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Vol. 11(3), pp. 195-203, 2021
Copyright ©2021, the copyright of this article is retained by the author(s)
Influence of Arbuscular Mycorrhizal Fungi on (Zea mays) Cultivated on Sewage Contaminated Soil
Anozie, H.I; Wokocha, C.C; Madu, O.V
Department of Crop and Soil Science, Faculty of Agriculture
University of Port Harcourt, PMB 5323, Port Harcourt, Nigeria.
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Anozie, H. I
E-mail: henry.anozie@ uniport.edu.ng
Maize (Zea mays L.) is the world’s highest supplier of calorie with caloric supply of about 19.5%. Maize is the most important staple food in Nigeria and it has grown to be local ‘cash crop’ where at least 30% of the crop land has been devoted to small-scale maize production under various cropping systems (Ayeni, 1991). Introduced in Nigeria in the 16th century, maize is the fourth most consumed cereal ranked below sorghum, millet and rice (FAOSTAT, 2012). It has been recognized to be one of the longest ever cultivated food crops. Maize is also grown in several regions of the world and is referred to as the world best adapted crop (IITA, 2008). In Nigeria, the demand for maize is increasing at a faster rate daily (Sadiq et al., 2013).This may be due to the fact that the grain is being used for feeding poultry and also serve as the main food for many households (Ogunniyi, 2011). This has however, necessitated the need for research and more extension services to the rural famers not just to improve farm yield but to as well provide them with alternatives that will help them to continue production especially those living in areas where the land has been frequently polluted with sewage, hence the study.
Soil faces serious environmental and health challenge as a result of Heavy Metal (HM) contamination by Sewage pollution. AMF are essential for soil amelioration as they reduce the availability of HMs in the soil as a result of its inoculation due to significant increase in soil pH (Asrar and Elhindi, 2011).
Arbuscular Mycorrhizal Fungi (AMF) are widespread obligate symbiotic forming associations with 85% of the vascular plant species (Brundrett, 2018), dominating most of the tropical forest and temperate grassland ecosystems (Soudzilovskaia, 2018). Besides the fundamental role of AMF in plant nutrition and fitness, it is widely recognized that AMF have a substantial impact on ecosystem functioning. The AMF extra-radical mycelium also acts as an active distributor of carbon (C) in the soil, feeding soil heterotrophs (Pollierer, 2007) and stabilizing carbon in recalcitrant organic compounds (Sousa, 2012).
Soil contamination is one of the major environmental challenges affecting farmers and their crops. This however, arises as a result of improper sewage management, industrial waste mismanagement, and such like. Sewage contains some heavy metals that affect crop productivity (Sadiqet al., 2013). Once heavy metals have entered the soil, they have long-term effects on plants. Arbuscular Mycorrhiza are essential bio-agent, as they can significantly enhance the efficiency of the ecosystem by producing fungal structures like arbuscules and aid in the exchange of inorganic compounds and minerals required for the growth of plants, thereby providing considerable strength to plants (Babatunde et al., 2007). There is a symbiotic relationship between AMF and the plant roots which help to establish tolerance to HMs (Chandrasekaran et al., 2019). AMF has the ability to enhance the plant ability to uptake essential nutrients and partially exclude non-essential ones (Hashem et al., 2019). Several methods exist to clean up the environment from contaminants; however, many of them are costly or difficult to implement. Therefore, a cheaper and eco-friendly method is indispensable for amelioration of deleterious effects of sewage on soil. Regrettably, there is insufficient information on the influence of AMF on maize cultivated on sewage contaminated soil. Regrettably, there is insufficient information on the influence of AMF on maize cultivated on sewage contaminated soil. Therefore, this study was set up with the following objectives: to assess the influence of AMF on performance of maize cultivated on sewage contaminated soil and identify the best host among the three species of AMF used.
MATERIALS AND METHODS
The experiment was conducted at the Screen House, Department of Crop and Soil Science, University of Port Harcourt at Latitude 4054N and longitude 06055E with an average temperature of 27 0C, relative humidity of 78 % but decreases slightly in dry season and an average rainfall ranging from 2500 mm – 4000 mm per annum (Atijegbe et al., 2013).
