Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Gangwar M, et al. (2016) Molecular Analysis of Bioeld Treated Eggplant and Watermelon Crops.
Adv Crop Sci Tech 4: 208. doi:10.4172/2329-8863.1000208
Page 2 of 5
Volume 4 • Issue 1 • 1000208
Adv Crop Sci Tech
ISSN: 2329-8863 ACST, an open access journal
Materials and Methods
Eggplant (Solanum melongena) and watermelon (Citrullus lanatus)
were selected for the present study. Both the plants were selected from
inbreed variety for all the experimental parameters. e bioeld treated
plot size for eggplant was 64 × 8 feet, while the control plot size was
47 × 12 feet. e treated plot size for watermelon was 64 × 16 feet,
while the control plot size was 35 × 25 feet. Both the plots were have
same number of plants, and were compared with respect to respective
control. e control plots were le untreated, while the treated plots
of eggplant and watermelon was subjected to Mr. Trivedi’s bioeld
energy treatment. e seeds from each crop were cultivated for analysis.
However, the control plants were given standard cultivation parameters
such as proper irrigation, fertilizers, pesticides and fungicides; while
the treated plots were given only irrigation, without any supportive
measure. DNA ngerprinting of both the plants were performed using
RAPD techniques using Ultrapure Genomic DNA Prep Kit; Cat KT 83
(Bangalore Genei, India) to study the genetic relationship before and
Bioeld treatment strategy
e treated plots were subjected to Mr. Trivedi’s bioeld energy
treatment. Mr. Trivedi provided the unique bioeld treatment through
his energy transmission process to the selected treated plots of both
the crops. e plant samples of treated plots were assessed for the
growth attributes with respect to control. Variability in dierent growth
contributing parameters and genetic relatedness using RAPD of control
and treated crops were compared .
Analysis of growth and related parameters of crops
e seeds of eggplant and watermelon were cultivated under similar
conditions. e vegetative growth of the crops with respect to plant canopy,
the shape of leaves, owering conditions, infection rate, etc. were analyzed
and compared with respect to the plants of control plots .
DNA ngerprinting isolation of plant genomic DNA using
e leaves disc of both plants were harvested aer germination, as it
reached the appropriate stage. e genomic DNA from both plant leaves
was isolated according to the standard cetyl-trimethyl-ammonium
bromide (CTAB) method . Approximately 200 mg of plant tissues
were grinded to a ne paste in approximately 500 μL of CTAB buer.
e mixture (CTAB/plant extract) was transferred to a microcentrifuge
tube, and incubated for about 15 min at 55
C in a recirculating water
bath. Aer incubation, the mixture was centrifuged at 12000 g for 5
min and the supernatant was transferred to a clean microcentrifuge
tube. Aer mixing with chloroform and iso-amyl alcohol followed
by centrifugation the aqueous layers were isolated which contain the
DNA. en, ammonium acetate followed by chilled absolute ethanol
were added, to precipitate the DNA content and stored at -20
RNase treatment was provided to remove any RNA material followed
by washing with DNA free sterile solution. e quantity of genomic
DNA was measured at 260 nm using spectrophotometer .
Random amplied polymorphic DNA (RAPD) analysis
e RAPD analysis was performed on the each treated plot
plants using RAPD primers, which were label as RPL 6A, RPL 13A,
RPL 16A, RPL 18A, and RPL 19A for eggplant, while RPL 2A, RPL
7A, RPL 12A, RPL 14A, RPL 18A, and RPL 23A for watermelon. e
DNA concentration was considered about 25 ng/µL using distilled
deionized water for polymerase chain reaction (PCR) experiment.
e PCR mixture including 2.5 μL each of buer, 4.0 mM each of
dNTP, 2.5 μM each of primer, 5.0 μL (approximately 20 ng) of each
genomic DNA, 2U each of ermus aquaticus (Taq) polymerase, 1.5
μL of MgCl
and 9.5 μL of water in a total of 25 μL with the following
PCR amplication protocol. e PCR cycle condition for eggplant and
watermelon includes initial denaturation at 94
C for 5 min, followed by
40 cycles of annealing at 94
C for 1 min, annealing at 36
C for 1 min,
and extension at 72
C for 2 min, while nal extension was carried out at
C for 10 min. Amplied PCR products (12 µL of each) from control
and treated samples were loaded on to 1.5% agarose gel and resolved
by electrophoresis at 75 volts. Each fragment was estimated using 100
bp ladder (Genei
; Cat # RMBD19S). e watermelon sample was
analyzed with help another ladder of 500 bp ladder (Genei
; Cat #
RMBD13S). e gel was subsequently stained with ethidium bromide
and viewed under UV-light . Photographs were documented
subsequently. e following formula was used for calculation of the
percentage of polymorphism.
Percent polymorphism = A/B × 100
Where, A = number of polymorphic bands in treated plant; and B =
number of polymorphic bands in the control plant.
Results and Discussion
Eect of bioeld treatment on growth contributing
parameters of eggplant
e eggplant crop in control plots showed the survival as less than
60-65%. e growth of eggplants was much less in the control group as
well. e eggplants had a small canopy in control plot. e leaves were
small in size, and their color was light green in eggplants of control plot.
e fruit of eggplant in control plot did not have uniform shape and
most of the fruit was diseased even aer being sprayed with pesticides
On the other hand, the bioeld treated plot for eggplants showed
the leaves were thick, glossy, more green in color and bigger in size.
e canopy of the eggplant in bioeld treated plot was also larger as
compared with the control crop. e budding was early, that suggest
early fruiting as compared with the control. In bioeld treated plots, all
eggplants fruits had uniform shape and the texture as compared with
the control. Further, no disease in the treated plants were observed as
compared with the control (Figure 1).
Research study suggest that both mineral fertilizers and organic
manures have their own roles in soil fertility management, however
none can completely provide all the nutrients and conditions, which may
enhance the growth of eggplant . Bioeld treatment on soil selected
for eggplant crops, showed enhanced growth in the absence of chemical
fertilizers, and suggested the alternate method to improve the crop yield.
Eect of bioeld treatment on growth contributing
parameters of watermelon
In the control plot, the survival rate of the watermelon plants was
less than 60 to 65%. e canopy was small, and the color of the leaves
were pale green in plants of control plot. Many of the plants were
reported as diseased and even the fruits were infected at an early stage,
with small fruit size in plants of control plot.
e bioeld energy treatment on plots with watermelon plants
showed high survival rate i.e., more than 99%. e canopy of watermelon
plants was much larger in treated plots than in the control plot plants.