International Journal of Agricultural

Science and Research (IJASR)

ISSN(P): 2250-0057; ISSN(E): 2321-0087
Vol. 6, Issue 5, Oct 2016, 105-110
TJPRC Pvt. Ltd

EFFECT OF GAMMA RAYS ON INDUCTION OF CHLOROPHYLL MUTANTS IN

GINGER GENOTYPES (ZINGIBER OFFICINALE ROSC)
P. PRASANNA KUMAR1, S. SUBRAMANIAN2, J. SURESH3, J. R. KANNAN BAPU4& R. GNANAM5
1,2,3

Department of Spices and Plantation Crops, Horticultural College and Research Institute, Coimbatore, Tamil Nadu, India
4

Department of Pulses, Centre for Plant Breeding and Genetics

Centre for Plant Molecular Biology and Biotechnology Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India

ABSTRACT
The present investigation was carried out in ginger to get the chlorophyll mutants by treating with gamma rays
conducted at the Department of Spices and Plantation Crops, Horticultural College and Research Institute, Tamil Nadu
Agricultural University, Coimbatore, during the year of 2014-2016. The experiment was laid out in Factorial Completely
Randomized Design with three replications. The gamma irradiation using Co60 source was carried out at the Department
of Plant Genetic Resource, Tamil Nadu Agricultural University. Two ginger genotypes namely Rio-de-Janeiro (G1) and
Maran (G2) were selected and their rhizome pieces were exposed to different doses of gamma rays ranging from
3,4,5,6,7,8,9,10 and 11 gray (GY) and planted in pots along with control. Different mutation frequencies and width of

ranged from 2.00 to 5.14 per cent. The spectrum of chlorophyll mutations (xantha, chlorina, albina and striata) were
observed and grouped. The overall mutation spectrum for ginger showed that xantha occurred with the highest
frequency, followed bychlorina, albina and striata.
KEYWORDS : Ginger, Gamma Rays, Chlorophyll Mutants

Original Article

mutation spectra were induced under the action of different concentrations of the gamma rays. The mutation frequency

Received: Aug 05, 2016; Accepted: Aug 29, 2016; Published: Sep 01, 2016; Paper Id.: IJASROCT201613

INTRODUCTION
Ginger (Zingiber officinale Rosc.) is an important major spice belongs to the family Zingiberaceae valued
for it aroma, flavor and also for its medicinal properties. Ginger is commercially available in various forms such
as green ginger, dry ginger, ginger powder, ginger oil, ginger oleoresin and preserved ginger
(Kizhakkayil and Sasikumar, 2009). The refreshing aroma and the pungent taste makes ginger an essential
ingredient of most world cuisine and of the food processing industry. In western countries, ginger is used in
gingerbread, biscuits, cakes, puddings, soups, pickles, beer and wine (Sanwal et al., 2010).
India is the largest producer of ginger in the world accounting for 50 per cent of the world total
production. The production trend in ginger is low in most of the countries when compared with other export crops
due to its inherent poor yields, because the ginger is not amenable to any conventional recombination breeding
programmes due to its sterility. Crop improvement of ginger is greatly challenged by poor flowering and seed set
which made improvement of this species to be confined to evolution and selection of naturally occurring variation
(Giridharan and Balakrishnan, 1992). Mutation has become a proven way of creating variation within crop
varieties and offers the possibility of inducing desired attributes which either cannot be expressed in nature or have
been lost during evolution (Iwo et al., 2012).

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P. Prasanna Kumar, S. Subramanian, J. Suresh, J. R. Kannan Bapu & R. Gnanam

The mutation breeding provides an alternative means of induction of variation. Changing the spectrum of
mutations in a predictable manner and thereby achieving directed mutagenesis is an important goal of current mutation
research (Pawar., et al 2010). Sato (1966) expressed that a close relationship exists between the chlorophyll and viable
mutations. The chlorophyllmutation frequency in mutation generation is the most dependable index for evaluating the
genetic effects of mutagenic treatments (Kharkwal, 1999; Waghmare and Mehra, 2001). Chlorophyll mutants are used as
tests for evaluation of genetic action of mutagenic factors (Svetleva, 2004). According to Brunner (1995) selection of
mutants usually begins in the second generation or later generation for the Vegetatively Propagated Plants (VPP) because it
will be much easier to recognize mutants as they segregate out.Gamma irradiation as a mutagen can induce useful as well
as harmful mutation in plants (Gupta, 1996; Micke and Donini, 1993). The aim of the work was to study the effect of
gamma rays on induction of different types of chlorophyll mutants and to determine their frequency in ginger.

