Legume Research

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Legume Research, volume 45 issue 4 (april 2022) : 435-444

Effect of Seed Priming and Foliar Spraying of PGRs on Morpho-Physiology, Growth and Yield in Green gram (Vigna radiata L.)

R.S. Bhadane1,*, K.R. Prajapati2, K.C. Ombase3, D.B. Patel4
1Oilseeds Research Station, Mahatma Phule Agricultural University, Jalgaon-425 001, Maharashtra, India.
2Vanbandhu Krishi Polytechnic School, S.K. Nagar Agriculture University, Amirgadh-385 130, Gujarat, India.
3Zonal Agricultural Research Station, Mahatma Phule Agricultural University, Solapur-413 006, Maharashtra, India.
4Department of Plant and Physiology, B.A. College of Agriculture, Anand Agricultural University, Anand-388 110, Gujarat, India.
  • Submitted02-06-2020|

  • Accepted13-10-2020|

  • First Online 29-12-2020|

  • doi 10.18805/LR-4434

Cite article:- Bhadane R.S., Prajapati K.R., Ombase K.C., Patel D.B. (2022). Effect of Seed Priming and Foliar Spraying of PGRs on Morpho-Physiology, Growth and Yield in Green gram (Vigna radiata L.) . Legume Research. 45(4): 435-444. doi: 10.18805/LR-4434.
Background: Pre-sowing seed priming and foliar spraying with Plant Growth Regulators is an easy, low cost and low risk technique and also an alternative approach recently used to mitigate the effect of abiotic stresses in agricultural production.

Methods: Seeds of Mung bean var. GAM-5 were primed and also foliar spraying at 30 DAS with CaCl2 2% and 1%, Cycocel 500, 1000 ppm, NAA 25, 50 ppm during summer season of 2015-16 and 2016-17.

Result: The results indicated a significant improvement in morpho-physiological traits, growth parameters, biochemical constituents and thereby yield due to the application of PGRs. Seed priming with 2% CaCl2 followed by 1% foliar spraying at 30 DAS (T11) significantly improved most of morpho-physiological parameters viz., plant height, number of branches per plant, days to 50% flowering, maturity, leaf area, chlorophyll content, seed protein content, number of pods per plant, number of seeds per pod, test weight, pod length, yield per plant, yield per hectare, harvest index in green gram followed by the seed priming with Cycocel 1000 ppm followed by foliar spraying at 30 DAS (T13) and seed priming with NAA 50 ppm followed by foliar spraying at 30 DAS (T15). The treatment T11 was more efficient.
Mung bean (Vigna radiata L.) is an important pulse crop having high nutritive value, suitable for dry land farming and predominantly used as an intercrop with other crops. It contains about 25 per cent protein along with amino acids such as arginine, histidine, lysine and tryptophane etc. It is also considered as a cheap source of protein and other minerals. It has high digestibility and palatability (Singh et al., 2019). It is a very good catch crop in summer and can be grown very well in this season. Mung bean is a short duration, low input requiring crop that matures in 65 to 80 days, photo and thermo-insensitive in nature. However, the productivity of mung bean is low.
       
Efforts made to maximize yield, is largely hampered by adverse effect of abiotic stress such as salinity and drought. These effects cause a huge loss due to low yield and failure of the crop to establish in some cases. Pre-sowing seed priming treatment is simple technique and an alternative approach recently used to overcome the effect of abiotic stresses in agricultural production. It is found to be efficient in improving seed emergence and growth of crops (Sankar Ganesh et al., 2013). It was reported clearly that the hardening treatment enhance seeds vigour by protecting structure of the plasma membrane against injury during stress (Bewley and Black, 1982; Jun Min et al., 2000). It is a well established fact that, pre-soaking seeds with optimal concentration of phytohormones enhance their germination, dry matter accumulation, partitioning and yield of some crop species under condition of environmental stress by increasing nutrient reserves through increased physiological activities and root proliferation (Bozeuk, 1981).
       
