Central University of Rajasthan

DISSERTATION PRESENTATION ON

Combined Assessment of Biochar and

Rhizobacteria on the Growth Promotion of
Chickpea (Cicer arietinum L) Plants

Guided By:
Prof. (Dr.) Pradeep Verma
Department of Microbiology
Central University of Rajasthan
Presented By:
Pratyush Kumar Panigrahi
Enrollment No: 2022MSMB013
M.Sc. Microbiology (4th Sem)
 TABLE OF CONTENT
INTRODUCTION 01

OBJECTIVES 02

MATERIALS AND METHODS

03

RESULTS 04

CONCLUSIONS 05

BIBLIOGRAPHY 06
 INTRODUCTION

Stem
• Chickpea (Cicer arietinum L) is a leguminous Leaves
Fruit
plant that comes under the family of Fabaceae
which is rich in nutrition, fibers and protein
cultivated largely in Rajasthan.
• The production of Chickpea can be positively
Rhizosphere
affected by the application of rhizobacteria but microorganisms Root nodules
Fig. Chickpea plant
area like Rajasthan where the annual rain fall
is poor it affect the chickpea production.
• Biochar which can be formed by pyrolysis is rich in Rich in
Balance the carbon
carbon source and have the ability to hold water pH source
and nutrient for a long duration.

• Thus, the combined assessment of biochar and

plant growth promoting rhizobacteria (PGPR) can Water holding Improve soil
capacity quality
promote the plant growth.
 OBJECTIVES

The specific objectives of the present study were to,

 Production of biochar formation forestry waste by drum method.
 Extraction and identification of plant growth promoting rhizobacteria (PGPR) from soil
and root nodules samples.
 Assessment of plant growth parameters after treatment with biochar and plant growth
promoting rhizobacteria.
 MATERIALS AND METHODS
2 3

Soil and root nodule Physico-chemical Streak on Streak on Streak on

sample collection analysis of soil Burk’s media NGNF media CRYEMA media

01 02 03 04 05 06
Lipase test Amylase Protease IAA Ammonia Catalase
test test screening production test test
Secondary screening of isolated bacteria Continued
5 6
7

Forestry waste Biochar formation from Forestry waste Biochar made from Soil
(hard wood) Forestry waste

11 9 8

Root nodules formed after Harvest and measure the

Measure plant physico- Assessment of
21 days physicochemical parameter
chemical parameter of PGPR to the soil of Chickpea plant after 21
Chickpea plant 2nd cycle
days 1st cycle
 RESULTS
Table Secondary screening of isolated bacterial strains
Bacterial Catalase Protease Amylase Lipase NH3 IAA
isolates activity activity activity activity productio production
n
BPBL-R. A + - + - + -
BPBL-R. B + + + + + -
BPBL-R.C + + + - + +
BPBL-R. D + - + + + -
BPBL-R. E + + + - + +
BPBL-R. F + + - - + -
BPBL-R. G + + + - + -
BPBEL-R.H + + + + + +
BPBEL-R. I - - - - + -
BPBEL-A. B + - - + + -
BPBEL-A.C - + + - + -
BPBEL-A. D - - - - + +
BPBEL-A. E + - - - + +
BPBEL-A. I + - + + + +
BPBL-A. J - - - - + -
BPBEL-N. B + - - + + -
BPBEL-N. I + + + + + +
BPBEL-N. D + + - + + - Continued
BPBEL-N. E - + - - + -
 SECONDARY SCREENING
Out of 20 bacteria sample there are 2 bacteria sample that are selected that are
BPBEL-N.I and BPBEL-R.H.

Fig. (a) bacteria spread on CRYEMA plate and the Fig. (a), (b) and (c) the rhizobacteria is streak on
dilution is 10-5, in figure (b) bacteria spread on CRYEMA media for pure culture isolation, (d), (e)
CRYEMA the dilution taken is 10-7, in figure (c) the and (f) the Azotobacter is streak on Burk’s media
bacteria spread on CRYEMA plate with dilution of 10- and Nourish glucose nitrogen free media.
3
, (d), (e) and (f) the same dilution was taken and the
soil sample spread on nutrient agar plate.
Continued
Fig. Secondary screening of bacteria

Continued
 BIOCHAR FORMATION

Fig. Biochar formation by drum method and crushing of

biochar for amendment

Continued
Fig. Ultra microscopic image of biochar made from forestry
waste. This scanning electron microscopic image is taken from Continued
CU Punjab
 ASSESSMENT OF BIOCHAR AND PGPR TO
CHICKPEA (Cicer arietinum L) PLANT.

