Malus Robusta Research Articles

Determination of adaptive capabilities of various types, forms and varieties of apple trees based on endophytic and epiphytic microbiota indicators

The aim of the research was to identify the most resistant species, forms and varieties of apple trees based on the developmental specifics of the various types of microbiota. The research was conducted in 2014–2023 at the I. V. Michurin Federal Scientific Center, Michurinsk, Tambov region. The year of planting was 2009 with the planning scheme 6×3 m., the rootstock 54–118. The objects of the research are: Malus robusta species; forms 25–8, 64–143, and 54–118; varieties Korichnoe polosatoe, Orlovim, Iyulskoe Chernenko, and Slava Michurinska. The soil type was leached medium-power black soil with a loamy mechanical composition. Pseudomonas syringae bacteria prevailed in resistant plants: 96.6 % in Malus robusta hybrid apple, 89.3 % in Korichnoe polosatoe variety, and 88.7 % in 25–8 form, which is 9.7 %, 2.4 % and 1.8 % respectively, higher than the average value among the test subjects. In 64–143 form and Slava Michurinska variety, the indicators are lower than the average by 5.5 % and 4.2 % respectively. The yield of mixed microbiota in Malus robusta was not observed (0.0 %). Korichnoe polosatoe variety had an indicator below the average value by 0.7 %, in the form 25–8 by 1.3 %, and in the form 64–143 by 11.4 %, which is 4.5 % higher than the average. The magnitude of negative tests ranged from 3.4 % (Malus robusta) to 6.2 % (Iyulskoe Chernenko), which did not exceed the average value. Samples 64–143, 54–118 and Slava Michurinska had above-average values of 1.0 %, 2.0 % and 4.0 %, respectively. According to the complex of indicators of microbiota yield and the percentage of negative tests, Malus robusta, Korichnoe polosatoe variety and 25–8 form had a high potential for environmental resistance. They were recommended for further breeding for resistance to adverse environmental factors.

Comparative transcriptome and hormone analyses of roots in apple among three rootstocks with different rooting abilities

Background Root plays an important role in the growth and development of fruit trees; however, the molecular mechanisms behind the differences among rootstock varie-ties remain unclear. Methods This study examined the effects of different rootstocks on root structure and the endogenous hormone content of 1-year old apple seedlings in combinations of Tianhong 2 (T2)/Malus robusta (HT), T2/G935, and T2/Jizhen 2 (J2). Results The results showed that the T2/HT treatment had greater root length, surface area, volume, average diameter, tips and forks, followed by G935 and J2. In T2/HT leaves and roots, the indole-3-acetic acid (IAA) and gibberellins (GA3) levels were highest, and the abscisic acid (ABA) levels were the lowest. A root transcriptome analysis detected 10,064, 10,511, and 8,719 differentially expressed genes in T2/HT vs. T2/G935, T2/HT vs. T2/J2, and T2/J2 vs. T2/G935, respectively. The analysis of Gene Ontology (GO) terms indicated a significant enrichment in biological processes, cellular components, and molecular functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that plant hormone signaling, MAPK signaling pathway–plant, and plant–pathogen interaction played important roles in differences in the rooting ability of different rootstocks. In addition, some key differential genes were associated with root growth and development and were involved in these metabolic pathways. This study is important for enriching theoretical studies of fruit tree roots.

Genetic diversity and geographic distribution patterns of Malus landraces from 5 cultivated species under climate change in China

China is the largest center of genetic diversity of apple (Malus Mill.), with rich germplasm resources. In the process of evolution and domestication, a large number of apple landraces appeared, and they played a very important role before the introduction of modern cultivated apple varieties to China. However, the habitat of Malus landraces is under serious threat. In this study, 19 pairs of polymorphic simple sequence repeat primers were used to analyze the genetic structure and genetic relationships of Malus landraces and the potential distribution was predicted according to eight environmental variables using MaxEnt modeling. The results showed that the genetic variation of Malus landraces was high (Ho = 0.728 and He = 0.884), and the genetic differentiation was mainly distributed within respective populations and with almost no differentiation among populations. The five population groups were ranked, from highest genetic diversity level to lowest, as follows: Malus domestica subsp. chinensis > Malus prunifolia > Malus robusta > Malus asiatica > Malus micromalus. The 216 germplasms were divided into five groups based on Nei's genetic distance, whereas the Bayesian structure analysis categorized them into two groups. Population structure and principal coordinate analyses showed that Malus domestica subsp. chinensis, Malus asiatica and Malus prunifolia may be derived from similar genetic sources. In the modeling analysis, the cumulative contribution of temperature to the distribution of Malus landraces in suitable growth areas reached 66.1 %, and BIO4 (temperature seasonality) was identified as the major factor influencing the distribution of Malus landraces. Under four future emission scenarios, the potential distribution range of Malus landraces basically unchanged, but the total potential distribution area is projected to grow greatly over time and will tend to shift to higher latitudes.

