Apple-arbuscular mycorrhizal symbiosis confers resistance to Fusarium solani by inducing defense response and elevating nitrogen absorption.

Fusarium solani exerts detrimental effects on plant growth, which is one of the reasons for the incidence of apple replant disease. Arbuscular mycorrhizal fungi (AMF) enhance plant resistance to Fusarium wilt; however, the mechanism remains poorly understood. Therefore, the present study investigated the symbiosis between apple and AMF and explored the physiology, especially nitrate metabolism, antioxidant defense, and photosynthetic performance, when infected by F. solani. The experiment was carried out with four treatments, namely -AMF - F. solani, -AMF + F. solani, -AMF + F. solani, and + AMF + F. solani. In this study, the -AMF + F. solani treatment increased the activity of enzymes associated with nitrogen metabolism, such as the nitrate and nitrite reductases, in the apple root system. The +AMF + F. solani treatment showed higher antioxidant enzyme activities than the -AMF + F. solani by F. solani infection. The apple seedlings of the +AMF + F. solani treatment decreased reactive oxygen accumulation and reduced the oxidative damages triggered by F. solani infection. The improvement in antioxidant capacity due to the +AMF + F. solani treatment was closely associated with the upregulation of genes related to the antioxidant system. The F. solani infection greatly damaged the photosynthetic process, while the +AMF + F. solani treatment significantly improved it compared to the -AMF + F. solani treatment. In conclusion, the study demonstrated that the apple-AMF symbiosis plays an active role in regulating the resistance against F. solani infection by enhancing defense response and nitrogen metabolism.

Remediation of the microecological environment of heavy metal-contaminated soil with fulvic acid, improves the quality and yield of apple.

The excessive application of chemical fertilizers and pesticides in apple orchards is responsible for high levels of manganese and copper in soil, and this poses a serious threat to soil health. We conducted a three-year field experiment to study the remediation effect and mechanism of fulvic acid on soil with excess manganese and copper. The exogenous application of fulvic acid significantly reduced the content of manganese and copper in soil and plants; increased the content of calcium; promoted the growth of apple plants; improved the fruit quality and yield of apple; increased the content of chlorophyll; increased the activity of superoxide dismutase, peroxidase, and catalase; and reduced the content of malondialdehyde. The number of soil culturable microorganisms, soil enzyme activity, soil microbial community diversity, and relative abundance of functional bacteria were increased, and the detoxification of the glutathione metabolism function was enhanced. The results of this study provide new insights that will aid the remediation of soil with excess manganese and copper using fulvic acid.

Effect of Hydrogen Peroxide on the Soil Microbial Community Structure and Growth of Malus hupehensis Rehd. Seedlings under Replant Conditions.

Apple replant disease (ARD) is common in apple production, which seriously affects the growth and development of apples. In this study, hydrogen peroxide with a bactericidal effect was used to treat the replanted soil, and the effects of different concentrations of hydrogen peroxide on replanted seedlings and soil microbiology were investigated in order to seek a green, clean way to control ARD. Five treatments were set up in this study: replanted soil (CK1), replanted soil with methyl bromide fumigation (CK2), replanted soil + 1.5% hydrogen peroxide (H1), replanted soil + 3.0% hydrogen peroxide (H2), and replanted soil + 4.5% hydrogen peroxide (H3). The results showed that hydrogen peroxide treatment improved replanted seedling growth and also inactivated a certain number of Fusarium, while the Bacillus, Mortierella, and Guehomyces also became more abundant in relative terms. The best results were obtained with replanted soil + 4.5% hydrogen peroxide (H3). Consequently, hydrogen peroxide applied to the soil can effectively prevent and control ARD.

Allium fistulosum L. Alleviates Apple Replant Disease by Suppressing Fusarium solani.

