Next Article in Journal
Differentially Colored Photoselective Nets as a Sophisticated Approach to Improve the Agronomic and Fruit Quality Traits of Potted Blueberries
Previous Article in Journal
Assessment of Interstitial Woody Plantings in North–South Axis Solar-Tracking Photovoltaic Systems: Balancing Shading Impact and Landscape Integration
Previous Article in Special Issue
Atractylodes macrocephala Root Rot Affects Microbial Communities in Various Root-Associated Niches
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Agronomy
Volume 15
Issue 3
10.3390/agronomy15030695
agronomy-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

A Bacterial Endophyte Bacillus amyloliquefaciens W10 Enhances the Tomato Resistance Against Tuta absoluta

by
Mingshi Qian
†,
Chaoqi Sheng
†,
Mingying Zheng
,
Ke Zhu
,
Youxin Yu
,
Gang Xu
* and
Guoqing Yang
*
College of Plant Protection, Yangzhou University, Yangzhou 225009, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Agronomy 2025, 15(3), 695; https://doi.org/10.3390/agronomy15030695
Submission received: 5 February 2025 / Revised: 27 February 2025 / Accepted: 12 March 2025 / Published: 13 March 2025
(This article belongs to the Special Issue Molecular Advances in Crop Protection and Agrobiotechnology)
Browse Figures
Versions Notes

Abstract

:
The tomato leafminer, Tuta absoluta, is a destructive invasive tomato pest worldwide. Bacillus amyloliquefaciens W10, a bacterium isolated from the rhizosphere of tomatoes, is classified as a plant growth-promoting rhizobacterium. However, whether B. amyloliquefaciens W10 can improve the resistance of tomato against T. absoluta remains unclear. In this study, we found that B. amyloliquefaciens W10 promoted the tomato growth and significantly reduced the fecundity of T. absoluta. To further evaluate the effects of B. amyloliquefaciens W10 on the tomato’s resistance to T. absoluta, the age-stage, two-sex life table, and oviposition preference test were carried out to investigate the individual fitness, population parameters, and preference behavior of T. absoluta. Compared to the control, the intrinsic rate of increase (rm), net reproductive rate (R0), and finite rate of increase (λ) of T. absoluta in the treatment group were significantly reduced, while the population doubling time (DT) was significantly increased. Meanwhile, the oviposition preferences of T. absoluta for the treated tomato plants were reduced. After T. absoluta infestation, electrical conductivity and hydrogen peroxide (H2O2) content in tomato leaves after B. amyloliquefaciens W10 treatment were significantly lower than those in the control, while peroxidase (POD), polyphenol oxidase (PPO), jasmonic acid (JA), and salicylic acid (SA) levels were significantly higher. In addition, the O2−, superoxide dismutase (SOD), and catalase (CAT) levels were also affected. qRT-RCR analyses revealed that B. amyloliquefaciens W10 colonization induced the expressions of JA- and SA-related genes, including AOS1, AOC, PAL1, and SAMT, in tomato plants after T. absoluta infestation. These findings provide valuable insights and theoretical support for the application of beneficial endophytes to induce the resistance in tomatoes against T. absoluta.

1. Introduction

The tomato leafminer Tuta absoluta is a globally invasive pest with destructive damages to tomato crops, originating from Peru in South America, invading Spain in Europe in 2006, and then spreading rapidly to some countries in Europe, Africa, South America, Asia, and other places [1,2,3,4]. T. absoluta begins infesting plants by laying eggs on the upper plant organs. Larvae feed on the leaf mesophyll, damaging the leaves and reducing the photosynthetic capacity and yield of the plant. In the absence of effective means of control, the consequences of pest infestation can lead to serious yield reductions, with the losses of 20% to 30%, heavy more than 50%, or even crop failure [5,6]. In response to the invasion and damage of T. absoluta, fast-acting and effective chemical control remains the mainstay in most countries and regions, ease of use and rapid control of pest populations, but it also poses a number of problems. Because T. absoluta feeds on the leaves and can also hide in young stems and branch tips, it leads to additional spraying and application of insecticides, which greatly reduces efficacy levels [7]. With the continuous spread of T. absoluta and the continuous development of pesticides, there was a wider choice of insecticides against T. absoluta, such as polymyxins, bisamides, benzoylureas, etc., [8,9]. However, the problem of resistance has inevitably arisen due to heavy use, which develops in different geographic populations of pests [10,11]. Therefore, it is urgent to develop eco-friendly and sustainable approaches to control these invasive insect pests.
At present, beneficial microorganisms are widely used as the main biological control agents in plant protection [12,13,14,15]. As an important research direction in microbiological control, beneficial endophytes have great potential as a biocontrol agent, as they can improve plant uptake of soil nutrients, promote plant development, enhance plant stress tolerance, and protect plants from pathogens and insects [16,17,18,19]. Bacterial endophytes have a wide range of host species and play a great role in plant protection without causing harm to plants, and many studies have demonstrated that they can improve the crop’s resistance against diseases and insect pests through endophytic colonization [20,21,22,23]. Among the endophytic bacteria, Bacillus, Azotobacter, Serratia, etc., mainly colonize the inter-root or root cortex surface of the plants, while Rhizobium, Bradyrhizobium, Allorhizobium, etc., are mainly found in specific parts of root cells, and they can not only produce growth-promoting effects on crops, but also induce systemic resistance in plants, which protects the plants from phytopathogenic microorganisms and phytophagous pests [24,25,26,27].
Bacillus amyloliquefaciens is a potential plant growth-promoting rhizobacteria known to inhabit many plants and imparts beneficial effects by enhancing plant growth and contributing to disease resistance, and it can rob nutrients and growth space from pathogenic bacteria, metabolize products that inhibit pathogen reproduction, promote plant growth, and induce plant resistance [14,28,29,30,31]. A variety of B. amyloliquefaciens strains have been isolated from various microbiomes, including the rhizosphere, endophytic environments, the phyllosphere, and soil microbiomes [32,33,34]. In the area of plant growth promotion, B. amyloliquefaciens DB2 significantly promoted the growth of wheat seedlings, including the fresh weight, dry weight, height, and root length [35]. Khan et al. found that five kinds of B. amyloliquefaciens isolates enhanced Nicotiana benthamiana growth parameters, including shoot and root length, leaf number, area, and biomass [36]. Meanwhile, the use of B. amyloliquefaciens for disease resistance also has been investigated in plants. In Lycium barbarum, B. amyloliquefaciens HSB1 and FZB42 had remarkable inhibition activity against Fusarium oxysporum that frequently causes root rot [37]. In Arabidopsis thaliana, B. amyloliquefaciens strain GD4a isolated from switchgrass produced a functional bacterial extracellular exudate that disrupted the pathogenicity of Botrytis cinerea [34]. B. amyloliquefaciens could also act as a potential biocontrol agent to control insect pests. The use of B. amyloliquefaciens YJNbs21.10 had a good inhibitory efficacy against Rhopalosiphum padi [38]. B. amyloliquefaciens MZ895491 could affect the growth and development of the corn pest Spodoptera frugiperda and reduce its feeding ability and improve the metabolic functions of corn leaves [39]. Additionally, B. amyloliquefaciens-mediated regulations related to biotic stresses have been explored in plants. Qian et al. reported that the colonization of B. amyloliquefaciens AK-12 upregulated the expressions of resistance genes related to peroxidase (POD), phenylalanine ammonia-lyase (PAL), and polyphenol oxidase (PPO) in Brassica napus [40]. Qi et al. suggested that B. amyloliquefaciens KRS005 colonization could upregulate the defense-related genes of reactive oxygen species (ROS), jasmonic acid (JA) and salicylic acid (SA)-related signaling pathways to enhance the resistance to gray mold of tobacco [41]. Nevertheless, there are still some limitations in understanding the effects of B. amyloliquefaciens on plant resistance against insect pests.
B. amyloliquefaciens W10 was previously isolated from the rhizosphere of tomato [42]. In this study, we investigated whether B. amyloliquefaciens W10 could promote the growth of tomato and enhance the resistance to T. absoluta. First, we determined the growth indexes of tomato, as well as the survival rate and fecundity of T. absoluta after root irrigation with B. amyloliquefaciens W10. Then, fitness-related traits and population parameters of T. absoluta on tomato treated by B. amyloliquefaciens W10 were estimated using the age-stage, two-sex life table. In addition, the oviposition preference of T. absoluta was used to further assess the effects of B. amyloliquefaciens W10. Finally, the physiological and biochemical indexes of tomato were measured after the treatment with B. amyloliquefaciens W10, and the expressions of JA- and SA-related genes were analyzed to explore the underlying mechanism of B. amyloliquefaciens W10-mediated tomato defense against T. absoluta. This study will facilitate the development of beneficial endophytes in integrated pest control and provide new technical and theoretical support for the application of green control systems in crop protection.

2. Materials and Methods

2.1. Plant Material and Insects

The tomato variety Zhongshu No. 4 was cultured in an artificial climate chamber with parameters set as follows: 27 ± 1 °C, 60 ± 5% relative humidity, a photoperiod of 16 h:8 h (L:D). The T. absoluta strain was provided by Institute of Plant Protection, Chinese Academy of Agricultural Sciences, and has been continuously reared for many generations. The following experiments were conducted from September 2022–May 2024.