Multi-omics analysis reveals the biosynthesis of flavonoids during the browning process of Malus sieversii explants.

Malus sieversii is a precious apple germplasm resource. Browning of explants is one of the most important factors limiting the survival rate of plant tissue culture. In order to explore the molecular mechanism of the browning degree of different strains of Malus sieversii, we compared the dynamic changes of Malus sieversii and Malus robusta Rehd. during the whole browning process using a multi-group method. A total of 44 048 differentially expressed genes (DEGs) were identified by transcriptome analysis on the DNBSEQ-T7 sequencing platform. KEGG enrichment analysis showed that the DEGs were significantly enriched in the flavonoid biosynthesis pathway. In addition, metabonomic analysis showed that (-)-epicatechin, astragalin, chrysin, irigenin, isoquercitrin, naringenin, neobavaisoflavone and prunin exhibited different degrees of free radical scavenging ability in the tissue culture browning process, and their accumulation in different varieties led to differences in the browning degree among varieties. Comprehensive transcriptome and metabonomics analysis of the data related to flavonoid biosynthesis showed that PAL, 4CL, F3H, CYP73A, CHS, CHI, ANS, DFR and PGT1 were the key genes for flavonoid accumulation during browning. In addition, WGCNA analysis revealed a strong correlation between the known flavonoid structure genes and the selected transcriptional genes. Protein interaction predictions demonstrated that 19 transcription factors (7 MYBs and 12 bHLHs) and 8 flavonoid structural genes had targeted relationships. The results show that the interspecific differential expression of flavonoid genes is the key influencing factor of the difference in browning degree between Malus sieversii and Malus robusta Rehd., providing a theoretical basis for further study on the regulation of flavonoid biosynthesis.

Bacterial community structure and co-occurrence networks in the rhizosphere and root endosphere of the grafted apple

BackgroundCompared with aerial plant tissues (such as leaf, stem, and flower), root-associated microbiomes play an indisputable role in promoting plant health and productivity. We thus explored the similarities and differences between rhizosphere and root endosphere bacterial community in the grafted apple system.ResultsUsing pot experiments, three microhabitats (bulk soil, rhizosphere and root endosphere) samples were obtained from two-year-old apple trees grafted on the four different rootstocks. We then investigated the bacterial community composition, diversity, and co-occurrence network in three microhabitats using the Illumina sequencing methods. Only 63 amplicon sequence variants (ASVs) out of a total of 24,485 were shared in the rhizosphere and root endosphere of apple grafted on the four different rootstocks (M9T337, Malus hupehensis Rehd., Malus robusta Rehd., and Malus baccata Borkh.). The core microbiome contained 8 phyla and 25 families. From the bulk soil to the rhizosphere to the root endosphere, the members of the phylum and class levels demonstrated a significant enrichment and depletion pattern. Co-occurrence network analysis showed the network complexity of the rhizosphere was higher than the root endosphere. Most of the keystone nodes in both networks were classified as Proteobacteria, Actinobacteriota and Bacteroidetes and were low abundance species.ConclusionThe hierarchical filtration pattern existed not only in the assembly of root endosphere bacteria, but also in the core microbiome. Moreover, most of the core ASVs were high-abundance species, while the keystone ASVs of the network were low-abundance species.

Characterization of Bacillus amyloliquefaciens BA-4 and its biocontrol potential against Fusarium-related apple replant disease.

