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Abstract MYB transcription factors have been linked to anthocyanin synthesis and various color phenotypes in plants. In apple, MYB10 confers a red‐flesh phenotype due to a minisatellite insertion in its R6 promoter, but R6:MYB10 genotypes exhibit various degrees of red pigmentation in the flesh, suggesting the involvement of other genetic factors. Here, it is shown that MdWRKY10, a transcription factor identified via DNA pull‐down trapping, binds to the promoter of MdMYB10 and activates its transcription. MdWRKY10 specifically interacts with the WDR protein MdTTG1 to join the apple MYB‐bHLH‐WDR (MBW) complex, which significantly enhances its transcriptional activation activity. A 163‐bp InDel detected in the promoter region of the alleles of MdWRKY10 in a hybrid population of identical heterozygous genotypes regarding R6 by structural variation analysis, contains a typical W‐box element that MdWRKY10 binds to for transactivation. This leads to increased transcript levels of MdWRKY10 and MdMYB10 and enhanced anthocyanin synthesis in the flesh, largely accounting for the various degrees of flesh red pigmentation in the R6 background. These findings reveal a novel regulatory role of the WRKY‐containing protein complex in the formation of red flesh apple phenotypes and provide broader insights into the molecular mechanism governing anthocyanin synthesis in plants.

Malus sieversii f. niedzwetzkyan, a wild species capable of growing on saline-alkali soil in Xinjiang, is the most promising horticultural crop for improving the saline-alkali wasteland. However, the tolerance of M. niedzwetzkyan to saline-alkali stress and the underlying molecular mechanisms remains largely unknown. Here, we conducted a hydroponic experiment in which M. niedzwetzkyana and M. domestica “Royal Gala” seedlings were subjected to 150 mM saline-alkali stress. Physiological data showed that M. niedzwetzkyana had a strong ROS scavenging ability and ion transport ability, and its saline-alkali resistance was higher than that of M. “Royal Gala”. Saline-alkali stress also promoted the synthesis of anthocyanins in M. niedzwetzkyana. Transcriptome analysis was conducted on the leaves and roots of M. niedzwetzkyana at different time points under saline-alkali stress (0 h, 6 h, and 12 h). Transcriptome analysis revealed that saline stress down-regulated most genes involved in the anthocyanin flavonoid synthesis pathway. Transcription levels of genes involved in antioxidant enzyme activity and ion transport were altered. We identified hub genes related to superoxide dismutase as well as Na+ and K+ transport using weighted gene co-expression network analysis. This study elucidated, for the first time at the molecular level, the saline-alkali tolerance of M. niedzwetzkyana, including the complex changes in pathways that regulate reactive oxygen species homeostasis, ion uptake, and anthocyanoside synthesis under saline-alkali stress conditions. This research provides an important genetic resource for identifying genes involved in responses to saline-alkali stress.

Apple ring rot, which is caused by Botryosphaeria dothidea, severely affects apple production. The mechanisms employed in apple cells against B. dothidea remain unknown. In this research, the pathogen infection mode and the relationship between cell death and disease resistance in ‘Fuji’/B. dothidea interaction pathosystem were investigated. By using transmission electron microscopy (TEM), our research showed that the pathogen infects apple cells both intracellularly and extracellularly. However, compared with that in immature fruit, the incidence of hyphae in the interior of mature apple fruit cells increased dramatically, suggesting that cell wall-mediated penetration resistance could be important in apple resistance against B. dothidea. TEM ultrastructural characterization identified the nuclear morphology of programmed cell death induction in both apple fruit and callus cells under B. dothidea infection. Overexpression of MdVDAC2 (MDP0000271281), which encodes an outer-membrane localized anion channel protein in mitochondria, significantly promoted cell death under B. dothidea infection and simultaneously inhibited pathogen infection, suggesting that cell death represents a disease resistance mechanism in apple against B. dothidea infection. Furthermore, BdCatalase (KAF4307763), a cytochrome P450 family protein BdCYP52A4 (KAF4300696), and subtilisin-domain containing proteins were identified from B. dothidea-secreted proteins, which suggested the potential involvement of active oxygen species and phytoalexins in combating B. dothidea infection and triggering or dampening apple resistance. Collectively, our research suggested that cell wall-mediated penetration resistance, programmed cell death machinery and microbial effector-interrelated signaling were among strategies recruited in apple to combat B. dothidea. The current research laid the foundation for further investigations into resistance mechanisms in apple.

We isolated and identified a bacterium that could produce IAA and degrade phloridzin in the rhizosphere soil of healthy replanted apple (the rootstock is M9T337 and the scion is Yanfu 3), providing a theoretical basis for reducing the obstacles associated with apple replant disease (ARD). Isolates were screened using Salkowski colorimetry and screening medium for phloridzin. The isolate of interest (W6) was identified as Ochrobactrum haematophilum based on morphological analysis, physiological and biochemical tests, and 16S rDNA sequencing. In a laboratory experiment, W6 produced auxin and promoted the growth of Arabidopsis thaliana roots, and its degradation rate of 100 mg · L−1 phloridzin was 62.0%. In a pot experiment, W6 significantly reduced the phenolic acid contents of replanted soil, lowered the abundance of the harmful fungus Fusarium solani, and increased soil enzyme activities, thereby improving the micro-ecological environment of replant soil. W6 increased the root antioxidant enzyme activity and leaf photosynthetic pigment content of replanted Malus hupehensis Rehd. seedlings, effectively alleviating the decrease in net photosynthetic rate, transpiration rate and stomatal conductance caused by ARD. In a field experiment, W6 also promoted the growth of replanted apple (the rootstock is M9T337 and the scion is Yanfu 3) saplings. Therefore, W6 can promote apple growth and degrade phenolic acids, and it can be used as an effective treatment for the reduction of ARD.

