INTEGRATED EFFECT OF ALGAL BIOCHAR AND PGPR GROWTH AND PHYSIOLOGY

OF PURSLANE (PORTULACA OLERACEA L.) UNDER DROUGHT

ABSTRACT

Biochar is a black carbon formed through the pyrolysis of organic materials such as

plants and organic wastes. It is mainly used in agriculture to improve the production of various

crops. Plant growth-promoting rhizobacteria (PGPR) are widely used due to their role in

improving growth and yield of crops. Drought is one of the main problems of soils under

semiarid and arid region and a major threat to production of valuable crops. In order to mitigate

this problem, it would be imperative to evaluate the combine effect of algal biochar and PGPR

on growth of purslane under drought stress. Based on this hypothesis, a pot experiment will

be conducted with four different combinations of algal biochar and PGPR treatments under

following level of water; one will be 75% and other will be 50% of field capacity and one level

will be normal. Pre and post soil analysis for physiochemical and biological properties will be

conducted. Growth and yield parameters will be recorded after harvesting the crop. Biochar

will be prepared from algae. The strain of PGPR will be used with seed of Purslane. The data

collected will be analyzed for analysis of variance and means will be compared using least

significance (LSD) test. The result will help to formulate approach for the use of bio-inoculants

for improving growth Purslane (Portulaca oleracea L.) crop drought stress.

MATERIALS AND METHODS

BIOCHAR PREPARATION

The dried algal biomass will be placed in muffle furnace at 300 ̊ C for 60 minutes and

biochar will be obtained and then cooled to room temperature.

BIOCHAR APPLICATION
 Biochar will be applied at a rate of 0.4% per kg of soil, i.e 4g per 1000g of soil.

SOIL AND PLANT NUTRIENT ANALYSIS

Soil samples will be collected and will be air dried and sieved through 2mm stainless

steel sieve and stored in plastic container. The physio-chemical properties of soil samples will

be measured including pH, EC, organic matter and soil bulk density. The nutrients of soil K, P

and NO3-N will also be determined.

PLANT PARAMETER

Physiological parameters like photosynthetic rate, stomatal conductance, chlorophyll

fluorescence, stomatal density and transpiration will be measured by infrared gas analyzer

(IRGA). Data regarding growth parameters like plant height, root length, fresh and dry weight

of root and shoot; will be recorded using standard methods. Total nitrogen and total

phosphorous and total potassium will be determined by using colorimeter.

EXPERIMENTAL DESIGN

Four treatments were applied with 3 level of water according to field capacity. The

experiment will be comprised of four different combinations of algal biochar and PGPR, with

each combination at three water levels according to field capacity. One group of plants will be

given a water level 75% water level and other group has 50% of the normal while the remaining

group will have 100% water level (normal).

T 1: Control

T 2: Algal biochar alone

T 3: PGPR alone

T 4: Both algal biochar and PGPR

will be carried out with four replications. The experiment will be completely randomized

design (CRD). The data regarding growth parameters will be collected in response to applied

ratio. The data will be analyzed using statistical software statistix 8.2 and 9.1 and resultant

mean will be compare using least significance difference test.

INOCULA PREPARATION AND SEED INOCULATION

The bacterial inoculum will be produced by taking a loop of PGPR strain to 200 ml of

LB liquid medium in a 500 ml Erlenmeyer flask, incubated for 48 h at 28 ̊ C and 150 rpm. The

optical density of broth will be adjusted to 0.5 calculated λ 535 nm using spectrophotometer

and will obtain a uniform population of bacteria (108-109) colony-forming units (CFU) mL-1 in

the broth at the time of inoculation. For inoculation, the obtained suspension of inoculum will

be mixed with sterilized algal biochar (200 mL kg-1 algal biochar) and will be incubated for 24

h at 28 ̊ C before being used for seed coating (seed to algal biochar ratio 1.25:1 w/w). Purslane

seed dressing will be done with the inoculated algal biochar mixed with 10 % sterilized sugar

(sucrose) solution in 10:1 ratio. In the case of the non-inoculated control, the seeds will be

coated with the sterilized algal biochar treated with sterilized broth and 10 % sterilized sugar

solution.