2.2. The Cultivation and Inoculation of B. amyloliquefaciens W10

B. amyloliquefaciens W10 isolated from the rhizosphere of tomato was provided by Prof. Qingxia Zhang (Yangzhou University) [42]. B. amyloliquefaciens W10 was cultured in tryptic soy agar (TSA) (Fangxin, Guangdong, China) medium at 28 °C for 24 h and recovered on TSA at 50 °C after 12 h. The bacterial solution was diluted for testing.
Tomato seeds were sterilized by soaking in 75% ethanol solution for 15 s, washed three times with ddH2O, followed by soaking in 10% NaClO solution for 15 s, and the rinsing was repeated for five times. Finally, tomato seeds were soaked in ddH2O for 15 h and sprouted in a dark environment at 28 °C for 36 h. Seedlings were raised in 5 × 10 seedling trays, and tomato seedlings of the same growth were selected and transplanted into round, transparent plastic cups 5 cm in diameter and 10 cm in height containing the nutrient soil, one tomato seedling per cup. The experiment was divided into seven groups, including one control group and six treatment groups. Thirty plants per group were used for the experiment. After 20 days of tomato growth, six treatment groups were irrigated with 10 mL bacterial solution B. amyloliquefaciens W10 (108 CFU/mL, 2 × 108 CFU/mL, and 4 × 108 CFU/mL), respectively, and the control group was treated with water. After 5 days, three treatment groups were selected for the second root irrigation.
To evaluate the colonization of B. amyloliquefaciens W10 in the tomato roots, the total RNA was isolated from the roots of tomato plants using the TRIzol reagent (Invitrogen, Carlsbad, CA, USA). cDNA was synthesized with the HiScript III 1st Strand cDNA Synthesis Kit (Vazyme, Nanjing, China) [43]. The primers for B. amyloliquefaciens W10 (Table S1) were designed, and RT-PCR was used to determine the colonization of B. amyloliquefaciens W10 in the roots.

2.3. Determination of Tomato Growth Parameters

The experiment was divided into seven groups as described in Section 2.2. The tomato plants were harvested 20 days after the first root irrigation, and the growth parameters of tomato seedlings were measured, including stem diameter, stem height, shoot fresh weight and root fresh weight. Stem diameter was measured 5 cm above the lateral roots of the tomato plants. Stem height was measured starting 5 cm above the lateral roots of tomato plants and ending at the level of the uppermost leaves. Shoot fresh weight and root fresh weight were measured using the soil as the dividing point. Thirty plants per group were used to measure.

2.4. Analysis of Biological Parameters in T. absoluta

T. absoluta eggs were attached one egg per cup to the leaves of tomato grown to three leaves stage, each treatment had 100 replicates, and pupation and survival of larvae and pupae were recorded every day. A male and a female adult from the same treat group were mated and placed together in a round clear plastic cup. Cotton balls containing 5% honey water and fresh leaves were replaced daily. Egg production was recorded every day. The TWOSEX-MSChart 2.00.2723 program was applied to calculate the population life table parameters. The variances and standard errors of population parameters were calculated by the bootstrap method with 100,000 re-samplings, and the differences among the different groups were determined by paired bootstrap test (p < 0.05). The population projections of T. absoluta for 60 days were performed with the TIMING-MSChart 2.00.0331 program. The population projection of each treatment was begun with 10 individuals for comparative purposes [44,45,46].

2.5. Oviposition Choice Test

Three cups of untreated tomato plants and three cups of B. amyloliquefaciens W10-treated plants (2 × 108 CFU/mL) were placed together in a mesh cage made of nylon mesh and placed two by two crosswise to form a hexagonal shape. Six pairs of T. absoluta that have emerged one day were placed on Petri dishes to move freely, and the number of eggs on each tomato seedling was counted on the fifth day.

2.6. Determination of Electrical Conductivity and MDA

Electrolyte leakage and MDA content were measured in B. amyloliquefaciens W10-treated tomato leaves at different time intervals (0, 1, 3, 5, and 7 days) after T. absoluta infestation. Electrolyte leakage was determined as previously described [47]. The leaves were collected from 10 distinct plants and cut into 4 cm2 pieces. They were placed in 5 mL ddH2O for 30 min, and the conductivity was determined as (C1) after standing for 1 h at room temperature. Then, it was placed in a 100 °C water bath for 5 min, transferred to the outside and cooled to room temperature, at which time the solution conductivity was determined as (C2). The results were analyzed using the formula (C1/C2) × 100%.
The MDA content was measured via the reaction of thiobarbituric acid (TBA) [48]. The leaves used in each group were collected from 10 distinct plants and homogenized through grinding. Approximately 0.1 g of tomato leaves was homogenized in 1 mL of 10% (w/v) trichloroacetic acid (TCA). The mixture was centrifuged at 10,000× g for 10 min at 4 °C. The supernatant was mixed with an equal volume of TBA (0.6% in 10% TCA) and then boiled at 100 °C for 20 min and rapidly cooled on ice. After centrifuging at 3000× g for 10 min at 4 °C, the supernatant absorbance was determined at 532 nm and corrected for non-specific absorption at 600 nm.

2.7. Determination of H2O2 and O2− Content

Endogenous concentrations of hydrogen peroxide (H2O2) and oxygen ion (O2−) were measured as previously described with the H2O2 and O2− assay kit (Solarbio, Beijing, China) [49]. Approximately 0.1 g of tomato leaves inoculated with B. amyloliquefaciens W10 after T. absoluta infestation was mixed with 1 mL of reagent 1. The mixture was homogenized in an ice bath and centrifuged at 8000× g for 10 min at 4 °C. Then, the supernatant was tested to establish the calibration curves. 250 μL of sample, 25 μL of reagent 2 and 50 μL of reagent 3 were placed into an EP tube, and then centrifuged at 4000× g at room temperature for 10 min. The supernatant was removed, and 250 μL of reagent 4 was added. Finally, 200 μL of dissolved sample was transferred to a 96-well plate, and the absorbance was determined using a microplate reader.

2.8. Determination of Defense-Related Enzymes Activities

The activities of POD, psuperoxide dismutase (SOD), catalase (CAT), glutathione S-transferases (GST), PAL, and PPO were determined in the B. amyloliquefaciens W10-treated tomato leaves at different time intervals (0, 1, 3, 5, and 7 days) after T. absoluta infestation. The activities of these defense-related enzymes were determined using the assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer’s instructions.

2.9. Determination of JA and SA Content

The second instar larvae of T. absoluta were starved for 2 h and fed on tomatoes grown for 30 days from the control and B. amyloliquefaciens W10 treatment groups, and the tomato leaves were collected after 12 and 24 h. The tomato leaves were then ground into powder with liquid nitrogen, and 50 mg of the sample was placed in a 2 mL centrifuge tube, to which 500 μL of extractant (Isopropanol:ddH2O:HCl = 2:1:0.002) was added. The tube was shaken at 100 r/min and 4 °C for 30 min. 1 mL of dichloromethane was added. The tube was shaken for 30 min at 4 °C. Then, the suspension was centrifuged at 12,000× g at 4 °C for 5 min. 900 μL of the lower phase was freeze-dried, and 100 μL of methanol was added to dissolve the sample. Detection of JA and SA was performed using the assay kits (Shanghai Qiaoshe Biotechnology Co., Shanghai, China) according to the manufacturer’s instructions.

2.10. Gene Expression Analysis

Tomato leaves were collected for RNA extraction to perform qRT-PCR. The total RNA was isolated using the TRIzol reagent (Invitrogen, Carlsbad, CA, USA). cDNA was synthesized with the HiScript III 1st Strand cDNA Synthesis Kit (Vazyme, Nanjing, China). qRT-PCR was performed on a CFX 96 Real-Time Detection System (Bio-Rad, Hercules, CA, USA) with the ChamQ SYBR qPCR Kit (Vazyme, Nanjing, China) [50]. The reference gene ACTIN7 was applied to normalize the expressions of the target genes, and the relative quantifications were calculated using the 2−ΔΔCT method [51]. The qRT-PCR primers are listed in Table S1.

2.11. Statistical Analysis

One-way analysis of variance (ANOVA) followed by Tukey’s honestly significant difference (HSD) test was performed to analyze the biological parameters (p < 0.05). Student’s t-test was used to determine the statistical differences between two groups (* p < 0.05, ** p < 0.01, *** p < 0.001). All data were statistically analyzed with SPSS 16.0 (SPSS Inc., Chicago, IL, USA).

