The links below provide some information on EM Technology® for Bioremediation. For a full database of research papers on Effective Microorganisms®, please visit EMRO Japan's website.
Effect of Different Fertilizers and Effective Microorganisms on Growth, Yield and Quality of Maize
SHAMSHAD HUSSAIN SHAH, M. FARRUKH SALEEM AND M. SHAHID
Department of Agronomy, University of Agriculture, Faisalabad–38040, Pakistan
The studies were carried out to determine the effect of different combinations of N, P, FYM and effective microorganisms (EM) on growth, yield and quality of maize (Zea mays L.) during autumn 1998. Results revealed that highest grain yield of 4.72 t ha-1 was obtained with the application of 150 kg N + 75 kg P2O5 + 30 l EM ha-1. The increase in yield was attributed to increased leaf area, more number of grains per cob and higher weight per 1000 grains. The protein content (10.03%) were however, higher with the application of 75 kg N + 37.5 kg P2O5 + 60 l EM ha-1.
Key Words: Fertilizer; Organic; Inorganic; EM; Maize yield
Mineral fertilizers are important and quickest way of nutrient supply to soil and play an important role in activating various enzymes (Tisdale et al., 1990) However, in addition to other constraints, their high cost and short supply at the time of need deter the farmers from using recommended doses (FAO, 1978). This necessitates to explore alternative potential sources of plant nutrients (organic) with the minimum use of mineral fertilizers. Organic matter is of great importance for the maintenance of soil structure, soil bioactivity, soil exchange capacity and water holding capacity (NFDC, 1998). Organic materials like FYM are used for increasing crop production but pure organic farming can never meet the increasing demand for nutrient supply, as sufficient quantities of organic materials are not available. Another way of supplying nutrients to soil is through biological inoculum but it also needs large amount of organic matter and alone cannot favour the plant nutrient supply to soil eco-system (Hussain et al., 1999). So, one of the alternative of nutrient supply is the integration of Effective Microorganisms (EM) inoculum and organic/inorganic materials. The present studies were conducted to develop a production system for the efficient utilization of mineral and organic nutrient resources by amending with EM.
MATERIALS AND METHODS
The studies were carried out at the Agronomic Research Area, University of Agriculture, Faisalabad. The experiment comprised of nine treatments i.e. control 12.5 tonnes FYM + 60 l EM; 75 kg N + 60 l EM+; 37.5 kg P2O5 +60lEM+;75kg N+37.5kgP2O5 +60lEM; 25tonnes FYM+30lEM; 150kg N+30lEM; 75kg P2O5 +30l EM;150kgN+75kg P2O5 +30l EMha-1.The crop was sown on a well prepared seedbed in the first week of August, 1998 using a seed rate of 30 kg ha-1. The whole of P, FYM and 1/3rd N were applied at sowing and EM was added with each of the nine irrigations. Remaining N was applied in two splits i.e. when the crop height was 45 cm and at tasseling stage. The experiment was laid out in a randomized complete block design (RCBD) with three repeats and a net plot size of 3m x 5m was used. All the agronomic operations except those under study were kept normal and uniform for all the treatments. The crop was harvested on November 12, 1998. Data on various growth and yield components were collected using standard procedures and analyzed statistically by using Fisher’s analysis of variance technique (Steel & Torrie, 1984). Least significant difference (LSD) test at 0.05 probability was employed to compare the means.
RESULTS AND DISCUSSION
Data on final plant height (Table I) indicate that none of the combinations of fertilizer and EM could reach a level of significance. However, maximum plant height of 244.50 cm was recorded in T5 where crop was treated with 25 t ha-1 FYM + 30 1 ha-1 EM and it was the lowest (203.1 cm) in the control (T0). These results are in agreement with those of Ghafoor and Akhtar (1991) who reported that increased fertilization had non-significant effect on plant height of maize cultivar. Maximum number of grains per cob 572.40 was obtained in T8 where N, P and EM were applied @ 150, 75 kg ha-1 and 30 l ha-1, respectively. Treatment T2 did not differ significantly from either T3 or T1, which produced 344.50, 334.00 and 308.90 grains per cob, respectively. The lowest number of grains per cob (270.90) was produced in the control, treated with no fertilizer. The results indicate that the number of grains per cob increased with increase in the doses of fertilizer. These findings are in agreement with those of Short et al. (1982) and Choudhary (1997). It is evident from the data presented in Table I that maximum 1000-grain weight of 234.30 g was obtained in treatment T8 where nitrogen was applied @ 150 kg ha-1 but SHAH et al. / Int. J. Agri. Biol., Vol. 3, No. 4, 2001
It did not differ significantly from that recorded in treatment T4 receiving 75 kg N, 37.5 kg P2O5 and 60 l EM ha-1. Non-significant difference was found between treatments T3 and T1 but they were significantly different from that of T2 and T5, which were at par with each other. The lowest weight per 1000-grains of 195.60 g was recorded in control treatment. Weight per 1000-grains increased with the increase in N level in addition to P and EM. These findings are in conformity with those of Ahmad (1989) and Choudhary (1997). It also transpires from Table I that fertilizer combinations along with EM had a highly significant effect on the grain yield. Grain yield increased with increase in N levels and it was maximum (4.72 t ha-1) when crop was treated with the highest dose of nitrogen (150kg ha-1) in addition to P and EM @ 75kg ha-1 and 30 1 ha-1 , respectively. It was followed by T4 (75 kg N ha-1 ), which produced 4.16 t of grains ha-1. Grain yield recorded in control treatment was the lowest (1.65 t ha-1) of all other treatments. The higher grain yield at the highest dose of N (150 kg ha-1) was attributed to greater leaf area, more number of grains per cob and higher weight per 1000-grains. These results are in line with those obtained by Panchaban (1991), Rashid (1993) and Shahzad et al. (1996). Protein percentage in grains was affected significantly by different fertilizer combinations with EM. All the fertilizer combinations significantly enhanced the protein content in grains over the control. The highest protein content (10.03%) in grains was recorded with the application of 75 kgN ha-1 in addition to P and EM @ 75kgha-1 and 30 l ha- 1, respectively which was statistically at par with T8 (150 kg N + 75 kg P2O5 + 30 l EM ha-1). The protein content in treatments T5 and T7 were statistically at par and were higher than that recorded in T6, T1 and T2 treatments which were statistically the same with one another. The lowest protein content recorded in the control was 9.23%. Similar results were reported by Hera et al. (1982), Feng et al. (1993) and Iqbal (1997).
The results indicated that maize cultivar Composite-17 should be fertilized @ 150 kg N ha-1 and 75 kg P2O5 ha-1 along with 30 l EM ha-1 to obtain maximum yield and highest grain protein content under ecological conditions of Faisalabad.
Choudhary, M.A., 1997. Effect of various doses of nitrogen at a constant level of phosphorus and potash on the growth, yield and quality of two maize (Zea mays L.) cultivars. M.Sc. (Hons.) Agri. Thesis, Deptt. Agron. Univ. Agric., Faisalabad.
FAO, 1978. Organic Recycling in Asia, Soil Bulletin No. 36, p. iv. FAO, Rome.
Feng, Z., A.F. Macheazie and D.L. Smith, 1993. Corn yield and shifts among corn quality constituents following application of different nitrogen fertilizer source at several times during corn development. J. PI. Nutr., 16: 1317–37 (Maize Absts., 10(4): 1991; 1994).
Ghafoor, A. and S. Akhtar, 1991. Response of spring maize cv. Agatti–72 to nitrogen fertilization under saline sodic soil culture. Pakistan J. Agric. Sci., 23: 374–8.
Hera, C., A. Indreceanu, S. Popescu, S. Stan and I. Vines, 1982. Influence of fertilization on maize quality. Analde institutului de cercetari pentru areal Si plante technice fundulea. Romania, 50: 255–63 (Field Crop Absts., 37(6): 3854; 1984).
Hussain, T., T. Javaid, J.F. Parr, G. Jilani and M.A. Haq, 1999. Rice and wheat Production in Pakistan with Effective Microorganisms. A. J. Alt. Agri., 14: 30–6.