References

Ahmad, M., A. U. Rajapaksha, J. E. Lim, M. Zhang, N. Bolan, D. Mohan, M. Vithanage, S.

S. Lee and Y. S. Ok. 2014. Biochar as a sorbent for contaminant management in soil

and water: a review. Chemosphere, 99 19-33.

growth, physiology and yield of wheat under salt stress. Agric Water Manage, 158 61-

68.

Aslam, M., M. Zamir, I. Afzal, M. Yaseen, M. Mubeen and A. Shoaib. 2013. Drought stress,

its effect on maize production and development of drought tolerance through

potassium application. Cercetări Agronomice în Moldova, 46 (2): 99-114.

Atkinson, C. J., J. D. Fitzgerald and N. A. Hipps. 2010. Potential mechanisms for achieving

agricultural benefits from biochar application to temperate soils: a review. Plant Soil,

337 (1-2): 1-18.

Ballard, T. 2000. Impacts of forest management on northern forest soils. For Ecol Manage,

133 (1): 37-42.

Barrow, C. 2012. Biochar: potential for countering land degradation and for improving

agriculture. Appl Geogr, 34 21-28.

Bird, M. I., C. M. Wurster, P. H. de Paula Silva, N. A. Paul and R. de Nys. 2012. Algal

biochar: effects and applications. GCB Bioenergy, 4 (1): 61-69.

Cardozo, K. H., T. Guaratini, M. P. Barros, V. R. Falcão, A. P. Tonon, N. P. Lopes, S.

Campos, M. A. Torres, A. O. Souza and P. Colepicolo. 2007. Metabolites from algae

with economical impact. Comparative Biochemistry and Physiology Part C:

Toxicology & Pharmacology, 146 (1): 60-78.

Chan, K., L. Van Zwieten, I. Meszaros, A. Downie and S. Joseph. 2008. Using poultry litter

biochars as soil amendments. Soil Research, 46 (5): 437-444.

Chen, P., D. Haboudane, N. Tremblay, J. Wang, P. Vigneault and B. Li. 2010. New spectral

indicator assessing the efficiency of crop nitrogen treatment in corn and wheat.

Remote Sens Environ, 114 (9): 1987-1997.

soils across a climatic gradient. Journal of Geophysical Research: Biogeosciences

(2005–2012), 113 (G2):

Čuhel, J., M. Šimek, R. J. Laughlin, D. Bru, D. Chèneby, C. J. Watson and L. Philippot.

2010. Insights into the effect of soil pH on N2O and N2 emissions and denitrifier

community size and activity. Appl Environ Microbiol, 76 (6): 1870-1878.

Davies, B. E. 1974. Loss-on-ignition as an estimate of soil organic matter. Soil Sci Soc Am J,

38 (1): 150-151.

DeLuca, T. H., M. D. MacKenzie and M. J. Gundale. 2009. Biochar effects on soil nutrient

transformations. Biochar for environmental management: Science and technology,

251-270.

Demirbas, A. and G. Arin. 2002. An overview of biomass pyrolysis. Energy sources, 24 (5):

471-482.

Demirbas, A. and T. Ozturk. 2005. Anaerobic digestion of agricultural solid residues.

International journal of green energy, 1 (4): 483-494.

Draper, K. and T. Tomlinson. 2012. Poultry litter biochar—a US perspective. Int Biochar

Initiat,

Earl, H. J. and R. F. Davis. 2003. Effect of drought stress on leaf and whole canopy radiation

use efficiency and yield of maize. Agron J, 95 (3): 688-696.

Gaunt, J. and A. Cowie. 2009. Biochar, greenhouse gas accounting and emissions trading.

Biochar for environmental management: Science and technology, 317-340.

