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Article

Flowering, Nutritional Status, and Content of Chloroplast Pigments in Leaves of Gladiolus hybridus L. ‘Advances Red’ after Application of Trichoderma spp.

by
Roman Andrzejak
1,* and
Beata Janowska
2,*
1
Department of Phytopathology, Seed Science and Technology, Faculty of Agronomy, Horticulture and Bioengineering, Poznań University of Life Sciences, Dąbrowskiego 159, 60-594 Poznań, Poland
2
Department of Ornamental Plants, Dendrology and Pomology, Faculty of Agronomy, Horticulture and Bioengineering, Poznań University of Life Sciences, Dąbrowskiego 159, 60-594 Poznań, Poland
*
Authors to whom correspondence should be addressed.
Sustainability 2022, 14(8), 4576; https://0-doi-org.brum.beds.ac.uk/10.3390/su14084576
Submission received: 11 March 2022 / Revised: 7 April 2022 / Accepted: 11 April 2022 / Published: 12 April 2022
(This article belongs to the Special Issue Plant-Microbe Interactions and Soil Fertility Status for Enhancing Sustainable Agriculture)
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Abstract

:
In this study, we attempt to assess the influence of Trichoderma spp. on the flowering and nutritional status of Gladiolus hybridus L. ‘Advances Red’, as well as on the content of chlorophyll a + b and carotenoids in the leaves. During both years of the experiment, there was a treatment in which Trichoderma fungi were not used (control), and in another treatment, plants were treated with these fungi. After five weeks of cultivation, when leaf apexes were visible above the surface of the substrate, each plant was irrigated with a suspension (20 mL) of mix of Trichoderma spp. (T. viride Schumach-Tv14, T. harzianum Rifai-Thr2, T. hamatum/Bonord/Bainier-Th15). The treatment of the plants Trichoderma-spp. improved their uptake of macro- (P, K and Ca) and micronutrients (Zn, Fe and B), and increased the chlorophyll a + b and carotenoids in their leaves. Trichoderma spp. accelerated the flowering of Gladiolus hybridus L. ‘Advances Red’ by 10–14 days. The fungi stimulated the elongation of inflorescence shoots and inflorescences, in which the number of flowers increased, but flower diameter did not change. Trichoderma spp. improved the nutrients uptake, chlorophyll a + b and carotenoids, and flowering; hence, Trichoderma spp. treatment is suggested for enhancing inflorescence and inflorescence shoots in Gladiolus hybridus.

1. Introduction

Fungi of the Trichoderma genus are free-living organisms widely distributed in the environment. They occur in all climatic zones and colonize various ecological niches. The most common habitats of Trichoderma spp. are rotting wood and soil, especially the rhizosphere. These fungi produce numerous metabolites which help them interact with plants and other microorganisms [1]. Trichoderma spp. act on bacteria, viruses, and pathogenic fungi through hyperparasitism and antibiosis [2,3]. These fungi are capable of reducing the toxins produced by fungi of the Fusarium genus [4,5]. Recent studies have shown that they may also have complementary properties, which strengthen plants’ protective barriers against insects [6,7]. However, the use of these fungi is somewhat limited by varying levels of their biocontrol activity, which are influenced by environmental conditions. Trichoderma spp. are used to induce systemic resistance in both monocotyledonous and dicotyledonous plants under biotic and abiotic stresses. They are classified as biological control agents (BCA), which are commercially used for the production of plant protection products such as biopesticides and biostimulants. Trichdoderma spp. produce a wide range of bioactive compounds such as enzymes (cellulases, proteases, phosphatases, lipases, xylanases, and amylases) [8], antibiotics, volatile compounds [4,9,10,11], as well as growth regulators [4,12]. Due to their properties, Trichoderma spp. are used as components of microbiological preparations which optimize the composting of raw materials of various origins [8].
Trichoderma spp. have been widely described as plant growth stimulants. However, this trait is isolate-specific rather than species-specific. Individual isolates exhibit various degrees of plant specificity. Growth stimulation is usually manifested by increased root and/or shoot biomass. However, researchers have also described changes in plant morphology and development [12,13]. Growth stimulation may vary considerably due to several limiting factors such as: the type and conditions of cultivation, the dose of the inoculum, and the type of preparation [12,14]. According to Nieto-Jacob et al. [15], communication between plants and Trichoderma spp. comprises the recognition of molecules derived from fungi such as auxins and microbial volatile organic compounds (VOCs). However, this communication is highly dependent on the environment. Contreras-Cornejo et al. [16] suggested that Trichoderma spp. induce growth through an auxin-dependent mechanism. They conducted in vitro biological tests, which proved that Trichoderma virens Gv29.8 and T. atroviride IMI206040 are capable of synthesising indole-3-acetic acid (IAA) and some of its derivatives, due to which the root system develops intensively. According to these authors, many Trichoderma strains are capable of synthesising IAA, but only a few can stimulate plant growth. According to some researchers, Trichoderma spp. stimulate plant growth because they enable plants to absorb more nutrients and support the production of vitamins and growth regulators [17,18,19]. Currently, many Trichoderma bioinoculants are commercially available and blends of strains are becoming more common due to greater consistency of their action [12].
Researchers all over the world have been attempting to determine the influence of Trichoderma spp. on various groups of plants. Most of them have conducted research on functional and edible plants [6,20,21,22]. However, there is not much information about the influence of these fungi on ornamental plants.
Gladiolus hybridus is one of the most popular geophytes grown in the ground and under covers to obtain cut flowers. The quality of cultivars of this species is determined by the colour of flowers, the length and stiffness of inflorescences, as well as the number of flowers developing in the inflorescence [23,24,25].
In this study, it is attempted to assess the influence of Trichoderma spp. on the flowering and nutritional status of Gladiolus hybridus L. ‘Advances Red’, as well as on the content of chlorophyll a + b and carotenoids in the leaves of this plant.

