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Research Article
 

Evaluation of Mutant Generations of Irradiated Soybean (Glycine max (L.) Merrill) in the Guinea Savannah Agroecology of Ghana



I.K. Addai, A. Bawa and K. Asamoah Jnr
 
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ABSTRACT

Background and Objective: Gamma rays at different doses have been used to induce mutation in several species. The objective of the study was to determine mutant lines with improved agronomic traits. Materials and Methods: Seeds of soybean variety ‘Jenguma’ were irradiated with gamma rays at 150, 200, 250 and 300 Gy using un-irradiated (0 Gy) as control. The M2 seeds were screened and the promising lines were selected and were advanced to M3 generation. Desired M3 mutants were evaluated for improved agronomic traits in RCBD in the Guinea Savannah Agro-ecology. Results: The irradiation was found to have had great potential to improve the yield and other important agronomic traits for the crop. Desired characteristics were found with 200, 250 and 150 Gy treated seeds. Maturity periods for seeds selected were shorter. The shattering resistance of the selected M2 seeds of 200 and 250 Gy were also found to be a potential improvement over the parental variety ‘Jenguma’ which was originally bred to control pod shattering. More desired traits were found in the 200 and 250 Gy mutant lines with only a few in the 150 and 300 Gy. Conclusion: Promising mutant genotypes should be tested in multi-locational trials to determine their suitability in the various agro-ecologies for release as varieties and production by farmers in these areas.

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I.K. Addai, A. Bawa and K. Asamoah Jnr, 2022. Evaluation of Mutant Generations of Irradiated Soybean (Glycine max (L.) Merrill) in the Guinea Savannah Agroecology of Ghana. International Journal of Plant Breeding and Genetics, 16: 1-9.

DOI: 10.3923/ijpbg.2022.1.9

URL: https://scialert.net/abstract/?doi=ijpbg.2022.1.9
 

INTRODUCTION

Soybean (Glycine max (L.) Merrill) is an annual legume crop belonging to the botanical family Fabaceae. It is an economically important leguminous crop worldwide and also the most important legume in Ghana1. Among soybean growing regions in Ghana, the largest production occurs in the northern part of Ghana, which lies within the Guinea Savannah and Sahel agro-ecological zones2. Soybean is gaining prominence in Ghana mainly because of its multipurpose usage3. In Ghana, soy cake is an excellent source of protein feed for the livestock industry4. Soybean serves as a good source of unsaturated fatty acids, minerals (Ca and P) and vitamins A, B, C and D5. It is also used to fortify various traditional foods such as gari, sauce, stew, soup, banku and kenkey to improve their nutritional levels4. Soybean research and production in Ghana are besieged with many constraints. The use of unimproved low yielding varieties, poor seed viability, low research effort, pests and disease infestation, the narrow genetic base of cultivars and pod shattering among others continue to bedevil production of this crop. As a result, high soybean production is not obtained and there is a big gap between what is currently produced and what is needed. As a way of improving the production level, one of the major areas of research in the country would be the development of high yielding varieties that are also resistant to pod shattering.

Introducing new improved soybean genotypes may increase genetic diversity, which would facilitate the development of new varieties that can address some of the constraints in soybean production6. The production of soybean crops with new desirable traits has recently been achieved through genetic engineering. However, health, religious, social and ethical considerations concerning the release of transgenic crops to the environment remains a course for discussion in the country. Mutagenesis could be one of the key solutions to numerous constraints facing soybean production and agriculture in Ghana. Mutagenic agents, either physical or chemical, can be used to induce mutations and generate genetic variations from which desired mutants would be selected. In the present study, M2 seeds were screened and the selected promising lines advanced to M3 generation. The desired M3 mutants were evaluated for improved agronomic traits. The study therefore aimed at determining the effectiveness of selection in the M2 and M3 generations with the view to comparing the progress made in the selection of desirable agronomic traits in the 2 generations.