Iqbal, M., 1997. Growth and yield response of hybrid maize to nitrogen application. M.Sc. (Hons.) Agri. Thesis, Deptt. Agron., Univ. Agri., Faisalabad.
NFDC, 1998. Integrated Plant Nutrition System (IPNS). NFDC technical report 3/98, p 30. Planning and Development Division. National Fertilizer Development Centre, Islamabad, Pakistan.
Panchaban, S., 1991. Effect of EM on growth and yield of corn. Proc. 1st Intl. Conf. on Kyusu Nature Farming, pp: 132–9. Oct. 17–21, 1989, Khon Kaen, Thailand.
Shahzad, M.A., M. Musa, G.A. Chaudhry, M. Nasim and M.A. Gondal, 1996. Response of maize of NPK application under barani conditions. Pakistan J. Soil Sci., 12: 74–7.
Short, K.E., M.S. Islam, A.A., Pahlwan and S.K. Roy, 1982. Yield response of maize to different rates of nitrogen and sulfur during the Kharif season in Bangladesh. Bangladesh J. Agric., 7: 40–7 (Field Crop Absts, 37(9): 2185; 1984).
Steel, R.G.D. and J.H. Torrie, 1984. Principles and Practices of Statistics, 2nd Ed., pp: 107–9. McGraw Hill Co. Inc. New York.
Tisdale, S.L., W.L. Nelson and J.D. Beaton, 1990. Soil Fertility and Fertilizer: Elements Required in Plant Nutrition, 4th Ed., pp: 52–92. Maxwell Macmillan Pub., Singapore.
(Received 14 July 2001; Accepted 20 August 2001)
INTERNATIONAL JOURNAL OF AGRICULTURE & BIOLOGY 1560–8530/2001/03–4–378–379 http://www.ijab.org
Growth, nodulation and yield of black gram [Vigna mungo (L.) Hepper] as influenced by biofertilizers and soil amendments
Institute of Mycology and Plant Pathology, University of the Punjab, Quaid-e-Azam Campus Lahore, Pakistan. E-mail: firstname.lastname@example.org.
Accepted 7 August, 2009
EM (effective microorganisms) is a commercial biofertilizer mainly consists of photosynthetic and lactic acid bacteria, yeast and actinomycetes. The present study was undertaken to investigate the effect of EM application and two strains of nitrogen fixing Bradyrhizobium japonicum (TAL- 102 and MN-S) on plant growth, nodulation and yield of black gram [Vigna mungo (L.) Hepper] in different soil amendment systems including unamended soil, farmyard manure (FYM) @ 5 g 100 g-1, Trifolium alexandrinum green manure (GM) @ 4 g 100 g-1 and recommended dose of NPK fertilizers. Nodule number was significantly enhanced by inoculation of either of the two B. japonicum strains in NPK and un-amended soils. A marked increase in nodule biomass was also recorded due to B. japonicum inoculation in these 2 types of soils. Grain yield was significantly increased by 46% due to either of the two B. japonicum strains in NPK amended soil. EM application markedly enhanced nodule number in FYM amended soil. Conversely, EM application in combination with either of the two B. japonicum strains resulted in pronounced reduction both in number and biomass of nodules in NPK fertilizers amendment. EM application significantly enhanced grain yield by 48% in NPK amendment without B. japonicum inoculation.
Key words: Black grams, Bradyrhizobium japonicum, effective microorganisms, nitrogen fixation, soil amendments.
Chemical fertilizers are an indispensable component of today’s agriculture. About 60% of humanity eventually owes its nutritional survival to N fertilizers (Fixon and West, 2002). However, growing concern about the environmental consequences of mineral N use and its future cost perspectives emphasize the need to develop new production technologies that are sustainable both economically and ecologically (Khaliq et al., 2006). Organic materials hold great promise as a source of multiple nutrients and ability to improve soil characteris- tics (Soumare et al., 2003; Moller, 2009). Since the effect of organic nutrients on crop yield is long term and not immediate, thus farmers are reluctant to use organic fertilizers in their cropping system. Use of EM (effective microorganisms) along with organic materials possibly be an effective technique for stimulating release of nutrients from organic sources. EM technology was developed by Dr. Teuro Higa in 1970’s at the University of Ryukyus, Okinawa, Japan. Effective microorganisms culture con- sists of co-existing beneficial microorganisms, the main being the species of photosynthetic bacteria; Rhodopseu- domonas plastris and Rhodobacter sphacrodes; lacto- bacilli such as Lactobacillus plantarum, L. casei and Streptococcus lactis; yeasts (Saccharomyces spp) and Actinomycetes (Strptomyces spp.) which improve crop growth and yield by increasing photosynthesis, producing bioactive substances such as hormones and enzymes, controlling soil diseases and accelerating decomposition of lignin materials in the soil (Higa, 2000; Hussain et al., 2002). When effective micro-organisms cultures are applied to the soil they stimulate the decomposition of organic wastes and residues thereby releasing inorganic nutrients for plant uptake. Majority of the scientists who are engaged in promoting this technology have no doubt that plant growth is just as good or batter and quality of plant products is superior to conventional farming (Bajwa et al., 1999a; Iwaishi, 2000; Xu et al., 2000; Javaid, 2006). However, experiences of some workers revealed that the effect of effective microorganisms on crop yield was usually not evident or even negative particularly in the first test crop (Javaid et al., 2008). It is often difficult to establish the predominance of effective microorganisms cultures in soil with only a single application and during only one season. Certain soil properties and the indigenous soil microbial populations are often constraints to the establishment of these microorganisms (Bajwa et al., 1995; Javaid et al., 1997).
Black gram is a grain legume widely cultivated in Pakistan, India and other Asian countries. It is part of diet for millions of people in these countries and a cheep source of protein with 17 - 34% of protein in seeds (Gour, 1993). An important feature of the mashbean plant is its ability to establish a symbiotic partnership with specific bacteria, setting up the biological N2-fixation process in root nodules by rhizobia that may supply the plant's needs for N (Mahmood and Athar, 2008; Mandal et al., 2009). The present study was carried out to investigate the effect of two Bradyrhizobium japonicum strains; TAL- 102 (soybean isolate) and MN-S (mungbean isolate) on growth, nodulation and yield of mashbean and role of EM in improving the efficacy of these strains in different soil amendment systems.
MATERIALS AND METHODS
Soil used in the experiment was sandy loam in texture having organic matter 0.9%, pH 8.1, EC 4.8 mS cm-1, nitrogen 0.05%, available phosphorus 14 mg.kg-1 and available potassium 210 mg.kg-1. The micronutrients Fe, Cu and Zn were 9.53, 1.71 and 4.42 mg kg-1 of soil, respectively.
This experiment was a continuation of a previous experiment where mungbean [Vigna radiata (L.) Wilczek] was cultivated. Experiment was conducted in earthen pots of 20 cm diameter and 30 cm deep. The pot soil was amended either with farmyard manure (FYM) @ 5 g/100 g, Trifolium alexandrianum green manure (GM) @ 4 g/100 g (on dry weight basis), NPK fertilizers or left unamended. A basal dose of 20 mg kg-1 N as urea, 30 mg kg-1 P2O5 as triple supper phosphate and 30 mg kg-1 K2O as potassium sulphate was supplied to the NPK amended pot soil. Pots were irrigated with tap water of good quality and left for 15 days for decomposition of organic matter. Mungbean was sown in these pots. After harvesting the mungbean crop, the present study was conducted in the same pots.
No more GM and FYM were added for the present study. However, NPK fertilizers were added to the respective pots at the same rate as was for mungbean mentioned above.
Procurement of B. japonicum and EM
Two peat based B. japonicum inocula namely B. japonicum st. TAL- 102 and B. japonicum st. MN-S were obtained from Nuclear Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan. B. japonicum st. TAL- 102 is an exotic strain originally isolated from soybean while B. japonicum st. MN-S is a local strain isolated from mungbean.