Considering the constraints in the production potential of mung bean it is worthwhile to study the influence of different seed hardening and foliar spraying treatments on the production potential of mung bean. It is also of utmost importance to understand the physiological basis of yield attributing characters, partitioning in various plant parts and thereby yield variation due to seed hardening and foliar spraying of various growth regulators and chemicals. The pre-requisite for higher yield is related with the ability of genotype to produce high amount of total dry matter. Patil et al., (2007) reported that the manner in which the net dry matter is produced and distributed among the different parts of plant will determine the economic yield. The present study was therefore, undertaken to assess the morpho- physiological, growth and yield attributing characters in relation to yield in green gram.
The present work was carried out at Agronomy farm, Anand Agricultural University, Anand to study the effect of seed hardening, foliar spraying and their combined effect on morpho-physiology, growth and yield in green gram (Vigna radiata L.) during summer season of 2015-16 and 2016-17. The trial was laid out in a randomized block design with three replications and sixteen treatment combinations including five seed hardening treatments, five foliar spraying treatments, five seed hardening treatments with foliar spraying and one absolute control treatment. Seeds of mung bean var. GAM- 5 were imposed with the following seed treatments.
       
The different solutions of plant growth regulators (PGRs) viz., CaCl2 2%, Cycocel 500, 1000 ppm and  NAA 25, 50 ppm were prepared for seed hardening. PGRs treatments were given to sufficient quantity of seeds of Mung bean cv. GAM-5 for hardening, seeds were soaked in above prepared various solutions of double the volume of seed for three hours. This will ensure that seeds remained immersed in the solution, so as to avoid precocious germination during the treatment period. Hardening was given in flasks under room temperature. The seeds were then removed from respective solutions and kept overnight in shade for drying to attain the seeds to its original moisture level. The seeds were ready for sowing in field on next day.
       
The different solutions with their different concentrations were used in this experiment for foliar spraying at field level. The spraying solutions of CaCl2 1%, Cycocel 500, 1000 ppm and NAA 25, 50 ppm were prepared. The spraying was carried out as per treatments during the morning time or before noon at 30 days after sowing (DAS) in respective gross plot of each replication using knapsack sprayer.
 
Morpho-physiological and growth parameters
 
Plant height, number of branches per plant, number of nodes per plant, days to 50% flowering, days to maturity were recorded by non destructive method from each replication and treatment and the average values were calculated.
 
Leaf area (cm2 plant-1)
 
Leaf area per plant was taken at harvest with the help of Leaf area meter (BIOVIS Company Model- 3100) at Regional Research Station, AAU, Anand. Five plants were randomly selected from all replications and clipped and recorded.
 
Estimation of total leaf chlorophyll content
 
Total chlorophyll content was estimated by the SPAD (soil plant analytical development) meter in which randomly three leaves were selected from lower, middle and upper portion at harvest. Before the measurement, instrument is calibrated - transmission is measured with no leaf inside. Thus, when a leaf is clamped by the meter, a certain portion of red light is absorbed and the meter can calculate a relative value (in SPAD), showing how green the leaf is. Basically, SPAD value correlates with actual chlorophyll content in the leaf, but measurements have to be taken at many points of the same leaf to be representative and calibration is to be performed for every plant species or cultivar to know the exact relationship between SPAD values and chlorophyll contents per unit area.
 
Measurement of seed protein content (%)
 
The protein content was determined by Micro-Kjeldhal method (AOAC, 1990). The method consists of heating a substance with sulphuric acid, which decomposes the organic substance by oxidation to liberate the reduced nitrogen as ammonium sulphate. In this step potassium sulphate was added to increase the boiling point of the medium (from 337°C to 373°C). Chemical decomposition of the sample was completed when the initially very dark-coloured medium has become clear and colourless. The solution was then distilled with a small quantity of sodium hydroxide, which converts the ammonium salt to ammonia. The amount of ammonia present and thus the amount of nitrogen present in the sample, was determined by back titration. The end of the condenser was dipped into a solution of boric acid. The ammonia reacts with the acid and the remainder of the acid was then titrated with a sodium carbonate solution by way of a methyl orange pH indicator.
 
