Search Results (20)

The WRKY transcription factor family is a significant family of plant transcription factors (TFs). Plant growth and development are often influenced by abiotic factors, such as salinity and low temperature. Numerous studies have demonstrated that WRKY TFs primarily influence plant responses to adversity. However, there are few studies on the role of WRKY genes in the stress responses of Malus baccata (L.) Borkh. We cloned the MbWRKY33 gene from Malus baccata for this research, and its roles in salt stress tolerance were analyzed. Phylogenetic tree analysis revealed that MbWRKY33 and PbWRKY33 have the highest homology. Subcellular localization revealed that MbWRKY33 was located within the nucleus. An analysis of tissue-specific expression showed that MbWRKY33 had relatively high expression levels in young leaves and roots. Moreover, Arabidopsis thaliana plants overexpressing MbWRKY33 exhibited stronger resistance to salt stress compared with the wild type (WT) and the unloaded line empty vector (UL). Under the treatment of 200 mM NaCl, transgenic Arabidopsis thaliana plants exhibited significantly higher activities of antioxidant enzymes like superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) than the control. In contrast, the WT and the UL lines had elevated levels of malondialdehyde (MDA) and reactive oxygen species (ROS). In addition, MbWRKY33 elevates transgenic plant resistance to salt stress by regulating the expression levels of AtNHX1, AtSOS1, AtSOS3, AtNCED3, AtSnRK2, and AtRD29a. Results indicated that MbWRKY33 in Malus might be linked to high-salinity stress responses, laying a foundation for understanding WRKY TFs’ reaction to such stress. Full article

Apple is an important horticultural crop, but various adverse environmental factors can threaten the quality and yield of its fruits. The ability of apples to resist stress mainly depends on the rootstock. Malus baccata (L.) Borkh. is a commonly used rootstock in Northeast China. In this study, it was used as the experimental material, and the target gene MbWRKY53 was screened through transcriptome analysis and Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR) after cold and drought treatment. Bioinformatics analysis revealed that this transcription factor (TF) belonged to the WRKY TF family, and its encoded protein was localized in the nucleus. RT-qPCR showed that the gene was more easily expressed in roots and young leaves and is more responsive to cold and drought stimuli. Functional validation in Arabidopsis thaliana confirmed that MbWRKY53 can enhance plant tolerance to cold and drought stress. Furthermore, by analyzing the expression levels of genes related to cold and drought stress in transgenic Arabidopsis lines, it was inferred that this gene can regulate the expression of stress-related genes through multiple pathways such as the CBF pathway, SOS pathway, Pro synthesis pathway, and ABA-dependent pathways, enhancing the adaptability of transgenic Arabidopsis to cold and drought environments. Full article

Wood can store carbon and help mitigate global climate change. Carbon density (CD), the basis for measuring and analyzing C storage, is the product of wood density (WD) and C concentration, which are dependent on wood structure, cellulose concentration (CC), hemicellulose concentration (HC), and lignin concentration (LC). However, little attention has been paid to the C concentration of cellulose, hemicellulose, and lignin, which are fundamental factors in C storage and affect the credibility of accurate CD estimates. In order to disentangle the CD drives, WD, C concentration, CC, HC, and LC of the branch, stem, and root were quantified for five Rosaceae species from temperate forests in Northeastern China. The species were Sorbus alnifolia (Sieb.et Zucc.) K. Koch, Pyrus ussuriensis Maxim., Malus baccata (L.) Borkh., Crataegus pinnatifida var. major N. E. Brown, and Padus racemosa (Linn.) Gilib. The WD, CC, HC, and LC differed among species and tree organs, with the highest variability for the HC. The structural carbon concentration (SCC) was lower than the organic carbon concentration (OCC) and even the Intergovernmental Panel on Climate Change (IPCC) default value of 45%, with a maximum deviation of 2.6%. CD differed dramatically among species and tree organs. Based on SCC calculations, the highest CD was found in Sorbus alnifolia root (0.27 × 10⁶ g/m³), while the lowest was found in Padus racemosa branch (0.22 × 10⁶ g/m³). The results suggest that when estimating CD accurately at species level, it is important to consider not only WD but also structural carbohydrates and lignin concentration, providing important information on C fluxes and long-term C sequestration for forests. The study findings provide valuable insights into CD variations among tree species and organs and are valuable for forest management and policy development to improve carbon sequestration. Full article