2. Materials and Methods

2.1. Cultivation of Plants

The research was conducted in a greenhouse belonging to the Department of Phytopathology and Seed Science, Poznań University of Life Sciences, Poland. On 29 April 2018 and 2019, Gladiolus hybridus L. ‘Advances Red’ tubers with a circumference of 16–18 cm were planted in openwork boxes with a capacity of 14 L. The boxes were filled with peat substrate (pH 6.2), enriched with the multi-component Osmocote Exact Standard fertilizer (5–6 M) (15:9:12 + 2 MgO + microelement) at rate of 3 g∙dm−3. Five tubers were planted in each container. The plants grown in the greenhouse were watered regularly. After four weeks of cultivation, the plants were treated with the 0.2% multi-component Peters Professional Allrounder fertilizer (20:20:20 + microelements) at an amount of 1 L per container once a week.
There were four treatments in the experiment (research year × Trichoderma spp.). There were fifteen plants in each treatment, five in one replicate. During both years of the experiment there was a treatment in which Trichoderma fungi were not used (control), and in another treatment, plants were treated with these fungi. After five weeks of cultivation, when leaf apexes were visible above the surface of the substrate, each plant was irrigated with a suspension (20 mL) of mix of Trichoderma spp. (T. viride Schumach-Tv14, T. harzianum Rifai-Thr2, T. hamatum/Bonord/Bainier-Th15).

2.2. Inoculum

An inoculum of Trichoderma hamatum, T. harzianum, and T. viride was prepared in sterile plastic Petri dishes with a diameter of 90 mm in a laboratory of the Department of Phytopathology, Seed Science and Technology, Poznań University of Life Sciences. PDA (16 mL) medium was poured into each dish. When it solidified, a 5 mm disc of the nutrient medium overgrown with the mycelium of an appropriate isolate was placed in the central part of the plate. The disc had been cut from the circumference of a 10-day-old culture. Next, the cultures were incubated at 20 °C for three weeks; 20 mL of distilled water was poured onto the sporulating cultures, and the resulting suspension was poured into a flask. A spore suspension of the three tested Trichoderma isolates was prepared from a three-week culture. Trichoderma isolates were flooded with 20 mL of sterilized distilled water and scraped with a sterile glass rod. The suspension was filtered, and the spore concentration of the three Trichoderma species in the mixture was adjusted to a concentration of 106 per mL using a hemocytometer under light microscopy.

2.3. Parameters

When two lower flowers developed in the spike, the shoots of the inflorescences were cut above the second leaf. The following parameters were measured: earliness of flowering (measured with the number of days from the planting of tubers to the harvesting of inflorescences), the length of the inflorescence shoot measured from the ground surface, the length of the inflorescence, the inflorescence diameter, and the number of flowers and buds. The content of macronutrients (nitrogen—N, phosphorus—P, potassium—K, calcium—Ca, magnesium—Mg), micronutrients (iron—Fe, manganese—Mn, zinc—Zn, copper—Cu, boron—B), chlorophyll a + b and carotenoids in the leaves was also measured. At the end of the experiment (after 17 weeks of cultivation), the percentage of root colonization by Trichoderma spp. was determined.

2.4. Content of Chloroplast Pigments

In order to evaluate the biochemical changes in the leaves of Gladiolus hybridus L. ‘Advances Red’, the content of chlorophyll a + b and carotenoids was determined. Leaf tips with a length of 5 cm were collected for analyses. In order to measure the content of chloroplast pigments, 200-mg samples were collected from the leaves of the plants cultivated with or without Trichoderma spp.
The levels of pigments were measured with the methods described by Hiscox and Israelstam [26]. The pigments were extracted with dimethyl sulphoxide (DMSO) without tissue maceration. Weighed portions (100 mg) were treated with 5 cm3 DMSO and incubated in a water bath at 65 °C for 60 min. The levels of pigments in the extract were measured spectrophotometrically at adequate wavelengths. The absorbance of the extract was measured at a wavelength of 663 nm for chlorophyll a, at 645 nm for chlorophyll b, and at 470 nm for carotenoids. The content of the pigments was calculated by means of Arnon’s [27] formulae and expressed as mg/g fresh weight.

2.5. Content of Macro- and Microelements

Leaf tips with a length of 10 cm were collected for chemical analyses in each treatment. The leaves were dried at a temperature of 45–50 °C and then ground. In order to measure the total content of N, P, K, Ca, and Mg, the leaves were mineralised in concentrated sulphuric acid (H2SO4). The following methods were used to measure the content of the nutrients: total N—Kjeldahl digestion with distillation in a Parnas-Wagner apparatus; P—the colorimetric method with ammonium molybdate (after Schillak); K, Ca, and Mg—atomic absorption spectrometry (AAS).
In order to measure the total Fe, Mn, Zn, and Cu content, the leaves were mineralised in a mixture of nitric (HNO3) and perchloric acids (HClO4) (3:1, v/v). In order to measure the Na content, they were mineralised in concentrated sulphuric acid (H2SO4) [28]. After the mineralisation, the Na, Fe, Mn, Zn, and Cu content was measured with the AAS method (in a Carl Zeiss Jena apparatus).

2.6. Root Colonization

At the end of the experiment, after 17 weeks of cultivation, the shoots were removed and the tubers were dug up with the roots. Then, they were rinsed in water until the substrate was removed. Root samples were collected and cut into 1-cm-long pieces. The samples were surface-disinfected by immersion in a 2% sodium hypochlorite (NaOCl) solution for 2 min. Root fragments (5 in each sample) were placed in a Petri dish with PDA medium and incubated at 20 °C for 14 days. Then, they were placed on sterile filter paper and dried in a laminar airflow cabinet. The percentage of root colonization was assessed on the basis of the number of roots colonized by Trichoderma spp.

2.7. Data Analysis

The results were analysed statistically with two-way analysis of variance. The experiment was set up in a randomised complete block design. The averages were grouped by means of the Duncan test at a significance level of α = 0.05. The Statistica 13.3 program (StatSoft Polska, Kraków, Poland) was used for the calculations.

