Grasp of wheat leaf rust through plant leaves extract and bioagent as an eco-friendly measure

1 Introduction

Leaf rust caused by the pathogen Puccinia has more potential to threaten the yield of wheat by up to 10% as compared to the other two types of rust stem and yellow rust. The severity of the disease depends upon the developmental stage of the plant and its susceptibility of the plant (Duplessis, et al., 2021). The variability of wheat growing conditions favorable climatic patterns, and genomic polyploidy nature it becomes the most financial and nutritional crop (Hassan, et al., 2022). The FAO report on the annual global supply and demand for wheat is ample for wheat production (FAO). The population development rate is increasing, which is estimated at 9 billion by 2025. To meet the demand for staple food the production of wheat is an important interest for its significance turn out (Schnurbusch, 2019).

The favourable condition for rust to infect the plant at high relative humidity with a temperature range of 15–22 °C with optimum temperature for spore’s development is 20 °C in only 6–8 h for the development of disease. Leaf rust infections can cause yield losses beyond 50 percent at even earlier stages. The damage to rust in leaves sometimes is less spectacular than that produced by strip (yellow), but as leaf rust frequently occurs the total yearly losses worldwide are likely more than the others. (Huerta-Espino et al., 2011).

Rust challenges the yield of the wheat crop so chemicals are used in terms to prevent and treat the disease incidence, but the chemical nature of the synthetic fungicide has a worse effect on the health and metabolic life of humans and animals. It also makes wheat single-quality varieties resistant to the pathogen (Hassan, et al., 2022). Some measures are proving very helpful to overcome the losses caused by Leaf rust followed by researchers as well as field workers including cultivars susceptibility, variety genetic resistance, climatic interaction, microorganism disease transmission capacity, and field management process (Temesgen, 2015). Some plants have pathogens-toxic substances that are isolated from plants and administered to diseased plants.

The compounds are referred to as botanical or botanical pesticides. Plant extracts including neem (Azadirachta indica, A. juss), moringa (Moringa olifera), and bioagent Trichoderma herzianum are commonly utilized in botanical substances. The biologist is interested to control the leaf disease severity by non-chemical method therefore researcher's main concern is to elevate the effect of synthetic fungicide causing environmental and biological pollution by bioagents and eco-friendly plant extracts. A plant-like Neem Azadirachta indica, Moringa, has an inhibitory effect on leaf rust (Shabana et al., 2016). The bioagent used as an antagonistic organism like Trichoderma species have been used as biological control agents for over 70 years, but just now have strains been widely available. Many strains of Trichoderma have been found to have potential use under biological control, especially T. harzianum, T. viride, and T. virens (Monte and Liobell, 2003).

These biocontrol agents are known to be natural having no side effects on human life but having a clear and effective result on the targeted organism. The bioagent is safe to use against diseases like wheat leaf rust due to its eco-friendly nature, cheapest, and ease to access. The research study is designed to assess the vulnerability of commercial wheat varieties against leaf rust caused by Puccinia triticiana and its biocontrol by the natural bioagents Trichoderma and plant leaf extracts. It may be a concern for future to work on the active substance of the used biocontrol agents to cope up the economic loss of wheat.

2 Material and method

Plant extracts of two plant species, including Neem leaves (Azadirachta indica), Moringa (Moringa oleifera) and Bioagent, (Trichoderma harzianum), were tested in the field conditions of regional Agriculture Research Institutes to determine their effectiveness as an inducer of materials to resist the infection of leaf rust. Fresh leaves of Neem and Moringa were taken from the field. Plant samples were cleaned to clear dust with tap water. Three different concentrations of 50 ml, 100 ml, and 150 ml of plant leave extract were made. For this purpose, take 50 g, 100 g, and 150 g of fresh leaves in pre-chilled mortar and pistil in chilled distilled water. All the plant leaf extracts were filtered with the help of a muslin cloth. The concentrated filtrates were diluted with distilled water to make a final stock solution of 50 ml, 100 ml, and 150 ml, respectively. For Trichoderma herzianum application the culture of Trichoderma herzianum was obtained from the Plant Pathology Department, Regional Agricultural Research Institute, Bahawalpur. The spore suspension of Trichoderma harzianum is prepared by adding 10 g of Trichoderma herzianum in 1 L of water with 50 ml of Limonoil. The Limonoil was used for its adhesive and wetting property that make the spore suspension more effective (Seng, et al., 2014). The stock solution is then stored at 4 °C.