The area has a biomass rainfall pattern with a long rainfall season between March and July and a short rainy season from September to early July after a short spell in August and a longer period from December to February (Akande et al., 2010)
Source of Seed Variety
The Maize variety used for this study was OBA SUPER 98, sourced from Agro-tropical Institute, Port Harcourt, Rivers State, Nigeria.
Source of AMF inocula
Three pure strains of Arbuscular Mycorrhiza Fungi namely Gigaspora specie, Glomus clarium and Glomus mossea gotten from the Department of Microbiology, University of Ibadan were used for this experiment.
Soil sampling and Sterilization
Samples of soil (0 – 30 cm depth) were randomly taken from the experimental farm for sterilization. Sterilization was done at a controlled temperature of 1210C for 6 hours and later allowed to cool for 72 hours. The sterilized samples were measured using a weigh balance and 10 kg was poured into each experimental bucket.
Sewage and AMF Application
Sewage measuring 750 ml was introduced to soil sample one week before planting and six weeks after planting respectively. After first sewage application, 50 g of Mycorrhiza inoculum was added to soil samples three days before planting.
Experimental Design and Planting
This experiment comprised of 2 factors (Mycorrhiza and Sewage) laid in completely randomized design (CRD) with 8 replications. Three seeds of maize were planted at 2.5 cm depth above soil surface in each bucket. Watering was done twice daily (morning and night) while weeding was done by handpicking at interval.
Data collection and analysis
Growth parameters such as Plant Height, Leaf Area, Stem Girth and Number of Leaves were collected biweekly. Data collected were analysed with the use of Gen Stat Software (GEN, 2012) and means were separated using Least Significant Difference (LSD) at 5 % significance level.
Soil Laboratory Analysis
Soil samples were analysed before planting and after planting. Soil particles size analysis was done using hydrogen method (Bouyoucos, 1962); Available P by Bray 1 method (Bray and Kurtz, 1945); Soil pH was determined in 1:1 (soil: water) ratio using a glass electrode pH meter. Macro kjedahl digestion distillation method was used in measuring Total Nitrogen. Total organic carbon was determined by the wet combustion method of Walkey and Black (1934) as modified by Juo (1979) in selected methods of soil analysis. Organic carbon was oxidized by potassium dichromate in the presence of concentrated sulphuric acid. Ferrous ammonium sulphate was then added and the excess black titrated with standard potassium permanganate. 1.0 g of representative soil sample was shaken in a conical flask with 50 ml of 1N- ammonium acetate for about 2 hours. The mixture was left over night and then filtered into plastic cups. The filtrate was used for determination of sodium, potassium, calcium and magnesium using Atomic Absorption Spectrophotometer (AAS). A 30 g of soil sample was digested in a mixture of concentrated nitric acid (HNO3), concentrated hydrochloric acid (HCl) and 27.5 % hydrogen peroxide (H2O2) according to the USEPA method 3050B for the analysis of heavy metals (USEPA, 1996). The extracts were analysed by atomic absorption spectrophotometer (Perin Elmer, Model No 2380)
Effect of various Species of AMF on selected Growth Parameters of Maize.
Table 1 show the effects of AMF on growth parameters of maize as follows:
Generally, the height value for the maize plant inoculated with my various species of Mycorrhiza was significantly higher than that of the Control in all weeks except in 10 WAP as shown in Table 1. In 4WAP, G.clarium recorded the highest plant height value of 34.2 cm among all the treatments while Gigaspora specie had the least 26.7 cm. However, G.clarium is statistically higher than Control (27.9 cm) and Gigaspora specie (26.7 cm) but the same with G.mossea (32.0 cm). In 8WAP, G.mossea recorded the highest plant height value of 92.5 cm while the Control had the least value 81.5 cm. Although G.mossea is statistically higher than the Control and Plants inoculated with other AMF specie, it is the same with Gigaspora specie (88.5 cm) and G.clarium (89.5 cm) but different from Control (81.5 cm). In 10WAP, Control recorded the highest plant height value, 90.1 cm while G.clarium recorded the least value, 85.8 cm. However, the Control is statistically the same as others.