Fusarium solani has often been isolated from replanted apple roots, suggesting that it is associated with apple replant disease. The mechanism underlying the ability of the mixed cropping of apple trees with Allium fistulosum L. to alleviate apple replant disease remains unclear. The aim of this study was to determine the pathogenicity of the Fusarium solani isolate HBH 08 isolated from diseased roots and the effect of A. fistulosum L. and its root secretions on Fusarium solani isolate HBH 08 and apple seedings. The field experiment showed that A. fistulosum L. not only significantly reduced the amount of the Fusarium solani isolate HBH 08 in replanted soil but also increased the biomass of the grafted apple seedlings. The GC-MS analysis indicated that dimethyl disulphide and diallyl disulphide were active molecules in the root exudates of A. fistulosum L. They inhibited the growth of the Fusarium solani isolate HBH 08 mycelium and decreased the number of spores germinated. In addition, these compounds reduced the amount of the Fusarium solani isolate HBH 08 under replanted conditions and promoted the growth of grafted apple seedlings. Overall, mixed cropping with A. fistulosum L. might be an effective approach for cultivating apple trees and controlling apple replant disease.

Involvement of MdWRKY40 in the defense of mycorrhizal apple against fusarium solani.

BACKGROUND: Apple (Malus domestica Borkh.) is an important economic crop. The pathological effects of Fusarium solani, a species complex of soilborne pathogens, on the root systems of apple plants was unknown. It was unclear how mycorrhizal apple seedlings resist infection by F. solani. The transcriptional profiles of mycorrhizal and non-mycorrhizal plants infected by F. solani were compared using RNA-Seq. RESULTS: Infection with F. solani significantly reduced the dry weight of apple roots, and the roots of mycorrhizal apple plants were less damaged when the plants were infected with F. solani. They also had enhanced activity of antioxidant enzymes and a reduction in the oxidation of membrane lipids. A total of 1839 differentially expressed genes (DEGs) were obtained after mycorrhizal and non-mycorrhizal apple plants were infected with F. solani. A gene ontogeny (GO) analysis showed that most of the DEGs were involved in the binding of ADP and calcium ions. In addition, based on a MapMan analysis, a large number of DEGs were found to be involved in the response of mycorrhizal plants to stress. Among them, the overexpressed transcription factor MdWRKY40 significantly improved the resistance of the apple 'Orin' callus to F. solani and the expression of the resistance gene MdGLU by binding the promoter of MdGLU. CONCLUSION: This paper outlines how the inoculation of apple seedlings roots by arbuscular mycorrhizal fungi responded to infection with F. solani at the transcriptional level. In addition, MdWRKY40 played an important role in the resistance of mycorrhizal apple seedlings to infection with F. solani.

Chemical fumigants control apple replant disease: Microbial community structure-mediated inhibition of Fusarium and degradation of phenolic acids.

Fusarium and phenolic acids in apple replant soil have deleterious effects on soil, which affects the growth of young replanted apple trees. Here, we studied the effects of different chemical fumigants (metham sodium, dazomet, calcium cyanamide, 1,3-dichloropropene, and methyl bromide) on Fusarium and phenolic acids in soil. The chemical fumigants disturbed the apple replant soil microbial community to different degrees in the order from highest to the lowest as methyl bromide > 1,3-dichloropropene > dazomet > metham sodium > calcium cyanamide. Compared with the control, the total numbers of Operational Taxonomic Unit (OTU) were 104.63 % and 9.38 % lower in the methyl bromide and calcium cyanamide treatments, respectively while the average contents of Fusarium were 88.04 % and 59.18% lower in these treatments, respectively. Higher disturbance degrees resulted in a slower recovery rate of the soil microbial community, which facilitated the transformation of the soil into a disease-suppressing state. During the recovery process, the roots recruited Streptomyces OTU2796 and Bacillus OTU2243, which alleviated Fusarium-induced stress via the synthesis of polyketones and macrolides. The roots also recruited Sphingomonas OTU3488, OTU5572, and OTU8147, which alleviated phenolic acid-induced stress through the degradation of benzoate and polycyclic aromatic hydrocarbons.

Controlled-Release Diammonium Phosphate Alleviates Apple Replant Disease: An Integrated Analysis of Soil Properties, Plant Growth, and the Soil Microbiome.

