Short Communication | Volume 11, Issue 3, May, 2023

Effects of gibberellic acid on seed dormancy of black gram (Vigna mungo L.)

Vinothini Nedunchezhiyan Mohanasundari Palanivel P. A. Akhila Jabeen Poovarasan Thangavel Bhavyasree Ramakrishnan Manonmani Velusamy Sakila Muthusamy Isong Abasianyanga Edm   

Open Access   

Published:  Apr 04, 2023

DOI: 10.7324/JABB.2023.45506
Abstract

Black gram or Urdbean (Vigna mungo) is a widely cultivated pulse crop in India. The seeds of this crop show dormancy and are not able to germinate as and when required. Investigation of the effect of gibberellic acid (GA3) in breaking the dormancy of black gram seeds is the objective of this experiment. To break the dormancy, seeds were soaked with different concentrations of GA3 along with hydro-primed seeds and control seeds for 3 h and shade dried. Evaluation of germination and seedling growth of these primed seeds under controlled environment conditions revealed that the seed treated with 350 ppm of GA3 observed in maximum physiological parameters. Hence, the treatment of seeds with 350 ppm of GA3 is an effective dormancy-breaking treatment in black gram.


Keyword:     Black gram Hard seeds Dormancy Gibberellic acid Germination


Citation:

Nedunchezhiyan V, Palanivel M, Jabeen PAA, Thangavel P, Ramakrishnan B, Velusamy M, Muthusamy S, Edm IA. Effects of gibberellic acid on seed dormancy of black gram (Vigna mungo L). J App Biol Biotech. 2023;11(3): 256-259. https://doi.org/10.7324/JABB.2023.45506

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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1. INTRODUCTION

Black gram belonging to Fabaceae family is on among the major pulse crops in area and productivity. Black grams have a high nutritive value containing 20–25% proteins, 40–47% starch, and other essential vitamins and carbohydrates. Major constraints during seed testing and sowing of Vigna species are the presence of hard seeds [1].

Leguminous seeds have high dormancy due to the seed structure. Black gram has seed dormancy of about 3–4 months. The classification of dormancy [2] five classes of dormancy based on the embryo or other components of the seed. Dormancy occurring in pulses is physical dormancy, the frequency of physical dormancy increases with the environment [3]. Black gram has only physical dormancy due to water-impermeable layers within the seed coat [4]. When the seeds are exposed to water the seeds that cannot absorb the water due to the impermeable layer, absorption of water is possible only after the removal of the impermeable layer by seed scarification. In black gram, due to the seed dormancy, the germination percentage is less and there will be irregular seed germination and establishment, leading to the subsequent yield loss.

Increased plant establishment and increase in yield productivity are highly related to seed quality. Various technologies are involved in increasing germination, seedling growth, and productivity. Seed priming is one of the techniques, which is highly effective, inexpensive and of low risk [5]. Pre-sowing seed treatment priming includes soaking of seeds in water, chemicals, micronutrients, biocontrol agents, or bio-fertilizers, allowing the metabolic actions before germination in the seeds; hence, there is reduction in the germination time and increase in the efficiency of seedling establishment increasing the germination percentage of seeds [6].

In the chemical treatments to improve seed germination and establishment, seeds are treated with growth regulators like gibberellic acid (GA3) [7]. GA3 is a natural regulator highly influencing the physiological parameters of plants and, hence, has multiple uses in the agriculture and horticulture industry. The important impact of GA3 is the enhancement of seed germination, which has been demonstrated in ornamental plants [8]. During germination, GA3 is released from the embryo hence stimulating mRNA and production of a-amylase [9].

Improving the seed germination and seedling growth by breaking the seed dormancy can be done by seed priming technique, using different solutions when imbibed by the seed improves the seed germination. Several seed priming agents such as potassium nitrate and phytohormone like GA3 can be used for effective priming of seeds [10]. The results of GA3 treatments on dormant black gram seeds and their effects on overcoming the seed dormancy are discussed in the paper.