Protein (%) = N (%) × 6.25
 
Yield and yield components
 
Tagged plants used for recording morphological observations were harvested at physiological maturity and were used for recording the yield and yield components viz., number of pods per plant, length of pod (cm), number of seeds per pod, 1000 seed weight (Test weight) (g), seed yield per plant (g plant-1), seed yield per hectare (kg ha-1) and harvest index (%).
Plant height
 
Table 1 represents that at harvest, plant height was maximum in the treatment of CaCl2 2% seed hardening + 1% foliar spraying at 30 DAS (T11) (37.00, 40.67 and 38.83 cm) and remained at par with NAA 50 mg/L seed hardening + spraying at 30 DAS (T15) (35.67, 39.33 and 37.50 cm), NAA 25 mg/L seed hardening + spraying at 30 DAS (T14) (34.67, 37.50 and 36.08 cm) and CaCl2 1%  spraying at 30 DAS (T6) (33.67, 37.00 and 35.33 cm). While Cycocel 1000 mg/L spraying at 30 DAS (T8) recorded significantly less plant height (23.67, 26.00 and 24.83 cm) during the year 2016, 2017 and pooled basis, respectively.
 

Table 1: Effect of seed hardening, foliar spraying and their combined effect on plant height, no. of branches, leaf area at harvest.


       
Plant height was increased due to treatments of CaCl2 and NAA, while decreased due to Cycocel at harvest in green gram. The decrease in plant height with cycocel may be attributed to anti-gibberellic activity of Cycocel mainly by blocking certain steps of gibberellin biosynthesis so that gibberellin is not made available for participation in plant growth (Jain, 2013). The mechanism of reduction of plant height by spraying with Cycocel also appears due to reduced cell size and cell wall thickening (Ginzo et al., 1977). Similarly, Dighe et al., (1983) also observed that 500 ppm cycocel significantly reduced the plant height in wheat.
       
It was observed that NAA showed a positive effect on plant height. The application of auxin in plant has a role in the stimulation of RNA and protein synthesis and greater enhancement in photosynthesis rate, increased in cell elongation as well as cell division and cell wall plasticity, which ultimately showed the enhancement in various growth parameters. The above finding were in agreement with the results reported by Ananthi and Mallika (2014) in green gram, Sunil Jadhav (2016) and Kinjal (2017) in black gram, Upadhyay et al., (2016) in soybean, Pothalkar (2007) in pigeon pea, Amarjeet Singh (2014) in turmeric, Sujatha et al., (2017) in chickpea.
 
Number of branches
 
At harvest, the number of branches per plant found statistically non significant during both the years. However, in pooled mean basis the treatment Cycocel 1000 mg/L seed hardening + spraying at 30 DAS (T13) recorded significantly higher (8.67)  than untreated absolute control with minimum number of branches per plant (6.67).
       
The reduction in plant height due to growth retardants is mainly seems to have released the apical dominance and diversion of the plant metabolites from vertical growth to horizontal growth and thereby more number of branches per plant. These results are in conformity with the findings of Sen (1983) in wheat, Sharma et al., (2003) in pigeon pea, Manjunath and Dhanoji (2011) and Sujatha (2014) in chickpea.
 
Leaf area
 
Leaf area per plant gradually decreased towards harvesting stage in green gram. The data reported in Table 1 indicated that the treatment CaCl2 2% seed hardening + 1% spraying at 30 DAS (T11) registered higher values of leaf area (362, 428 and 395 cm2) over all other treatments. It was followed by T13 (355, 418 and 387 cm2), T15 (344, 404 and 374 cm2) and T12 (341, 395 and 368 cm2) during 2016, 2017 and in pooled analysis, respectively. Whereas, the treatment absolute control (T16) recorded significantly lower value of leaf area per plant (223, 264 and 244 cm2, respectively).
       