Abstract Background Cultivation of resistant rootstocks can effectively prevent apple replant disease (ARD), and grafting tests are an important means of evaluating the compatibility of rootstocks with scions. Methods The apple rootstocks 12–2 (self-named) and Malus hupehensis Rehd. (PYTC) were planted in a replanted 20-year-old apple orchard. The two rootstocks were grafted with scions of 13 apple varieties. Multiple aboveground physiological parameters of the grafted combinations were measured and evaluated to verify the grafting affinity of 12–2 with the scions as compared to Malus hupehensis Rehd. (PYTC). Results The graft survival rate and graft interface healing of 12–2 did not differ significantly from those of PYTC. Mechanical strength tests of the grafted interfaces showed that some mechanical strength indices of Redchief, Jonagold, Starking, Goldspur and Yinv apple varieties were significantly higher when they were grafted onto 12–2 compared to the PYTC control. The height and diameter of shoots and the relative chlorophyll content, photosynthetic and fluorescence parameters, antioxidant enzyme activities and malondialdehyde content of leaves showed that Fuji 2001, Tengmu No.1, RedChief, Gala, USA8, and Shoufu1 grew similarly on the two rootstocks, but Tianhong 2, Lvguang, Jonagold, Starking, Goldspur, Yinv and Luli grew better when grafted onto 12–2 than onto the PYTC control. The rootstock 12-2, therefore, showed good grafting affinity. Conclusion These results provide experimental materials and theoretical guidance for the cultivation of a new grafting compatible rootstock to the 13 studied apple cultivars.

Abstract 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.

Abstract 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.

Abstract Background Elaeagnus angustifolia L. is a deciduous tree in the family Elaeagnaceae. It is widely used to study abiotic stress tolerance in plants and to improve desertification-affected land because of its ability to withstand diverse types of environmental stress, such as drought, salt, cold, and wind. However, no studies have examined the mechanisms underlying the resistance of E. angustifolia to environmental stress and its adaptive evolution. Methods Here, we used PacBio, Hi-C, resequencing, and RNA-seq to construct the genome and transcriptome of E. angustifolia and explore its adaptive evolution. Results The reconstructed genome of E. angustifolia was 526.80 Mb, with a contig N50 of 12.60 Mb and estimated divergence time of 84.24 Mya. Gene family expansion and resequencing analyses showed that the evolution of E. angustifolia was closely related to environmental conditions. After exposure to salt stress, GO pathway analysis showed that new genes identified from the transcriptome were related to ATP-binding, metal ion binding, and nucleic acid binding. Conclusion The genome sequence of E. angustifolia could be used for comparative genomic analyses of Elaeagnaceae family members and could help elucidate the mechanisms underlying the response of E. angustifolia to drought, salt, cold, and wind stress. Generally, these results provide new insights that could be used to improve desertification-affected land.

This study investigated the effects of six crop rotation combinations on the soil quality of old apple orchard and seedling growth of Malus hupehensis Rehd. (apple rootstock) under pot conditions. The inhibitory effects of crops such as Allium fistulosum, Brassica juncea, and Triticum aestivum on four species of Fusarium were observed and compared in six treatments. These were continuous cropping (CK), fumigation with the methyl bromide (FM), rotating A. fistulosum only (R1), rotating A. fistulosum and T. aestivum (R2), rotating A. fistulosum, B. juncea, and T. aestivum (R3), and fallow (FC) in a year. The results showed that the biomass of Malus hupehensis Rehd. seedlings increased significantly. The root length increased and the root architecture was optimized. The respiration rate of the root system was increased by about 1 time after rotation. The treatments of R1, R2, R3, and FC increased bacterial count by 232.17%, 96.04%, 316.21%, and 60.02%, respectively. However, the fungi were reduced in varying degrees and bacteria/fungi ratio was increased by 5–10 times. The enzyme activities, pH, and organic matter were increased, but soil bulk density was decreased. Phenolic acids such as phloridzin was decreased significantly. The copy number of four Fusarium species declined by 85.59%, 74.94%, 69.68%, and 54.41% after rotating three different crops (R3 treatment). The root volatiles of three plants inhibited mycelial growth and spore germination of four Fusarium species.

Abstract Background Light quality significantly affects plant growth and development, photosynthesis, and carbon and nitrogen metabolism. Apple (Malus domestica Borkh.) is a widely cultivated and economically important fruit crop worldwide. However, there are still few studies on the effects of different light qualities on the growth and development of apple seedlings. Results In this study, we explored the effects of blue and red light treatments on the growth and development, photosynthetic characteristics, leaf chloroplast ultrastructure, and carbon and nitrogen metabolism of apple seedlings. Blue light significantly inhibited apple plant growth and leaf extension, but it promoted the development of leaf tissue structures and chloroplasts and positively affected leaf stomatal conductance, the transpiration rate, and photosynthetic efficiency. The red light treatment promoted apple plant growth and root development, but it resulted in loosely organized leaf palisade tissues and low chlorophyll contents. The blue and red light treatments enhanced the accumulation of ammonium nitrogen in apple seedlings. Moreover, the blue light treatment significantly promoted nitrogen metabolism. Additionally, an RNA-seq analysis revealed that both blue light and red light can significantly up-regulate the expression of genes related to carbon and nitrogen metabolism. Blue light can also promote amino acid synthesis and flavonoid metabolism, whereas red light can induce plant hormone signal transduction. The expression of a gene encoding a bHLH transcription factor (MYC2-like) was significantly up-regulated in response to blue light, implying it may be important for blue light-mediated plant development. Conclusions Considered together, blue and red light have important effects on apple growth, carbon and nitrogen metabolism. These findings may be useful for determining the ideal light conditions for apple cultivation to maximize fruit yield and quality.