EM stock solution in the commercial name of EM Bioaab was obtained from Nature Farming Research and Development Foundation, Faisalabad, Pakistan. The EM contained high popu- lations of lactic acid bacteria at 1 × 1011 cfu ml-1, photosynthetic bacteria at 1 × 106 cfu ml-1 and yeast 1 × 103 cfu ml-1 of suspension (Higa, 2000). The EM stock solution was diluted by adding water in the ratio of 1:1000. Respective pots of EM treatments were irrigated with 500 ml of dilute solution of EM (1:1000) 15 days prior to mung- bean sowing in the previous experiment. These pots also received 500 mL of dilute EM solution at fortnight intervals throughout the experimental period for mungbean (previous experiment) as well as for black grams (present experiment).
Treatments and experimental design
There were 6 treatments in each of the 4 soil amendment systems. These include:
i) Control (without any microbial inoculation).
ii) Effective Microorganisms (EM).
iii) B. japonicum st. MN-S. iv) B. japonicum st. MN-S + EM.
v) B. japonicum st. TAL- 102.
vi) B. japonicum st. TAL- 102 + EM.
Black gram seeds were surface sterilized with 1.0% sodium hypo- chlorite solution followed by several washings with sterilized water. Seeds were soaked in sterilized water for 2 h and left in covered petri plates over night to facilitate rapid and uniform germination. Seeds for respective B. japonicum treatments were pelted with peat based single strain inocula of B. japonicum st. TAL- 102 and B. japonicum st. MN-S with concentrated sugar solution as an ad- hesive. Initially 4 seeds were sown in each pot, which were thinned to 2 uniform seedlings on emergence. Each treatment was replica- ted 3 times. Pots were arranged in a completely randomized design on a bench in a wire netting house under natural conditions of light and temperature. Plants were irrigated with tap water of good quality whenever required.
Data collection and statistical analysis
Plants were harvested at flowering and maturity stages. The data regarding shoot length, root and shoot biomass were recorded at both the harvesting stages while that of number and biomass of nodules were recorded only at flowering stage. Data regarding various yield parameters; pod number, pod length, number of seeds per pod and grain yield were recorded at maturity. All the data were analyzed statistically by applying ANOVA followed by Duncan’s multiple range test (Steel and Torrie, 1980) to separate the treatment means.
Effect of microbial inoculation on shoot and root growth
Analysis of variance shows that effect of B. japonicum (B) inoculation was significant for shoot length (Table 1). At flowering stage, effect of either of the two B. japonicum strains was not much pronounced. However, at maturity stage, both the B. japonicum strains markedly enhanced shoot length in un-amended and farmyard manure (FYM) amended soils (Table 2). Both the B. japonicum strains enhanced shoot biomass in NPK amended soil. Effect of B. japonicum st. MN-S was more pronounced and signi- ficant both at flowering stage and maturity (Table 2). Effect of B. japonicum inoculation was also significant for root biomass (Table 1). In green manure (GM) amended soil, both the B. japonicum strains markedly suppressed root biomass at maturity stage. By contrast, in NPK amended soil a significant increase in root biomass was recorded by inoculation of either of the two B. japonicum strains. In un-amended as well as in FYM amended soil, neither of the two B. japonicum strains exhibited pro- nounced effect on root biomass (Table 2).
Analysis of variance reveals the significant effect of EM application on shoot length and biomass as well as on root biomass. The interactive effect of EM and soilamendment (A) was also significant for shoot length. Similarly, the effect of A × B × EM was significant for root biomass (Table 1). The most pronounced and significant effect of EM application was observed on shoot dry bio- mass in NPK amendment. Similar effect of EM appli- cation on root dry biomass was also recorded in NPK amendment at maturity stage. Neither of the two B. japonicum strains showed significant response to EM application with respect to root and shoot growth in any of the 4 soil amendment systems (Table 2).
Effect of microbial inoculation on nodulation
Effect of soil amendments was significant both for num- ber and fresh biomass of nodules (Table 3). The highest nodules number was recorded in un-amended soil followed by NPK amendment. Both the organic amend- ments resulted in a marked suppression in nodules number. Adverse effect was more pronounced due to GM than FYM amendment (Table 4).
B. japonicum inoculation showed a significant effect on nodulation. Nodules number was significantly enhanced by both the B. japonicum strains in un-amended as well as in NPK amended soil. B. japonicum st. MN-S was more effective in un-amended soil while st. TAL-102 was more effective in NPK amendment. Effect of inoculationon nodules biomass was also much pronounced in these two types of soils. Conversely, inoculation of either of the two B. japonicum strains failed to show significant effect on number and biomass of nodules in GM and FYM amended soils (Table 4).
Effect of EM application on nodulation was variable with respect to soil amendments. Analysis of variance shows that the interactive effect of EM and soil amend- ments was highly significant (P 0.01 and 0.001) both for nodule number and biomass (Table 3). In FYM amended soil, EM application markedly enhanced nodule number both in B. japonicum inoculated and un-inoculated treat- ments. In contrast to that, in NPK amendment, EM application suppressed number as well as biomass of nodules in B. japonicum inoculated plants. In other soil amendment systems, effect of EM application on nodulation was not much pronounced (Table 4).
Effect of microbial inoculation on yield
Analysis of variance shows that effect of soil amend- ments was significant for various yield parameters; num- ber of pods per plants, pod length, number of seeds per pod and grain yield (Table 3). Generally, values of these parameters were lower in GM amendment as compared to other soil amendment systems (Table 4). B. japonicum st. MN-S enhanced number of pods per plant by 26% in un-amended soil. By contrast, both the species reduced pod number by 40% in GM amendment. However, all the effects were insignificant statistically. Effect of B. japo- nicum inoculation on pod length and number of seeds per pod was also insignificant in all the 4 soil amendment systems. Grain yield was significantly enhanced by 46% due to each of the two B. japonicum strains in NPK amended soil. EM application resulted in significant increase of 48% in grain yield in NPK amended soil. In general, effect of EM application on various yield para- meters was insignificant (Tables 3 and 4).
In the present study, suitability of cross inoculation of two B. japonicum strains; TAL-102 (soybean isolate) and MN- S (mungbean isolate) inoculation to mashbean for bather growth, yield and nodulation characteristics was studied in different soil amendment systems. Both the strains proved suitable for mashbean. However, the affectivity of the two inoculated strains was associated with the type of soil amendment. Nodule number was significantly in- creased by inoculation of either of the two strains in NPK amendment. Nodule biomass was also markedly enhanced in inoculated treatments in this soil amendment system. As a consequence of improved nodulation, a similar significant improvement in grain yield was also evident due to inoculated B. japonicum strains. Earlier, Dubey (1998) obtained highest grain yield in soybean when host plant was inoculated with Bradyrhizobium in combination with NPK fertilizers. In the present study, response of nodulation to B. japonicum strains inocula- tion in un-amended soil was similar to that of response in NPK fertilizers amendments. However, unlike that of NPK fertilizer amendment, grain yield was not enhanced in un- amended soil in response to B. japonicum inoculation. Earlier, Mahmood and Athar (2008) reported that cross inoculation of mashbean with rhizobia isolated from Dalbergia sissoo, Leucaena leucocephala, Pithecel- lobium dulce, Prosopis cineraria, Prosopis glandulosa and Prosopis juliflora significantly enhanced dry weight and nitrogen contents of mashbean. In the present study, in both the organic matter amendments; farmyard and green manure amended soils, nodulation was very poor as compared to NPK amended and un-amended soils. Inoculation of both the B. japonicum strains failed to enhance nodulation in these two organic matters amended soils.
Effect of EM application on nodulation was variable in different soil amendment systems. EM application mar- kedly enhanced nodule number both in B. japonicum inoculated and un-inoculated treatments in FYM amen- ded soil. Conversely, in NPK amendment, EM application adversely affected the nodulation both in terms of number and biomass. In GM amendment as well as in un- amended soil, effect of EM application was not pronoun- ced. Earlier reports regarding the effect of EM application on nodulation are also contradictory. EM application caused a significant reduction in nodule number but increased the size and biomass of nodules in Trifolium alexand rinum (Bajwa et al., 1999b). By contrast, Javaid et al. (2000) noted a significant increase in nodulation in Vigna radiata due to EM application. Javaid et al. (2002) have reported similar effects of long-term EM application and organic manures on nodulation in Phaseolus vulgaris L. In seems probable that soil amendments as well as indigenous population of soil microorganisms determine the nodulation response of host plant to EM application.