3. Results

3.1. Effects of B. amyloliquefaciens W10 on the Growth of Tomato Seedlings, the Survival Rate and Fecundity of T. absoluta

After root irrigation with B. amyloliquefaciens W10, the colonization of B. amyloliquefaciens W10 in the tomato roots could be detected by RT-PCR (Figure S1). The growth parameters of tomato seedlings after treatment with different concentrations of B. amyloliquefaciens W10 (108 CFU/mL, 2 × 108 CFU/mL and 4 × 108 CFU/mL) were determined (Table 1). B. amyloliquefaciens W10 treatment had no significant effect on the stem diameter of tomato seedlings. 2 × 108 CFU/mL and 4 × 108 CFU/mL root irrigation two treatment groups had significantly higher stem height than the control group. Consistent with the trend of stem height, the shoot fresh weight was significantly higher after twice treatments of 2 × 108 CFU/mL and 4 × 108 CFU/mL. The root fresh weight was significantly increased after root irrigation once treatment of 4 × 108 CFU/mL, and all significantly increased under the twice treatments of three concentrations. These results showed that B. amyloliquefaciens W10 could promote the growth of tomato seedlings.
No significant differences were observed in the survival rate of T. absoluta at larval, pupal and pre-adult stages after treatment with B. amyloliquefaciens W10, but the fecundity was affected. Notably, the fecundity of T. absoluta was significantly reduced by twice treatments of 2 × 108 CFU/mL, with a reduction of 35.8% (Table 2).

3.2. Effects of B. amyloliquefaciens W10 on the Biological Parameters of T. absoluta

To further investigate the effect of B. amyloliquefaciens W10 on T. absoluta, the biological parameters of T. absoluta fed on the control tomato plants and B. amyloliquefaciens W10-treated plants (2 × 108 CFU/mL twice) were determined (Table S2). Compared with the control, the B. amyloliquefaciens W10-treated plants had no significant effects on the larval stage, pupal stage, preoviposition period, oviposition period, and adult longevity of T. absoluta (Table S2). Meanwhile, the age-stage specific survival rates (sxj) and life expectancy (exj) of T. absoluta suggested no significant differences between B. amyloliquefaciens W10 treatment and the control (Figures S2 and S3). Age-stage reproductive value (vxj) curve showed that the vxj value of T. absoluta was lower on B. amyloliquefaciens W10-treated tomato (Figure 1). The highest vxj value of T. absoluta females was the 19th day on the control tomato (89.138), while the highest vxj value was the 21st day on the treated tomato (55.393), supporting that the tomato plants colonized by B. amyloliquefaciens W10 could reduce the fecundity of T. absoluta.

3.3. Effects of B. amyloliquefaciens W10 on the Population Life Table Parameters of T. absoluta

The net reproductive rate (R0) of T. absoluta reared on B. amyloliquefaciens W10-treated tomato (2 × 108 CFU/mL twice) was significantly decreased, compared with the control group (Table 3). T. absoluta feeding on B. amyloliquefaciens W10-treated tomato exhibited a significant reduction in the intrinsic rate of increase (rm) and finite capacity of increase (λ), but an increase in doubling time (DT). No significant difference in the mean generation time (T) was observed under the treatment of B. amyloliquefaciens W10 (Table 3).
Projected population growth for T. absoluta feeding on B. amyloliquefaciens W10-treated tomato plants was shown in Figure 2. Estimated population growth of T. absoluta after B. amyloliquefaciens W10 treatment (4.02) were lower than the control (4.37). The starting population of T. absoluta was set at 10 eggs. There were 7702 eggs, 573 females and 296 males in the control group; while there were 2793 eggs, 171 females and 197 males in the treatment group (Figure 2).

3.4. Effects of B. amyloliquefaciens W10 on the Oviposition Preference of T. absoluta

To investigate the effect of B. amyloliquefaciens W10 on the oviposition preference of T. absoluta, we compared the number of eggs laid on the tomato leaves between the control group and the treatment group on the fifth day. The proportion of egg per plant of T. absoluta on the treatment group was significantly lower than the control group (Figure 3). This result suggested that B. amyloliquefaciens W10 colonization in tomato plants deterred ovipositing by T. absoluta.

3.5. Effects of B. amyloliquefaciens W10 on Electrical Conductivity, MDA and ROS Accumulation in the Tomato Leaves

The levels of electrolyte leakage were significantly lower in B. amyloliquefaciens W10-treated tomato leaves than in the control tomato leaves at 5 and 7 days post-treatment (Figure 4A). Meanwhile, no obvious differences in MDA contents were observed between the control and B. amyloliquefaciens W10-treated tomato leaves (Figure 4B).
The production of ROS is usually associated with plant cell death and enhanced susceptibility [52]. H2O2 and O2− are two important ROS. The levels of H2O2 were significantly lower than the control at different time intervals (1, 3, 5, and 7 days) after B. amyloliquefaciens W10 treatment (Figure 5A). However, the levels of O2− were significantly higher than that of the control at 1 day after B. amyloliquefaciens W10 treatment, and lower than that of the control (3 and 5 days) (Figure 5B).

3.6. Effects of B. amyloliquefaciens W10 on the Activities of Defense-Related Enzymes in Tomato Leaves

To investigate whether B. amyloliquefaciens W10 could affect the defense-related enzyme activities of the tomato leaves, the activities of POD, SOD, CAT, GST, PAL, and PPO were measured (Figure 6). The results showed that the activity of POD in the B. amyloliquefaciens W10-treated tomato leaves increased significantly at different time intervals (1, 3, 5, and 7 days) (Figure 6A). The activities of SOD and CAT in the treatment group were significantly higher than those of the control group at 1 and 3 days, while they were lower than those of the control at 5 and 7 days (Figure 6B,C). No significant differences in the activities of GST and PAL were detected between the treatment and control (Figure 6D,E). The PPO activity was significantly increased in the B. amyloliquefaciens W10-treated tomato leaves than the control at 3, 5, and 7 days (Figure 6F). These results indicated that B. amyloliquefaciens W10 colonization stimulated the defense-related enzymes in tomato (Figure 6).

3.7. Effects of B. amyloliquefaciens W10 on JA and SA in Tomato Leaves

The contents of SA and JA were measured in the control and B. amyloliquefaciens W10-treated tomato leaves at different time intervals (0, 12, and 24 h) after T. absoluta infestation. The results indicated that the JA content elevated continuously after T. absoluta infestation and was significantly higher than the control at 12 h and 24 h (Figure 7A). In addition, the SA content of the treatment group increased at 24 h and was significantly higher than that of the control group (Figure 7B).
To further explore the role of B. amyloliquefaciens W10 in the tomato resistance against T. absoluta, we examined the expressions of six genes related to JA (AOS1, AOS2, AOC) and SA (PAL1, SABP2, SAMT) [41,53,54] in tomato leaves after 24 h feeding by T. absoluta (Figure 8). The data demonstrated that B. amyloliquefaciens W10 induced the expression levels of AOS1 (3.81-fold), AOC (0.388-fold), PAL1 (3.37-fold), and SAMT (1.39-fold) compared with the control (Figure 8A,C,D,F). However, no obvious differences in AOS2 and SABP2 expression levels were detected (Figure 8B,E). These results implied that B. amyloliquefaciens W10 colonization activated JA and SA signaling pathways in tomato plants.