The study here aimed to investigate the effects of pre-winter ditching and freezing-thawing on soil microbial structure in different soil layers of old apple orchards. A total of 30 samples were obtained from 3 Nov. 2016 to 9 Mar. 2017. The relative abundance, alpha diversity, community structure of fungi, and the relationship between environmental factors and microbial community structure were analyzed, and the greenhouse experiments were used for further verification. Results showed that the number of actinomycete and total bacterial colonies decreased, whereas the number of fungi sustained decreased, resulting in a higher bacteria/fungi ratio. The percentage of Fusarium initially decreased, then later increased by 11.38%, 3.469%, 2.35%, 2.29%, and 3.09%. However, Fusarium levels were still 9% lower on 9 Mar. 2017 that on 3 Nov. 2016. Both the abundance and diversity of the community were higher in the upper soil than in the lower. The main environmental factor contributing to the percentage of Fusarium change was average temperature (AT), although highest temperature (HT) and water content (WC) also had an impact. The Malus hupehensis Rehd. seedlings growing in lower soil were more vigorous than that in upper soil. In sum, pre-winter ditching and freezing-thawing in old apple orchards can reduce the abundance percentage of harmful Fusarium and promote the growth of M. hupehensis Rehd. seedlings.

A pot experiment was performed to investigate the effects of Trichoderma harzianum on the root morphology of Malus hupehensis Rehd. seedlings and their soil environment under replant conditions. The experiment consisted of four treatments: continuously cropped soil (CK1), methyl bromide fumigation (CK2), carrier substrate control (T1), and T. harzianum fertilizer (T2). Plant growth parameters, soil phenolic acid content, abundance of soil microorganisms, and root respiration rate were measured. Compared with CK1, plant height, basal diameter, and fresh weight were 34.58%, 27.55%, and 32.91% greater in T2; 11.35%, 12.10%, and 18.33% greater in T1; and 54.34%, 57.64%, and 45.74% greater in CK2. These metrics were significantly higher in the CK2 treatment than in the other treatments. The second highest values were recorded in the T2 treatment. Differences in root architecture were consistent with differences in biomass. Application of T. harzianum fertilizer was associated with increases of 45.45%, 120.06%, 86.44%, and 268.29% in the activities of the antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX), respectively, and there was little difference between T2 and CK2. The contents of phlorizin and phloretin were 39.39% and 51.70% less in T2, respectively, and 17.85% and 18.14% less in T1, respectively, compared with CK1. Trichoderma harzianum fertilizer increased the abundance of bacteria and actinomycetes while decreasing that of fungi. The gene copy numbers of Fusarium oxysporum and Fusarium moniliforme were 64.30% and 49.35% less, respectively, in the T2 treatment. The fungus population and the gene copy number of Fusarium oxysporum and Fusarium moniliforme was the least in CK2 because of the good sterilization effect. The T. harzianum fertilizer showed satisfactory effects in promoting the root growth of M. hupehensis, increasing the root resistance, decreasing the soil phenolic acid content, and significantly reducing the gene copy number of F. oxysporum and F. moniliforme. In summary, T. harzianum fertilizer is an effective and green alternative for the prevention and control of apple replant disease (ARD).

Apple replant disease (ARD) causes enormous economic loss and threatens the survival of apple industry worldwide. Fusarium solani is one of the pathogens that has been proven to cause ARD. Samples were collected at different time periods to investigate the mechanism of defense responses of apple to F. solani infection by monitoring the biomass, reactive oxygen species (ROS), and antioxidant enzyme activities of the apple rootstock ‘M.9T337’. In addition, the abundance of transcription of four pathogenesis-related (PR) proteins involved in antifungal defense was monitored. The results showed that the apple root system was normal and had small brown areas. However, there is a rapid burst of ROS during the early infection stage, and the activities of antioxidant enzymes and transcription of PRs increased during this period. With the extension in infection time, the infected root tissues displayed dark brown necrosis, and the activities of antioxidant enzymes and abundance of transcription of PRs decreased gradually after reaching their peak. Eventually, the plant biomass decreased, and the plant died. In conclusion, the levels of ROS and activities of antioxidant enzymes played an active role during the early stage of resistance of ‘M.9T337’ apples to infection by F. solani. Infection by F. solani can destroy the ROS scavenging system, causing oxidative damage and inhibiting the growth of apple rootstocks.

