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Interactive Effect of Nano Chitosan and Soil Mulching on Salt Affected Soil Characteristics and Phaseolus vulgaris L. Productivity

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22 December 2023

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26 December 2023

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Abstract
Soil salinity is seen as a major restriction for crop production, and with water scarcity this problem becomes more complicated. Mulching is crucial to salinity dynamics management by decreasing evaporation with improving the soil’s characteristics. Using chitosan as an eco-friendly biostimulant can enhance plant defense genes during different abiotic stresses. Recently, agricultural research has recognized nanoparticles as a pioneer material due to their distinctive physicochemical features. Therefore, a lysimeter experiment was conducted to investigate the interactive effects of mulching (UNM: un-mulched, WPM: white plastic, RSM: rice straw and SDM: sawdust) and chitosan foliar application (Ch0: control, Ch1: 250 mg chitosan L-1, Ch2: 125 mg nano chitosan L-1, and Ch3: 62.5 mg nano chitosan L-1) on the biochemical soil characteristics and common beans productivity under salt affected soil conditions. Organic mulching (RSM and SDM) treatments significantly improved the soil's organic carbon, available nutrient content, and total count of bacteria. WPM treatment lowered soil EC to 6.63 dS m-1 and increased soil water content to 34.13%. The application of Ch3 caused considerable increases in the plant height, shoot dry weight, root dry weight, grain yield, and nutrient content in the seed. The total fungi count in the soil and Na% in the seed was significantly decreased due to chitosan foliar applications. Moreover, the interactive effect of different mulch materials plus foliar by chitosan applications gave a statistically similar seed yield in both seasons. Overall, this study revealed the potential of the mulching treatments and foliar application of nano chitosan in improved biochemical soil characteristics and common bean productivity under saline soil conditions.
Keywords: 
Subject: Environmental and Earth Sciences  -   Soil Science

1. Introduction

Salt stress has a deleterious effect on global crop production [1]. In semi-arid and arid climates, soil salinization can be the result of natural and climate change impacts, and anthropic activities [2,3]. Globally, salt-affected soil covers approximately 1.1* 109 hectares [4]. At local level, in Kafr El-Sheikh Governorate (current study area, North Nile Delta of Egypt), salt affected soil covers approximately 151.05 hectares (56%) of the total cultivated area [5]. Consequently, a variety of factors, such as freshwater shortage, drought, soil texture, seawater intrusion, leaning on wastewater and inefficient drainage networks can lead to the salt accumulation and soil health degradation [6,7,8]. Fertility disruption, decreased microbial activity, and compromised soil structure are the negative effects of soil salinization [9]. Under salt-affected soil conditions, the osmotic potential is higher in soil solution than in plant root cells [10]. Therefore, it is crucial to fairly manage the salinity dynamics by decreasing soil water evaporation, reducing salt accumulation, and improving the soil’s characteristics in order to increase crop productivity in salt-affected soils.
By mulching with various materials, soil water evaporation can be decreased, the amount of soil water available for plants can be increased, and salt buildup can be reduced in the soil [11]. Currently available various mulching materials such as plastic, non-woven, biodegradable plastic, paper films, and organic mulches, such as straw or wood chips, and gravels [12,13]. Plastic mulching had a positive effect on soil physicochemical properties [14,15]. According to Fan et al., [16], straw mulch can beneficially reduce soil water evaporation which encumbers salt accumulation. Also, sawdust mulching decreased soil EC, increased the soil organic matter content, and reserved the soil available nutrients [17]. Different sources of mulches afflicted soil organic carbon content, total nitrogen, soil pH, exchangeable cations, available phosphorous and EC, and base saturation in soil [18]. At the same time, mulching applications encourage soil biological activities [19,20]. Different mulching materials showed an exhibited the highest growth parameters and yield of common beans (Phaseolus vulgaris L.) under saline stress [21,22,23].
Chitosan (CS; poly _-(1,4)-N-acetyl-D-glucosamine) is non-toxic, non-allergenic, cost-effective, biodegradable, and eco-friendly, and it is used in agriculture as a biostimulant [24]. This natural substance is effective in promoting plant growth and resisting abiotic stress [25,26]. Cataldo et al., [27] found that the chitosan improved several defensive genes in plants, including pathogenic-related genes (glucanase and chitinase). It also reduces the impact of salinity stress on plants and enhances plant growth by regulating cellular osmotic pressure and increasing the availability and uptake of water and essential nutrients [28].
Currently, nanotechnology has recently been extensively utilized in several areas of plant improvement, with nanoparticles (NPs) being replaced by bulk materials [29]. Chitosan nanoparticles (CSNPs) are advantageous due to their interface and surface effects, as well as their small size, which makes them more effective than normal chitosan [30]. The effectiveness of chitosan nanoparticles (CNPs) is enhanced by their small size (less than 100 nm), high aspect ratio, and surface area [31]. Which, their enhancement of plant metabolic activity leads to more efficient transport of active chemicals across cell membranes [32].
Despite using chitosan nanoparticles has been proven to have beneficial impacts on plant productivity, there are limited studies about using chitosan or chitosan nanoparticles to enhance growth and productivity in common bean (Phaseolus vulgaris L.) plants under salinity stress. Also, the interactive effect of nano chitosan and soil mulching on the biochemical characteristics of salt-affected soil and common bean productivity is still unclear. Therefore, this research tried to study the effect of Nano chitosan as a foliar application and organic and inorganic mulching on the biochemical soil characteristics and common bean productivity under salt-affected soil conditions.

