Open Access is an initiative that aims to make scientific research freely available to all. To date our community has made over 100 million downloads. It’s based on principles of collaboration, unobstructed discovery, and, most importantly, scientific progression. As PhD students, we found it difficult to access the research we needed, so we decided to create a new Open Access publisher that levels the playing field for scientists across the world. How? By making research easy to access, and puts the academic needs of the researchers before the business interests of publishers.
We are a community of more than 103,000 authors and editors from 3,291 institutions spanning 160 countries, including Nobel Prize winners and some of the world’s most-cited researchers. Publishing on IntechOpen allows authors to earn citations and find new collaborators, meaning more people see your work not only from your own field of study, but from other related fields too.
To purchase hard copies of this book, please contact the representative in India:
CBS Publishers & Distributors Pvt. Ltd.
www.cbspd.com
|
customercare@cbspd.com
The research proposes a biofertiliser from mycorrhiza and rhizobium evaluating its antagonistic capacity and biotisation in the cultivation of vegetables with a DCA, the sample considers the potato, pea, and barley in the Huasahuasi Peruvian District, with nine treatments in three formulas, considering a control group without inoculation and two repetitions. As a result, the optimal formula is obtained with 300 g of mycorrhizal and rhizobium strains + 500 g of black soil + 200 g of potato peel crust, which has an effective antagonistic capacity of 100% in pea cultivation, 90% in the barley, and 85% in the potato, besides that it achieves a biotisation in the cultivation of peas of 95%, in the barley 100% and in the potato 90%.
Latin American soils have low yields, chemical fertilisers are expensive, there are phytosanitary problems, soil deterioration and nitrogen deficiency in agricultural land, and at the international level, there is a search for ways to stop soil erosion, which is of great importance for the biological diversity of vegetation and fauna (Figure 1) [1].
Figure 1.
Soil problems.
One emblematic case is camu camu, which decreased to 1.5–2.5 t/ha during 2017 due to the reduction of N, K, and Mg in the soil. For this reason, it was proposed to increase production levels with biofertilizers by using cow manure, chicken manure, island guano, and river sediment.
The use of biofertilisers promotes insect repellency, increases resistance to pest and pathogen attacks through their odour (Figure 2) [2].
Figure 2.
Benefit of biofertiliser.
Climate change challenges agriculture, and variations in production and costs directly affect farmers [3]. In other countries, there are no soil quality problems, but pesticide residuals in products, such as tomatoes and cape gooseberries, with up to 10 pesticides found in the fruit and on the skin in concentrations of more than 0.002 ppm, toxic compounds, such as sulfotep, phorate, heptachlor, aldrin, endosulfan sulphate and I, making export impossible due to the minimum sanitary quality requirements [4].
In response, work is being done to raise awareness, proposing other forms of energy use such as alternative energies [5] and the use of arbuscular mycorrhizal fungi (AMF) together with Twin-N as biofertilisers in potato cultivation, which would completely replace the use of chemical fertilisers with a yield per hectare of more than 116% compared to traditional fertilisation and a mostly healthy harvest of tubers with 1–10% skin lesions [6], moulds and beneficial bacteria to induce nodulation, inhibit the development of pathogenic microorganisms, fix nitrogen and other nutrients in plants, has been studied as an option for potential impact.
Mycorrhizae cover 95% of the requirements in the production of walnuts, being the production needs of 30% of nitrogen and 50% of potassium and phosphorus, the costs were 40.8% of the income from sales [7], there are studies whereby providing rhizobia a good quantity of nodules was obtained with very low weights with respect to the optimum values, but this is remedied after inoculating B. japonicum and Nod factors, offering a biotechnological alternative of acceptable yield [8].
But the antagonistic activity is another important factor, a study measures Trichoderma harzianum strains against Rhizotonia spp., Nakatea sigmoide, and Sclerotum folfsii, making T. harzianum superior in antagonism and antiparasitic activity against Garrido [9].
Work was carried out on wheat grains, obtaining an increase in Nitrogen (2 to 15 N/ha) and dry matter absorbed of 20–40% of that applied biofertilisers improve nutrient absorption [10].
arbuscular mycorrhizal fungi, hydrogen sulphate, and Mucoromycotina fungi are studied, which colonise 78.1% of the species, of which only 56.2% are considered to be mycorrhizal [11].
