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NAF International Working Paper Series

Year 2013 paper n. 13/02

Short-term effects of seed-dressing with associative-nitrogen-fixing bacteria on the establishment and early growth of maize in Zimbabwe
G. V. Nkomo
Department of Crop and Soil Sciences, Faculty of Agriculture and Natural Resources, Africa University. P.O Box 1320 Mutare, Zimbabwe Corresponding author Email: gvnkomo@gmail.com

F. Mapanda
Department of Soil Science and Agricultural Engineering, Faculty of Agriculture, University of Zimbabwe, P.O Box MP167, Mt. Pleasant, Harare, Zimbabwe

A.Z. Chiteka
Department of Crop and Soil Sciences, Faculty of Agriculture and Natural Resources, Africa University. P.O Box 1320 Mutare, Zimbabwe

W.T. Mutezo
Department of Crop and Soil Sciences, Faculty of Agriculture and Natural Resources, Africa University. P.O Box 1320 Mutare, Zimbabwe

The online version of this article can be found at: http://economia.unipv.it/naf/

Scientific Board

Maria Sassi (Editor) - University of Pavia Johann Kirsten (Co-editor)- University of Pretoria Gero Carletto - The World Bank Piero Conforti - Food and Agriculture Organization of the United Nations Marco Cavalcante - United Nations World Food Programme Luc de Haese - Gent University Stefano Farolfi - Cirad - Joint Research Unit G-Eau University of Pretoria Ilaria Firmian -IFAD Mohamed Babekir Elgali University of Gezira Firmino G. Mucavele - Universidade Eduardo Mondlane Michele Nardella - International Cocoa Organization Nick Vink - University of Stellenbosch Alessandro Zanotta - Delegation of the European Commission to Zambia

Copyright @ Sassi Maria ed. Pavia -IT naf@eco.unipv.it ISBN 978-88-96189-12-2


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Short-term effects of seed-dressing with associative-nitrogen-fixing bacteria on the establishment and early growth of maize in Zimbabwe G. V. Nkomo*1, F. Mapanda2, A.Z. Chiteka1 and W.T. Mutezo1 1. Department of Crop and Soil Sciences, Faculty of Agriculture and Natural Resources, Africa University. P.O Box 1320 Mutare, Zimbabwe 2.Department of Soil Science and Agricultural Engineering, Faculty of Agriculture, University of Zimbabwe, P.O Box MP167, Mt. Pleasant, Harare, Zimbabwe *Corresponding author Email: gvnkomo@gmail.com Abstract Productive agriculture needs a large amount of expensive nitrogenous fertilizers. Improving nitrogen use efficiency of crop plants is thus of key importance. Despite the various interventions such as use of cattle, wood, leaves and in some instance bird droppings as fertiliser in maize production farmers still face low yields. This is due to the poor status of the manure and also the low volumes applied to correct the inherent low levels of nitrogen in the soil. The rationale of the experiment was to assess effect of nitrokara biofertiliser on maize crop emergence and early growth response to inoculation with nitrokara and a compound fertiliser application at planting. The experiment was conducted in plots on a sandy clay loam soil, involving six treatments (nitrokara +300kg Compound D, nitrokara + 300kg Compound D + 75kg AN, nitrokara +300kg Compound D +150kg AN, nitrokara +300kg Compound D +225 AN, nitrokara +300kg Compound D + 300 kg AN and 0 nitrokara+300 Compound D +0 AN). Early maturing SC 403 maize (Zea mays) was inoculated with nitrokara and a compound mineral fertilizer at 300 kg/ha at planting while AN was applied at 45 DAP. Ten plants from each net plot were randomly sampled at 5, 7, 9 and 10 days after planting. At 6 and 8 weeks actively growing 6 plants were chosen at random from the net plots were harvested for biomass determination. There were no significant differences (P >0.05) in crop emergence between inoculated and non-inoculated treatments at 5, 7, 9 and 10 DAP. Crop emergence increased from a grand mean of 62.5% at 5 DAP to 97.5% at 10 DAP. There were no significant differences (P>0.05) on plant biomass on treatments 1 to 6 at 4 weeks after planting (P<0.05) as well as at 8weeks after planting (P=0.27).However there were differences on the grand mean of plant biomass at 6 weeks which was 0.226 while at 6 weeks it was 3.49. The increase in plant biomasses indicate that nitrokara has the ability to promote the growth of early maturing maize plant. However the ability of performance depends on the proper association between A caulinodans in nitrokara and maize crop. Key Words: biofertiliser, symbiotic, plant biomass, inoculated