Similar to that of nodulation, effect of EM application on plant growth and yield was also variable in different soil amendments. The most pronounced and significant effect of EM application on shoot and root dry biomass was observed in NPK amendment. Likewise, EM application resulted in 48% increase in grain yield in NPK amended soil without B. japonicum inoculation. In rest of the treat- ments, effect of EM application was not much pro- nounced. Earlier there are contradictory reports regarding the effect of EM application on crop growth and yield. Many workers have reported increase in crop growth and yield by the application of EM (Daly and Stewart, 1999; Yan and Xu, 2002; Javaid, 2006; Khaliq et al., 2006). However, the investigations of other workers have revealed that the effect of EM on crop growth and yield was usually not evident or even negative especially in the first test crop (Bajwa et al., 1995, 1999b; Diass et al., 2008; Javaid et al., 2008). Certain soil properties and the indigenous soil microbial populations are often con- straints to the establishment of EM cultures. Studies have shown that these constrains can be overcome through periodic repeated applications of EM at least during the first few years (Sangakkara et al., 1998; Javaid et al., 2000, 2002). According to Kinjo et al. (2000) the lack of consistency in results of the experiments regarding EM application may be due to variable cultural conditions employed in previous studies. Imai and Higa (1994) stated that the observed decline in crop yields could often be attributed to the fact that soils, where conventional farming is practiced, have become disease-inducing or putrefactive soils from long-term use of pesticides and chemical fertilizers. Consequently, it takes time to establish a disease-suppressive or zymogenic soil. Until this conversion process is completed, it is virtually impos- sible to exceed crop yields that were obtained with conventional farming methods. However, the present study reveals that the effect of EM application on crop growth and yield is associated with the type of soil amendment used. This study concludes that the benefits of B. japonicum strains TAL- 102 and MN-S and EM to black gram can be best exploited by applying these biofertilizers in NPK amended soils. However, further field trials are required before these findings are recommended to the farmers for field application of these biofertlizers.
Prof. Dr Tahir Hussain, Director Nature Farming Research Centre Faisalabad provided EM solution and Dr. Fouzia Yousaf Hafeez, NIBGE Pakistan provided B. japonicum cultures.
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Influence of "effective microorganisms" (EM) on vegetable production and carbon mineralization--a preliminary investigation
The influence of Effective Microorganisms (EM), a commercially available microbial inoculant containing yeasts, fungi, bacteria and actinomycetes, was evaluated in field trials of commercially produced, irrigated vegetable crops on "organic" farms in Canterbury, New Zealand during 1994-1995, and in a laboratory incubation. EM plus molasses were both applied, at 10 L ha-1 in 10,000 L ha-1 water, three times to the onions, twice to the peas and seven times to the sweetcorn. EM plus molasses increased the onion yield by 29% and the proportion of highest grade onions by 76%. EM plus molasses also increased pea yields by 31% and sweetcorn cob weights by 23%. A four week incubation at 30 degrees C of loamy sand and 1% w/w pasture litter had treatments including a control, glucose, and EM plus glucose, and captured respired carbon (C) using NaOH traps. By the end of the incubation the glucose treatment had respired 38% more C than the control. The EM treatment respired an additional 8% more C than the glucose treatment. Using EM stimulated C mineralization in the laboratory incubation, but a corresponding increase in mineralization of organic nitrogen, phosphorus and sulphur was not measured.
Journal Title: Journal of sustainable agriculture.
Journal Volume/Issue: 1999. v. 14 (2/3)
Main Author: Daly, M.J.
Other Authors: Stewart, D.P.C.
biological activity in soil
For more information, please visit the USDA National Agricultural Library.
Influence of Foliar Application by EM “Effective Microorganisms”, Amino Acids and Yeast on Growth, Yield and Quality of Two Cultivars of Onion Plants under Newly Reclaimed Soil
Z. F. Fawzy1, Abou El-magd M. M.1, Yunsheng Li2, Zhu Ouyang2 & A. M. Hoda1
1 Vegetable Research Dept., National Research Centre, Dokki, Cairo, Egypt
2 Yucheng Comprehensive Experimental Station, Key Laboratory of Ecosystem Network Observation and Modeling, Institute of Geographic Sciences and Natural Resources Research, Chinese Academy of Sciences, China
Correspondence: Yunsheng Li, Yucheng Comprehensive Experimental Station, Key Laboratory of Ecosystem Network Observation and Modeling, Institute of Geographic Sciences and Natural Resources Research, Chinese Academy of Sciences, China. E-mail: email@example.com
Received: June 21, 2012
This research was funded by the key project of IGSNRR, CAS, grant number: 2012SJ003
Two field experiments were carried out in two successive seasons of 2009/2010 and 2010/2011 in newly reclaimed soil at Wady Elmollak, Ismailia Governorate, Egypt to study response of two varieties of onion plant “Giza 20 and Super X” of foliar spraying of EM “Effective microorganisms”, amino acids and yeast on growth, and its quality as well as chemical composition. Results showed that Giza 20 cv gave the highest amount of vegetative growth “plant height and fresh weight of leaves” in the two seasons. Whilst, Super X cv. gave the highest amount of fresh weight of bulbs and whole plants. Moreover, using Super X cv gave the highest yield and quality on onion. Furthermore, Giza 20 cv. gave the highest amount of T.S.S, N, P and K% as well as some trace elements compared with Super X cv. With regard to foliar application treatments, the results indicated that, using EM, amino acids and yeast had positive promoting effects by providing supplemental doses of these components on growth, yield and its quality as well as all chemical composition compared with control plants. It may be concluded that using yeast at rates of 3 gm./L gives the highest growth parameters. However, using EM at rates of 3 cm/ L gives the highest yield and its quality of onion plants. Generally, it can be found that, using Super X cv. with foliar spraying of EM give the highest amount of growth, yield and quality of onion plants.
Keywords: onion, EM “effective microorganism”, amino acids, yeast, cultivars, growth, yield, chemical composition
Onion (Allium cepa L.) is an important bulb crop grown worldwide. It is an important crop in all condiment and used of flavouring the food, both at mature and immature bulb stages besides being used as salad and pickles. To lesser extent, it is used by processing industry for dehydration in the form of onion flakes and powder, which are in great demand in the world market. The great advances in techniques and methods of production of this crop in Egypt during the last decade perhaps open enormous pathways for exportation. Thus, the present work has involved studies on foliar application onion with EM (effective microorganisms) amino green and yeast because it is well known that this kind of treatments for obtaining clean and safe product.
Farmers have adopted the strategy of increasing crop yields by applying large amounts of chemical fertilizers and pesticides. At present, however, the negative effects of heavy applications of chemical inputs, in terms of production, environment, and quality deterioration are becoming apparent (Nishio, 1996). The ultimate goal of sustainable agriculture is to develop farming systems that are productive, profitable, energy-conserving, environmentally-sound, conserving of natural resources such as soil and water, and that ensure food safety and quality.
The cultivars genotype plays an important role of growth, yield and quality on onion plants (Mostafa, 1998; Daiz, 1994; Mohanty, 2001; Rahmanet al., 2002; Leilahet al., 2003; Gomaa, 2006; Haydar et al., 2007).
Microbial fertilizer is one way that organic farmers are able to increase yield and quality of crops without a large investment of money and labor (Pham, 2004). Moreover, microbial fertilizer can clean the environment and encourage the productive capacity of land by reducing the amount of chemical fertilizer consumption (Pham, 2004).
Various effective microorganisms and physiological active substances are contained in EM. Therefore, it must be effective to improve the quality and productivity of soil and to increase the vegetable production (Higa, 1991). In general, the vegetable crops require many nutrients for short period of cultivation. It seems that EM is a good supply source of nutrients in vegetable crops, because EM can make a favorable condition for the growth of crops, promoting the mobilization of non-soluble and activating the beneficial microorganisms in soil (Higa,. 1991, 2000; Hussain et al., 2002). Application of EM is known to enhance crop growth and yield in many vegetable crops (Kengo & Hui-lian, 2000; Sheng & Lian, 2002; Javaid, 2006, 2009; Khaliq et al., 2006; Daiss et al., 2008; Javaid & Nasir, 2010).