• After the isolation of bacteria that is BPBEL-R.H and BPBEL-N.I these

bacteria are ready to apply to the plant.

• The experiment was set up that at first 150 gram of soil was taken in a
pot and added 1.5, 2, and 2.5% of biochar to the soil and the plant's
physical and pigmental parameters were measured for about 21 days.

• After 21 days the new plant was planted with the application of isolated
bacteria for 21 days and measure the different physical and pigmental
properties of the plant.

Continued
Fig. (a) seed germination, (b) the plant growth after 1st week, (c) the plant growth in
2nd week and the image (d) the plant growth in 3rd week.

Continued
MEASUREMENT OF PHYSICAL AND PHYSICAL
PROPERTIES OF PLANT SOIL CONTAINING
BIOCHAR
Table Physical parameters of plant planted in soil
containing biochar at different concentration

Avg. Dry weight in Average height Average leaf

Plant sample
gram (g) (cm) count (No.)

Sample-Control 0.06±0.02 14±2 33±18

Sample 1.5% 0.03±0.005 15±2 31±13

Sample-2% 0.07±0.01 16±2 40.5±5.5

Sample-2.5% 0.04±0.01 13.5±1.5 41±9

Continued
Fig. Graphical presentation of average height Fig. Graphical presentation of average dry
and leaf count before application of biochar weight of plant before application of biochar

Continued
 PIGMENTAL PARAMETER OF PLANT AFTER 21
DAYS SOIL CONTAINING BIOCHAR
Table Concentration of chlorophyll-a, chlorophyll-b and total
carotenoids in different plant, planted in pot contain different
biochar concentration.

Plant sample Avg chla Avg chlb Avg total carotenoid

Control 3.97±2.50 2.22±1.05 0.80±0.47

Soil+BC 1.5% 4.83±3.59 2.60±1.39 0.97±0.57

Soil+BC 2% 8.11±8.76 3.89±2.97 1.34±0.90

Soil+BC 2.5% 5.82±4.07 2.93±1.56 0.95±0.66

665.2652.2 Fig. Average Chlorophyll a, Chlorophyll b and
total carotenoid of plant planted in soil
652.4665.2
containing biochar

𝑇𝐶𝑠
µg
ml ( )
=
(1000 × A ( 470 ) −1.63 ×Chl . a −104.96 ×Chl . b)
221
Continued
 MEASURE PHYSICAL PARAMETERS OF PLANT OF SOIL
CONTAINING BIOCHAR AND MICROBIAL INOCULANT

Table Average height of plant sample containing biochar

and plant growth promoting bacteria.

Plant sample Plant-1 Plant-2 Plant-3 Height (cm)

R+A 19 18 22 19.66±1.69

A+BC 2% 21 20 19 20±0.81

A+BC 1.5% 18 18 19 18.33±0.47

A+BC 2.5% 14 16 16 15.33±0.94

R+BC 1.5% 15 8 11 11.33±2.86

R+BC 2% 9 18 12 13±3.74

R+BC 2.5% 19 18 17 18±0.81

Fig. Graphical presentation of average height

of plant soil containing biochar and microbial
inoculant. Continued
 Table Average leaf number of plant sample containing
biochar and plant growth promoting bacteria.
Average
Plant sample Plant-1 Plant-2 Plant-3
leaf count

R+A 53 57 55 55±1.63

A+BC 1.5% 47 45 37 43±4.32

A+BC 2% 79 41 50 56.66±16.21

A+BC 2.5% 42 52 45 46.33±4.18

R+BC 1.5% 40 42 50 44±4.32

R+BC 2% 21 31 33 28.33±5.24

R+BC 2.5% 51 41 35 42.33±6.59 Fig. Graphical presentation of average leaf

number of plant soil containing biochar and
microbial inoculant
Continued
 Table Average dry weight of plant sample containing
biochar and plant growth promoting bacteria.