2. Materials and Methods

2.1. Experimental Site

A lysimeter experiment was conducted in consecutive two growing seasons during first week of March 2022 and 2023 at the Sakha Agricultural Research Station (30° 56' 53" E, 31° 05' 38" N, with elevation from sea level is about 6 m.) Kafer El-Sheikh Governorate, Egypt. The climate of experimental area is arid climate. Detailed information about the daily rainfall and temperature of two growing seasons 2022 and 2023 (Figure 1) was collected from Weather Station installed at experimental site with an average annual precipitation was 62.21–37.84 mm and temperature were 23.2 and 24.3 °C during cultivation period of 2022 and 2023. The soil characteristics (average values of 48 lysimeter) before sowing indicated that the soil was saline heavy clay (clay 57.97%, silt 25.37%, sand 18.66%, pH 8.16, EC 7.78 dS m-1, ESP 14.76, bulk density 1.42 Mg m-3, soil water content was 27.48%), with a low content of organic carbon 0.620 g g-1, available-N 23.09 mg kg-1, available-K 204.46 mg kg-1, available-P (Olsen method) 11.77 mg kg-1. Also, total count of bacteria was 5.45 x 103 CFU g-1 and total count of fungi was 4.07 x 102 CFU g-1.

2.2. Material

2.2.1. Corp variety

Common bean (Phaseolus vulgaris L.) cv. Giza 6 seeds were obtained from Horticulture Research Dep., Sakha Agri. Res. Station in Kafr El-Sheikh Gov.

2.2.2. Mulching materials

White plastic (30 μm) was obtained from the Arasya Plastic Company, Heliopolis, Cairo Gov. Rice straw, and sawdust were obtained from a farmer and wood machinery in Kafr El-Sheikh Gov.

2.2.3. Chitosan

It was purchased from Chitosan Egypt Company. Chitosan (C6 H11 NO4)n, from shrimp shell, with a deacetylation degree (DD) of about 90-95% and molecular weight: <100 cP.

2.2.3.1. Synthesis and characterizing of nano chitosan

In the present work, the nano-chitosan particles were prepared using the ball milling method for a 60-minute to synthesize nano-sized, after milling, the samples were dried for 10 minutes [33]. Characterizing nano-sized chitosan at Alexandria University was done by scanning electron microscope (SEM), and particle size distribution analysis. A Fourier Transform Infra-Red (FTIR) spectrum is being measured at the Central Laboratory of Tanta University Egypt.

2.2.3.2. Chitosan solution preparation

Different doses of the chitosan or its nanoparticles (250 or 125 and 62.5 mg L-1) solubilized in 800 ml of distilled water with 1% acetic acid. Then, constant stirring until completely dissolved, and then completes a volume to one litter. Finally, the solution was alkalized to pH 6 with 1 M NaOH solution [34].

2.3. Treatment and design

The treatments were arranged as a factorial experiment in a completely randomized design with three replications. There were 2 factors as follows (1) mulching materials, UNM: un-mulched, WPM: white plastic mulching (30 μm), RSM: rice straw mulching (5 cm), and SDM: sawdust mulching (5 cm), (2) the chitosan foliar application, Ch0, Ch1, Ch2 and Ch3: distilled water, 250 mg chitosan L-1, 125 mg nano chitosan L-1, and 62.5 mg nano chitosan L-1 respectively. Each treatment was randomly arranged with 48 lysimeters of 0.64 m2 (80 cm × 80 cm).

2.4. Experiment setup

Two rows were setup in every lysimeter with 60 cm length, 25 cm width and 20 cm height, as the plant spacing was 20 cm. Seeds were sown on 1 March 2022 and 2023. The mulch treatment was applied after 1 week from sowing and foliar application treatments were applied at 15, 30, and 45 days after sowing, the plants received three separate foliar applications. The irrigation water used has a pH of 7.08 and an EC of 2.51 dS m-1 and irrigation was adapted to 60 cm depth through reaching the FC + 5% as a leaching requirement. The fertilization and agricultural practices were done according to common bean cultivation in the North Nile Delta region of Egypt. After full maturity, the plants were harvested on 18 July.

2.3. Sampling and Measurement

2.3.1. Soil Characteristics

Soil samples with a surface area of 30 cm were collected before sowing and after harvesting common beans to be analyzed using the methods cited by [35,36]. Soil microbial communities were determined as outlined by [37].

2.3.2. Yield and quality of seeds:

After 60 days, the plant height (cm) was measured. The shoot dry weight (g), root dry weight (g), and seed yield (kg ha-1) were determined after the full maturity of common beans. For determination of N, P, K and Na percentages in common bean seed. The seed samples were dried, ground, and wet-digested as described by [38]. The N, P, K and Na% were determined according to stander methods [39].

2.4. Data analysis and processing

All data were subjected to analysis of variance (ANOVA) using Minitab v 21. Tukey’s test was performed to make multiple comparisons between different treatments at p≤ 0.05. Data was processed in R (ver. 4.1.3), for Principal Component Analysis (PCA) uses Factoextra packages [40] and Visualize Correlation Matrix using corrplot packages [41].

