Organic fertilization strategies and use of Trichoderma in the agronomic performance of green maize

Estratégias de fertilização orgânica e uso do Trichoderma no desempenho agronômico do milho verde

Abstract

There is scientific evidence that Trichoderma positively influences the uptake of soil nutrients by plants, which is essential for the development of more efficient organic fertilization strategies in green maize. Therefore, the objective was to evaluate the efficiency of applying Trichoderma with organic fertilization strategies on the growth and productivity of green . The experiment was conducted at the Piroás Experimental Farm, belonging to the Universidade da Integração Internacional da Lusofonia Afro-Brasileira (UNILAB), located in the municipality of Redenção, Ceará. A randomized complete block design was used with six treatments (T1 - control; T2 - 50% of the cattle manure dose + green manure; T3 - 100% of the cattle manure dose; T4 - Trichoderma + 50% of the cattle manure dose + green manure; T5 - Trichoderma + 25% of the cattle manure dose + green manure; T6 - Trichoderma) and four replications. The combination of Trichoderma with a 50% reduction in the cattle manure dose plus green manure had a positive effect on plant growth, especially in total plant dry mass, when compared to the isolated use of the microorganism. This same combination provided a yield similar to the treatment with the maximum dose of cattle manure. The use of Trichoderma in green maize production was effective in reducing the cattle manure dose by up to 50%, impacting vegetative growth and green maize production when combined with green manure.

Keywords:
Zea Mays L.; fertilization; Mucuna pruriens; fungi

Resumo

Há evidências científicas de que o Trichoderma influencia positivamente a absorção de nutrientes do solo pelas plantas, o que é essencial para o desenvolvimento de estratégias de fertilização orgânica mais eficientes no milho verde. Portanto, o objetivo foi avaliar a eficiência da aplicação de Trichoderma com estratégias de fertilização orgânica no crescimento e produtividade do milho verde. O experimento foi conduzido na Fazenda Experimental de Piroás, pertencente à Universidade da Integração Internacional da Lusofonia Afro-Brasileira (UNILAB), localizada no município de Redenção, Ceará. Foi utilizado um delineamento de blocos casualizados completos com seis tratamentos (T1 - controle; T2 - 50% da dose de esterco bovino + adubo verde; T3 - 100% da dose de esterco bovino; T4 - Trichoderma + 50% da dose de esterco bovino + adubo verde; T5 - Trichoderma + 25% da dose de esterco bovino + adubo verde; T6 - Trichoderma) e quatro repetições. A combinação de Trichoderma com uma redução de 50% na dose de esterco bovino mais a adubação verde teve um efeito positivo no crescimento das plantas, especialmente na massa seca total da planta, quando comparado ao uso isolado do microrganismo. Essa mesma combinação proporcionou uma produtividade semelhante ao tratamento com a dose máxima de esterco bovino. O uso de Trichoderma na produção de milho verde foi eficaz em reduzir a dose de esterco bovino em até 50%, afetando o crescimento vegetativo e a produção de milho verde quando combinado com adubação verde.

Palavras-chave:
Zea Mays L.; fertilização; Mucuna pruriens; fungo

1. Introduction

Maize (Zea mays L.) is considered one of the most important agricultural crops in the world, accounting for about 40% of global cereal production (FAO, 2020). Brazil stands out with 103 million tons of this global production, with an average yield of 5,300 kg ha^-1 (CONAB, 2022). The increase in the supply of cereals in Brazil over the years is related to the increase in productivity, due in part to technological innovations in cultivation, especially in the area of fertilization.

The efficient use of fertilizers in maize production, aimed at increasing nutrient uptake by plants, can improve grain productivity without harming the environment through excessive fertilizer application. This pursuit becomes more challenging in the organic system of family farming, which mostly uses cattle manure as the main source due to its better response compared to other organic materials with higher nutrient content, especially nitrogen (Sampaio et al., 2007). One way to increase the efficiency of cattle manure use is to add residues of nitrogen-fixing plants to the soil to reduce the C:N ratio (Ávila et al., 2020). This increased nitrogen supply from black velvet bean residue, for example, can occur 28 days before the release of nitrogen from cattle manure (Rocha et al., 2019).

Although beneficial, the use of organic fertilization via cattle manure at doses higher than 200 kg ha^-1 N in green maize crop may not fully provide nutrients in the vegetative stages, where their demand is higher in defining crop production components (Rocha et al., 2019). To improve N recovery and prevent its volatilization through organic fertilizers due to the shorter cycle of green maize, it is possible to use efficient microorganisms to enhance plant root growth by releasing substances similar to auxins (Li et al., 2015; França et al., 2017).