Exogenous application of nitrogen (N) and phosphate (P) has been demonstrated to alleviate apple replant disease (ARD). Yet, the effect of controlled-release diammonium phosphate (C-DAP), which continuously supply N and P for ARD control, is still poorly understood. Applying C-DAP markedly alleviated the typical symptoms of ARD. C-DAP maintained soil N and P at relatively high and stable levels during the entire growth period of the replanted seedlings, thus, limiting the copy number of the four key pathogenic Fusarium species that cause ARD. Particularly, continuously supplying N and P by C-DAP established a higher fungal diversity than that of conventional diammonium phosphate and induced the fungal community to be more similar to fumigated soil. The positive effect of C-DAP originated from the synergistic effects of regulating microorganisms and enhancing the resistance of the plant caused by a continuous nutrient supply. These findings provide a new perspective in the management of soil-borne diseases.

Transcriptome changes associated with apple (Malus domestica) root defense response after Fusarium proliferatum f. sp. malus domestica infection.

BACKGROUND: Apple replant disease is a soilborne disease caused by Fusarium proliferatum f. sp. malus domestica strain MR5 (abbreviated hereafter as Fpmd MR5) in China. This pathogen causes root tissue rot and wilting leaves in apple seedlings, leading to plant death. A comparative transcriptome analysis was conducted using the Illumina Novaseq platform to identify the molecular defense mechanisms of the susceptible M.26 and the resistant M9T337 apple rootstocks to Fpmd MR5 infection. RESULTS: Approximately 518.1 million high-quality reads were generated using RNA sequencing (RNA-seq). Comparative analysis between the mock-inoculated and Fpmd MR5 infected apple rootstocks revealed 28,196 significantly differentially expressed genes (DEGs), including 14,572 up-regulated and 13,624 down-regulated genes. Among them, the transcriptomes in the roots of the susceptible genotype M.26 were reflected by overrepresented DEGs. MapMan analysis indicated that a large number of DEGs were involved in the response of apple plants to Fpmd MR5 stress. The important functional groups identified via gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment were responsible for fundamental biological regulation, secondary metabolism, plant-pathogen recognition, and plant hormone signal transduction (ethylene and jasmonate). Furthermore, the expression of 33 up-regulated candidate genes (12 related to WRKY DNA-binding proteins, one encoding endochitinase, two encoding beta-glucosidases, ten related to pathogenesis-related proteins, and eight encoding ethylene-responsive transcription factors) were validated by quantitative real-time PCR. CONCLUSION: RNA-seq profiling was performed for the first time to analyze response of apple root to Fpmd MR5 infection. We found that the production of antimicrobial compounds and antioxidants enhanced plant resistance to pathogens, and pathogenesis-related protein (PR10 homologs, chitinase, and beta-glucosidase) may play unique roles in the defense response. These results provide new insights into the mechanisms of the apple root response to Fpmd MR5 infection.

The Phlorizin-Degrading Bacillus licheniformis XNRB-3 Mediates Soil Microorganisms to Alleviate Apple Replant Disease.

In this study, an endophytic phlorizin-degrading Bacillus licheniformis XNRB-3 was isolated from the root tissue of healthy apple trees, and its control effect on apple replant disease (ARD) and how it alleviates the pathogen pressure via changes in soil microbiomes were studied. The addition of strain XNRB-3 in Fusarium infested soils significantly reduced the number of pathogens in the soil, thus resulting in a lower disease incidence, and the relative control effect on Fusarium oxysporum reached the highest of 66.11%. The fermentation broth can also protect the roots of the plants from Fusarium oxysporum, Fusarium moniliforme, Fusarium proliferatum, and Fusarium solani infection. These antagonistic effects were further validated using an in vitro assay in which the pathogen control was related to growth and spore germination inhibition via directly secreted antimicrobial substances and indirectly affecting the growth of pathogens. The secreted antimicrobial substances were identified using gas chromatography-mass spectrometry (GC-MS) technology. Among them, alpha-bisabolol and 2,4-di-tert-butylphenol had significant inhibitory effects on many planted pathogenic fungi. Butanedioic acid, monomethyl ester, and dibutyl phthalate promoted root development of Arabidopsis plants. Strain XNRB-3 has multifarious plant growth promoting traits and antagonistic potential. In pot and field experiments, the addition of strain XNRB-3 significantly promoted the growth of plants, and the activity of enzymes related to disease resistance [superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)] was also significantly enhanced. It also reduced the abundance of four species of Fusarium and the content of phenolic acids in the rhizosphere soil, improved soil microbial community structure and nutritional conditions, and increased soil microbial diversity and activity, as well as the soil enzyme activity. The above results indicated that B. licheniformis XNRB-3 could be developed into a promising biocontrol and plant-growth-promoting agent.