The requirement of amino acids in essential quantities is well known as a means to increase yield and overall quality of corps (Sanaa, et al., 2001; Slviero et al., 2001; Attoa et al., 2002; El- Shabasi et al., 2005; Awad et al., 2007; Fawzy, 2007; Al-Said & Kamal, 2008; Faten et al., 2010; Fawzy et al, 2010; Shaheen et al., 2010).
Dry yeast is a natural bio-substance suggested to be of useful stimulatory, nutritional and protective functions when it is applied on to vegetable plants during stressful condition due to its hormones, sugars, amino and nucleic acids, vitamins and minerals. The dry bread yeast (Saccharomyces cerevisiae) is a kind of the used biofertilizers in soil fertilization or in foliar application on the shoots of vegetable crops (El-Ghamriny et al., 1999). This is because it’s content of many nutrient elements and being productive compounds of semi growth regulator compounds like auxins, gibberellins and cytokinins (Glick, 1995). Glick recorded that the yeast was capable of increasing the simulative growth compounds like gibberellins, auxins and cytokinins that act in improving plant cell division and growth. Foliar application of yeast was found to increase growth, yield and quality of many vegetable crops (Abou El-Nasr et al., 2001; Gomaa et al., 2005, Mona et al., 2005; El-Tohamy & El-Greadly, 2007; El-Tohamy et al., 2008, Fawzy, 2007; Hussain & Khalaf, 2007; Fawzy et al., 2010; Ghoname et al., 2010).
The aim of present study was to evaluate the efficiency of foliar application of EM, active yeast extract and amino acids on improving onion growth, productivity and bulbs quality.
2. Material and Methods
Two successive field experiments were carried out on onion (Allium cepa, L.) in newly reclaimed soil at Wady Elmollak, Ismailia Governorate, Egypt during 2009/2010 and 2010/2011 seasons to study response of two varieties of onion plant growth, yield and quality to foliar nutrients. Seeds of onion cvs. Giza 20 and Super X were sown in the nursery on 15th and 7th of October in 2009 and 2010 seasons, respectively. All required agricultural managements for seedling production were carried out. Seedling was transplanted in the field after 45 days.
The experimental design was split plot with three replicates. Varieties of onion accepted the main plot and treatments of foliar nutrients were randomly allocated in the sub plots. Each experiment included twenty treatments representing the interaction between two varieties of onion i.e., Giza 20 and Super X with ten rates of foliar nutrients as follows:
* Control (Spray with tap water).
* Foliar spray of EM = Effective microorganism, at three rates (1 cm/L, 2 cm/L and 3 cm/L).
EM solution consists of useful micro-organisms and it safe for plants and soil. EM consists of photosynthesis bacteria, lactic acid bacteria and yeast. An EM effective microorganism is bio-fertilizer was got from Ministry of Egyptian Agriculture.
* Foliar spray of amino green II compound at three rates (1 cm/L, 2 cm/L and 3 cm/L.)
Amino green compound contains (w/v) total organic acids plus amino acids 15 %, iron (Fe), 2.9 %, Zinc (Zn) 1.4 % and Manganese (Mn) 0.7 %. Free amino acids, proline, Hydroxyl proline, Glysine, Alanine, Faline, Methionine, Escoliosin, Lyosine, Cyctin, Finel Alanin, Syrin, Glotomic) Arjenin, Hydroxy, Lysing and Hystiden. (Compound contain organic and amino acids and trace elements was got at from Dishner company for chemicals and trading, Giza- Egypt).
* Active dry yeast was used at three rates (1 gm. /L., 2 gm. /L., and 3 gm. /L)
The analysis of dry yeast was protein (47.2 %), Arginine (2.6 %), Glycin (2.6 %), Histidine (1.4 %), Esoloysine (2.9 %), Leucine (3.5 %), Lycine (3.8 %), Mehioninecystine (0.6 %), Phenyl-alanine (3 %), Tryptophan (0.5 %) and vitamin B (2.9 %) (N. R. P. 1977), Goyal and Khuller (1992), Yatskovskaya et al. (1992), Ahmed et al. (1997) and Khedr and Farid (2002) reported that yeast preparation contained carbohydrates, sugars, hormones, macro and micro elements in suitable balance.
Plants were sprayed with dry yeast, amino green compound and EM solution every week after month of transplanting for two month.
A. Vegetative growth: five plants from each plot were chosen randomly 120 days after transplanting for measuring the following characters:
1. Plant height (cm).
2. Leaves number/ plant.
3. Bulb diameter (cm).
4. Neck diameter (cm).
5. Bulbing ratio
6. Fresh weight of plant and its organs (leaves and bulbs).
B. Yield: total yield of onion was recorded per feddan.
C. Bulb quality: weight and diameter of bulbs at harvesting were recorded.
Total soluble solids (T.S.S) were assayed according to A.O.A.C (1990). Total nitrogen and phosphorus contents were determined using Kieldahl method and colorimetric method using spectrophotometer (SPECTRONIC 20D, Milton Roy Co. Ltd., USA), according to the procedure described by Cottenie (1980). Potassium content was measured using flame photometer method (JENWAY, PFP-7, ELE Instrument Co. Ltd., UK) as described by Chapman and Pratt (1982).
Copper, Zinc, Iron and Magnesium were determined using the Atomic Absorption Spectrophotometer (Philips) according the methods described by Chapman and Pratt (1961).
The obtained data were subjected to the analysis of variance procedure and mean were compared using the L.S.D. method at 5 % level of significance according to Gomez and Gomez (1984).
3. Results and Discussion
3.1 Vegetative Growth
3.1.1 Effect of Varieties
Vegetative characters of onion plant expressed as plant height, leaf numbers, fresh weight of leaves, bulbs and total plant were statistically affected by varieties of onion plants (Table 1). The differences resulting by the varieties reached the level of significant at all vegetative growth characters in the two seasons of study except for number of leaves in the second seasons. In addition, the highest values in vegetative growth were obtained by the Giza 20 cv. except for fresh weight of bulb. On the other hand, the lowest values of plant growth (plant height, leaf number and fresh weight of leaves) were recorded by Super X cv. The observed differences in vegetative growth of cultivars are mainly due to the genotype of each cultivar. The observed differences in vegetative growth of cultivars are mainly due to the genotype of each cultivar. This result was in harmony with previous findings (Daiz, 1994; Mohanty, 2001; Rahman et al., 2002; Haydar et al., 2007).
3.1.2 Effect of Foliar Application
Data in Table 2 shows clearly that foliar application of EM, amino green and yeast have a significant effect on the vegetative growth characterizes of onion plant in the two seasons of study. The increments were gradually and consistently with increasing the levels of yeast 1 gm. to 3 gm. /L. Generally, it could be concluded that, the highest values of plant height, number of leaves and fresh weight of onion plants were recorded by using yeast at rates of 3 gm. /L. compared with other treatments. On the contrary, the lowest values of vegetative growth of onion plants recorded by control plants (foliar spray with water). These findings were true in both seasons. These increments with using yeast as a foliar spray might be attributed to the effect of yeast extract in increasing levels of endogenous hormones in treated plants which could be interpreted to cell elongation and cell division (Khedr and Faried, 2002). Also, these results may be due to the physiological roles of vitamins and amino acids in the yeast extract which increased the metabolic processes rate and levels of indigenous hormones, i.e. LAA and GA3 (Chailakhyn, 1957; N.R.P., 1977).