2. MATERIALS AND METHODS

2.1. Collection of Materials

The research was conducted to evaluate the effect of GA3 on dormancy breaking and seedling growth on the black gram at “SRM College of Agricultural Sciences, SRM Institute of Science and Technology.” The source material for the study genetically pure black gram seeds variety ADT 6 was collected from farm office at Tamil Nadu Agricultural University, Coimbatore.

2.2. Treatments

Freshly harvested black gram seeds were used for the study. Initially, a 1000 ppm GA3 solution was prepared to make different concentrations for treatments. Presoaking of seeds in different concentrations of GA3 solution, that is, 50, 100, 150, 200, 250, 300, 350, 400, 450, and 500 ppm and without any treatment as control. Seeds soaked in the GA3 for 3 h, then shade dried under room temperature. The physiological growth parameters were analyzed in the treated and control plants which were grown under controlled conditions.

2.3. Germination %

Four hundred seeds used for conducting the seed germination test by roll towel method [11], the temperature and RH maintained at 25 ± 2°C and 95 ± 2%, respectively. At end of the test on 7th day, the total number of normal seedlings was counted and expressed in percentage.

2.4. Root Length (cm)

At the final count, ten normal seedlings were selected from each treatment, for measuring root length and removing the plant intact with the entire root system. Between the collar and the tip of the root, the root’s length was measured, and its mean was given in centimeters.

2.5. Shoot Length (cm)

Measure the shoot length with same seedlings used for root length measurement. The shoot length was recorded length between the collar and tip of the primary leaf and their mean was reported in centimeter.

2.6. Vigor Index

The following formula was used to determine the vigor index, and the mean values were expressed as whole numbers [12].

“Vigour index = Germination percentage × Total seedling length (cm)”

“[Total seedling length = Root length (cm) + Shoot length (cm)]”

2.7. Statistical Analysis

Completely randomized block design was used for the experiment, and there were four replications.


3. RESULTS AND DISCUSSION

Black gram seeds were subjected to GA3 treatments with different concentrations and recorded seed quality parameters. All physiological measures, including seed germination per cent, seedling lengths, and vigor index, were significantly affected by the treatments. The seeds primed with 350 ppm of GA3 recorded higher germination per cent (88%) which was on par with 400 ppm and 450 ppm of GA3, and was significantly greater than the control which showed the lowest germination percentage with 26% [Figure 1]. From the outcomes of the experiments, the germination and growth of dormant black gram seeds are significantly influenced by the GA3. In the study, GA3 had significant effect of two functions on the seed germination. Initially, GA3 increases the growing potential of embryo hence promoting the germination. Later, GA3 plays significant role in overcoming the mechanical constrains due to hard seed coats by weakening the tissues around the radicle [13].

Figure 1: Effect on gibberellic acid (GA3) treatments on germination percentage of black gram. Treatment details: T1 – Control, T2 – 50 ppm, T3 – 100 ppm, T4 – 150 ppm, T5 – 200 ppm, T6 – 250 ppm, T7 – 300 ppm, T8 – 350 ppm, T9 – 400 ppm, T10 – 450 ppm, and T11 – 500 ppm.



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During the exogenous application of gibberellins, their main function is to compensate for the inhibition caused by abscisic acid and while endogenous application, there will be an increase in GA3 production playing a major role in seed germination. GA3 has a major role in overcoming dormancy and controlling the hydrolysis of reserves. The amount of GA3 present stimulates the synthesis, activation, and secretion of hydrolytic enzymes, primarily-amylase, which releases reducing sugars and amino acids necessary for embryo growth [14].

Black gram ADT 6 variety treated with 300 ppm GA3 increased the germination and was found effective in breaking the dormancy compared to other seeds. According to the results of [15], the same treatment improved germination percentage and vigor index in lentil seeds. With the increased germination percentage, while soaking the seeds in GA3, there is the stimulation of cytological enzymes that produce the enzyme α–amylase; hence, there is the fast conversion of insoluble starch into soluble sugars initiating root and shoot development, removing the barriers during metabolism [16]. GA3 can leach out the inhibitors in germination, in turn, breaking the dormancy of seeds. The role of GA3 in leaching out inhibitors is considered an important role in germination [17,18].

