Asian Journal of Agricultural Sciences 1(1): 9-11, 2009
ISSN: 2041-3890
© M axwell Scientific Organization, 2009
Mutagenic Sensitivity in Early Generation in Black Gram
1
Gaibriyal M . Lal, 1 Bini Toms and 2 Sapna Smith Lal
1
Department of Genetics and Plant Breeding
2
Division of Biochemistry, Allahabad Agricultural Institute-Deemed
University , Allah abad, Ind ia
Abstract: The investigation cond ucted with the aim of artificial ind uction of mu tation by gam ma rays to
overcome the limitation of variability in black gram and induces specific improvement without disturbing their
better attributes. The study entitled “Mutage nic sen sitivity in ea rly gen eration in black gram” was conducted
during kharif 2003 to at the field experimentation centre of the Department of Genetics and Plant Breeding.
Seeds of IPU -982 of urdb ean w ere expose d to gamm a rays and then treated w ith sodium azide to determine
their mutagenic sensitivity in terms of germination, plant survival, seedling height and pollen sterility. The
treatments with gamma rays and SA alone or in combination were more or less injurious. The combination
treatments of gamma rays and sodium azide had depressive effect on seedling grow th observed at 8 th and 12 th
days. The highe r doses/concentrations of gam ma rays and sod ium azide applied singly or in combination
severely reduced the germination, seedling height, plant survival and pollen fertility. The combination
treatments of the two mutagens produced more physiological damage as compared to the single treatments of
mutagens.
Key w ords: Mutation, gamma rays, sodium azide and black gram
INTRODUCTION
Mutation breeding is expected to make a contribution
prima rily as an important adjunct to the conventional
breeding approach. A large number of desirable varieties
have been developed through mutation bre eding in field
and horticulture crops. The urdbean, very limited work
has been carried out so far using gamma rays alone or in
combination with chemical mutagens for the improvement
of quantitative and q ualitative characters. How ever, Khan
(1984) emphasized that artificial induction of mutation by
gamma rays provides a tool to overcome the limitation of
variab ility in black gram and induces specific
improvement without disturbing their better attributes.
But the application and success of m utation breed ing in
improvement of grain legum e crops is relatively limited
except perhaps soybean and groundnut. Sodium azide has
been reported to be mutagenic in several crop species. It
is perha ps the least dangerous and the most efficient
mutagen as its yields of mutations are achieved at
mod erate M 1 sterility rates. Physiological effects of azides
are weak, few chromo somal aberrations are induced, and
it delays germ ination and growth . Gamma rays (often
denoted by Greek letter gamma) are energetic form of
electromag netic radiations produced by radioactivity or
other nuclear or sub atom ic processes with a wavelen gth
of 10 - 1 1 to 10-7 cm. They are known to be the most popular
mutage ns know n so far.
MATERIALS AND METHODS
The cultivar IPU-982 of urdbean was employed as
experimental materials during the present study. Seeds of
this variety were irradiated with 10, 20, 40 and 60 kR
doses of gam ma radiation at 6 0 Co gam ma cell. For
chem ical treatme nt, seed samples w ere presoak ed in
distilled water for 14 hours at room temperature and
treated with sodium azide at 0.01%, 0.02%, 0.03%,
0.04%, 0.05%, 0.06% and 0.07% concentrations.
Irradiated seeds along with sodium azide treated seeds
and control (untreated parental variety), were grow n in
randomized block design to study the M 1 generation
during kharif 2003-2004. Number of seeds germinated on
3 rd and 7 th day was counted and the germination percent
was calculated and the plants those survived from
germination till the maturity were counted and recorded.
The heigh t of 30 seedlings from each treatme nt was
recorded on the 8 th and 12 th day from the day of soaking
in petri dishes and compared w ith the control. Pollen
sterility was tested for each treatment by using two per
cent freshly prepared acetocarmine solution and examined
under low power microscope. Dark stained and normal
size pollen grains w ere considered as fertile and those of
irregular shape and size with light or no stain were
considered as sterile. The num ber of p lants survived till
maturity were scored from each treatment and recorded as
% survival and com pared with the con trol.
RESULTS AND DISCUSSION
Observations w ere recorded on the effect of gamma
rays and sodium azide and their different combinations on
four important characters viz., germination, plant survival,
seedling height and pollen sterility in M 1 generation.