The living environment of plants is not static; as such, they will inevitably be threatened by various external factors for their growth and development. In order to ensure the healthy growth of plants, in addition to artificial interference, the most important and effective method is to rely on the role of transcription factors in the regulatory network of plant responses to abiotic stress. This study conducted bioinformatics analysis on the MbWRKY46 gene, which was obtained through gene cloning technology from Malus baccata (L.) Borkh, and found that the MbWRKY46 gene had a total length of 1068 bp and encodes 355 amino acids. The theoretical molecular weight (MW) of the MbWRKY46 protein was 39.76 kDa, the theoretical isoelectric point (pI) was 5.55, and the average hydrophilicity coefficient was −0.824. The subcellular localization results showed that it was located in the nucleus. After conducting stress resistance studies on it, it was found that the expression of MbWRKY46 was tissue specific, with the highest expression level in roots and old leaves. Low temperature and drought had a stronger induction effect on the expression of this gene. Under low temperature and drought treatment, the expression levels of several downstream genes related to low temperature and drought stress (AtKIN1, AtRD29A, AtCOR47A, AtDREB2A, AtERD10, AtRD29B) increased more significantly in transgenic Arabidopsis. This indicated that MbWRKY46 gene can be induced to upregulate expression in Arabidopsis under cold and water deficient environments. The results of this study have a certain reference value for the application of M. baccata MbWRKY46 in low-temperature and drought response, and provide a theoretical basis for further research on its function in the future. Full article

GABA (γ-aminobutyric acid) is found in plants and accumulates rapidly under stresses. However, the contributions of glutamic acid and a (Glu)-derived pathway and polyamines (PAs) catabolism pathway on GABA accumulation and the regulatory effects of exogenous putrescine (Put) on a GABA shunt under suboptimal low root-zone temperatures remain unknown. Our results showed that suboptimal low root-zone temperatures (treatment L) significantly increased GABA contents and GABA transaminase (GABA-T) activities. The contribution rate of the PAs catabolism pathway increased from 20.60% to 43.31%. Treatment L induced oxidative stress in Malus baccata Borkh. roots. Exogenous Put increased the contents of endogenous Put, spermine (Spm), and spermidine (Spd), promoted the transformation of PAs, increased the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), and decreased the contents of hydrogen peroxide (H₂O₂), superoxide anion (O₂∙⁻), and malondialdehyde (MDA). Meanwhile, contrasting results were observed after aminoguanidine (AG, an inhibitor of diamine oxidase) application. These findings revealed that the Glu-derived pathway is the main route of GABA synthesis. The contribution rate of the Pas catabolism pathway increased gradually with the extension of treatment time, and the treatment of exogenous Put significantly improved the tolerance of Malus baccata Borkh. Roots to suboptimal low temperature by regulating the transformation of Pas, GABA shunt, and the antioxidant system. Full article

Malus baccata (L.) Borkh. is an important wild species of Malus. Its rich variation types and population history are not well understood. Chloroplast genome mining plays an active role in germplasm identification and genetic evolution. In this study, by assembly and annotation, six complete cp genome sequences, ranging in size from 160,083 to 160,295 bp, were obtained. The GC content of stable IR regions (42.7%) was significantly higher than that of full length (36.5%) and SC regions (LSC-34.2%, SSC-30.4%). Compared with other Malus species, it was found that there were more sites of polymorphisms and hotspots of variation in LSC and SSC regions, with high variation sites including trnR/UCU-atpA, trnT/UGU-trnL/UAA, ndhF-rpl32 and ccsA-ndhD. The intraspecific and interspecific collinearity was good, and no structural rearrangement was observed. A large number of repeating elements and different boundary expansions may be involved in shaping the cp genome size. Up to 77 or 78 coding genes were annotated in the cp genomes of M. baccata, and high frequency codons such as UUA (Leu), GCU (Ala) and AGA (Arg) were identified by relative synonymous codon usage analysis. Phylogeographic analysis showed that 12 individuals of M. baccata clustered into three different groups with complex structure, whereas variant xiaojinensis (M.H. Cheng & N.G. Jiang) was not closely related to M. baccata evolutionarily. The phylogenetic analysis suggested that two main clades of different M. baccata in the genus Malus were formed and that I and II diverged about 9.7 MYA. In conclusion, through cp genome assembly and comparison, the interspecific relationships and molecular variations of M. baccata were further elucidated, and the results of this study provide valuable information for the phylogenetic evolution and germplasm conservation of M. baccata and Malus. Full article