GA3 at different concentrations shows increased germination percentage as it induces the production of mRNA molecules that codes for hydrolytic enzymes in the germinating seeds [19]. Degradation of food reserves accumulated in the endosperm is degraded by the hydrolytic enzymes mainly amylase and protease. The energy and nourishment for proper seed germination are attained from the degradation of carbohydrates and proteins in the endosperm [20]. The results prove the findings of [21], the required conditions for absorption of water and cell growth, like an increase in the cell size is done by GA3, by the release and transition of calcium to the cytoplasm from the cell wall. After the cell growth, calcium returns to the cell wall and stiffens the cell structure. By the absorption of water by seeds, GA3 is produced by the embryo inducing the synthesis of α- and β-amylase the hydrolytic enzyme in the aleurone layer involved in the conversion of starch and glucose which are absorbed by the embryo. In the endosperm, GA3 increases DNA replication and dietary material analysis by acting on the proteins that make mRNA [22].

The root’s length was measured in centimeters from the collar to the tip, with the mean length being recorded and used as the root length. From the observations, the best result was observed in the concentration of 300 ppm and 350 ppm which expressed a root length of 15.2 cm of GA3 considering the control which expressed the lowest root length of 9.3 cm [Figure 2]. The length from base of the seedlings to the top most leaf in each seedling was measured in centimeter and the mean length recorded taken as the shoot length. The longest shoot length was 26.6 cm observed in seeds primed with 350 ppm of GA3 as compared to control which expressed the 18.1 cm shortest shoot length [Figure 3]. When seeds are treated with GA3, they showed a higher root length of 15.2 cm in 300 ppm and 350 ppm and shoot length of 26.6 cm in 350 ppm. Hence proving the role of GA3 in inducing root and shoot growth by inducing mitotic division in the respective regions of plants [23]. An increase in root and shoot growth by the increased cell division and cell elongation in the cambium region of the internodes is due to the stimulation of GA3 [24]. GA3 treatment activated the dormant embryo in the seed and improved the shoot growth by increasing the cell division, elongation, and multiplication, which is proved by the increased seedling length. The result coincides with the results of [25,26].

Figure 2: Effect on GA3 treatments on root length (cm) of black gram. Treatment details: T1 – Control, T2 – 50 ppm, T3 – 100 ppm, T4 – 150 ppm, T5 – 200 ppm, T6 – 250 ppm, T7 – 300 ppm, T8 – 350 ppm, T9 – 400 ppm, T10 – 450 ppm, and T11 – 500 ppm.



[Click here to view]
Figure 3: Effect on GA3 treatments on shoot length (cm) of black gram. Treatment details: T1 – Control, T2 – 50 ppm, T3 – 100 ppm, T4 – 150 ppm, T5 – 200 ppm, T6 – 250 ppm, T7 – 300 ppm, T8 – 350 ppm, T9 – 400 ppm, T10 – 450 ppm, and T11 – 500 ppm.



[Click here to view]

While comparing the calculated values of the vigor index with that of the control, the seeds treated with 350 ppm of GA3 exhibited the highest value of vigor index of 3678, compared to the control which expressed the lowest vigor index of 712 [Figure 4]. GA3-treated seed shows maximum vigor index of about of 2941 at 400 ppm. This result agrees with the findings of [16]. Priming has a great impact on seed germination, and seedling length. GA3 is present in low concentrations in plants by its high impact in the growth and development of plants. The application of GA3 induces positive impacts when applied at adequate quantities, when the concentration increases that there is an adverse effect on the plant. The germination percentage and seedling length have a direct relation to the vigor index of the seed. Hence, higher vigor index of GA3-treated seeds is due to the influence of GA3 on the germination and seedling length in chickpeas in the laboratory [27]. Therefore, it can be concluded that seeds treated with 350 ppm GA3 were found to be most effective and could be recommended for breaking the dormancy of black gram seeds.