Quicklime and Superphosphate Alleviating Apple Replant Disease by Improving Acidified Soil.

A two-year field experiment was carried out in order to study the effect of different soil modifiers on alleviating apple replant disease (ARD) in the apple orchards. Four treatments were as follows: replanted apple orchard soil (CK), replanted apple orchard soil treated with quicklime 1.0 g·kg-1 (T1), replanted apple orchard soil treated with 1.0 g·kg-1 quicklime and 1.0 g·kg-1 superphosphate (T2), and replanted apple orchard soil treated with 1.0 g·kg-1 plant ash (T3). Soil pH, plant biomass, soil bacteria, soil fungi, Fusarium oxysporum, soil enzymes, plant chlorophyll, and photosynthetic parameters were measured to detect the improvement effects of different soil amendments on acidified soil and to alleviate the ARD. The three treatments stably raised the pH of acidified soil and improved the conditions of the plant rhizosphere environment. Compared with the control, T1, T2, and T3 treatments significantly increased growth and plant biomass indexes, such as plant height and ground diameter, as well as photosynthetic parameters. Among the three treatments, T2 had the strongest effects. In July 2018 and July 2019, the number of bacteria was 151.3 and 190.5% higher in T2-treated soil than in control soil, and the number of soil fungi was 53.6 and 53.3% lower. In 2018 and 2019, the copy number of Fusarium solani was 63.6 and 58.6% lower and that of F. oxysporum was 51.8 and 55.7% lower. The T1, T2, and T3 treatments significantly increased soil enzyme activity and leaf chlorophyll content, and their effects were generally ranked T2 > T1 > T3. In conclusion, a combination of 1.0 g·kg-1 quicklime and 1.0 g·kg-1 superphosphate added to acidified replant soil increased the soil pH, improved the soil environment, increased the number of bacteria, reduced the number of fungi, increased soil enzyme activity, and improved plant photosynthetic capacity, thereby promoting the growth of replanted seedlings and effectively reducing ARD.

Effects of Trichoderma asperellum 6S-2 on Apple Tree Growth and Replanted Soil Microbial Environment.

Trichoderma asperellum strain 6S-2 with biocontrol effects and potential growth-promoting properties was made into a fungal fertilizer for the prevention of apple replant disease (ARD). 6S-2 fertilizer not only promoted the growth of Malus hupehensis Rehd seedlings in greenhouse and pot experiments, but also increased the branch elongation growth of young apple trees. The soil microbial community structure changed significantly after the application of 6S-2 fertilizer: the relative abundance of Trichoderma increased significantly, the relative abundance of Fusarium (especially the gene copy numbers of four Fusarium species) and Cryptococcus decreased, and the relative abundance of Bacillus and Streptomyces increased. The bacteria/fungi and soil enzyme activities increased significantly after the application of 6S-2 fertilizer. The relative contents of alkenes, ethyl ethers, and citrullines increased in root exudates of M. hupehensis Rehd treated with 6S-2 fertilizer and were positively correlated with the abundance of Trichoderma. The relative contents of aldehydes, nitriles, and naphthalenes decreased, and they were positively correlated with the relative abundance of Fusarium. In addition, levels of ammonium nitrogen (NH4-N), nitrate nitrogen (NO3-N), available phosphorus (AP), available potassium (AK), organic matter (SOM), and pH in rhizosphere soil were also significantly related to changes in the microbial community structure. In summary, the application of 6S-2 fertilizer was effective in alleviating some aspects of ARD by promoting plant growth and optimizing the soil microbial community structure.