The increased in leaf area by seed hardening and foliar spraying with CaCl2 and NAA might be due to increase in cell division, cell enlargement as well as induce more extensive and denser network of veins and ribs and there by increased foliar leaf area. These results are conformity with the finding of Thirumalaiswamy and Rao (1977) in pearl millet, Ginzo et al., (1977) and Josana (2015) in chick pea, Shinde and Jadhav (1995) in cowpea, Pothalkar (2007) in pigeon pea, Prakash et al., (2013) in rice and Kinjal (2017) in black gram.
 
Days to 50% flowering
 
The data indicated in Table 2 showed non significant differences due to various treatments in both the years but in pooled basis differences were significant. In pooled analysis, the treatment T11 (36.67) significantly taken minimum days to 50 per cent flowering followed by the treatments T13 (37.00), T15 (37.17), T12 (37.50) and T14 (37.83). While untreated absolute control (T16) recorded significantly maximum number of days (42.00) to 50 per cent flowering. This might be due to early and faster emergence. Flower initiation is an important phonological development stage which determines the plant productivity.
 

Table 2: Effect of seed hardening, foliar spraying and their combined effect on flowering, maturity, biochemical parameters at harvest.


       
It is inferred that, both the phenological stages viz., flower initiation and pod initiation were early due to seed hardening followed by the use of plant growth regulator’s spray. Similar reports have been also made by Garai and Datta (2003) in green gram and Varma et al., (2004) in pigeon pea. The results in present investigation showed that minimum days taken for 50 per cent flowering in combined effect of seed hardening + foliar spraying followed by only foliar spraying and then only seed hardening treatments. In general, combined effect of seed hardening and foliar spraying was more effective in early flowering as compared to their individual effect.
 
Days to maturity
 
The data indicated non significant differences due to various treatments during both the years but in pooled analysis differences were significant. The treatment T11 (73.17) taken minimum days to maturity followed by the treatments T13 (73.50), T15 (73.83), T12 (74.33) and T14 (74.67). The untreated absolute control (T16) (79.00) taken significantly maximum number of days to maturity.
       
Similar observations on advancement of flowering and harvest times were also reported by Pawar et al., (2003) and Narayanareddy and Biradarpatil (2012) in sunflower, Pothalkar (2007) in pigeonpea, Manjunatha (2007) and Sujatha (2014) in chickpea.
 
Chlorophyll content of leaves
 
The chlorophyll content of leaves gradually decreased towards harvesting stage in green gram. The treatment T11 registered significantly highest values of chlorophyll content of leaves-SPAD values (13.32, 14.80 and 14.06) over absolute control (T16) (9.30, 10.47 and 9.88) and remained at par with T13 (13.22, 14.68 and 13.95), T15 (12.85, 14.43 and 13.64), T12 (12.68, 14.32 and 13.50) and T14 (12.20, 14.00 and 13.10) during 2016, 2017 and on pooled basis respectively.
       
From the data it is clear that continuous increase in chlorophyll content was noted up to 60 DAS, thereafter it decreased during both the seasons at senescence stage. The agrochemicals and growth substances such as CaCl2, Cycocel and NAA had a positive effect on cell division and cell elongation leading to enhanced leaf expansion, leaf area and thereby chlorophyll content also. The increase in chlorophyll content due to growth regulators and agrochemical may be attributed to decreased chlorophyll degradation and increased chlorophyll synthesis. These results are in accordance with Jayakumar and Thangaraj (1998), Shinde and Jadhav (1995), Dashora and Jain (1994), Pothalkar (2007) and Kinjal (2017) in groundnut, cowpea, soybean, pigeon pea and blackgram respectively. In the present investigation, the chlorophyll content showed a positive correlation with grain yield indicating its importance in yield determination.
 
