Environmental Processes & Engineering Efficacy of Algae as Carrier for Efficient Production of Biofertilizer Muhammad Usman Saleem, Rashid Waqas*, Muhammad Arshad, Asif Ali, Zulfiqar Ahmad, Muhammad Imran Institute of Soil and Environmental Sciences, University of Agriculture Faisalabad, Pakistan *Corresponding Author:
[email protected], Tel. +92-41-9201091, Fax. +92-41-9201221 Abstract
In the recent past, world’s attention towards biofertilizers has been tremendously increased which has exerted extra pressure on biofertilizers to perform better. Currently biofertilizer use efficiency is heavily dependent on carrier material for microbes mainly due to the problem of shelf life, consistency and effectiveness. The aim of present study was to evaluate the impact of algae as carrier material to carry plant growth promoting rhizobacteria (PGPR) for efficient growth and yield of mungbean. For this purpose, pre-isolated and pre-characterized strain of PGPR was loaded onto algae by standard method of preparation. PGPR was also inoculated directly on seed in one treatment in addition to the un-inoculated control for comparison. Results showed that inoculation of PGPR on seed has positive impact on all the growth, physiological and yield parameters of mungbean as compared to uninoculated control while this impact was further strengthened when PGPR inoculated on algae used as carrier material. It was concluded that algae could serve as a better carrier material for PGPR and help in production of efficient biofertilizer. Keywords: Algae, Carrier Material, PGPR, Mungbean
1. Introduction Mungbean (Green gram) is one of the most important pulse crops of Pakistan. In 2012, it was grown on an area of 0.140 million hectares and its total production was 0.093 million tons [1], which is very low as compared to the demand of rapidly increasing population. This low production is caused by number of factors which includes low fertilizer use efficiency (FUE), insect pest and diseases, poor seed quality, less area of production and price variability [2]. Among these, FUE is the major contributor of this decline in yield. This low FUE is being tackled by the use of chemical fertilizer in Pakistan. But there are number of problems with the use of chemical fertilizer such as the increasing market prices, unavailability in market at required time, stagnant yields and environmental concerns [3]. This situation demands the alternate approaches to tackle down the problem. In this scenario biofertilizer have emerged to deal the food security issue in combination with chemical fertilizers. In some cases the requirement of chemical fertilizers is reduced up to 50% from the original requirement of the crop by the use of
© Communication for Sciences & Social Sciences
biofertilizers [4]. Combined use of chemical fertilizer and biofertilizer increased the biochemical components, growth and yield of various crops [5]. Biofertilizers are usually the carrier based inoculations containing beneficial microbes. Carrier materials allow the easy handling, increases effectiveness and long storage of biofertilizers with increase in shelf life and support the inocula in its survival [6]. A variety of materials can be used but the need of hour is to find out the most suitable carrier which fulfills all the above stated properties. Among the long list of carrier materials, use of algae offers a great potential towards efficient biofertilizer production because of its autotrophic nature, rapid growth rate, low nutrient requirement and ability to grow in most of the environments. The algae could serve as a better option to carry the microbes without affecting the properties of that microbe. Hence, an alga (Cladophora sp.) was first choice as carrier material in present study. 2. Materials and Methods 2.1. Preparation of Inocula
Environ. Process. Eng., 1-2 (2012), 16–19| 16
For inocula preparation, pre-isolated and pre-characterized strain of PGPR (Bacillus thuringiensis) was obtained from the Environmental Sciences Laboratory of Institute of Soil and Environmental sciences, University of Agriculture Faisalabad. National Botanical Research Institute Phosphate (NBRIP) broth media was used for the inoculum preparation [7]. Inoculation was done in sterilized broth and incubated at 28+1°C & 100 rpm for 72 hours in the shaking incubator. Media was allowed to attain the required population of bacteria (0.6-0.7 optical density). 2.2. Carrier material Alga (Cladophora sp.) was used as carrier material for PGPR inoculation in order to make biofertilizer. These algal carriers were obtained from the Environmental Sciences Laboratory of Institute of Soil and Environmental sciences, University of Agriculture Faisalabad. Carrier material was prepared by grinding, sieving and sterilization before the inoculation. The application of un-inoculated carrier material was also kept for comparison in addition to un-inoculated NPK control.