3. Results and Discussion

The experiment showed that in the treatments with Trichoderma spp., 46.6% and 48.2% of the roots of the Gladiolus hybridus ‘Advances Red’ plants were colonized by these fungi (Figure 1).
Prisa et al. [29] indicated that Trichoderma spp. are free-living fungi, which are commonly found in soil and root ecosystems. Some strains colonize plant roots heavily and for a long time by penetrating the first layers of the epidermis. Research conducted so far has shown differences in the degree of root colonization by fungi of the Trichoderma genus. Janowska et al. [30] conducted a study on Freesia reflacta ‘Argentea’ and observed a lower percentage of root colonization by Trichoderma spp., i.e., 32% and 33%. Likewise, Andrzejak et al. [31] observed that the percentage of colonization of the roots of Begonia × tuberhybrida ‘Picotee Sunburst’ by Trichoderma spp. amounted to 30.5%, 29.5%, and 30.0%. On the other hand, Prisa et al. [29] conducted a study on Limonium sinuatum and found that the colonization of plant roots by fungi of the Trichoderma genus may be very high, i.e., 100%. According to Błaszczyk et al. [32], Trichoderma spp. colonize not only the outer layers of the roots of herbaceous plants and trees in the rhizosphere, but they also have the ability to penetrate the roots and colonize them inside, or they may occur as endophytes. The researchers conducted their study on Triticum aestivum. The preliminary analysis of morphological, anatomical, physiological, and metabolomic changes showed that the plants did not exhibit a clear reaction to Trichoderma fungi. This might mean that the changes occurring in plants depend on both the Trichoderma species/strain and variety. According to Souz et al. [33], plant–microbiota interactions occurring in the rhizosphere are key determinants of plant health, productivity, and soil fertility. Plant roots synthesise metabolites, which are recognised by microorganisms, which respond with signals initiating microbial colonization [34]. Plant roots also secrete sucrose as a source of energy to support microbial colonization [35,36].
The study showed that the earliness of flowering of the plants depended only on the use of Trichoderma spp. during the cultivation (Figure 2 and Figure 3). Regardless of the year of the study, the plants treated with fungi of the Trichoderma genus started flowering 10 days earlier on average. The comparison of the interactions showed that in both years of the study, the plants grown with Trichoderma spp. flowered 10 and 14 days earlier than the control plants grown without the fungi. The earliness of flowering of ornamental plants is a very important parameter, due to which it is possible to schedule the harvest. For this reason, it is essential to know the reactions of individual plant species and their cultivars to treatments. So far, research has shown that many species begin to flower earlier after the treatment with Trichoderma spp. Janowska et al. [30] observed that when Trichoderma spp. was applied to Freesia refracta ‘Argentea’ grown for winter flowers without assimilation lighting, the plants started flowering about one week earlier. Andrzejak et al. [31] found that Trichoderma spp. slightly accelerated the flowering of Begonia × tuberhybrida ‘Picotee Sunburst’ when the plants were fed with the fertilizer at a concentration of 0.2%. A higher concentration of the fertilizer accelerated the flowering of the plants by 8.7 days.
Trichoderma spp. stimulated the growth of inflorescence shoots from the ‘Advances Red’ cultivar. The shoots were significantly longer (by 9.8%), and had 10% longer inflorescences, in which the number of flowers increased by 12.6% (Figure 3 and Figure 4A,C). However, the flowers in all treatments had similar diameters (Figure 4D). These results are partly similar to the findings of the study conducted by Sisodia et al. [37], who applied Trichoderma spp. to eight Gladiolus sp. cultivars. As a result, the length of the spike and the duration of flowering increased, but the number of flowers per spike did not change. On the other hand, da Cruz et al. [38] did not observe any effect of Trichoderma spp. on the quality of Gladiolus ‘Peter’s Pear’ inflorescences—there were no changes in the length of the inflorescence shoot, the length of the inflorescence, or the number of flowers. The study by Andrzejak et al. [31] showed that Trichoderma spp. stimulated the development of buds and flowers of Begonia × tuberhybrida ‘Picotee Sunburst’ and affected their size. The plants bloomed most intensively and had the biggest flowers when they were fed with the fertilizer at a concentration of 0.3%. The results of our study are also in line with the findings of the research conducted on Freesia refracta ‘Argentea’ by Janowska et al. [30], who observed that Trichoderma spp. stimulated the development of lateral inflorescence shoots and flowers, especially in the plants which were exposed to assimilation lighting during cultivation. According to Prisa [39], fungi of the Trichoderma genus also stimulate the flowering of Pachyphytum oviferum and Crassula falcata.
The comparison of the content of chlorophyll a + b and carotenoids in our study showed that in the treatments ‘Trichoderma spp. Yes’, the concentrations of both pigments in the leaves increased significantly (chlorophyll a + b—by 66.7%, and carotenoids—by 33.3%) (Figure 3 and Figure 5A,B). The results of our study indicate that the photosynthetic capacity of the ‘Advances Red’ cultivar improved. Andrzejak et al. [30] observed that Trichoderma spp. also stimulated the production of chlorophyll in Begonia × tuberhybrida ‘Picotee Sunburst’. The content of this pigment was reflected by the leaf greenness index (SPAD). According to Harman et al. [40], the authors of numerous studies documented the fact that various endophytic Trichoderma spp. improved the photosynthetic capacity of various plant species by increasing the content of photosynthetic pigments or the expression of genes regulating the biosynthesis of chlorophyll, proteins of the light-harvesting complex, or components of the Calvin cycle. The colonization of the roots of crops by the Trichoderma spp. fungi increases the regulation of genes and pigments, which improve photosynthesis. When plants are exposed to physiological or environmental stress, they lose their photosynthetic ability, because their photosystems are damaged and various cellular processes are disordered by reactive oxygen species (ROS). ROS generation is central to all living organisms, which is due to normal cellular metabolism and also in response to environmental stresses. However, a perturbed ROS homoeostasis causes oxidative damage to macromolecules (proteins, nucleic acids and lipids), which further leads to irreversible changes in protein structure and function. A good balance between ROS generation and the antioxidant defense system protects photosynthetic machinery, maintains membrane integrity, and prevents damage to nucleic acids and proteins. Notably, the antioxidant defense system not only scavenges ROS, but also regulates the ROS titer for signaling [41,42]. However, some Trichoderma strains activate the biochemical pathways which reduce ROS to less harmful molecules. This mechanism, as well as other mechanisms, make plants more resistant to biotic and abiotic stresses. Apart from that, when photosynthetic rates increase, more carbon dioxide (CO2) is taken up from the atmosphere. Most studies on this issue were conducted on edible