Plant sample P=1 P=2 P=3 Average dry weight

A+R 0.07 0.075 0.085 0.07±0.006

R+BC1.5% 0.08 0.102 0.102 0.09±0.01

R+BC2% 0.07 0.07 0.07 0.07±0

R+BC2.5% 0.08 0.06 0.06 0.06±0.009

A+BC1.5% 0.106 0.07 0.07 0.08±0.01

A+BC2% 0.101 0.21 0.21 0.17±0.05 Fig. Graphical presentation of average dry
weight of plant soil containing biochar and
microbial inoculant
A+BC2.5% 0.077 0.09 0.09 0.08±0.006

Continued
 MEASURE PIGMENTAL PARAMETERS OF PLANT OF SOIL
CONTAINING BIOCHAR AND MICROBIAL INOCULANT

Table Concentration of chlorophyll-a, chlorophyll-b and total

carotenoids in different plant, planted in pot contain different
biochar concentration.
total
Plant sample avg chl-a avg.chlb
Carotenoids

A+R 9.57±5.17 7.91±5.35 2.83±1.46

R+BC1.5% 6.62±5.17 3.66±4.35 1.14±0.77

R+BC2% 11.28±12.73 6.03±6.66 1.80±2.35

R+BC2.5% 11.59±9.19 5.53±4.57 1.70±1.42

A+BC1.5% 10.65±8.61 5.02±3.69 1.49±1.16

A+BC2% 13.94±8.99 6.45±3.53 2.32±2.65

Fig. Average Chlorophyll a, Chlorophyll b and
total carotenoid of plant planted in soil
A+BC2.5% 11.85±9.01 6.29±3.99 1.66±1.57
containing biochar and microbial inoculant.
 CONCLUSIONS
_____________________________

• Soil obtained from semi arid and poor rainfall area which create drought like
condition which show poor growth of chickpea (Cicer arietinum L).

• Although biochar in a concentration of 2% gives the better plant growth but

when we combined this concentration of biochar with plant growth promoting
rhizobacteria (PGPR) they give a better result.

• Soil contain the Azotobacter and biochar of 2% can achieve height of 20cm,
79 number of average leaf and 0.173g of dry weight, also the pigments like
chlorophyll-a is higher in this plant and the chlorophyll-b and carotenoid
concentration is high in pot contain mixture of Rhizobacteria and
Azotobacter as compare to the control (soil without any amendment) and soil
containing biochar with in 21 days.
 BIBLIOGRAPHY
______________________________
1. Asari, S., Matzén, S., Petersen, M. A., Bejai, S., & Meijer, J. (2016). Multiple effects of Bacillus amyloliquefaciens volatile compounds:
Plant growth promotion and growth inhibition of phytopathogens. FEMS Microbiology Ecology, 92(6), fiw070.
https://doi.org/10.1093/femsec/fiw070.
2. Becerra, S. C., Roy, D. C., Sanchez, C. J., Christy, R. J., & Burmeister, D. M. (2016). An optimized staining technique for the
detection of Gram positive and Gram negative bacteria within tissue. BMC Research Notes, 9(1), 216.
https://doi.org/10.1186/s13104-016-1902-0.
3. Boekema, B. K. H. L., Beselin, A., Breuer, M., Hauer, B., Koster, M., Rosenau, F., Jaeger, K.-E., & Tommassen, J. (2007).
Hexadecane and tween 80 stimulate lipase production in Burkholderia glumae by different mechanisms. applied and environmental
microbiology, 73(12), 3838–3844. https://doi.org/10.1128/AEM.00097-07.
4. Cochard, B., Giroud, B., Crovadore, J., Chablais, R., Arminjon, L., & Lefort, F. (2022). Endophytic PGPR from Tomato Roots:
Isolation, In Vitro Characterization and In Vivo Evaluation of Treated Tomatoes (Solanum lycopersicum L.). Microorganisms, 10(4),
765. https://doi.org/10.3390/microorganisms10040765.
5. Debiagi, P., Gentile, G., Cuoci, A., Frassoldati, A., Ranzi, E., & Faravelli, T. (2018). A predictive model of biochar formation and
characterization. Journal of Analytical and Applied Pyrolysis, 134, 326–335. https://doi.org/10.1016/j.jaap.2018.06.022.
THANK YOU.
FOR YOUR ATTENTION