Impact of Pranic Agriculture on Germination and Protein Content of Green Gram (Vigna radiata)

Pranic agriculture is an ancient science in which pranic healing technique is applied to the plants to enhance their growth during the developmental stages, where the subtle energy prana is used. Prana is universal, although for most people invisible. The 3 major sources of prana are air prana, solar prana and ground prana. Plants and trees absorb prana from the air, water, sun and ground. Water absorb prana from air, sunlight and ground. All living things including mammals absorb prana from air, water, food and sunlight. Prana can also be referred to as chi, Ki or bioplasmic energy (Sui, 2015). Every living being including plants, trees and human beings are surrounded by an energy field called bioplasmic body or aura (Kafatos et al., 2015). One can feel, interact and use the energy/prana to people and plants to heal. Scientists have rediscovered this bioplasmic body with the help of Kirlian photography (Kirlian,1942). This energy body promotes plants and animals to keep their bodies healthy and alive. Prana is existential, experimental and experiential (Sui, 2015).
       
Pranic agriculture works on the holistic theory of the environment -friendly farming systems, which includes whole invisible subtle energy (Sui, 2015). The application of the pranic healing technique in farming is called Pranic agriculture which is not a substitute rather complementary in nature. Pranic agriculture can be implemented by anybody with basic knowledge of pranic healing. Earlier research articles have been published about the application of pranic agriculture on plants. There was an increase in the seed germination, shoot length, flowering, fruit yield and storage qualities of pranic treated tomatoes (Jois et al., 2016) and pole beans (Yathindra et al., 2017). Studies on storage keeping qualities and physio-chemical composition of cucumber were reported to be significantly higher in the pranic treated group compared to the control group. Additionally, the antioxidant and polyphenol content were also noticed higher in a pranic group (Asna et al., 2016). However, under the present situation of farming, there is a need to adopt green and eco-friendly farming. Hence, implementation of Pranic agriculture treatment is safe, eco-friendly and cost-effective, which helps to enhance the growth and productivity of the crops.
       
Green gram (Vigna radiata) also known as mung bean, mash or moong is a plant species belonging to the Leguminosae family cultivated across Southeast Asia, East Asia and the Indian subcontinent. In India,it is majorly cultivated in Rajasthan, Karnataka, Maharashtra, Orissa and Andhrapradesh (Muthu et al., 2018). It is one of the important summer crops in India with high nutritional value, short growth period, soil fertility restoration by biological nitrogen fixation and it requires less water for crop growth compared to other legumes (Mehandi et al., 2015). Green gram is a good source of carbohydrate (51%), protein (26%), minerals (4%), vitamins (3%) and fat (1%). Protein places the second largest compound in the green gram with high digestibility. Germination is the traditional and non-thermal process that improves the nutritional quality of cereals and pulses. Germinated seeds contain more secondary metabolites and exhibit more biological activities, thus enhancing the nutritional and medicinal qualities of green gram. Due to its high nutritional and low-cost efficiency, these pulses are referred to as poor man’s meat, which helps to overcome the protein malnutrition in developing countries. People usually consume more green gram sprouts due to their rich in protein, dietary fibre and bioactive compounds (Saren et al., 2018). Germination of a green gram enhances antioxidant, anti-inflammatory, antimicrobial, antidiabetic, antihypertensive and antitumor activities along with nutritive benefits (Tang et al., 2014). The objective of this study was to check the effectiveness of pranic energy on germination and protein content of green grams against non-treated green gram commonly referred to as control groups.

Green gram seeds were procured from the local market in Mysore, Karnataka, India. The infested and discoloured seeds were discarded and only the even-sized green gram seeds were selected for this study.
      
The present research was carried out in December 2019 at the World Pranic Healing Foundation, India – Research Centre, Mysore. Randomly seeds were selected and divided into pranic and control groups. Each group having 25 uniform-sized seeds and was placed in a blotting paper of 9cm diameter and kept in the Petri plate. This was done in triplicate. Thus, the whole experiment comprising 6 Petri plates (3 for pranic, 3 for control). Pranic group received pranic healing technique as mentioned by Sui (2018). Healers projects prana initially by praying to supreme God and spiritual teachers to make them a divine channel of healing. Pranic energy was applied to the seeds and water thrice per week, each session lasted for 5min. Seeds were soaked with distilled water for 9 hours and allowed to germinate in the time-lapse apparatus maintained at 25-30°C and kept for 2 days. The seeds germinated percentage was calculated manually for both pranic and control groups for day 1 and 2. Soaked and sprouted fresh weight was measured using the weighing scale, radical length and radical diameter were measured by using the Image j software (Schneider and Rasband 2012). The following parameters were studied during the germination of a green gram as per ISTA standard Rules 2013.
 