crops [20,21,22]. However, as demonstrated by the results from research, Trichoderma spp. also stimulate the formation of photosynthetic pigments (chlorophyll, carotenoids) in ornamental plants. Carotenoids have very important functions in the plant world. They are responsible for the stability of lipid membranes, they take part in the accumulation of light during photosynthesis, and they provide protection against photooxidation, which is caused by reactive oxygen species formed during the excitation of chlorophyll during photosynthesis [43,44]. The antioxidative effect of carotenoids on lipid membranes depends on their orientation, location, and organisation in the membranes. Polar and non-polar carotenoids have different effects on the structure and physiology of tissues. For example, astaxanthin, a polar substance, reduces lipid peroxidation by maintaining rigidity of the membrane structure [45]. Carotenoids are highly active against reactive oxygen species and free radicals [43]. Metwally and Al-Amri [46] observed that the combined inoculation of Allium cepa with mycorrhizal fungi (AM) and biocontrol fungus (Trichoderma viride) not only improved the growth parameters of onion (fresh and dry weight, length of roots and shoots, leaf area), but also increased the content of chlorophyll, carotenoids, and total pigments in the leaves. Abdel-Fattah et al. [47] found that the spraying of Oryza plants with a suspension of T. harzianum spores significantly increased the content of photosynthetic pigments (chlorophyll a + b and carotenoids) in the leaves. One of the Trichoderma strains (T. azevedoi CEN1241) also increased the content of chlorophyll and carotenoids in the leaves of Lectuca sativa [48]. Hosseinzeynali et al. [49] observed that the treatment of Pistacia with T. harzianum, T. viride, and a mixture of these fungi increased the content of total chlorophyll to 73.2%, 171%, and 59.2%, and the content of carotenoids to 77.6, 3.80, and 64.8%, respectively.
The comparison of the content of macronutrients in the leaves of the ‘Advances Red’ cultivar after the application of Trichoderma spp. showed that these fungi significantly influenced the uptake of P, K, and Ca by the plants (Table 1, Figure 3). In both years of the research, the leaves which grew from the plants treated with Trichoderma spp. had significantly higher content of P, K, and Ca than the leaves of the control plants. There were similar results of the study conducted by Janowska et al. [30] on Freesia reflacta ‘Argentea’. The authors observed that Trichoderma spp. stimulated the uptake of P and Ca in the underexposed plants, whereas in the plants exposed to assimilation lighting, the fungi additionally stimulated the uptake of K. The application of microorganisms improves the uptake of nutrients. Due to this effect, the amount of organic fertilizers can be reduced, which is in line with the latest trends in horticultural and agricultural practices. Biofertilizers based on microorganisms are an alternative—it is possible to maintain high production while reducing the influence on the natural environment [46,49,50,51]. Biofertilizers can be used as a supplement or alternative to chemical fertilizers in sustainable plant production [45]. Metwally [50] observed that both mycorrhizal (AM) and T. viride fungi were compatible with each other. The researcher applied them in combination to Allium cepa and found that they not only improved the biochemical parameters of the plants, such as the content of soluble carbohydrates, proteins, free amino acids, acid and alkaline phosphatases, but also increased the content of minerals and nutrients (N, P, K+, Ca2+, Mg2+, and Zn). When the plants were inoculated with AM, T. viride and the combination of both fungal biostimulants, the P content in the shoots increased by 67%, 49%, and 112%, respectively. Hosseinzeynali et al. [49] observed that the application of T. harzianum, T. viride, and a mixture of these fungi increased the K content in Pistacia leaves by 20%, whereas T. harzianum increased the P content by 14% and the Ca content by 40%.
The comparison of the content of microelements in the leaves of ‘Advances Red’ cultivar after the application of Trichoderma spp. showed that these fungi significantly influenced the uptake of Zn, Fe, and B by the plants (Table 2, Figure 3). In both years of the research, the leaves of the plants treated with Trichoderma spp. had significantly higher Zn, Fe, and B levels than the leaves of the control plants. There were no differences between the years of the research. Micronutrients play a key role in the metabolic and physiological processes of plants. They affect the yield quality to a greater extent [52] than the yield volume. Micronutrients are components of proteins and have catalytic functions [53]. The results of our study were in line with the findings of the research on Begonia × tuberhybrida ‘Picotee Sunburst’ conducted by Andrzejak et al. [31], who observed that Trichoderma spp. also stimulated the uptake of micronutrients (Zn, Fe, and B) in this ornamental plant. The higher the concentration of the fertilizer was, the higher the content of these micronutrients was. Janowska et al. [30] found that Trichoderma spp. stimulated the uptake of Fe, Mn, and Zn in both illuminated and non-illuminated Freesia reflacta ‘Argentea’ plants. The researchers also observed that the combination of illumination and Trichoderma spp. stimulated the Cu uptake. Trichoderma spp. also stimulate the uptake of micronutrients in vegetables. For example, the content of Cu, Fe, Mn, and Zn in the roots, shoots, and fruits of Solanum lycopersicum [54] and Cucumis sativus [17] increased significantly in response to T. harzianum inoculation. However, da Santiago et al. [55] observed that Trichoderma asperellum competed for Cu, Mn, and Zn and decreased the concentration of these elements in plants. Borrero et al. [56] observed that the leaves of Solanum lycopersicum grown in the presence of T. harzianum T34 had a lower Fe content. Triticum aestivum grown on a calcareous substrate and inoculated with T. asperellum had lower concentrations of Cu, Mn, and Zn [55]. The authors suggested that the lower concentrations of these elements in the plants were caused by the competition between the plants and Trichoderma spp. There were interesting results of the research conducted on Solanum lycopersicum ‘Gružanski zlatni’ by Vukelić et al. [57]. When the researchers applied Trichoderma spp. to this cultivar, the nitrogen balance index in the leaves decreased as a result of the increase in the content of flavonols in the epidermis and the decrease in the content of chlorophyll. According to the researchers, this effect was caused by switching from primary to secondary metabolism. The quality of the fruit changed because the content of flavonoids had increased, the content of starch had decreased, and the bioaccumulation index (BI) for iron (Fe) and chromium (Cr) had increased, whereas the BI for heavy metals—nickel (Ni) and lead (Pb)—had decreased. According to the researchers, the higher expression of the swolenin gene in the roots of the more sensitive cultivar of Solanum lycopersicum indicated better colonization, which correlated with the positive effects of Trichoderma spp. Although many studies proved that Trichoderma spp. significantly influence the uptake of nutrients, further research is necessary to investigate thoroughly the effects of the inoculation of individual plant species and cultivars with these fungi.