Apple replant disease (ARD), caused by Fusarium pathogens, is a formidable threat to the renewal of apple varieties in China, necessitating the development of effective and sustainable control strategies. In this study, the bacterial strain BA-4 was isolated from the rhizosphere soil of healthy apple trees in a replanted orchard, demonstrating a broad-spectrum antifungal activity against five crucial apple fungal pathogens. Based on its morphology, physiological and biochemical traits, utilization of carbon sources, and Gram stain, strain BA-4 was tentatively identified as Bacillus amyloliquefaciens. Phylogenetic analysis using 16S rDNA and gyrB genes conclusively identified BA-4 as B. amyloliquefaciens. In-depth investigations into B. amyloliquefaciens BA-4 revealed that the strain possesses the capacity to could secrete cell wall degrading enzymes (protease and cellulase), produce molecules analogous to indole-3-acetic acid (IAA) and siderophores, and solubilize phosphorus and potassium. The diverse attributes observed in B. amyloliquefaciens BA-4 underscore its potential as a versatile microorganism with multifaceted benefits for both plant well-being and soil fertility. The extracellular metabolites produced by BA-4 displayed a robust inhibitory effect on Fusarium hyphal growth and spore germination, inducing irregular swelling, atrophy, and abnormal branching of fungal hyphae. In greenhouse experiments, BA-4 markedly reduced the disease index of Fusarium-related ARD, exhibiting protective and therapeutic efficiencies exceeding 80% and 50%, respectively. Moreover, BA-4 demonstrated plant-promoting abilities on both bean and Malus robusta Rehd. (MR) seedlings, leading to increased plant height and primary root length. Field experiments further validated the biocontrol effectiveness of BA-4, demonstrating its ability to mitigate ARD symptoms in MR seedlings with a notable 33.34% reduction in mortality rate and improved biomass. Additionally, BA-4 demonstrates robust and stable colonization capabilities in apple rhizosphere soil, particularly within the 10-20 cm soil layer, which indicates that it has long-term effectiveness potential in field conditions. Overall, B. amyloliquefaciens BA-4 emerges as a promising biocontrol agent with broad-spectrum antagonistic capabilities, positive effects on plant growth, and strong colonization abilities for the sustainable management of ARD in apple cultivation.

Comparative transcriptomes reveal molecular mechanisms of apple blossoms of different tolerance genotypes to chilling injury.

Apple (Malus domestica, Borkh.) is one of the four largest fruits in the world. Freezing damage during the flowering period of apples is one of the main factors leading to the reduction or even extinction of apple production. Molecular breeding of hardy apples is a good solution to these problems. However, the current screening of cold tolerance genes still needs to be resolved. Therefore, in this article, the transcriptome detection and cold tolerance gene screening during the cold adaptation process of apple were studied in order to obtain potential cold-resistant genes. Herein, two high-quality apple tree species (Malus robusta Rehd and M. domestica) were used for cold adaptation experiments and studied under different low-temperature stress conditions (0, -2 and -4°C). The antioxidant levels of two apple flower tissues were tested, and the transcriptome of the flowers after cold culture was tested by next-generation sequencing technology. Antioxidant test results show that the elimination of peroxides in M. robusta Rehd and the adjustment of the expression of antioxidant enzymes promote the cold resistance of this variety of apples. Functional enrichment found that the expression of enzyme activity, cell wall and cell membrane structure, glucose metabolism/gluconeogenesis, and signal transmission are the main biological processes that affect the differences in the cold resistance characteristics of the two apples. In addition, three potential cold-resistant genes AtERF4, RuBisCO activase 1, and an unknown gene (ID: MD09G1075000) were screened. In this study, three potential cold-resistant genes (AtERF4, RuBisCO activase 1, and an unknown gene [ID: MD09G1075000]) and three cold-repressed differential genes (AtDTX29, XTH1, and TLP) were screened.