For the experimental trial Morocco (Susceptible check) was sown in a single row of 2 m length with 30 cm row to row spacing to determine the efficacy of treatment in different concentrations. The inoculation of uredospore of Puccinia triticiana to wheat plant at two stages mainly at the seedling and booting stage (Shabana et al., 2017). The dusting method was used in the volume of 20 g talcum powder and 1 g of Puccinia triticiana spores at high temperature and humidity. Before dew began, dusting was performed at sundown. Inoculation was performed at the 7–8th stage of development. Distilled water was sprayed on the untreated controls The disease evaluation was recorded after observation based on the average coefficient of infection (ACI). The foliar spray was applied to the disease check-in in Morocco.

3 Biochemical analysis

4 Result and discussion

4.3

4.3 Biochemical analysis

4.3.1

4.3.1 Chlorophyll content

The chlorophyll content shows that all the treatment shows an active response in increasing the total chlorophyll of the treated plant as compared to the control.

Trichoderma herzianum has significant interaction with the biochemical product of plants by increasing the photosynthetic activity of the plant. The photosynthesis activity increased by 0.45, 0.49, and 0.52 at 150 ml conc. of Moringa, 150 ml Neem, and Trichoderma herzianum respectively (Table 4). On the other hand, the Neem at 50 ml concentration shows minimum chlorophyll content as compared with the control (Fig. 1).

Table 4 Efficacy of plant extracts and bioagent Trichoderma herzianum on chlorophyll content.

Neem:	Sample dry weight	Total chlorophyll content
(Conc. 50 ml)	0.25 g	0.31
(Conc. 100 ml)	0.25 g	0.4
(Conc. 150 ml)	0.25 g	0.49
Moringa:
(Conc. 50 ml)	0.25 g	0.43
(Conc. 100 ml)	0.25 g	0.46
(Conc. 150 ml)	0.25 g	0.45
Trichoderma Harzianum	0.25 g	0.52
Control	0.25 g	0.3

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Fig. 1

Efficacy of plant extracts and bioagent Trichoderma herzianum on chlorophyll.

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4.3.2

4.3.2 Phenolic content

The total phenolic content at 0.25 g dry weight of wheat leaf. There is a variation in results showing at different conc. Of plant extract and plant bio stimulant Trichoderma herzianum. All the bio-agents show positive results while Neem 50 ml concentration shows a minimum increase or less than the control. The increase of total phenolic content by Moringa 150 ml concentration, Neem 150 ml concentration, and Trichoderma herzianum by 3.56 mg, 7.3 mg, and 8.1 mg, respectively (Table 5, Fig. 2).

Table 5 Efficacy of plant extracts and bioagent Trichoderma herzianum on phenolic content.

Neem	Sample dry weight	Total phenolic content
(Conc.50 ml)	0.25 g	0.81 mg
(Conc. 100 ml)	0.25 g	1.37 mg
(Conc. 150 ml)	0.25 g	7.3 mg
Moringa
(Conc.50 ml)	0.25 g	1.08 mg
(Conc. 100 ml)	0.25 g	1.22 mg
(Conc. 150 ml)	0.25 g	3.56 mg
Trichoderma harzianum	0.25 g	8.1 mg
Control	0.25 g	0.92 mg

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Fig. 2

Efficacy of plant extracts and bioagent Trichoderma herzianum on phenolic content.

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5 Discussion

To control rust disease the scientist, urge many managements of chemical, genetic, farming practices, and biological control (Harish et al., 2008). For eco-friendly measures, the earth reverses its system to a natural system. The application of plant products and bioremediation agents for plant protection against disease has recently become more important due to their widely accessible nature, bioagents, microorganism as a biostimulant, antibacterial activity, simple biodegradability, phytotoxicity, and generating host resistance. Numerous researchers have previously shown that certain plants have antifungal action against several plant diseases like leaf rust. Many bacterial and fungal strains are useful to control the disease. The fundamental processes of disease control by plant parts extracts are not apparent, although the participation of inducing resistance is considered (Shabana et al., 2017).

Several publications (El-Sharkawy et al., 2018) reveal that extracts of plants are efficient biocontrol agents against a variety of diseases in plants. Plants are capable of synthesis of secondary aromatic metabolites such as quinones, phenols, phenol acids, tannins, flavones, flavonoids and coumarins (Han et al., 2018). The phenolic structural components, such as eugenol, carvacrol and thymol, have been very potent against infections.

These groupings exhibit antibacterial action and act as strategies for plant protection against harmful microorganisms (Al-Jumaili, et al., 2018). Meliaceae plants, in particular neem, have at least 35 physiologically active components, the most important of which are combine and azadirachtin (Sujarwo et al., 2016).

The Neem plant extract shows a compatible response against leaf rust with about 98.99% efficacy when applied at a mature stage of wheat under field conditions in one and two sprays at the 4-day interval. While in seed soaking with Neem extract and at the seedling stage foliar application reduces leaf rust infection (ACI) and increases the efficacy of plant extract in controlling the disease. The use of Neem plant leaf extracts is also effective in increasing the 1000 kernel weight, spike length and plant height (Shabana et al., 2017).

Trichoderma herzianum is a beneficial microorganism used as an antagonist to many microorganisms’ diseases like Leaf rust. It is also used to enhance the fertility of the soil, as a biopesticide, and as a biostimulant. These bioagents are non-polluting, low-cost, and non-hazardous, and may be made with readily available processes and materials. A second response in the improvement of the host defensive mechanism can be the type of action of abiotic inducers to prevent plant disease.

The bioagents Trichoderma harzianum was used to address the suppression of disease severity for three types of disease including stem rust, powdery mildew, and leaf blotch in different sowing dates trial (El-Mougy et al., 2020). The bioagent showed reduction in disease as compared to the control group. Infield experiment the Trichoderma herzianum increase root development and growth by the absorption of more nutrient from the soil. As compared to the control group the treatment with Trichoderma herzianum showed efficacy in 1000 kernel grain weight, spike length, a number of rootlets and tiller number also increase.

The rise in wheat production was comparable to the studies of Sallam Nashwa et al. (2008), who discovered that the preparation of Trichoderma spp. treatments increased the vegetative yield of bean plants when contrasted to infected controls.

6 Conclusion

The organic control of rust using plant extracts is an advanced alternative rust management technique that is current, advanced, and risk-free. Two plant leaf extracts at different concentrations and bioagent Trichoderma herzianum were tested to check leaf rust disease of the wheat (Neem, Moringa with 150 ml, 100 ml and 50 ml concentration). In the sowing stage, Neem extract reported a spore germination inhibition of 98.99% and a mature wheat plant of 82.90%. Spray on Wheat seedlings treatment with the plant extracts reduced the number of pustules/leaves and increased the effectiveness of foliar spraying. Neem extract was the most efficient treatment in this respect.

When administered one or four days after inoculation, it offered 82.90% control of the illness. After 22 days of inoculation, pustules started to emerge on control, but no pustules on neem 150 ml concentration, followed by Trichoderma herzianum, and moringa 150 ml concentration extracts. A foliar spray was the most efficient in decreasing leaf rust (ACI) infection and the neem extract at the mature plant stage. The applied treatment also shows a significant increase in photosynthesis, chlorophyll and grain production. Resistance to leaf rust in Pakistan's wheat cultivars has been shown in current results.