1. Introduction Several authors have indicated that inoculation with plant growth promoting rhizobacteria (PGPR) results in increased germination and seedling emergence and modify growth and yield of various cereal and non-cereal crops (Freitas and Germida, 1992; Chen et al., 1994; Javed and Arshad, 1997; Biswas et al., 2000; Matiru and Dakora, 2004; Wang et al., 2007). In recent years, use of microbial inoculants to enrich N and other nutrients in soil has become a practice in most of the countries, as far as economical and environmental view points are concerned (Amutha and Kannaiyan, 2011). Seed treatment with bioinoculants increases the plant growth and also increases the colonization of beneficial microorganisms in root region of the plants (Chen et al., 1994). Plant growth promoting rhizobacteria (PGPR) is a group of free living soil bacteria, which have ability to promote growth and yield of crop plant by direct and indirect mechanism. There are generally two types of PGPR, one that colonises inside plant cells called intracellular PGPR (iPGPR) and the other that colonises outside plant in rhizosphere called extracellular PGPR (ePGPR)( Verma et al., 2010). The iPGPR and arbuscular mycorrhizal fungi (AM fungi) have three major components: the root itself which provides carbon in the form of sugars to the fungus, fungal structures within cortical cells of plant root that provide contact between fungus and the plant cytoplasm and the extra radical hyphae that aid uptake of nutrients and water (Smith and Read, 1997). Global concern for sustainable agriculture and minimization on the use of chemical fertilisers has led to the concept of integrated plant nutrient management which implies use of rhizospheric microorganisms along with judicious use of chemical fertilisers (Khokhar et al., 2002). Shevananda (2008) states that application of biofertilizers has become a great necessity to get yields of high quality and to avoid environmental pollution in Zimbabwe. One of most important factors that impact on the physiology of plants growth and development is the availability of nutrients which can be taken up by plants from soil. Nitrogen and phosphorous are important for plant growth, however plants have a limited ability to phosphorous from the environment, and thus need microbes involved in nutrient recycling (Mohammed, 2012).This is because phosphorous is highly immobile in soils. In a way, microorganisms serve as biofertilizers (El-kholy et al., 2005). Some authors have reported increased root, shoot weight with dual inoculation in maize while grain yields of the different maize genotypes treated with for example Azospirillum spp. varied between 1700 and 7300 kg/ha (Salmone and Dobereiner, 2004). Increasing yield was attributed to the plant growth promoting substances by root colonizing bacteria more than the biological nitrogen fixation

(Mohammed, 2012; Lin et al.,1983) stated that yield increased due to promoting root growth which in turn enhancing nutrients and water uptake from the soil. Maize seeds inoculated with rhizobacteria, Pseudomonas cepacia, P. fluorescens and Streptomyces aurantiacus in combination with 120 kg/ha of nitrogen increased crop yield up to 25% more than the non rhizobacterium colonized maize control (Adjanohoun et al., 2011).The effect of rhizobacterium-nitrogen combination was found to be 60% better than that of rhizobacterium alone (Shaharoona et al., 2006). Plant growth promoting rhizobacteria (PGPR) are believed to increase the Supply and or availability of primary nutrients to the host plant (Wu et al., 2005), promoting the synthesis of antibiotics, enzymes and fungicidal compounds (Bharathi et al., 2004; Jeun et al.,2004; Ahmad et al., 2006).

2. Materials and Methods 2.1 Study Site The experiment was conducted on the field at Africa University Farm (AU Farm). The soils at AU Farm are red Fersiallitic 5E soil under Zimbabwe soil classification system. The soils are sandy clay loam soils. Average annual rainfall ranges from 750-1200 mm while the altitude is at 1,131m above sea level. 2.2 Test Crop A very early maturity white maize variety (SC403, 61 days to silking and 113 days to physiological maturity) was used as the test crop. This hybrid variety is maize streak and mottle viruses tolerant, with a relatively short, flinty ear and excellent yield stability over a range of environments with relatively slow dry down rate (Seed-Co, 2012). In addition, it has outstanding drought tolerance with very good synchronization of silks with pollen. On average, SC 403 has a maximum yield potential of 5 t/ha and has outstanding tolerance to both Diplodia and Furasium cob rots (Seedco, 2012). SC 403 is widely recommended for harsh conditions where yields of less than 6t/ha are expected. 2.3 Treatments and Agronomic Management The experimental site was deep ploughed to about 25 cm depth, disked and harrowed to ensure attainment of a fine tilth. Experimental plots (4 m x 4 m) were laid out and arranged in a randomized complete block design with 4 blocks. The six treatments that were replicated twice in each block were: 1. Nitrokara and 300kg Compound D 2. Nitrokara + 300kg Compound D + 75kg AN 3. Nitrokara +300kg Compound D +150kg AN 4