4. Conclusions

The treatment of the plants Trichoderma spp. improved their uptake of macro- (P, K and Ca) and micronutrients (Zn, Fe and B), and increased the chlorophyll a + b (by 66.7%) and carotenoids (33.3%) in their leaves. Trichoderma spp. accelerated the flowering of Gladiolus hybridus L. ‘Advances Red’ by 10–14 days. The fungi stimulated the elongation of inflorescence shoots (by 9.8%) and inflorescences (by 9.8%), in which the number of flowers increased (by 12.6%), but the flowers had similar diameters. Trichoderma spp. improved the nutrients uptake, chlorophyll a + b and carotenoids, and flowering; hence, Trichoderma spp. treatment is suggested for enhancing inflorescence and inflorescence shoots in Gladiolus hybridus.

Author Contributions

Conceptualization, R.A.; methodology, R.A.; formal analysis, R.A. and B.J.; funding acquisition, R.A. and B.J.; writing—original draft, R.A. and B.J.; writing—review and editing, R.A. and B.J. All authors have read and agreed to the published version of the manuscript.

Funding

The publication was co-financed within the framework of the Ministry of Science and Higher Education program as “Regional Initiative Excellence” in 2019–2022, Project No. 005/RID/2018/19, financing amount: 12,000,000 PLN.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Chandra, H.; Kumari, P.; Prasad, R.; Chandra Gupta, S.; Yadav, S. Antioxidant and antimicrobial activity displayed by a fungal endophyte Alternaria alternata isolated from Picrorhiza kurroa from Garhwal Himalayas, India. Biocat. Agric. Biotechnol. 2021, 33, 101955. [Google Scholar] [CrossRef]
  2. Hewedy, O.A.; Abdel Lateif, K.S.; Seleiman, M.F.; Shami, A.; Albarakaty, F.M.; MEl-Meihy, R. Phylogenetic diversity of Trichoderma strains and their antagonistic potential against soil-borne pathogens under stress conditions. Biology 2020, 9, 189. [Google Scholar] [CrossRef] [PubMed]
  3. Wojtkowiak-Gebarowska, E. Mechanizmy zwalczania fitopatogenow glebowych przez grzyby z rodzaju Trichoderma. Post. Mikrobiol. 2006, 45, 261–273. [Google Scholar]
  4. Benìtez, T.; Rincòn, A.M.; Limòn, M.C.; Codòn, A.C. Biocontrol mechanisms of Trichoderma strains. Int. Microbiol. 2004, 7, 249–260. [Google Scholar]
  5. Świerczyńska, I.; Korbas, M.; Horoszkiewicz-Janka, J.; Danielewicz, J. Antagonistyczne oddziaływanie Trichoderma viride na patogeny z rodzaju Fusarium w obecności biopreparatów (Antagonistic effect of Trichoderma viride on pathogenic fungi of the genus Fusarium in the presence of biopreparations). J. Res. Appl. Agric. Eng. 2011, 56, 157–160. [Google Scholar]
  6. di Lelio, I.; Coppola, M.; Comite, E.; Molisso, D.; Lorito, M.; Woo, S.L.; Pennacchio, F.; Rao, R.; Digilio, M.C. Temperature differentially influences the capacity of Trichoderma species to induce plant defense responses in Tomato against insect pests. Front. Plant Sci. 2021, 12, 678830. [Google Scholar] [CrossRef]
  7. Poveda, J. Trichoderma as biocontrol agent against pests: New uses for a mycoparasite. Biol. Control 2021, 159, 104634. [Google Scholar] [CrossRef]
  8. Kosicka, D.; Wolna-Maruwka, A.; Trzeciak, M. Aspekty stosowania Trichoderma sp. w ochronie roślin i rozkładzie materii organicznej (Aspects of the use of Trichoderma sp. in crop protection and distribution of organic matter). Kosmos 2014, 63, 635–642. [Google Scholar]
  9. Vey, A.; Hoagland, R.E.; But, T.M. Toxic metabolites of fungal biocontrol agents. In Fungi as Biocontrol Agents: Progress, Problems and Potential; Butt, T.M., Jackson, C., Magan, N., Eds.; CABI Publishing: Egham, UK, 2001; pp. 311–346. [Google Scholar]
  10. Howell, C.R. Understanding the mechanisms employed by Trichoderma virens to effect biological control of cotton diseases. Phytopathology 2006, 96, 178–180. [Google Scholar] [CrossRef] [Green Version]
  11. Reino, J.L.; Guerrero, R.F.; Hernández-Galán, R.; Collado, I.G. Secondary metabolites from species of the biocontrol agent Trichoderma. Phytochem. Rev. 2008, 7, 89–123. [Google Scholar] [CrossRef]
  12. Stewart, A.; Hill, R. Applications of Trichoderma in plant growth promotion. In Biotechnology and Biology of Trichoderma; Gupta, V.K., Schmoll, M., Herrera-Estrella, A., Upadhyay, R.S., Druzhinina, I., Tuohy, M.G., Eds.; Elsevier: Amsterdam, The Netherlands, 2014; pp. 415–428. [Google Scholar] [CrossRef]
  13. Zin, N.A.; Badaluddin, N.A. Biological functions of Trichoderma spp. for agriculture applications. Ann. Agric. Sci. 2020, 65, 168–178. [Google Scholar] [CrossRef]
  14. Hoyos-Carvajal, L.; Orduz, S.; Bissett, J. Growth stimulation in bean (Phaseolus vulgaris L.) by Trichoderma. Biol. Control 2009, 51, 409–416. [Google Scholar] [CrossRef]
  15. Nieto-Jacobo, M.F.; Steyaert, J.M.; Salazar-Badillo, F.B.; Nguyen, D.V.; Rostás, M.; Braithwaite, M.; de Souza, J.T.; Jimenez-Bremont, J.F.; Ohkura, M.; Stewart AMendoza-Mendoza, A. Environmental growth conditions of Trichoderma spp. affects indole acetic acid derivatives, volatile organic compounds, and plant growth promotion. Front. Plant Sci. 2017, 8, 102. [Google Scholar] [CrossRef] [Green Version]
