Wild Apple Research Articles

Identification and Mining of Functional Components of Polyphenols in Fruits of Malus Germplasm Resources Based on Multivariate Analysis.

Polyphenols are important functional components that have anti-cancer and anti-inflammatory effects. Apple fruit is rich in polyphenols and is one of the dietary sources of polyphenols. The polyphenol components and contents of the peel and pulp of 74 Malus sieversii (Led.) Roem. and 26 Chinese Malus germplasm resources were determined using ultra-high-phase chromatography (UPLC) and liquid chromatography-mass spectrometry (LC-MS). The results showed that 34 components were detected in the peel and 30 in the flesh, and that the polyphenol components and contents of the different germplasm resources were significantly different; the polyphenol content of Malus sieversii (Led.) Roem. was significantly higher than that of the other local varieties, and the polyphenol content in the peel was also higher than that in the flesh. Rutin, quercetin 3-O-arabopyranoside, kaempferol 3-O-rutinoside, and peonidin 3-O-galactoside were detected only in the peel. The total polyphenol content in the peel ranged from 949.76 to 5840.06 mg/kg, and the polyphenol content in the pulp ranged from 367.31 to 5123.10 mg/kg. The cluster analysis of polyphenol components and contents in peel and pulp showed that 100 Malus germplasm resources could be grouped into four categories. Principal component analysis of 34 kinds and 30 kinds of polyphenols in peel and pulp of 100 resources was performed. If the eigenvalue is greater than 1, eight and seven principal components are extracted, respectively. Five Malus resources with high polyphenol content in the peel and pulp were selected: 'XY-77' (peel: 5840.06 mg/kg, pulp: 5123.10 mg/kg; 'LF-09' (peel: 4692.63 mg/kg, pulp: 3729.79 mg/kg); '2012-5' (peel: 4377.61 mg/kg, pulp: 3847.54 mg/kg); '29028' (peel: 5088.05 mg/kg, pulp: 3994.61 mg/kg); and '11-01' (peel: 5154.45 mg/kg, pulp: 3616.15 mg/kg). These results provide us with information regarding the polyphenol composition and content of the wild apple resources and local cultivars. The high polyphenol content resources obtained by screening can be used as raw materials for the extraction of polyphenol components and functional fruit juice processing and can also be used as parents for functional fruit creation and variety breeding.

Dynamic Changes in Polyphenols in Fruit Development of Red Flesh Apple ‘Hongxun 2’

In this study, fruits of the red flesh Malus plant ‘Hongxun 2’ (Malus neidzwetzkyana (Dieck) Langenf.) and green flesh Malus plant ‘Xinye 13-11’ (Malus sieversii (Led.) Roem.) were used as experimental materials. Both of them came from Xinjiang, China, and Malus neidzwetzkyana (Dieck) Langenf. is believed to be a variant of Malus sieversii (Led.) Roem. The components and contents of polyphenols in the peel and pulp of the two kinds of fruit during the development period were detected, and the dynamic changes and differences in the polyphenols between the two kinds of fruit were discussed. The results showed that the total polyphenol content of ‘Xinye 13-11’ was higher in the peel and pulp than that of ‘Hongxun 2’, and the content of peel was higher than that of pulp in the two kinds of fruit. An analysis of five types of polyphenols showed that anthocyanins were only contained in the peel and pulp of ‘Hongxun-2’, and the peel had a higher content than the pulp. Cyanidin 3-O-galactoside was the main anthocyanin component. Four other types of substances, except hydroxycinnamics, were higher in ‘Hongxun-2’ than ‘Xinye 13-11’, while the contents of other substances in ‘Xinye 13-11’ were higher than those of ‘Hongxun 2’. The accumulation of major polyphenol components in the peel and flesh of ‘Hongxun 2’ and ‘Xinye 13-11’ apples was significant in the period before and after 65 days after flowering, and the contents of procyanidin B1 and procyanidin C1 were the highest in this period. In addition to the difference in anthocyanin content between ‘Hongxun 2’ and ‘Xinye 13-11’, the chlorogenic acid content in the peel and pulp of ‘Hongxun 2’ was significantly higher than that of ‘Xinye 13-11’, and the contents of other components were lower than those of ‘Xinye 13-11’. Moreover, based on the components and contents of polyphenol components, this paper supports the viewpoint that Malus neidzwetzkyana (Dieck) Langenf is a separate species to Malus sieversii (Led.) Roem.