Final germination rate (FGR %)
 
Total seed germinated count was taken after 2 days from the sowing date and expressed as a percentage according to the equation of (Al-Mudaris 1998).  
 
Germination index (GI %)
 
Germination Index was determined according to the equation of (Ranalet al., 2009).  
Water absorption rate (WAR %)
 
WAR was calculated by measuring the initial volume of water added for soaking over the final volume of water leftand determined by the following equation (Alvarez et al., 2019).  
 
Sprouted fresh weight of green gram (g)
 
After 2 days of germination, the fresh weight of the sprouted seeds was recorded by electric balance.
 
Radical length (cm) and diameter(cm)
 
Radical length was taken from the area below the hypocotyl to the tip of the root and the radical diameter was taken from one end of the seed to another end and measured by Image J Software.
 
Seedling vigour index
 
Seedling vigour index was calculated as described by Lay and Gowri (2015).
 
  SVI= Mean Seedling Length (cm) ×Germination percentage
 
Protein analysis of sprouted green gram
 
Protein estimation of sprouted green gram was determined by AOAC (2000) method. Green gram sprouts were homogenized using protein extraction buffer comprising sodium Dodecyl sulphate (0.5%), Tris. HCL (pH 8.0), EDTA (0.5mM) and protein inhibitor cocktail. The homogenates were centrifuged to obtain the aqueous phase. 20% TCA/Acetone (1:1 v/v with a final concentration 10% TCA/ Acetone) was added and incubated 5min for protein precipitation. The precipitate was centrifuged, washed with 80% acetone, air-dried and dissolved in 10mM Tris HCl (pH 8.0) buffer.
 
Quantification of proteins
 
The protein content in the control and test samples were determined in duplicates by Lowry’s method (Waterborg, 2009). In this method, the protein sample is allowed to react withFolin–Ciocalteu reagent and copper sulfate to get a blue-colored complex. The colorwas assessed using a colorimeter. The concentration of the unknown sample can be obtained by using Bovine Serum Albumin as a reference standard which depicts the absorbance values of known protein samples.
 
Statistical analysis
 
The data collected were tabulated, analyzed and interpreted using Microsoft excel and statistical analysis was done using paired t-test in SPSS 16.0.

Final Germination rate
 
Germination is the emergence and development of a plant from seed embryo under favourable condition, which is one of the most important processes in the plant's natural life cycles. Results on germination of green gram revealed a significant difference (p<.05) between pranic and control group. The pranic group had a germination of 98.6±1.3% against the control group 92±2.3%. Seeds that received pranic treatment had 7.17% higher rate of germination than the control group (Table 1). These findings are in agreement with those of (Kumar and Jungyun, 2017) where the germination rate of green gram is 91%. Similarly, earlier research on germination of pole beans in the pranic treated group had 90.2% against the control group of 87.1%.Pranic treated pole beans show a 3.1% higher germination rate than the control (Yathindra et al., 2017).


 
Germination index (%)
 
GI emphasizes both the percentage of germination and its speed. A higher GI value denotes a higher percentage and rate of germination. Pranic treated green gram showed a higher germination index of 24.1±0.1 than the control 22.3±0.3 Hence there was a significant difference (p<.05) between the pranic and control group.
 
Water absorption rate (%)
 
Water absorption is the preliminary and necessary factor for germination. When the water gets into seed through a small aperture called micropyle which makes the seeds swell, this swelling of seeds is accompanied by the higher enzyme activity. During 9 hours of soaking, the pranic treated green gram absorbs 21±0.5% of water whereas the control group absorbs only 16.6±0.6% of water and the result was found to be a significant (p<.01). It was observed that the seeds which were given pranic treatment had 26.50% more water absorption than the control group. Similarly, earlier research found that Orgone charged green gram consumed a greater quantity of water of 118.3 ml than the control group of 109.9 ml (DeMeo, 2010). Here orgone is a scientific spiritual concept of esoteric energy or life force.
 