3.1.3 Effect of the Interaction
The interaction effect within varieties of onion plants and foliar application treatments on vegetative characters of onion plant expressed as plant height leaves number, fresh weight of leaves, bulbs and total plant of onion plants shown in Table 3. There were significant differences in the all vegetative growth parameters in the first season except for number of leave parameter. Whilst, there were no significant effect in the all vegetative growth parameters in the second season except for total fresh weight of whole onion plant. Moreover, the highest plant height was recorded by Giza 20 cv. and foliar spray with yeast at a rate of 1 gm. /L in the first season and by Giza 20 cv. with foliar spray of yeast at rates of 3 gm. / L in the second one. On the contrary, the lowest value of plant height was found by Giza 20 cv. with foliar spray of water in the first season and by Super X cv. with control treatment (foliar spray with water) in the second season. With regard of number of leaves, data in Table 3 show that, the highest amount number of leaves was recorded by using Giza 20 cv with foliar spray of yeast at a rate of 3 gm. /L in the first season and by Super X cv and foliar spray of EM at rates of 3 cm/L in the second season. On the other hand, the lowest amount of number of leaves was found by Super X cv. and foliar spray of water in the first season and with Giza 20 cv. and foliar spray with water in the second season, respectively. However, the highest fresh weight of leaves was recorded by Giza 20 cv. with foliar spray of yeast at rates of 3gm. /L in the first season, whilst, by using Giza 20 cv with foliar spray of AG at rates of 3 cm/L in the second one. On the contrary, the lowest values were recorded by Super X cv. with foliar spray of water. These findings were true in both seasons. Moreover, data in Table 3 demonstrated that, the highest fresh weight of bulb and whole plants were recorded by using Super X cv. with foliar spray of EM at rates of 3cm/L. On the other hand, the lowest values were found by Giza 20 cv. with foliar spray with water. These findings were true in both growing seasons.
3.2 Yield and Its Quality
3.2.1 Effect of Varieties
Data in Table (4) illustrated that, there were significant differences in the bulb quality (bulb diameter, neck diameter, bulbing ratio and bulb weight) and total yield between the different varieties of onion plants. Moreover, the highest bulb diameter, bulb weight and total yield of onion were produced by Super X cv. On the contrary,
the lowest bulb diameter, bulb weight and yield of onion plants were recorded by using Giza 20 cv. These findings held good in both two experimental seasons. With regarding of neck diameter and bulbing ratio of onion. Results in Table (4) show that, the highest neck diameter and bulbing ratio of onion bulb were recorded by Giza
20 cv. and the lowest value of neck diameter and bulbing ratio found by super X cv. These results might be correlated with the gene action of the tested cultivars. These results are agreed with those obtained by Mostafa (1998), Mohanty (2001), Rahman et al. (2002), Leilah et al. (2003), Gomaa (2006), Haydar et al. (2007).
3.2.2 Effect of Foliar Application
Data presented in Table (5) show that, foliar application of EM, amino green, and yeast increased all bulb quality (bulb diameter, neck diameter, bulbing ratio and bulb weight) and total yield of onion compared with control. However, the highest bulb quality (bulb diameter, neck diameter, bulbing ratio and bulb weight) and total yield of onion plants was recorded by foliar application of EM at rates of 3 gm. /L. in the two seasons of study except for bulb diameter in the two seasons of study and bulb weight in the second season without any significant with foliar spray of EM at rates of 3 gm./L. On the contrary, the lowest bulb quality (bulb diameter, neck diameter,
bulbing ratio and bulb weight) and total yield of onion plants was recorded by foliar spray with water (control). The superiority of bulb quality and total yield of onion plants by using EM may be attributed to microorganisms improve crop growth and yield by increasing photosynthesis, producing bioactive substances such as hormones and enzymes, controlling soil diseases and accelerating decomposition of lignin materials in the soil (Higa, 2000; Hussain et al., 2002).
3.2.3 Effect of the Interaction
The interaction effect within varieties of onion plants and foliar application treatments on bulb quality (bulb diameter, neck diameter, bulbing ratio and bulb weight) and total yield are shown in Table (6). The obtained data reveals that, the interaction treatments significantly affected all bulb quality and total yield in the two seasons of study. These results held good in the two experimental seasons. Generally, it could be summarized that, the highest amount of bulb diameter was recorded by Super X cv. with foliar application of yeasts at a rate of 3gm/L. On the contrary, the lowest bulb diameter recorded by Giza 20 cv. with foliar spray with water only (control). The highest neck diameter of bulb recorded by Giza 20 cv. with foliar spray AG at rates of 3 cm/L. On the contrary, the lowest values of neck diameter of bulb recorded by Super X cv. with foliar spray of water only (control). With regard of bulbing ratio. Data in Table (6) show that the highest bulbing ratio recorded when using Giza 20 cv. receiving foliar spray with EM at rates of 2 cm/L. On the contrary, the lowest amount of bulbing ratio recorded by using Super X cv. receiving foliar spray of water only (control) in the first season and by Super X cv. receiving foliar spray of AG at rates of 1 or 2 cm/L in the second one. With regard of bulb weight, data in Table (6) show that the highest bulb weight and total yield of onion plants were recorded by Super X cv with foliar spray of EM at a rate of 3 cm/ L. Whilst, the lowest amount of bulb weight and total yield of onion plants were found by using Giza 20 cv and foliar spray with water. These results held good in the two seasons of study.
3.3.2 Effect of Foliar Application
Data in Table (7) show clearly that foliar application of EM, amino green and yeast had a significant effect on the chemical content characterizes of onion tissues in the two seasons of study. Generally, it could be concluded that, the highest values of T.S.S was recorded by using yeast at rates of 3 gm. /L. compared with other treatments. On the contrary, the lowest value of T.S.S was recorded by control plants (foliar spray with water). These findings were true in both seasons. With regards of N, P and K %, results in Table (7) demonstrated that, the highest values of N, P and K% were recorded by using foliar application of AG with different rates compared of other treatments. On the other hand, the lowest value of N, P and K % were recorded by control plants (foliar spray with water). These findings were true in both seasons of study.
This superiority might be due to that amino green compound contains many amino acids as well as some growth regulators and vitamins which stimulate and enhance the metabolism processes in plant tissues. Whereas, the previous studies have proved that, amino acids, can directly or indirectly influenc the physiological activities of the plants (Ei-Shabase et al., 2005; Awad et al., 2007; Al-Said & Kamal, 2008; Faten et al., 2010; Shaheen et al., 2010).
3.3.3 Effect of the Interaction
The interaction effect within varieties of onion plants and foliar application treatments on T.S.S, N, P and K % are shown in Table 8. The obtained data reveals that, the interaction treatments significantly affected T.S.S, P% in the two seasons of study and K% in the second season only. These results held good in the two experimental seasons. Generally, it could be summarized that, the highest amount of T.S.S, N, P and K % were recorded by using Giza 20 cv. with foliar application of EM and AG compared with foliar spray of yeast and control treatment (foliar spray of water). On the contrary, the lowest T.S.S, N, P and K % were recorded by Super X cv. with foliar spray with water only (control).
3.4 Trace Element Content
3.4.1 Effect of Varieties
Trace element content of onion tissues expressed as Fe, Cu, Zn and Mn were affected by varieties of onion plants (Figure 2). The differences resulting by the varieties reached the level of significant at Fe, Cu and Zn in the first season and Zn and Mn in the second one. Moreover, the highest values of all trace elements content were obtained by the Giza 20 cv. except for Mn in the first season and Fe in the second season, the highest amount was recorded by Super X cv. On the other hand, the lowest values were recorded by Super X cv. except for Mn and Fe ppm in the first and second seasons, respectively. The lowest values were found by Giza 20 cv. These results were true in the true seasons of study. These results were coincided with those reported by Gomaa (2006) and Haydar et al. (2007).
Figure 2. Effect of two varieties of onion on the Fe, Cu, Zn and Mn ppm in the 2009/2010 and 2010/2011 seasons
3.4.2 Effect of Foliar Application
Data in Table (9) show clearly that foliar application of EM, amino green and yeast had a significant effect on the trace elements content of onion tissues in the two seasons of study except for Mn ppm in the first season failed to significant effect. Generally, it could be concluded that, the highest values of trace elements content (Fe, Zn and Mn) were recorded by using foliar application of AG at rates of 3 cm/L. compared with other treatments. Whilst, the highest amount of Cu ppm was recorded by foliar spray of yeast at a rate of 3 bm./L. These results were true in the two seasons of study. On the contrary, the lowest value of all trace element measures were recorded by control plants (foliar spray with water). These findings were true in both seasons. These results may be due to the content of macro and micro elements of the dry yeast and amino green compound. El-Fouly 1983 reported that, foliar application of microelements is highly recommended under Egyptian conditions. In view of the fact the soil pH exceeds 7.5 and sometimes even 8.5 some areas show high CaCO3 contents which among other factors; make soil application of micronutrients more costly and unpractical. The trends of obtained results are in good accordance of the previous investigators such as (Ei-Shabase et al., 2005; Awad et al., 2007; Al-Said & Kamal, 2008; Faten et al., 2010).