4. Discussion

With the massive use of chemical pesticides, the problem of pest resistance is becoming increasingly serious, and the use of rhizobacteria or endophytes to develop plant growth regulators is a current research hotspot [11,17,18]. As T. absoluta damage becomes more severe, traditional means of control become more difficult to limit its invasive expansion. Many studies have demonstrated the ability of endophytes to promote plant growth and increase their resistance [34,37,38,55]. In this study, we analyzed the effects of B. amyloliquefaciens W10 on tomato plant growth and resistance to T. absoluta, providing a potential method for controlling T. absoluta.
To find out the optimal treatment and concentration for controlling T. absoluta and promoting tomato growth, we investigated the effects of different treatments of B. amyloliquefaciens W10 on the tomato growth and T. absoluta. The results showed that B. amyloliquefaciens W10 (2 × 108 CFU/mL, and 4 × 108 CFU/mL) root irrigation twice had the promotive effect on tomato stem height, shoot fresh weight and root fresh weight (Table 1). Similarly, B. amyloliquefaciens DB2 increased the height, root length, fresh weight, and dry weight of wheat seedlings [35]. Additionally, it has been found that the colonization of B. amyloliquefaciens and Bacillus subtilis promoted N. benthamiana growth, including shoot and root length, leaf number, area, and biomass [36]. These results suggest that colonization by endophytic bacteria may interact with the plant to produce certain chemicals to promote plant growth. We first explored the survival and fecundity of T. absoluta fed on the tomato after B. amyloliquefaciens W10 treatment and found that the survival rate was not affected, but the fecundity was significantly reduced (Table 2). These results showed that B. amyloliquefaciens W10 promoted the tomato growth and induced its resistance, leading to a reduction in the fecundity of T. absoluta.
The life table is a fundamental tool for investigating the survival, development, and reproduction of insect populations under abiotic and biotic environmental conditions [45,56]. The age-stage, two-sex life table of T. absoluta after feeding on B. amyloliquefaciens W10-treated tomato plants was established. B. amyloliquefaciens W10 may not be directly harmful to T. absoluta, but rather indirectly induce the resistance. The biological parameters and vxj curve analyses indicated that B. amyloliquefaciens W10 treatment reduced the fecundity of T. absoluta (Figure 1). Similarly, Bacillus brassicae notably reduced the fecundity of Brevicoryne brassicae [57]. These studies suggested that beneficial bacteria colonization could negatively affect the fecundity of insect pests and have potential for pest control.
The population life table parameters rm, R0, T, λ, and DT are applied to evaluate the ability of insect populations to grow under specific environmental conditions, and they are often affected by changes in host plants and environmental conditions [58,59,60]. Our study showed that tomato plants treated with B. amyloliquefaciens W10 significantly decreased the population parameters (rm, R0, and λ) and increased the DT of T. absoluta (Table 3). Our study provided the first report for the negative effects of B. amyloliquefaciens W10 treatment on the population growth of an insect pest. A similar study has shown that five entomopathogenic fungi-colonized tomato plants, all significantly reducing the rm, R0, and λ values of T. absoluta [55]. Simulation of population dynamics using Timing-MSChart 2.00.0331 software also reflected that the population size of T. absoluta fed on B. amyloliquefaciens W10-treated tomato was smaller than that of the control (Figure 2). Similarly, Piriformospora indica-colonized rice plants significantly reduced the population size of Nilaparvata lugens [61]. Taken together, it can be inferred that the colonization of endophytes may trigger plant defense responses, thereby suppressing the population growth of pests.
Insect pests have evolved complex behavioral and sensory mechanisms to select suitable hosts for oviposition, while the interactions between the endophytes and plants may affect the oviposition preference of insect pests [16,62,63,64]. Our study showed that T. absoluta females preferred to oviposit on untreated tomato plants rather than B. amyloliquefaciens W10-treated plants (Figure 3). This may be due to endophyte–plant interactions mediated by the defenses in tomato plants, which may induce volatile organic compound emissions from tomato. In the other choice experiment, S. frugiperda was found to prefer untreated leaves over B. amyloliquefaciens-treated leaves [21]. Similarly, Salazar et al. found that T. absoluta clearly showed a preference for ovipositing on non-inoculated tomato plants compared to those inoculated with Metarhizium robertsii and B. amyloliquefaciens [12]. Therefore, we concluded that endophyte colonization is a potential biocontrol method for plant protection.
Treatment of plants with rhizobacteria leads to structural changes in the cell wall and physiological and biochemical changes, which result in the synthesis of proteins and chemicals involved in the defense mechanisms of the plant [65]. Therefore, we explored the changes in physiological and biochemical indicators related to plant resistance. Electrolyte leakage can reflect the membrane concentration on both sides of plant cells and is the key to determining the membrane permeability, while MDA is considered to be an indicator of cell membranes oxidative damage [66,67,68,69]. In this study, we found that electrolyte leakage decreased in tomato leaves after B. amyloliquefaciens W10 treatment at 5 and 7 days, and the MDA content was not changed (Figure 4A,B). For example, the electrolyte leakage and MDA contents decreased significantly in nectarine fruit with Monilinia fructicola infection after Bacillus licheniformis W10 treatment [70]. Therefore, these results suggested that endophyte colonization had a restorative effect on oxidative membrane damage in plants and decreased oxidative damage.
ROS plays a crucial role in abiotic and biotic stress perception, integration of different environmental signals, and rapid response mechanisms in response to stress, thus contributing to the establishment of defense responses [35,71,72]. H2O2 plays an important role in the plant systemic defense signaling pathways, while O2− mainly regulates plant photosynthesis [73,74]. In this study, we found that after treatment with B. amyloliquefaciens W10, the H2O2 content in the leaves was significantly reduced, indicating that the treatment could effectively remove ROS in tomato leaves (Figure 5A). Other studies have shown that ROS-related signal pathways were significantly upregulated when tobacco leaves were treated with B. amyloliquefaciens-KRS005 [41]. Thus, we inferred that endophyte colonization exerted antioxidant properties, thereby reducing oxidative damage to plant cells and increasing plant resistance.
Various defense-related enzymes play an important role in plant defense responses [75,76]. Our results indicated that the activities of POD and PPO were enhanced by B. amyloliquefaciens W10 treatment (Figure 6). It has been shown that Arbuscular mycorrhizal could enhance the resistance of host plants by inducing the activities of SOD, POD and CAT [77,78]. A similar study has also shown that Bacillus mycoides-inoculated plants enhanced the oxidative defense system by increasing the activities of CAT, glutathione peroxidase (GPX), ascorbate peroxidase (APX), and PAL and could induce secondary metabolites to higher levels [79]. In N. benthamiana, the colonization of B. amyloliquefaciens and B. subtilis also elevated the activities of PPO, POD, and SOD [36]. These results indicated that endophyte colonization could strengthen the enzyme activities of plants to enhance resistance.
The phytohormones JA and SA have been involved in the regulation of signaling pathways related to plant resistance to insect damage as key signals associated with plant stress defense and development in recent decades [34,41,59]. Our results revealed that JA and SA contents in the B. amyloliquefaciens W10 treatment group were significantly higher than those in the control group after T. absoluta infestation (Figure 7), indicating that JA and SA enhanced the defense ability of the plant to some extent in response to T. absoluta feeding. In addition, we determined the expression levels of JA- and SA-related genes and found that the expressions of AOS1, AOC, PAL1, and SAMT in the treatment group were significantly higher than in the control (Figure 8). The study demonstrated that tomato plants treated with B. amyloliquefaciens MBI600 significantly elevated the transcript levels of LoxD (JA-related) and NPR1 (SA-related) [53]. In N. benthamiana, B. amyloliquefaciens FZB42 colonization increased JA and SA contents in the leaves and upregulated the expression levels of LOX and PR-1a involved in the JA and SA signaling pathways [54]. Similarly in N. benthamiana, B. amyloliquefaciens KRS005 significantly increased the expressions of JA- and SA-related genes, including LOX, PR1, PR2, and PR4, in the leaves [41]. These studies implied that endophyte colonization might enhance plant resistance against insect pests by regulating JA- and SA-related gene expressions. However, the specific regulatory mechanisms require more in-depth research in future.

5. Conclusions

We found that B. amyloliquefaciens W10 treatment (2 × 108 CFU/mL twice) promoted tomato growth and led to a significant decrease in the fecundity of T. absoluta. The population growth simulation showed a negative effect on the population size after B. amyloliquefaciens W10 treatment (2 × 108 CFU/mL twice). Meanwhile, their oviposition preference was significantly reduced. The results also demonstrated that the mechanism of B. amyloliquefaciens W10 in controlling T. absoluta was the regulation of ROS levels and induction of defense-related enzymes. The content of JA and SA, as well as the expressions of their related genes, were significantly increased after B. amyloliquefaciens W10 treatment. This study clarified the ability of B. amyloliquefaciens W10 to resist T. absoluta for the first time and provided a new direction for research on the biological control of T. absoluta. However, further research is needed on the potential mechanisms of rhizobacterium-plant-insect interactions.

Supplementary Materials

The following are available online at https://www.mdpi.com/article/10.3390/agronomy15030695/s1, Table S1: Primers used in this study; Table S2: Biological parameters of T. absoluta fed on the tomato plants treated with B. amyloliquefaciens W10; Figure S1: Colonization verification of B. amyloliquefaciens W10 in the roots of tomato seedlings; Figure S2: Age-stage specific survival rate (sxj) of T. absoluta fed on the tomato plants treated with B. amyloliquefaciens W10; with B. amyloliquefaciens W10; Figure S3: Age-stage specific life expectancy (exj) of T. absoluta fed on the tomato plants treated with B. amyloliquefaciens W10.

Author Contributions

Conceptualization, G.X. and G.Y.; Software, M.Q., C.S. and G.X.; Formal analysis, M.Q., C.S., M.Z., K.Z. and Y.Y.; Investigation, M.Q., C.S., M.Z., K.Z. and Y.Y.; Data curation, C.S.; Writing—original draft, M.Q.; Writing—review and editing, G.X.; Visualization, C.S.; Supervision, G.Y.; Project administration, G.Y.; Funding acquisition, G.Y. All authors have read and agreed to the published version of the manuscript.

Funding

The National Key Research and Development Program of China (2021YFD1400200) supported this work.