As an excellent grafting material, Malus baccata (L.) Borkh is native to Liaoning, Jilin, Heilongjiang and other regions in China, with a strong adverse environmental adaptability. As a typical transcription factor, the NAC gene acts as a regulator in many molecular pathways responding to abiotic stress. However, research of NAC in the Malus baccata has just begun. In the present research, a new NAC transcription factor, MbNAC22, was obtained from the seedlings of Malus baccata, and its function in drought and salt treatments was studied by heterologous expression. The open reading frame of the MbNAC22 gene is 768, encoding 255 amino acids (aa). Through confocal microscopy, MbNAC22 was found to be located in the nucleus. The heterologous expression of MbNAC22 in Arabidopsis showed that it enhanced the viability of Arabidopsis under drought and salt treatments. Under stresses, the chlorophyll content of the plants decreased, but the decline of the overexpressed-MbNAC22 Arabidopsis was relatively low. Through phenotypic observation and determination of stress-related physiological indicators, it was found that compared with WT Arabidopsis, overexpressed-MbNAC22 Arabidopsis had a higher tolerance to stresses. Under stresses, the overexpression of MbNAC22 positively regulated ion-transport-related genes (AtNHX1 and AtSOS1), the key genes of the ABA pathway (AtNCED3 and AtDREB2A), the proline synthesis gene (AtP5CS2) and the drought-induced gene (AtERD11), while the expression of the leaf senescence-associated gene (AtSAG21) and programmed cell death related gene (AtAEP1) was inhibited. Therefore, we speculate that MbNAC22 responds positively to drought and salt stresses by regulating the expression of stress-related genes. Full article

Malus baccata Borkh., an apple rootstock, is found to be damaged by oxidation at sub-low root-zone temperature. In previous studies, we have found that exogenous sucrose could alleviate oxidative damage and increase the indole acetic acid (IAA) in roots under sub-low temperature (L). However, the role of IAA in sucrose-induced tolerance to L remains unclear. A pot experiment was conducted to evaluate the effects of exogenous sucrose and IAA synthesis/transport inhibitors (2,3,5-triiodobenzoic acid, TIBA; 4-biphenylboronic acid, BBo) on growth, IAA levels, sugars, and the antioxidant system of M. baccata under L. The results showed that the L treatment decreased IAA contents by 23.69% (48 h) and induced significant increases in root contents of malondialdehyde (MDA) and reactive oxygen (ROS), along with increasing catalase (CAT), ascorbate peroxidase (APX), and glucose-6-phosphate dehydrogenase (G6PDH) activities, while superoxide dismutase (SOD) and monodehydroascorbate reductase (MDHAR) activities first increased (24 h) and then decreased (48 h), and glutathione reductase (GR) and peroxidase (POD) activities significantly decreased. The L treatment also decreased ascorbate/oxidized ascorbate (AsA/DHA), glutathione/oxidized glutathione (GSH/GSSG), and coenzyme II/oxidized coenzyme II (NADPH/NADP⁺) ratios. Furthermore, the L treatment increased the contents of sucrose, fructose, glucose and sorbitol in the roots and suppressed plant growth. Sucrose pretreatment significantly increased IAA contents (12.42%, 24 h and 14.44%, 48 h) and decreased MDA and ROS contents, which improved the activities of antioxidant enzymes other than APX and increased the contents of AsA, GSH, and NADPH, and increased sucrose, fructose, and sorbitol contents and promoted plant growth. However, the sucrose + TIBA or BBo treatments decreased IAA contents and attenuated or almost abolished the positive effects of exogenous sucrose under sub-low temperature. Our findings indicate that IAA is involved in the sucrose-induced regulation of the antioxidant system in M. baccata roots under sub-low temperature and we provided theoretical references for further study on the adaptability of apple roots to low temperature. Full article