Effects of two apple rootstocks on the soil microecology of replanted apple orchard soil

Apple replant disease (ARD) seriously restricts the development of the apple industry. It is of great significance for the sustainable development of the apple industry to alleviate ARD by using rootstock resistance. Taking M9T337 and Malus robusta Rehd. as test materials, the differences in biomass, soil physical and chemical properties, soil enzyme activity, soil microbial metabolic activity and carbon source utilization characteristics, soil microbial quantity, and soil fungal community of two apple rootstocks in replanted soil. The results showed that the biomass, related soil enzyme activities, and AWCD metabolic activity of apple saplings with Malus robusta Rehd. as rootstock were better, the number of soil fungi and the copy number of four Fusarium genes were lower, and more beneficial bacteria were enriched. In conclusion, the apple saplings with Malus robusta Rehd. as rootstock have a better improvement effect on the replanted soil environment and are more suitable as rootstocks to prevent and control ARD than M9T337.

Inoculation with arbuscular mycorrhizal fungi increase calcium uptake in Malus robusta

Utilization of plants-microbe's interactions to remediate calcium11Ca: calcium (Ca) deficiency soils is effective and practical. A greenhouse pot experiment was conducted to evaluate the effect of arbuscular mycorrhizal fungi22AMF: arbuscular mycorrhizal fungi (AMF) inoculation and the synergy of AMF and Ca fertilizer on physiological and transcriptomic responses in apple rootstock (Malus robusta) roots. AMF and Ca fertilizer synergistically served to elongate root length, surface area, average stem diameter and biomass, such extended the surface area in contact with soil nutrients and water. Symbiosis of AMF with apple seedlings made significantly higher levels of Ca in root tissue and promoted Ca translocation from roots to shoot tissue. GO and KEGG pathway analysis disclosed that gene expression varies in several critical gene families, such as auxin response (MdAux/IAAs, MdGH3 and MdSAUR), TCA cycles (MdCS, MdMDH and MdACO), phosphate transporters (MdPHT1;1, MdPHT1;10 and MdPHT1;3) and Ca2+ signal transduction pathways (MdCa2+/ATPase, MdCML, MdTPC1 and MdCDPK), indicating that apple regulates the expression of genes related to auxin synthesis, organic acid secretion, and calcium transporters and channels, thereby increased the growth of apple roots and promoted the secretion of organic acids, resulting in the increase of calcium effectiveness in soil. AMF promoted the accumulation and utilization of Ca in plants by regulating the expression levels of genes related to these pathways. These results contribute to a better understanding of AMF symbiosis mechanism behind the Ca uptake of apple seedlings under exogenous Ca treatment.

Volatile metabolome and floral transcriptome analyses reveal the volatile components of strongly fragrant progeny of Malus × robusta.

Floral fragrance is an important trait that contributes to the ornamental properties and pollination of crabapple. However, research on the physiological and molecular biology of the floral volatile compounds of crabapple is rarely reported. In this study, metabolomic and transcriptomic analyses of the floral volatile compounds of standard Malus robusta flowers (Mr), and progeny with strongly and weakly fragrant flowers (SF and WF, respectively), were conducted. Fifty-six floral volatile compounds were detected in the plant materials, mainly comprising phenylpropane/benzene ring-type compounds, fatty acid derivatives, and terpene compounds. The volatile contents were significantly increased before the early flowering stage (ES), and the contents of SF flowers were twice those of WF and Mr flowers. Odor activity values were determined for known fragrant volatiles and 10-11 key fragrant volatiles were identified at the ES. The predominant fragrant volatiles were methyl benzoate, linalool, leaf acetate, and methyl anthranilate. In the petals, stamens, pistil, and calyx of SF flowers, 26 volatiles were detected at the ES, among which phenylpropane/benzene ring-type compounds were the main components accounting for more than 75% of the total volatile content. Functional analysis of transcriptome data revealed that the phenylpropanoid biosynthesis pathway was significantly enriched in SF flowers. By conducting combined analyses between volatiles and differentially expressed genes, transcripts of six floral scent-related genes were identified and were associated with the contents of the key fragrant volatiles, and other 23 genes were potentially correlated with the key volatile compounds. The results reveal possible mechanisms for the emission of strong fragrance by SF flowers, and provide a foundation for improvement of the floral fragrance and development of new crabapple cultivars.

Receptor-like Kinases (LRR-RLKs) in Response of Plants to Biotic and Abiotic Stresses.