Glaser, B., J. Lehmann and W. Zech. 2002. Ameliorating physical and chemical properties of

highly weathered soils in the tropics with charcoal–a review. Biol Fertility Soils, 35

(4): 219-230.
Godfray, H. C. J., J. R. Beddington, I. R. Crute, L. Haddad, D. Lawrence, J. F. Muir, J.

Pretty, S. Robinson, S. M. Thomas and C. Toulmin. 2010. Food security: the

challenge of feeding 9 billion people. Sci, 327 (5967): 812-818.

Guo, Y., H. Tang, G. Li and D. Xie. 2014. Effects of cow dung biochar amendment on

adsorption and leaching of nutrient from an acid yellow soil irrigated with biogas

slurry. Water, Air, Soil Pollut, 225 (1): 1-13.

Hale, S., K. Hanley, J. Lehmann, A. Zimmerman and G. Cornelissen. 2011. Effects of

chemical, biological, and physical aging as well as soil addition on the sorption of

pyrene to activated carbon and biochar. Environ Sci Technol, 45 (24): 10445-10453.

Hiltner, L. 1904. Über neuere Erfahrungen und Probleme auf dem Gebiete der

Bodenbakteriologie unter besonderer Berücksichtigung der Gründüngung und Brache.

Arbeiten der Deutschen Landwirtschaftlichen Gesellschaft, 98 59-78.

Hlavinka, P., M. Trnka, D. Semeradova, M. Dubrovský, Z. Žalud and M. Možný. 2009.

Effect of drought on yield variability of key crops in Czech Republic. Agric For

Meteorol, 149 (3): 431-442.

Initiative, I. B. 2012. Standardized product definition and product testing guidelines for

biochar that is used in soil. IBI biochar standards,

Jeffery, S., T. M. Bezemer, G. Cornelissen, T. W. Kuyper, J. Lehmann, L. Mommer, S. P.

Sohi, T. F. Voorde, D. A. Wardle and J. W. Groenigen. 2015. The way forward in

biochar research: targeting trade‐offs between the potential wins. GCB Bioenergy, 7

(1): 1-13.

John, R. P., G. Anisha, K. M. Nampoothiri and A. Pandey. 2011. Micro and macroalgal

biomass: a renewable source for bioethanol. Bioresour Technol, 102 (1): 186-193.
Joseph, S., C. Peacocke, J. Lehmann and P. Munroe. 2009. Developing a biochar

classification and test methods. Biochar for Environmental Management–Science and

Technology, 107-126.

Kallarackal, J., J. Milburn and D. Baker. 1990. Water Relations of the Banana. III. Effects of

Controlled Water Stress on Water Potential, Transpiration, Photosynthesis and Leaf

Growth. Funct Plant Biol, 17 (1): 79-90.

Kitchen, N., K. Sudduth and S. Drummond. 1999. Soil electrical conductivity as a crop

productivity measure for claypan soils. J Prod Agric, 12 (4): 607-617.

Kumar, N. P. and A. V. Audipudi. 2015. Exploration of a novel plant growth promoting

bacteria Stenotrophomonas maltophilia AVP27 isolated from the chilli rhizosphere

soil. Int j eng resgen sci, 3 (1): 265-276.

Lehmann, J., J. Skjemstad, S. Sohi, J. Carter, M. Barson, P. Falloon, K. Coleman, P.

Woodbury and E. Krull. 2008. Australian climate–carbon cycle feedback reduced by

soil black carbon. Nat Geosci, 1 (12): 832-835.

Madhaiyan, M., B. S. Reddy, R. Anandham, M. Senthilkumar, S. Poonguzhali, S. Sundaram

and T. Sa. 2006. Plant growth–promoting Methylobacterium induces defense

responses in groundnut (Arachis hypogaea L.) compared with rot pathogens. Curr

Microbiol, 53 (4): 270-276.

MICHAEL, I., M. W. CHRISTOPHER and H. PEDRO. 2011. Algal biochar: effects and

applications.

Mishra, A. K. and V. P. Singh. 2010. A review of drought concepts. J HYDROL, 391 (1):

202-216.

