In order to explore the application of coffee protein in food processing, non-fat green coffee powder was used as the raw material. Different ultrasonic powers (0 W, 200 W, 400 W, 600 W and 800 W) effect of synergistic interaction with deep eutectic solvent method and alkali method on composition (SDS-PAGE), structural properties (particle size distribution, Fourier transform infrared spectroscopy, circular dichroism spectrum, fluorescence spectroscopy and scanning electron microscopy) and functional properties (solubility, water absorption, oil absorption and emulsification) of coffee protein were studied. The average particle size of coffee protein (DES) extracted by deep eutectic solvent (12.30 μm) was significantly smaller than that of coffee protein (AEP) extracted by alkali method (113.67 μm). The solubility of DES was higher under acidic conditions, and the solubility of AEP was higher under alkaline conditions. DES had better water and oil absorption than AEP. After ultrasonic pretreatment, the results of coffee protein electrophoresis, Fourier infrared spectrum, circular dichroism spectrum and fluorescence spectroscopy showed that different ultrasonic power pretreatment did not cause significant changes in protein molecular weight distribution, but it affected the secondary and tertiary structure of proteins. The surface microstructure of DES showed that the surface structure was loose and porous, and the surface of AEP was tight and flat. After ultrasonic pretreatment, the microstructure of AEP became loose and porous, the texture became dispersed, and more irregular fragments were presented. The surface of DES changed slightly, and showed more small-particle porous disordered structure when treated at 400 W and 600 W for 15 min. The microstructure changes could affect the chemical and physical properties and functional properties of proteins. When the ultrasonic power was 400 W, the particle size of DES decreased significantly, and the particle size parameters D_[3,2] and D_[4,3] decreased from 10.50 μm, 30.43 μm to 7.89 μm and 11.27 μm, respectively. The solubility increased significantly, and the solubility increased from 47% and 71% to 75% and 81% respectively under acidic and alkaline conditions. In addition, ultrasonic pretreatment could improve the water absorption and emulsification activity of coffee protein, which are important functional properties of proteins in food applications. The results show that the modification of coffee protein by ultrasonic technology is effective, and this study could provide a theoretical basis for the application of coffee protein in food processing.

The purpose of this study was to provide a scientific basis for the classification, breed selection and targeted development and utilization of different germplasm of Aquilaria sinensis. The samples of agarwood from six different germplasms were studied: Aquilaria sinensis ‘Haixiang 1’, A. sinensis ‘Haixiang 2’, A. sinensis ‘Haixiang 3’, Piyou type, Big leaf type and Nangpao type. Gas chromatography-mass spectrometry (GC-MS) was used to study the composition of the agarwood samples extracted by ultrasonic ether method. The volatile components of agarwood produced by the above six germplasms were higher in sesquiterpenoids and lower in chromones, The volatile components of six germplasms were quite different, except two components in common, namely butylated hydroxytoluene and 2,2'-methylenebis[6-(1,1-dimethylethyl)-4-methyl-phenol]. The number of sesquiterpenoids (10-26 kinds) and the total relative percentage content (25.39%-87.50%) were higher, and the main component was baimuxinal, among which A. sinensis ‘Haixiang 3’ had the highest amount and content. The quantity (1-3 species) and total relative percentage content (4.66%-37.32%) of chromones were low, mainly 2-(2-phenylethyl) chromones, among which A. sinensis ‘Haixiang 1’had the highest quantity and content, while A. sinensis ‘Haixiang 3’, Big leaf type and Nangpao type were not detected. The agarwood from A. sinensis ‘Haixiang 1’, A. sinensis ‘Haixiang 2’, A. sinensis ‘Haixiang 3’, Piyou type, had relatively rich aromatic components (baimuxinal,7α-H-9(10)-ene-11,12-epoxy-8-oxoeremophilane, karanone and so on). The agarwood produced by A. sinensis ‘Haixiang 2’, A. sinensis ‘Haixiang 3’, Piyou type and Nangpao type contained many sesquiterpenoids, such as agarospirol and (-) jinkoh-eremol. The content of benzylacetone in A. sinensis ‘Haixiang 1’, A. sinensis ‘Haixiang 2’ agarwood was high. 7α-H-9(10)-ene-11,12-epoxy-8-oxoeremophilane and 6,7-dimethoxy-2-(2-phenylethyl) chromone in the agarwood produced by A. sinensis ‘Haixiang 1’, A. sinensis ‘Haixiang 3’ and Big leaf type were high. The types and contents of volatile components of agarwood derived from the six different germplasms showed great difference, which could be developed according to the difference of active components.

Natural rubber, a crucial industrial raw material worldwide, and a significant part of the income for Laotian farmers, is influenced by alternative planting policies and fluctuations in rubber prices. Aaccurate and rapid monitoring of local artificial rubber plantation is of crucial significance for promoting the sustainable development of the rubber industry. Most research has focused on the extraction of rubber plantation information in small areas, typically utilizing single or dual-temporal image classification, which fails to capture the long-term expansion characteristics of rubber forests. To investigate the long-term, large-scale expansion features of rubber plantation, this study selected a continuous thirty-year Landsat time series imagery from 1990 to 2020. The LandTrendr algorithm was employed to extract changes in rubber plantation across Laos over the three decades. International rubber futures prices and terrain data were also utilized for spatiotemporal change analysis. The selected feature combination, after optimization, can better extract rubber forest distribution. The overall accuracy of rubber forest distribution extraction in 2020 was 89.85%, with a Kappa coefficient of 0.82, revealing Laos’ rubber plantation area to be 321 000 hectares in 2020. The overall accuracy of rubber forest expansion change monitoring using the LandTrendr algorithm and secondary classification was 92.44%, with a Kappa coefficient of 0.82. The expansion of artificial rubber forests in Laos was closely related to the rubber futures market. Through terrain factor analysis, it is deduced that rubber forests are suitable for planting in low-altitude, gently sloping, and sunny areas. This study effectively monitors changes in rubber forests at the national level over an extended period, providing valuable data support for the development of the rubber industry and policy control.