  16. Contreras-Cornejo, H.A.; Macías-Rodríguez, L.; Cortés-Penagos, C.; López-Bucio, J. Trichoderma virens, a plant beneficial fungus, enhances biomass production and promotes lateral root growth through an auxin-dependent mechanism in Arabidopsis. Plant Physiol. 2009, 149, 1579–1592. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  17. Yedidia, I.; Srivastava, A.K.; Kupalnik, Y.; Chet, I. Effect of Trichoderma harzianum on microelement concentrations and increased growth of Cucumber plants. Plant Soil. 2001, 235, 235–242. [Google Scholar] [CrossRef]
  18. Harman, G.E.; Howell, C.R.; Viterbo, A.; Chet, I.; Lorito, M. Trichoderma species–opportunistic, avirulent plant symbionts. Nat. Rev. Microbiol. 2004, 2, 43–56. [Google Scholar] [CrossRef]
  19. Lorito, M.; Woo, S.; Harman, G.; Monte, E. Translational research on Trichoderma: From ‘omics to the field. Annu. Rev. Phytopathol. 2010, 48, 395–417. [Google Scholar] [CrossRef] [Green Version]
  20. Simkin, A.J.; Gaffe, J.; Alcaraz, J.P.; Carde, J.P.; Bramley, P.M.; Fraser, P.D.; Kuntz, M. Fibrillin influence on plastid ultrastructure and pigment content in tomato fruit. Phytochemistry 2007, 68, 1545–1556. [Google Scholar] [CrossRef]
  21. Langi, P.; Kiokias, S.; Varzakas, T.; Proestos, C. Carotenoids: From plants to food and feed industries. Methods Mol. Biol. 2018, 1852, 57–71. [Google Scholar]
  22. Meléndez-Martínez, A.J.; Mandić, A.I.; Bantis, F.; Böhm, V.; Borge, G.I.A.; Brnĉiĉ, M.; Bysted, A.; Cano, M.P.; Dias, M.G.; Elgersma, A.; et al. A comprehensive review on carotenoids in foods and feeds: Status quo, applications, patents, and research needs. Crit. Rev. Food Sci. Nutr. 2022, 62, 1999–2049. [Google Scholar] [CrossRef]
  23. Havale, V.B.; Tawar, R.V.; Hage, N.D.; Kakad, G.J.; Fathepurkar, S.C.; Sable, A.S. Effect of growth regulators and chemicals on growth and flowering of Gladiolus. Asian J. Hortic. 2008, 3, 93–94. [Google Scholar]
  24. Janowska, B.; Andrzejak, R.; Kosiada, T.; Kwiatkowska, M.; Smolińska, D. The flowering and nutritional status of Gladiolus hybridus cv. Black Velvet following a cytokinin treatment. J. Elementol. 2018, 23, 1119–1128. [Google Scholar] [CrossRef]
  25. Janowska, B.; Andrzejak, R.; Kosiada, T.; Kwiatkowska, M.; Smolińska, D. The flowering and nutritional status of Gladiolus hybridus ‘Black Velvet’ following gibberellin treatment. Hort. Sci. 2018, 45, 205–210. [Google Scholar] [CrossRef] [Green Version]
  26. Hiscox, J.D.; Israelstam, G.F. A method for the extraction of chlorophyll from leaf tissue without maceration. Can. J. Bot. 1979, 57, 1332–1334. [Google Scholar] [CrossRef]
  27. Arnon, D. Copper enzymes in isolated chloroplasts. polyphenoloxidase in Beta vulgaris. Plant Physiol. 1949, 24, 1–15. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  28. Kamińska, W.; Kardasz, T.; Strahl, A.; Bałucka, T.; Walczak, K.; Filipek, P. The methods of analysis in chemical-agricultural station. Part II. In Analisis of Plants; IUNG: Puławy, Poland, 1972. [Google Scholar]
  29. Prisa, D.; Sarrocco, S.; Forti, M.; Burchi, G.; Vannacci, G. Endophytic ability of Trichoderma spp. as inoculants for ornamental plants innovative substrates. J. Plant Pathol. 2013, 86, 169–174. [Google Scholar]
  30. Janowska, B.; Andrzejak, R.; Kosiada, T. The influence of fungi of the Trichoderma genus on the flowering of Freesia refracta Klatt ‘Argentea’ in winter. Hort. Sci. 2020, 47, 203–210. [Google Scholar] [CrossRef]
  31. Andrzejak, R.; Janowska, B.; Reńska, B.; Kosiada, T. Effect of Trichoderma spp. and fertilization on the flowering of Begonia × tuberhybrida Voss. ‘Picotee Sunburst’. Agronomy 2021, 11, 1278. [Google Scholar] [CrossRef]
  32. Błaszczyk, L.; Witaszak, N.; Basińska-Barczak, A.; Marczak, Ł.; Sawikowska, A.; Perlikowski, D.; Kosmala, A. Reakcja roślin pszenicy zwyczajnej (Triticum aestivum L.) na kolonizację korzeni przez grzyby Trichoderma. Biul. IHAR 2019, 285, 129–230. [Google Scholar]
  33. Souza, R.; Ambrosini, A.; Passaglia, L.M. Plant growth-promoting bacteria as inoculants in agricultural soils. Genet. Mol. Biol. 2015, 38, 401–419. [Google Scholar] [CrossRef]
  34. Berg, G. Plant-microbe interactions promoting plant growth and health: Perspectives for controlled use of microorganisms in agriculture. Appl. Microbiol. Biotechnol. 2009, 84, 11–18. [Google Scholar] [CrossRef] [PubMed]
  35. Druzhinina, I.S.; Seidl-Seiboth, V.; Herrera-Estrella, A.; Horwitz, B.A.; Kenerley, C.M.; Monte, E.; Mukherjee, P.K.; Zeilinger, S.; Grigoriev, I.V.; Kubicek, C.P. Trichoderma: The genomics of opportunistic success. Nat. Rev. Microbiol. 2011, 9, 749–759. [Google Scholar] [CrossRef]
  36. Vargas, W.A.; Crutcher, F.K.; Kenerley, C.M. Functional characterisation of a plant-like sucrose transporter from the beneficial fungus Trichoderma virens. Regulation of the symbiotic association with plants by sucrose metabolism inside the fungal cells. New Phytol. 2011, 189, 777–789. [Google Scholar] [CrossRef] [PubMed]