When inoculating native rhizobia on peas (Pisum sativum), 40% of the crops show the formation of nodules in symbiosis, but only 10% show their efficiency in terms of nodulation percentage and speed (Figure 3) [12].
Figure 3.
Rhizobium action.
Organic fertilisers in sunflower give the highest availability of nutrients, improve the weather and conditions suitable for this crop, increase the achene protein (APC), and highlight the need for water supply and sunshine on the performance of the plant development. The benefits of biofertilisation are an increase in available N which increases soil microbial activity, increases P and K content, dry matter and protein yield, the biofertiliser that obtained the highest rates of 48% oil and 14% protein is goat manure [13].
Biofertilisers were found to increase P, Ca, and Mg values but were not very effective in coffee plantations, well conventional planting systems had no differences with respect to plagiotropic branches as well as fertiliser application and type of planting [14]. Similarly, with the addition of BMV-biofertiliser, the increase in N fell and the contents of Cu and Fe decreased linearly with the increase in biofertiliser. The loss of volatile N is indicated by the alkalinity and aggregation of Ca and Mg in the oil [15].
A biofertiliser obtained by anaerobic digestion of cassava effluent was applied to the development of Crambe plants, the results indicated that the higher the percentage of biofertiliser, the oil values obtained were lower than those of the control, even that the minimum value was achieved with the highest inoculation, in addition to potassium deficiency results in decreased productivity in Crambe grains [16]. Another case is the application of cattle manure biofertiliser to strawberry plants, where it was found that production was greater than 1,250 ml/plant/week in a protected environment and sprayed with cold water and white soil, obtaining the largest fruit size in diameter and length, but with less soluble solids content (Brix) than those grown in the environment in full sun [17].
Adding saltwater to soybean reduces photosynthesis, stomatal conductance and transpiration, with low intensity when inoculated with aerobically fermented bovine biofertiliser [18], demonstrating a plant protection mechanism. When evaluating the ectomycorrhizal fungus of pine, the accumulation of heavy metals in the roots of plants with ectomycorrhizal fungi was noted, which, contrary to expectations, had fewer shoots with this type of fungi, there was no difference with the control with respect to the rhizosphere, but there was a predominance of acidobacteria, actinobacteria, and proteobacteria [19].
This study analyses mycorrhiza strains isolated from pine fungus and rhizobium isolated from pea root, thus promoting their use as biofertiliser and taking advantage of their antagonistic capacity, considering their biotisation generated from these microorganisms in plants.
It is estimated that this process contributes between 60 and 80% of biological nitrogen fixation and this symbiosis provides a considerable part of combined nutrients and nitrogen in the soil and allows plants to grow without synthetic fertilisers and without impoverishing soils [20].
The present research work was carried out in the district of San Juan de Huasahuasi, located 48 km from the district of Tarma, at an altitude of 2,751 m above sea level. The raw material consists of mycorrhiza and rhizobium isolated from pea root and pine fungus, rhizobium is diazotrophic bacteria that have the ability to fix N in plant nodules and mycorrhizae are the double absorption organs that are formed when symbiont fungi live inside healthy absorption organs (roots, rhizomes, or stems) [7, 12, 21]. Black soil and potato peel bran were used as inputs, oil and potato dextrose agar were also used, the equipment used was a microorganism incubator and an analytical balance (Figure 4) [22, 23, 24, 25, 26].
Figure 4.
Elements of biofertiliser.
Vegetable crops were sampled—potatoes, peas, and barley in the Huasahuasi district of Tarma province.
Among the methods used were the Association of Analytical Communities in vitro sowing method and colony counting. Once the product was obtained, a physical-chemical characterisation was carried out, evaluating fertility, antagonistic capacity, and biotisation (Figure 5).
Figure 5.
Characterisation of the biofertiliser.
The experimental process consists of obtaining strains of microorganisms and the biofertiliser mycorrhiza and rhizobium from pea root and pine fungus through two stages:
The first stage consists of obtaining strains of mycorrhiza and rhizobium microorganisms, obtained from pea root and pine fungus, which is described in Figure 6.
Figure 6.