MATERIALS AND METHODS

Experimental site: The researches were conducted at the experimental field of the University for Development Studies (UDS) in the Faculty of Agriculture, Nyankpala campus in the Northern Region of Ghana. Nyankpala is located about 16 km west of Tamale Metropolis and lies on the latitude 09°24’15.9’’N and longitudes 01°00’12.1’W and an altitude of 183 M (msl) of the interior Guinea Savannah agro-ecological zone of Ghana. The studies were carried out during the cropping seasons of 2015, 2016 and 2017, respectively. The experimental site is characterized by tree and weed species such as Neem tree (Azadirachta indica), Shea tree (Vitellaria paradoxa), Dawadawa tree (Parkia biglobosa), Teak (Tectona grandis), Broom weed (Sida acuta), Speargrass (Imperata cylindrica L. ), Pigweed (Boerhavia diffusa) Andropogon gayanus7. The soil in the area is Nyankpala series and is made up of mainly sand and loam. Rainfall in the experimental area starts in late April and reaches a peak in July-September, there is a sharp decline and little or no rain in November and December8. The area experiences total precipitation of about 1,100 mm per annum, with a range from about 800-1500 mm (Table 1). The experimental area has an average ambient temperature usually high all year round (about 28°C) but the harmattan months of December and January, are characterized by the minimum temperature that may fall to 13°C at night, while March and April, may experience 40°C in the early afternoon (Table 2). The experimental site also has an average Relative Humidity (RH) of 64 during May-December, (Table 3).

Land preparation, experimental design and crop husbandry: The field was ploughed and harrowed by a tractor and later manually levelled, ridged and demarcated according to the planting distance of 50×20 cm for soybean in the various cropping seasons. This was done with the aid of tape measure, lines and pegs. A plot size measuring 2×2 m, with 1.5 m space between blocks and 1 m space between plots and replication were considered. The trials were laid out using Randomized Complete Block Design (RCBD) with three replications. Soybean variety (Jenguma) were irradiated to generate M1 seeds in 2015 using 150, 200, 250 and 300 Gy doses of gamma irradiation from Cobalt-60 source and planted. Some un-irradiated seeds (0 Gy) were also planted to serve as the control. The harvested M1 seeds were planted to obtain M2 seeds in 2016. Selected M2 seeds were planted and evaluated for improved agro-morphological traits in 2017. Weeding was carried out manually on 2 weeks’ interval basis to control the growth of weeds.

Table 1: Total rainfall and its' distribution during the experimental period
2015
2016
2017
Rainfall (mL)
Total rainfall (mL)
Number of rain days
Total rainfall (mL)
Number of rain days
Total rainfall (mL)
Number of rain days
May
40.0
5
58.7
6
38.4
4
June
158.9
8
157
8
68.9
5
July
222.7
11
236.4
14
146.4
12
August
238.0
14
240.2
14
180.5
15
September
260.9
16
256.7
17
227.5
16
October
132.7
12
133.9
11
124.3
8
November
19.5
1
20
2
7.6
1
December
0
0
0
0
0
0
Total
1072.7
67
1102.9
72
793.6
61
Mean
134.1
8.4
137.9
9.0
99.2
7.6


Table 2: Temperature distribution during the experimental period
2015
2016
2017
Temperature (°C)
Minimum
Maximum
Mean
Minimum
Maximum
Mean
Minimum
Maximum
Mean
May
25.3
34.3
29.8
26.8
35.5
31.2
25.9
35.5
30.7
June
24.3
31.1
27.8
25.6
33.2
29.4
25.3
34.3
29.8
July
23.6
29.7
27.8
24.9
30.9
27.9
24.2
30.7
27.5
August
22.9
28.9
26.7
23.3
30.4
26.9
24
29.8
30.8
September
22.6
30.2
25.9
23.1
30.5
26.8
24
27.5
27.4
October
23.2
32
26.4
23.5
32.7
28.1
24.5
32.5
28.5
November
22.9
35
29
24.3
35.6
30
23.2
35.7
29.5
December
19.6
35.5
27.5
20.7
35.8
28.4
18.6
34.2
26.4
Total
184.4
256.7
220.9
192.2
264.6
228.7
189.7
260.2
230.6
Mean
23.0
32.0
27.6
24.0
33.0
28.5
23.7
32.5
28.8