An emerging chemical fumigant: two-sided effects of dazomet on soil microbial environment and plant response.

Methyl bromide has been banned worldwide because it causes damage to the ozone layer and the environment. To find a substitute for methyl bromide, the relationships among fumigation, plant growth, and the microbial community in replant soil require further study. We performed pot and field experiments to investigate the effects of dazomet fumigation on soil properties and plant performance. Changes in soil microbial community structure and diversity were assessed using high-throughput sequencing, and plant physiological performance and soil physicochemical properties were also measured. Dazomet fumigation enhanced photosynthesis and promoted plant growth in replant soil; it altered soil physical and chemical properties and reduced soil enzyme activities, although these parameters gradually recovered over time. After dazomet fumigation, the dominant soil phyla changed, microbial diversity decreased significantly, the relative abundance of biocontrol bacteria such as Mortierella increased, and the relative abundance of pathogenic bacteria such as Fusarium decreased. Over the course of the experiment, the soil microbial flora changed dynamically, and soil enzyme activities and other physical and chemical properties also recovered to a certain extent. This result suggested that the effect of dazomet on soil microorganisms was temporary. However, fumigation also led to an increase in some resistant pathogens, such as Trichosporon, that affect soil function and health. Therefore, it is necessary to consider potential negative impacts of dazomet on the soil environment and to perform active environmental risk management in China.

The Endophytic Strain Trichoderma asperellum 6S-2: An Efficient Biocontrol Agent against Apple Replant Disease in China and a Potential Plant-Growth-Promoting Fungus.

A study was conducted for endophytic antagonistic fungi obtained from the roots of healthy apple trees growing in nine replanted orchards in Shandong Province, China. The fungi were assessed for their ability to inhibit Fusarium proliferatum f. sp. malus domestica MR5, a fungal strain associated with apple replant disease (ARD). An effective endophyte, designated as strain 6S-2, was isolated and identified as Trichoderma asperellum. Strain 6S-2 demonstrated protease, amylase, cellulase, and laccase activities, which are important for the parasitic and antagonistic functions of pathogenic fungi. The inhibition rate of 6S-2 against Fusarium proliferatum f. sp. malus domestica MR5 was 52.41%. Strain 6S-2 also secreted iron carriers, auxin, ammonia and was able to solubilize phosphorus. Its fermentation extract and volatile substances inhibited the growth of MR5, causing its hyphae to twist, shrink, swell, and rupture. The antifungal activity of the 6S-2 fermentation extract increased with increasing concentrations. It promoted the production and elongation of Arabidopsis thaliana lateral roots, and the strongest effects were seen at a concentration of 50 mg/mL. A GC-MS analysis of the 6S-2 fermentation extract and volatile substances showed that they comprised mainly alkanes, alcohols, and furanones, as well as the specific volatile substance 6-PP. The application of 6S-2 spore suspension to replanted apple orchard soils reduced plant oxidative damage and promoted plant growth in a pot experiment. Therefore, the endophytic strain T. asperellum 6S-2 has the potential to serve as an effective biocontrol fungus for the prevention of ARD in China, and appears to promote plant growth.

Isolation, Identification, and Antibacterial Mechanisms of Bacillus amyloliquefaciens QSB-6 and Its Effect on Plant Roots.