Data Availability Statement

Data are contained within the article or Supplementary Material.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Desneux, N.; Wajnberg, E.; Wyckhuys, K.A.G.; Burgio, G.; Arpaia, S.; Narváez-Vasquez, C.A.; González-Cabrera, J.; Ruescas, D.C.; Tabone, E.; Frandon, J.; et al. Biological invasion of European tomato crops by Tuta absoluta: Ecology, geographic expansion and prospects for biological control. J. Pest Sci. 2010, 83, 197–215. [Google Scholar] [CrossRef]
  2. Jabamo, T.; Ayalew, G.; Goftishu, M.; Wakgari, M. Integrated effect of insecticide and sex pheromone on the tomato leafminer, Tuta absoluta (Lepidoptera: Gelechiidae). Crop Prot. 2023, 171, 106285. [Google Scholar] [CrossRef]
  3. Wang, M.H.; Ismoilov, K.; Liu, W.X.; Bai, M.; Bai, X.S.; Chen, B.; Chen, H.L.; Chen, H.S.; Dong, Y.C.; Fang, K.; et al. Tuta absoluta management in China: Progress and prospects. Entomol. Gen. 2024, 44, 269–278. [Google Scholar] [CrossRef]
  4. Hassan, E.M.; Mesbah, I.I.; Ali, F.A. Prevalence, population dynamics and associated natural enemies of Tomato Leafminer, Tuta absoluta, in Egypt. Int. J. Trop. Insect Sci. 2022, 42, 143–162. [Google Scholar] [CrossRef]
  5. Biondi, A.; Guedes, R.N.C.; Wan, F.H.; Desneux, N. Ecology, worldwide spread, and management of the invasive South American tomato pinworm, Tuta absoluta: Past, present, and future. Annu. Rev. Entomol. 2018, 63, 239–258. [Google Scholar] [CrossRef]
  6. Sohrabi, F.; Nooryazdan, H.R.; Gharati, B.; Saeidi, Z. Plant resistance to the moth Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) in tomato cultivars. Neotrop. Entomol. 2017, 46, 203–209. [Google Scholar] [CrossRef]
  7. Desneux, N.; Luna, M.G.; Guillemaud, T.; Urbaneja, A. The invasive South American tomato pinworm, Tuta absoluta, continues to spread in Afro-Eurasia and beyond: The new threat to tomato world production. J. Pest Sci. 2011, 84, 403–408. [Google Scholar] [CrossRef]
  8. Han, P.; Zhang, Y.B.; Arnó, J.; Mansour, R. Research toward enhancing integrated management of Tuta absoluta, an ongoing invasive threat in Afro-Eurasia. Entomol. Gen. 2024, 44, 263–267. [Google Scholar] [CrossRef]
  9. Aboutalebian-Soureshjani, A.; Rafiee-Dastjerdi, H.; Naseri, B.; Hassanpour, M.; Khajehali, J. Indoxacarb resistance in Iranian populations of Tuta absoluta (Lepidoptera: Gelechiidae): Cross-resistance, biochemical and molecular mechanisms. Pestic. Biochem. Physiol. 2023, 196, 105633. [Google Scholar] [CrossRef]
  10. Zang, L.S.; Akhtar, Z.R.; Ali, A.; Tairq, K.; Campos, M.R. Flubendiamide resistance and its mode of inheritance in tomato pinworm Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae). Insects 2022, 13, 1023. [Google Scholar] [CrossRef]
  11. Guedes, R.N.C.; Roditakis, E.; Campos, M.R.; Haddi, K.; Bielza, P.; Siqueira, H.A.A.; Tsagkarakou, A.; Vontas, J.; Nauen, R. Insecticide resistance in the tomato pinworm Tuta absoluta: Patterns, spread, mechanisms, management and outlook. J. Pest Sci. 2019, 92, 1329–1342. [Google Scholar] [CrossRef]
  12. Salazar-Mendoza, P.; Magalhes, D.M.; Pec, M.; Azevedo, K.E.X.; Delalibera, I.; Bento, J.M.S. Endophytic entomopathogenic fungus, individually and in combination with rhizobacteria, enhances resistance in wild and cultivated tomatoes to Tuta absoluta. J. Pest Sci. 2024. [Google Scholar] [CrossRef]
  13. Damme, V.M.V.; Beck, B.K.; Berckmoes, E.; Moerkens, R.; Wittemans, L.; De, V.R.; Nuyttens, D.; Casteels, H.F.; Maes, M.; Tirry, L.; et al. Efficacy of entomopathogenic nematodes against larvae of Tuta absoluta in the laboratory. Pest Manag. Sci. 2016, 72, 1702–1709. [Google Scholar] [CrossRef]
  14. Zhivko, M.; Beatriz, R.S.; Laura, D.; Ana, S.L.D.; Guadalupe, Z.H.; Dimitri, O.; Haymanti, S.; Dimitra, P.; Juan, M.G.; Alica, G.C.; et al. Beneficial soil fungi enhance tomato crop productivity and resistance to the leaf-mining pest Tuta absoluta in agronomic conditions. Agron. Sustain. Dev. 2024, 44, 55. [Google Scholar]
  15. Zhu, Y.X.; Zhang, Y.Y.; Wang, X.Y.; Yin, Y.; Du, Y.Z. Wolbachia modify host cell metabolite profiles in response to short-term temperature stress. Environ. Microbiol. Rep. 2024, 16, e70013. [Google Scholar] [CrossRef]
  16. Panwar, N.; Szczepaniec, A. Endophytic entomopathogenic fungi as biological control agents of insect pests. Pest Manag. Sci. 2024, 80, 6033–6040. [Google Scholar] [CrossRef]
  17. Chaudhary, P.; Agri, U.; Chaudhary, A.; Kumar, A.; Kumar, G. Endophytes and their potential in biotic stress management and crop production. Front. Microbiol. 2022, 13, 933017. [Google Scholar] [CrossRef]
  18. Grabka, R.; d’Entremont, T.W.; Adams, S.J.; Walker, A.K.; Tanney, J.B.; Abbasi, P.A.; Ali, S. Fungal endophytes and their role in agricultural plant protection against pests and pathogens. Plants 2022, 11, 384. [Google Scholar] [CrossRef]
  19. Zhang, J.M.; Huang, X.Q.; Yang, S.D.; Huang, A.R.; Ren, J.; Luo, X.G.; Feng, S.; Li, P.H.; Li, Z.G.; Dong, P.; et al. Endophytic Bacillus subtilis H17-16 effectively inhibits Phytophthora infestans, the pathogen of potato late blight, and its potential application. Pest Manag. Sci. 2023, 79, 5073–5086. [Google Scholar] [CrossRef]
  20. González-Cabrera, J.; Mollá, O.; Montón, H.; Urbaneja, A. Efficacy of Bacillus thuringiensis (Berliner) in controlling the tomato borer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae). Biocontrol 2011, 56, 71–80. [Google Scholar] [CrossRef]
  21. Li, H.Y.; Soares, M.A.; Torres, M.S.; Bergen, M.; White, J.F., Jr. Endophytic bacterium, Bacillus amyloliquefaciens, enhances ornamental hosta resistance to diseases and insect pests. J. Plant Interact. 2015, 10, 224–229. [Google Scholar] [CrossRef]
  22. Guo, S.; Pan, R.B.; Zhang, Y.; Gu, Q.W.; Shen, Q.R.; Yang, J.; Huang, L.Q.; Shen, Z.Z.; Li, R. Plant-microbe interactions influence plant performance via boosting beneficial root-endophytic bacteria. Environ. Microbiome 2025, 20, 18. [Google Scholar] [CrossRef] [PubMed]
  23. Miljakovic, D.; Marinkovic, J.; Balesevic-Tubic, S. The significance of Bacillus spp. In disease suppression and growth promotion of field and vegetable crops. Microorganisms 2020, 8, 1037. [Google Scholar] [CrossRef] [PubMed]
  24. Gray, E.J.; Smith, D.L. Intracellular and extracellular PGPR: Commonalities and distinctions in the plant-bacterium signaling processes. Soil. Biol. Biochem. 2005, 37, 395–412. [Google Scholar] [CrossRef]
  25. Do, Q.T. The endophytic bacterium Bacillus subtilis R8 as a prospective biocontrol agent for managing tea blister blight and enhancing tea yield. BioControl 2025. [Google Scholar] [CrossRef]
  26. Pieterse, C.M.; Zamioudis, C.; Berendsen, R.L.; Weller, D.M.; Wees, S.C.V.; Bakker, P.A. Induced systemic resistance by beneficial microbes. Annu. Rev. Phytopathol. 2014, 52, 347–375. [Google Scholar] [CrossRef]
  27. Koul, B.; Singh, S.; Dhanjal, D.S.; Singh, J. Plant Growth-Promoting Rhizobacteria (PGPRs): A Fruitful Resource; Springer: Singapore, 2019. [Google Scholar]
  28. Xing, Z.X.; Liu, D.; Luo, M.; Yang, Z.L.; Pang, W.Y.; Feng, Y.X.; Yan, J.N.; He, F.M.; Feng, X.; Yuan, Q.; et al. Analysis of the control effect of Bacillus amyloliquefaciens C4 wettable powder on potato bacterial wilt caused by Ralstonia solanacearum. Agronomy 2025, 15, 206. [Google Scholar] [CrossRef]
  29. Pires, L.M.; Marques, E.J.; Oliveira, J.V.; Alves, S.B. Selection of isolates of entomopathogenic fungi for controlling Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) and their compatibility with insecticides used in tomato crop. Neotrop. Entomol. 2010, 39, 977–984. [Google Scholar] [CrossRef]
