence of 2,4-D and BAP on callus growth and the subsequent regeneration of somatic embryos in long-term cultures of Pelargonium x domesticum cv. Madame Layal. [online] Available at: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000100007 [Accessed: 18 Nov 2012].