MATERIALS AND METHODS

The experiment was conducted in Department of Spices and Plantation Crops, Horticultural College and Research
Institute, Tamil Nadu Agricultural University, Coimbatore during the year of 2014-2016. Two ginger genotypes
Rio-de-Janeiro and Maran were collected for this study from Kerala Agricultural University at Thrissur. Physical mutagen
(gamma ray) was employed in the present investigation. Gamma ray source was cobalt - 60 in 1000 Ci, emitting 5000 rads
per minute at the time of irradiation. The gamma irradiation using Co60 source was carried out at the Department of Plant
Genetic Resource, Tamil Nadu Agricultural University. Uniform sized rhizomes (approximately 29g) were selected and cut
into pieces. These rhizome pieceswere subjected to gamma irradiation and used as planting material for the present
investigation. These rhizome pieces were exposed to different doses of gamma rays ranging from 0, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13 and 15gray (Gy). 9 Gy was fixed as the LD50 value (the dosage which resulted in a 50 per cent survival rate).
Later the dosage were fixed from 3, 4, 5, 6, 7, 8, 9, 10 and 11 gray (Gy) and the irradiated rhizomes were planted in the pot
along with control (untreated rhizomes) for the VM1 generation. The rhizomes harvested in VM1 were used for raising VM2
generation. In both generation (VM1 and VM2) it was observed for the chlorophyll mutants. The experiment was laid out in
a Factorial Completely Randomized Design and replicated three times.
Chlorophyll mutation is observed when the seedlings were 50 days old and classified according to Gustafsson
(1940) as below.
Xantha
These mutants were distinguished by their uniform yellow colour with total absence of chlorophyll and the clones
are viable
Chlorina
These mutants had uniform yellow green colour. They emerged along the green seedlings and survive upto
maturity.
Albina
They were characterized by total absence of either chlorophyll or carotenoids and the clones are partially viable
Striata
Leaves are with green streaks and the clones are viable.
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Effect of Gamma Rays on Induction of Chlorophyll

Mutants in Ginger Genotypes (Zingiber Officinale Rosc.)

107

RESULTS AND DISCUSSIONS

In VM1 generation no chlorophyll mutant was observed in all the treatments. However in VM2 almost all the
mutagenic treatments showed different degree of mutants with respective dose. In the present study, there were differences
in the frequency of chlorophyll mutations due to the increased doses of gamma rays. The frequency of chlorophyll mutants
was high 5.14 per cent in the treatment T4 (6Gy) and 4.42 per cent inT3 (5Gy) in Rio-de-Janeiro and 4.94 per cent in T3
(5Gy) and 4.00 per cent in T4(6Gy) in Maran respectively. The occurrence of chlorophyll chimera in the present
investigation could be attributed to chromosomal aberrations, change in the route of auxin synthesis, distribution or
disruption of mineral metabolism or accumulation of free amino acids. It may also be due to the multicellular nature of the
tissues treated. Nuclear and or plastid mutations were thought to cause variation in leaves (Kirk and Bassett, 1967). Laxmi
et al. (1980) considered that chimera formation in leaves as a result of gamma irradiation might be due to the multicellular
nature of the tissues treated. Leaf variation is due to gamma irradiation had been documented in Canna
(Nakornthap, 1965). Raju et al. (1980) obtained such a phenomenon in ginger. Variation in leaf shape and colour had been
observed in costus by Gupta et al. (1982). In ginger, Giridharan (1984) recorded yellow streaks as a result of radiation
treatments in the cultivars Rio-de-Janeiro and Maran.
In the genotype G1 (Rio-de-Janeiro), the mutation frequency ranged between 2.00 and 5.14 per cent. A higher
mutation frequency (5.14 per cent) was observed in T4(6 Gy). In the genotype G2 (Maran), the mutation frequency ranged
between 2.00 and 4.94 per cent. Treatment T3 (5 Gy) recorded higher mutation frequency (4.94 per cent)
(Table 1; Figure 1).
The spectrum of chlorophyll mutants showed four different types viz., xantha, albina, chlorina and striata.
The spectrum varied with increasing dose of gamma rays. A high per cent of xantha (34.50) was recorded. This was
followed by chlorina (28.33per cent) albina (22.22 per cent) and striata (14.67 per cent), respectively.
In the genotype G1 (Rio-de-Janeiro), a higher mean percentage of xantha (34.19 per cent) was observed. This was
followed by albina(28.46 per cent), striata (18.97 per cent) and chlorina (18.97 per cent) respectively. In G2 (Maran),
greater percentage of chlorina (40.54) was obtained followed by xantha (34.86 per cent), albina (14.94 per cent) and striata
(9.66 per cent) respectively (Table 2). Pawar et al. (2010) reported that mutagenic effectiveness decreased with the
increase in dose of mutagen indicating that negative relationship between effectiveness and dose of mutagen
(Table 2; Figure 2). Thohirah et al. (2009) reported that the effect of gamma irradiation had excerted various effects on
growth of C. alismatifolia include in flower development, colour intensity, double inflorescence within one stalks as well
as chlorophyll mutation on leaves. Chlorophyll development seems to be controlled by many genes located on several
chromosomes, which could beadjacent to centromere and proximalsegment of chromosomes (Swaminathan, 1964).