Identifying the genetic variation characteristics of phenotypic traits is important for fruit tree breeding. During the long-term evolution of fruit trees, gene recombination and natural mutation have resulted in a high degree of heterozygosity. Apple (Malus × domestica Borkh.) shows strong ecological adaptability and is widely cultivated, and is among the most economically important fruit crops worldwide. However, the high level of heterozygosity and large genome of apple, in combination with its perennial life history and long juvenile phase, complicate investigation of the genetic basis of fruit quality traits. With continuing augmentation in the apple genomic resources available, in recent years important progress has been achieved in research on the genetic variation of fruit quality traits. This review focuses on summarizing recent genetic studies on apple fruit quality traits, including appearance, flavor, nutritional, ripening, and storage qualities. In addition, we discuss the mapping of quantitative trait loci, screening of molecular markers, and mining of major genes associated with fruit quality traits. The overall aim of this review is to provide valuable insights into the mechanisms of genetic variation and molecular breeding of important fruit quality traits in apple.

The relationship between soil texture and the degree of apple replant disease (ARD) was analyzed from the perspective of the microbial community structure and diversity within the rhizosphere soil of Malus hupehensis Rehd. seedlings. Three different textured soils were taken from different apple orchards in Laizhou, Yantai. The soils were divided into two parts, one was kept in replanted conditions, and the other was fumigated with methyl bromide to act as a high standard control. The strength of ARD occurrence was examined by measuring fresh and dry weight suppression (%) of the M. hupehensis seedlings. Differences in the fungal community structure (especially in Fusarium) among the three soil texture types were analyzed using high-throughput sequencing. The results showed that replanted loam clay soil had the highest fungal diversity, followed by sandy loam soil and finally loam soil. The richness of fungi between soil textures, however, was not significantly different. At the genus level, the relative abundance of Fusarium was 1.96%, 0.78%, and 10.89% in replanted sandy loam, replanted loam soil, and replanted loam clay soil, respectively. Moreover, the gene copy number of Fusarium oxysporum, Fusarium solani, and the inhibition rate of fresh weight of M. hupehensis seedlings were the same in the three soil textures. The plant height, photosynthesis (net) (Pn), and stomatal conductance (gS) of the M. hupehensis seedlings were significantly less in the replanted soil compared with the control treatments, with the overall difference being greatest in replanted loam clay soil, followed by replanted sandy loam and then replanted loam soil.

Anthocyanin and proanthocyanidin play important roles in plant secondary metabolism. Although previous studies identified many transcription factors involved in anthocyanin and proanthocyanidin synthesis, the effects of MADS-box transcription factors are unclear in apple. Brassinosteroids (BRs) are steroid hormones that affect plant flavonoid biosynthesis, but the underlying regulatory mechanism is not yet well established. In this study, we identified a MADS-box transcription factor, MdJa2, which contained a highly conserved MADS-box domain and belonged to the STMADS11 subfamily. Additionally, MdJa2 was responsive to BR signal, and the overexpression of MdJa2 inhibited the synthesis of anthocyanin and proanthocyanidin. The silencing of MdJa2 in “Orin” calli promoted anthocyanin and proanthocyanidin accumulations. Moreover, MdJa2 interacted with MdBZR1. MdJa2 was revealed to independently regulate anthocyanin and proanthocyanidin synthesis pathways. The MdJa2–MdBZR1 complex enhanced the binding of MdJa2 to the promoters of downstream target genes. Our research provides new insights into how MADS-box transcription factors in the BR signaling pathway control the accumulations of anthocyanin and proanthocyanidin in red-fleshed apple.

Apple replant disease (ARD) has been reported in all major fruit-growing regions of the world and is often caused by biotic factors (pathogen fungi) and abiotic factors (phenolic compounds). Soil chemical fumigation can kill soil pathogenic fungi; however, the traditionally used fumigant methyl bromide has been banned because of its ozone-depleting effects. There is thus a need to identify greener fumigant candidates. We characterized the effects of different fumigants on the replanted soil environment and the growth characteristics of Malus hupehensis Rehd. seedlings. All five experimental treatments [treatment 1 (T1), metham-sodium; treatment 2 (T2), dazomet; treatment 3 (T3), calcium cyanamide; treatment 4 (T4), 1,3-dichloropropene; and treatment 5 (T5), methyl bromide] promoted significantly the biomass, root growth, and root respiration rate of M. hupehensis seedlings and the ammonium nitrogen (NH4+-N) and nitrate nitrogen (NO3–-N) contents of replanted soil. Metham sodium (T1) and dazomet (T2) had stronger effects compared with 1,3-dichloropropene (T4) and calcium cyanamide (T3). At 172 days after T1, the height, root length, and root respiration rate of Malus hupehensis Rehd. seedlings, and the NH4+-N and NO3–-N contents of replanted soil increased by 91.64%, 97.67%, 69.78%, 81.98%, and 27.44%, respectively, compared with the control. Thus, dazomet and metham sodium were determined to be the optimal fumigants for use in practical applications.

Abstract 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

Melatonin, as a plant growth regulator, is involved in plant stress resistance. We studied the effects of different concentrations (0, 10, 50, 100, 150, and 200 µmol · L−1) of melatonin on the growth and physiological characteristics of strawberry under cadmium (Cd) stress. The results represented that the growth of strawberry seedlings was inhibited under Cd stress, and the seedling biomass, chlorophyll content and the activities of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) decreased. These toxic effects were, however, effectively remediated by exogenous melatonin pretreatment. Melatonin treatment slowed the inhibitory effect of seedling growth and significantly increased seedling biomass, chlorophyll content, antioxidant enzyme activities, and soluble protein levels in leaves and roots compared with the control. Melatonin treatment also reduced the malondialdehyde (MDA) content and the oxidative stress reactions, increased the anthocyanin content, and slowed the senescence rate, thereby reducing the toxic effects of Cd on strawberries. This indicates that foliar application of melatonin can effectively remediate the adverse effects of Cd on strawberry seedlings; the greatest effect was obtained with melatonin concentration of 100 µmol · L−1.