At 4WAP, G.clarium recorded the highest area value (49.8 cm2) while G.mossea recorded the least value 45.0 cm2. However, there is no statistical difference among treatments. At 8WAP G.mossea recorded the highest value (320 cm2) while Control recorded the least value (187 cm2). However, G.mossea (320 cm2) is only significantly higher than Control (187 cm2) but the same with G.clarium (258 cm2) and Gigaspora specie (261 cm2). Also, the Control (187 cm2), G.clarium (258 cm2) and Gigaspora specie (261 cm2) are statistically the same. At 10WAP, G.clarium recorded the highest value (316 cm2) while Gigaspora specie (227 cm2) recorded the least. However, G.clarium (316 cm2) is statistically higher than Gigaspora specie (227 cm2) but shows no statistical difference with the Control (291 cm2) and G.mossea (312 cm2).
Number of Leaves
Generally, the mean value of number of leaves in all AMF inoculated buckets were higher than the Control except at 4WAP where the Control recorded the highest mean value of 4.75. At 4WAP, the Control recorded the highest value (4.75) while Gigaspora specie recorded the least value (4.17), although no statistical difference was recorded among treatments. At 8WAP, Gigaspora specie recorded the highest mean value (6.25) while the Control (5.62) recorded the least value. The G.clarium and G.mossea recorded (6.1) and (6.0) respectively, but no statistical difference was recorded among treatments. At 10WAP, G.mossea recorded the highest value (8.62) while the Control recorded the least value (7.50). The G.mossea is statistically higher than the Control (7.50) but the same as Gigaspora specie (7.75) and G.clarium (7.88), respectively.
Although, the mean values of stem girth of maize plants inoculated with AMF were higher than the Control except at 10WAP but there was no significant difference was recorded among treatment through the experimental period. At 4WAP, G.clarium recorded the highest value (1.500 cm) while Gigaspora specie recorded the least value (1.413 cm). No statistical difference between treatments. At 8 WAP, G.mossea recorded the highest value (4.330 cm) while Gigaspora specie recorded the least value (3.710 cm). However, no statistical difference was recorded among treatments. At 10WAP, the Control recorded the highest value (4.500 cm) while G.clarium recorded the least value (4.260 cm). Although G.mossea recorded (4.313 cm) and Gigaspora specie recorded (4.34 cm), there is no statistical difference among treatments.
Table 1: Effect of Mycorrhizal fungi on selected growth parameters of maize planted on sewage contaminated soil.
TRT= Treatment, PH = Plant Height, SG = Stem Girth, LA = Leaf Area, NL = Number of leaves, G.sp = Gigaspora sp, G.c = Glomus clarium, G.m = Glomus mossae, WAP = week after planting.
PHYSIOCHEMICAL PROPERTIES OF THE SOIL
Table 2 shows the physical and chemical properties of the analysed soil.
The SBSA (85.20 %) and the Control (85.20 %) respectively recorded the highest sand content while Gigaspora specie recorded the least value (85.0 %). It is observed that SBSA and the Control are statistically higher than all AMF inoculated soil. It is worthy to note that, there is no significant difference between SBSA and the Control. However buckets inoculated with G.clarium (85.10 %) is statistically higher in sand content than buckets inoculated with Gigaspora specie (85.00 %) followed by buckets inoculated with G.mossea (84.50 %).
The bucket inoculated with G.mossea (4.50 %) is statistically highest among the treatments. It is observed that bucket inoculated with G.mossea is statistically higher than that of G.clarium (4.06 %) and Gigaspora specie (3.90 %) followed by SBSA (2.90 %) and Control (2.85 %). It is worthy to note that there is no significant difference between G.clarium and Gigaspora specie likewise SBSA and the Control.
The Control has the highest clay content with mean value (11.95 %) while G.clarium has the least clay content with mean value (10.90 %). Control and SBSA (11.90 %) are statistically the same but higher than Gigaspora specie (11.10 %) and G.mossea (11.00 %) Soil samples inoculated with Gigaspora specie and G.mossea are statistically the same but higher than G.clarium (10.90 %).