Malus baccata (L.) Borkh. is one of the most widely used rootstocks in the apple-producing region of Northern China. However, in the early growing season, apple roots are often subjected to suboptimal low root-zone temperatures. The regulatory effects of exogenous γ-aminobutyric acid (GABA) on both the γ-aminobutyric acid shunt (GABA shunt) and the respiratory activity of roots under suboptimal low root-zone temperatures remain unknown. To explore the physiological basis for GABA alleviation of low-temperature stress in M. baccata Borkh. roots, the following treatments were examined: suboptimal low root-zone temperature (potted parts of the seedlings were maintained at 5 ± 0.5 °C; L); suboptimal low root-zone temperature + GABA (LG); and suboptimal low root-zone temperature + vigabatrin (VGB; LV), which is a specific active inhibitor of γ-aminobutyric acid transaminase (GABA-T). Each treatment was matched with a control (18 °C/8 °C day/night; CK) for comparison. Our results showed that the L treatment reduced the root vitality, increased malondialdehyde (MDA) content, promoted the accumulation of GABA, activated the GABA shunt, and inhibited the total root respiration rate (V_Total) by decreasing the respiratory rates of Embden–Meyerhof pathway (V_EMP) and tricarboxylic acid cycle (V_TCAC). The LG treatment significantly increased the content of endogenous GABA, accelerated the metabolism of the GABA shunt, enhanced root respiratory activity by increasing V_Total, V_EMP, V_TCAC, and increased the cytochrome pathway respiratory rate (V_CP), thus alleviating the damage of low root-zone temperature stress. Meanwhile, contrasting results were observed in the LV treatment. These findings revealed that exogenous GABA improved the tolerance of apple rootstocks to suboptimal low temperatures in early spring by regulating the GABA shunt and root respiratory activity. Full article

Apples are a major horticultural crop worldwide. Grafting techniques are widely utilized in apple production to keep the varieties pure. Interstocks are frequently used in Northern China to achieve intensive apple dwarfing cultivation. High-throughput sequencing was used to investigate differentially expressed genes in the phloem tissues of two different xenograft systems, M (‘Gala’/‘Mac 9’/Malus baccata (L.) Borkh.) and B (‘Gala’/Malus baccata (L.) Borkh.). The results showed that dwarfing interstocks could significantly reduce the height and diameters of apple trees while have few effects on the growth of annual branches. The interstocks were found to regulate the expression of genes related to hormone metabolism and tree body control (GH3.9, PIN1, CKI1, ARP1, GA2ox1 and GA20ox1), these effects may attribute the dwarf characters for apple trees with interstocks. Besides, the interstocks reduce photosynthesis-related genes (MADH-ME4 and GAPC), promote carbon (C) metabolism gene expression (AATP1, GDH and PFK3), promote the expression of nitrogen (N)-metabolism-related genes (NRT2.7, NADH and GDH) in rootstocks, and improve the expression of genes related to secondary metabolism in scions (DX5, FPS1, TPS21 and SRG1). We also concluded that the interstocks acquired early blooming traits due to promotion of the expression of flowering genes in the scion (MOF1, FTIP7, AGL12 and AGL24). This study is a valuable resource regarding the molecular mechanisms of dwarf interstocks’ influence on various biological processes and transplantation systems in both scions and rootstocks. Full article

Malus baccata (L.) Borkh is an apple rootstock with good drought and cold resistance. The ICE gene is a key factor in the molecular mechanisms of plant drought and cold resistance. In the present research, the function of drought- and cold-induced MbICE1 of Malus baccata was investigated in Arabidopsis. According to GFP fluorescence images, MbICE1 was determined to be a nuclear protein. The MbICE1 was transferred to Arabidopsis, showing enhanced tolerance to drought and cold stresses. Under drought and cold treatments, the transgenic Arabidopsis had higher chlorophyll content and free proline content than WT plants, but the Malondialdehyde (MDA) content and electrolyte leakage (EL) were lower than those of WT plants. In addition, drought and cold led to a large accumulation of ROS (H₂O₂ and O²⁻) content in Arabidopsis, while overexpression of MbICE1 enhanced the antioxidant enzyme activity in Arabidopsis and improved the plant’s resistance to stresses. Moreover, the accumulation of MbICE1 promoted the expression of AtCBF1, AtCBF2, AtCBF3, AtCOR15a, AtCOR47 and AtKIN1 genes in Arabidopsis. These data indicate that MbICE1 is a key regulator of drought and cold and can be used as a backup gene for breeding Malus rootstocks. Full article

In the early growing season in northern China, suboptimal low root-zone temperatures is a common abiotic stress that impairs root function and leaf development in fruit trees. In this study, we investigate the physiological role of leaves in jasmonate metabolism and the capacity of scavenging reactive oxygen species in Malus baccata (L.) Borkh. roots under suboptimal low root-zone temperatures. In the presence of intact leaves, suboptimal low root-zone temperatures significantly increased allene oxide synthase (AOS), jasmonate-resistant 1 (JAR), and jasmonic acid carboxyl methyltransferase (JMT) activities and transcription in jasmonate biosynthesis. Meanwhile, elevated endogenous jasmonic acid (JA), methyl jasmonate (MeJA), and jasmonate-isoleucine (JA-Ile) contents were also observed, as were significantly decreased glutathione reductase and dehydroascorbate reductase activities and AsA/DHA and GSH/GSSG ratios. Conversely, leaf removal substantially reduced AOS, JMT, and JAR activities and transcription at most time points and JA (6–24 h), MeJA (1–24 h), and JA-Ile (1–24 h) levels in roots, affecting key enzymes in the AsA–GSH cycle and the AsA/DHA and GSH/GSSG ratios in response to low-temperature treatment, as a result of a significant increase in malondialdehyde content. Thus, leaves are crucial for jasmonate metabolism in roots under suboptimal low root-zone temperatures, with leaf removal exacerbating root oxidative stress by altering JA signaling and AsA–GSH cycle activity. Full article