The use of microorganisms in agriculture represents a promising alternative to chemical fertilizers due to their environmentally eco-friendly, improving soil structure and biodiversity (Kumar et al., 2024). The genus Trichoderma represents an important group of organisms capable of solubilizing phosphate and making it available to the plant through the release of organic acids, enzymes and phosphatase produced by microorganisms, solubilizing inorganic P and mineralizing organic P (Chagas et al., 2017).

The use of Trichoderma together with organic compounds also has beneficial effects on N absorption and can reduce the recommended dose of organic fertilizer in maize crop without compromising production (Sousa et al., 2018; Ye et al., 2020). Studies present that the use of Trichoderma in combination with organic sources has a positive impact on the development of other crops, such as tomato and sesame (Sani et al., 2020; Khamari et al., 2024).

Therefore, the objective was to evaluate the efficiency of Trichoderma application with organic fertilizer strategies on the growth and productivity of green maize.

2. Material and Methods

2.1. Location and characterisation of the experimental area

The experiment was conducted in two phases, Phase 1 (September 16, 2021 to January 13, 2022) and Phase 2 (January 17 to May 3, 2022), under field conditions at the Piroás Experimental Farm (PEF), belonging to the Universidade da Integração Internacional da Lusofonia Afro-Brasileira (UNILAB), in Redenção, in the state of Ceará, Brazil (4°9’19.39” S and 38°47’41.48” W).

The climate in the region is classified as BSh' (tropical semi-arid climate), characterized by very high temperatures, rainy season in summer and autumn (February to May), intense solar radiation and high evaporation rates (Alvares et al., 2013). The meteorological data during the experimental period were monitored by a data logger (HOBO^® U12-012 Temp/RH/Light/Ext) (Figure 1).

Figure 1
Mean values for maximum (Max) and minimum (Min) temperature and precipitation obtained during the 1^st phase - September 16, 2021, to January 13, 2022 (a) and 2^nd phase - January 17 to May 3, 2022 (b) of the experimental cycle.

The soil texture class of the experimental area was classified as sandy loam. Soil chemical properties were determined according to the methodology of Teixeira et al. (2017) in single samples collected from the soil layer at a depth of 0 to 0.2 meters (m) (Table 1).

Table 1
Chemical and characteristics of the soil at a depth of 0-0.2 m.

Chemical characteristics
pH	OM	P	K+	Na+	Ca+2	Mg+2	Al+3	H+	C/N
-	------g dm-3----		------------------------- (cmolc dm-3) ---------------------						-
6.54	15.62	12	0.13	0.19	6.30	2.20	0.05	1.65	9

• pH H₂O (1:2.5); Extractor Mehlich 1 – P e K; KCl 1 mol L^-1 - Ca^{2 +}. Mg²⁺; Ca (OAc)₂ 0.5 mol L^-1; OM = Organic matter.

2.2. Experimental design and treatments

The experimental design was randomized blocks with six treatments and four replications (T1 - control; T2 - 50% of the cattle manure dose + green manure; T3 - 100% of the cattle manure dose; T4 - Trichoderma + 50% of the cattle manure dose + green manure; T5 - Trichoderma + 25% of the cattle manure dose + green manure; T6 - Trichoderma).

2.3. 1^st phase (area preparation and fertilization)

The species used for green manure fertilization was velvet bean (Mucuna pruriens L.). Sowing took place in the experimental field before the maize crop was planted, only in plots corresponding to the respective treatments. The plots measured 2.2 × 5 m (11 m²), with a spacing of 0.5 × 0.5 m and three seeds per hole. At 120 days after sowing, the velvet bean was cut and incorporated into the soil. Incorporation of the crop took place 35 days before planting the maize crop.

To determine the dry mass, samples were taken from each plot by randomly placing a square of 0.50 × 0.50 m (0.25 m²) where the plants were cut at ground level and placed in paper bags, then placed in an oven at 65 °C. After drying, the material was weighed and the value obtained was converted to t ha^-1, resulting in 5 t ha^-1 of dry mass. It is worth mentioning that in addition to velvet bean, weeds were also collected as they were established in the area, with a dry matter value of 560 kg ha^-1.