  30. Liang, M.; Feng, A.; Wang, C.; Zhu, X.; Su, J.; Xu, Z.; Yang, J.; Wang, W.; Chen, K.; Chen, B.; et al. Bacillus amyloliquefaciens LM-1 affects multiple cell biological processes in Magnaporthe oryzae to suppress rice blast. Microorganisms 2024, 12, 1246. [Google Scholar] [CrossRef]
  31. Dimopoulou, A.; Theologidis, I.; Varympopi, A.; Papafotis, D.; Mermigka, G.; Tzima, A.; Panopoulos, N.J.; Skandalis, N. Shifting perspectives of translational research in bio-bactericides: Reviewing the Bacillus amyloliquefaciens paradigm. Biology 2021, 10, 1202. [Google Scholar] [CrossRef]
  32. Duan, Y.; Chen, R.; Zhang, R.; Jiang, W.; Chen, X.; Yin, C.; Mao, Z. Isolation, identification, and antibacterial mechanisms of Bacillus amyloliquefaciens QSB-6 and its effect on plant roots. Front. Microbiol. 2021, 12, 746799. [Google Scholar] [CrossRef] [PubMed]
  33. Zhao, P.; Quan, C.; Wang, Y.; Wang, J.; Fan, S. Bacillus amyloliquefaciens Q-426 as a potential biocontrol agent against Fusarium oxysporum f. sp. spinaciae. J. Basic. Microbiol. 2013, 54, 448–456. [Google Scholar] [CrossRef]
  34. Yang, P.; Yuan, P.; Liu, W.S.; Bernier, M.C.; Zhang, C.Q.; Adhikari, A.; Opiyo, S.O.; Zhao, L.J.; Banks, F.; Xia, Y.; et al. Plant growth promotion and plant disease suppression induced by Bacillus amyloliquefaciens strain GD4a. Plants 2024, 13, 672. [Google Scholar] [CrossRef]
  35. Luan, P.Y.; Yi, Y.J.; Huang, Y.F.; Cui, L.Q.; Hou, Z.P.; Zhu, L.J.; Liu, Y. Biocontrol potential and action mechanism of Bacillus amyloliquefaciens DB2 on Bipolaris sorokiniana. Front. Microbiol. 2023, 14, 1149363. [Google Scholar] [CrossRef]
  36. Khan, R.; He, P.B.; Chen, X.J.; He, P.F.; Ahmed, A.; Wu, Y.X.; Tang, G.W.; Tang, P.; Li, X.Y.; Munir, S.; et al. Bacillus endophytes for sustainable management of tomato spotted wilt virus and yield production. Pest Manag. Sci. 2024. [Google Scholar] [CrossRef]
  37. Uwaremwe, C.; Yue, L.; Wang, Y.; Tian, Y.; Zhao, X.; Liu, Y.; Zhou, Q.; Zhang, Y.B.; Wang, R.Y. An endophytic strain of Bacillus amyloliquefaciens suppresses Fusarium oxysporum infection of Chinese wolfberry by altering its rhizosphere bacterial community. Front. Microbiol. 2021, 12, 782523. [Google Scholar] [CrossRef]
  38. Zhao, K.B.; Ma, R.Y.; Cheng, M.; Guo, T.; Wu, W.J.; Song, Y.X.; Xu, H.; Tan, A.P.; Qin, B.F.; Wei, S.P.; et al. Isolation of Macrolactin A from a new Bacillus amyloliquefaciens and its aphicidal activity against Rhopalosiphum padi. Pest Manag. Sci. 2024. [Google Scholar] [CrossRef]
  39. Sellappan, R.; Thangavel, K. Bacillus amyloliquefaciens MZ895491: An endophytic bacterium as potential biocontrol agent in integrated pest management of fall armyworm in maize. Biocontrol Sci. Technol. 2024, 34, 1150–1164. [Google Scholar] [CrossRef]
  40. Qian, S.; Ahmed, A.; He, P.; He, P.; Munir, S.; Xia, M.; Tang, C.; Tang, P.; Wang, Z.; Khan, R.; et al. Bacillus amyloliquefaciens AK-12 helps rapeseed establish a protection against Brevicoryne brassicae. Int. J. Mol. Sci. 2023, 24, 15893. [Google Scholar] [CrossRef]
  41. Qi, H.Y.; Wang, D.; Han, D.F.; Song, J.; Ali, M.; Dai, X.F.; Zhang, X.J.; Chen, J.Y. Unlocking antagonistic potential of Bacillus amyloliquefaciens KRS005 to control gray mold. Front. Microbiol. 2023, 14, 1189354. [Google Scholar] [CrossRef]
  42. Ji, Z.L.; Tang, L.J.; Zhang, Q.X.; Xu, J.Y.; Chen, X.J.; Tong, Y.H. Isolation, purification and characterization of antifungal protein from Bacillus licheniformis W10 strain. Acta Phytophy. Sin. 2007, 3, 38–42. [Google Scholar]
  43. Xu, G.; Fu, L.R.; Wu, L.; Lu, J.; Xu, M.Q.; Qian, R.H.; Shao, C.J.; Qian, M.S.; Zhang, Y.Y.; Yang, G.Q. A tyramine receptor gene LsTAR2 is involved in reproduction and feeding in the small brown planthopper Laodelphax striatellus. Pestic. Biochem. Physiol. 2025, 209, 106335. [Google Scholar] [CrossRef]
  44. Chi, H. Life-table analysis incorporating both sexes and variable development rates among individuals. Environ. Entomol. 1988, 17, 26–34. [Google Scholar] [CrossRef]
  45. Chi, H.; Güncan, A.; Kavousi, A.; Gharakhani, G.; Atlihan, R.; Salih Özgökçe, M.; Shirazi, J.; Amir-Maafi, M.; Maroufpoor, M.; Roya, T. TWOSEX-MSChart: The key tool for life table research and education. Entomol. Gen. 2022, 42, 845–849. [Google Scholar] [CrossRef]
  46. Chi, H.; You, M.S.; Atlihan, R.; Smith, C.L.; Kavousi, A.; Ozgokce, M.S. Agestage, two-sex life table: An introduction to theory, data analysis, and application. Entomol. Gen. 2020, 40, 103–124. [Google Scholar] [CrossRef]
  47. Cao, Y.; Zhang, Z.W.; Xue, L.W.; Du, J.B.; Shang, J.; Xu, F.; Yuan, S.; Lin, H.H. Lack of salicylic acid in Arabidopsis protects plants against moderate salt stress. Z. Naturforsch. C 2009, 64, 231–238. [Google Scholar] [CrossRef]
  48. Imahori, Y.; Takemura, M.; Bai, J. Chilling-induced oxidative stress and antioxidant responses in mume (Prunus mume) fruit during low temperature storage. Postharvest Biol. Technol. 2008, 49, 54–60. [Google Scholar] [CrossRef]
  49. Zhang, L.; Oh, Y.; Li, H.Y.; Baldwin, I.T.; Galis, I. Alternative oxidase in resistance to biotic stresses: Nicotiana attenuata AOX contributes to resistance to a pathogen and a piercing-sucking insect but not Manduca sexta larvae. Plant Physiol. 2012, 160, 1453–1467. [Google Scholar] [CrossRef]
  50. Zhang, Y.Y.; Qian, M.S.; Shao, C.J.; Fu, L.R.; Wu, L.; Qian, R.H.; Xu, M.Q.; Lu, J.; Xu, G.; Yang, G.Q. Functional characterization of β-adrenergic-like octopamine receptors in planthopper reproduction and feeding. Int. J. Biol. Macromol. 2025, 288, 138722. [Google Scholar] [CrossRef]
  51. Livak, K.J.; Schmittgen, T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 2001, 25, 402–408. [Google Scholar] [CrossRef]
  52. Mittler, R.; Vanderauwera, S.; Gollery, M.; Breusegem, F.V. Reactive oxygen gene network of plants. Trends Plant Sci. 2004, 9, 490–498. [Google Scholar] [CrossRef] [PubMed]
  53. Beris, D.; Theologidis, I.; Skandalis, N.; Vassilakos, N. Bacillus amyloliquefaciens strain MBI600 induces salicylic acid dependent resistance in tomato plants against Tomato spotted wilt virus and Potato virus Y. Sci. Rep. 2018, 8, 10320. [Google Scholar] [CrossRef] [PubMed]
  54. Wu, L.M.; Huang, Z.Y.; Li, X.; Ma, L.M.; Gu, Q.; Wu, H.J.; Liu, J.; Borriss, R.; Wu, Z.; Gao, X.W. Stomatal closure and SA-, JA/ET-signaling pathways are essential for Bacillus amyloliquefaciens FZB42 to restrict leaf disease caused by Phytophthora nicotianae in Nicotiana benthamiana. Front. Microbiol. 2018, 9, 9847. [Google Scholar] [CrossRef] [PubMed]
  55. Zheng, Y.Q.; Liu, Y.L.; Zhang, J.Y.; Liu, X.L.; Ju, Z.G.; Shi, H.X.; Zhou, W.W. Dual role of endophytic entomopathogenic fungi: Induce plant growth and control tomato leafminer Phthorimaea absoluta. Pest Manag. Sci. 2023, 79, 4557–4568. [Google Scholar] [CrossRef]
  56. Chi, H.; Kavousi, A.; Gharekhani, G.; Atlihan, R.; Salih Özgökçe, M.; Güncan, A. Advances in theory, data analysis, and application of the age-stage, two-sex life table for demographic research, biological control, and pest management. Entomol. Gen. 2023, 43, 705–732. [Google Scholar] [CrossRef]
  57. Gadhave, K.; Gange, A. Plant-associated Bacillus spp. alter life-history traits of the specialist insect Brevicoryne brassicae L. Agr. Forest Entomol. 2016, 18, 35–42. [Google Scholar] [CrossRef]