First stage: breeding of strains.
The second stage consists of obtaining the optimal biofertiliser formulation of mycorrhiza and rhizobium, which is shown in Figures 7 and 8.
Figure 7.
Second stage: obtaining the optimal biofertiliser formula.
Figure 8.
Inoculation of biofertiliser formulation to pea seed.
The statistical evaluation assessed the percentage effect of the biofertiliser in relation to its antagonistic capacity and biotisation, applying a completely randomised design (CRD) with the factorial arrangement and two replications [22], the factors were as follows:
Factor A: Inoculation of mycorrhiza and rhizobium biofertiliser formulation (F1, F2, and F3).
Factor B: Vegetable crops (potato, pea, and barley).
With a factorial arrangement of 3A × 3B = nine treatments and two replicates, the antagonistic and biotic effect of mycorrhiza and rhizobium fertiliser on vegetable crops (potato, pea and barley) is compared (Table 1).
Repetitions
Treatments
Factor A
1
2
3
1
Factor B
1
F1C1
F2C1
F3C1
2
F1C2
F2C2
F3C2
3
F1C3
F2C3
F3C3
2
Factor B
1
F1C1
F2C1
F3C1
2
F1C2
F2C2
F3C2
3
F1C3
F2C3
F3C3
Table 1.
Relationship of biofertiliser formulation and plant cultivation.
F = Biofertiliser formula; F1 = 100 g of strains of microorganisms + 500 g of black soil and 200 g of potato peel bran; F2 = 200 g of strains of microorganisms + 500 g of black soil and 200 g of potato peel grist; F3 = 300 g of strains of microorganisms + 500 g of black soil + 200 g of potato peel bran; C = Vegetable cultivation (potato, pea, and barley); C1 = Potato; C2 = Peas; and C3 = Barley.
The analysis was applied to the treatments of the biofertiliser of mycorrhiza and rhizobium inoculating the sample with 50 g for 200 g of seed, an effective antagonistic capacity of 100% was obtained in the pea crop, 90% in barley and 85% in potato, of the biofertiliser of mycorrhiza and rhizobium with the formula F3, the biofertiliser with the formula F2 inoculated on the pea crop obtained a result of 85% effectiveness, 80% on barley, and 65% on the potato crop, in comparison to these two formulas where the result was effective, the biofertiliser with the formula F1 the results were not very significant as the percentage of effectiveness on the pea crop was 45%, barley 45%, and potato 35%. The optimal formula of the biofertiliser of mycorrhiza and rhizobium was F3, to be used in the cultivation of vegetables, obtaining a significant result in the antagonistic capacity of the biofertiliser (Table 2).
Results
Crops
Formula
C1 Pea (%)
C2 Barley (%)
C3 Potato (%)
Effectiveness
F1
45
45
35
No
F2
85
80
65
Effective
F3
100
90
85
Effective
Table 2.
Comparison of the effectiveness of antagonistic capacity.
K = Ratio of strains of microorganisms 500 g mycorrhiza (pine fungus) + 500 g rhizobium (pea root).
3.2 Analysis of biotisation in vegetable crops
This analysis was carried out to verify the growth of the root system, the acclimatisation phase, and the increase in the functionality of the roots and, consequently, the nutritional and water status of the vegetable crops. The results obtained in relation to the inoculated formula and the vegetable crop used as a sample, which in this case was potato, pea, and barley, are shown in Table 3.
Results
Crops
Formula
C1 Pea (%)
C2 Barley (%)
C3 Potato (%)
Effectiveness
F1
55
60
35
No
F2
95
95
90
Effective
F3
95
100
90
Effective
Table 3.
Comparison of the effectiveness of biotisation.
According to the results shown in Table 3, we can determine that the mycorrhiza and rhizobium biofertiliser has an effective effect on the biotisation of the plant crop by increasing the number of strains of microorganisms in the biofertiliser formula.
3.3 Balance of matter of the obtaining of the biofertiliser of Mycorrhiza and Rhizobium according to the optimal formula.