Germination, seedling height, plant survival and pollen
sterility were gradually decreased with the increasing
Corresponding Author: Gaibriyal M Lal, Department of Genetics and Plant Breeding Agricultural Institute-Deemed
University, Allahabad
9
Asina J.Agr. Sci., 1(1): 9-11, 2009
Tab le 1:
Effect of dif ferent combinations of gamma rays and sodium azide on
plant germination and plant survival in M 1 g en er at io n of bl ac k g ra m .
Trea tmen ts
Germination
Plant s urviv al
Percentage
(%)Reduction
Percentage
(%)Reduction
0 K R+0 .0 0 % SA
90.00(27)
10.00
90.00(27)
10.00
0 K R+0 .0 1 % SA
73.33(22)
26.67
73.33(22)
26.67
0 K R+0 .0 2 % SA
60.00(18)
40.00
56.67(17)
43.33
0 K R+0 .0 3 % SA
53.33(16)
46.67
53.33(16)
46.67
0 K R+0 .0 4 % SA
50.00(15)
50.00
46.67(14)
53.33
0 K R+0 .0 5 % SA
46.67(14)
53.33
46.67(14)
53.33
0 K R+0 .0 6 % SA
43.33(13)
56.674
0.00(12)
60.00
0 K R+0 .0 7 % SA
43.33(13)
56.674
0.00(12)
60.00
1 0 K R+0 .0 0 % SA 73.33(22)
26.677
0.00(21)
30.00
1 0 K R+0 .0 1 % SA 70.00(21)
30.00
66.67(20)
33.33
1 0 K R+0 .0 2 % SA 73.33(22)
26.67
73.33(22)
26.67
1 0 K R+0 .0 3 % SA 66.67(20)
33.33
66.67(20)
33.33
1 0 K R+0 .0 4 % SA 63.33(19)
36.67
63.33(19)
36.67
1 0 K R+0 .0 5 % SA 63.33(19)
36.67
60.00(18)
40.00
1 0 K R+0 .0 6 % SA 60.00(18)
40.00
53.33(16)
46.67
1 0 K R+0 .0 7 % SA 60.00(18)
40.00
56.67(17)
43.33
2 0 K R+0 .0 0 % SA 66.67(20)
33.33
66.67(20)
33.33
2 0 K R+0 .0 1 % SA 70.00(21)
30.00
66.67(20)
33.33
2 0 K R+0 .0 2 % SA 70.00(21)
30.00
60.00(18)
40.00
2 0 K R+0 .0 3 % SA 66.67(20)
33.33
63.33(19)
36.67
2 0 K R+0 .0 4 % SA 63.33(19)
36.67
63.33(19)
36.67
2 0 K R+0 .0 5 % SA 63.33(19)
36.67
60.00(18)
40.00
2 0 K R+0 .0 6 % SA 60.00(18)
40.00
53.33(16)
46.67
2 0 K R+0 .0 7 % SA 60.00(18)
40.00
53.33(16)
46.67
4 0 K R+0 .0 0 % SA 50.00(15)
50.00
50.00(15)
50.00
4 0 K R+0 .0 1 % SA 40.00(12)
60.00
33.33(10)
66.67
4 0 K R+0 .0 2 % SA 43.33(13)
56.67
43.33(13)
56.67
4 0 K R+0 .0 3 % SA 46.67(14)
53.33
36.67(11)
63.33
4 0 K R+0 .0 4 % SA 46.67(14)
53.33
36.67(11)
63.33
4 0 K R+0 .0 5 % SA 43.33(13)
56.67
33.33(10)
66.67
4 0 K R+0 .0 6 % SA 40.00(12)
60.00
33.33(10)
66.67
4 0 K R+0 .0 7 % SA 43.33(13)
56.67
33.33(10)
66.67
6 0 K R+0 .0 0 % SA 46.67(14)
53.33
36.67(11)
63.33
6 0 K R+0 .0 1 % SA 46.67(14)
53.33
40.00(12)
60.00
6 0 K R+0 .0 2 % SA 40.00(12)
60.00
33.33(10)
66.67
6 0 K R+0 .0 3 % SA 36.67(11)
63.33
30.00(09)
70.00
6 0 K R+0 .0 4 % SA 36.67(11)
63.33
30.00(09)
70.00
6 0 K R+0 .0 5 % SA 33.33(10)
66.67
20.00(06)
80.00
6 0 K R+0 .0 6 % SA 33.33(10)
66.67
30.00(09)
70.00
6 0 K R+0 .0 7 % SA 30.00(09)
70.00
23.33(07)
76.67
Th e fig ures in pa rent hes is de note actu al ge rm inatio n an d pla nt su rviv al.
dose s/concentrations of gam ma rays and sod ium azide.