Representative samples of the materials used for fertilization (velvet bean, weeds and composted cattle manure) were sent to the laboratory to determine the nutrient contents (Table 2).

Table 2
Chemical characterization of velvet bean, weeds, and cattle manure.

Materials	N	P	K	Ca	Mg	Fe	Cu	Zn
	---------------------------------- g kg-1 --------------------------------------
Velvet bean	21.60	1.74	7.15	-	-	-	-	-
Weeds	20.72	1.06	11.39	-	-	-	-	-
Cattle manure	19.60	4.95	0.67	1.38	3.85	0.55	0.04	0.10

The determination of the dose of organic compounds based on cattle manure was made based on the chemical analysis of the soil (Table 1) and the analysis of organic compounds (Table 2), according to the fertilization recommendations for the State of Ceará (Aquino et al., 1993). Cattle manure fertilization was applied 15 days before maize planting. The doses used were 100% = 20 t ha^-1; 50% = 10 t ha^-1; 25% = 5 t ha^-1. Application was performed according to the treatments, manure was applied on the surface and incorporated into ridges 0.20 m wide and 0.15 m high from the soil surface.

2.4. 2^nd phase (organic fertilization and inoculation of maize)

The maize was planted in February, after fertilization, using the BRS Caatingueiro variety, which has an early cycle (about 100 days). A spacing of 0.75 × 0.33 m was used, with three seeds per hole. Thirteen days after seedling emergence (DAE), thinning was carried out, leaving only one seedling per hole.

During the same period (13 DAE), Trichoderma application was started with four soil applications at 14-day intervals using the commercial product Trichodermil^®, which contains strains of Trichoderma harzianum (STRAIN ESALQ 1306) in its formulation. The dose used was 4.5 L ha^-1, according to the manufacturer's instructions. The product has a concentration of 2 × 10⁹ viable conidia/mL.

2.5. Variables under analysis

At 47 DAE (VT phenological stage - tasseling), the following variables were analyzed: plant height (PH, m), measured with a tape measure from the ground to the end of the male inflorescence; stem diameter (SD, mm), measured 0.05 m above the ground with a digital caliper; number of leaves per plant (NLP); leaf area of the first ear height (LA, cm²), obtained through the expression described and used by Tollenaar (1992), according to Equation 1:

${\text{LA (cm}}^{\text{2}}\text{) = LL × LB × 0}\text{.75}$ (1)

where: LA = Leaf area (cm²); LL = Leaf length (cm); and LB = Leaf blade width (cm).

Still in this period (47 DAE), the plants were then harvested and separated for the determination of root dry mass (RDM) and shoot dry mass (SDM), all obtained after drying in a forced air circulating oven at 65 °C for 72 hours.

When the maize was at phenological stage R3 (milk stage), five ears per plot were harvested and the following variables were determined: average ear mass with straw (AEMWS, g) and average ear mass without straw (AEMWoS, g), determined with an analytical digital balance; ear length (EL, cm), measured with a ruler from end to end of the ear, and ear diameter (ED, mm), measured with a digital caliper at the center of the ear.

For ear yield without straw (EYWoS), the number of plants within 5 m of the plot was counted to estimate the plant density per hectare of marketable ears. The average ear mass without straw was then multiplied by the number of plants with marketable ears to obtain the yield in kg ha^-1.

2.6. Data analysis

The data obtained were subjected to analysis of variance at a p > 0.05 probability level. For mean comparison, the Tukey test was applied at a p > 0.05 probability level. The software used was R 4.1.2 (R Core Team, 2016), using the AgroR 1.3 package Cran R, 2022 (Shimizu et al., 2021).

3. Results

For stem diameter and leaf area variables, no significance was observed among treatments, while for plant height, number of leaves per plant, root dry mass, and shoot dry mass variables, significant influences were observed at a significance level of p ≤ 0.01 and p ≤ 0.05 by the F test (Table 3).

Table 3
Summary of analysis of variance for plant height (PH), stem diameter (SD), number of leaves per plant (NLP), leaf area (LA), root dry mass (RDM) and shoot dry mass (SDM) of green maize as a function of fertilization strategies in soil with Trichoderma.