  37. Sisodia, A.; Pal, A.; Singh, A.K. Varietal response and effect of Trichoderma on flowering in gladiolus. J. Pharmacogn. Phytochem. 2018, 7, 3658–3660. [Google Scholar]
  38. da Cruz, L.; Ludwig, F.; Steffen, G.; Joseila Maldaner, J. Development and quality of gladiolus stems with the use of vermicompost and Trichoderma sp. in substrate. Ornam. Hortic. 2018, 24, 70–77. [Google Scholar] [CrossRef] [Green Version]
  39. Prisa, D. Trichoderma harzianum: Biocontrol to Rhizoctonia solani and biostimulation in Pachyphytum oviferum and Crassula falcata. WJARR 2019, 3, 11–18. [Google Scholar]
  40. Harman, G.E.; Doni, F.; Khadka, R.B.; Uphoff, N. Endophytic strains of Trichoderma increase plants’ photosynthetic capability. J. Appl. Microbiol. 2021, 130, 529–546. [Google Scholar] [CrossRef]
  41. Yadav, S.; Kushwaha, H.R.; Kumar, K.; Verma, P.K. Comparative structural modeling of a monothiol GRX from chickpea: Insight in iron-sulfur cluster assembly. Int. J. Biol. Macromol. 2012, 51, 266–273. [Google Scholar] [CrossRef]
  42. Hasanuzzaman, M.; Bhuyan, M.H.; Parvin, K.; Bhuiyan, T.F.; Anee, T.I.; Nahar, K.; Hossen, M.; Zulfiqar, F.; Alam, M.; Fujita, M. Regulation of ROS metabolism in plants under environmental stress: A review of Recent Experimental Evidence. Int. J. Mol. Sci. 2020, 21, 8695. [Google Scholar] [CrossRef]
  43. Andersson, S.C. Carotenoids, Tocochromanols and Chlorophylls in Sea Buckthorn Berries (Hippophae rhamnoides) and Rose Hips (Rosa sp.). Ph.D. Thesis, Swedish University of Agricultural Sciences, Alnarp, Sweden, 2009; pp. 15–20. [Google Scholar]
  44. Simkin, A.J. Carotenoids and apocarotenoids in planta: Their role in plant development, contribution to the flavour and aroma of fruits and flowers, and their nutraceutical benefits. Plants 2021, 10, 2321. [Google Scholar] [CrossRef]
  45. Widomska, J.; Kostecka-Gugała, A.; Latowski, D.; Gruszecki, W.I.; Strzałka, K. Calorimetric studies of the effect of cis-carotenoids on the thermotropic phase behavior of phosphatidylcholine bilayers. Biophys. Chem. 2008, 140, 108–114. [Google Scholar] [CrossRef] [PubMed]
  46. Metwally, R.A.; Al-Amri, S. Individual and interactive role of Trichoderma viride and arbuscular mycorrhizal fungi on growth and pigment content of onion plants. Lett. Appl. Microbiol. 2020, 70, 79–86. [Google Scholar] [CrossRef] [PubMed]
  47. Abdel-Fattah, G.M.; Shabana, Y.M.; Adel EIsmail, A.E.; Rashad, Y.M. Trichoderma harzianum: A biocontrol agent against Bipolaris oryzae. Mycopathologia 2007, 164, 81–89. [Google Scholar] [CrossRef] [PubMed]
  48. da Silva, L.R.; Valadares-Inglis, M.C.; Peixoto, G.H.; de Luccas, B.E.; Muniz, P.H.; Magalhães, D.M.; Moraes, M.C.; de Mello, S.C. Volatile organic compounds emitted by Trichoderma azevedoi promote the growth of lettuce plants and delay the symptoms of white mold. Biol. Control 2021, 152, 104447. [Google Scholar] [CrossRef]
  49. Hosseinzeynali, A.; Abbaszadeh Dahaji, P.; Alaei, H.; Hosseinifard, J.; Akhgar, A. Effect of Trichoderma on growth and nutrition of Pistachio trees under ommon garden conditio. J. Soil Biol. 2021, 8, 115–129. [Google Scholar]
  50. Metwally, R.A. Arbuscular mycorrhizal fungi and Trichoderma viride cooperative effect on biochemical, mineral content, and protein pattern of onion plants. J. Basic Microbiol. 2020, 60, 712–721. [Google Scholar] [CrossRef]
  51. Chandra, H.; Kumari, P.; Bisht, R.; Prasad, R.; Yadav, S. Plant growth promoting Pseudomonas aeruginosa from Valeriana wallichii displays antagonistic potential against three phytopathogenic fungi. Mol. Biol. Rep. 2020, 47, 6015–6026. [Google Scholar] [CrossRef]
  52. Khosa, S.S.; Younis, A.; Rayit, A.; Yasmeen, S.; Riaz, A. Effect of foliar application of macro and micro nutrients on growth and flowering of Gerbera jamesonii L. AEJAES 2011, 11, 736–757. [Google Scholar]
  53. Lahijie, M.F. Application of micronutrients FeSO4 and ZnSO4 on the growth and development of Gladiolus variety ‘Oscar’. IJACS 2012, 4, 718–720. [Google Scholar]
  54. Khan, M.Y.; Haque, M.M.; Molla, A.H.; Rahman, M.M.; Alam, M.Z. Antioxidant compounds and minerals in tomatoes by Trichoderma-enriched biofertilizer and their relationship with the soil environments. JIA 2017, 16, 691–703. [Google Scholar] [CrossRef]
  55. de Santiago, A.; Quintero, J.M.; Avilés, M.; Delgado, A. Effect of Trichoderma asperellum strain T34 on iron, copper, manganese, and zinc uptake by wheat grown on a calcareous medium. Plant Soil. 2011, 342, 97–104. [Google Scholar] [CrossRef]
  56. Borrero, C.; Trillas, M.I.; Delgado, A.; Avilés, M. Effect of ammonium/nitrate ratio in nutrient solution on control of Fusarium wilt of tomato by Trichoderma asperellum T34. Plant Pathol. 2012, 61, 132–139. [Google Scholar] [CrossRef]
  57. Vukelić, I.D.; Prokić, L.T.; Racić, G.M.; Pešić, M.B.; Bojović, M.M.; Sierka, E.M.; Kalaji, H.M.; Panković, D.M. Effects of Trichoderma harzianum on photosynthetic characteristics and fruit quality of Tomato plants. Int. J. Mol. Sci. 2021, 22, 6961. [Google Scholar] [CrossRef] [PubMed]
