FAITH M. MWENDWA, PROF . E. N. MAGIRI, DR. WANJOGU
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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
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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. INTRODUCTION. - PowerPoint PPT Presentation
Transcript of FAITH M. MWENDWA, PROF . E. N. MAGIRI, DR. WANJOGU
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
INTRODUCTIONRice (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.
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.
General objective
Specific objectives
1. Identify and select variant lines of Kenyan basmati 370 grown at Mwea Irrigation Scheme
2. Perform phenotypic and histochemical characterization of the lines
3. Undertake molecular characterization of the lines using SSR markers
4. Select lines with desirable qualities based on phenotypic, histochemical and molecular characterization for germplasm certification and preservation
To identify and characterize the Kenyan basmati variant lines through phenotypic and genotypic analysis
METHODOLOGY
Selection of variant lines
Planting of lines-SRI
-Paddy
-Phenotypic characterization-Histochemical test for aroma compound
Molecular characterization (SSR
primers)
Selection of lines with desirable characteristics
PHENOTYPIC CHARACTERIZATION
Qualitative data tiller typeawnsleaf colourDays 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)
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)
Control and other lines Except Line:
Plate 1: Anthocyanin colouration on stem in L2 Plate 2: No anthocyanin colouration on Stem in control
L2 CONTROL
Morphological characteristics
Plate 3: Light green leaves in L7
Plate 4: Medium green leaf colour in control
Plate 5: Dark green leaves in L12
Plate 6 : Open tillers in L13
Plate 7 : Erect tillers in control
Plate 8: Early flowering in L2 Plate 9: Flowering in control and L11
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
Plate 11: Long brown awns and tips normally
found in control
Plate 13: Long brown awns and tips normally found in control
Plate 14 and 15: Long whitish awns in L12 but brown when dry
Leaf colour, tiller type, awns and flowering dates of the lines
LINE LEAF COLOUR TILLER TYPE AWNS 1ST FLOWERING
Control Medium green Erect tillers Long, brown(2.5cm) 95 days
L1 Medium green Erect tillers Long, brown(2.0cm) 98 days
L2 Medium green Erect tillers Long, brown(2.3cm) 89 days
L3 Medium green Erect tillers Long, brown(2.1cm) 97 days
L4 Medium green Erect tillers Long ,brown(2.4cm) 96 days
L5 Medium green Erect tillers Short ,brown(1.2cm) 95 days
L6 Medium green Erect tillers Long ,brown(2.0cm) 93 days
L7 Light green Erect tillers Long ,brown(1.8cm) 96 days
L8 Medium green Erect tillers Long ,brown(1.8cm) 94 days
L9 Medium green Erect tillers Long ,brown(2.2cm) 95 days
L10 Medium green Erect tillers Long ,brown(2.0cm) 96 days
L11 Medium green Erect tillers Long ,brown(1.8cm) 95 days
L12 Dark green Erect tillers Long ,whitish(3.0cm) 96 days
L13 Medium green Open tillers No awns 97 days
L14 Medium green Erect tillers Long ,brown(1.6cm) 99 days
Control
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L1485
90
95
100
DAYS TO FLOWERING
DAYS TO FLOWERING
DAY
S
Con-trol
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L14 0
0.5
1
1.5
2
2.5
3
3.5
AWN LENGTH
AWN LENGTH
LEN
GTH
(cm
)
Quantitative parametersSRI 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)
Con-trol
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L1495
100
105
110
115
120
125
130
PLANT HEIGHT (SRI AND PADDY)
SRIPADDY
LINES
PLAN
T H
EIG
HT
(cm
)
Con-trol
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L14 10
15
20
25
30
35
40
45
50
55
60
TILLER NUMBER (SRI and PADDY)
SRIPADDY
LINES
TILL
ER N
UM
BER
Con-trol
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L14 60
65
70
75
80
85
90
95
PRODUCTIVE TILLERS (SRI and PADDY)
SRIPADDY
LINES
PRO
DU
CTIV
E T
ILLE
RS (
%)
Con-trol
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L14 15
20
25
30
35
PANICLE LENGTH (SRI and PADDY)
SRIPADDY
LINES
PAN
ICLE
LEN
GTH
(cm
)
Con-trol
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L14 30
40
50
60
70
80
90
100
110
120
130
NUMBER OF GRAINS PER PANICLE (SRI and PADDY)
SRIPADDY
LINES
NO
. O
F G
RAIN
S PE
R PA
NIC
LE
Con-trol
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L14 55
60
65
70
75
80
85
90
95
FILLED GRAINS PER PANICLE (SRI and PADDY)
SRIPADDY
LINES
FILL
ED G
RAIN
S PE
R PA
NIC
LE (
%)
Con-trol
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L14 10
15
20
25
30
35
1000 GRAIN WEIGHT (SRI and PADDY)
SRIPADDY
LINES
1000
GRA
IN W
EIG
HT
(gra
ms)
Con-trol
L1 L2 L3 L4 L5 L6 L7 L8 L9 L10 L11 L12 L13 L14 5000
55000
105000
155000
205000
255000
YIELD (SRI and PADDY)
SRIPADDY
LINES
YIEL
D (
kg/M
2)
HISTOCHEMICAL TEST FOR AROMA
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.
