Professional Documents
Culture Documents
2015
International Journal of Biosciences | IJB |
ISSN: 2220-6655 (Print), 2222-5234 (Online)
http://www.innspub.net
Vol. 6, No. 12, p. 71-80, 2015
RESEARCH PAPER
OPEN ACCESS
Gazipur-1706, Bangladesh
2
Plant Breeding Division, Bangladesh Rice Research Institute (BRRI), Gazipur, Bangladesh
http://dx.doi.org/10.12692/ijb/6.12.71-80
Abstract
The major purpose of this research was to introgress Saltol QTL into the genetic background of BRRI dhan49
through marker-assisted backcrossing. FL478 was used as a donor parent of Saltol QTL. Marker assisted
backcrossing strategies were applied to develop BRRI dhan49-Saltol lines. A primer polymorphism survey was
carried out between the two parental genotypes viz. BRRI dhan49 and FL478. A total of 363 SSR and InDel
primers were surveyed and a total of 96 markers (27%) were found polymorphic. A cross was made between
BRRI dhan49 and FL478 to produce F1 seeds and F1 was confirmed by RM493. Foreground selection was carried
out using RM493 in all generations. Fifty six SSR markers were used for background selection. The Graphical
Genotype (GGT) software was used to estimate the percentage recovery rate of the recurrent parent genome and
to find out the genome ratio of the parents in the selected progenies of the backcross populations. From BC 1F1
generation five best plants were selected based on background recovery of recurrent parent and backcrossed
with recurrent parent. From BC2F1 generation, six plants were selected based on the highest background
recovery of 80.7% to 89.5%. BC2F2 seeds were produced by selfing of selected BC2F1 individuals. The Saltol
introgressed lines performed better in salt stress condition in hydroponic screening. The developed BRRI
dhan49-Saltol lines would be useful for developing salt tolerant rice varieties or for using in other breeding
programs.
* Corresponding
71 Hoque et al.
Int. J. Biosci.
2015
Introduction
identification
al., 2002).
of
Saltol,
the
major
QTL
on
extracted
generations.
following
modified
Twelve-channel
Miniscale
pipette
was
method
used
for
This variety was derived from the cross BR4962-12-41/IR33380-7-2-1-3. The pedigree number of this
72 Hoque et al.
Int. J. Biosci.
2015
approximately.
Phenotypic evaluation
Screening of selected saltol lines were done by
Allele scoring
Data analysis
indica
origin,
and
the
lower
percentage
73 Hoque et al.
Int. J. Biosci.
2015
Fourteen
polymorphic
markers
identified
in
Final conc.
8% gel
41.35 ml
5X
6.0 ml
40% Acrylamide
8%
12 ml
10% APS
0.1%
600 l
1 l/ml
50 l
Total
60.0 ml
generation.
Chromosome 2
Chromosome 3
Chromosome 4
Chromosome 5
Chromosome 6
RM495
RM341
RM411
RM16301
RM153
RM314
RM283
RN110
RM520
RM470
RM413
RM136
RM272
RM324
RM293
RM280
RM289
RM275
RM490
RM525
RM468
RM567
RM440
RM340
RM212
RM565
RM421
RM104
RM570
RM274
RM85
RM334
Chromosome 7
Chromosome 8
Chromosome 9
Chromosome 10
Chromosome 11
Chromosome 12
RM432
RM337
RM296
RM216
RM202
RM12
RM346
RM407
RM23805
RM258
RM229
RM247
RM336
RM152
RM434
RM228
RM428
RM5556
RM23958
RM333
RM72
RM458
RM447
74 Hoque et al.
RM590
Int. J. Biosci.
2015
Table 3. Modified standard evaluation score (SES) of visual salt injury at seedling stage (IRRI, 1996).
Score
1
3
5
7
9
Observations
Normal growth, no leaf symptoms
Nearly normal growth, but leaf tips or few leaves whitish and rolled
Growth severely retarded: only a few are elongating
Complete cessation of growth; most leaves dry; some plants dying
Almost all plants are dead or dying
Classification
Highly tolerant
Tolerant
Moderately tolerant
Susceptible
Highly susceptible
generation
background
homozygous
this
with
recurrent
parent
in
BC1F1
markers
plant,
the
homozygosity
showed
used
homozygous
markers
with
showed
recurrent
with
100%
parent.