Plants live under different biotic and abiotic stress conditions, and, to cope with the adversity and severity, plants have well-developed resistance mechanisms. The mechanism starts with perception of the stimuli followed by molecular, biochemical, and physiological adaptive measures. The family of LRR-RLKs (leucine-rich repeat receptor-like kinases) is one such group that perceives biotic and abiotic stimuli and also plays important roles in different biological processes of development. This has been mostly studied in the model plant, Arabidopsis thaliana, and to some extent in other plants, such as Solanum lycopersicum, Nicotiana benthamiana, Brassica napus, Oryza sativa, Triticum aestivum, Hordeum vulgare, Brachypodium distachyon, Medicago truncatula, Gossypium barbadense, Phaseolus vulgaris, Solanum tuberosum, and Malus robusta. Most LRR-RLKs tend to form different combinations of LRR-RLKs-complexes (dimer, trimer, and tetramers), and some of them were observed as important receptors in immune responses, cell death, and plant development processes. However, less is known about the function(s) of LRR-RLKs in response to abiotic and biotic stresses. Here, we give recent updates about LRR-RLK receptors, specifically focusing on their involvement in biotic and abiotic stresses in the model plant, A. thaliana. Furthermore, the recent studies on LRR-RLKs that are homologous in other plants is also reviewed in relation to their role in triggering stress response processes against biotic and abiotic stimuli and/or in exploring their additional function(s). Furthermore, we present the interactions and combinations among LRR-RLK receptors that have been confirmed through experiments. Moreover, based on GENEINVESTIGATOR microarray database analysis, we predict some potential LRR-RLK genes involved in certain biotic and abiotic stresses whose function and mechanism may be explored.

Brassinosteroids enhance resistance to manganese toxicity in Malus robusta Rehd. via modulating polyamines profile

Manganese (Mn) toxicity in soil is a widely observed phenomenon, which seriously restricts growth, quality, and yield of various crops and fruits including apples. However, mechanisms underlying the regulation of polyamines (PAs) by brassinosteroids (BRs) to improve tolerance to Mn stress are still unclear. In this study, we investigated the effects of 2,4-epibrassinolide (EBL; a BR) on the expression of genes involved in BR signaling pathway, Mn accumulation, PAs-mediated responses (PA precursor levels, metabolic enzymes, and genes), and growth parameters in Mn-stressed Malus robusta Rehd. EBL application significantly modulated the expressions of genes related to BR signaling (MdBRI, MdBSK, etc.) and reduced Mn accumulation, along with improving the rate of increase in root length and plant height, relative water content, chlorophyll content, maximum photochemical efficiency of PSII (Fv/Fm), and actual photochemical efficiency (ΦPSII) and decreasing electrical conductivity. Furthermore, EBL application significantly reduced putrescine (Put) accumulation and increased spermine (Spm) content and (Spd + Spm)/Put ratio. EBL weakened ornithine (Orn) pathway, decreased ornithine decarboxylase (ODC) activity, and increased biosynthesis of Spm from Put via elevating the PA oxidase (PAO) activity and expression of MdSPDS, MdSPMS, and MdPAO. The trends for free, PS-conjugated, and PIS-bound PAs were similar to that of total PAs, except that no significant change was observed in free Spm, PS-conjugated Spd, and Spm, as well as PIS-bound Spd. This study revealed that BR-regulated PAs help in mitigating Mn toxicity and clarified the mechanisms of regulation of PAs by BRs in apple trees.

The nutrient, hormone, and antioxidant status of scion affects the rootstock activity in apple

In grafted plants, rootstock growth and stress resistance are influenced by scion traits, but the underlying mechanism is unclear in apple. In this study, we analyzed seedlings comprising wild-type (WT) or increased-branching mutant (MB) scions on Malus robusta rootstock on the basis of RNA-seq data, physiological and biochemical indices, and gene expression. Gene functional analysis of RNA-seq data revealed plant–pathogen interaction was the most significantly enriched biological pathway in the axillary buds and roots compared MB with WT. Most genes related to stress responses had up-regulated expression in the MB roots. The leaf water potential, chlorophyll content, and photochemical efficiency were lower in the MB leaves, whereas the water loss rate was higher. The sucrose, glucose, fructose, and sorbitol contents increased significantly in the MB leaves, but decreased in the stems. In the MB roots, there was no change in sucrose and glucose contents, but the fructose and sorbitol contents increased and decreased, respectively. The nitrogen, phosphorus, and potassium contents were higher in the MB roots, while lower nitrogen and phosphorus and higher potassium contents in MB leaves. The abscisic acid, malondialdehyde, and hydrogen peroxide levels increased significantly in the MB seedlings, whereas the antioxidant enzyme activities decreased, especially in the roots and leaves, which was in accordance with the down-regulated expression of the genes related to antioxidant enzyme synthesis. These changes in the MB roots were apparently due to MB scion traits, suggesting that rootstock vitality in grafted apple trees may be affected by the nutrient distribution, hormone level, and antioxidative stress tolerance of the scion.