Table 2: Physico-chemical Properties of the analysed Soil
SBSA = Soil Before Sewage Application, means with the same alphabets show no statistical difference while different alphabets show statistical difference, M0 = Control, M1 = Gigaspora sp, M2 = Glomus clarium , M3 = Glomus mossea. Trt = Treatment
The results show that G.clarium recorded the highest pH mean value (5.22 %) while Soil before Sewage Application (SBSA) recorded the least pH mean value (4.96 %). G.clarium (5.22 %), G.mossea (5.21 %) and, Gigaspora specie (5.21 %) show no significant difference among the treatments but are significantly higher than the Control (5.00 %) and SBSA (4.96 %) respectively. However, the Control and SBSA are statistically the same.
Total Organic Carbon (TOC)
The TOC of G.mossea inoculated soil has the highest mean value (0.490 %) while SBSA had the least value (0.20 %). Soil inoculated with G.mossea (0.49 %) is statistically higher than G.clarium (0.38 %). The Control (0.30 %) is statistically the same with Gigaspora specie (0.32 %) but significantly higher than SBSA (0.2 %).
Total Nitrogen (TN)
The soil inoculated with G.mossea (0.45 %) has the highest TN content while SBSA has the least value (0.17 %). The soil inoculated with G.mossea is statistically higher than that of G.clarium (0.32 %) followed by Gigaspora specie, The Control and SBSA respectively. There is no statistical difference between the Control and SBSA. The Control and plant inoculated with Gigaspora specie are significantly the same.
Available phosphorus (Av.P)
Both SBSA and Control recorded the least Av.P value (1.36 mg/kg) respectively while G.mossea recorded the highest Av. P value (5.42 mg/kg). G.mossea is statistically higher than G.clarium (4.15 mg/kg) and Gigaspora specie (3.42 mg/kg), while Gigaspora specie is statistically lower than G.Clarium.
The highest calcium content was recorded in soil inoculated with G.mossea (2.80 Cmol/kg) while the least was the Control (2.20 Cmol/kg). The G.mossea is statistical higher than other treatments, although G.clarium (2.60 Cmol/kg) and Gigaspora specie (2.55 Cmol/kg) are significantly the same, they are statistically higher than Control and SBSA. The SBSA and Control are statistically the same.
Soil inoculated with G.mossea has the highest value (2.80 Cmol/kg) of Mg from the results shown in Table 5 while the Control has the least value (2.06 Cmol/kg).The soil inoculated with G.mossea is statistically higher than soil inoculated with G.clarium (2.45 Cmol/kg), followed by Gigaspora specie (2.40 Cmol/kg), then SBSA (2.17 Cmol/kg) and lastly the Control (2.06 Cmol/kg). There is no statistical difference between G.clarium and Gigaspora specie.
Soil inoculated with G.mossea has the highest value (0.18 Cmol/kg) while SBSA and Control have the least value (0.12 Cmol/kg) respectively. The soil inoculated with G.mossea is statistically higher than G.clarium (0.15 Cmol/kg), followed by Gigaspora specie (0.13 Cmol/kg), then SBSA and the Control (2.06 Cmol/kg). There is no statistical difference between Gigaspora specie, Control and SBSA.
Soil inoculated with G.mossea has the highest value (0.19 Cmol/kg) while SBSA has the least value (0.14 Cmol/kg). G.mossea is statistically higher than G.clarium (0.15 Cmol/kg), Gigaspora specie (0.15 Cmol/kg), Control (0.15 Cmol/kg), SBSA (0.14 Cmol/kg). There is no statistical difference between G.clarium, Gigaspora specie, the Control and SBSA.
Cation Exchange Capacity (CEC)
Soil inoculated with G.mossea has the highest CEC value (5.97 Cmol/kg) while the Control has the least value (4.53 Cmol/kg). Soil inoculated with G.mossea is statistically higher than G.clarium (5.35 Cmol/kg), followed by Gigaspora specie (5.23 Cmol/kg), then SBSA (4.66 Cmol/kg) and lastly Control (4.53 Cmol/kg). There is no statistical difference between G.clarium and Gigaspora specie and also between the Control and SBSA.
Chemical properties of Selected Trace elements
The chemical properties of selected trace elements are shown in Table 3 below.