Apple replant disease (ARD) is a common problem in major apple planting areas, and biological factors play a leading role in its etiology. Here, we isolated the bacterial strain QSB-6 from the rhizosphere soil of healthy apple trees in a replanted orchard using the serial dilution method. Strain QSB-6 was provisionally identified as Bacillus amyloliquefaciens based on its morphology, physiological and biochemical characteristics, carbon source utilization, and chemical sensitivity. Maximum likelihood analysis based on four gene sequences [16S ribosomal RNA gene (16S rDNA), DNA gyrase subunit A (gyrA), DNA gyrase subunit B (gyrB), and RNA polymerase subunit B (rpoB)] from QSB-6 and other strains indicated that it had 100% homology with B. amyloliquefaciens, thereby confirming its identification. Flat standoff tests showed that strain QSB-6 had a strong inhibitory effect on Fusarium proliferatum, Fusarium solani, Fusarium verticillioides, Fusarium oxysporum, Alternaria alternata, Aspergillus flavus, Phoma sp., Valsa mali, Rhizoctonia solani, Penicillium brasilianum, and Albifimbria verrucaria, and it had broad-spectrum antibacterial characteristics. Extracellular metabolites from strain QSB-6 showed a strong inhibitory effect on Fusarium hyphal growth and spore germination, causing irregular swelling, atrophy, rupture, and cytoplasmic leakage of fungal hyphae. Analysis of its metabolites showed that 1,2-benzenedicarboxylic acid and benzeneacetic acid, 3- hydroxy-, methyl ester had good inhibitory effects on Fusarium, and increased the length of primary roots and the number of lateral roots of Arabidopsis thaliana plantlet. Pot experiments demonstrated that a QSB-6 bacterial fertilizer treatment (T2) significantly improved the growth of Malus hupehensis Rehd. seedlings. It increased root length, surface area, tips, and forks, respiration rate, protective enzyme activities, and the number of soil bacteria while reducing the number of soil fungi. Fermentation broth from strain QSB-6 effectively prevented root damage from Fusarium. terminal restriction fragment length polymorphism (T-RFLP) and quantitative PCR (qPCR) assays showed that the T2 treatment significantly reduced the abundance of Fusarium in the soil and altered the soil fungal community structure. In summary, B. amyloliquefaciens QSB-6 has a good inhibitory effect on Fusarium in the soil and can significantly promote plant root growth. It has great potential as a biological control agent against ARD.

Comprehensive Analysis of the Influence of Fulvic Acid from Paper Mill Effluent on Soil Properties, Soil Microbiome, and Growth of Malus hupehensis Rehd. Seedlings under Replant Conditions.

In this study, the potential regulatory effects of fulvic acid extracted from paper mill effluent (PFA) in apple replant disease (ARD) were investigated through a comprehensive experimental evaluation of the effects of PFA on soil properties, growth inhibition of apple replant pathogens, and growth of replanted Malus hupehensis Rehd. seedlings. PFA with a relatively lower molecular weight was mainly composed of carbohydrates, lignin derivatives, and polysaccharides and was rich in functional groups such as carboxyl and phenolic hydroxyl groups. Treatment with PFA dosages ranging from 2 to 3 g/pot significantly increased available phosphorus (P) in soil by 47.5 to 57.5% when compared with the control without PFA, indicating that PFA had a positive effect in activating P. In addition, PFA stimulated the growth of replanted seedlings by promoting root elongation, enhancing leaf photosynthesis, and increasing the activity of root antioxidant enzymes including superoxide dismutase, peroxidase, and catalase. However, no convincing evidence was found that application of different dosages of PFA had remarkable effects on soil pH, inorganic nitrogen, available potassium, organic matter, and the numbers of bacteria and fungi. Notably, PFA had no effect on the copy number of the main pathogenic fungi causing ARD, including Fusarium oxysporum, Fusarium solani, Fusarium proliferatum, and Fusarium moniliforme. Overall, PFA can alleviate ARD to a certain extent mainly through its effects on improving the resilience of replanted young seedlings rather than by affecting soil microorganisms or providing nutrients.

Key indicators for renewal and reconstruction of perennial trees soil: Microorganisms and phloridzin.