Understanding the different physiological responses of Malus species under salt stress in the seedling stages will be useful in breeding salt-tolerant dwarfing apple rootstocks. Seedlings of Malus Zumi (Mats.) Rehd. (M. zumi), Malus sieversii (Led.) Roem. (M. sieversii), and Malus baccata (L.) Borkh. (M. baccata) were treated with solution of 0, 0.20%, 0.40%, and 0.60% salinity. Physiological parameters of their leaves and roots were measured at 0 d, 4 d, 8 d and 12 d after salinity treatments. Superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), malondialdehyde (MDA), solution protein (SP), and proline (PRO) initially increased and then decreased. The activities and contents of these parameters were higher in the 0.40% and 0.60% NaCl treatments than in the 0.20% treatment and in the 0% control. M. zumi was the most resistant to salt stress, showing the lowest content of MDA in the leaves and roots, which increased slightly under salt stress. M. baccata had the highest increase in both the content and proportion of MDA. High enzyme activity was shown to play an important role in the salt resistance of M. zumi. Moreover, it can be speculated that there are other substances that also play a major role. We found that osmotic regulation played a key role in response to salt stress for M. baccata even though it was sensitive to salt stress. For M. sieversii, both the osmotic regulation and enzymatic antioxidants were observed to play a major role in mitigating salt stress. Full article

Ultraviolet (UV) irradiation induces ROS production, which activates activator protein (AP)-1 and nuclear factor (NF)-κB signaling and downstream molecules, ultimately triggering the generation of matrix metalloproteinases (MMPs) and degradation of collagen. The aim of this study was to investigate the protective effect of methanol extract from Malus baccata (L.) Borkh (Mb-ME) against aging. DPPH and ABTS assays showed that Mb-ME had a significant antioxidant capacity. Flow cytometry results indicated that Mb-ME attenuated UVB and H₂O₂-stimulated apoptosis and reactive oxygen species (ROS) generation. RT-PCR analysis in HaCaT and HDF cells suggested that Mb-ME treatment blocked the expression of MMPs, COX-2, IL-1β, IL-6, HYALs, and p53 while promoting the levels of TGM1, FLG, HASs, Sirt1, and Col1A1. Mechanically, Mb-ME inhibited the phosphorylation of MAP kinases and NF-κB signaling. Overall, these results strongly suggest that Mb-ME can be developed as an antiaging therapy. Full article

CBF transcription factors (TFs) are key regulators of plant stress tolerance and play an integral role in plant tolerance to adverse growth environments. However, in the current research situation, there are few reports on the response of the CBF gene to Begonia stress. Therefore, this experiment investigated a novel CBF TF gene, named MbCBF2, which was isolated from M. baccata seedlings. According to the subcellular localization results, the MbCBF2 protein was located in the nucleus. In addition, the expression level of MbCBF2 was higher in new leaves and roots under low-temperature and high-salt induction. After the introduction of MbCBF2 into Arabidopsis thaliana, the adaptability of transgenic A. thaliana to cold and high-salt environments was significantly enhanced. In addition, the high expression of MbCBF2 can also change many physiological indicators in transgenic A. thaliana, such as increased chlorophyll and proline content, superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activity, and reduced malondialdehyde (MDA) content. Therefore, it can be seen from the above results that MbCBF2 can positively regulate the response of A. thaliana to low-temperature and osmotic stress. In addition, MbCBF2 can also regulate the expression of its downstream genes in transgenic lines. It can not only positively regulate the expression of the downstream key genes AtCOR15a, AtERD10, AtRD29a/b and AtCOR6.6/47, related to cold stress at low temperatures, but can also positively regulate the expression of the downstream key genes AtNCED3, AtCAT1, AtP5CS, AtPIF1/4 and AtSnRK2.4, related to salt stress. That is, the overexpression of the MbCBF2 gene further improved the adaptability and tolerance of transgenic plants to low-temperature and high-salt environments. Full article