Table 3: Relative humidity distribution during the experimental period
2015
2016
2017
Relative humidity (RH)
Minimum
Maximum
Mean
Minimum
Maximum
Mean
Minimum
Maximum
Mean
May
60
83
73
56
83
70
67
85
76
June
68
89
79
64
88
70
48
94
69
July
75
93
84
71
91
81
56
94
75
August
75
93
84
73
93
83
64
96
80
September
74
89
79
75
93
84
72
94
83
October
68
90
79
68
90
79
67
93
80
November
53
82
68
54
82
68
39
74
57
December
52
68
60
38
56
44
37
57
44
Total
525
687
606
499
676
579
450
687
564
Mean
65.6
85.8
75.7
62.3
84.5
72.3
56.2
85.8
70.5

Data collection and analysis: Data generated from 2015-2017 from the various generations (plant height, chlorophyll content, number of branches, percentage pod shattering, number of days to 50% flowering, number of days to maturity, number of seeds per pod, number of pods per plant, pod length, 100 seed weight and total grain yield) were subjected to combined analysis for variation in factorial experiments (factor 1 being mutant genotypes and factor 2 being mutant generations) in RCBD. Genstat (18 edition) statistical package was used in the analysis. Means were separated using the Least Significant Difference test (LSD) at a 5% probability level.

RESULTS

Number of secondary branches: Irradiation dose and generation effect as well as their interactions significantly (p<0.05), influenced the number of branches. The 150 Gy mutant lines recorded the highest number of branches, while the 250 Gy mutant lines recorded the lowest number of branches (Fig. 2a). The M3 generation in general recorded the highest number of branches whilst plants from the M1 generation recorded the lowest number of branches (Fig. 2b). However, plants irradiated with 150 Gy recorded the highest number of branches at M2 whilst those treated with 200 Gy dose recorded their highest number of branches at M3 (Fig. 2c).

Image for - Evaluation of Mutant Generations of Irradiated Soybean (Glycine max (L.) Merrill) in the Guinea Savannah Agroecology of Ghana
Fig. 1(a-b): Influence of (a) Irradiation dose and (b) Generations on plant height of soybean (Glycine max (L.) Merrill) mutants during field experimentation
Error bars represent Mean±SEM


Image for - Evaluation of Mutant Generations of Irradiated Soybean (Glycine max (L.) Merrill) in the Guinea Savannah Agroecology of Ghana
Fig. 2(a-c): Influence of (a) Irradiation, (b) Generations and (c) Genotype×generation on the number of branches of soybean (Glycine max (L.) Merrill) mutants
Error bars represent Mean±SEM

Days to 50% flowering: Treatments with gamma irradiation doses and generation effect as well as the interaction between the 2 factors significantly affected (p<0.05) number of days to 50% flowering. Plants from 150, 200 and 250 Gy recorded earliness in terms of flowering (Fig. 3a), whilst plants from the M2 and M3 showed the same number of days to 50% flowering (Fig. 3b). Plants from 150, 200 and 250 Gy had a similar number of days to 50% flowering at the M2 and M3 generations (Fig. 3c).

Number of days to maturity: The number of days to maturity followed a similar trend as the number of days to 50% flowering (Fig. 4a-c).

Pod shattering: Pod shattering was highly significant (p<0.001) among dosages and also single effect of generation and the interaction between the 2 factors had a significant (p<0.05) influence on pod shattering. The 0 Gy application dose recorded the highest number of pod-shattering, whilst the 250 Gy dose recorded the least number of pod-shattering (Fig. 5a). The M1 generation recorded the highest number of pod-shattering per plant, whilst the M2 generation recorded the least (Fig. 5b). The M1 generation at 0 and 150 Gy recorded the highest pod shattering, followed by M3 and M2 at 0 Gy. Plants from the 200 Gy in all generations showed resistance to pod shattering (Fig. 5c).

Total grain and components of yield: Plants that were irradiated with 200 Gy of gamma-ray recorded relatively higher grain yield and components of yield especially at the M2 generation (Table 4).