3. Results

3.1. Characterization of Nano chitosan

Characterizing nano chitosan as shown in Figure (2 a,b,c). Figure (2a) displays the particle size distribution of chitosan nanoparticles that represent an average diameter of 86.4 nm. Figure (2b) displays an SEM image of nano chitosan. The result of SEM was consistent with the DLS measurement in terms of nanoparticle size. The surface structure of nano chitosan powder is smooth, compact, and uniform.
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Figure 2. Characterization of nano chitosan (a) Particle size distribution image (b) Scanning electron (c) microscope (SEM) and FTIR spectrum.
Figure 2. Characterization of nano chitosan (a) Particle size distribution image (b) Scanning electron (c) microscope (SEM) and FTIR spectrum.
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The functional groups of nano chitosan were characterized using Fourier Transform Infrared (FTIR) spectra analysis as shown in Figure (2c).The results indicated that the absorption at wavenumber 3852.39-3449.47 cm-1 showed the presence of O-H. Absorption at wavenumber 2923.05-2855.04 and 2877 cm-1 showed the presence of the methylene and methyl group. Absorption at wavenumber 1634.71 and 1076.53 cm-1, respectively, were related to the C=O stretching and C-O bending vibrations. Absorption at wavenumber 1423.67 cm-1 indicated the C–N stretching which showed the acetyl group. Absorption at wavenumber 1149.52 cm-1 indicated that the vibration absorption CN (NH2) stretching as evidence of amine groups was formed. Absorption at wavenumber 1076.53 cm-1 indicated the CN (H2) stretching because the amine groups were formed. Absorption indicates that intramolecular hydrogen bonds in the structure were very strong.

3.2. Soil parameters

The results of study soil parameters of each treatment were shown in Table 1. In general, the obtained results from the analysis of mulching treatment application, all mulching materials used caused a significant change in all studied soil parameters. Also, chitosan foliar applications, compared with control application (Ch0), using nano chitosan foliar applications caused a slight significantly changes on soil EC, SOM and microbial communities. The interactive effects of different mulching materials and chitosan foliar applications were found significant (at p ≤ 0.05) on all study soil parameters.

3.2.1. Soil Electrical Conductivity

From the analysis of mulching treatments, different mulching materials had different effects on soil EC (Table 1). The white plastic mulch treatment (WPM) was better, which could decrease the soil EC by 7.91 and 12.83% compared to UNM in both seasons, respectively. Ch0 application had the highest soil EC values, averaging 7.42 dS m-1 in 2 seasons (Table 1). Both doses of nano chitosan foliar application (Ch2 and Ch3) showed a slight reduction in soil EC in 2 seasons. Different mulching materials combined with chitosan foliar application reduced EC in soil compared with control (UNM+Ch0). The lowest values of EC (7.04 and 6.51dS m-1) were observed under WPM + Ch3 with insignificant differences with Ch1 and Ch2 in both seasons, respectively (Table 1).

3.2.2. Soil Water Content

The soil water content (SWC) under the mulching treatment increased sequentially compared to UNM (Table 1). AS, WPM and SDM contain relatively a higher soil water content. Using WPM or SDM to cover the soil increased the soil water content by an average by 22.99% or 20.84% compared to UNM in 2 seasons, respectively. Among all the chitosan foliar applications, the soil water content values did not show any significant difference after 2 seasons due to the application of the various chitosan foliar applications (Table 1). There is no significant distinction between the combined soil mulching materials with the chitosan foliar applications in obtained values of SWC (Table 1).