CONCLUSIONS
Present investigation revealed how chlorophyll gene response to mutagen gamma rays. Chlorophyll genes are
reflected in the vM2 in the form of different types of chlorophyll mutants which can be useful as marker in physiological
and biochemical investigations.
REFERENCES
1.

Brunner, H. (1995). Methods of induction of mutations. Plant Breeding Unit. Joint FAO/ IAEA Programme, IAEA
Laboratories, Seibersdorf, Austria

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P. Prasanna Kumar, S. Subramanian, J. Suresh, J. R. Kannan Bapu & R. Gnanam

2.

Giridharan, M .P. (1984). Effect of gamma irradiation in ginger (Zingiber officinale Rosc.) M.Sc. (Hort.) thesis, Kerala
Agricultural University, Vellanikkara, India

3.

Giridharan, M.P., & Balakrishnan, S. (1992). Gamma ray induced variability in vegetative and floral characters of ginger.
Indian Cocoa, Arecanut and Spices J., 15: 68-72

4.

Kirk, J. T. O., & Tilney Bassett, R. A. E. (1967). The Plastids. W.H. Freeman and Co., San Francisco

5.

Gupta, M. N., Lakshmi, V., Dixit, V. S., & Srivastava, S. N. (1982). Gamma ray induced variability in Costus speciosus. Prog.
Hort., 14(4): 193-197

6.

Gustaffson, A. (1940). The mutation system of the chlorophyll apparatus. Lands Univ. Arskr. 36:1-40

7.

Iwo, G. A., Amadi, C. O., & Udo, I. A. (2012). Agronomic performance of ginger mutant clones derived from gamma-ray
radiation. In: Proc. 36th Annual conference of Genetics society of Nigeria held in Oct, 2012 at the University of Calabar,
Nigeria

8.

Kharkwal, M. C. (1999). Induced mutations in chickpea (Cicer arietinum L.) III. Frequency and spectrum of viable mutations.
Indian Journal of Genetics and Plant Breeding, 4: 451-464

9.

Kizhakkayil, J., & Sasikumar, B. (2009). Variability for quality traits in a global germplasm collection of ginger (Zingiber
officinale R.). Curr. Trends Biotech. Pharmacy (online), 3: 1-4

10. Laxmi, V., Gupta, M. N., Shukla, P., Dixit, B. S., & Srivastava, S. N. 1980. Effect of gamma irradiation on growth and
diosgenin content of Costus speciosus. Indian Drugs, 17(11): 371-75
11. Nakornthap, A. (1965). Radiation induced somatic mutations in the ornamental canna. Radiation Botany, 5: 707-712
12. Pawar, N., Sandeep, P., Mansingraj, N., Firdose, K., & Ghansham, D. (2010). Journals of Agriculture, 22(5): 406-411
13. Raju, E. C., Patel, J. D., & Shah, J. J. (1980). Effects of gamma irradiation in morphology of leaf and shoot apex of ginger,
turmeric and mango ginger. Proc. Indian Acad. Sci., 89(3): 173-178
14. Sato, M. (1966). Induction of mutation in rice by some chemical mutagens. Mutation induced by radiation and chemicals.
Gamma field symp.No.5
15. Sanwal, S. K., Rai, N., Singh, J., & Buragohain, J. (2010). Antioxidant phytochemicals and gingerol content in diploid and
tetraploid clones of ginger (Zingiber officinale Roscoe). Sci. Hort. 4: 280-285
16. Svetleva, D. L. (2004). Induction of chlorophyll mutants in common bean under the action of chemical mutagens ENU and
EMS. J. Central Euro. Agri., 5(2): 85-90
17. Swaminathan, M. S. (1964). A comparison of mutation induction in diploids and polyploids. In: the use of unduced mutations
in plant breeding. Rad. Mut. Organ. FAO/IAEA, Vienna. pp. 619-641
18. Thohirah, L.A., Johari, E., Mohd, E., & Nazir, B. (2009). Changes in flower Development,
19. Chlorophyll mutation and Alteration in plant morphology of Curcuma alismatifolia by gamma-irradiation. American Journal
of Applied Science, 6(7):1436 -1439
20. Waghmare, V. N., & Mehra, R. B. (2001). Induced chlorophyll mutations, mutagenic effectiveness and efficiency in Lathyrus
sativus L. Indian J. Genet. 61:53-56

