Nematode–Microbe Complexes in Soils Replanted with Apple

Apple replant disease is a severe problem in orchards and tree nurseries. Evidence for the involvement of a nematode–microbe disease complex was reported. To search for this complex, plots with a history of apple replanting, and control plots cultivated for the first time with apple were sampled in two fields in two years. Shoot weight drastically decreased with each replanting. Amplicon sequencing of the nematode community and co-extracted fungal and bacterial communities revealed significant differences between replanted and control plots. Free-living nematodes of the genera Aphelenchus and Cephalenchus and an unidentified Dorylaimida were associated with replanted plots, as indicated by linear discriminant analysis effect size. Among the co-extracted fungi and bacteria, Mortierella and Methylotenera were most indicative of replanting. Some genera, mostly Rhabditis, Streptomyces and a fungus belonging to the Chaetomiaceae indicated healthy control plots. Isolating and investigating the putative disease complexes will help to understand and alleviate stress-induced root damage of apple in replanted soil.

Control of Apple Replant Disease Using Mixed Cropping with Brassica juncea or Allium fistulosum

Evidence indicates that Allium and Brassica species which release bioactive compounds are widely used in bio-fumigation to suppress soil-borne diseases. However, the active molecules of such plant residues are easily volatilized. In this study, we conducted mixed cropping of the apple tree with Allium fistulosum or Brassica juncea; the results demonstrated that such mixed cropping significantly improved the growth of the grafted apple seedlings and alleviated apple replant disease (ARD) for two years. The terminal-restriction fragment length polymorphism profile results showed that the soil fungal community demonstrated distinct variation and diversity in terms of composition. A. fistulosum and B. juncea significantly improved the Margalef, Pielou, and Shannon indices. In addition, the analyses of clone libraries showed that A. fistulosum and B. juncea promoted the proliferation of antagonistic fungi such as Mortierella, Trichoderma, and Penicillium, and inhibited the proliferation of pathogens such as Fusarium. Fusarium. Proliferatum (F. proliferatum) was abundant in replanted soil and proved to be an aggressive pathogen of apple seedlings. Our findings thus indicate that apple tree mixed cropping with A. fistulosum and B. juncea was an effective long-term method for modifying the resident fungal community and alleviating ARD.

Nematode-Microbe Complexes in Soils Replanted with Apple

Apple replant disease is a severe problem in orchards and tree nurseries. Evidence for the involvement of a nematode-microbe disease complex was reported. To search for this complex, plots with a history of apple replanting, and control plots cultivated for the first time with apple were sampled in two fields in two years. Shoot weight drastically decreased with each replanting. Nematodes were extracted from soil samples by floatation-centrifugation, washed on a 20 µm-sieve, and used for DNA extraction. Nematode communities and co-extracted fungi and bacteria were analyzed by high-throughput sequencing of amplified ribosomal fragments. The nematode community and co-extracted fungal and bacterial communities significantly differed between replanted and control plots. Free-living nematodes of the genera Aphelenchus, Cephalenchus, and an unidentified Dorylaimida were associated with replanted plots, as indicated by linear discriminant analysis effect size. Among the co-extracted fungi and bacteria, Mortierella was most indicative of replanting. Some genera, mostly Rhabditis, indicated healthy control plots. Isolating and investigating the putative disease complexes will help to understand and alleviate stress-induced root damage of apple in replanted soil.

Detection of Root Physiological Parameters and Potassium and Calcium Currents in the Rhizoplane of the Apple Rootstock Superior Line 12-2 With Improved Apple Replant Disease Resistance