  58. Huangfu, N.B.; Shang, J.; Guo, L.X.; Zhu, X.Z.; Zhang, K.X.; Niu, R.C.; Li, D.Y.; Gao, X.K.; Wang, L.; Ji, J.C.; et al. Life table analysis and RNA-Seq reveal hormesis and transgenerational effects of deltamethrin on Aphis gossypii. Pest Manag. Sci. 2025, 81, 477–489. [Google Scholar] [CrossRef]
  59. Huangfu, N.B.; Guo, L.X.; Shang, J.; Wang, L.; Zhang, K.X.; Li, D.Y.; Gao, X.K.; Zhu, X.Z.; Ji, J.C.; Luo, J.Y.; et al. Hormetic dose response induced by sublethal-dose sulfoxaflor leads to reproductive stimulation of Aphis gossypii. Pestic. Biochem. Physiol. 2024, 204, 106061. [Google Scholar] [CrossRef]
  60. Xu, G.; She, S.Y.; Gui, W.; Ma, C.; Zhang, Y.Y.; Qian, M.S.; Yang, G.Q. Seed priming of rice varieties with decoyinine improve their resistance against the brown planthopper Nilaparvata lugens. Agronomy 2023, 13, 72. [Google Scholar] [CrossRef]
  61. Xu, G.; Li, C.T.; Gui, W.; Xu, M.Q.; Lu, J.; Qian, M.S.; Zhang, Y.Y.; Yang, G.Q. Colonization of Piriformospora indica enhances rice resistance against the brown planthopper Nilaparvata lugens. Pest Manag. Sci. 2024, 80, 4386–4398. [Google Scholar] [CrossRef]
  62. Disi, J.; Simmons, J.; Zebelo, S. Plant growth-promoting rhizobacteria-induced defense against insect herbivores. In Field Crops: Sustainable Management by PGPR; Maheshwari, D.K., Dheeman, S., Eds.; Springer International Publishing: Cham, Switzerland, 2019; pp. 385–410. [Google Scholar]
  63. Bertea, C.M.; Casacci, L.P.; Bonelli, S.; Zampollo, A.; Barbero, F. Chemical, physiological and molecular responses of host plants to Lepidopteran egg-laying. Front. Plant Sci. 2020, 10, 1768. [Google Scholar] [CrossRef] [PubMed]
  64. Arturo, R.O.; Hexon Angel, C.C.; Nuvia, O.C.; Alfonso, L.C. Robert Winkler, Lourdes Macías-Rodríguez. Volatiles released by Beauveria bassiana induce oviposition behavior in the fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae). FEMS Microbiol. Ecol. 2022, 98, 114. [Google Scholar]
  65. Ramamoorthy, V.; Viswanathan, R.; Raguchander, T.; Prakasam, V.; Samiyappan, R. Induction of systemic resistance by plant growth promoting rhizobacteria in crop plants against pests and diseases. Crop Prot. 2001, 20, 1–11. [Google Scholar] [CrossRef]
  66. Chen, G.; Hu, W.Y.; Xie, F.D.; Zhang, L.J. Solvent for extracting malondialdehyde in plant as an index of senescence. Plant Physiol. J. 1991, 1, 44–46. [Google Scholar]
  67. Zhao, Z.C.; Yu, M.X.; Wei, Y.Y.; Xu, F.; Jiang, S.; Chen, Y.; Ding, P.; Shao, X.F. Cinnamon essential oil causes cell membrane rupture and oxidative damage of Rhizopus stolonifer to control soft rot of peaches. Food Control 2025, 170, 111039. [Google Scholar] [CrossRef]
  68. Yang, T.; Zhu, L.S.; Meng, Y.; Lv, R.; Zhou, Z.; Zhu, L.; Lin, H.H.; Xi, D.H. Alpha-momorcharin enhances Tobacco mosaic virus resistance in tobaccoNN by manipulating jasmonic acid-salicylic acid crosstalk. J. Plant Physiol. 2018, 223, 116–126. [Google Scholar] [CrossRef]
  69. Mukami, A.; Ngetich, A.; Mweu, C.; Oduor, R.O.; Muthangya, M.; Mbinda, W.M. Differential characterization of physiological and biochemical responses during drought stress in finger millet varieties. Physiol. Mol. Biol. Plants 2019, 25, 837–846. [Google Scholar] [CrossRef]
  70. Ji, Z.L.; Peng, S.; Zhu, W.; Dong, J.P.; Zhu, F. Induced resistance in nectarine fruit by Bacillus licheniformis W10 for the control of brown rot caused by Monilinia fructicola. Food Microbiol. 2020, 92, 103558. [Google Scholar] [CrossRef]
  71. Mhamdi, A.; Van Breusegem, F. Reactive oxygen species in plant development. Development 2018, 145, dev164376. [Google Scholar] [CrossRef]
  72. Mittler, R.; Zandalinas, S.I.; Fichman, Y.; Van, B.F. Reactive oxygen species signalling in plant stress responses. Nat. Rev. Mol. Cell Biol. 2022, 23, 663–679. [Google Scholar] [CrossRef]
  73. Cai, Y.Y.; Chen, J. Production and role of reactive oxygen species in plant defense responsesly. Chin. Bull. Bot. 1999, 16, 107–111. [Google Scholar]
  74. Deng, X.G.; Zhu, T.; Zou, L.J.; Han, X.Y.; Zhou, X.; Xi, D.H.; Zhang, D.W.; Lin, H.H. Orchestration of hydrogen peroxide and nitric oxide in brassinosteroid-mediated systemic virus resistance in Nicotiana benthamiana. Plant J. 2016, 85, 478–493. [Google Scholar] [CrossRef] [PubMed]
  75. Yang, X.R.; Yan, S.Y.; Li, Y.J.; Li, G.S.; Zhao, Y.J.; Sun, S.Q.; Su, J.; Cui, Z.Q.; Huo, J.F.; Sun, Y.; et al. Defense-related enzyme activities and metabolomic analysis reveal differentially accumulated metabolites and response pathways for sheath blight resistance in rice. Plants 2024, 13, 3554. [Google Scholar] [CrossRef]
  76. Poór, P.; Ördög, A.; Czékus, Z.; Borbély, P.; Takács, Z.; Kovács, J.; Tari, I. Regulation of the key antioxidant enzymes by developmental processes and environmental stresses in the dark. Biol. Plant. 2018, 62, 201–210. [Google Scholar] [CrossRef]
  77. Ren, A.T.; Zhu, Y.; Chen, Y.L.; Ren, H.X.; Li, J.Y.; Abbott, L.K.; Xiong, Y.C. Arbuscular mycorrhizal fungus alters root-sourced signal (abscisic acid) for better drought acclimation in Zea mays L. seedlings. Environ. Exp. Bot. 2019, 167, 103824. [Google Scholar] [CrossRef]
  78. Hu, S.S.; Hu, B.; Chen, Z.B.; Vosátka, M.; Vymazal, J. Antioxidant response in arbuscular mycorrhizal fungi inoculated wetland plant under Cr stress. Environ. Res. 2020, 191, 110203. [Google Scholar] [CrossRef]
  79. Kurniawan, A.; Chuang, H.W. Rhizobacterial Bacillus mycoides functions in stimulating the antioxidant defence system and multiple phytohormone signalling pathways to regulate plant growth and stress tolerance. J. Appl. Microbiol. 2022, 132, 1260–1274. [Google Scholar] [CrossRef]
Agronomy 15 00695 g001
Figure 1. Age-stage reproductive value (vxj) of T. absoluta fed on the tomato treated by B. amyloliquefaciens W10. (A) Control; (B) B. amyloliquefaciens W10 (2 × 108 CFU/mL twice).
Figure 1. Age-stage reproductive value (vxj) of T. absoluta fed on the tomato treated by B. amyloliquefaciens W10. (A) Control; (B) B. amyloliquefaciens W10 (2 × 108 CFU/mL twice).
Agronomy 15 00695 g001
Agronomy 15 00695 g002
Figure 2. Projection of population growth of T. absoluta fed on the tomato treated by B. amyloliquefaciens W10. (A) Control; (B) B. amyloliquefaciens W10 (2 × 108 CFU/mL twice).
Figure 2. Projection of population growth of T. absoluta fed on the tomato treated by B. amyloliquefaciens W10. (A) Control; (B) B. amyloliquefaciens W10 (2 × 108 CFU/mL twice).
Agronomy 15 00695 g002
Agronomy 15 00695 g003
Figure 3. Oviposition preference of T. absoluta on the tomato treated by B. amyloliquefaciens W10 (2 × 108 CFU/mL twice) on the fifth day. Asterisks indicate statistical differences between the control and treatment (*** p < 0.001). Data represent mean ± SE.
Figure 3. Oviposition preference of T. absoluta on the tomato treated by B. amyloliquefaciens W10 (2 × 108 CFU/mL twice) on the fifth day. Asterisks indicate statistical differences between the control and treatment (*** p < 0.001). Data represent mean ± SE.
Agronomy 15 00695 g003
Agronomy 15 00695 g004
Figure 4. Electrical conductivity (A) and MDA content (B) in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. Asterisks indicate statistical differences between the control and treatment (** p < 0.01; *** p < 0.001; ns, no significant difference). Data represent mean ± SE.
Figure 4. Electrical conductivity (A) and MDA content (B) in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. Asterisks indicate statistical differences between the control and treatment (** p < 0.01; *** p < 0.001; ns, no significant difference). Data represent mean ± SE.
Agronomy 15 00695 g004
Agronomy 15 00695 g005