SV	DF	Mean Square
		PH	SD	NLP	LA	RDM	SDM
Treatments	5	0.10**	4.30ns	1.44**	6249.5ns	11.83*	192.35**
Blocks	3	0.05	6.11	2.04	5078.31	3.56	52.15
Residual	15	0.01	2.69	0.27	4054.05	2.82	30.97
CV (%)	-	6.00	11.10	4.79	13.22	26.96	16.69
Mean	-	1.76	14.77	10.95	481.79	6.23	33.33

• ^*
  significant at F test a 5%;
• ^**
  significant at F test a 1%. SV = source of variation; CV = Coefficient of variation; DF = degrees os freedom; ^nsnot significant.

For plant height (Figure 2a), treatments T2, T3, T4 and T5 did not differ from each other, but were statistically superior to the control (T1) and T6. Treatments T4 and T5 showed a higher number of leaves per plant, statistically superior to the others (Figure 2b).

Figure 2
Mean values for plant height (a) and number of leaves per plant (b) as a function of organic fertilization strategies in soil with and without Trichoderma application. T1 - control; T2 - 50% of the cattle manure dose + green manure; T3 - 100% of the cattle manure dose; T4 - Trichoderma + 50% of the cattle manure dose + green manure; T5 - Trichoderma + 25% of the cattle manure dose + green manure; T6 – Trichoderma. Lowercase letters compare mean values using Tukey test (p ≤ 0.05). Bars represent the standard error of the mean (n = 4).

According to Figure 3a, T3, T4, and T5 were statistically superior to T1, T2, and T6, indicating the beneficial effect of organic and green fertilization associated with Trichoderma, even at the reduced dose. For root dry mass, T2, T3 and T4 gave better statistical results compared to the control and T5 and T6 (Figure 3b).

Figure 3
Mean values for shoot dry mass (a) and root dry mass (b) as a function of organic fertilization strategies in soil with and without Trichoderma application. T1 - control; T2 - 50% of the cattle manure dose + green manure; T3 - 100% of the cattle manure dose; T4 - Trichoderma + 50% of the cattle manure dose + green manure; T5 - Trichoderma + 25% of the cattle manure dose + green manure; T6 – Trichoderma. Lowercase letters compare mean values using Tukey test (p ≤ 0.05). Bars represent the standard error of the mean (n = 4).

For production components, only average ear mass without straw (AEMWoS), and ear yield without straw (EYWoS) showed significance at p ≤ 0.01 and p ≤ 0.05 by F-test (Table 4).

Table 4
Summary of analysis of variance for average ear mass with straw (AEMWS), average ear mass without straw (AEMWoS), ear length (EL), ear diameter (ED), and ear yield without straw (EYWoS) of green maize as a function of fertilization strategies in soil with Trichoderma.

SV	DF	Mean Square
		AEMWS	AEMWoS	EL	ED	EYWoS
Treatments	5	1,977.6ns	1,001.3**	1.09ns	7.36ns	1,967*
Blocks	3	2,465.8	800.49	4.27	2.33	1,573
Residual	15	1,135.4	319.80	2.55	3.29	628,554
CV (%)	-	15.08	11.65	9.42	4.33	11.65
Mean	-	223.4	153.45	16.97	41.88	6,803.29

• ^*
  significant at F test a 5%;
• ^**
  significant at F test a 1%. SV = source of variation; CV = Coefficient of variation; DF = degrees os freedom; ^nsnot significant.

In Figure 4a, the strategy adopted in T4 showed a higher mean value, resulting in an increase of 43.3 g compared to T1, promoting an increase of 34.7% compared to the control, but was statistically similar to T2, T3, T5 and T6. The Figure 4b presents that T4 had the highest mean, exceeding the range of 7,000 kg ha^-1, but it was not statistically different from the other treatments, except for the control (T1), which had an increase of more than 1,900 kg ha^-1, or a superiority of 34% in this respect.

Figure 4
Mean values for ear mass without straw (a) and ear yield without straw (b) as a function of organic fertilization strategies in soil with and without Trichoderma application. T1 - control; T2 - 50% of the cattle manure dose + green manure; T3 - 100% of the cattle manure dose; T4 - Trichoderma + 50% of the cattle manure dose + green manure; T5 - Trichoderma + 25% of the cattle manure dose + green manure; T6 - Trichoderma. Lowercase letters compare mean values using Tukey test (p ≤ 0.05). Bars represent the standard error of the mean (n = 4).