Soaked fresh weight
 
After 9 hours of soaking, the weight of pranic treated green gram had 3.6±.07 g and control had 2.96±.01 g. It was observed that pranic treated green gram gained more weight than the control. Hence there is a significant difference
(p<.05) between the pranic and control group.
 
Sprouted fresh weight
 
After 2 days of germination, the sprouted fresh weight of the pranic treated green gram had 6.4±.15 g and the control had 4.3±.05 g. Thus, the pranic treated group gained more weight than the control with a significant difference (p<.05).
 
Radical length and diameter
 
Mean radical length and mean radical diameter of a pranic treated green gram had 30.4±.10 mm and 8.96±.17 mm respectively, whereas the control group had 21.8±.02 mm and 7.5 ±.01 mm. Pranic group shows 39.45%  higher  radical length and 19.15 % higher radical diameter than the control (Fig 1).It was seen that seeds treated with pranic energy had significantly more (p<.05) radical length and (p<.001) radical diameter, our findings were similar to the Raghu et al., (2014) were radicle length of green gram was found to be 28.6mm.


 
Seedling vigour index
 
Seedling vigour index determines the level of activity and performance of the seed during germination. Seedling vigour test was found to be higher in the pranic 300.81±12.8 when compared to control 199.87±8.40. (Table 1). There was a significant difference (p<0.05) between the pranic and control group.Increase in seedling vigour index is due to the increase in the germination rate and the seedling length.Thus pranic group had the potential level of activity and better performance of the seed during germination.Our findings support earlier research, it was found that pranic treated papaya had 45% more seedling vigour index than the controlgroup (Prasad and Jois, 2020)
 
Lowry’s method for protein estimation
 
Pulses have a high protein content, which is almost double than cereals and root tubers. Protein places the second largest compound after carbohydrates in green gram (Mubarak et al., 2005). The protein content in the pranic treated green gram was found to be higher 48.10±3.4 mg/g when compared to the control group 18.68±3.2 mg/g. Our findings were similar to other researcher results of 23.42 mg/g of protein content in green gram (Widjajaseputra et al., 2019).Hence there was a significant difference (p<.01) between pranic and control groups. Imbibition of water during soaking increases the metabolic activity of resting seeds which will lead to complex metabolic changes resulting in the onset of the sprouting process. Hence the hydrolytic enzymes such as protease and transaminase get activated.This transaminase activity correlates positively with protein synthesis (Dipnaik and Bathere, 2017). As we can see that pranic treated green gram absorbs more water than the control group, this swelling of seeds is accompanied by higher enzyme activity, hence by this protein content in the pranic treated green gram was found to be high.
       
According, to previous research works, qi energy influences the structures of cellular organs such as mitochondria and metabolism of the cells (Kumar Trivedi et al., 2015). In the same way, when the pranic agriculture protocol is applied to the green gram seeds, probably this may increase the energy level in the seeds, thereby resulting in a greater germination percentage and growth of green gram. The reason behind such enhancement in the germination of a green gram could be because of the external pranic energy that can alter the molecular structure of cells, gene expression, nucleotide polymerization and enzyme activity.  It should also be noted that the application of this pranic energy would help in the increased rate of cellular growth and division. It also enhances the activity of ATPase in the bud tip cells during seed germination of the dividing cells this allows more energy supply to the cells during cell growth, division and differentiation (Bai et al., 2000). which probably results in enhanced germination and protein content of pranic treated green gram.

Pranic group green gram had an overall increase in the germination rate, water absorption rate, germination index, soaked weight, sprouted weight, radical length, radical diameter, seedling vigour index and Protein content than the control group. Further studies are in progress to compare the sensory and nutritional qualities of green gram.

Master Choa Kok Sui, the founder of Pranic Healing. Trustees of World Pranic Healing Foundation, India for encouragement and support. MrPapanna N.S. for the commitment to assisting during pranic healing sessions. All those who contributed to the present study directly and indirectly.