‘Shannongsu’ and ‘Xincixiang’ were two new late-ripening pear cultivars with high quality, that were bred by our team. In order to clear the postharvest storage characteristics, mature pears were collected and stored at room temperature. Several fruit characteristics were evaluated over time, such as firmness, ethylene release rate, content of aroma components, softening-related enzyme activity, and gene expression. Both ‘Shannongsu’ and ‘Xincixiang’ were crisp and juicy stored after 60 d at room temperature, which exhibited excellent storage characteristics. The storability may be comprehensive result of low levels of ethylene production, aroma synthesis, softening-related activities, and gene expression. The research cleared the storage characteristics of ‘Shannongsu’ and ‘Xincixiang’ at room temperature for the first time, which will provide scientific theoretical guidance for fruit production and marketing.

In plants, anthocyanins and proanthocyanidins (PAs) play important roles in plant resistance to abiotic stress. In this study, ozone (O3) treatments caused the up-regulation of Malus crabapple structural genes McANS, McCHI, McANR and McF3H, which promoted anthocyanin and PA accumulation. We identified the WRKY transcription factor (TF) McWRKY71 by screening differentially expressed genes (DEGs) that were highly expressed in response to O3 stress from an RNA sequencing (RNA-seq) analysis. Overexpressing McWRKY71 increased the resistance of ‘Orin’ apple calli to O3 stress and promoted the accumulation of anthocyanins and PAs, which facilitated reactive oxygen species scavenging to further enhance O3 tolerance. Biochemical and molecular analyses showed that McWRKY71 interacted with McMYB12 and directly bound the McANR promoter to participate in the regulation of PA biosynthesis. These findings provide new insights into the WRKY TFs mechanisms that regulate the biosynthesis of secondary metabolites, which respond to O3 stress, in Malus crabapple.

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.

Apple replant disease (ARD) is a common disease in apple producing areas, and more and more evidence shows that soil-borne pathogens are the main factor. However, most of the drugs used to kill microorganisms are not friendly to the environment. Therefore, there is an urgent need to identify a method that can effectively eliminate these harmful microorganisms and to construct a microbial community structure that is conducive to plant growth in the soil. Herein, we use four different application technologies: foliar spraying, foliar soaking, root soaking, and soil soaking, to examine the inhibitory effect of zinc oxide nanoparticles (ZnO-NPs) on ARD. This study found that they all promoted the growth of Malus hupehensis Rehd. seedlings, and the plant height was 1.09 times, 1.15 times, 1.26 times, and 1.36 times higher that of the control, respectively. Soil soaking had the best promotion effect, and the changes in the soil microbial community structure after root soaking were analyzed. After treatment with ZnO-NPs, the abundances of Neocosmospora, Gibberella, and Fusarium were reduced, whereas the abundances of Tausonia, Chaetomium, and Mrakia were increased. The copy numbers of Fusarium solani and Fusarium oxysporum were 55.7 and 68.9% lower in the ZnO-NPs treatment group than those in the control group, respectively. This study found that after ZnO-NPs were applied to the soil, a new microbial community structure that was conducive to plant growth was formed to overcome ARD. In summary, ZnO-NPs, as a green chemical reagent, can overcome ARD, and it can also be applied to other continuous crops.

ABSTRACT: A two-year field experiment was carried out in order to study the occurrence degree and mechanism of apple replant disease (ARD) in the apple orchards with different soil textures. So we can adopt appropriate control measures according to the severity of ARD. Healthy two-year-old seedlings with consistent growth were selected, of which the root stock was T337 and the scion was Yanfu 3. There were significant differences in biomass between methyl bromide fumigation and replanted treatments, and the difference was the largest in clay loam, followed by sandy loam, and loam, which verified ARD in clay loam was most serious, followed by sandy loam and loam. Based on high-throughput sequencing of fungi in soil samples, fungal richness and diversity were the highest in clay loam, followed by sandy loam, and loam. The relative abundance of Fusarium in SX, SL, FX, FL, WX and WL was 7.33%, 19.32%, 2.70%, 4.24%, 10.71% and 23.87%, respectively. Based on Real-time quantitative analysis, there were significant differences in the number of Fusarium oxysporum and Fusarium solani between methyl bromide fumigation and replanted treatments, i.e., clay loam > sandy loam > loam. Fusarium was the main pathogen causing ARD. This shows that ARD is the most serious under replanted clay loam condition. High-throughput sequencing technology was used to prove the difference in Fusarium was one of the important reasons for ARD under different soil textures. This technology provides a new idea for the prevention and control of ARD.