To determine the yield of the optimum formula of the mycorrhiza and Rhizobium biofertiliser, a balance of matter was carried out starting with 10 kg of pea root and 10 kg of pine fungus, the roots and fungi were selected taking into account the optimum characteristics, 16.7% was lost. The conditioning operation discards the unusable parts, stems and filaments, losing 13% during the drying operation and eliminating 55% of the water. The dry material is milled with a loss of 2%, thus obtaining a yield of 13.3 % with respect to the initial raw material (1,333 kg).
1,333 kg (50% dry pea root + 50% dry pine fungus) is mixed with 2,221 kg of black soil 0,888 kg of potato peel flour, making a total of 4,442 kg of biofertiliser for every 10 kg of fresh pea root and 10 kg of fresh pine fungus.
The economic feasibility assessment was carried out in each production phase and two stages.
The first stage is called obtaining the strains from the pea root and the pine fungus, both of the same proportion, these are selected, cut, crushed, and dried, in each process, there are diligently measured losses between them; there is a loss of 86.7%. Based on these losses, it can be deduced that the yield in this first stage amounts to 13.3% (Table 4).
First phase
%
Kg
Pea root Kg
10
Pine mushroom Kg
10
Selection
Loss 16.7
3.34
Shredded
Loss 13
2.6
Crushed
Loss 2
0.4
Dried
Loss 55
11
Result
Yield 13.3
2.66
Table 4.
Result first phase.
The second phase deals with the elaboration of the biofertiliser product from the previously obtained strains. This consists of a mixture of strains from the first phase (30%), black soil (50%), and potato peel flour (20%) (Tables 5 and 6).
Product
Quantity required Kg
Cost × 1 Kg
Total
Pea root Kg
10
2.00
20.00
Pine mushroom Kg
10
14.00
140.00
160.00
Table 5.
Cost first phase.
Quantity (Kg)
Cost × 1 Kg
Cost PEN
Cost GBP
First phase strains (30%)
2,666
160.00
29.79
Black soil (50%)
4,442
1.50
6,663
1.24
potato peel flour (20%)
1,776
1.00
1,776
0.33
Subtotal
8,884
Labour
3 wages
30.00
90.00
16.76
Cost 8,884 Kg
258,439
48.13
Cost Kg
29.09
5.82
Table 6.
Cost second phase.
In the beginning, 10 kg of pea root at a price of 2.00 Peruvian suns (PEN) and 10 kg of pine fungus at 14.00 PEN each Kg are used, making expenses of 20.00 and 140.00, respectively.
For the second phase, black soil is required in quantities of 4,442 kg equivalent to 50% of the total mixture of 1.5 soles, amounting to 6,663 soles and potato peel flour in quantities of 1,776 kg equivalent to 20% of 1.00 PEN, amounting to 1,776 PEN.
The labour required is three daily wages of 30.00 each making 90.00 PEN.
With these costs and wastage, a total of 8,884 kg costs 258,439 PEN, which is 29.09 soles or 5.82 GBP per kg of biofertiliser.
The yield tests in the field show a yield of 60% when using 30 g per kg of seed potato, however, it is necessary to carry out further field tests to prove the effectiveness of each product.
Regarding the analysis of the competition, there are products, such as Trichoderma, which in its presentation of 100 g has a cost of 4.00 GBP, shipped in Ecuador in South America. Another product is the blood meal whose price per 1 Kg is 140 GBP on average and the Mycoracine that in the presentation of 500 g has a value of 543.50 GBP or the Bacillus Subtilis of 500 g at a cost of 445.00 GBP
Antagonism is the direct inhibitory activity exerted by one microorganism on another and controls it biologically by attenuating damage to growth systems [24].
The antagonism test against soil phytopathogens was measured with respect to the fungus Fusarium Solani, the F2 and F3 formulations were effective in the antagonistic capacity against this fungus, which represents 100% of the strains, This could be due to the fact that the number of Streptomyces strains evaluated exceeded 4,000, since the inhibition zones obtained are equivalent to the average inhibition percentages obtained in the present study [23], the mechanism of antibiosis effect is presumed to be by means of inhibitory metabolites, in the same way as [24] or by repellency, as in Abanto-Rodríguez et al. [2].