Image for - Evaluation of Mutant Generations of Irradiated Soybean (Glycine max (L.) Merrill) in the Guinea Savannah Agroecology of Ghana
Fig. 3(a-c): Influence of (a) Irradiation, (b) Generations and (c) genotype×generation on the number of days to 50% flowering of soybean (Glycine max (L.) Merrill) mutants
Error bars represent Mean±SEM


Image for - Evaluation of Mutant Generations of Irradiated Soybean (Glycine max (L.) Merrill) in the Guinea Savannah Agroecology of Ghana
Fig. 4(a-c): Influence of (a) Irradiation, (b) Generations and (c) Genotype×generation on the number of days to maturity of soybean (Glycine max (L.) Merrill) mutants
Error bars represent Mean±SEM


Image for - Evaluation of Mutant Generations of Irradiated Soybean (Glycine max (L.) Merrill) in the Guinea Savannah Agroecology of Ghana
Fig. 5(a-c): Influence of (a) Irradiation, (b) Generations and (c) Genotype x generation on pod-shattering of soybean (Glycine max (L.) Merrill) mutants
Error bars represent Mean±SEM


Table 4: Grain yield and yield components of mutant lines of soybean (Glycine max (L.) Merrill) planted in Nyankpala for improved growth and yield
Dosage
Grain yield (t ha‾1)
100-seed weight (g)
Number of pods per plant
Pod length (cm)
0
1.76
28.27
68
2.5
150
1.27
30.74
60
2.8
200
2.06
29.89
55
2.9
250
1.76
30.26
68
2.4
300
1.81
29.17
60
2.2
LSD (0.05)
3.56
2.53
9.29
0.11
Generation
M1
0.99
36.03
15
2.5
M2
2.48
33.66
50
2.4
M3
1.73
19.31
106
2.9
LSD (0.05)
0.75
1.96
7.19
0.23

DISCUSSION

There was significant variation among the irradiation doses, mutant generations and their interaction for grain yield and other agronomic traits measured. Gamma irradiation and selection significantly affected plant height. This agrees with the findings of the previous studies9,10, who reported a reduction in height of basmati rice following gamma irradiation. The un-irradiated plants initially grew very slowly but later became the tallest, flowering lately and reaching maturity lately whilst plants irradiated with 150 and 200 Gy stopped growing tall and produced more branches. The biological effect of gamma-rays is based on the interaction with atoms or molecules in the cell11. Induced mutation using gamma irradiation might have resulted in retardation of growth. The observation made here is similar to those researchers12,13, who also observed the effect of high dose treatment on plant derivatives of M1 mungbean (Vigna radiata (L.) Wilczek), where plant height decreased due to the treatment of gamma irradiation. Gamma irradiation treatment increased the average plant height compared with the unirradiated controls14. Many studies have shown that treatment with a higher dose of gamma rays was inhibitory, whereas lower exposures were sometimes stimulatory15,16. This could be due to mutagenic induced chromosomal damage during cell division17. Gamma irradiation disturbs the synthesis of protein, hormone balance, leaf gas exchange, water exchange and enzyme activity18. However, previously19 reported an increased plant height in soybean due to gamma rays. A similar observation was made in the M3 plants that were initially treated with a 300 Gy dose of gamma-ray because these plants outgrew the 200 and 250 Gy treated plants. In-plant production, reduction in height may be advantageous as short plants generally resist lodging and hence avoid yield losses that may be associated with it. A reduction in plant height has also been reported, following the application of 250 and 200 Gy in soybeans20,21. The height difference was therefore more distinct in the M3 and M2 populations.