Figure 5. Changes in H2O2 (A) and O2− (B) content in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. Asterisks indicate statistical differences between the control and treatment (* p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significant difference). Data represent mean ± SE.
Figure 5. Changes in H2O2 (A) and O2− (B) content in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. Asterisks indicate statistical differences between the control and treatment (* p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significant difference). Data represent mean ± SE.
Agronomy 15 00695 g005
Agronomy 15 00695 g006
Figure 6. The activities of the defense-related enzymes in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. (A) POD, peroxidase; (B) SOD, superoxide dismutase; (C) CAT, catalase; (D) GST, glutathione s-transferases; (E) PAL, phenylalnine ammonialyase; (F) PPO, polyphenol oxidase. Asterisks indicate statistical differences between the control and treatment (* p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significant difference). Data represent mean ± SE.
Figure 6. The activities of the defense-related enzymes in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. (A) POD, peroxidase; (B) SOD, superoxide dismutase; (C) CAT, catalase; (D) GST, glutathione s-transferases; (E) PAL, phenylalnine ammonialyase; (F) PPO, polyphenol oxidase. Asterisks indicate statistical differences between the control and treatment (* p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significant difference). Data represent mean ± SE.
Agronomy 15 00695 g006
Agronomy 15 00695 g007
Figure 7. The content of JA (A) and SA (B) in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. Asterisks indicate statistical differences between the control and treatment (* p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significant difference). Significant differences (p < 0.05) among the control groups or B. amyloliquefaciens W10-treated groups are denoted by different lowercase letters or uppercase letters, respectively. Data represent mean ± SE.
Figure 7. The content of JA (A) and SA (B) in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. Asterisks indicate statistical differences between the control and treatment (* p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significant difference). Significant differences (p < 0.05) among the control groups or B. amyloliquefaciens W10-treated groups are denoted by different lowercase letters or uppercase letters, respectively. Data represent mean ± SE.
Agronomy 15 00695 g007
Agronomy 15 00695 g008
Figure 8. Relative expression levels of JA- and SA-related genes in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. (A) AOS1, allene oxide synthase 1; (B) AOS2, allene oxide synthase 2; (C) AOC, allene oxide cyclase; (D) PAL1, phenylalanine ammonia-lyase 1; (E) SABP2, salicylic acid-binding protein 2; (F) SAMT, salicylic acid methyltransferase. Asterisks indicate statistical differences between the control and treatment (* p < 0.05; *** p < 0.001; ns, no significant difference). Data represent mean ± SE.
Figure 8. Relative expression levels of JA- and SA-related genes in the control and B. amyloliquefaciens W10-treated (2 × 108 CFU/mL twice) tomato leaves after T. absoluta infestation. (A) AOS1, allene oxide synthase 1; (B) AOS2, allene oxide synthase 2; (C) AOC, allene oxide cyclase; (D) PAL1, phenylalanine ammonia-lyase 1; (E) SABP2, salicylic acid-binding protein 2; (F) SAMT, salicylic acid methyltransferase. Asterisks indicate statistical differences between the control and treatment (* p < 0.05; *** p < 0.001; ns, no significant difference). Data represent mean ± SE.
Agronomy 15 00695 g008
Table 1. The growth of tomato seedlings treated by different concentrations of B. amyloliquefaciens W10.
Table 1. The growth of tomato seedlings treated by different concentrations of B. amyloliquefaciens W10.
TreatmentStem Diameter/cmStem Height/cmShoot Fresh Weight/gRoot Fresh Weight/g
Control0.31 ± 0.01 a27.88 ± 0.92 c7.68 ± 0.31 b1.41 ± 0.11 d
108 CFU/mL (once)0.28 ± 0.01 a29.68 ± 1.01 bc8.20 ± 0.27 ab1.58 ± 0.09 cd
2 × 108 CFU/mL (once)0.30 ± 0.01 a30.02 ± 0.88 bc8.12 ± 0.28 ab1.74 ± 0.10 bcd
4 × 108 CFU/mL (once)0.30 ± 0.01 a31.31 ± 0.95 abc8.44 ± 0.34 ab2.16 ± 0.11 abc
108 CFU/mL (twice)0.29 ± 0.01 a29.31 ± 1.02 bc8.81 ± 0.35 ab2.26 ± 0.19 ab
2 × 108 CFU/mL (twice)0.29 ± 0.01 a32.98 ± 1.08 ab9.42 ± 0.22 a2.41 ± 0.14 a
4 × 108 CFU/mL (twice)0.32 ± 0.01 a35.24 ± 0.84 a9.40 ± 0.44 a2.40 ± 0.19 a
Means ± SE followed by different lowercase letters within a column are significantly different among different B. amyloliquefaciens W10 treatments (p < 0.05, Tukey’s HSD test).
Table 2. The survival rate and fecundity of T. absoluta fed on the tomato treated by different concentrations of B. amyloliquefaciens W10.
Table 2. The survival rate and fecundity of T. absoluta fed on the tomato treated by different concentrations of B. amyloliquefaciens W10.
TreatmentLarval StagePupal StagePre-Mature StageFecundity (Eggs)
Control0.91 ± 0.01 a0.86 ± 0.01 a0.78 ± 0.01 a124.25 ± 12.85 a
108 CFU/mL (once)0.91 ± 0.01 a0.85 ± 0.01 a0.78 ± 0.02 a126.67 ± 9.71 a
2 × 108 CFU/mL (once)0.90 ± 0.01 a0.87 ± 0.01 a0.79 ± 0.02 a95.64 ± 8.59 ab
4 × 108 CFU/mL (once)0.90 ± 0.01 a0.87 ± 0.01 a0.78 ± 0.02 a110.23 ± 12.88 ab
108 CFU/mL (twice)0.91 ± 0.01 a0.85 ± 0.01 a0.77 ± 0.02 a122.26 ± 10.79 ab
2 × 108 CFU/mL (twice)0.91 ± 0.01 a0.86 ± 0.01 a0.78 ± 0.02 a79.79 ± 6.50 b
4 × 108 CFU/mL (twice)0.89 ± 0.01 a0.87 ± 0.01 a0.77 ± 0.01 a87.78 ± 7.96 ab
The pre-mature stage refers to the overall survival rate from larvae to pupae; Means ± SE followed by different lowercase letters within a column are significantly different among different B. amyloliquefaciens W10 treatments (p < 0.05, Tukey’s HSD test).
Table 3. Population life table parameters of T. absoluta fed on the tomato treated by B. amyloliquefaciens W10.
Table 3. Population life table parameters of T. absoluta fed on the tomato treated by B. amyloliquefaciens W10.
ParametersControlW10 (2 × 108 CFU/mL)p
Net reproductive rate R0 (offspring)67.017 ± 10.68741.898 ± 7.0120.049
Intrinsic rate of increase rm (days−1)0.172 ± 0.0080.148 ± 0.0080.037
Mean generation time T (days)24.500 ± 0.43025.148 ± 0.4600.304
Finite capacity of increase λ (days−1)1.187 ± 0.0091.160 ± 0.0090.036
Doubling time DT (days)4.039 ± 0.1894.667 ± 0.2540.046
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Qian, M.; Sheng, C.; Zheng, M.; Zhu, K.; Yu, Y.; Xu, G.; Yang, G. A Bacterial Endophyte Bacillus amyloliquefaciens W10 Enhances the Tomato Resistance Against Tuta absoluta. Agronomy 2025, 15, 695. https://doi.org/10.3390/agronomy15030695

AMA Style

Qian M, Sheng C, Zheng M, Zhu K, Yu Y, Xu G, Yang G. A Bacterial Endophyte Bacillus amyloliquefaciens W10 Enhances the Tomato Resistance Against Tuta absoluta. Agronomy. 2025; 15(3):695. https://doi.org/10.3390/agronomy15030695

Chicago/Turabian Style

Qian, Mingshi, Chaoqi Sheng, Mingying Zheng, Ke Zhu, Youxin Yu, Gang Xu, and Guoqing Yang. 2025. "A Bacterial Endophyte Bacillus amyloliquefaciens W10 Enhances the Tomato Resistance Against Tuta absoluta" Agronomy 15, no. 3: 695. https://doi.org/10.3390/agronomy15030695

APA Style

Qian, M., Sheng, C., Zheng, M., Zhu, K., Yu, Y., Xu, G., & Yang, G. (2025). A Bacterial Endophyte Bacillus amyloliquefaciens W10 Enhances the Tomato Resistance Against Tuta absoluta. Agronomy, 15(3), 695. https://doi.org/10.3390/agronomy15030695

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop