Effect of the cereal-legume intercropping on Phosphatases and Phytase activity under alkaline soil.

doi:10.21203/rs.3.rs-3627440/v1

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Effect of the cereal-legume intercropping on Phosphatases and Phytase activity under alkaline soil.

https://doi.org/10.21203/rs.3.rs-3627440/v1

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The purpose of the study is to evaluate the effectiveness of intercropping systems cereals (Durum/hard Wheat)-legume (Cicer arietinum) on phosphorus (P) acquisition, pH soil variation, and the variation in enzymatic activity, through root-induced processes in semi-arid soil of South Tunisia. Split plot experiments with triplicate repetitions were carried out in southern Tunisia during two years of field (2020–2021 and 2021–2022). These comprise mono-crop chickpea (CK) and mono-crop durum wheat/ wheat durum (DW/WD), durum wheat intercropping (DW-C), and chickpea intercropping (CK-C). At the complete vegetation stage of durum wheat and chickpea, three soil samples were carried in layer surface for each experimental plot. For the analyses of soil, the P total, Olsen-P, phytase, acid phosphatases, and pH were carried out in the experiment. The obtained findings show a significant amelioration in P total contents in DW-C by 28% and 26% to DW, and 94% and 93% than BS during the two years of field experiment (2020–2021 and 2021–2022) respectively. Furthermore, the Study reported an increase of Olsen-P in the rhizosphere of DW-C by around 5%, 42% than DW, and 36%, 65% to bulk soil (BS) during the two-year experiment. Likewise, these results revealed an increase in A-Phase rates in the DW-C rhizosphere during the two agricultural seasons (2020–2021 and 2021–2021), of approximately 26%, 8% than DW and 33%, 67% than BS respectively. As well as the phytase activity indicated an increase in the DW-C rhizosphere by 67% and 69% than in BS and only by 8% and 7% than in DW for the two seasons (2020–2021 and 2021–2021). Indeed, the rhizosphere acidification of rhizosphere was found very much high in CK-C (0.63 pH units and 0.55 units lower than in the BS).

Intercropping

durum wheat

chickpea

phosphorus

A-Phases

phytase

Phosphorus (P) plays a vital role in cells as well as in structural and energetic elements, which is involved in many biological processes, such as is used commonly for growth, production of energy, reaction with redox, fixation of symbiotic nitrogen, and metabolism of carbohydrates (George et al., 2012; Lazali et al., 2020). The result has shown that the synthesis of nucleic acids (DNA, RNA) and components of phospholipids controls stability and cell membrane properties (George et al., 2012; Lazali et al., 2020). It is involved in many other processes of key physiology and biology during growth and plant development. However, P-deficiency in soils around the world often limits plant growth and represents a significant obstacle to increasing agricultural production to meet global food demand (Duchene et al., 2017); which contributes to the excessive use of mineral fertilizers to improve soil and crop quality, but this intensive pratique of fertilizers especially the phosphate fertilization leads to many environmental and socio-economic problems such as pollution of aquatic ecosystems, and consumption of the main resource used for the synthesis of phosphatic fertilizers and considered to be the main responsible of the eutrophication of coastal ecosystems (Jia et al., 2020). Confronted with soil degradation and pollution, researchers need selection strategies to exploit resources of soil for the management of agroecosystems to optimize the nutrient acquisition of crops (Zhang et al., 2021). Regarding cultivation techniques, it is important to highlight the key elements for the development of plant yields, such currently require stricter management, which is based on the strategy of adapting inputs to meet the needs of the crop during its various stages of development. One of the proposed solutions is the introduction of a specific diversity within agroecosystems through the development of intercropping crops (Lalati et al., 2014; Zhang et al., 2021). For millennia, Asians, Americans, and Africans former attracted attention by intercropping to produce premium yields at the lowest inputs and its land-sparing potential (Li et al., 2020 a, b). Intercropping systems are widely used around the world, including maize-wheat (Mupangwa et al., 2021), bean fava maize (Xu et al., 2018), wheat-pea, and maize-soybean (Fu et al., 2019). Under unfavorable conditions, growing legumes and durum wheat have advantages for P uptake (Cheng et al., 2014; Gong et al., 2020). Additionally, legume is gaining attention for their impact on the sustainability of agriculture and their nutritional and health benefits. Also, it has been found in tropical and Mediterranean regions, that there is a low availability of phosphorus in the soil which is an important nutritional factor for limiting the production of legumes (Latati et al., 2014; Lazuli et al., 2020). Therefore, intercropping can potentially increase crop production and reduce consumption simultaneously (Rezaei-Chiyaned et al., 2021). Better use of resources such as sunlight, water, and nutrients; can help to reduce the need for supplemental irrigation and fertilization which can be expensive and damaging to the environment and nation (Ibrahim et al., 2019; Christie et al., 2020), thus can help to reduce soil erosion increase the yield of crops because of the beneficial effect of crop diversity (Rezaei-Chiyaned et al., 2021). Intercropping of cereals and legumes has been reported to conserve soil and water in some landscapes and provide stable yields (Melkamu, 2023). Smallholder farmers often grow cereal-legume intercrops due to legumes' ability to adapt to disintegrating soils and influence soil health (Begam et al., 2020). Legumes can fix nitrogen from the atmosphere in the soil which increases fertility and reduces the soil nutrients for replenishment (Meena et al., 2015b). Pulses are soil-improving crops with significant soil health benefits that must become an important part of agricultural systems (Dhakal et al., 2016). The soils of semi-arid and arid regions are usually low in P and pose agronomic problems and environmental. Hence, the objectives of the current study were i) to evaluate the impacts of intercropping legume (chickpea)-cereal (durum wheat) on the P availability ii) investigate the variation in enzymatic activity du sol (A-Phase et Phytase) that contribute to P availability iii) and evaluate the potential role of durum wheat–chickpea intercropping in increase biomass, yield and pH variation as compared to sole crops in a low P soil under arid ecosystems of southern Tunisia.

Experimental Site Description

The experiment was conducted over two years (2020–2021 to 2021–2022) in the semi-arid soil of Mednine, South Tunisia (altitude 17m latitude 33°29'56.7'N longitude 10°38'41.50'E), where the climate is arid with an average temperature of 20.3°C. In April, total precipitation reaches a maximum of 4.4 mm. When the sown of crops is done, it reaches the experimental site of soil which is assessed by the top layer of conventional sampling (0–30). The soil is made up of around 79% sand, 5% loam and 11% clay. At 25°C, the soil has an alkaline pH and an electrical conductivity of 1.74mS/cm at 25°, indicating that it is unsalted. According to the Baize classification (1988), organic matter is relatively low (0.8%) and the total limestone concentration in the soil is relatively high (18.7%) with a low active limestone content (2.4%).

A Cropping system and Plot filed design

Simeto durum/hard wheat (Turgidum Triticum durum), Amdoun1 chickpea (Arietinum cicer.), and their intercropping method were included in the study. The test is performed in full field with three repetitions of a complete random block experimental device. The plot is divided into 9 micro-plots each micro-plot shows one growing modality (thus, 3 micro-plots (3 repetitions) per modality): the three cultivation methods are; (i) pure durum wheat (DW), (ii) pure chickpeas (CK), and (iii) chickpeas-durum wheat (DW-C and CK-C). Each micro-plot is 3 m by 1.5 m. The spacing between two micro-plots is 1 m horizontally and 1 m vertically. To lessen the edge effect, a 1 m border is left all around the plot vertically.

Plant and soil samples:

Soil and plant samples were taken at the chickpea and durum wheat flowering stage. It is therefore appropriate to assess the dynamics of nutrients in the soil during these times since they are marked by considerable biological activity of the soil and maximum nutrient absorption. During the flowering period, which is 90 to 100 days following sowing, samples of the rhizosphere of the soil as well as plant biomasses (aerial and root portion) were obtained for various soil and plant tests. To determine the physical characteristics, the soil was dried free-ranging, ground, and sieved to a thickness of 2 mm. Properties are determined by using a standard protocol like Robinson's method of particle size by pipette. Thus, a pH meter will be used to determine the pH hydrogenated potential. Using a conductivity meter determines the electrical connection of the CE. Bernard's altimeter is used to calculate the total limestone using the geometrical method. The Walkley and Black method is used to calculate organic carbon levels. The following formula is used to create biological matter: MO (%) = C% *1.72. using the Kjedhal method, the total nitrogen was calculated. Total phosphorus is determined by nitric acid digestion and assailable phosphorus is evaluated by the Olsen method. The flame photometer was used to measure the amounts of Ca, Mg, K, and Na in the soil. Plant biomass was cleaned before being 48 hours of 60°C drying. Biomass components (aerial and root) were then weighed and set aside for total phosphorus and nitrogen.

Statistical analyses

Analyses of one-way variance (ANOVA) is a 0.05 probability factor conducted on the P total of the cropping system, Olsen-P, phytase, and pH in the rhizosphere; as well as the concentration of root and soot found in phosphorus. A multiple probability of 0.05 has shown the large gap differences in which the values were determined by Tukey’s multiple test. The analysis of statistics was carried out by the usage of XLSTAT.