3.2.3. Soil organic carbon

The soil organic carbon was found in the mulching soils significantly larger than un-mulched bare soil (UNM). The SOC content increased by 0.159 and 0.184 g g-1 under SDM treatment, followed by RSM treatment with an increasing 0.097 and 0.130 g g-1 comparing with UNM in 2 seasons, respectively (Table 1). Compared with control application, the nano chitosan foliar application at a rate of 63 mg L-1 (Ch3) had the highest values of soil organic carbon content (0.690 and 0.716 g g-1) in 2 seasons, respectively. Under the organic mulching treatments, SOC content increased with all chitosan foliar applications (Table 1), and the maximum content sawdust mulching with nano chitosan foliar application at a rate of 63 mg L-1 treatment (SDM+Ch3).
Table 1. Soil chemical characteristics under different treatments affect.
Table 1. Soil chemical characteristics under different treatments affect.
Soil parameters EC S.O.C AV. N AV. P AV. K SWC TBC TFC
Seasons 1st 2nd 1st 2nd 1st 2nd 1st 2nd 1st 2nd 1st 2nd 1st 2nd 1st 2nd
Mulching materials
UNM 7.77a 7.60a 0.598d 0.602d 27.26c 27.56c 12.57b 12.38c 201.19d 201.34d 27.14c 27.89b 5.92d 5.96c 3.61a 3.31a
WPM 7.15c 6.63d 0.643c 0.660c 33.61b 34.60b 13.68b 14.19b 256.36c 257.31c 33.54a 34.13a 6.96c 7.23b 3.06ab 2.47b
RSM 7.51b 7.16b 0.695b 0.732b 36.33a 36.90a 16.89a 16.82a 266.13b 268.91b 29.63b 29.16b 7.45b 7.49ab 3.15ab 2.31b
SDM 7.40b 7.01c 0.757a 0.786a 35.50ab 36.17ab 16.10a 15.82a 283.55a 284.63a 32.94a 33.55a 7.98a 8.33a 2.40b 1.33c
F-Value 30.07 116.89 81.08 66.08 58.67 56.51 47.11 50.21 862.68 1156.48 64.99 27.49 43.15 18.26 4.86 13.76
P-Value <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 0.007 <.0001
Chitosan foliar application
Ch0 7.61a 7.23a 0.659b 0.668b 32.32 32.77b 15.30 15.42 251.03 252.05 31.15 31.58 6.64c 6.66c 3.43 2.86a
Ch1 7.47ab 7.12ab 0.669ab 0.691ab 33.35 34.03a 14.74 14.74 251.86 253.11 30.87 31.22 7.10b 7.28b 3.03 2.33b
Ch2 7.41b 7.05b 0.676ab 0.705ab 33.44 34.14a 14.66 14.61 252.09 253.31 30.72 31.08 7.26a 7.45ab 2.92 2.15b
Ch3 7.35b 7.00b 0.690a 0.716a 33.59 34.29a 14.55 14.43 252.26 253.71 30.51 30.86 7.31a 7.62a 2.85 2.08b
F-Value 5.75 7.08 2.88 4.42 1.17 2.11 1.29 2.51 0.20 0.45 0.53 0.26 55.46 57.15 1.31 19.91
P-Value 0.003 0.001 0.051 0.01 0.338 0.043 0.294 0.076 0.894 0.722 0.667 0.856 <.0001 <.0001 0.288 0.002
Interaction
UNM Ch0 7.89a 7.78a 0.588f 0.555g 26.44c 24.39d 12.93de 12.52cd 200.64c 199.79f 27.37e 28.19bcd 5.21h 4.75b 4.07 3.92a
UNM Ch1 7.77ab 7.61ab 0.599f 0.601fg 27.43bc 28.52cd 12.50e 12.41cd 201.28c 201.65f 27.13e 27.87cd 6.09g 6.19ab 3.53 3.28ab
UNM Ch2 7.72abc 7.53abc 0.607f 0.638efg 27.55bc 28.58cd 12.48e 12.32d 201.30c 201.87f 27.06e 27.87cd 6.17g 6.32ab 3.49 3.07ab
UNM Ch3 7.69abc 7.49abcd 0.596ef 0.613fg 27.61bc 28.77bcd 12.38e 12.29d 201.53c 202.03f 26.98e 27.64d 6.21g 6.57ab 3.35 2.98ab
WPM Ch0 7.36bcde 6.75fghi 0.625def 0.632efg 32.87ab 34.20abc 14.17bcde 14.93abcd 255.70b 255.85e 33.90a 34.54a 6.66f 6.80ab 3.38 3.00ab
WPM Ch1 7.16de 6.67ghi 0.638def 0.659def 33.77a 34.64ab 13.63cde 14.10bcd 256.31b 257.48de 33.59ab 34.22ab 6.90ef 7.15ab 3.05 2.42ab
WPM Ch2 7.06de 6.58hi 0.646def 0.665cdef 33.85a 34.73a 13.50de 13.99bcd 256.55b 257.70cde 33.50ab 33.85abcd 7.10de 7.42a 2.92 2.26ab
WPM Ch3 7.04e 6.51i 0.665cdef 0.68bcdef 33.94a 34.85a 13.42de 13.72bcd 256.87b 258.21bcde 33.16abc 33.92abc 7.18de 7.53a 2.89 2.19ab
RSM Ch0 7.66abc 7.31bcde 0.685bcde 0.719abcde 35.39a 36.53a 17.33a 17.73a 264.45b 268.62bcd 30.13abcde 29.64abcd 7.03def 7.09ab 3.48 2.92ab
RSM Ch1 7.53abcd 7.19cde 0.694bcd 0.723abcde 36.47a 36.93a 16.89ab 16.56ab 266.35b 268.76bc 29.81bcde 29.10abcd 7.37cd 7.45a 3.11 2.18ab
RSM Ch2 7.48abcde 7.10def 0.696bcd 0.733abcde 36.62a 37.01a 16.71abc 16.52ab 266.86b 268.92b 29.43cde 29.06abcd 7.65bc 7.64a 3.04 2.09ab
RSM Ch3 7.38bcde 7.05efg 0.704bcd 0.752abcd 36.86a 37.15a 16.63abc 16.44ab 266.86b 269.35b 29.16de 28.85abcd 7.74abc 7.79a 2.97 2.05ab
SDM Ch0 7.52abcde 7.09ef 0.739abc 0.76abc 34.58a 35.97a 16.76ab 16.51ab 283.32a 283.92a 33.19abc 33.94abcd 7.65bc 8.01a 2.78 1.61b
SDM Ch1 7.42abcde 7.03efg 0.743abc 0.778ab 35.72a 36.06a 15.95abcd 15.91ab 283.49a 284.56a 32.96abcd 33.70abcd 8.02ab 8.33a 2.41 1.43b
SDM Ch2 7.36bcde 6.98efg 0.754ab 0.785a 35.75a 36.23a 15.93abcd 15.61ab 283.63a 284.77a 32.87abcd 33.53abcd 8.11a 8.41a 2.23 1.18b
SDM Ch3 7.28cde 6.95efgh 0.793a 0.818a 35.94a 36.41a 15.76abcd 15.26abc 283.78a 285.25a 32.74abcd 33.02abcd 8.13a 8.57a 2.18 1.11b
F-Value 0.12 0.19 0.51 0.51 0.01 0.7 0.02 0.21 0.03 0.04 0.03 0.01 2.90 0.37 0.01 0.06
P-Value 0.999 0.994 0.855 0.857 1.000 0.704 1.000 0.992 1.000 1.000 1.000 1.000 0.013 0.941 1.000 1.000
* EC: soil salinity (ds m-1, paste extract), S.O.C (g g-1): soil organic carbon content, Ava. N, P, and K (mg g-1): available N, P and K content, SWC (%): soil water content, TBC (log CFU g-1): total bacteria count and TFC (log CFU g-1): total fungi count. UNM: un-mulched, WPM: white plastic mulching, RSM: rice straw mulching, SDM: sawdust mulching. Ch0: distilled water (control), Ch1: 250 mg chitosan L-1, Ch2: 125 mg nano-chitosan L-1 and Ch3: 62.5 mg nano-chitosan L-1.