3.4.3 Effect of the Interaction
The interaction effect within varieties of onion plants and foliar application treatments on trace element contents (Fe, Cu, Zn and Mn) are shown in Table 10. The obtained data reveals that, the interaction treatments had no significantly affected of all trace element content except for Fe in the first season. These results held good in the two experimental seasons. Generally, it could be summarized that, the highest amount of Fe ppm was recorded by using Giza 20 cv. with foliar application of AG at a rate of 3 cm/L in the first season and by Super X cv. with foliar spray of AG at a rate of 3cm/L in the second one. With regard of Cu ppm, the highest amount of Cu ppm was recorded by using Giza 20 cv. with foliar application of yeast at a rate of 3 gm. /L in the first season and by Super X cv with foliar spray of yeast at a rate of 3gm. /L in the second season. Meanwhile, the highest amount of Zn and Mn ppm were recorded by using Giza 20 cv. with foliar spray of AG at a rate of 3 cm/L. These findings were true in both seasons. On the contrary, the lowest amounts of all trace elements were recorded by Super X cv. with foliar spray with water only (control) except for Fe and Cu in the second season, the lowest amount was found by Giza 20 cv. with foliar spray with water only (control).
It can be recommended that all applied bio-stimulants have a positive and growth promoting effects on two cultivar of onion plants by providing supplemental doses of bio-stimulants “yeast, EM and amino acids”.
Generally, it can be concluded that, using Super X cv. with foliar spraying of EM gives highest amount of growth, yield and quality of onion plants.
The authors would express their thanks for all fruitful efforts presented and supported by Postdoctoral Program for African Researchers of China-Africa Science and Technology Partnership Program (CASTEP).
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Effective Microorganisms and Their Influence On Growth And Yield Of Pigweed (Amaranthus dubians)
Chrispaul Muthaura, David M. Musyimi, Joseph A. Ogur and Samuel V. Okello
Department of Botany and Horticulture, Faculty of Science, Maseno University, Maseno, Kenya
Pigweed (Amaranthus dubians) plants grow widely in many parts of the world. It is a very popular indigenous wild vegetable and is usually utilized by many communities as a nutritional additive. Pigweed is becoming very popular nowadays because of its high nutritive value and simple growth habits. However there is very little information about growth of this plant, using effective microorganisms, thus this study is designed to evaluate the effect of inoculation of effective microorganism on growth and yield of pigweed. The experiment was performed in five liter pots representing various conditions in the field. It comprised of four treatments, replicated, five times and arranged in a complete randomized design. One set of the treatments consisted of soil collected from the field, while the other treatments consisted of soil and organic manure prepared using effective microorganisms, sterilized soil treated with effective microorganisms, and sterilized soil plus organic manure without application of effective microorganisms respectively. Shoot height, stem diameter, leaf number per plant, leaf area, leaf fresh weight, leaf dry weight, root fresh weight, root dry weight and chlorophyll a and b contents were determined. Inoculated pigweeds with effective microorganisms recorded highest values in all the parameters measured except the root dry matter accumulation. There were significant differences (p≤0.05) in shoot height growth, stem diameter, leaf numbers per plant, leaf area, leaf fresh and dry weight and root fresh and dry weights among treatments. There were no significant differences in leaf chlorophyll content among the treatments even though chlorophyll a and b contents were slightly higher in plants inoculated with effective microorganisms. The three other treatments had significantly lower values of the parameters determined. The results from this study demonstrated that growth and yield of pigweeds may be improved by inoculating the plants with effective microorganisms, and as a result reduce the use of fertilizers in production of this vegetable hence promoting sustainable agriculture. More studies would be needed to determine the effects of effective microorganisms’ inoculation on other amaranthus species.
Keywords: pigweed, chlorophyll content, effective microorganisms, growth, yield, plant nutrition.
Environmental protection is getting more important for the agrarian, because of the purpose of sustainable agriculture. It is an important issue whether the replacement of organic fertilizers and chemical fertilizers with biofertilizers causes a beneficial increase in dry weight (Lévai et al., 2006). Nitrogen and phosphorus are essential nutrients for plant growth and development. Application of organic matter positively affects the growth and development of plant roots and shoots (Ghosh et al., 2004). There is evidence from the literature that application of biofertilisers can help to convert nutritionally important element from unavailable form to available form through biological processes. Organic fertilizers are good sources of nutrients for crop production and improving physical and chemical properties of soil. Microorganisms are important attributes in agriculture to promote the circulation of plant nutrients and reduce the need for chemical fertilizers. Biofertilizers are organic products containing living cells of different types of microorganisms that have emerged as important component of the integrated nutrient supply system and hold a great promise to improve crop yields through environmentally better nutrient supplies. These non-infecting plant growth promoting bacteria (PGPB) might affect mineral nutrition of plants through their influence on: i) growth, morphology and physiology of roots; ii) the physiology and development of plants; iii) the availability of nutrients; and nutrient uptake processes. A lot of microorganisms, for example species of Bacillus and Pseudomonas have a direct effect on the plant growth (Kloepper et al., 1986). The concept of effective microorganism, (EM) was developed by Professor Teruo Higa University of the Ryukyu, Okinawa, Japan (Higa, 1991). Effective microorganisms (EM) consist of mixed cultures of beneficial and naturally occurring microorganisms that can be applied as inoculants to increase the microbial diversity of soil and plant. Research has shown that the inoculation of EM cultures to the soil plant ecosystem can improve soil quality, soil health and the growth, yield and quality of the crops (Kengo and Huilian, 2000).
Decline in soil fertility, increased soil erosion and increasing shortage of food are major factors affecting human health in Africa. Fertilizers are costly and therefore out of reach of most resource poor farmers. There is also increasing evidence that synthetic agrochemicals and fertilizers have caused adverse effects on the environment leading to loss of biodiversity. This observation has promoted the need of introducing methods of farming aimed at reducing health risks including the use of effective microorganisms in organic farming. Inoculation of crops with effective microorganisms can improve vegetable crop yields and there by improve food security. Although pigweed is a very important indigenous vegetable in Kenya, there is hardly any information from the literature on the cultivation of pigweed using effective microorganisms.
Amaranth species are cultivated and consumed as leafy vegetables in many parts of the world (Schippers, 2000). Amaranth greens, are common leafy vegetables throughout the tropics and in many warm temperate
regions .The vegetables are very good sources of vitamins such as vitamin A, vitamin B6, vitamin C, riboflavin, and folate, and dietary minerals including calcium, iron, magnesium, phosphorus, potassium, zinc, copper, and manganese. Because of its valuable nutrition, some farmers grow amaranth today. Traditional green vegetables occupy an important role in household nutrition throughout the world particularly in rural areas.
Indigenous vegetables such as pigweed can contribute substantially to food security in the developing world and therefore need to be fully integrated in our farming systems. The vegetables are very rich in vitamins and proteins and, can help boost immunity amongst the HIV victims, improve the health of infants and breastfeeding mothers.
Effective microorganisms can contribute significantly to the production of indigenous vegetables such as pigweeds. Understanding and use of effective microorganisms in vegetable production in Kenya is limited (Muthaura, Personal communication). Data is lacking from the literature review to justify the use of effective microorganisms in cultivation and production of pigweeds in Kenya. The main objective of this study was
to investigate the influence of effective microorganisms on growth and yield of pigweed (Amaranthus dubians). It was hypothesized that organic manure prepared using effective microorganisms improves growth and yield of pigweed.