Two new species of Cytospora (Diaporthales, Cytosporaceae) causing canker disease of Malusdomestica and M.sieversii in Xinjiang, China.

Apple tree canker is a serious disease caused by species of Cytospora. Xinjiang Uygur Autonomous Region is one of the most important apple-producing areas in China. However, losses due to apple Cytospora canker have seriously damaged the apple industry and affected the economic development of the apple growers in this region. In this study, we used morphological characteristics combined with multilocus phylogenetic analyses of the ITS, act, rpb2, tef1 and tub2 loci to identify isolates from apple (Malusdomestica) and wild apple (M.sieversii). As a result, C.hippophaopsis sp. nov. from M.sieversii and C.shawanensis sp. nov. from M.domestica were discovered and proposed herein. Pathogenicity tests were further conducted on 13 varieties of apple and wild apple, which confirmed C.hippophaopsis and C.shawanensis as canker pathogens. Meanwhile, C.hippophaopsis is generally more aggressive than C.shawanensis on the tested varieties of apple and wild apple.

Potential Ecological Distribution of the Beetle Agrilus mali Matsumura (Coleoptera: Buprestidae) in China under Three Climate Change Scenarios, with Consequences for Commercial and Wild Apple Forests.

The apple jewel beetle (AJB), Agrilus mali Matsumura (Coleoptera: Buprestidae), is a dangerous pest of commercial apple orchards across China, the largest apple production country in the world, and has recently become invasive in the Xinjiang Uygur Autonomous Region (XUAR) of northwestern China, where wild apple forests also occur. This pest poses a serious threat to apple production and wild apple forests throughout the world. Global warming is expected to change the geographical distribution of A. mali in China, but the extent of this is unknown. Based on empirical data from 1951 to 2000, a MaxEnt model was used to forecast the ecological distribution of A. mali under three different climate scenarios projected in the fifth report of the Intergovernmental Panel on Climate Change. The results showed that the most important variables were the maximum temperature of November, precipitation in January, and minimum temperatures in April. Under all climate scenarios, the forecasted suitable regions for A. mali in China will expand northward in the 2050s and 2070s. The forecasted highly suitable regions will be 1.11-1.34 times larger than they are currently, and their central distributions will be 61.57-167.59 km further north. These findings suggest that the range and damage caused by A. mali in China will increase dramatically in the future. Monitoring and management measures should be implemented urgently to protect both the commercial apple industry and wild apple resources.

Identifying Fire Blight-Resistant Malus sieversii Rootstocks Grafted with Cultivar ‘Aport’ Using Monitoring Data

In the present study, the most valuable cultivar ‘Aport krovavo-krasnyy’ was grafted onto M. sieversii genotypes harvested from 11 populations in Dzungarian Alatau and Ile Alatau to identify ones resistant to Erwinia amylovora. The wild apple populations included in the present research have not been previously explored. Seedling population 10, developed using rootstocks from a M. sieversii population growing in Turgen, demonstrated the highest resistance to Erwinia amylovora, showing no fire blight symptoms and no positive PCR results for E. amylovora during the eight years of monitoring in the Talgar field (Kazakhstan) from 2015 to 2022. The population from Steep Tract (seedling population 1) was also valuable for breeding and reduced the pathogen distribution to below 30%. Genotypes from a genetic reserve (seedling population 5) were the most susceptible among the researched populations, with a disease distribution level of 24–95%. In seedling population 5, trees affected at least twice by the pathogen exhibited wilting, shepherd’s crook formation, leaf necrosis, and occasional exudate droplets, while trees in other combinations primarily showed shoot wilting and leaf death. Fire blight disease also developed more rapidly within the plant in seedling population 5; by 2020, one tree nearly died after only two infections.