3.2.4. Soil Nutrients

The soil’s available nitrogen and phosphorus under RSM treatment were significantly higher than other mulch and no-mulch treatments (Table 1). While the highest available potassium contents were obtained under SDM treatment. No significant difference in the soil’s available nitrogen and phosphorus content between the RSM and SDM treatments. There was also no significant difference in the soil available nitrogen content between SDM and WPM treatments.
The application of all chitosan foliar applications had insignificant affect soil available phosphorus and potassium content values in both seasons (Table 1). While the result of soil nitrogen content shows a slight significant change in the 2023 season and don’t changes was found in the 2022 season with the chitosan foliar applications.
Mulching × chitosan foliar applications interaction was not significant soil nitrogen content in both seasons (Table 1). Also, the results showed that the highest soil phosphorus content was associated with organic mulching (RSM and SDM) of the soil with various foliar chitosan sprays (Ch0, Ch1, Ch2, and Ch3). RSM + Ch3 treatment gave the highest values of soil potassium content in both seasons (Table 1), with insignificant differences with other chitosan foliar applications (Ch0, Ch1, and Ch2).

3.2.5. Soil microbial communities

The obtained results in Table 1 showed a significant difference in bacterial and fungal communities in soils under different mulching treatments. Therein, the relative abundance of total bacteria count under SDM treatment (7.98 and 8.33 log CFU g-1) was significantly higher than that under UNM treatment (5.92 and 5.96 log CFU g-1). Also, SDM treatment caused a significantly reduction on the total fungi count than that other mulching treatment.
The chitosan foliar applications presented significant differences in bacterial and fungal communities (Table 1). Foliar application by nano chitosan caused the relative abundances of TBC than that in Ch0 and Ch1, while TFC was significantly decreased in second season due to 3 foliar chitosan applications than that Ch0.
The interactive effect of different mulching materials and chitosan foliar applications was found significant on soil microbial communities (Table 1). The lowest TFC (2.18 and1.11 log CFU g-1) were obtained with SDM + Ch3 treatment. In addition, combine SDM with Ch3 gave the highest TBC in soil.

3.2.6. Principal Component Analysis (PCA)

The PCA biplot illustrated in Figure 2 shows all studied soil parameters on the first two principal components (Dim1= 62.3% and Dim2 =15.3%). It is clear that all studied soil parameters positively correlated with Dim1, except soil EC and total fungi count which showed a negative correlation. Meanwhile, Dim2 exhibited positive correlation with soil water content, soil organic carbon, available N, P, K and total bacteria count and negative correlation with soil EC and total fungi count. Dim1 and Dim2 successfully separated the interactive effect of different mulch materials and foliar applications by chitosan, as seemed to group together. Meanwhile the effect of the control treatments (UNM) varied strongly.

3.3. Common bean yield and its components

3.3.1. Plant Height

Data in Table 2 indicate that the plant height was significantly increased with different mulching materials in comparison with the control. Relative to the control (UNM), the plant height was decreased by 13.44, 16.46 and 13.60% in plants mulched by WPM, RSM, and SDM, respectively. Meanwhile, foliar chitosan application markedly improved plant height and alleviated the adverse salinity effects. In control plants, the plant height reduction was 18.99, 20.07 and 20.98%, compared with the Ch1, Ch2, and Ch3, respectively.
The interactive effects of different mulching materials and foliar applications by chitosan were found significant (at p ≤ 0.05) on the plant height of common beans (Table 2).
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Figure 2. PCA biplot for all studied soil parameters had response to interactive effect of different mulch materials and chitosan foliar applications.
Figure 2. PCA biplot for all studied soil parameters had response to interactive effect of different mulch materials and chitosan foliar applications.
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3.3.2. Dry Weights of Shoots and Roots

All mulching treatments gradually increased the dry weights of both shoot and roots (Table 2), and the minimum values in this respect were recorded with un-mulched treatment (UNM).
In addition, plants sprayed with chitosan had significantly higher dry weights of shoots and roots than control plants. Compared with the control plants, the dry weight of shoot was increased by 16.46 and 14.24% with 62.5 mg nano-chitosan L-1 treated plants (Ch3).
Dry weight of shoot was increased by 31.96 and 26.54% under rice straw mulch treatment plus 62.5 mg nano-chitosan L-1 treated plants (Table 2).
The same direction was noticed in roots, as dry weight was increased by 23.07 and 22.25%, while they were 39.26 and 39.24% in rice straw mulch treatment plus 63 mg nano-chitosan L-1 treated plants, respectively (Table 2).