The cultivation of resistant rootstocks is one of the more effective ways to mitigate apple replant disease (ARD). We performed an ion current test, a pot experiment, and a pathogen infection test on the apple rootstocks 12-2 (self-named), T337, and M26. The ion current test showed that exposure to ARD soil extract for 30 min had a significant effect on K+ ion currents at the meristem, elongation, and mature zones of the M26 rhizoplane and on Ca2+ currents in the meristem and elongation zones. ARD also had a significant effect on Ca2+ currents in the meristem, elongation, and mature zones of the T337 rhizoplane. Exposure to ARD soil extract for 5 min had a significant effect on K+ currents in the meristem, elongation, and mature zones of 12-2 and on the Ca2+ currents in the elongation and mature zones. Compared to a 5-min exposure, a 30-min exposure to ARD extract had a less pronounced effect on K+ and Ca2+ currents in the 12-2 rhizoplane. The pot experiment showed that ARD soil had no significant effect on any root architectural or physiological parameters of 12-2. By contrast, ARD soil significantly reduced some root growth indices and the dry and fresh weights of T337 and M26 compared with controls on sterilized soil. ARD also had a significant effect on root metabolic activity, root antioxidant enzyme activity (except superoxide dismutase for T337), and malondialdehyde content of T337 and M26. Pathogen infection tests showed that Fusarium proliferatum MR5 significantly affected the root structure and reduced the root metabolic activity of T337 and M26. It also reduced their root antioxidant enzyme activities (except catalase for T337) and significantly increased the root malondialdehyde content, reactive oxygen levels, and proline and soluble sugar contents. By contrast, MR5 had no such effects on 12-2. Based on these results, 12-2 has the potential to serve as an important ARD-resistant rootstock.

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.

The Effects of Bioinoculants Based on Mycorrhizal and Trichoderma spp. Fungi in an Apple Tree Nursery under Replantation Conditions

Both mycorrhizal and Trichoderma spp. fungi are known for antagonistic effects against certain biological pathogens causing apple replant disease (ARD). The aim of this study was to assess the effectiveness of the bioinoculants based on endomycorrhizal and Trichoderma spp. fungi on the biological properties of soil as well as the parameters of the apple tree growths in a fruit tree nursery under replantation conditions. A two-year experiment was conducted on Jonagold apple trees grafted on to M.9 rootstock in western Poland. The trees were planted in the replant soil—from areas used for the production of apple trees, and in the crop rotation soil, that had not been used for nursery purposes before. A mycorrhizal inoculum and preparations containing Trichoderma spp. fungi were applied to the replant soil. Biological properties of the soil and the growth of the aerial and underground parts of the apple trees were assessed. The enzymatic (dehydrogenases and protease) and respiratory activity of the replant soil was significantly lower than that of the crop rotation soil. The apple trees grew worse when exposed to the ARD conditions. The effectiveness of applied bioinoculants in mitigating the effects of replantation in the nursery were shown. Both the treatment mycorrhization and the application of bioinoculants containing Trichoderma spp. increased the respiratory and enzymatic activity of the replant soil. The growth of the root system and the aerial parts of the trees (including leaves) was much better after the combined use of both types of fungi than in the replant soil that had not received the fungal treatment.

Biocontrol Potential of The Phloridin-Degrading Bacillus Licheniformis XNRB-3 Against Apple Replant Disease

Background: Apple replant disease (ARD) is a common occurrence in many major apple-growing areas worldwide, seriously hindering the development of the apple industry. To avoid the shortcomings of chemical fungicides currently used to control ARD, it is necessary to find sustainable and effective control methods. Here, an endophytic phloridin-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 its how to alleviates the pathogen pressure via changes in soil microbiomes were studied.Results: 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 reached more than 60%. The fermentation broth can also protect the roots of the plants from Fusarium 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 and indirect interspecific competition for nutrients. The antifungal organic compounds in the fermentation metabolites were identified using 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 can promote the root elongation and lateral root development of Arabidopsis plants. The potential of strain XNRB-3 to control ARD was later validated using microbial fertilizer inoculation in pot and field experiment. The addition of strain XNRB-3 significantly promoted the growth of plants, and the activity of enzymes related to disease resistance (SOD, POD, and CAT) was also significantly enhanced. It also reduced the abundance 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 soil enzyme activity. Conclusions: The incorporation of strain XNRB-3 in the soil alleviated the damage of soil-borne pathogens to plants by reducing the relative abundance of pathogenic fungi and the content of phenolic acids, and inducing disease resistance of plants. Taken together, B. licheniformis XNRB-3 could be developed into a promising biocontrol and plant-growth-promoting agent. This provides a new management strategy to control ARD.