2.3. PGPR inoculation on seed For inoculation of 100g seeds of mungbean, the seeds were coated with slurry which was prepared by mixing the 10ml sugar solution, 10ml inocula and 50g clay material as adhesive. Inoculated seeds were kept overnight before sowing in the pots.
2.4. Pot trail Pot trail was conducted using mongbean as a test crop in the wire house of Institute of Soil and Environmental Sciences, University of Agriculture Faisalabad, Pakistan to evaluate the impact of self-prepared biofertilizers using algae as carrier material for inocula. Pots were filled with 10 Kg soil belongs to sandy clay loam with ECe (dS m-1) 2.2, pHs 7.6 and Saturation Percentage (%) 31.5. Chemical fertilizer was applied to all the treatments according to the recommended dose for mungbean. The biofertilizers were applied in the soil except from the seed inoculated. In soil application carriers were applied at the rate of 2 g/pot and inocula 1ml/pot. Experiment was conducted in Completely Randomized Design (CRD) and each treatment was triply replicated.
T1 = T2 = T3 = T4 =
NPK (un-inoculated) NPK + Seed Inoculation NPK + algae (un-inoculated) NPK + Inoculated algae
3. Results and Discussion Table 1 illustrates the effect of PGPR when applied to algal carrier on growth and yield of mungbean. Data recorded at crop maturity regarding growth and yield predominantly reflected very promising and consistent positive impact of inoculation with selected PGPR strains either applied on seed or algal carrier, however the latter being the more efficient. Seed inoculation with PGPR enhanced the grain yield (g/pot) of mungbean by 12% over uninoculated NPK control (Table 1). Similar results were found in case of 100 grain weight, plant dry weight and plant height which were increased by 15, 40 and 21% respectively over uninoculated NPK control. In the same way Fig. 1 and Fig. 2 reflects that seed inoculation enhanced the root length, root weight (g/pot) and No. of nodules per plant by 21, 17 and 12% respectively over uninoculated NPK control. While, Fig. 3 explains the positive impact of seed inoculation which enhanced the photosynthetic rate and evapotranspiration rate by 121 and 45% respectively over the uninoculated NPK control. These results are well supported with work of scientists [8, 9, 10] where efficacy of inoculation on seed has been reported on wheat, sorghum, maize and legumes where yield has been increased upto the 25%. Inoculation with PGPR on algal carrier enhanced the grain yield (g/pot) of mungbean by 15 and 21% over uninoculated algal carrier treatment and uninoculated NPK control respectively (Table 1). Similar results were found in case of 100 grain weight, plant dry weight and plant height which were increased by 32, 35 and 25% respectively over un-inoculated algal carrier treatment and 36, 52 and 32% respectively over uninoculated NPK control. Similarly, inoculation with PGPR on algal
Table 1: Impact of PGPR inoculation on algal carrier for the growth and yield of mungbean Plant Height (cm)
Grain yield (g pot-1)
Plant Dry weight (g pot-1)
100 Grain Weight (g)
Control
35.1c
1.2b
2.0b
4.1c
Seed Inoculation
42.7b
1.5a
2.8a
4.7b
Algae (un-inoculated)
37.0c
1.4ab
2.3b
4.2c
Inoculated Algae
46.5a
1.6a
3.0a
5.6a
2.5. Treatment plan © Communication for Sciences & Social Sciences
Environ. Process. Eng., 1-2 (2012), 16–19| 17
carrier enhanced the root length, root weight (g/pot) and no. of nodules per plant by 24, 22 and 10% respectively over un-inoculated algal carrier treatment and 39, 37 and 18% respectively over uninoculated NPK control (Fig. 1 and Fig. 2). In the same way, Fig. 3 illustrates the positive impact of PGPR inoculation on algal carrier which enhanced the photosynthetic rate and evapotranspiration rate 100 and 50% respectively over un-inoculated algal carrier treatment and by 160 and 65% respectively over the uninoculated NPK. These results are also in line with finding that used algae for the promotive effect of biofertilizers [12].