POSITIVE CONTROL
Orange-red regions in section
L6
L7
Plate 16,17,18: orange red colour distributed throughout the section
L2
L8
L9
Plate 19,20,21 : Colour is found near the margin
L3
L11
L12
L14
Plate 22,23,24 : Orange red spots in regions near margins
L1
Orange-red regions on margin
L4
L5
L10
Plate 25,26,27,28 : Colour found on the margins
NEGATIVE CONTROL
L13
Plate 29,30 : No reaction
MOLECULAR CHARACTERIZATION
MarkerChromosome
locationNumber of
allelesNumber f rare alleles
Size range (bp)
Highest frequency
allele PIC value
RM16 3 9 2 175-1414 175 0.837
RM161 5 9 3 177-810 211, 277 0.831
RM223 8 7 2 170-1438 530, 1438 0.812
RM72 8 3 1 101-209 209 0.550
RM171 10 5 4 374-723 374 0.42
Total 33 12
Mean 6.6 2.4 0.690
RM 16 primer
LINES Band positions due to primers (bps)RM 16 primer
1414 1347 440 365 308 289 213 194 175CONTROL + + + + + +
NON BASMATI + + + +L1 +L2 + + +L3 + + +L4 + + +L5 + + +L6 +L7 +L8 + + + + +L9 +
L10 + + + + +L11 + + + +L12 + + + + +L13 +L14 + + +
RM 161 primer
LINES Band positions due to primers (bps)RM 161 primer
810 775 705 550 523 401 277 211 177CONTROL + + + + +
NON BASMATI + + + + + +L1 + +L2 + + + + +L3 + + + + +L4 + + + + +L5 + + + +L6 + + + + +L7 + + + + +L8 + + + + +L9 + + + + + +
L10 + + + + +L11 + + + + +L12 + + + + +L13 + + +L14 + + +
LINES Band positions due to primers (bps)RM 223 primer
1438 530 413 381 187 179 170CONTROL + + + + +
NON BASMATI + + + +L1 +L2 + + + + +L3 + + + + +L4 + + +L5 + + + + +L6 + + +L7 + + +L8 + + + + +L9 + + +
L10 + + + + +L11 + + +L12 + + + +L13 +L14 +
RM 223 primer
RM 72 primer
LINES Band positions due to primers (bps)RM 72 primer
209 192 101CONTROL + +
NON BASMATI + +L1 +L2 +L3 +L4 +L5 + +L6 + +L7 + +L8 + +L9 +
L10 +L11 +L12 + +L13 + +L14 + +
RM 171 primer
LINES Band positions due to primers (bps)RM 171 primer
723 579 425 391 374CONTROL +
NON BASMATI + +L1 +L2 +L3 +L4 +L5 +L6 + +L7 +L8 +L9 +
L10 +L11 +L12 + +L13 + +L14 +
Figure : Phylogenetic tree of 14 variant rice lines based on UPGMA cluster analysis using allelic diversity data for 5 SSR markers (33 alleles)
CONCLUSIONVariant 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.
RECOMMENDATIONSFurther 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.
ACKNOWLEDGEMENTS JKUAT RPE for funding the project
Collaborators MIAD Centre Staff
Thank You