The Growth-Promoting Mechanism of Brevibacillus laterosporus AMCC100017 on Apple Rootstock Malus robusta

Plant growth-promoting rhizobacteria (PGPR) located in rhizobacteria soil are beneficial to plant growth and development. A PGPR strain AMCC100017 of Brevibacillus laterosporus synthesizes the plant hormone IAA in a tryptophan-dependent manner. In this study, the AMCC100017 strain was used to treat Malus robusta , an excellent natural rootstock for apple production, and assess its ability to promote growth. The fresh weight, dry weight, plant height, and lateral root growth of M. robusta were significantly increased with treatment. The presence of the AMCC100017 strain increased IAA content in M. robusta and promoted root secretion of tryptophan. Colonization of the strain in the roots allowed continuous synthesis of IAA and promoted plant growth. In addition, the photosynthetic efficiency in leaves increased after microbial treatment, and the utilization of nitrogen, zinc, iron, copper and magnesium in leaves was increased, which was conducive to photosynthesis. Interestingly, the activities of CAT and SOD, as well as the contents of ROS in plants colonized by AMCC100017 were increased compared to control plants, but the activities of POD and MDA contents were decreased. AMCC100017 strain affected the antioxidant capacity and stress resistance of plants. AMCC100017 strain increased the content of soluble protein and soluble sugar in plants, improved plant metabolic activity and stress resistance. Therefore, AMCC100017 not only increased IAA content and photosynthetic efficiency to promote M. robusta growth, but also affected plant through multiple metabolic pathways.

Intricate genetic variation networks control the adventitious root growth angle in apple

BackgroundThe root growth angle (RGA) typically determines plant rooting depth, which is significant for plant anchorage and abiotic stress tolerance. Several quantitative trait loci (QTLs) for RGA have been identified in crops. However, the underlying mechanisms of the RGA remain poorly understood, especially in apple rootstocks. The objective of this study was to identify QTLs, validate genetic variation networks, and develop molecular markers for the RGA in apple rootstock.ResultsBulked segregant analysis by sequencing (BSA-seq) identified 25 QTLs for RGA using 1955 hybrids of the apple rootstock cultivars ‘Baleng Crab’ (Malus robusta Rehd., large RGA) and ‘M9’ (M. pumila Mill., small RGA). With RNA sequencing (RNA-seq) and parental resequencing, six major functional genes were identified and constituted two genetic variation networks for the RGA. Two single nucleotide polymorphisms (SNPs) of the MdLAZY1 promoter damaged the binding sites of MdDREB2A and MdHSFB3, while one SNP of MdDREB2A and MdIAA1 affected the interactions of MdDREB2A/MdHSFB3 and MdIAA1/MdLAZY1, respectively. A SNP within the MdNPR5 promoter damaged the interaction between MdNPR5 and MdLBD41, while one SNP of MdLBD41 interrupted the MdLBD41/MdbHLH48 interaction that affected the binding ability of MdLBD41 on the MdNPR5 promoter. Twenty six SNP markers were designed on candidate genes in each QTL interval, and the marker effects varied from 0.22°-26.11°.ConclusionsSix diagnostic markers, SNP592, G122, b13, Z312, S1272, and S1288, were used to identify two intricate genetic variation networks that control the RGA and may provide new insights into the accuracy of the molecular markers. The QTLs and SNP markers can potentially be used to select deep-rooted apple rootstocks.