The data observed on germination percentage and
percentage reduction in M 1 generation under field
condition (Table 1) showed that there was a general
reduction in seed germination and plant survival with the
increasing doses/concentrations of different mutagens in
M 1 generation. The treatments with gamma rays and SA
alone or in combination were more or less injurious. A
reduction in germination and plant survival in the M 1
generation due to mutagenic treatments has also been
reported in pulses by Mahna et al., (1989) in Vigna
mungo. Afsar Awan et al., (1980) observed that, in
general an increase in azide con centrations resulted in
decrease in M 1 germination, the plant survival was also
affected with different doses of gamma rays and sodium
a z i d e a n d w as d ecrease d i n in c r e a sing of
dose s/concentration, which is in accordance with the
present findings. The data on seedling height recorded on
8 th and 12 th day after sowing in the laboratory (Tab le 2)
revealed that there was a marked decrease in seedling
heigh t, particularly at high doses/concentrations of
mutagens in M 1 generation as compared to the respective
controls. The m aximum reduction in the seedling height
was observed (54.64%) with 60kR+0.05%SA treatment
while the minimum reduction in seedling height (11.10%)
was found with 10k R. The combination treatments of
gamma rays and sodium azide had more depressive effect
on seedlings growth recorded on 8th and 12 th day.
The reduction in seedling height with increasing
doses of the mutagens has been established in the past by
many workers, including, Singh and Yadav (1987). The
obse rvation s on pollen sterility (Table 3) revealed that
Effect of different combinations of gamma rays and sodium azide on seedling height in M 1 generation of black gram as observed on 8 th
and 12 th day und er laboratory conditions.
8 th day
12 th day
Trea tmen ts
Seed ling heig ht (cm)
(%)Reduction
Seedling height (cm)
(%)Reduction
0KR+0.00%SA
12 .0
12 .5
0KR+0.01%SA
10 .8
10.26
11 .1
11.36
0KR+0.02%SA
9.1
24.36
9.3
25.59
0KR+0.03%SA
8.6
28.45
8.8
29.25
0KR+0.04%SA
8.5
29.14
8.8
29.35
0KR+0.05%SA
8.1
32.34
8.3
33.43
0KR+0.06%SA
7.7
35.64
7.9
36.45
0KR+0.07%SA
7.6
36.48
7.9
36.72
10KR+0.00%SA
10 .9
9.15
11 .1
11.17
10KR+0.01%SA
10 .0
16.48
10 .4
16.56
10KR+0.02%SA
8.9
25.56
9.2
26.14
10KR+0.03%SA
8.5
29.49
8.7
30.21
10KR+0.04%SA
8.1
32.46
8.4
33.17
10KR+0.05%SA
7.8
34.84
8.1
34.95
10KR+0.06%SA
7.7
35.98
7.9
36.43
10KR+0.07%SA
7.6
36.46
7.9
36.98
20KR+0.00%SA
10 .1
16.24
10 .3
17.22
20KR+0.01%SA
9.7
19.48
9.9
20.47
20KR+0.02%SA
10 .8
9.86
11 .2
10.64
20KR+0.03%SA
9.0
25.16
9.3
25.86
20KR+0.04%SA
8.7
27.16
9.1
27.46
20KR+0.05%SA
8.0
33.14
8.2
34.11
20KR+0.06%SA
7.6
36.48
7.9
37.16
20KR+0.07%SA
7.5
37.16
7.8
37.46
40KR+0.00%SA
9.7
19.46
10 .0
20.26
40KR+0.01%SA
9.5
20.46
9.8
21.41
Tab le 2:
10
Asina J.Agr. Sci., 1(1): 9-11, 2009
Table 2: Continued
40KR+0.02%SA
40KR+0.03%SA
40KR+0.04%SA
40KR+0.05%SA
40KR+0.06%SA
40KR+0.07%SA
60KR+0.00%SA
60KR+0.01%SA
60KR+0.02%SA
60KR+0.03%SA
60KR+0.04%SA
60KR+0.05%SA
60KR+0.06%SA
60KR+0.07%SA
8.8
8.4
8.1
8.0
7.7
7.4
8.9
8.5
8.0
8.0
7.6
6.2
6.9
6.4
26.64
30.15
32.16
33.49
35.65
38.04
26.18
29.46
32.98
33.48
36.48
48.45
42.16
46.48
Tab le 3: Effect of different combinations of gamma rays and sodium
azide on pollen sterility in M 1 ge ne ratio n o f bla ck gra m.