Seed protein content (%)
 
The data presented in Table 2 indicated that even though, the statistically non significant differences observed for the seed protein content during both the years but protein content was improved due to the different treatments. The two years pooled data revealed that the seed protein content recorded maximum (25.76%) in the treatment T11 and it remained at par with the treatments T13 (25.34%), T15 (25.13%), T12 (25.05%), T14 (24.88%) and T6 (24.56%) in pooled. While, the untreated absolute control treatment (T16) recorded significantly lowest (22.82%) seed protein content.
       
From the above results, it is clear that there was increase in seed protein content in all the treatments like seed hardening, foliar spraying singly or their combined effect as compared to control. Similarly, the higher seed protein content was reported by Sujatha (2014) in chickpea. Avijit and Misra, (1987) in wheat, Doijode (1975) in garden peas, Bora and Sarma (2005) in pea. Shukla et al., (2018) in chickpea and Kumar et al., (2015) in field bean and Damor and Patel (2018) in mung bean.
 
Total number of pods per plant
 
The data regarding total number of pods per plant influenced due to different treatments during both the years and on pooled basis were recorded and analyzed are recorded in Table 3. The significantly highest total number of pods per plant (33.73, 35.67 and 34.70) were recorded by the treatment T11, while lowest observed in absolute control (22.00, 23.33 and 22.67) and remained at par with the treatments T13 (32.80, 34.93 and 33.87), T15 (32.13, 34.47 and 33.30), T12 (31.60, 33.67 and 32.63) and T14 (31.00, 33.07 and 32.03) during 2016, 2017 and in pooled, respectively.
 

Table 3: Effect of seed hardening, foliar spraying and their combined effect on pod characters at harvest.



Table 4: Effect of seed hardening, foliar spraying and their combined effect on seed yield and harvest index at harvest.


 
Pod length
 
The significantly highest pod length (8.45) was recorded in the treatment CaCl2 2% seed hardening + 1% spraying at 30 DAS (T11) in pooled analysis. The treatment T11 was also remained at par with the treatments T13 (8.40), T15 (8.33), T12 (8.03) and T14 (7.98). The treatment untreated absolute control (T16) recorded significantly the lowest pod length (7.10).
 
Number of seeds per pod
 
The treatment T11 recorded significantly higher number of seeds per pod (11.93) on pooled basis and remained at par with the treatments T13 (11.83), T15 (11.57), T12 (11.43), T14 (11.33), T6 (11.08) and T8 (11.00). While, the treatment absolute control (T16) recorded significantly lowest (9.73) number of seeds per pod.
 
Test weight (g)
 
The treatment CaCl2 2% seed hardening + 1% spraying at 30 DAS (T11) recorded significantly maximum (53.80 g) thousand seeds weight in pooled analysis and remained at par with the treatments viz., T13, T15, T12, T14, T6, T8, T10, T7 and T9. Whereas, the treatment of absolute control (T16) recorded significantly the minimum (49.27 g) thousand seed weight.
 
Seed yield per plant (g plant-1)
 
The treatment CaCl2 2% seed hardening + 1% spraying at 30 DAS (T11) recorded significantly higher values of seed yield per plant (13.07, 14.20 and 13.63 g) and remained at par with T13 (12.60, 13.93 and 13.27 g), T15 (12.47, 13.60 and 13.03 g) and T12 (12.13, 13.27 and 12.70 g) during 2016, 2017 and on pooled basis, respectively. Whereas, the treatment of absolute control (T16) recorded significantly the lowest seed yield per plant (8.07, 9.40 and 8.73 g, respectively).
 
Seed yield (kg ha-1)
 
The significantly highest seed yield per hectare (949, 1006 and 978 kg ha-1) was recorded by the treatment T11 while significantly the lowest was observed in the absolute control (639, 679 and 659 kg ha-1) during 2016, 2017 and in pooled analysis, respectively and remained at par with the treatments T13 (922, 964 and 943 kg ha-1) and T15 (893, 917 and 905 kg ha-1).
 