Figure 4: Effect on GA3 treatments on vigor index of black gram. Treatment details: T1 – Control, T2 – 50 ppm, T3 -100 ppm, T4 - 150 ppm, T5 – 200 ppm, T6 – 250 ppm, T7 – 300 pp m, T8 – 350 ppm, T9 – 400 ppm, T10 – 450 ppm, and T11 – 500 ppm.



[Click here to view]

4. CONCLUSION

The present study revealed that seeds treated with 350 ppm of GA3 showed maximum physiological growth parameters, namely, seed germination per cent, seedling length, and higher vigor index. Hence, the priming with GA3 at a rate of 350 ppm can be effectively used as a pre-germinative treatment to break the dormancy in black gram.


5. AUTHORS’ CONTRIBUTIONS

All authors made substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data; took part in drafting the article or revising it critically for important intellectual content; agreed to submit to the current journal; gave final approval of the version to be published; and agreed to be accountable for all aspects of the work. All the authors are eligible to be an author as per the International Committee of Medical Journal Editors (ICMJE) requirements/guidelines.


6. FUNDING

This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.


7. CONFLICTS OF INTEREST

The authors report no financial or any other conflicts of interest in this work.


8. ETHICAL APPROVALS

Ethics approval was not required for this study.


9. DATA AVAILABILITY

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.


10. PUBLISHER’S NOTE

This journal remains neutral with regard to jurisdictional claims in published institutional affiliation.

REFERENCES

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2.  Baskin JM, Baskin C. The great diversity in kinds of seed dormancy:A revision of the Nikolaeva-Baskin classification system for primary seed dormancy. Seed Sci Res 2021;31:1-29. [CrossRef]

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5.  Thakur M, Tiwari S, Kataria S, Anand A. Recent advances in seed priming strategies for enhancing planting value of vegetable seeds. Sci Hortic 2022;305:111355. [CrossRef]

6.  Abiri R, Shaharuddin NA, Maziah M, Yusof ZN, Atabaki N, Sahebi M, et al. Quantitative assessment of indica rice germination to hydropriming, hormonal priming and polyethylene glycol priming. Chil J Agric Res 2016;76:392-400. [CrossRef]

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10.  Bhavyasree RK, Vinothini N. Enhancement of seed quality through orgopriming in brinjal (Solanum melongena L.). Int J Chem Stud 2019;7:242-4.

11.  International Seed Testing Association. International Rules for Seed Testing. Bassersdorf (Switzerland):International Seed Testing Association;2012.

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16.  Nedunchezhiyan V, Velusamy M, Subburamu K. Seed priming to mitigate the impact of elevated carbon dioxide associated temperature stress on germination in rice (Oryza sativa L.). Arch Agron Soil Sci 2020;1:83-95. [CrossRef]

17.  Bilalis DJ, Katsenios N, Efthimiadou A, Karkanis A, Khah EM, Mitsis T. Magnetic field pre-sowing treatment as an organic friendly technique to promote plant growth and chemical elements accumulation in early stages of cotton. Aust J Crop Sci 2013;7:46-50.

18.  Anburani A, Shakila A. Influence of Seed Treatment on the Enhancement of Germination and Seedling Vigour of Papaya. vol. 851. Almeria (Spain):II International Symposium on Papaya;2008. 295-8. [CrossRef]

19.  Gupta, R, Chakrabarty SK. Gibberellic acid in plant:Still a mystery unresolved. Plant Signal Behav 2013;9:e25504. [CrossRef]

20.  Hota SN, Karna AK, Jain PK, Dakhad B. Effect of gibberellic acid on germination, growth and survival of jamun (Syzygium cumini L. Skeels). J Pharm Innov2018;7:323-6.