Table 3: Chemical properties of Selected Trace Elements on the analyzed soil Samples
SBSA = Soil Before Sewage Application, means with the same alphabets show no statistical difference while different alphabets show statistical difference, M0 = Control, M1 = Gigaspora sp, M2 = Glomus clarium , M3 = Glomus mossea, Trt = treatment
The soil sample before sewage application (SBSA) recorded Copper mean value of 1.4 mg/kg before planting but statistically increased with the application of sewage in Control to value 4.0 mg/kg. When AMF was added, soil inoculated with G.mossea recorded the least value 1.7 mg/kg but not statistically difference with SBSA. Soil inoculated with G.clarium recorded a mean Cu value of 2.7 mg/kg which is statistical lower than soil inoculated with Gigaspora specie (3.6 mg/kg) but higher than SBSA and soil inoculated with G.mossea.
The SBSA recorded Zn mean value of 7.0 mg/kg but increased to 10.0 mg/kg in the Control. Soil inoculated with G.mossea (5.5 mg/kg) recorded the least mean value of Zinc followed by G.clarium (7.0 mg/kg) and lastly Gigaspora specie (9.0 mg/kg). The G.mossea is statistical lower than other treatments. The G.clarium and SBSA are statistically the same but lower than Gigaspora specie and the Control. However, Gigaspora specie shows no significant difference with the Control
The SBSA recorded lead mean value of 2.0 mg/kg which is statistically the same with soil inoculated with G.clarium (2.0 mg/kg) and G.mossea (1.8 mg/kg) respectively, but they are significantly lower than soil inoculated with Gigaspora specie and Control. However, Soil inoculated with Gigaspora specie (4.0 mg/kg) is statistically the same with Control.
The Control has the highest Nickel value 0.9 mg/kg while SBSA has the least value 0.02 mg/kg. The SBSA is statistically lower than Soil inoculated with G.mossea (0.03 mg/kg) which is statistically lower than soil inoculated with G.clarium (0.06 mg/kg) and Gigaspora specie (0.06 mg/kg) respectively and they are all statistically lower than Control. The soil inoculated with G.clarium and Gigaspora specie are not significantly different from each other.
Generally the value for Sand content statistically decreased in AMF inoculated samples (84.5 %) than in SBSA (85.2 %) and Control (85.2 %). Similarly, Clay content statistically decreased in AMF inoculated (10.9 %) than its value in BP (11.9 %) and in Control (11.95 %). On the other hand, the Silt content increased in AMF inoculated samples (4.5 %) compared to those of the Control (2.85 %) and SBP (2.9 %). It is obvious that AMF inoculation positively influenced soil aggregation in the study. This observation is in agreement with the studies of Tisdall and Oades (1982) who reported that AMF and other fungi are hypothesized to be important for soil aggregation at the macro-aggregate level, where direct hyphal involvement is thought to be most pronounced. Additionally, as micro-aggregates are thought to form most frequently within macro-aggregates, AMF-facilitated stabilization of macro-aggregates would be expected to result indirectly in micro-aggregate formation. Also, AMF and their diversity have been shown to be important controllers of the productivity of plant communities (van der Heijden et al., 1998), in part via their effects on plant community composition.
The pH in AMF inoculated soils ranges between (5.00 – 5. 22) which slightly deviated from the findings of Wang et al. (1993) who reported that AMF develop optimum number of spores at pH value ranging from (5.5 – 7.5). The increase in soil pH and reduction in amount of trace elements as shown in Table 2 and 3 respectively, supports earlier report by Shen et al., (2006) who observed reduction in available heavy metals as a result of AMF inoculation due to significant increase in soil pH and strongly supports the use of AMF to curb the effect of sewage contamination in soil.
The TOC is the remains of plant and animals of microorganisms at various stages of decomposition. The result obtained from the analysis, shows significant increase in the TOC of soil inoculated with G.mossea (0.49 mg/kg) when compared with the Control (0.30 mg/kg) and SBSA (0.20 mg/kg). This observation is in support of earlier findings by Cardoso and Kuyper (2006) who reported that AMF plays role in the carbon cycling thus; increasing AMF will increase carbon flow into soil.