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Figure 1. Root colonization percentage of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. The means followed by the same letter do not differ significantly at α = 0.05.
Figure 1. Root colonization percentage of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. The means followed by the same letter do not differ significantly at α = 0.05.
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Figure 2. Earliness of flowering of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. The means followed by the same letter do not differ significantly at α = 0.05.
Figure 2. Earliness of flowering of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. The means followed by the same letter do not differ significantly at α = 0.05.
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Figure 3. Effect of Trichoderma spp. on flowering, nutritional status, and content of chloroplast pigments in leaves of Gladiolus hybridus.
Figure 3. Effect of Trichoderma spp. on flowering, nutritional status, and content of chloroplast pigments in leaves of Gladiolus hybridus.
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Figure 4. Flowering of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. (A) Length of inflorescence shot (cm); (B) length of inflorescence (cm); (C) number of flowers in the inflorescence; (D) diameter of flower (cm). The means followed by the same letter do not differ significantly at α = 0.05.
Figure 4. Flowering of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. (A) Length of inflorescence shot (cm); (B) length of inflorescence (cm); (C) number of flowers in the inflorescence; (D) diameter of flower (cm). The means followed by the same letter do not differ significantly at α = 0.05.
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Figure 5. Chloroplast pigments content (mg·g−1 FW) in leaves of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. (A) Chlorophyll a + b; (B) carotenoids. The means followed by the same letter do not differ significantly at α = 0.05. FW, fresh weight.
Figure 5. Chloroplast pigments content (mg·g−1 FW) in leaves of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. (A) Chlorophyll a + b; (B) carotenoids. The means followed by the same letter do not differ significantly at α = 0.05. FW, fresh weight.
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Table
Table 1. Content of macroelements (% in DW) in leaves of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. The means followed by the same letter for column-wise and row-wise do not differ significantly at α = 0.05. DW, dry weight.
Table 1. Content of macroelements (% in DW) in leaves of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. The means followed by the same letter for column-wise and row-wise do not differ significantly at α = 0.05. DW, dry weight.
YearTrichoderma spp.Mean
NoYes
% in DW
N
20182.62 a2.51 a2.56 a
20192.42 a2.49 a2.45 a
Mean2.52 a2.50 a
P
20180.51 a0.79 b2.56 b
20190.47 a0.81 b2.45 b
Mean0.49 a0.80 c
K
20184.92 a6.52 b5.72 b
20194.76 a7.34 c6.05 b
Mean4.84 a6.93 c
Mg
20180.21 a0.18 a0.19 a
20190.15 a0.16 a0.15 a
Mean0.18 a0.17 a
Ca
20181.20 a1.36 b1.28 b
20191.18 a1.42 c1.30 b
Mean1.19 a1.39 c
Table
Table 2. Content of microelements (mg kg−1 in DW) in leaves of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. The means followed by the same letter for column-wise and row-wise do not differ significantly at α = 0.05.
Table 2. Content of microelements (mg kg−1 in DW) in leaves of Gladiolus hybridus ‘Advances Red’ after application of Trichoderma spp. The means followed by the same letter for column-wise and row-wise do not differ significantly at α = 0.05.
YearTrichoderma spp.Mean
NoYes
mgˑkg−1 in DW
Mn
2018124.4 a125.0 a124.7 a
2019127.2 a126.2 a126.7 a
Mean125.8 a125. a
Cu
201810.8 a11.0 a10.9 a
201911.2 a11.2 a11.2 a
Mean11.0 a11.1 a
Zn
201820.2 a40.4 b30.3 b
201918.6 a42.2 b30.4 b
Mean19.4 a41.3 c
Fe
2018200.0 a262.0 b231.0 b
2019202.2 a258.4 b230.3 b
Mean201.1 a260.2 c
B
201852.2 a60.4 b56.3 b
201954.6 a61.6 b58.1 b
Mean53.4 a61.0 b
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Andrzejak, R.; Janowska, B. Flowering, Nutritional Status, and Content of Chloroplast Pigments in Leaves of Gladiolus hybridus L. ‘Advances Red’ after Application of Trichoderma spp. Sustainability 2022, 14, 4576. https://0-doi-org.brum.beds.ac.uk/10.3390/su14084576

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Andrzejak R, Janowska B. Flowering, Nutritional Status, and Content of Chloroplast Pigments in Leaves of Gladiolus hybridus L. ‘Advances Red’ after Application of Trichoderma spp. Sustainability. 2022; 14(8):4576. https://0-doi-org.brum.beds.ac.uk/10.3390/su14084576

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Andrzejak, Roman, and Beata Janowska. 2022. "Flowering, Nutritional Status, and Content of Chloroplast Pigments in Leaves of Gladiolus hybridus L. ‘Advances Red’ after Application of Trichoderma spp." Sustainability 14, no. 8: 4576. https://0-doi-org.brum.beds.ac.uk/10.3390/su14084576

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