21.  Dzayi FH, Rahman. Effect of GA3 and Soaking Time on Seed Germination and Seedling Growth Lemon (Citrus limon L.) High Diploma (Doctoral dissertation, Thesis, college Agriculture University of Salahaddin-Erbil);2010.

22.  Lahuti M, Zare-hasanabadi M, Ahmadian R. Biochemistry and Physiology of Vegetable Hormones. Iran:Ferdosi University Mashhad;2003.

23.  Amri B, Khamassi K, Ali MB, da Silva JA, Kaab LB. Effects of gibberellic acid on the process of organic reserve mobilization in barley grains germinated in the presence of cadmium and molybdenum. S Afr J Bot 2016;106:35-40. [CrossRef]

24.  Chauhan A, AbuAmarah BA, Kumar A, Verma JS, Ghramh HA, Khan KA, et al. Influence of gibberellic acid and different salt concentrations on germination percentage and physiological parameters of oat cultivars. Saudi J Biol Sci 2019;26:1298-304. [CrossRef]

25.  Gawade US. Seed Viability, Germination and Seedling Growth studies in Custard Apple M. Sc.(Ag.) Thesis, Dr. Panjabrao Deshmukhv Krishi Vidyapeeth, AkolaMS (INDIA);2008.

26.  Gholap SV, Dod VN, Bhuyar SA, Bharad SG. Effect of plant growth regulators on seed germination and seedling growth in aonla (Phyllanthus emblica L.) under climatic condition of Akola. Crop Res Hisar 2000;20:546-8.

27.  Poovarasan T, Lakshmi S, Renugadevi J, Senthil A. Seed quality improvement with fruit extracts in blackgram (Vigna mungo L.). J Phytol 2019;11:16-20. https://doi.org/10.25081/jp.2019.v11.3785

Reference

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2. Baskin JM, Baskin C. The great diversity in kinds of seed dormancy: A revision of the Nikolaeva-Baskin classification system for primary seed dormancy. Seed Sci Res 2021;31:1-29. https://doi.org/10.1017/S096025852100026X

3. Nonogaki H. Seed germination and dormancy: The classic story, new puzzles, and evolution. J Integr Plant Biol 2019;61:541-63. https://doi.org/10.1111/jipb.12762

4. Rodrigues-Junior AG, Mello AC, Baskin CC, Baskin JM, Oliveira DM, Garcia QS. A function for the pleurogram in physically dormant seeds. Ann Bot 2019;123:867-76. https://doi.org/10.1093/aob/mcy222

5. Thakur M, Tiwari S, Kataria S, Anand A. Recent advances in seed priming strategies for enhancing planting value of vegetable seeds. Sci Hortic 2022;305:111355. https://doi.org/10.1016/j.scienta.2022.111355

6. Abiri R, Shaharuddin NA, Maziah M, Yusof ZN, Atabaki N, Sahebi M, et al. Quantitative assessment of indica rice germination to hydropriming, hormonal priming and polyethylene glycol priming. Chil J Agric Res 2016;76:392-400. https://doi.org/10.4067/S0718-58392016000400001

7. Camara MC, Vandenberghe LP, Rodrigues C, de Oliveira J, Faulds C, Bertrand E, et al. Current advances in gibberellic acid (GA3) production, patented technologies and potential applications. Planta 2018;248:1049-62. https://doi.org/10.1007/s00425-018-2959-x

8. Joshi K, Kotadiya P, Singh A, Shrimali G, Rami E. Effect of different pre-treatments on seed germination and seedling growth of Adenanthera pavonina. Crop Res 2022;23:229-34. https://doi.org/10.31830/2348-7542.2022.032

9. Blankenship RE. Molecular Mechanisms of Photosynthesis. Hoboken, New Jersey: John Wiley and Sons; 2021.

10. Bhavyasree RK, Vinothini N. Enhancement of seed quality through orgopriming in brinjal (Solanum melongena L.). Int J Chem Stud 2019;7:242-4.

11. International Seed Testing Association. International Rules for Seed Testing. Bassersdorf (Switzerland): International Seed Testing Association; 2012.