Regarding the recovery of soils, the product obtained is easily used in the prevention of soil erosion [1] and can be considered as a new form of chemical energy alternative to conventional ones [5], as both mycorrhiza- and rhizobium-based products increase the amount of N in the soil as well as K and Mg, the results are similar to those obtained by Borges et al. [13], Figueiredo et al. [14], Cardoso et al. [15], and de Sousa et al. [18] for their effectiveness in biotisation.
The methodology for obtaining the biofertiliser differs from those obtained in aerobic digestion [16], in that the method proposed in this study allows the creation of a product directly proportional to its amount of addition.
Biotisation is the use of fungi and bacteria on plants that achieve acclimatisation and creation of beneficial rhizosphere [25], the effect of biotisation showed the growth of the root system, its acclimatisation and increased root functionality, comparisons show that F3 in barley is 100% effective in direct benefit to farmers [3] and similar to the findings of Flore-Córdova with the ability to replace traditional fertilisers [7] even at a lower cost as it is not necessary to inoculate boosters like Fornasero and Toniutti [8] to achieve results, These results are supported by Grageda-Cabrera et al. [10] and Lara-pérez et al. [11], but they are better than Moreno-chirinos et al. [12] as they achieve nodule formation higher than 40 % of crops, with values of 95 % in peas and even the less effective F1 formula shows 55%.
The biofertiliser based on mycorrhiza and rhizobium is antagonistic to the fungus Fusarium Solani, increasing its antagonistic activity by increasing the dose of these strains in the formulation.
The biofertiliser based on mycorrhiza and rhizobium is able to recover soils for the cultivation of peas, potatoes and barley.
The biofertilisation effect of the biofertiliser understudy is higher in barley (100% effective) due to its higher capacity to produce substances that stimulate plant growth.
1.FAO. Día mundial del suelo. 2019. Available from: http://www.fao.org/world-soil-day/es/ [Accessed: October 11, 2019]
2.Abanto-Rodríguez C, Mori GMS, Panduro MHP, Castro EVV, Dávila EJP, de Oliveira EM. Uso de biofertilizantes en el desarrollo vegetativo y productivo de plantas de camu-camu en Ucayali, Perú. Revista Ceres. 2019;66(2):108-116
3.Álvaro Hernán Alarcón JDS, Arias G, Díaz CJ. Sistema de control automático de variables climáticas para optimizar el rendimiento de cultivos bajo cubierta. Ingeniería Solidaria. 2017;2017:1-17
4.Ávila-Orozco FD, León-Gallón LM, Pinzón-Fandiño MI, Londoño-Orozco A, Gutiérrez-Cifuentes JA. Residualidad de fitosanitarios en tomate y uchuva cultivados en Quindío (Colombia). Corpoica Ciencia y Tecnologia Agropecuaria. 2017;18(3):571-582
5.Gonzáles Y. Acciones preventivas del deterioro de los suelos como alternativa para fomentar la cultura conservacionista. Revista Science. 2019;53(9):1689-1699
6.Castillo C, Huenchuleo MJ, Michaud A, Solano J. Micorrización en un cultivo de papa adicionado del biofertilizante Twin-N establecido en un Andisol de la Región de La Araucanía. Idesia. 2016;34(1):39-45
7.Flores-cordova MA, Parra JMS, Piña FJ, Pérez R, Sánchez E. Contribution of nutrients, organic amendments and mycorrhizae on the yield components in pecan walnut ( Carya ilinoinensis). Cultivos Tropicales. 2018;39(1):35-42
8.Fornasero LV, Toniutti MA. Evaluación de la nodulación y rendimiento del cultivo de soja con la aplicación de distintas formulaciones de inoculantes. Fave—Sección ciencias agrarias. 2015;14(1):79-90
9.Garrido M. Capacidad antagónica de Trichoderma harzianum frente a Rhizoctonia, Nakatea sigmoidea y Sclerotium rolfsii y su efecto en cepas nativas de Trichoderma aisladas de cultivos de arroz. Scientia Agropecuaria. 2019;10(2):199-206