4. Conclusion The results clearly reveal that growth and yield of mungbean could be improved by using the specific PGPR inoculation. However, more growth and yield production in addition of other physiological processes was recorded in response to PGPR inoculation on algal carrier. So, it can be concluded that algae could serve as better carrier material for the efficient biofertilizer production. Acknowledgement The Environmental Sciences Laboratory of Institute of Soil and Environmental sciences, University of Agriculture Faisalabad, Pakistan has provided substantial support in present research work by providing the bacterial (Bacillus thuringiensis) and algal (Cladophora sp.) strains.
References [1]
[2] Fig 1: Impact of PGPR inoculation on algal carrier for the No. of nodules per plant of mungbean [3]
[4]
[5]
Fig 2: Impact of PGPR inoculation onto algal carrier on the root weight and root length of mungbean
[6]
[7]
[8]
[9]
Fig 3: Impact of PGPR inoculation onto algal carrier on the photosynthetic rate and evapotranspiration rate of mungbean © Communication for Sciences & Social Sciences
Government of Pakistan, Economic Survey of Pakistan, Ministry of Finance, Islamabad, Pakistan (2012). M.A.B Mia, Z.H. Shamsuddin, Z. Wahab, M. Marziah, Effect of plant growth promoting rhizobacteria (PGPR) inoculation on growth and nitrogen incorporation of tissue-cultured Musa plantlets under nitrogen-free hydroponics condition, Aust. J. Crop Sci., 4, 85-90 (2010). J. Lewandroski, J. Tobey, Z. Cook, The interface between agricultural assistance and the environment: chemical fertilizer consumption and area expansion, Land Economics., 73, 404-427 (1997). C.L. Wang, Application of arbuscular mycorrhizal fungi on the growth and production of several horticultural seedlings, 27th Int. Hort. Cong & Exhib. Seoul, Korea, (2006). B.R. Chandrasekar, G. Ambrose, N. Jayabalan, Influence of biofertilizers and nitrogen source level on the growth and yield of Echinochloa frumentacea (Roxb.) Link., J. Agric. Tech., 1, 223-234 (2005). H. Muraleedharan, S. Seshadri, K. Parumal, Booklet on biofertilizer. Shri AMM Murugappa Chettiar Research Centre Taramani, Chennai 600113, p 12-13 (2010). C.S. Nautiyal, An efficient microbiological growth medium for screening phosphate solubilizing microorganisms, FEMS Microbiol. Letters, 170, 265270 (1999). M. Arshad, B. Shaharoona, A. Khalid, R. Waqas. Improving water use efficiency of cereals through ACC deaminase biotechnology, 1st Annual Technical Report, Endowment Fund Secretariat, University of Agriculture Faisalabad, Pakistan, (2009). M. Arshad, B. Shaharoona, A. Khalid, R. Waqas. Improving water use efficiency of cereals through ACC deaminase biotechnology, 2nd Annual Technical Report, Endowment Fund Secretariat, University of Agriculture Faisalabad, Pakistan, (2010). Environ. Process. Eng., 1-2 (2012), 16–19| 18
[10]
[11]
[12]
R. Deaker, R.J. Roughly, I.R. Kennedy, Legume seed inoculation-A review, Nitr. Fix. Aus. Sys., 36, 12751285 (2006). E. Ivanova, E. Teunou, D. Poncelet, Alginate based macrocapsules as inoculants carriers for production of nitrogen biofertilizers, Proced. Balk. Sci. Conf. Biol., 90- 108 (2005). S.A. Haroun, M.H. Hussein, The promotive effect of algal biofertilizers on growth, protein pattern and metabolic activities of Lupinus termis plants grown in siliceous soils, Asian J. Plant Sci., 2, 944-951 (2003).
© Communication for Sciences & Social Sciences
Environ. Process. Eng., 1-2 (2012), 16–19| 19
