FAITH M. MWENDWA, PROF . E. N. MAGIRI, DR. WANJOGU

1. PHENOTYPIC AND MOLECULAR CHARACTERIZATION OF KENYAN BASMATI RICE VARIANT LINES: AN APPROACH TO GERMPLASM IMPROVEMENT AND PRESERVATION FAITH M. MWENDWA, PROF. E. N. MAGIRI, DR. WANJOGU

2. INTRODUCTION • Rice (Oryza sativa) cultivation is a source of livelihood for one fifth of the world’s population. • Many families around Mwea rely on rice farming as their source of livelihood. • The most preferred varieties in Kenya are basmati (pishori). • In Kenya, the quality of basmati has been declining as reflected by reduced aroma, poor grain quality and other unique characteristics.

3. The basmati rice grown in Mwea rice paddies has not been characterized. • Project was carried out to characterize the basmati variant lines currently grown in Mwea and identify those with desirable traits and preserve the germplasm. • Certified rice seeds will be generated and availed to farmers in order to increase yield hence income and alleviate poverty.

4. General objective To identify and characterize the Kenyan basmati variant lines through phenotypic and genotypic analysis Specific objectives Identify and select variant lines of Kenyan basmati 370 grown at Mwea Irrigation Scheme Perform phenotypic and histochemical characterization of the lines Undertake molecularcharacterization of the lines using SSR markers Select lines with desirable qualities based on phenotypic, histochemical and molecular characterization for germplasm certification and preservation

5. METHODOLOGY

6. PHENOTYPICCHARACTERIZATION • Qualitative data • tiller type • awns • leaf colour • Days to flowering • Quantitative data • plant height (cm) • tiller number • productive tillers (%) • panicle length (cm) • number of grains per panicle • filled grains per panicle (%) • 1000 grain weight (grams) • yield (kg/ha)

7. Control and other lines Except Line: Unique characteristics in some lines • Erect tillers • Medium green leaves • No anthocyanin colouration on stem • Awns • Brown awns • Flowered 95 ± 2 days after sowing • L13 (open tillers) • L7 (light green) and L12 (dark green) • L2 • L13 • L12 (long whitish awns) and L3 & L14 (short purple awns in some hills) but brown when grains dry • L2 (89 days after sowing) and L1 & L14 ( 98 and 99 days after sowing)

8. Morphological characteristics L2 CONTROL Plate 1: Anthocyanin colouration on stem in L2 Plate 2: No anthocyanin colouration on Stem in control

9. Plate 4: Medium green leaf colour in control Plate 3: Light green leaves in L7 Plate 5: Dark green leaves in L12

10. Plate 6 : Open tillers in L13 Plate 7 : Erect tillers in control

11. Plate 8: Early flowering in L2 Plate 9: Flowering in control and L11

12. Plate 11: Long brown awns and tips normally found in control Plate 10: Short purple awns and tips in some L3 and L14 hills Plate 12: Sample L13 had no awns and had short and broad grains

13. Plate 13: Long brown awns and tips normally found in control Plate 14 and 15: Long whitish awns in L12 but brown when dry

14. Leaf colour, tiller type, awns and flowering dates of the lines

17. SRI PADDY • Taller (105.97-124.42 cm) • More tillers (39.43-54.67 tillers) • Less productive tillers (66.41-88.53%) • Longer panicles (24.64-29.12 cm) • More grains per panicle (97.52-124.26 grains) • More filled grains per panicle (76.25-93.33%) • Heavier grains (24.65-33.48 grams for 1000 grains) • More yield (12.67-25.42 kg/ha) • Shorter (97.50-106.51 cm) • Less tillers (16.22-19.82 tillers) • More productive tillers (84.36-90.29%) • Shorter panicles (19.59-25.04 cm) • Less grains per panicle (36.88-69.19 grains) • Less filled grains per panicle(59.73-83.04%) • Lighter grains(15.25-24.84 grams for 1000 grains) • Less yield (0.87-3.53 kg/ha) Quantitative parameters

26. HISTOCHEMICAL TESTFORAROMA • 2 acetyl pyrolline is the main compound responsible for aroma in basmati rice (Bradbury et. al., 2005). 2-acetyl-1-pyrroline + 2, 4-dinitrophenylhydrazine  2-acetyl-phenyl hydrazone (Orange-red colour) • An orange red colour on sections indicated presence of 2-acetyl-pyrolline. • Thirteen of the fourteen lines had the orange red colour in the sections which varied in intensity and distribution. • L13 did not have the colour.

27. Orange-red regions in section POSITIVE CONTROL L7 L6 Plate 16,17,18: orange red colour distributed throughout the section

28. L8 L9 L2 Plate 19,20,21 : Colour is found near the margin

29. L12 L11 L3 L14 Plate 22,23,24 : Orange red spots in regions near margins

30. L4 • Orange-red regions on margin L10 L1 L5 Plate 25,26,27,28 : Colour found on the margins

31. NEGATIVE CONTROL Plate 29,30 : No reaction L13

32. MOLECULAR CHARACTERIZATION

33. RM 16 primer

34. RM 161 primer

35. RM 223 primer

36. RM 72 primer

37. RM 171 primer

38. Figure : Phylogenetic tree of 14 variant rice lines based on UPGMA cluster analysis using allelic diversity data for 5 SSR markers (33 alleles)

39. CONCLUSION • Variant lines of basmati have some characteristics like normal basmati. • L13 was high yielding but lacked the characteristics of basmati 370 rice. • L6 and L7 had much aroma, long slender half-spindle shaped grains and high yield in SRI.

40. RECOMMENDATIONS • Further characterization of the variant lines be carried out based on cooking ability, measure of amount of 2-acetyl-1-pyrroline and sequencing to establish the genetic diversity. • Further work should be on L6 and L7 focusing on molecular characterization and test of the amount of aroma compound with view of developing certified seeds.

41. ACKNOWLEDGEMENTS • JKUAT RPE for funding the project • Collaborators • MIAD Centre Staff

42. Thank You