Morphological and biochemical characteristics of wild red-fleshed apples (Malus sieversii f. niedzwetzkyana) in the North and Northeast of Iran

BackgroundRed-flesh apples (Malus sieversii f. niedzwetzkyana) have attracted attention from consumers and researchers due to their pleasant appearance and taste. These exotic apples are rich sources of nutrients and health-promoting polyphenols and phenolics. This study aimed to investigate morphological (40 quantitative and 13 qualitative traits) and biochemical (5 traits) characteristics of four socioeconomically important red-fleshed apple populations in North and Northeast region of Iran, which are understudied and under serious extinction risk.ResultsThe examined characters exhibited wide and statistically significant variations, especially in leaf color (68.86%) and the number of seeds per fruit (61.61%), and more dramatically in the total flavonoids (146.64%) and total phenolics contents (105.81%). There were also strong variations in fresh fruit weight and fruit length, diameter, and flesh thickness. Red, with 34 accessions, was the dominant ripe fruit skin color. All biochemical traits also showed high variations, particularly in total flavonoid content. Red-fleshed Gavramak and Kalateh Khij apples contained the highest biochemical and morphological values, respectively. Principal component analysis (PCA) revealed that the first five principal components together accounted for more than 60.83% variation of the total observed variations. Moreover, the cluster dendrogram analysis based on Ward’s method indicated three different clusters based on the characters measured, indicating high variation among the accessions.Conclusionred-flesh apples can be considered suitable sources of natural antioxidants with great potential as healthy foods and nutraceutical applications. Based on the commercial characters, Red-fleshed Gavramak and Kalateh Khij apples showed the highest fruit quality with proper size and thus can be suggested as superior for cultivation or use in breeding programs due to having higher quality fruits.

Beneath the apple trees - Exploring soil microbial properties under Malus domestica concerning various land management practices

The presented study evaluated the impact of six land management practices on soil bacterial and fungal communities under apple trees: green field belts, forests, gardens with trees and ornamental plants, gardens with farm animals, and uncultivated and cultivated orchards. The study explored soil microbial properties including the enzymatic activity of dehydrogenases (tested by colorimetric reaction with 2,3,5-triphenyl-tetrazolium chloride), metabolic profile (using Biolog™ ECO Plates), metataxonomy structure (Next Generation Sequencing using Illumina®), followed by physicochemical properties (pH, N, P, K, microelements concentrations, organic matter, and Corg content). The hypothesis was that different land management practices would influence soil microbial properties, with cultivated orchards expected to show significantly lower dehydrogenases activity, and higher substrate-based respiratory than biomass response, within substrate stress occurrence, when testing metabolic profiles, but also different composition and lower relative abundances of specific microbial taxa and different biomarker genera, compared to other treatments. We aimed to identify practices promoting diverse microbial substrate-based metabolic and taxa diversity responses, with a focus on Bacillus and Trichoderma abundances, which are potential biological agents against fungal pathogens. As expected the presented research revealed significant statistical variations in microbial communities among different land management practices in soil beneath apple trees. It was accordingly noted that cultivated orchards, but also green belts, clearly exhibited reduced microbial activity (3.59 and 4.76 TPF kg−1 d−1, respectively) compared to gardens and uncultivated orchards (12.08 and 9.89 TPF kg−1 d−1). Cultivated orchards notably showed higher respiration levels and substrate stress compared, especially to forests and other land management practices represented by a clear separation of observed according to Sneath's criteria in cluster analysis. Different land management practices induce unique stress responses in microbial communities: forests struggled with B-Methyl-d-Glucoside, gardens with Serine and Putrescine, cultivated orchards with d-Glucosaminic Acid and Cyclodextrin, and bounds with 2-Hydroxy-Benzoic Acid. Substantial differences were also observed in the relative abundance of the top ten bacterial and fungal orders, and biomarker genera representatives. In cultivated orchards, there was a significant decrease in the relative abundance of many bacterial taxa such as e.g. Rhizobiales, Burkholderiales, Vivinamibacterales, and fungal taxa including Eurotiales, and Saccharomycetales. Notably, no significant differences were noted for Bacillus abundance among tested management practices. Forests favored Trichoderma abundance the most among tested practices (relative abundance 0.05 %). In turn, Trichoderma representatives were revealed as biomarker genera in gardens with animals. Williamsia representatives, as found in uncultivated orchards were suggested to be a biomarker of less disturbance, resulting from area restoration. Overall, the study discussed how different land management practices influence soil microbial communities and their functional roles, emphasizing the potential impacts of use on soil health and biodiversity within its implications. The most important recommendation bullet points:•using Williamsia representatives as a soil biomarker microorganism to indicate successful area restoration processes in apple orchards,•using d-Glucosaminic Acid metabolic stress test to reveal early difficulties in controlling fungal pathogens in soil microbial communities,•promoting diverse plant covers and reasonably reduced agrochemical inputs in apple orchards for enhancing soil microbial resilience,•preserving habitats of wild apple trees in forests or green belts, as these environments exhibit reduced metabolic stress and support the occurrence of fungi like Trichoderma, in the context of locations from which such isolates should be sought for further biocontrol use.