Mohan, D., C. U. Pittman and P. H. Steele. 2006. Pyrolysis of wood/biomass for bio-oil: a

critical review. Energy & Fuels, 20 (3): 848-889.

evaluation of chromium remediation from water using low cost bio-char, a green

adsorbent. J Hazard Mater, 188 (1): 319-333.

Mohan, D., A. Sarswat, Y. S. Ok and C. U. Pittman. 2014. Organic and inorganic

contaminants removal from water with biochar, a renewable, low cost and sustainable

adsorbent–a critical review. Bioresour Technol, 160 191-202.

Neori, A., M. Troell, T. Chopin, C. Yarish, A. Critchley and A. H. Buschmann. 2007. The

need for a balanced ecosystem approach to blue revolution aquaculture. Environment:

Science and Policy for Sustainable Development, 49 (3): 36-43.

Nguyen, B. T., J. Lehmann, J. Kinyangi, R. Smernik, S. J. Riha and M. H. Engelhard. 2009.

Long-term black carbon dynamics in cultivated soil. Biogeochemistry, 92 (1-2): 163-

176.

Novak, J. M., W. J. Busscher, D. L. Laird, M. Ahmedna, D. W. Watts and M. A. Niandou.

2009. Impact of biochar amendment on fertility of a southeastern coastal plain soil.

Soil Sci, 174 (2): 105-112.

Packer, M. 2009. Algal capture of carbon dioxide; biomass generation as a tool for

greenhouse gas mitigation with reference to New Zealand energy strategy and policy.

Energy Policy, 37 (9): 3428-3437.

Rani, M. U., A. Arundhath and G. Reddy. 2014. Screening of rhizobacteria containing plant

growth promoting (PGPR) traits in rhizosphere soils and their role in enhancing

growth of pigeon pea. Afr J Biotechnol, 11 (32):

Roberts, K. G., B. A. Gloy, S. Joseph, N. R. Scott and J. Lehmann. 2009. Life cycle

assessment of biochar systems: Estimating the energetic, economic, and climate

change potential. Environ Sci Technol, 44 (2): 827-833.

Sci Med Res, 21 1-30.

Samsuri, A., F. Sadegh-Zadeh and B. Seh-Bardan. 2014. Characterization of biochars

produced from oil palm and rice husks and their adsorption capacities for heavy

metals. Int J Environ Sci Technol, 11 (4): 967-976.

Sayre, R. 2010. Microalgae: the potential for carbon capture. Bioscience, 60 (9): 722-727.

Smith, J. L., H. P. Collins and V. L. Bailey. 2010. The effect of young biochar on soil

respiration. Soil Biol Biochem, 42 (12): 2345-2347.

Sohi, S., E. Krull, E. Lopez-Capel and R. Bol. 2010. A review of biochar and its use and

function in soil. Advances in agronomy, 105 47-82.

Steiner, C. 2008. Biochar Carbon Sequestration. University of Georgia, Biorefining and

Carbon Cycling Program, Athens, GA,

Tilak, K., N. Ranganayaki, K. Pal, R. De, A. Saxena, C. S. Nautiyal, S. Mittal, A. Tripathi

and B. Johri. 2005. Diversity of plant growth and soil health supporting bacteria. Curr

Sci, 89 (1): 136-150.

Van Zwieten, L., S. Kimber, S. Morris, K. Chan, A. Downie, J. Rust, S. Joseph and A.

Cowie. 2010. Effects of biochar from slow pyrolysis of papermill waste on agronomic

performance and soil fertility. Plant Soil, 327 (1-2): 235-246.

Vendrell, P. F. and J. Zupancic. 1990. Determination of soil nitrate by transnitration of

salicylic acid. Communications in Soil Science & Plant Analysis, 21 (13-16): 1705-

1713.

Walkley, A. and I. A. Black. 1934. An examination of the Degtjareff method for determining

soil organic matter, and a proposed modification of the chromic acid titration method.

Soil Sci, 37 (1): 29-38.