3.3.3. Seed yield

The different mulch treatments had a statistically similar seed yield in both seasons (Table 2). The highest seed yield (2030.93 and 2075.77 kg ha-1) was obtained with a foliar application of 62.5 mg nano-chitosan L-1 (Ch3). The results evidenced that the interactive effect of different mulch materials plus by chitosan foliar applications gave a statistically similar seed yield in both seasons (Table 2).
Table 2. Plant height, dry weight of shoots and roots, seed yield and nutrients concentration of common bean seed affect under different treatments affect.
Table 2. Plant height, dry weight of shoots and roots, seed yield and nutrients concentration of common bean seed affect under different treatments affect.
Plant parameters Ph (cm) SDW (g) RDW (g) SY (kg ha-1) N (%) P (%) K (%) Na (%)
Seasons 1st 2nd 1st 2nd 1st 2nd 1st 2nd 1st 2nd 1st 2nd 1st 2nd 1st 2nd
Mulching materials
UNM 24.81b 24.91c 2.04b 2.10b 0.206b 0.213b 1450.44b 1481.24b 1.20d 1.24c 0.222d 0.236c 0.659c 0.668d 0.131a 0.119a
WPM 28.14a 28.42ab 2.21a 2.26a 0.232a 0.238a 1981.35a 2014.03a 1.38c 1.45b 0.240c 0.251b 0.985b 1.00c 0.096c 0.077c
RSM 28.89a 29.37a 2.23a 2.29a 0.242a 0.248a 2003.96a 2040.54a 1.47a 1.50a 0.264a 0.272a 1.04a 1.06b 0.102b 0.082bc
SDM 28.18a 28.70b 2.21a 2.28a 0.236a 0.243a 1991.04a 2025.55a 1.43b 1.48ab 0.253b 0.258b 1.06a 1.10a 0.104b 0.085b
F-Value 59.56 80.23 19.73 12.83 36.53 28.07 153.35 154.76 123.47 105.93 61.93 55.47 309.09 389.84 7.92 12.91
P-Value <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001
Chitosan foliar application
Ch0 23.92d 24.22d 1.97c 2.07c 0.199c 0.206b 1543.36c 1565.80c 1.22d 1.29c 0.185d 0.193d 0.770d 0.783d 0.153a 0.137a
Ch1 28.46c 28.75c 2.17b 2.23b 0.233b 0.240a 1918.59b 1942.63b 1.39c 1.43b 0.243c 0.257c 0.913c 0.940c 0.105b 0.090b
Ch2 28.72b 29.09b 2.24ab 2.28ab 0.239ab 0.245a 1933.92b 1977.14b 1.42b 1.46ab 0.269b 0.275b 1.00b 1.03b 0.092b 0.071b
Ch3 28.93a 29.33a 2.30a 2.36a 0.245a 0.251a 2030.93a 2075.77a 1.45a 1.49a 0.280a 0.292a 1.06a 1.08a 0.083b 0.065b
F-Value 102.38 118.73 48.00 25.79 61.86 46.63 96.29 103.85 91.83 61.14 351.76 472.01 140.76 169.01 31.38 37.88
P-Value <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001
Interaction
UNM Ch0 21.70c 21.87d 1.60h 1.78c 0.159f 0.164g 1135.43c 1167.80c 0.952i 0.964i 0.166h 0.168h 0.443k 0.444n 0.204a 0.195a
UNM Ch1 25.33b 25.40bc 2.12fg 2.17ab 0.216de 0.224def 1513.68b 1518.44b 1.20h 1.29h 0.231f 0.250f 0.680j 0.703m 0.115d 0.107c
UNM Ch2 25.86b 25.93bc 2.17ef 2.19ab 0.222cde 0.229cdef 1534.62b 1559.07b 1.30g 1.33gh 0.243de 0.258def 0.742i 0.752l 0.109de 0.093d
UNM Ch3 26.33b 26.43b 2.25bcd 2.27ab 0.227cde 0.232bcdef 1618.09b 1679.64b 1.34fg 1.39efg 0.249de 0.269d 0.772hi 0.775l 0.096gh 0.082e
WPM Ch0 24.43b 24.47c 2.07g 2.14b 0.210e 0.216f 1674.26b 1692.35b 1.23h 1.37fg 0.176h 0.194g 0.806h 0.824k 0.125c 0.108c
WPM Ch1 29.23a 29.63a 2.17ef 2.23ab 0.234bcd 0.241abcd 2040.14a 2069.96a 1.41de 1.45cde 0.240ef 0.252f 0.955ef 0.979hi 0.098fg 0.082e
WPM Ch2 29.40a 29.67a 2.25bcd 2.28ab 0.241abc 0.245abcd 2056.63a 2108.37a 1.43cde 1.47bcd 0.267c 0.270d 1.06cd 1.08ef 0.084ij 0.061f
WPM Ch3 29.50a 29.90a 2.31ab 2.38ab 0.244abc 0.250abc 2154.38a 2185.48a 1.46bcd 1.52abc 0.277b 0.287bc 1.11b 1.13cd 0.076j 0.057f
RSM Ch0 25.07b 25.60bc 2.13fg 2.19ab 0.216de 0.222def 1685.68b 1694.87b 1.37ef 1.43def 0.203g 0.207g 0.911g 0.917j 0.140b 0.120b
RSM Ch1 30.00a 30.27a 2.21cde 2.25ab 0.243abc 0.251abc 2070.60a 2101.06a 1.47bc 1.50abc 0.257cd 0.268de 0.980e 1.02gh 0.100efg 0.086de
RSM Ch2 30.17a 30.70a 2.26abc 2.32ab 0.251ab 0.256ab 2072.20a 2131.53a 1.50ab 1.54ab 0.289b 0.297b 1.10bc 1.11de 0.087hi 0.063f
RSM Ch3 30.33a 30.90a 2.33a 2.40a 0.257a 0.265a 2187.39a 2234.65a 1.53a 1.55a 0.306a 0.316a 1.16a 1.18ab 0.080ij 0.059f
SDM Ch0 24.47b 24.93bc 2.10g 2.16ab 0.212de 0.220ef 1678.07b 1708.20b 1.32fg 1.39efg 0.196g 0.202g 0.918fg 0.945ij 0.142b 0.124b
SDM Ch1 29.27a 29.70a 2.19def 2.26ab 0.239abc 0.244abcd 2049.97a 2081.07a 1.46bcd 1.49abcd 0.246e 0.257ef 1.04d 1.06fg 0.106def 0.088de
SDM Ch2 29.43a 30.07a 2.25bcd 2.31ab 0.243abc 0.250abc 2072.20a 2109.63a 1.46bc 1.50abc 0.278b 0.274cd 1.11b 1.17bc 0.088hi 0.066f
SDM Ch3 29.57a 30.10a 2.31ab 2.39a 0.251ab 0.259a 2163.90a 2203.24a 1.49ab 1.52ab 0.291b 0.298b 1.20a 1.23a 0.082ij 0.061f
F-Value 0.3 0.47 6.79 2.01 2.24 1.96 0.02 0.09 5.23 8.37 1.95 2.76 2.23 2.35 1.41 1.61
P-Value 0.969 0.883 <.0001 0.071 0.046 0.078 1.00 1.00 <.0001 <.0001 0.08 0.017 0.046 0.037 0.221 0.155
* Ph: plant height, SDW: shoots dry weight, RDW: roots dry weight, SY: seed yield. UNM: un-mulched, WPM: white plastic mulching, RSM: rice straw mulching, SDM: sawdust mulching. Ch0: distilled water (control), Ch1: 250 mg chitosan L-1, Ch2: 125 mg nano-chitosan L-1 and Ch3: 62.5 mg nano-chitosan L-1.