The geographies of apple cultivation: tracing the origins and dispersal of the wild apple Malus sieversii via the Silk Road to the Kashmir Valley

ABSTRACT The production and distribution of food is an important aspect of agricultural geography. Tracing the origin of crops is vital to ensuring future food security, especially in the context of geopolitical and climate change. Drawing on a case study of Kashmir, this article demonstrates how evidence from archaeobotany and ancient writings can be used to trace the geographies of apple domestication, cultivation and distribution. In doing so, it draws attention to the significance of trade networks; in this case those historically supported by the Silk Road.

Malus sieversii: a historical, genetic, and conservational perspective of the primary progenitor species of domesticated apples

<i>Malus sieversii</i>: a historical, genetic, and conservational perspective of the primary progenitor species of domesticated apples

Balancing the nutrient needs: Optimising growth in Malus sieversii seedlings through tailored nitrogen and phosphorus effects.

The impact of nitrogen (N) and phosphorus (P) on the physiological and biochemical processes crucial for tree seedling growth is substantial. Although the study of plant hydraulic traits in response to N and P is growing, comprehensive research on their combined effects remains limited. Malus sieversii, a key ancestral species of modern apples and a dominant species in Xinjiang's Tianshan wild fruit forest, is witnessing a decline due to climate change, pests and diseases, compounded by challenges in seedling regeneration. Addressing this, a 4-year study was conducted to determine the optimal fertilisation method for it. The experiment explored varying levels of N (N10, N20 and N40) and P (P2, P4 and P8), and their combined effects (N20Px: N20P2, N20P4, N20P8; NxP4: N10P4, N20P4 and N40P4), assessing their impact on gas exchange, hydraulic traits, and the interplay among functional traits in Tianshan Mountains' M. sieversii seedlings. Our study revealed that All N-inclusive fertilisers slightly promoted the net photosynthetic rate. N10 significantly increasing leaf hydraulic conductivity. All P-inclusive fertilisers adversely affected hydraulic conductivity. P8, N20P4 and N20P8 notably increased seedlings' vulnerability to embolism. Seedlings can adaptively adjust multiple functional traits in response to nutrient changes. The research suggests N10 and N20 as the most effective fertilisation treatments for M. sieversii seedlings in this region, while fertilisation involving phosphorus is less suitable. This study contributes valuable insights into the specific nutrient needs of it, vital for conservation and cultivation efforts in the Tianshan region.