There was an increase in P content of AMF inoculated soils, G.mossea (5.42 mg/kg) compared with SBSA (1.36 mg/kg) and Control values (1.36 mg/kg) as shown in Table 2. This is in tandem with the findings of (Smith et al., 2003); (Buckling and Shacker-Hill, 2005) who reported that Phosphorus is made available in the soil due to the presence of mycorrhiza. The increase in the agronomic parameters of plants as recorded for Plant height of G.mossea with mean value of 32.0 cm at 4WAP and 92.5 cm at 8WAP for AMF inoculated soils could be as a result of sufficient P availability unlike the plants in the Control thus support earlier findings by Bucher (2007) who reported that under stressed condition, AMF improves phosphorus supply in the soil.
The result obtained shows an increase in Nitrogen when soil inoculated with AMF (0.45 mg/kg) is compared with the Control value (0.20 mg/kg) and SBP (0.17 mg/kg) as shown in Table 2. The same trend is observed in growth parameters at 8WAP where G.mossea recorded the significantly highest plant height mean value (92.50 cm) and stem girth mean value (4.33 cm) compared with the Control with least plant mean value (81.5 cm) and (3.73 cm) for plant height and stem girth respectively. These results confirm earlier report of Hawkins et al., (2000) which states that AMF enhances Nitrogen uptake and utilization in soil.
The soil inoculated with AMF had the significantly highest K value (0.18 Cmol/kg), Ca value (2.80 Cmol/kg), Mg value (2.80 Cmol/kg) and CEC value (5.97 Cmol/kg) when compared to their SBSA value, K value (0.12 Cmol/kg), Ca value (2.23 Cmol/kg), Mg value (2.17 Cmol/kg) and CEC value (4.66 Cmol/kg). This is agreement with the report of Sharifi et al., (2007); Terrer et al.,(2016) and Turrini et al., (2017) which states that the main function of AMF is to enhance the nutrient uptake of elements like K, Ca, Mg and CEC by host plants, improves the nutrient metabolism capacity and nutrient level and promote plant growth. In addition, the observation also in agreement with the findings of Nazeri et al., (2013) and Leigh et al., (2009) who reported that when soil is inoculated with AMF, there is an increase the uptake of nutrients in the soil, such as Calcuim and Magnesium.
The amount of trace elements as shown in the result is higher in SBSA when compared with soil inoculated with AMF; this could be traceable to the presence of these metallic elements in the sewage applied. This observation supports the finding of Tyla, et al., (2016) who reported that Sewage contains heavy metals such as Cadium (Cd), Zinc (Zn), Lead (Pb), Copper (Cu) etc. The metallic elements in inoculated soil samples significantly reduced, this is due to the presence of AMF in the soil as it has ability to survive in contaminated soils. The mean value of Cu in soil inoculated with G.mossea statistically reduced when compared with SBSA and this observation agrees with the findings of Lee and George (2005) who reported that G.mossea contributes to 75 % of Cu uptake in contaminated soil.
In this study, AMF significantly increased the growth rate of maize in sewage contaminated soil, as seen in the Leaf Area recorded at 8WAP where plants inoculated with AMF G.Mossea (320 cm2) is significantly higher than the Control with mean value (187 cm2) and this is in agreement with Shen et al,. (2006) who observed that AMF inoculation on maize enhances plant growth, P content and plant tolerance to metals like Zn either by lowering its level in the uptake process or through up-taking it into the extra-radical mycelium of the AMF and also the works of (Kaldorf et al., 1999; Aroca et al., 2013; Wu et al., 2014; Hashem et al., 2016) who reported that AMF has the ability to enhance plants capacity to uptake essential nutrient while partially excluding non-essential ones. Thus AMF inoculation can be considered a feasible practical method in reclaiming soils contaminated by sewage (Turnau et al., 2006).
This report reveals that sewage contaminated soil is deleterious in maize cultivation. AMF inoculation ameliorates the deleterious effect of sewage in agricultural soils as it aids the absorption and utilization of nutrient elements like phosphorus.
Comparatively, assessing the three (3) species of Mycorrhiza used during the experiment; Glomus mossea outperformed G.clarium and Gigaspora specie both in plant and soil parameters. Therefore, Glomus mossea is recommended as Soil amendment and biofertilizer for maize cultivation in soil contaminated with sewage. Furthermore, extensive research is needed in understanding the biochemical dynamics of Mycorrhiza and trace metals found in sewage contaminated soils.
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