Abstract Background Pear is one of the most important fruit crops worldwide. Anthocyanins and procyanidins (PAs) are important secondary metabolites that affect the appearance and nutritive quality of pear. However, few studies have focused on the molecular mechanism underlying anthocyanin and PA accumulation in pear. Results We conducted metabolome and transcriptome analyses to identify candidate genes involved in anthocyanin and PA accumulation in young fruits of the pear cultivar ‘Clapp Favorite’ (CF) and its red mutation cultivar ‘Red Clapp Favorite’ (RCF). Gene–metabolite correlation analyses revealed a ‘core set’ of 20 genes that were strongly correlated with 10 anthocyanin and seven PA metabolites. Of these, PcGSTF12 was confirmed to be involved in anthocyanin and PA accumulation by complementation of the tt19–7 Arabidopsis mutant. Interestingly, PcGSTF12 was found to be responsible for the accumulation of procyanidin A3, but not petunidin 3, 5-diglucoside, opposite to the function of AtGSTs in Arabidopsis. Transformation with PcGSTF12 greatly promoted or repressed genes involved in anthocyanin and PA biosynthesis, regulation, and transport. Electrophoretic mobility shift and luciferase reporter assays confirmed positive regulation of PcGSTF12 by PcMYB114. Conclusion These findings identify a core set of genes for anthocyanin and PA accumulation in pear. Of these, PcGSTF12, was confirmed to be involved in anthocyanin and PA accumulation. Our results also identified an important anthocyanin and PA regulation node comprising two core genes, PcGSTF12 and PcMYB114. These results provide novel insights into anthocyanin and PA accumulation in pear and represent a valuable data set to guide future functional studies and pear breeding.

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.

The Luochuan area is an important area for apple production in China. With the renewal and transformation of aged apple orchards, the occurrence of apple replant disease (ARD) was inevitable and has seriously affected the sustainable development of apples. Therefore, we randomly selected 14 soil samples from aged apple orchards in the Luochuan area to study the structural changes in the soil fungal community. The results showed that there were significant differences in the diversity of fungal communities between different aged apple orchards. The harmful fungi Gibberella, Fusarium, and Cryptococcus existed in 14 aged apple orchards in the Luochuan area, but their abundances were different in different aged apple orchards. A FUN Guild analysis showed that fungi were mainly present in the aged apple orchards in Luochuan in the saprotroph and pathotroph nutrition modes. Pathogenic fungi were widely present, which increased the risk of disease and seriously affected the growth and development of fruit trees. To sum up, there was a strong correlation between the ages of orchards and the unbalanced microbial community structure. Therefore, pathogenic fungi could be prevented and controlled during the renewal and transformation of aged orchards to reduce the impact of ARD on the apple industry.

Organic acids secreted by plants, such as p-hydroxybenzoic acid, ferulic acid, cinnamic acid, and benzoic acid, can inhibit seed germination and root growth. The effects of root and soil leaching liquor from orchards on the growth of M. hupehensis Rehd. seedlings under sand culture are studied; the seedlings are sampled at 15, 30, 45, and 60 d. Changes in the amount of root exudates are determined using HPLC. Low concentrations of root leaching liquor (A1) and soil leaching liquor (B1) significantly promoted plant growth and chlorophyll synthesis; high concentrations of root leaching liquor (A6) and soil leaching liquor (B4–6) inhibited growth. Low concentrations of soil leaching liquor had no significant effect on the POD, SOD, and CAT activities. A5–6 and B5–6 significantly decreased Fv/Fm and qP values, respectively, and increased NPQ values. All root and soil leaching liquor treatments inhibited the secretion of gallic acid, hydroxybenzoic acid, benzoic acid, and phloridzin, and promoted the secretion of caffeic acid. The root leaching liquor treatments inhibited the secretion of catechin and promoted the secretion of phloretin. The soil leaching liquor treatments promoted the secretion of cinnamic acid. The secretion of other phenolic acids is likely associated with the different concentrations of leaching liquor.

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.

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.

(1) Background: Cultivating resistant rootstocks is an effective way to mitigate apple replant disease (ARD), and we developed superior apple rootstock line 12-2 (self-named), which shows improved ARD resistance. (2) Methods: We used ARD-associated pathogen Fusarium proliferatum MR5 (MR5) to test the fungal infection in the 12-2 line. Seedlings of the 12-2, T337, and M26 rootstock lines were planted in a substrate with potato dextrose broth and MR5 spore solution, and aboveground physiological indicators were measured. (3) Results: MR5 had the greatest effect on the leaf growth of T337 and M26. The incidence rates of infectious symptoms in the T337 and M26 lines were 68 and 100%, respectively. MR5 significantly affected the leaf chlorophyll content, ETR, and NPQ of T337 and M26, as well as Pn and Tr of M26. MR5 tended to reduce the leaf photosynthetic parameters of T337, but the decreases were not significant. The leaf reactive-oxygen-species levels of T337 and M26, the leaf antioxidant-enzyme activities of M26, and the superoxide-dismutase activity of T337 were significantly affected by MR5. MR5 also had a significant effect on the leaf malondialdehyde, proline, and soluble-sugar contents of T337 and M26. None of these aboveground physiological indicators were affected by MR5 in the 12-2 rootstock. (4) Conclusions: The 12-2 rootstock was more resistant to ARD-associated MR5 and could serve as an important test material for resistant-apple-rootstock breeding in China.

Apple replant disease (ARD) is a common soil disease that occurs in apple-growing areas around the world, causing root tip rot and necrosis, plant growth retardation and even plant death. Biofumigation is a promising strategy for controlling ARD due to its advantages of convenient application and being environmentally friendly. Tagetes erecta is an effective biological fumigant, but its effect on ARD is unclear. In the present study, we used Malus hupehensis Rehd. seedlings as the test material to detect the mitigating effects of different forms of T. erecta: air-dried sample (DS), fresh samples (FS) and fresh sample infusion solution (IS) on ARD. The effects of different forms of T. erecta on the growth of apple seedlings, leaf photosynthesis, root antioxidant enzyme, soil enzymatic activity and microbial environment were investigated. Compared with the CK treatments, DS, FS, and IS treatments all significantly increased the biomass of apple seedlings and promoted root growth under replanting conditions. Among them, DS showed the best results. The activity of antioxidant enzyme including superoxide dismutase, peroxidase and catalase were significantly increased in roots grown in soils treated with T. erecta. Moreover, T. erecta treatment also increased the activity of soil urease, phosphatase, sucrase and catalase enzyme, significantly altered the abundance of soil fungal communities and, in particular, reduced the abundance of Fusarium oxysporum, the main causal fungus of ARD. Therefore, our results suggest that biofumigation of different forms of T. erecta enhanced the resistance of ARD by regulating root reactive oxygen levels and improving the soil fungal communities.