10.Grageda-Cabrera OA, González-Figueroa SS, Vera-Nuñez JA, Aguirre-Medina JF, Peña-Cabriales JJ. Efecto de los biofertilizantes sobre la asimilación de nitrógeno por el cultivo de trigo. Revista mexicana de ciencias agrícolas. 2018;9(2):281-289
11.Lara-pérez LA, Zulueta-rodríguez R, Andrade-torres A. Micorriza arbuscular, Mucoromycotina y hongos septados oscuros en helechos y licófitas con distribución en México : una revisión global. Revista de Biología Tropical. 2017;65(101):1062-1081
12.Moreno-chirinos ZE, Valdez-núñez RA, Soriano BS, Ruesta-campoverde NA. Eficiencia en la nodulación por rizobios nativos, procedentes de nódulos de Pisum sativum ‘arveja’ colectados de diferentes departamentos del Perú. Scientia Agropecuaria. 2016;7(3):165-172
13.Borges FRM, Bezerra FML, Marinho AB, Ramos EG, Adriano JDNJ. Goat manure fertilization and irrigation on production components of sunflower. Revista Caatinga. 2019;32(1):211-221
14.Figueiredo LH, Miranda GRB, Vilella PMF. Biofertilizer use associated with different forms of planting in the development of the initial coffee arabic. Coffee Science. 2017;12(4):463-470
15.Cardoso MO, Berni RF, Antonio IC, Kano C. Growth, production and nutrients in coriander cultivated with biofertilizer. Horticultura Brasileira. 2017;35(4):583-590
16.Neves AC, Bergamini CN, de Leonardo RO, Gonçalves MP, Zenatti DC, Hermes E. Effect of biofertilizer obtained by anaerobic digestion of cassava effluent on the development of crambe plants. Revista Brasileira de Engenharia Agrícola e Ambiental. 2017;21(10):681-685
17.Dos Santos EM, Viana TVDA, De Sousa GG, De Azevedo BM, Moraes JGL. Yield and quality of strawberry fruits fertilized with bovine biofertilizer. Revista Caatinga. 2019;32(1):16-26
18.de Sousa GG, dos Rodrigues VS, da Soares SC, Damasceno ÍN, Fiusa JN, Saraiva SEL. Irrigation with saline water in soybean (Glycine max (L.) Merr.) in a soil with bovine biofertilizer. Revista Brasileira de Engenharia Agrícola e Ambiental. 2018;22(9):604-609
19.Yu P, Sun Y, Huang Z, Zhu F, Sun Y, Jiang L. The effects of ectomycorrhizal fungi on heavy metals’ transport in Pinus massoniana and bacteria community in rhizosphere soil in mine tailing area. Journal of Hazardous Materials. 2020;381(July):121203
20.Ventura S, Bernilla BS. Efecto de Trichoderma viride y Bradyrhizobium yuanmingense en el crecimiento de Capsicum Annuum en condiciones de laboratorio. Rebiolest. 2014;2(2)
21.Ramírez M, Rolon M, Moncada A, Serralde DP. Biofertilización con hongos formadores de micorrizas arbusculares (HFMA) en especies forestales en vivero. Biotecnología En El Sector Agropecuario Y Agroindustrial;16(2):15-25
22.Sampieri RH, Collado CF, Lucio PB. Metodología de la investigación. 5ta ed. México D.F.: McGraw-Hill / Interamericana Editores, S A de C V; 2010
23.Ikeda K, Saito H, Sato SI, Kodama T. Growth-inhibition effect of Streptomyces sp. 39L40C to mulberry twig blight pathogen, Fusarium lateritium f. sp. mori. Journal of Sericultural Science of Japan. 2000;69(3):163-168
24.Orihuela M. Morfotipos de hongos endomicorrizógenos asociado al maíz amiláceo (Zea mays L. amiláceo), y su capacidad antagónica para el control de Fusarium oxysporum. Universidad Nacional de San Cristobal de Huamanga. 2016;1(1):7-120
25.Infante D, Martinez B, Gonzáles N, Reyes Y. Mecanismos de acción de Trichoderma frente a hongos fitopatogenos. Protección. 2009;24(2):14-21
26.Carrazana D, Santos A, Alderete Y, Galvez D, Cupull R, Navarro M. Bacteria endófita latente no vitropatógena en el cultivo in vitro de Xanthosoma sagittifolium (L. Schot). Centro Agrícola. 2011;38(4):21-29
Written By
Henry Juan Javier Ninahuaman and Grimaldo Quispe Santivañez
Open access peer-reviewed chapter