MATERIALS AND METHODS
Compost preparation using effective microorganisms A mixture of EM-1 (1 ml L-1), molasses and water was sprayed on rice straw and left to stand for about three weeks. The resultant manure was used as a media for pigweed propagation.
Experimental materials and growth conditions This experiment was conducted at Botanical Garden, Maseno University, Kenya. Seeds were acquired from the Botanical Garden of Maseno University. Ten viable seeds were sown in 5 liter plastic pots (20 cm diameter and 30 cm high), containing about 2.5 kg of sterilized soil (classified as kandiudalfic Eutrodox (Musyimi et al., 2007). The seedlings that emerged were thinned to two plants per pot, ten days after germination. The pots were watered on daily basis to maintain the soil moisture content at approximately 60% water holding capacity.
Experimental designs and treatments The experimental design was a completely randomized design with four treatments: Soil plus manure containing effective microorganisms (A), soil only (B), Soil plus manure only (C), and soil plus effective microorganisms without manure (D). The treatments were replicated five times and the pots containing the seeds were arranged inside a green house whose conditions were:
Temperature: min/max 20/410C and relative humidity: min/max 50/95% (Plate-1). Weeds were controlled by hand pulling. Watering was carried out on daily basis. After germination the seedlings were thinned out leaving only two plants per pot. Data collection commenced fourteen days after seed germination and seedling establishment.
Plate-1. Experimental layout in the greenhouse.
Measurement of growth parameters
Growth measurements commenced fourteen days after seed emergence and seedling establishment. Data was collected at an interval of four days up to the end of the experiment. Shoot height was measured using a meter rule. The number of fully expanded mature leaves was established by counting. Leaf area expansion was determined at the end of the experiment according to Jose et al. (2000). AL = 0.73 (LLX WL), where LL is the leaf length and WL is the maximum width measured for all leaves on each plant. The stem diameter was determined at the end of the study by use of a vernier caliper. At maturity, the plants were harvested and their roots, shoots and leaves separated. Roots were washed in tap water after soaking, blotted dry on paper towels and weighed using an electronic balance. Fresh weight reading for the roots shoots and stems were taken immediately after harvesting. All the plant samples were oven dried at 60 0C to constant dry weight for at least 48 hours after which their dry weight was taken using a weighing balance.
Determination of chlorophyll content
The fifth leaf from the shoot apex from each plant in each treatment was collected for chlorophyll extraction and concentration determination.
Chlorophyll concentration was determined in 80% acetone extract by use of a spectrophotometer. Absorbency was measured against an 80% acetone blank at 647nm and 664nm. The values of chlorophyll a, chlorophyll b was determined according to Yadegari et al. (2007) using the following formula:
Chlorophyll a = 13.19A664-2.57A647 (mg/g fresh weight)
Chlorophyll b = 22.1 A647-5.26 A664 (mg/g fresh weight)
Where, A664 was absorbance at 664nm and A647 was absorbance at 647nm, respectively.
Data collected was analyzed using SAS statistical computer package. Analysis of variance (ANOVA) was carried out to determine whether there were any significant differences among treatments on parameters
Shoot growth occurred in all the treatments over the experimental period. There were significant differences in shoot growth among the treatments (P≤0.05).
The treatment containing Effective microorganisms plus manure had the highest shoot growth compared to all other treatments, about 46% growth increase compared to the control treatment (soil only)
Effective microorganisms’ inoculation improved leaf production over the experimental period (Table-1). There were significant differences (p≤0.05), in leaf formation among treatments throughout the study period.
Plants inoculated with EM and organic manure recorded the highest number of leaves compared to all other treatments, this was about 26% of control plants.
Leaf expansion occurred in all the treatments. There were significant differences (p≤0.05), in leaf area growth at the end of the experiment (Table-2). Plants inoculated with EM manure recorded the highest leaf area
growth, followed by the treatments containing soil + manure about 35% of the control treatment (soil only).
Stem diameter growth increased among all the treatments. There were significant differences in stem diameter growth (p≤0.05); (Table-2). Plants inoculated with EM had the highest stem diameter growth, about
45.4% of the control plants (soil only). Leaf fresh weight: Leaf fresh weight increased in all the treatments over the experimental period, however there were significant differences (p≤0.05), among treatments. Plants inoculated with EM manure recorded the highest fresh weight, which was about 76% of the control plants (soil only).
Root fresh weight
Maximum root fresh weight was recorded in media having manure treated with EM. There were significant differences among the treatments for this parameter. Fresh weight was highest in soil inoculated with effective microorganisms.
Leaf dry weight
Plants inoculated with EM with soil and manure recorded the highest dry weight (Table-2), about 180% of the control plants (soil only). There were significant differences (p≤ 0.05) among all the treatments.
Root dry weight
Root dry mass was highest in soil without organic manure inoculated with effective microorganisms (Table- 2). There were significant differences (p≤0.05), among the treatments.
There was no significant change in the chlorophyll a and chlorophyll b synthesis among the treatments. However pigweeds inoculated with EM manure had relatively higher chlorophyll a and chlorophyll b content compared to all other treatments (Table-3). Chlorophyll a was also higher than chlorophyll b in all the treatments.
The results from the study indicate that inoculation of pigweeds with effective microorganisms increased the growth of shoot height, stem diameter, leaf number, leaf area, leaf fresh and dry weights, and root fresh and dry weights. Increased shoot height stem diameter growth probably reflects allocation of resources into shoots rather than roots (Tables 1 and 2). Increase in the number of leaves and leaf area are common
occurrences in plants that are provided with proper nutrition and this can increase the photosynthetic activity of the plants.
Increase in leaf area and number of leaves should result to higher rates of photosynthesis hence increased plant growth. For plants, a high rate of net carbon assimilation can result in higher biomass accumulation,
favouring future growth and reproduction. The position and distribution of leaves along the shoot influences the sink strength of the plants. During early stages of leaf growth, synthesis of chlorophyll, proteins and structural compounds is high resulting in high catabolic rates to support energy needs by the plants. Inoculation of effective microorganism can increase the available nutrition for plant roots and improve photosynthesis.
Singh et al. (2003) reported that biological seed and mucilage yield of Isabgol could be increased with application of animal manure and integrated systems due to improved soil physical and chemical properties.
Accumulation of dry matter and its distribution into different plant components is an important consideration in achieving desirable economic yield from crop plants (Singh and Yadav, 1989). Chlorophyll a and b content increased in all the treatments, even though the plants inoculated with effective microorganisms had relatively higher chlorophyll contents (Table-3). Increase in chlorophyll a and b contents of the pigweed may contribute to increased photosynthetic activity. The synthesis and degradation of the photosynthetic pigments are normally associated with the photosynthetic efficiency of the plants and their growth adaptability to different environments (Beadle, 1993). Increase in leaf chlorophyll content could in turn lead to increased protein synthesis of the plants and this could have a direct consequence on the plant growth and photosynthesis (Hendry et al., 1987). Nitrogen is one of the essential nutrients involved as a constituent of biomolecules such as nucleic acids, coenzymes and proteins (Sharma et al., 1995), any deviation in these
constituents would inhibit the growth and yield of plants. Protein concentrations in plants tend to increase with fertility level of the growth medium (Grant and Bailey,
In general effective microorganisms seem to have direct impact on growth and yield of pigweeds. Previous studies have demonstrated a consistent positive response with the use of effective microorganisms in crop
production and indicate the potential of this technology to reduce fertilizer use and increase the yield and quality of crops (Higa, 1991).
The results show that inoculation of pigweeds (Amaranthus dubians) with effective microorganisms can improve their growth and yields. To prevent the environmental pollution from extensive application of
fertilizers, the effective microorganisms could be recommended to farmers to insure the public health and a sustainable agriculture.The data collected proves that the use of effective microrganisms can lead to higher amaranthus yield (Amaranthus dubians). Further research should be done to quantify the numerous effects of EM on growth and yield of other amaranthus species. The local community should be sensitized on the use of EM to improve farming and thus help alleviate poverty; this should be done through workshops and seminars.
Mr. Peter Olewe is appreciated for his technical assistance during the laying out of the experiments in the greenhouse. We are grateful to the Department of Botany and Horticulture for financial support to carry out this study.
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