Genomics-assisted prediction of salt and alkali tolerances and functional marker development in apple rootstocks

BackgroundSaline, alkaline, and saline-alkaline stress severely affect plant growth and development. The tolerance of plants to these stressors has long been important breeding objectives, especially for woody perennials like apple. The aims of this study were to identify quantitative trait loci (QTLs) and to develop genomics-assisted prediction models for salt, alkali, and salt-alkali tolerance in apple rootstock.ResultsA total of 3258 hybrids derived from the apple rootstock cultivars ‘Baleng Crab’ (Malus robusta Rehd., tolerant) × ‘M9’ (M. pumila Mill., sensitive) were used to identify 17, 13, and two QTLs for injury indices of salt, alkali, and salt–alkali stress via bulked segregant analysis. The genotype effects of single nucleotide polymorphism (SNP) markers designed on candidate genes in each QTL interval were estimated. The genomic predicted value of an individual hybrid was calculated by adding the sum of all marker genotype effects to the mean phenotype value of the population. The prediction accuracy was 0.6569, 0.6695, and 0.5834 for injury indices of salt, alkali, and salt–alkali stress, respectively. SNP182G on MdRGLG3, which changes a leucine to an arginine at the vWFA-domain, conferred tolerance to salt, alkali, and salt-alkali stress. SNP761A on MdKCAB, affecting the Kv_beta domain that cooperated with the linked allelic variation SNP11, contributed to salt, alkali, and salt–alkali tolerance in apple rootstock.ConclusionsThe genomics-assisted prediction models can potentially be used in breeding saline, alkaline, and saline-alkaline tolerant apple rootstocks. The QTLs and the functional markers may provide insight for future studies into the genetic variation of plant abiotic stress tolerance.

Effects of Exogenous Manganese (Mn) on Mineral Elements, Polyamines and Antioxidants in Apple Rootstock Malus robusta Rehd.

Manganese (Mn) is one of the essential microelements in all organisms. However, high level of Mn is deleterious to plants. In this study, the effects of exogenous manganese application on mineral element, polyamine (PA) and antioxidant accumulation, as well as polyamine metabolic and antioxidant enzyme activities, were investigated in Malus robusta Rehd., a widely grown apple rootstock. High level of Mn treatments decreased endogenous Mg, Na, K and Ca contents, but increased Zn content, in a Mn-concentration-dependent manner. Polyamine metabolic assays revealed that, except the content of perchloric acid insoluble bound (PIS-bound) spermine, which increased significantly, the contents of putrescine (Put), spermidine (Spd) and spermine (Spm) all decreased progressively, accompanied with the decreased activities of arginine decarboxylase (ADC, EC 4.1.1.19) and ornithine decarboxylase (ODC, EC 4.1.1.17), and the increased activities of diamine oxidase (DAO, EC 1.4.3.6) and polyamine oxidase (PAO, EC 1.5.3.3). Further antioxidant capacity analyses demonstrated that contents of anthocyanin, non-protein thiols (NPT) and soluble sugar, and the activities of guaiacol peroxidase (POD, EC 1.11.1.7), catalase (CAT, EC 1.11.1.6) and superoxide dismutase (SOD, EC 1.15.1.1), also increased upon different concentrations of Mn treatments. Our results suggest that endogenous ion homeostasis is affected by high level of Mn application, and polyamine and antioxidant metabolism is involved in the responses of M. robusta Rehd. plants to high level of Mn stress.

Apple rootstocks with different phosphorus efficiency exhibit alterations in rhizosphere bacterial structure.

The purpose of this study was to select phosphorus-efficient apple rootstocks under phosphorus deficiency and to reveal the effects of different apple rootstocks on the rhizosphere bacterial community. We used 83 hybrid lines of Malus robusta Rehd.×Malling 9 (M.9) to investigate their physiological traits and the phosphorus deficiency phenotypes of leaves in response to phosphorus deficiency (0·1mmoll-1 PO4 3- ). All the plants were cultivated in pots in the greenhouse and watered using drip irrigation. In accordance with the results of investigation, we selected the phosphorus-efficient hybrid lines (PE) and the phosphorus-inefficient hybrid lines (PI) to research their root morphology and root hairs (RH). In addition, we used Illumina MiSeq sequencing to determine the bacterial community of the rhizosphere from different rootstocks. The results showed that the PE plants had better growth characteristics and stronger root plasticity than that of the PI plants, and phosphorus deficiency can stimulate the RH growth of PE plants. There was no significant difference in the rhizosphere bacterial diversity, but we found that the bacterial community structure was significantly different at the genus levels; in addition, 89 genera were found to have significant differences between PE and PI plants, especially Bacillus. The PE rhizosphere had more abundant Bacillus compared to the PI. High positive Pearson correlations with the phosphorus concentration in the plantlets of apple rootstocks were detected for the bacterial genera Bacillus (r: 0·776). The phosphorus-efficient apple rootstocks adapted to phosphorus deficiency by shaping the root morphology. Notably, different apple rootstocks showed alteration of the microbes in rhizosphere. This study can provide the materials for exploring the mechanism of apple rootstock phosphorus absorption. In accordance with the different bacterial community compositions, we can develop the inoculants to promote nutrient uptake.