Polymorphic
Monomorphic
30
18
Nonamplification
2
3
Total
number
46
27
1
2
14
6
% Polymorphism
13
23
39
25
36
15
23
6
7
8
7
20
20
1
3
29
30
21
29
33
23
11
10
18
11
12
5
5
19
21
2
1
26
27
19.23
Total
96
246
21
363
Ave = 26.57
Highest
Rank
Lowest
Rank
30.43
2
22.22
33.33
22.22
34.78
3
2
2
27.59
23.33
24.14
24.24
38.89
1
3
1
18.52
Table 5. Performance of BRRI dhan49-Saltol lines and checks under salt stress of EC 12 dSm-1 at seedling stage
in BC2F2 generation. Data are presented as the means of three replications.
Genotypes
15-42
15-64
15-66
32-172
32-181
48-348
BRRI
dhan49
FL478
IR29
LSD(0.05)
%CV
SES
% Reduction
5.3
4.9
4.8
5.5
5.2
5.6
6.4
18.8
18.5
17.9
16.5
16.4
17.5
16.2
34.1
33.1
33.8
31.9
34.8
31.3
28.8
25.5
25.3
23.7
24.8
25.9
24.7
20.8
25.07
23.36
29.71
22.13
25.47
21.21
27.61
5.8
7.3
0.49
5.00
15.0
14.2
1.6
5.5
29.9
36.9
-
22.5
19.6
0.86
2.10
24.88
46.78
2.71
5.73
75 Hoque et al.
Root
biomass Shoot biomass (g)
(g) (Saline)
Control
Saline % Reduction
0.30
0.34
0.30
0.26
0.26
0.28
0.20
2.11
2.24
2.14
1.51
1.74
2.28
1.78
1.60
1.66
1.61
1.38
1.48
1.56
1.11
24.42
26.01
24.65
8.68
14.92
31.75
37.83
1.90
1.99
1.91
1.64
1.74
1.84
1.31
0.24
0.18
0.04
9.22
1.46
2.83
-
1.23
1.01
0.21
8.71
15.64
64.15
11.14
23.35
1.47
1.19
0.23
8.26
Int. J. Biosci.
2015
dhan49-Saltol lines.
Fig. 1. Sequential order of events for introgression of Saltol locus into the genetic background of BRRI dhan49.
Fig. 2. Foreground selection with marker RM493 for detecting Saltol QTL in BC1F1 generation. A, B, H indicate
recurrent parent, donor parent and heterozygous respectively.
Vu et al. (2012) used FL478 as donor parent of Saltol
generations.
76 Hoque et al.
Int. J. Biosci.
2015
Fig. 3. Partial view of the gel pictures of the background selection in BC1F1 generation (Note: A = Recurrent
parent: B = Donor parent; H = Heterozygous).
Fig. 4. Graphical genotyping of the plant # 3(a), plant # 7(b) in BC 1F1 generation from the cross of BRRI
dhan49*2/FL478. The red colored regions represent the homozygous regions of the recipient genome, whereas
the yellow colored regions represent the heterozygous regions.
Phenotypic evaluation of BRRI dhan49-Saltol lines
77 Hoque et al.
Int. J. Biosci.
2015
shoot
5.8 (Fig. 7). All of the test entries had reduced shoot
biomass
compared
to
FL478
in
saline
Fig. 5. Partial view of foreground selection with marker RM493 for detecting Saltol QTL in BC2F1 generation.
Sarker (2013) assessed performances of 10 BR29-
12
dSm-1
showed SES score of 4.8 and all other lines had SES
seedling stage.
78 Hoque et al.
Int. J. Biosci.
2015
Fig. 6. Graphical genotyping of the plant # 32-181, 32-172, 15-42, 15-64, 48-348 and 15-66 in BC2F1 generation
from the cross of BRRI dhan49*3/FL478. The red colored regions represent the homozygous regions of the
recipient genome, whereas the yellow colored regions represent the heterozygous regions.
79 Hoque et al.
Int. J. Biosci.
2015
Neeraja
CN,
Maghirang-Rodriguez
Pamplona A,
Heuer S,
R,
Collard BCY,
Ismail AM
assisted
backcross
approach
for
developing
symptom score.
References
Marker
Departmentof
Philippines, 399-409.
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http://dx.doi.org/10.1016/S0065-2113(07)00002-8
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http://www.plantbreeding.wur.nl/Software/ggt/ggt2
manual.pdf
Chen X, Temnykh S,
McCouch
SR.
Xu Y, Cho YG and
1997.
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of
Assisted
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(The
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80 Hoque et al.