Climate Change Impact on Three Important Species of Wild Fruit Forest Ecosystems: Assessing Habitat Loss and Climatic Niche Shift

As global biodiversity hotspots, wild fruit forests play key ecological functions, providing essential ecosystem services such as carbon storage, soil retention, and water conservation, and support food chains and biodiversity conservation through key species. Climate change, with rising temperatures and altered precipitation patterns, threatens wild fruit forests by reducing the habitats and numbers of key species, potentially turning these ecosystems from carbon sinks to sources and diminishing overall biodiversity and ecosystem services. However, research on how these changes affect important species’ habitats and carbon dynamics remains insufficient. To address this, we analysed habitat suitability for three important species (Prunus armeniaca L., Malus sieversii, and Prunus ledebouriana (Schltdl.) Y.Y.Yao with the aim of informing conservation strategies. We used biomod2 to integrate environmental and species data using six methods, encompassing 12 models. We predicted overlapping geographical distributions of three species, analysing their ecological niches and environmental interactions using global datasets to understand their adaptations. This analysis revealed ecological niche shifts and reductions in resource utilisation in both current and future scenarios. Their distribution centres will move poleward under the influence of bioclimatic factors and human activities. In conclusion, protecting P. armeniaca, M. sieversii, and P. ledebouriana is essential for the conservation and overall health of wild fruit forest ecosystems. This study provides new insights into climate change, habitat loss, informing conservation and resilience strategies.

Spatiotemporal patterns of leaf nutrients of wild apples in a wild fruit forest plot in the Ili Valley, China

Malus sieversii, commonly known as wild apples, represents a Tertiary relict plant species and serves as the progenitor of globally cultivated apple varieties. Unfortunately, wild apple populations are facing significant degradation in localized areas due to a myriad of factors. To gain a comprehensive understanding of the nutrient status and spatiotemporal variations of M. sieversii, green leaves were collected in May and July, and the fallen leaves were collected in October. The concentrations of leaf nitrogen (N), phosphorus (P), and potassium (K) were measured, and the stoichiometric ratios as well as nutrient resorption efficiencies were calculated. The study also explored the relative contributions of soil, topographic, and biotic factors to the variation in nutrient traits. The results indicate that as the growing period progressed, the concentrations of N and P in the leaves significantly decreased (P < 0.05), and the concentration of K in October was significantly lower than in May and July. Throughout plant growth, leaf N–P and N–K exhibited hyperallometric relationships, while P–K showed an isometric relationship. Resorption efficiency followed the order of N < P < K (P < 0.05), with all three ratios being less than 1; this indicates that the order of nutrient limitation is K > P > N. The resorption efficiencies were mainly regulated by nutrient concentrations in fallen leaves. A robust spatial dependence was observed in leaf nutrient concentrations during all periods (70.1–97.9% for structural variation), highlighting that structural variation, rather than random factors, dominated the spatial variation. Nutrient resorption efficiencies (NRE, PRE, and KRE) displayed moderate structural variation (30.2–66.8%). The spatial patterns of nutrient traits varied across growth periods, indicating they are influenced by multifactorial elements (in which, soil property showed the highest influence). In conclusion, wild apples manifested differentiated spatiotemporal variability and influencing factors across various leaf nutrient traits. These results provide crucial insights into the spatiotemporal patterns and influencing factors of leaf nutrient traits of M. sieversii at the permanent plot scale for the first time. This work is of great significance for the ecosystem restoration and sustainable management of degrading wild fruit forests.

Comparative analysis on natural variants of fire blight resistance protein FB_MR5 indicates distinct effector recognition mechanisms

FB_MR5 is an NLR protein identified from wild apple species Malus × robusta 5 conferring disease resistance to bacterial fire blight. FB_MR5 (hereafter MrMR5) recognizes the cysteine protease effector EaAvrRpt2 secreted from the causal agent of bacterial fire blight, Erwinia amylovora. We previously reported that MrMR5 is activated by the C-terminal cleavage product (ACP3) of Malus domestica RIN4 (MdRIN4) produced by EaAvrRpt2-directed proteolysis. Here, we show that MbMR5 from a wild apple species Malus baccata share 99.4% amino acid sequence identity with MrMR5. Surprisingly, transient expression of MbMR5 in Nicotiana benthamiana showed auto-activity in contrast to MrMR5. Domain swap and mutational analyses revealed that one amino acid polymorphism in the MbMR5 CC domain is critical in enhancing auto-activity. We further demonstrated that MrMR5 carrying seven amino acid polymorphisms present in MbMR5 is not activated by MdRIN4 ACP3 but recognizes AvrRpt2 without MdRIN4 in Nicotiana benthamiana. Our findings indicate that naturally occurring polymorphisms of MR5 natural variants can confer its cell death-inducing activity and the effector recognition mechanism likely due to altered compatibility with RIN4.