Harvest Index (%)
 
The treatment CaCl2 2% seed hardening + 1% spraying at 30 DAS (T11) recorded significantly highest harvest index (30.15%) in pooled analysis and remained at par with the treatments T13 (30.01%), T15 (29.84%), T12 (29.49%), T14 (29.27%), T6 (28.80%), T8 (28.66), T10 (28.35%) and T7 (28.34%). While, the treatment of absolute control (T16) recorded significantly the lowest (26.84%) harvest index.
       
Grain yield is the manifestation of morphological, physiological, biochemical, biophysical and growth parameters. Improvement in yield according to Humphries (1979) could happen in two ways i.e., by adopting the existing varieties to grow better in their environment or by altering the relative proportion of different plant parts so as to increase the yield of economically important parts. The influence of plant growth regulators and seed hardening chemicals significantly increased the seed yield.
       
In the present investigation, it is observed that the number of pods per plant, pod length, seed yield, number of seeds per pod, 1000-seed weight increased due to seed hardening or foliar spraying alone and/or in combination of both by agrochemical CaCl2 and growth regulators NAA and Cycocel.
       
The present study also revealed that increase in seed yield was significantly higher in seed hardening + spraying with CaCl2 (2% seed hardening and 1% spraying) followed by Cycocel 1000 mg/L and NAA 50 mg/L. This could probably be due to beneficial effects of agrochemical and plant growth regulator treatments which help in enhancement of photosynthesis and nitrogen metabolism which are the major physiological process influencing plant growth and development. The treatments of CaCl2 was significantly superior as compared to other treatments in enhancing the plant height, days to flowering, chlorophyll content, number of seeds per plant, 1000-seed weight and thereby seed yield. The increase in the higher yield may be due to better carbon assimilation, better accumulation of carbohydrates and reduced respiration in plants. These results are in agreement with the findings of Singh and Rajodia (1989) in soybean, Singh and Dohare (1964), Das and Prusty (1982) and Pothalkar (2007) in pigeon pea. The present studies also indicated that growth regulators were very effective in increasing yield and yield attributes as compared to control.
       
The present study also indicated that seed hardening + spraying, only foliar spraying and only seed hardening treatments with CaCl2, CCC and NAA significantly increased the number of pods per plant, seed yield per plant, 1000-seed weight and harvest index, which are most important yield determining components in green gram. The increase in seed yield with respect to above treatments was probably due to maximum water absorbing capacity of seeds, more intense photosynthetic activity and more tissue hydration and thereby enabling the plant to resist soil moisture stress more efficiently. This is in conformity with the findings of Mehrotra et al., 1970 in okra, Arjunan and Srinivasan, 1989 in groundnut, Masood Ali (1985) in chickpea, Sen and Misra (1987) in wheat, Patil (1987) in sorghum, Shinde et al., (1991) in cowpea, Singh et al., (1991) in chickpea, Bora and Sarma (2005) in pea, Shinde and Jadhav (1995) in pigeonpea, Amaregouda et al., (1994) in wheat, Jirali (2001) in turmeric and Prabhu (2000) in black gram.
Based on the above results, it is inferred that the yield potential in green gram can be improved by using plant growth regulators and agrochemical. The results also indicated a significant improvement in morpho-physiological traits, growth parameters, biochemical constituents and thereby yield due to the application of PGR’s and agrochemical. Among the different treatments, seed hardening with 2% CaCl2 + 1% foliar spraying at 30 DAS  treatment significantly improved the most of morpho-physiological parameters and thereby yield in green gram  followed by  the seed hardening treatments of Cycocel 1000 mg/L + foliar spraying at 30 DAS and seed hardening with NAA 50 mg/L + foliar spraying at 30 DAS.

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