The number of days to (50%) flowering varied significantly among the generations of mutants of soybean. The difference was more distinct in 200 and 250 Gy mutants, which took about half the number of days taken by the 0 Gy. Zaka et al.22 had earlier reported a reduction in the number of days to flowering in pea (Pisum sativum) following irradiation. Early maturity was observed in the soybean lines especially mutants from 150, 200 and 250 Gy. The 0 Gy took almost the same number of days as 300 Gy to mature. Ranjitha et al.23 also selected mutants from gamma-irradiated soybean that proved to be early maturing and dwarf stature. Pod shattering is the opening of pods along both the dorsal and ventral sutures of the soybean pod. It is essential for the propagation of wild plant species bearing seeds in pods but is a major cause of yield loss in legume and crucifer crops. According to Valkama et al.24, the gene for the shattering-resistant genotype, pdh1, is defective in soybean, having a premature stop codon. In this study, tolerance to pod shattering varied significantly among genotypes and was high in plants treated with 200 and 250 Gy. Ionizing radiation affects cellular components, thus, potentially inducing physiological changes in plants. This may be the mechanism that prevents the pod from shattering. The number of pods per plant differed significantly among the treatments. The result in part contradicts the findings of Pasupuleti et al.25, who reported that the number of pods per plant decreased in all the genotypes (of a black gram) as the dose of irradiation increased. But Rahman et al.26 observed that the treatment with gamma rays stimulated most of the quantitative characters of Vigna unguiculata such as number of branches, number of clusters, number of pods and yield. Khan and Goyal27, also reported more number of pods per plant in mung beans treated with gamma rays. Hundred seed weight did not differ significantly among the treatments. A study of M1 of cowpea, groundnut and soybean using gamma-ray at the same dose reported no significant differences28. Since hundred seeds weight did not significantly differ among the treatments, the number of pods that had a significant variation seemed to have caused the significant differences in the yields of soybean.

The number of pods is a good selection criterion for increasing yield in grain legumes. This finding corroborates with the observation made by Rahimi and Bahrani29, who reported the highest grain yield increase in canola irradiated with 0.2 kGy of gamma rays. Similarly, they also reported a significant improvement of 1000 kernel weight and harvest index for 0.1 kGy gamma-ray treatments. The sizes of grains in the mutants were comparably high as compared to the parent and were selected for high yield. The M2 generation recorded the highest yield compared to the other generations. The results of the M3 generation showed that it is possible to effectively increase grain yields components at a dose of 200 and 250 Gy. Similar observations were also made in other plants like black gram, cowpea and sesame30. Improvement of agronomic characteristics by using gamma radiation has also been reported in several studies. A study revealed a significant increase in chickpea grain yield using gamma irradiation at 0.6 kGy31. An increase has also been observed, in the number of pods per plant in all the varieties for gamma irradiation at 0.2 and 0.4 kGy doses32. However, Khan et al.31, reported a decrease in pod number at 0.4 kGy treatment and an increase at 50 kGy without a change in the number of seeds per pod. The selected seeds especially of 200 and 250 Gy mutant lines should be advanced for multi-locational trials to determine the suitability of the selected mutants in various agro-ecologies for release as varieties for production by farmers.

CONCLUSION

The study showed that plants treated with 200 and 250 Gy were found to be highly resistant to shattering than the un-irradiated (0 Gy/control), believed to be an improved variety against field shattering. Field performance (agronomic traits) of soybean plants treated with 200, 250, 150 and 300 Gy were on the average higher than the parental line (control/0 Gy). These mutant lines also had shorter stems as compared to the parent line, which has an advantage in terms of lodging during heavy rains and strong winds. Yield and yield components results for soybean were higher among the M2 populations followed by M3 while M1 had the least. Individual plants with desired characteristics than the parental variety ‘Jenguma’ were selected for multi-locational trials in the next generation. The number of pods per plant, seed size and seed weight were particularly considered during selection.

SIGNIFICANCE STATEMENT

This study discovered that mutagenesis could help improve yield, growth parameters and pod-shattering resistance of soya beans. Through this study, the researcher had discovered that plants treated with 200 and 250 Gy were found to produce more grain yield, improved growth parameters and were highly resistant to pod-shattering than the un-irradiated (0 Gy/control). Therefore, farmers who use these mutant lines are likely to get a better grain yield. This will result in increased soybean productivity in the Guinea Savannah Agroecology of Ghana.

ACKNOWLEDGMENT

The authors of this study are most grateful to the Faculty of Agriculture of UDS for providing funds for land preparation before planting. Sincere thanks also go to all technicians of the Department of Agronomy, UDS for their help in data collection, weeding and other cultural practices.

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