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Effect of Gamma Rays on Induction of Chlorophyll

Mutants in Ginger Genotypes (Zingiber Officinale Rosc.)

109

APPENDICES
Table 1: Frequency of Chlorophyll Mutants (%) of Ginger Genotypes in VM2 Generation
Genotypes

Rio-de-Janeiro
G1

Maran
G2

Treatments
T1(3 Gy)
T2(4 Gy)
T3(5 Gy)
T4(6 Gy)
T5(7 Gy)
T6(8 Gy)
T7(9 Gy)
T8(10 Gy)
T9(11 Gy)
Mean
T1(3 Gy)
T2(4 Gy)
T3(5 Gy)
T4(6 Gy)
T5(7 Gy)
T6(8 Gy)
T7(9 Gy)
T8(10 Gy)
T9(11 Gy)
Mean
Grand Mean

Total M2 Plants
Studied
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50

Total Number of Chlorophyll

Mutants
1.35
1.57
2.21
2.57
1.00
2.00
1.00
1.30
1.00
1.57
2.47
2.00
1.00
1.00
1.00
1.12
1.21

Mutation
Frequency (%)
2.70
3.14
4.42
5.14
2.00
4.00
2.00
2.60
2.00
3.14
4.94
4.00
2.00
2.00
2.00
2.23
2.42

Table 2: Types of Chlorophyll Mutants of Ginger Genotypes in VM2 Generation

Total
Relative Percentage of Chlorophyll Mutant
Number of
Genotypes
Treatments
Chlorophyll
Xantha
Albina
Chlorina
Straita
Mutants
T1(3 Gy)
1.35
0.50 (37.04)
0.43 (31.85)
0.42 (26.67)
T2(4 Gy)
1.57
1.00 (63.69)
0.29 (18.47)
0.28 (17.83)
T3(5 Gy)
2.21
1.00 (45.25)
0.33 (14.93)
0.44 (19.91)
0.44 (19.91)
T4(6 Gy)
2.57
1.50 (58.37)
0.57 (22.18)
0.50 (19.46)
Rio-deT5(7 Gy)
1.00
0.50 (50.00)
0.50 (50.00)
Janeiro
T6(8 Gy)
2.00
1.00 (50.00)
0.50 (25.00)
0.50 (25.00)
G1
T7(9 Gy)
1.00
0.50 (50.00)
0.50 (50.00)
T8(10 Gy)
T9(11 Gy)
Mean
1.30
0.44 (34.19)
0.37 (28.46)
0.23 (17.86)
0.25 (18.97)
T1(3 Gy)
1.00
0.50 (50.00)
0.50 (50.00)
T2(4 Gy)
1.57
1.00 (63.69)
0.57 (36.31)
T3(5 Gy)
2.47
1.50 (60.73)
0.50 (20.24)
0.47 (19.03)
T4(6 Gy)
2.00
0.50 (25.00)
1.00 (50.00)
0.50 (25.00)
T5(7 Gy)
1.00
0.50 (50.00)
0.50 (50.00)
Maran
T6(8 Gy)
1.00
0.75 (75.00)
0.25 (25.00)
G2
T7(9 Gy)
1.00
0.25 (25.00)
0.75 (75.00)
T8(10 Gy)
T9(11 Gy)
Mean
1.12
0.39 (34.86)
0.17 (14.94)
0.45 (40.54)
0.11 (9.66)
Grand mean
1.21
0.42 (34.50)
0.27 (22.22)
0.34 (28.33)
0.18 (14.67)
Figures in the parenthesis indicate arc sine transformed values.

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P. Prasanna Kumar, S. Subramanian, J. Suresh, J. R. Kannan Bapu & R. Gnanam

Figure 1: Frequency of Chlorophyll Mutants (%) of Ginger Genotypes in Vm2 Generation

a. Xantha

c. Albina

b. Chlorina

d. Striata

Figure 2: Effect of Gamma Rays on Induced Chlorophyll Mutants in VM2 Generation

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