3.3.4. Macro-nutrients concentration of common bean seed

The mulch materials had significant (P ≤ 0.05) content of N, p, k, and Na in seeds (Table 2). Rice straw mulch (RSM) increased N and P% in seed, while sawdust mulch (SDM) increased significantly K% and white plastic mulch (WPM) reduced significantly Na% in seed, compared to un-mulched (UNM).
Compared with the control plants, chitosan foliar applications reduced significant Na% in seed and gave the highest NPK concentration in parallel with foliar nano-chitosan at a rate of 62.5 mg L-1 (Ch3). Under the interactive effect of mulch and chitosan foliar applications, the soil covered by both organic mulching materials plus 62.5 mg nano-chitosan L-1 increased N, P, and K, in seed, whereas seed Na decreased significantly with all treatments compared to control plants (Table 2).

3.3.5. Principal Component Analysis (PCA)

Figure 3, loading PCA biplot of all studied plant parameters showed that, the first and two principal components described (Dim1= 90% and Dim2 =3.9 %) of the total variability which noted that Na% in seed had a negative correlation with other studied plant parameters especially shoot and root dry weight. Dim1 and Dim2 successfully separated the interactive effect of different mulch materials and foliar by chitosan applications. This again indicates that chitosan foliar applications s have the largest impact on studied common beans plant parameters, than different mulches treatments. Interestingly the correlation analyses did show a high relationship between all the soil nutrient content values.
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Figure 3. PCA biplot for all studied plant parameters had response to interactive effect of different mulch materials and foliar by chitosan applications.
Figure 3. PCA biplot for all studied plant parameters had response to interactive effect of different mulch materials and foliar by chitosan applications.
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3.4. Correlation matrix

To explore the impact of soil salinity factors on all studied soil and plant parameters, we analyzed the relationships between all studied parameters using a correlation matrix in R (Figure 4). The soil EC was positively related to the abundance of total fungi count and also increased Na content in common bean seeds. The other soil parameters was positively correlated and also with plant parameters. The results showed that the improvement soil characteristics had great influences on common beans productivity.
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Figure 4. Correlation matrix among all studied parameters affected mulch materials and foliar chitosan application.
Figure 4. Correlation matrix among all studied parameters affected mulch materials and foliar chitosan application.
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4. Discussion

