A presentation prepared by Bramwel Waswa Wanjala for the ASARECA/ILRI Workshop on Mitigating the Impact of Napier Grass Smut and Stunt Diseases, Addis Ababa, June 2-3, 2010.

1. Molecular characterization of Napier grass ( Pennisetum purpureum Schumach.) germplasm using fluorescent labelled amplified fragment length polymorphism (FL-AFLP) Bramwel Waswa Wanjala End of project Workshop ILRI-ADDIS June 2010 Presented at the ASARECA/ILRI Workshop on Mitigating the Impact of Napier Grass Smut and Stunt Diseases, Addis Ababa, June 2-3, 2010

5. Pennisetum purpureum Schumach. Photo credit: Jean Hanson

7. Napier grass stunt and smut Grass stunt ( foreground ) and apparently healthy grass ( rear ) Napier infected by head smut disease Photo credit: J Hanson & M Mulaa

11. AFLP procedure Vos et al. (1995) DNA extraction Digestion of Genomic DNA and Ligation of Oligonucleotide Adapters Preselective PCR Amplification Selective PCR Amplification Capillary Electrophoresis

13. Primer-pair combination selection

14. AFLP fingerprints (Peaks) AFLP fingerprints (Peaks) for selected Primers-A,B, labelled with blue and yellow dye on sample Nap_49 and Nap_50 .Red=Polymorphic and Green=Monomorphic. Insert=Gel image

15. Genetic diversity evaluation Mean diversity estimates (H) and Shannon’s information index (I) For full data contact the authors Provenance N H I No. of polymorphic loci % Polymorphic Loci Kenya Bungoma 27 0.1169 0.2076 167 77.3 Busia 25 0.1197 0.2095 154 71.3 Butere 22 0.1492 0.2569 187 86.6 Extra 11 0.1195 0.2071 140 64.8 Mumias 35 0.1416 0.2464 193 89.4 Tanzania Hai 5 0.1747 0.2675 116 53.7 Lushoto 4 0.1727 0.2633 111 51.3 Meru 3 0.1608 0.2390 92 42.59 Muheza 14 0.1011 0.1771 128 59.26 Tarime 19 0.1893 0.3113 186 83.1 Uganda Kabarole 12 0.2130 0.3353 168 77.8 Masaka 23 0.1244 0.2099 151 69.9 Naro 5 0.0167 0.0266 13 32.5 Soroti 16 0.2142 0.3445 189 87.5 ILRI-FG Others 6 * 0.0783 0.1293 71 32.87 Swaziland 6 0.1576 0.2495 123 56.9 Tanzania 5 0.1308 0.2035 93 43.1 Unknown 14 0.1338 0.2258 145 67.1 USA 16 0.1890 0.3044 170 78.70 Zimbabwe-C 8 0.1844 0.2938 152 70.4

18. UPGMA dendrogram Summarizes genetic relationships among individuals Kenya Tanzania Uganda ILRI-FG

19. UPGMA dendrogram- Ug pop

20. Principle coordinate analysis Summarizes genetic relationships among individuals

21. AMOVA Total genetic variation is distributed between populations Source of variation df ss Ms Est.var percentage P-value All population Among population Within population 20 260 1125.941 6249.767 56.297 24.038 2.464 24.038 9% 91% 0.010 0.010 Kenya Among population Within population 4 115 105.084 2611.357 26.271 22.707 0.153 22.707 3% 99% 0.010 0.010 Uganda Among population Within population 3 52 273.893 1269.553 91.289 24.414 5.150 24.414 17% 83% 0.010 0.010 ILRI-FG Among population Within population 6 53 3.018 26.098 0.503 0.492 0.001 0.492 6% 100% 0.010 0.010

24. Acknowledgement

25. Photo credit: Eric Boa Plant Global Clinic THANK YOU