Intracellular transport: Identifying how apple pigment is sent for storage The mystery of how pigmenting anthocyanins are transported to intracellular storage is starting to be revealed. Alongside giving fruit their color, anthocyanins confer benefits to plant and human health. Glutathione S-transferases (GSTs) are responsible for transporting anthocyanins to storage vacuoles, yet research is lacking on how GSTs are regulated. Xuesen Chen, from China’s Shandong Agricultural University, and his team analyzed the activity of 23 GST genes in apple, finding one, MdGSTF6, was most highly active in the fruit-coloration stage of apple development. MdGSTF6 expression correlated with fruit anthocyanin levels and also restored anthocyanin levels in transport-suppressed plants, confirming its function. Chen’s team then discovered that MdGSTF6 expression is activated by the protein MdMYB1. The MdMYB1 gene is the major regulatory gene in anthocyanin synthesis, and is now revealed to also influence their transport.

Molecular biology: Making apples even healthier They say that an apple a day keeps the doctor away, but the breeding of new varieties of red-fleshed apples with higher flavonoid contents could make this widely-consumed fruit even healthier. Apples are one of the richest dietary sources of flavonoids—metabolites credited with reducing the risk of various diseases—but during the process of domestication, the genetic diversity and nutritional quality of apples have decreased. In this review, Xue-Sen Chen at Shandong Agricultural University in China and colleagues track the evolution of modern cultivated apples from their ancestor, Malus sieversii, and outline recent developments in our understanding of the molecular pathways underpinning flavonoid synthesis. They also consider how M. sieversii could be used to create new varieties of tasty red-skinned and red-fleshed apples, enriched with beneficial metabolites.

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.

Apple replant disease (ARD), caused largely by soil-borne fungal pathogens, has seriously hindered the development of the apple industry. The use of antagonistic microorganisms has been confirmed as a low-cost and environmentally friendly means of controlling ARD. In the present study, we assessed the effects of Bacillus amyloliquefaciens QSB-6 on the growth of replanted apple saplings and the soil microbial environment under field conditions, thus providing a theoretical basis for the successful use of microbial biocontrol agents. Four treatments were implemented in three apple orchards: untreated replant soil (CK1), methyl bromide fumigation (CK2), blank carrier treatment (T1), and QSB-6 bacterial fertilizer treatment (T2). The plant height, ground diameter, and branch length of apple saplings treated with T2 in three replanted apple orchards were significantly higher than that of the CK1 treatment. Compared with the other treatments, T2 significantly increased the number of soil bacteria, the proportion of actinomycetes, and the activities of soil enzymes. By contrast, compared with the CK1 treatments, the phenolic acid content, the number of fungi, and the abundance of Fusarium oxysporum, Fusarium moniliforme, Fusarium proliferatum, and Fusarium solani in the soil were significantly reduced. PCoA and cluster analysis showed that soil inoculation with strain QSB-6 significantly decreased the Mcintosh and Brillouin index of soil fungi and increased the diversity of soil bacteria in T2 relative to CK1. The soil bacterial community structure in T2 was different from the other treatments, and the soil fungal communities of T2 and CK2 were similar. In summary, QSB-6 bacterial fertilizer shows promise as a potential bio-inoculum for the control of ARD.

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.

Plant defense: Regulating resistance to ring rot A gene with multiple functions in defense against apple ring rot may provide a target for breeding apple varieties resistant to this devastating and enigmatic fungal disease. In China, the world’s greatest producer of apples, apple ring rot poses a severe challenge to growers. A Chinese team led by Shujing Wu and Xuesen Chen, of Shandong Agricultural University, explored the role of a gene, MdSYP121, thought to provide resistance to ring rot. A combination of genetic, genomic, and transgenic experiments showed that MdSYP121 does indeed confer resistance to the disease, and may do this both by regulating hormonal pathways of defense and by stimulating enzymes involved in defensive oxidation/reduction reactions. Overall, their results suggest potential foci for breeding programmes aimed at reducing the susceptibility of apple cultivars to the ring rot 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.

Apple is one of the most important fruit crops. Here, the authors perform deep genome resequencing of 117 diverse accessions and reveal comprehensive models of apple origin, speciation, domestication, and fruit size evolution as well as candidate genes associated with important agronomic traits.

The dwarf M9 (Malus domestica ‘M9’) rootstock is the most widely available Malus rootstock, here we report the complete chloroplast (cp) genome of ‘M9’ rootstock. The size of the complete cp genome was 159,926 bp, with the large-copy (LSC, 88,065 bp) regions, small single-copy (SSC, 19,157 bp) regions, and two inverted repeat regions (IRs, 26,352 bp each). It contained 110 genes, including 78 protein-coding genes, 28 transfer RNA genes (tRNA), and 4 ribosomal RNA genes (rRNA). A phylogenetic tree demonstrated that ‘M9’ rootstock was closely related to M. hupehensis, M. baccata, M. prunifolia, M. micromalus, and M. tschonoskii.