4.1. Soil parameters

Salinization causes significant restrictions and decreases crop production due to limitations on various crops [42]. In the current study, mulching and foliar chitosan spraying application a suitable method was used to reduce the effect of salinity on common bean productivity, as well as comparing different materials of mulching on some chemical and biological soil characteristics. The results showed that the use of white plastic mulch (WPM) led to a decrease in soil salinity and in addition to an increase in the water content in the soil. From this result, the decrease in soil salinity could be linked to an increase in moisture content. These results are consistent with [18,43,44,45], they confirmed that the mulching procedures not only boosted soil water content, but also contributed to the leaching of soil salt and reducing the soil salt content; this is attributed to the decrease in soil evaporation.
The increase in the organic carbon content of the soil under organic mulching could be attributed to the increase in organic matter content in the soil due a higher decomposition rate, thereby increasing the organic carbon content [46]. The present finding conformed to Hossen et al., [47]; they indicated that the sawdust mulching treatment increased soil organic carbon more than other mulching treatments studied.
The increase in available nutrients in soils is a result of increased microbial activity and soil moisture content beneath the mulching materials which led to an improvement in soil fertility. Xiaomin [48] indicated that the positive changes in the available nutrients due to organic mulching can be attributed to the increased biological activities, thus, giving rise to the mineralization of organic matter.
The results showed that, the application of different materials of mulching had a contradictory effect on the bacterial communities in the soil, as the numbers of bacteria increased due to the presence of organic carbon, and in contrast, the numbers of fungi in the soil decreased as a result of mulching treatments. These results are consistent with previous studies for both [20,49].
The results revealed that the foliar application of chitosan had a positive effect on soil salinity and carbon content. This can be attributed to helping in the spread of roots through the soil layers, as it led to loosening of the soil, which helped to get rid of salts. Also, root residues in the soil led to an increase in organic matter, which was reflected in an increase the soil organic carbon, and consequently the bacterial populations. However, the total fungi count in the soil was significantly decreased due to chitosan foliar applications. The present finding was conformity with [50,51]. Chitosan or its nanoparticles foliar application have been reported to harm fungi growth by induced defense responses in plants against defense system against the pathogen [52,53]. In addition, these negative effects could be due to the repeated amino groups of the chitosan structure [54], the external electrostatic interaction between the positive amino glucosamine groups –NH3+ of chitosan and phospholipids in the fungal cell membrane, leads to changes in cell permeability and leakage of intracellular electrolytes and proteinaceous constituents and cell death [55].

4.2. Common bean yield and its components

The salt stress had a noxious effect on the growth, physiological, and productivity of common bean [56]. According to Assimakopoulou et al., [57], the common bean (Phaseolus vulgaris L.) is considered a salt-sensitive plant, and the 0 - 75 mM NaCl concentration caused biomass and yield reduction. The use of mulching in these conditions led to improved growth and this was reflected in the crop as a result of improved soil characteristics, including a decrease in Salinity, soil fertility, and moisture availability. The results obtained are consistent with [21,22,23].
The improvement in growth parameters and common yield can be attributed to the use of chitosan which is more active against salt stress by reducing oxygen free radicals or blocking ROS activity, promoting cell division, increasing ionic transport, polyamine content, and membrane stabilization under stress conditions [58]. The stimulating effect of chitosan on plant growth may be attributed to an increase in the availability and uptake of water and essential nutrients via cell osmotic pressure adjustment, as well as a reduction in the accumulation of harmful free radicals (ORS) via increased antioxidants and enzyme activities [59]. Zayed et al., [61] study the effect of nano-chitosan application on Phaseolus vulgaris under salinity stress, and found that the plant height and dry weights of the shoots increased significantly with nano chitosan application as a result in increasing of antioxidant enzymes. Also, the application of CSNPs improves chlorophyll content and plant metabolism in salt-stressed mung beans (Phaseolus vulgaris L.), as evidenced by a reduction in malondialdehyde and H2O2 contents [60].

5. Conclusion

It can conclude that, the beneficial effects of mulching materials have effective for inhibiting salt accumulation and increasing soil water and nutrient content. The foliar spray applications of 62.5 mg nano chitosan L-1 at 15, 30, and 45 DAS alleviated the salt stress and improved the growth of common bean plants. For sustainable eco-system, combining mulching with foliar chitosan spray application can be an effective practice for the inhibition of salt accumulation and improved growth and productivity of common bean grown in salt-affected soil.

Author Contributions

Conceptualization, N.A., M.A., T.K., and E.S.; methodology, N.A., M.A., T.K., and E.S.; software, T.K.; validation, N.A., M.A., T.K., and E.S.; formal analysis, T.K., E.S. and A.E-D.O.; investigation, N.A., M.A., T.K., and E.S.; resources, , N.A., M.A., T.K., E.S., M.E., and A.E-D.O.; data curation, N.A., M.A., T.K., and E.S.; writing—original draft preparation, , T.K., and E.S.; writing—review and editing, N.A., M.A., and A.E-D.O.; visualization, N.A., M.A., and E.S.; supervision, T.K.; funding acquisition, M.E. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data sets analyzed during the present study are accessible from the current author on reasonable request.

Acknowledgments

The authors extend their appreciation to the Deanship of Scientific Research at King Khalid University for funding this work through the Larg Groups Project under grant RGP2/164/44. All the authors extend their esteem, Soil, Water, and Environment Res. Inst., Agriculture Research Center, Giza, Egypt, Soil and Water Department, and Horticulture Department Faculty of Agriculture, Tanta University, Tanta, Egypt. Also, thankful for the support provided by Labs of soil improvement and conservation Res. Dep., and soil microbiology Res. Dep., Sakha Agri. Res. station, Kafr El-Sheikh, Egypt.

Conflicts of Interest

The authors declare no conflict of interest.

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Preprints 94181 g001
Figure 1. Daily rainfall and temperature during the experimental period of common bean crop (2022 and 2023).
Figure 1. Daily rainfall and temperature during the experimental period of common bean crop (2022 and 2023).
Preprints 94181 g001
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