4. Nitrokara +300kg Compound D +225 AN 5. Nitrokara +300kg Compound D + 300 kg AN 6. 0 Nitrokara+300 Compound D +0 AN Maize was sown at the spacing of maize used was 90 cm within rows x 50 cm between rows, with 2 plants per station, to make a population of 22 222plants/ha. Before planting maize seeds were dipped in the 5% sugar solution (3 spoonfuls in 1litre of water) first for 15minutes.The mixing ratio of Nitrokara was 100 g per 50kg of maize seed. 4 kg maize seed and 8g of Nitrokara were mixed with 1kg of soil and then mixed thoroughly with maize seed prior to planting. The trial area was kept pest and weed free always through hand and hoe weeding. Compound D was applied as basal fertilizer at a rate of 300 kg per hectare in all treatments. AN was applied at 45 days after planting (DAP).The plot size was 4 m x 4 m.

2.4 Sampling and Measurements Plant count was done on each net plot at 5, 7, 9 and 10 days after planting. Ten plants from each net plot were randomly sampled. At 6 and 8 weeks actively growing 6 plants were chosen at random from the net plots were harvested for biomass determination. Plants from the net plot were partitioned into leaves and stems. Dry mass was determined by oven drying the components at 60 0 C for 72 hours to constant weight before weighing.

2.5 Data Analysis Genstat Release 7.2 software was used to analyse the results. The least significant difference (P<0.005) method was used to separate the means.

3. Results There were no significant differences (P>0.005) on emergence % from 5days after planting, 7 days after planting, 9 days after planting as well 10 days after planting using maize seed inoculated with nitrokara. Emergence percentage varied with the number of days; at 5 days emergence % grand mean was 62.5%, at 7 DAP was 92.04 %, 9 DAP was 96.17% and 10 DAP was 97.54%. This is presented in the following graph 3.1.

Graph 3.1 showing different emergence % grand means from day 5 to day 10

Emergence percentage varied with the number of days and treatments; at 5days emergence % per treatment ranged from 61.88% to 62.88%, at 7 DAP was 91 % to 92.50%, 9 DAP was 95.75% to 96.62% and 10 DAP was 96.38% to 98.25%.There was a significant great increase from day 5 in emergence % up to day 7 and emergence percentage then becomes consistent up to day 10.This is presented in the following graph 3.2.

Graph 3.2 showing different emergence % means from treatment 1 to 6 from day 5 to day 10 6

There were no significant differences (P>0.005) on plant biomass on treatments 1 to 6 at 4 weeks after planting (P=4.07) as well as at 8weeks after planting (P=0.27).However there were differences on the grand mean of plant biomass at 6 weeks which was 0.226 while at 6 weeks it was 3.49.This is shown below in the tables.

Table 3.1 Biomass 1 and treatment means Biomass 1 Treatment 1 Treatment 2 Treatment 3 Treatment 4 Treatment 5 Treatment 6 Grand Mean Lsd Treatment Mean 0.153a 0.205a 0.229a 0.267ab 0.308bc 0.192ab 0.226 0.0786

Table 3.2 Biomass 1 and treatment means Biomass 2 Treatment 1 Treatment 2 Treatment 3 Treatment 4 Treatment 5 Treatment 6 Grand Mean Lsd Treatment Mean 3.32a 3.44a 3.45a 3.74a 3.80a 3.19a 3.49 1.307

Discussion Most of the maize crop had emerged after 7 DAP (92%).At 9 DAP most of the crop had emerged with an average emergence percentage of 97%. Treatments 4, 5 and 6 showed variations in biomass response to nitrokara. It is evident that plant growth promoting rhizobia (PGPRs) induced improved seed germination and plant development (Ahmad et al., 2006). The PGPR effect is generally beneficial to a wide range of crop production including maize (Esters et al., 2004). This positive effect of PGPRs could reasonably be attributed to induction of different vegetative hormones 7

displaying a well programmed pleitropic effects (Contesto et al., 2008). Thus specific PGPRs can be considered in maize production to improve field maize crop productivity. Similar results were reported on maize field trial with Azospirillium sp. treatment by Bashan and Holguin, 2004) and Cakmaki et al. (2006). Plant inoculation with Azospirillum sp. was found to induce significant changes of growth parameters such as plant biomass (Bashan and Holguin, 2004).

Conclusion The increase in plant biomasses indicate that nitrokara has the ability to promote the growth of early maturing maize plant. However the ability of performance depends on the proper association between A caulinodans in nitrokara and maize crop. Proper rhizobial inoculation is another important factor for seed germination, emergence and growth. This indicates that certain strains of rhizobia can promote growth of non-nitrogen fixing such as maize, possibly through mechanisms that involve changes in growth physiology and root morphology.

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