The European wild apple (Malus sylvestris L.) is an important economical fruit crop. In this present study, we characterized the complete chloroplast (cp) genome sequence of Malus sylvestris L. The complete cp genome is 159,926 bp in length with a typical quadripartite structure, containing a large single-copy region (88,064 bp), a small single-copy region (26,353 bp) and a pair of inverted repeat regions (19,157 bp each). A total of 110 unique genes were found in the newly sequenced genome, including 78 protein-coding genes, 28 tRNA genes, and 4 rRNA genes. Of these, 6 protein-coding genes, 7 tRNA genes, and all 4 rRNA genes are duplicated in the inverted regions. A phylogenetic tree was reconstructed using the neighbor-joining method based on the full length of cp genomes within genus Malus. The result showed that M. sylvestris L. was clustered together with the cultivated apple. The complete cp genome could provide valuable information for understanding the phylogenetic relationships within the genus Malus.

Malus baccata var. xiaojinensis belonging to Rosaceae is not only an ornamental tree but also an apple genotype that is highly tolerant to Fe deficiency. In this study, we reported the complete chloroplast (cp) genome of M. baccata var. xiaojinensis using ILLUMINA sequencing. The whole cp genome is 160,067 bp in length, containing a pair of inverted repeats (IRs) of 26,358 bp, a large single copy (LSC) region of 88,157 bp and a small single copy (SSC) region of 19,194 bp. And, the overall GC content of the cp genome was 36.56%. A total of 110 unique genes were found in the cp genome. 17 genes were duplicated in the IRs, including six protein-coding genes, seven tRNA genes, and four rRNA genes. Fourteen genes contained one intron, whereas three genes (rps12, clpP, and ycf3) contained two introns. The phylogenetic analysis demonstrated a close relationship between M. baccata var. xiaojinensis and Malus hupehensis.

AbstractTubby-like proteins (TLPs), which have a highly conserved β-barrel tubby domain, have been found to be associated with some animal-specific characteristics. In the plant kingdom, more than 10 TLP family members were identified in Arabidopsis, rice and maize, and they were found to be involved in responses to stress. The publication of the apple genome makes it feasible to systematically study the TLP family in apple. In this investigation, 9 TLP encoding genes (TLPs for short) were identified. When combined with the TLPs from other plant species, the TLPs were divided into three groups (group A, B and C). Most plant TLP members in group A contained an additional F-box domain at the N-terminus. However, no common domain was identified other than tubby domain either in group B or in group C. An analysis of the tubby domains of MdTLPs identified three types of conserved motifs. Motif 1 and 2, the signature motifs in the confirmed TLPs, were always present in MdTLPs, while motif 3 was absent from group B. Homology modeling indicated that the tubby domain of most MdTLPs had a closed β barrel, as in animal tubby domains. Expression profiling revealed that the MdTLP genes were expressed in multiple organs and were abundant in roots, stems, and leaves but low in flowers. An analysis of cis-acting elements showed that elements related to the stress response were prevalent in the promoter sequences of MdTLPs. Expression profiling by qRT-PCR indicated that almost all MdTLPs were up-regulated at some extent under abiotic stress, exogenous ABA and H2O2 treatments in leaves and roots, though different MdTLP members exhibited differently in leaves and roots. The results and information above may provide a basis for further investigation of TLP function in plants.

In grapevine, the MYB transcription factors play an important role in the flavonoid pathway. Here, a R2R3-MYB transcription factor, VvMYBC2L2, isolated from Vitis vinifera cultivar Yatomi Rose, may be involved in anthocyanin biosynthesis as a transcriptional repressor. VvMYBC2L2 was shown to be a nuclear protein. The gene was shown to be strongly expressed in root, flower and seed tissue, but weakly expressed during the fruit development in grapevine. Overexpressing the VvMYBC2L2 gene in tobacco resulted in a very marked decrease in petal anthocyanin concentration. Expression analysis of flavonoid biosynthesis structural genes revealed that chalcone synthase (CHS), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin reductase (LAR) and UDP glucose flavonoid 3-O-glucosyl transferase (UFGT) were strongly down-regulated in the VvMYBC2L2-overexpressed tobacco. In addition, transcription of the regulatory genes AN1a and AN1b was completely suppressed in transgenic plants. These results suggested that VvMYBC2L2 plays a role as a negative regulator of anthocyanin biosynthesis.

Apple replant disease (ARD) is an important problem in the production of apple. The phenolic acid is one of the causes of ARD. How phenolic acid affects the ARD was not well known. In this study, we analyzed the type, concentration and annual dynamic variation of phenolic acid in soil from three replanted apple orchards using an accelerated solvent extraction system with high performance liquid chromatography (ASE-HPLC). We found that the type and concentration of phenolic acid were significantly differed among different seasons, different sampling positions and different soil layers. Major types of phenolic acid in three replanted apple orchards were phlorizin, benzoic acid and vanillic aldehyde. The concentration of phenolic acid was highest in the soil of the previous tree holes and it was increased from the spring to autumn. Moreover, phenolic acid was primarily distributed in 30-60 cm soil layer in the autumn, while it was most abundant in 0-30 cm soil layer in the spring. Our results suggest that phlorizin, benzoic acid and vanillic aldehyde may be the key phenolic acid that brought about ARD in the replanted apple orchard.