Pruning can recover the health of wild apple forests attacked by the wood borer Agrilus mali in central Eurasia

Pruning can recover the health of wild apple forests attacked by the wood borer Agrilus mali in central Eurasia

Pathogenicity evaluation of Cytospora species in 13 apple (Malus domestica) varieties and wild apple (Malus sieversii) in Xinjiang, China

AbstractApples are a popular fruit worldwide and are cultivated in the Xinjiang Uygur Autonomous Region (XUAR) in China. Cytospora canker disease is a serious problem in the apple industry in China. Pathogenicity of all historically recorded Cytospora species from Malus hosts was tested on the leaves and breaches of 13 varieties of apple (Malus domestica) and wild apple (Malus sieversii). Cytospora leucosperma and Cytospora pruinopsis were the most aggressive pathogens on apple and wild apple trees in Xinjiang. Meanwhile, Cytospora leucostoma, Cytospora nivea and Cytospora parakantschavelii were the least aggressive. In addition, Fuji and Golden Delicious were the most Cytospora canker disease‐resistant apple varieties and are recommended for future promotion. However, Sitagan and Erzizi were readily infected by Cytospora. Our findings provide a theoretical basis for field disease epidemic monitoring and tailoring disease prevention and control strategies based on the disease resistance of the variety and the aggressiveness of the pathogen.

Construction of a Membrane Yeast Two-Hybrid Library and Screening of MsPYR1-Like Interacting Proteins in Malus sieversii.

To investigate the biological effects of the ABA receptor pyrabactin resistance 1-like (PYR1-like) in Malus sieversii seeds, the proteins interacting with MsPYR1-like were screened by the membrane yeast two-hybrid library based on the split-ubiquitin system, and to construct the bait vector pBT3-SUC-PYR1 for Malus sieversii cDNA library, which had no self-activating effect on the yeast cells of the pPR3-N membrane yeast two-hybrid library. The library titer assay showed that it could meet the requirements for membrane yeast two-hybrid library screening. After sequencing, GenBank database blast, and yeast rotary validation, 28 candidate proteins interacting with MsPYR1-like were obtained, including ribosomal proteins, late embryogenesis abundant proteins, F-actin-capping proteins, phytochrome-interacting proteins, low-temperature-inducible 65kDa protein-like, senescence-associated, PP2C and SnRK2 family members, and unknown proteins. Gene ontology analysis of the interaction proteins was related to plant hormone response and negative regulation of seed germination, overexpression of MsPYR1-like in Arabidopsis negatively regulates seed germination, and the study of the biological roles of MsPYR1-like interacting proteins lays the foundation for revealing the lifting of seed dormancy in Malus sieversii.

In Vitro Approbation of Microbial Preparations to Shield Fruit Crops from Fire Blight: Physio-Biochemical Parameters.

The need for the increasing geographical spread of fire blight (FB) affecting fruit crops to be addressed led to large-scale chemicalization of the environmental matrices and reduction of plant productivity. The current study aimed to assess the effects of novel biopreparations at different exposure durations on photosynthetic pigment content and antioxidant enzyme activity in leaves of apple and pear varieties with varying levels of resistance to FB. Biopreparations were formulated from a cultural broth containing Lacticaseibacillus paracasei M12 or Bacillus amyloliquefaciens MB40 isolated from apple trees' phyllosphere. Aseptic leaves from blight-resistant (endemic Malus sieversii cv. KG10), moderately resistant (Pyrus pyraster cv. Wild), and susceptible (endangered Malus domestica cv. Aport and Pyrus communis cv. Shygys) varieties were employed. The impact of biopreparations on fruit crop antioxidant systems and photosynthetic apparatuses was investigated in vitro. Study results indicated that FB-resistant varieties exhibit enhanced adaptability and oxidative stress resistance compared to susceptible ones. Plant response to biopreparations varied based on the plant's initial FB sensitivity and exposure duration. Indeed, biopreparations improved the adaptive response of the assimilation apparatus, protein synthesis, and catalase and superoxide dismutase activity in susceptible varieties, suggesting that biopreparations have the potential for future commercialization to manage FB in fruit crops.