Availability of rhizosphere in P

There is a change in P total in every season of flowering. However, combined crops of chickpeas and wheat durum have found a greater content for P. About 94% has shown to rise to BS while 28% rise shown in the initial season of DW. The total rise in P total for chickpeas was observed to be greater for CK-C. The only there is one difference that has been found is that there is a difference of 3% for CK. In the next season, it has been observed to increase by 26% in DW-C compared to DW while a 1% drop has been observed at about 93% than to BS. In addition, only a 0.05 difference was significantly noticed because only a 4% difference has been recorded to DW. Combining crops (Wheat durum and chickpeas) has been compared with monoculture and soil in bulk has profoundly appeared to have higher concentration for Olsen-P, which has been increased and has shown greater DW and DW-C, merely found a difference of 36% and for DW-C found 65% which is opposed to BS. If chickpeas merge win merge in a huge quantity of soil, along with the quantity of Olsen-P added, it can be observed the difference is 16% and 4% for CK-C whereas, For CK, the result will appear like 49% and higher percentage about 64% for BS, which found to be a significant difference of 0.05.

Ranges in enzymatic activities

In each season, the content of A-phase chickpeas and durum wheat in the rhizosphere crossed from various experiments. Indications in data were found to be a significant difference of 0.05 which has posed an impact on A-phase content. DW-C was observed to grow in the initial season by 33% and 26% which is considered a higher line for BS and DW. It can observed that the A-Phase is merely about 37% higher than in the CK-C while 6% was seen as higher in CK. These results have found that the DW-C increase after the first season of agriculture was 22% higher for DW and 34% higher than BS during the year 2021–2022. Moreover, the result finding has found that chickpeas crops (CK-C) have resulted in medium growth of 6% in comparison to BS and about 38% to the rhizosphere of CK. The coming crop of Chickpeas and durum wheat has profound upper levels for A-Phtyase to elevate the activity in the rhizosphere of 0.05. The increase in bulk soil and monoculture activity of rhizosphere was found to be high for DW-C which is about 67% and for BS 69%, and for DW, it only rose by 8%, and for BS only 7% rise during both seasons for agriculture processes. Hence, the significant difference for P is less than 0.05, because the percentage of DW-C is only increased by 67% and 69% for BS, and for the case of DW, it is only for 8% and 7%. Merging in bulk soil has also found a higher percentage about 65% for CK-C and 66% than in BS.

Variation in pH

The results of rhizosphere chickpeas in intercropping and solitary cropping have significantly shown lower results for pH. Acidification of the rhizosphere is found to be a core factor for the initial seasoning growth for CK-C has found a low pH value, which is 0.63, and can be said that it is lower compared to BS. The pH value in the soil is decreased for CK to BS, roughly about 0.56 units. In contrast, the results have shown the contrast result in the next season after the first season it has about a pH value of 0.55 units but is still lower than the BS for CK-C. the value is likely to drop again by roughly 0.57 units during the 2021–2022.

Plant biomass and P content

During the stage of flowering, the content of the P plant while intercropping to root and shoot poses a serious cause which creates an effect on durum wheat and chickpeas. In the comparison of CK-C with pure chickpea (CK), the root and shoot for biomass rose by 17% and 26%. Whereas, biomass root and shoot rose by 26% and 37% for DW-C. While in the next trial after the 1st trial season, found high DW compared to that of pure chickpea (CK), together the biomass root and shoot found to be rise by 4% and 11% for CK-C. Moreover, the Concentration of phosphorus in the part of root and shoot appeared to be higher for the durum wheat in the content of P which is found in the combined crop of durum wheat and for the chickpea. In the initial growth period, it increases by 13% in the portion of root and shoot of DW-C and is 3% higher than DW. It has also been found that chickpea accounts for 23% of adsorption about 6% of adsorption for CK in the content of root and shoot. When comparing the DW, adsorption of P in the parts of the root and shoot in DW-C grow in the secondary trial season accounts for 16% and 5%. In addition, adsorption rates were observed to be low compared to 24% and 4% for CK-C in the parts of the root and shoot as a significant difference was found at 0.05.