Tolerance of two apple rootstocks to short-term salt stress: focus on chlorophyll degradation, photosynthesis, hormone and leaf ultrastructures

To elucidate the salt tolerance of Malus halliana Koehne and Malus robusta Rehd., changes of photosynthetic parameters, hormone content and chlorophyll degradation enzymes of them were compared after treated with different concentration of NaCl. Salt stresses were simulated using 50, 100, 200 mM of NaCl solution, and 1/2 Hoagland nutrient solution was used instead of NaCl solution as control (CK). Except for the indole acetic acid (IAA) content, the changes of Chlorophyll (Chl) content, net photosynthetic rate (PN), stomatal conductance (gs), IAA/ABA ratio, intercellular CO2 concentration (Ci), transpiration rate, abscisic acid content (ABA), Pheophytinase (PPH) and Pheophorbide a monooxygenase (PaO) in Malus halliana were lower than those in Malus robusta under 100 and 200 mM NaCl condition. The values of Chlase, PPH and Pao in Malus robusta were significantly higher than that in Malus halliana. According to correlation analysis, the Chl was extremely positively correlated with Chla, Chlb, IAA/ABA, PN and gs, significantly negative correlation with Chla/b, ABA and PaO, and negative correlation with Ci, Chlase and PPH. The chloroplast ultrastructure of Malus robusta was greatly damaged resulting in chloroplasts disintegration and that of Malus halliana was maintained to be completed at 100 and 200 mM NaCl. In certain range of salt concentrations, Malus halliana could be better to adjust the levels of Chlorophyll content, Chlase, PPH, PaO and the number of starch grain and osmiophilic granules to relieve the damage of photosynthetic system due to salt stress. In a word, the Malus halliana could be better adapted to high salt concentration than Malus robusta.

Apple rootstocks of different nitrogen tolerance affect the rhizosphere bacterial community composition.

To select apple rootstocks that are tolerant to low nitrogen and reveal the relationship between the rhizosphere bacterial communities and the low nitrogen tolerance of the apple rootstock. In total, 235 lines of hybrids of Malus robusta Rehd.×M.9 with low nitrogen stress were cultivated in pots in a greenhouse equipped with a drip irrigation system, and growth characteristics, photosynthesis traits and mineral elements were monitored. The bacterial community structure of the rhizosphere from different rootstocks was determined via Illumina MiSeq sequencing. This study selected three low nitrogen-tolerant (NT) lines that had higher nitrogen concentration, and higher photosynthesis rate than the three low nitrogen-sensitive (NS) lines. The bacterial community structure significantly differed (P≤0·001) among therootstocks. The bacterial phyla Proteobacteria and Actinobacteria were the dominant groups in the rhizosphere and presented higher abundance in the NT rhizosphere. The N concentration in the apple rootstocks exhibited highly positive Pearson correlations with the bacterial genera Sphingomonas, Pseudoxanthomonas, Bacillus and Acinetobacter, and negative correlations with the bacterial genera Pseudarthrobacter and Bradyrhizobium. This study showed that investigated rootstocks achieved increased nitrogen concentration by enhancing their photosynthetic production capacity and shaping their rhizobacteria community structure. The findings provide a basis for studying the mechanisms of resistance to low nitrogen stress in apple rootstocks. Based on these beneficial bacteria, microbial inoculants can be developed for use in sustainable agricultural and horticultural production.