Trea tmen ts
Percentage
(%)R eduction
0KR+0.00%SA
5(1)
95
0KR+0.01%SA
10(2)
90
0KR+0.02%SA
10(2)
90
0KR+0.03%SA
20(4)
80
0KR+0.04%SA
25(5)
75
0KR+0.05%SA
30(6)
70
0KR+0.06%SA
40(8)
60
0KR+0.07%SA
40(8)
60
10KR+0.00%SA
15(3)
85
10KR+0.01%SA
15(3)
85
10KR+0.02%SA
15(3)
85
10KR+0.03%SA
20(4)
80
10KR+0.04%SA
25(5)
75
10KR+0.05%SA
35(7)
65
10KR+0.06%SA
40(8)
60
10KR+0.07%SA
45(9)
55
20KR+0.00%SA
30(6)
70
20KR+0.01%SA
30(6)
70
20KR+0.02%SA
35(7)
65
20KR+0.03%SA
40(8)
60
20KR+0.04%SA
45(9)
55
20KR+0.05%SA
45(9)
55
20KR+0.06%SA
50(10)
50
20KR+0.07%SA
55(11)
45
40KR+0.00%SA
40(8)
60
40KR+0.01%SA
45(9)
55
40KR+0.02%SA
45(9)
55
40KR+0.03%SA
55(11)
45
40KR+0.04%SA
55(11)
45
40KR+0.05%SA
55(11)
45
40KR+0.06%SA
60(12)
40
40KR+0.07%SA
60(12)
40
60KR+0.00%SA
55(11)
45
60KR+0.01%SA
60(12)
40
60KR+0.02%SA
60(12)
40
60KR+0.03%SA
60(12)
40
60KR+0.04%SA
70(14)
30
60KR+0.05%SA
75(15)
25
60KR+0.06%SA
75(15)
25
60KR+0.07%SA
75(15)
25
The figures in parenthesis denote actual pollen
9.1
7.8
8.4
8.2
8.0
7.6
8.9
8.7
8.3
8.1
7.6
5.7
6.8
5.8
27.15
30.46
32.94
34.24
36.37
38.85
28.43
30.59
33.84
35.49
39.34
54.64
45.82
53.65
in combinations or alone. The m aximum p ollen sterility
(75 .0 0% ) was observed under the treatment
60kR+0.05%SA. The combination treatment of both
gamma rays and so dium azide was found to be more
effective to produce maximum pollen sterility than when
they were applied alone. Sodium azide alone had less
effect on the pollen sterility as compared to gamma rays
alone and their combinations. The min imum pollen
sterility (10.00%) was found under 0.02%SA. In the
present findings, the increase in pollen sterility as a
consequence of mutagenesis is in accordance with the
findings of Kha n (1984), and Ignacimuthu and Babu,
(1989).
ACKNOWLEDGEMENT
The authors are highly indebted to the Hon’ble ViceChancellor Allahabad Agricultural Institute-Deemed
University, Allahabad for moral support and
enco urageme nt. Special thanks are also due to Project
Director of Nuclear Research Laboratory, IARI for
providing facilities for ga mm a radiations.
REFERENCES
Afsar, C.F. Aw an., I.N Rutger and R.A Nilan, 1980.
Mutage nic effects of Sodium Azide in Rice. Crop
Sci. 20: 661-668.
Ignacimuthu, S. and C.R. Babu, 1989. induced
chromosomal abnormality and pollen sterility in
wild and cultivated urd and mung beans.
Cytologia. 51(1): 159-167
Khan I.A., 1984. Mutation induced by gamma irradiation,
EM S and HZ in mungbean (Phasiolous aureus
Raslt) Bot. Bull. Acadenia Sinica 23(2): 105-118
Mahna S.K., R. Garg, and M. Parvateesam, 1989.
Mutage nic effects of Sodium azide in Black
gram. Currt. Sci., 58: 582-584.
Singh, V.P. and R.D.S. Yadav, 1987. Induced mutations
for quantitative and qualitative traits in green
gram and lentil. J. Genet.Plant Breed ., 45(1): 1-5
pollen sterility was much influenced by different
treatment combinations of gamma rays and sodium azide
as compared to their control. Pollen sterility was higher at
higher doses/concentrations of different mutagens either
11