Physiological Studies and Transcriptomic Analysis Reveal the Mechanism of Saline-Alkali Stress Resistance of Malus sieversii f. niedzwetzkyan

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.

Evaluation of antioxidant potential of crude extract of metabolites from endophytic fungi isolated from &lt;i&gt;Annona senegalensis&lt;/i&gt; Pers

Degenerative diseases resulting from free radicals can be prevented through the use of natural antioxidants. This study aimed to assess the free radical scavenging potential of the underexplored endophytic fungal metabolites from Annona senegalensis Pers (wild custard apple). Freshly collected root, midrib, and leaf tissues were planted on malt extract agar following surface sterilization with 70% ethanol for 3 minutes, washed twice with distilled water, immersed in a sodium hypochlorite solution (4%) for 5 minutes, and subsequently rinsed with sterile water. Seven endophytic fungi were isolated, and the antioxidant activity of their metabolites was evaluated using the 2,2- diphenyl-1-picrylhydrazyl free radical scavenging method, with ascorbic acid serving as a reference antioxidant. The crude extract from the seven endophytic fungal isolates demonstrated concentration- dependent antioxidant activities. LB2 exhibited a strong antioxidant activity with an IC50 value of 38.15 μg/ml. Extracts from RT1 and LB1 showed antioxidant activities, with the highest inhibition of 60% and 65%, respectively, observed at a concentration of 1000 μg/ml. This was compared favorably to ascorbic acid, which exhibited 94% inhibition. This indicates that the crude extract from endophytic fungi isolated from A. senegalensis possesses free radical scavenging properties. Further purification and elucidation of the crude extract will reveal the bioactive compounds responsible for the antioxidant activities.

Safety assessment of crude aqueous methanol extract of Annona senegalensis stem bark: acute and sub-chronic toxicity studies.

Annona senegalensis Pers., (wild custard apple), is a shrub used traditionally to treat respiratory and skin diseases. Previous studies have demonstrated its anti-malaria, anti-snake envenomation and anti-cancer activities. However, its toxicological profile remains limited, particularly in male and female animals. This study aims to evaluate the safety of crude aqueous methanol extract of Annona senegalensis stem bark (AMEAS) through acute and sub-chronic toxicity studies. The stem bark of A. senegalensis was collected, air-dried, pulverized, and extracted using 70% methanol. Phytochemical screening, elemental analysis, and acute toxicity evaluation were carried out on AMEAS. Sub-chronic toxicity study was conducted on Wistar rats of both sexes at different doses administered orally for 28 days. Elemental analysis revealed the presence of heavy metals and essential mineral elements with the highest contents being calcium (59.88%) and potassium (25.39%). Acute toxicity testing showed no mortality up to 5000 mg/kg, suggesting an LD50 greater than 5000 mg/kg. In the sub-chronic toxicity study, no mortality or significant harmful effects were observed. The blood glucose decreased from 13.68 mMol/L at 250 mg/kg to 10.71 mMol/L at 1000 mg/kg, much lower than the distilled water group (17.06 mMol/L). In conclusion, the extract appeared to be well-tolerated, with no obvious adverse effects. AMEAS is rich in Calcium (Ca) and potassium (K). It has been shown to have LD50 greater than 5000 mg/kg and is assumed to be safe. On repeated use, AMEAS may cause hypoglycemia and weight loss which may be useful in managing diabetes and obesity respectively.