4. Discussion

The lack of stimulation of plant height growth when Trichoderma (T6) was applied alone in maize crop compared to the control treatment (T1) may be related to the ability of this microorganism to compete with maize for nutrients when applied alone. Similarly, Gonilha et al. (2024) found that the isolated use of Trichoderma resulted in the lowest values for maize plant height compared to other treatments that involved different combinations of mineral fertilization and the use of Trichoderma. In contrast, a study conducted by Santos et al. (2021), which evaluated the influence of Trichoderma ssp. on the initial growth of maize, found a growth stimulus in height compared to the control treatment.

However, it is interesting to note that the association of Trichoderma with 25% of the recommended dose of organic fertilizer plus green manure favored height growth. According to Waheed et al. (2020), this improvement in plant growth is also due to the functions of T. harzianum itself, which can promote better root development and phosphorus solubilization associated with increased nutrient availability through organic fertilization. Silva et al. (2023) found a positive effect on maize plant height through the combination of Trichoderma and rock dust, even with reduced doses of NPK. Similarly, Sousa et al. (2023), using cured bovine manure as a fertilizer source in maize crop, also found a positive effect of this organic input on plant height.

This effect may be related to an increased solubilization of microorganisms together with the organic source used. Studies showing the same trend have been reported by Sousa et al. (2023) when fertilizing maize with cured bovine manure in soil with Bacillus aryabhattai, where these authors recorded eight leaves per plant. Similarly, Sousa et al. (2016), who used crab biofertilizer as an organic source in maize crop, also found a higher number of leaves compared to the control treatment. Álvarez et al. (2024) observed a 10% increase in the number of leaves compared to the control when Trichoderma was applied to the maize crop. Santos et al. (2021) reported that volatile compounds produced by Trichoderma improved the maize plants ability to perform Rubisco carboxylation and net photosynthetic rate, resulting in greater biomass accumulation. These results are consistent with those obtained by Mahato et al. (2018), who found a beneficial effect on aboveground dry mass when evaluating the effects of Trichoderma-enriched biofertilizers in wheat crop.

This synergistic effect between Trichoderma and organic fertilizer at a 50% dose of cattle manure plus green manure may be related to the ability of plants to absorb a greater amount of nutrients and promote higher accumulation of dry mass. Vinci et al. (2018) also demonstrated higher dry mass accumulation in maize plants with the activity of Thichoderma plus organic compound. Bader et al. (2020) recorded similar effects of Trichoderma on tomato plants, resulting in an increase in root dry mass. Trichoderma has the ability to produce phytohormones that increase root development by more than 50% in maize crop (Gonilha et al., 2024). This is similar to the results found in this study when comparing T6 to T4 root dry mass.

These results are in agreement with those obtained by Souza et al. (2021), where the same authors observed a positive effect on the ear mass without straw when Trichoderma spp. were applied in maize crop. Similarly, Freire et al. (2022) also observed a higher ear mass without straw in maize crop fertilized with cured bovine manure. Possibly, the amount of rainfall during the study period (Figure 1), combined with the absence of any nutrient input to the plants in the control treatment, resulted in a lower productive performance. On the other hand, all fertilization strategies were efficient, justifying the effect of microorganisms in solubilizing nutrients, promoting a more significant growth and productivity of plants.

The productivity values obtained in this study are lower than those obtained by Favarato et al. (2016), who evaluated the productivity of green maize in a planting system with organic fertilization and recorded a productivity of 10,815 kg ha^-1, and by Arruda et al. (2022), who obtained an average productivity of 12,610 kg ha^-1 for green maize. Sousa et al. (2023), who evaluated the use of Bacillus aryabhattai in maize crop fertilized with cattle manure, also recorded a lower ear productivity than this study (3,098 hg ha^-1).

However, it is noted that the use of Trichoderma combined with organic fertilization at a reduced dose of 50% led an increase in productivity to the control, highlighting the potential of this strategy. Syamsiyah et al. (2023) obtained positive results regarding maize productivity by combining mineral fertilization and Trichoderma, especially when the mineral fertilization dose was reduced.

These results may be related to the ability of Trichoderma to promote improvements in the root system of plants, providing better conditions for water and nutrient absorption and consequently better grain yield (Cruz et al., 2023).

5. Conclusion

The combined strategy of using Trichoderma and green manure resulted in a 50% reduction in the dose of cattle manure, while enhancing the growth parameters and productivity of green maize.

The isolated use of Trichoderma did not improve the agronomic development of maize.

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Publication Dates

• Publication in this collection
  14 Feb 2025
• Date of issue
  2025

History

• Received
  12 July 2024
• Accepted
  29 Nov 2024