12. Abdul-Baki AA, Anderson JD. Vigor determination in soybean seed by multiple criteria. Crop Sci 1973;13:630-3. https://doi.org/10.2135/cropsci1973.0011183X001300060013x

13. Penfield S. Seed dormancy and germination. Curr Biol 2017;27:874-8. https://doi.org/10.1016/j.cub.2017.05.050

14. Han C, Yang P. Studies on the molecular mechanisms of seed germination. Proteomics 2015;10:1671-9. https://doi.org/10.1002/pmic.201400375

15. Pulok MA, Rahman MM, Haque MN, Chakraborty R, Ali MS. Effect of growth regulators on germination and vigor of lentil seeds. J Biosci Agric Res 2015;3:8-14. https://doi.org/10.18801/jbar.030115.26

16. Nedunchezhiyan V, Velusamy M, Subburamu K. Seed priming to mitigate the impact of elevated carbon dioxide associated temperature stress on germination in rice (Oryza sativa L.). Arch Agron Soil Sci 2020;1:83-95. https://doi.org/10.1080/03650340.2019.1599864

17. Bilalis DJ, Katsenios N, Efthimiadou A, Karkanis A, Khah EM, Mitsis T. Magnetic field pre-sowing treatment as an organic friendly technique to promote plant growth and chemical elements accumulation in early stages of cotton. Aust J Crop Sci 2013;7:46-50.

18. Anburani A, Shakila A. Influence of Seed Treatment on the Enhancement of Germination and Seedling Vigour of Papaya. vol. 851. Almeria (Spain): II International Symposium on Papaya; 2008. p. 295-8. https://doi.org/10.17660/ActaHortic.2010.851.45

19. Gupta, R, Chakrabarty SK. Gibberellic acid in plant: Still a mystery unresolved. Plant Signal Behav 2013;9:e25504. https://doi.org/10.4161/psb.25504

20. Hota SN, Karna AK, Jain PK, Dakhad B. Effect of gibberellic acid on germination, growth and survival of jamun (Syzygium cumini L. Skeels). J Pharm Innov 2018;7:323-6.

21. Dzayi FH, Rahman. Effect of GA3 and Soaking Time on Seed Germination and Seedling Growth Lemon (Citrus limon L.) High Diploma (Doctoral dissertation, Thesis, college Agriculture University of Salahaddin-Erbil); 2010.

22. Lahuti M, Zare-hasanabadi M, Ahmadian R. Biochemistry and Physiology of Vegetable Hormones. Iran: Ferdosi University Mashhad; 2003.

23. Amri B, Khamassi K, Ali MB, da Silva JA, Kaab LB. Effects of gibberellic acid on the process of organic reserve mobilization in barley grains germinated in the presence of cadmium and molybdenum. S Afr J Bot 2016;106:35-40. https://doi.org/10.1016/j.sajb.2016.05.007

24. Chauhan A, AbuAmarah BA, Kumar A, Verma JS, Ghramh HA, Khan KA, et al. Influence of gibberellic acid and different salt concentrations on germination percentage and physiological parameters of oat cultivars. Saudi J Biol Sci 2019;26:1298-304. https://doi.org/10.1016/j.sjbs.2019.04.014

25. Gawade US. Seed Viability, Germination and Seedling Growth studies in Custard Apple M. Sc.(Ag.) Thesis, Dr. Panjabrao Deshmukhv Krishi Vidyapeeth, AkolaMS (INDIA); 2008.

26. Gholap SV, Dod VN, Bhuyar SA, Bharad SG. Effect of plant growth regulators on seed germination and seedling growth in aonla (Phyllanthus emblica L.) under climatic condition of Akola. Crop Res Hisar 2000;20:546-8.

27. Poovarasan T, Lakshmi S, Renugadevi J, Senthil A. Seed quality improvement with fruit extracts in blackgram (Vigna mungo L.). J Phytol 2019;11:16-20. https://doi.org/10.25081/jp.2019.v11.3785

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