Perennial tree soil inhibits the growth of replanting apples, but the mechanism that underlies this inhibition is poorly understood. A total of 57 perennial tree soils were selected for the collection of soil samples in the Bohai Bay in May 2018. The severity of apple replant disease (ARD) for each soil was determined by calculating the rate of inhibition of growth replanted apple trees. A high-throughput sequencing analysis of internal transcribed spacer (ITS) was used to determine the soil fungal community. A correlation analysis was used to determine the relationship between the rate of inhibition of apple growth and soil factors. The degree of inhibition of plant growth varied substantially among the 57 soil samples examined. Different perennial tree soils have varying degrees of ARD. There was no significant difference in the composition of fungal community at the phylum level, but the genus level differed substantially. The abundances of Fusarium and Mortierella species and the contents of phloridin in the soil and soil organic matter (SOM) were significantly correlated with ARD severity. Structural equation modeling also emphasized that the degree of occurrence of ARD was directly or indirectly affected by Fusarium, Mortierella, phloridin and SOM. A correlation analysis can only be used as an indicator, and further research is merited to reveal how soil parameters affect ARD.

Effect of Emerging Soil Chemical Amendments on the Replant Soil Environment and Growth of Malus hupehensis Rehd. Seedlings.

The effects of different soil chemical amendments (T1, 1‰ quicklime + 1‰ superphosphate; T2, 1‰ quicklime; T3, 1‰ superphosphate) on the soil environment and growth of Malus hupehensis Rehd. seedlings in aged apple orchard soil were studied to provide new insight into the prevention and control of apple replant disease. The amendments differed in their ability to ameliorate the soil environment; nevertheless, they all promoted the growth of M. hupehensis Rehd. seedlings, and the greatest enhancement of growth was observed in T1. On August 15, 2018, soil urease, sucrase, phosphatase, and catalase activities were 1.67 times, 1.32 times, 1.62 times, and 1.35 times higher in T1 compared with CK, respectively. The soil pH increased, which alleviated soil acidification. T1 also promoted the renewal of the community structure and the diversity of soil microorganisms. The copy numbers of Fusarium solani and Fusarium oxysporum were 71.96 and 70.30% lower in T1 compared with CK, respectively. The seedling height and root length of M. hupehensis Rehd. seedlings increased by 40.97 and 289.69% in T1 compared with CK, respectively. Therefore, soil replanting obstacles can be overcome with the application of quicklime and superphosphate; these soil chemical amendments also improve the soil microbial ecological environment and promote the growth of M. hupehensis Rehd. seedlings.

Amygdalin and Benzoic Acid on the Influences of the Soil Environment and Growth of Malus hupehensis Rehd. Seedlings.

Crop rotation in fruit trees is an effective approach for addressing some of the problems of continuous cropping. To determine whether aged peach orchard soil is suitable for planting apple trees, we studied the effects of two substances abundant in aged peach orchard soil-amygdalin and benzoic acid-on the soil microbial community structure, soil enzyme activity, and the growth of Malus hupehensis Rehd. seedlings. Soils treated with amygdalin (T1), benzoic acid (T2), and a mixed solution of amygdalin and benzoic acid (T3) were used to plant M. hupehensis Rehd. seedlings. Compared with fallow (control) soil, the soil microbial community structure, soil enzyme activities, and root protective enzyme activities, leaf chlorophyll content, and net photosynthetic rate decreased in the three treatments. The biomass and root index of M. hupehensis Rehd. seedlings significantly decreased. Compared with T3, the plant height, ground diameter, fresh weight, dry weight, root length, root surface area, root volume, and root respiration rate of M. hupehensis Rehd. seedlings in T2 in 2015 (2016 in parentheses) decreased by 19.3% (12.6%), 8.7% (7.1%), 21.2% (13.3%), 9.1% (19.6%), 7.9% (25.3%), 40.7% (28.8%), 46.2% (21.1%), and 44.2% (27.5%), respectively. Compared with T3, the same variables in T1 in 2015 (2016 in parentheses) decreased by 34.9% (16.7%), 27.6% (9.8%), 53.6% (19.4%), and 50% (20.5%), 24.1% (31.4%), 55.1% (37.6%), 63.2% (28.2%), and 47.0% (28.7%), respectively. Thus, the inhibitory effect of T3 was the strongest, followed by T2 and T1. In sum, amygdalin and benzoic acid are harmful substances in aged peach orchard soil that inhibit the growth of M. hupehensis Rehd. seedlings.