Increased availability of P in the rhizosphere

In many experiments, the P level of concentration has consistently seen be increase. It has been reported from past research studies that P-intercropped legumes are available and cereals crops of the rhizosphere (Latati et al., 2016). This study also reported the availability of P (Olsen-P) in the content of legumes and intercropped cereal crops. Moreover, the study also suggested that phosphorus deficiency can promote the availability of root induction process in soil alkaline. Acidification in the rhizosphere legumes exerts the nodules of phosphatase carboxylate through the indirect activity of microbial (Latati et al., 2014). The acidity of the legumes used for rising purposes can be accessible to the cereal crop of the rhizosphere in the intercropped phosphorus. Existing acidity in the phosphorus deficiency leads to a rise in the availability of P in the content of soil and can lead to a rise in the number of common beans in the process of legumes (Latati et al., 2014). It might be interlinked with the rhizosphere of cowpeas in an acidic situation. A rise in the content of P is linked with the biomass plant for the durum wheat to be intercropped. Moreover, phosphorus availability has found that there might be a positive correlation between the chickpeas. The study also reported that growth improvement improves the nutrients in the P content as well (Latati et al., 2014). Research finding also shows that P mineralization can promote the chickpeas and the minerals being absorbed by the maize intercropping, which is the cause of phosphate activity that does not find any facilitation for the chickpeas to grow (Latati et al., 2016).

The variation in enzymatic activity

An important role played by the acid phosphatase in the content of P for remobilization transport and production (Lazali et al., 2020). The content of P can be found as an organic complex such as phosphonates, protein, lipids, inositol, and sugar in addition to inorganic ones. Biomass cellular in the content of the soil is decomposing. A few enzymes such as Phytases and Phosphatases breakdown aid in the chemical compound (Pandey et al., 2017). Acid phosphatases and alkaline phosphatases are classified as non-specific by nature in real (Lazali et al., 2021). Acid phosphatases (ATPase) are found commonly in every organism as they are primarily responsible for organophosphate mineralization. ATPase activities elevated and suggested that the enzymatic physiological key role to control the activities for nitrogen which are interlinked to stat p nodal, and hence it provides new insights into how legumes regulate the fixation of N2 (Lazali et al., 2018). It may be crucial for ATPase to uptake and then take it to the recycling process of phosphorus in plants, as it helps to maintain the homeostasis of phosphorus in the nodules of roots. Faba soybean in the soil and accumulation of dry matter increased the long-term application of molybdenum uptake in the rhizosphere and the non-rhizosphere. Enzymatic activity of ATPase and microbial phosphorus biomass and gene transcript express to keep secrets to overcome the bioavailability of phosphorus (Rana et al., 2020). According to a report, ATPase is found to be the formation of a special family of A-phases that hydrolyze the phytic acid, which makes the main forms of P reserves in plants (Lazali et al., 2020). So, there could be a huge number of soil to be produced by these enzymes in the soil such as plants, bacteria, and, fungi (Lazali et al., 2017). It catalyzes the reaction based on the mechanism, similarities, and amino acids along with the four families of A-phytases, such as purple acid phosphatase (PAP), helical phytase (HPP), cysteine phytase (CPhy), histidine acid phosphate (HAP) to hydrolyzed (Lazali et al., 2020).

Rhizosphere pH

The pH level of rhizosphere soil can be altered by legumes, hence increasing the availability of cereals nutrients. The soil becomes acidic when beans are grown under acidic pressure conditions due to the release of protons from the root plants accumulate organic anions return to the soil and decompose (Yan et al., 1996, Melkamu, 2023). When beans are grown in phosphorus-deficient regions some plants such as feba beans can form melic and citric acids to acidify their rhizosphere thereby greatly reducing the pH of the growing medium (Weidenhamer et Callaway., 2010; Melkamu., 2023). Legumes form more nitrogen from the air as diatomic nitrogen than from the soil as NO3 which lowers soil pH. One of the most common plant responses to N2-dependent P is that many plants have formed several duplicate mechanisms that merely deal with the insufficiency of P as reduced soil availability (Hinsinger et al. 2003; Lazali et al., 2021). Beans of the rhizosphere have demonstrated the proton extrusion H + which is linked to the dimensions of acidifying higher roots (Alkama, Ounane, and Drevon., 2012; Lazali et al., 2021). An increase in acidification of the rhizosphere can grow under a sustained low supply of phosphorus of vulgaris (Kouas et al., 2009; Lazali et al., 2021). Because of the P shortage, nodular plants are more likely to assimilate extra cations than anions in the following situations. (Tang et al., 2009) The rhizosphere's H + net outflow was significantly increased. As a result, phosphorus deprivation affects the balance of cations and anion absorption, which in turn affects the release of H⁺ at the root-soil interface to balance the positive charge and control the cytoplasmic pH level of root cells. The rise in rhizosphere acidity suggested that the tuberculum root's acidification had a role in Medicago Truncatula's response to phosphate deficit. Particularly in soil, this feature can modify the interaction of rhizospheres and more often it promotes the uptake of Pi at the interface of soil and root. Soils that contain phosphorus and calcium include calcareous, natural soils, along with acidic soils of phosphorus fertilizers (Zhang et al., 2014). There are certain plant species like chickpeas, white lupine, soybeans, and lentils that release the substances of phosphorus-mobilizing as organic acid. For exemple oxalate, citrate, malate, phosphatases, and protons (Lopez-Arredondo et al., 2014; Lazali et al., 2021). Legumes are found to have a higher capacity to let out carboxylate than any other species of plant due to their bushy roots (Lambeth et al., 2013). Other species of chickpeas with phosphorus deficiency also increase organic anion efflux from roots (Wouterlood et al., 2004), brood beans, alfalfa, as well peas (Maltais-Landry, 2015; Wang et al., 2020).

Plant biomass and P content

The increased availability of P in the rhizosphere may be the cause of the rise in plant biomass. Intercropping with leguminous crops has reportedly resulted in higher grain yields: Cowpeas or broad beans for maize (Dahmardeh et al., 2010) and wheat durum (Latati et al., 2014). Process of intercropping cereals with beans crops adding the phosphorus while food supplement. To compare the chickpeas that are grown in isolation creates competition between chickpeas and wheat to intercrops which pose a significant negative impact on the growth of chickpeas. Moreover, the competition of nutrients between the selective crops might be noticed by the depth of root difference the extension of root lateral, and the density of root (Khanal et al., 2021). A study has reported the uptake of phosphorus wheat found to be higher when it was in conjunction with the chickpeas than when it was grown in isolation in that specialty (Latati et al., 2016), as it has reported that comparable outcomes have been observed in the grown in isolation for maize mixed and crops of bean. Organic phosphorus can be hydrolyzed by secreting phosphorus because the wheat content rises in the phosphorus, which has been turned to enhance the uptake of phosphorus for wheat. The consistent results are found to be higher compared to the uptake of phosphates in the consumption of wheat which has been grown below the circumstance conditions of the field at the isolated places compared to the case of chickpeas where the uptake of Phosphorus. When planting in mixed conditions, bean plants in mixed corn conditions have increased their phosphorus content in the wheat. This might occur because of the chickpea’s capacity to hydrolyze the phosphorus organically by secreting the phosphorus, thus promoting the absorption of phosphorus in the quantity of wheat (Latati et al., 2016).

The study concludes the overall process of intercropping, which improves the fertility of soil management and all the physical characteristics of chemicals in the soil. Durum wheat and Chickpeas cultivated on the lower side of P soils in such circumstances for two seasons of agriculture as to address the core subject for the separate and inter-collective crop. Outcomes tests have revealed that there are substantial variations in the biomass plant and the uptake of P plants. In addition, the quantity of A-phase and Phytase were found das higher for chickpea and the durum wheat in comparison to the classification of monoculture. These findings demonstrate the availability of P in the durum wheat which is solely based on PH value. More often, it can be seen that durum wheat for intercropping rhizosphere becomes more alkaline while the intercropping for chickpeas rhizosphere is observed as a more acidic formation. Lastly, the current study concludes that to support the advantages of intercrops from yielding grains for P nutrition for both legumes and cereal crops and then increasing agroecosystem sustainability.

Author Contribution

- Amira Souidand Wissem Hamdi wrote the main manuscript text.-Amal Attallah participate in the collection of experimental data!-Mohamed Farissi, Boulbaba L’taief and Mohamed Faouazi Zagrarni reviewed the manuscript

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Table 1: Physico-chemical results after soil characterization

Parametres	Results
pH	7.577
CE (mS/cm)	1.74
Total nitrogen (mg/g)	0.040
P total (mg P/kg)	0.661
Olsen-P (mg P/kg) A-Phase (µmol PNP/g) Phytase (mgP /kg)	0.895 11.41 2.22
Potassium (ppm) Sodium (ppm)	83.143 71.183
Total limestone (%)	18.741
Active limestone (%)	2.460
Carbon (%)	0.471
Organic material (%)	0.8

No competing interests reported.

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Effect of the cereal-legume intercropping on Phosphatases and Phytase activity under alkaline soil.

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Abstract

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Introduction

Methods and Equipment

Experimental Site Description

A Cropping system and Plot filed design

Plant and soil samples:

Statistical analyses

Results

Availability of rhizosphere in P

Ranges in enzymatic activities

Variation in pH

Plant biomass and P content

Discussion

Increased availability of P in the rhizosphere

The variation in enzymatic activity

Rhizosphere pH

Plant biomass and P content

Conclusion

Declarations

Author Contribution

References

Table

Additional Declarations

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