ITRAQ-based quantitative proteomic analysis of Fusarium moniliforme (Fusarium verticillioides) in response to Phloridzin inducers.

BACKGROUND: Apple replant disease (ARD) has been reported from all major fruit-growing regions of the world, and is often caused by biotic factors (pathogen fungi) and abiotic factors (phenolic compounds). In order to clarify the proteomic differences of Fusarium moniliforme under the action of phloridzin, and to explore the potential mechanism of F. moniliforme as the pathogen of ARD, the role of Fusarium spp. in ARD was further clarified. METHODS: In this paper, the quantitative proteomics method iTRAQ analysis technology was used to analyze the proteomic differences of F. moniliforme before and after phloridzin treatment. The differentially expressed protein was validated by qRT-PCR analysis. RESULTS: A total of 4535 proteins were detected, and 293 proteins were found with more than 1.2 times (P< 0.05) differences. In-depth data analysis revealed that 59 proteins were found with more than 1.5 times (P< 0.05) differences, and most proteins were consistent with the result of qRT-PCR. Differentially expressed proteins were influenced a variety of cellular processes, particularly metabolic processes. Among these metabolic pathways, a total of 8 significantly enriched KEGG pathways were identified with at least 2 affiliated proteins with different abundance in conidia and mycelium. Functional pathway analysis indicated that up-regulated proteins were mainly distributed in amino sugar, nucleotide sugar metabolism, glycolysis/ gluconeogenesis and phagosome pathways. CONCLUSIONS: This study is the first to perform quantitative proteomic investigation by iTRAQ labeling and LC-MS/MS to identify differentially expressed proteins in F. moniliforme under phloridzin conditions. The results confirmed that F. moniliforme presented a unique protein profile that indicated the adaptive mechanisms of this species to phloridzin environments. The results deepened our understanding of the proteome in F. moniliforme in response to phloridzin inducers and provide a basis for further exploration for improving the efficiency of the fungi as biocontrol agents to control ARD.

[Effects of dazomet fumigation on growth, biological characteristics of Malus hupehensis seedlings and soil environment]. / 棉隆熏蒸处理对平邑甜茶幼苗生长和生物学特性及土壤环境的影响.

In this study, we examined the effects of dazomet fumigation with different concentrations (0, 0.1, 0.2, 0.4 g·kg-1) on the microbial characteristics of continuous cropping soil and growth of Malus hupehensis seedling in greenhouse and open-field pot. The results showed that all the treatment of dazomet fumigation could promote the growth of M. hupehensis seedlings in continuous cropping soil, with 0.2 g·kg-1 treatment showing the strongest effect. Compared to the control, plant height, stem diameter, dry weight of M. hupehensis seedlings in 0.2 g·kg-1 dazomet fumigation were increased by 192.9% and 91.8%, 72.8% and 60.1%, 196.8% and 195.0%, 138.5% and 130.7%, respectively in greenhouse and open-field. The root related indices (root length, root area, root volume, root respiration rate) were significantly promoted. The activities of SOD, POD, CAT in roots were increased by 114.6% and 118.5%, 123.5% and 107.6%, 164.6% and 175.6% respectively compared with the control, whereas the content of malondialdehyde was significantly lowered. Soil bacterial content, fungal content, copy number of Fusarium oxysporum gene and soil enzyme activity were significantly decreased with the increasing dazomet concentrations. In conclusion, 0.2 g·kg-1 dazomet fumigation could increase the biomass of M. hupehensis seedlings in continuous cropping, improve soil environment, and effectively alleviate the continuous cropping obstacle. Therefore, 0.2 g ·kg-1 dazomet fumigation could be given priority during the reconstruction of old apple orchards.