The effect of bacterial wilt on the fertility of different textures of latosol in Hainan province (sand from Wencheng town, Wenchang city; loam from Dacheng town, Danzhou city; clay from Yongfa town, Chengmai county) was investigated. After the occurrence of bacterial wilt, the disease incidence, physicochemical indicators including pH, organic matter (OM), available nitrogen (AN), available phosphorus (AP), available potassium (AK), soil porosity (SP), bulk density (BD), mass water content (MWC) as well as enzyme activity indicators including catalase (CAT), peroxidase (POD), sucrase (SC), urease (UE) and acid phosphatase (ACP) of the soil inoculated with Ralstonia solanacearum and blank control were compared to analyze the effects of bacterial wilt on various fertility indicators. Spearman correlation analysis and redundancy analysis (RDA) were used to determine the significant indicators affected by bacterial wilt in different soil textures. The results showed that the disease incidence and severity disease index in clay were the highest among the three soil textures, while those of loam were the lowest. In the inoculation treatments, pH, AP, SP and POD of the three soil textures were lower than those of the control, while AN, AK, MWC and UE of clay inoculation treatment were higher than those of the clay control. CAT, ACP, MWC and BD of loam inoculation treatment were higher than those of the loam control. Spearman correlation analysis results showed that OM content, AP content, SP and POD of the three texture soils were negatively correlated with disease incidence; ACP, pH and disease incidence in WCS were significantly negatively correlated (P<0.05) with disease incidence; CAT and UE were significantly positively correlated (P<0.05) with disease incidence. In loam, pH, OM, POD and UE was significantly negatively correlated with disease incidence (P<0.05); while CAT and ACP were significantly positively correlated with disease incidence (P<0.05); SP and POD in clay were significantly negatively correlated with disease incidence (P<0.05); however, AK content in clay was significantly positively correlated with disease incidence (P<0.05). RDA results showed that the SP, OM content and AN content in the control and inoculation treatments were highly correlated with UE, while both SP and AK content in the control and inoculation treatments were highly correlated with ACP and CAT respectively. After the occurrence of bacterial wilt, most fertility indicators of control and inoculation treatment of sand and loam were lower than those in clay. However, soil fertility level was not a sufficient condition for bacterial wilt, but most soil fertility indicators were reduced due to the outbreak of bacterial wilt. The lower disease incidence and disease index of tomato bacterial wilt in sand than in clay may be due to the higher AP content. The reason why disease incidence and disease index of tomato bacterial wilt in loam is lower is perhaps that low pH conditions are unsuitable for the survival of bacterial wilt. Clay with the most severe bacterial wilt disease situation including pH, nitrogen level and mass water content was suitable for the growth and reproduction of bacterial wilt. The relative abundance of potential beneficial microbial communities in clay may also be lower than sand and loam. Therefore, the control measures of bacterial wilt in different soil textures should be adapted to local conditions. Meanwhile, the relevant conclusions of this study would provide a theoretical basis for establishing the control system of bacterial wilt in Hainan province in the future.

AT-hook nuclear localized proteins (AHLs) are small DNA-binding protein motifs that play an important role in plant growth and development, organ construction, stress and hormone signaling response. In this study, MeAHL17 was cloned from cassava cultivar SC8. Bioinformatics methods were used to analyze MeAHL17 promoter, protein physicochemical properties, prediction of conserved functional domain and sequence comparison. The results showed that the open reading frame length of MeAHL17 gene was 939 bp, encoding a protein with 312 amino acids, its theoretical isoelectric point was 6.89, its molecular formula was C₁₄₂₀H₂₂₁₅N₄₁₅O₄₅₁S₁₁, and its molecular weight was 32.669 38 kDa. The total number of positively charged amino acid residues (Lys+Arg) was 23, the total number of negatively charged amino acid residues (Asp+Glu) was 24, the fat coefficient was 57.47, the total average hydrophilic coefficient (GRAVY) was -0.491 and the instability coefficient was 58.06. The protein contains no signal peptide, is located in the cell membrane, has no transmembrane structure, contains 5 glycosylation sites and 45 phosphorylation sites, and is an unstable hydrophilic acidic protein MeAHL17 contained AT-hook conserved core sequences and PPC domain conserved core sequences, which was consistent with the typical structural characteristics of AT-hook family. Through subcellular localization and transcriptional activity analysis experiments, MeAHL17 was proved to be a nucleus localized transcription factor with transcriptional activity. Tissue expression pattern analysis showed that MeAHL17 was mainly expressed in somatic embryo, fibrous root and callus. Different hormone treatments showed that MeAHL17 was induced by ethylene (ACC), methyl jasmonate (JA) and growth hormone (IAA), which suggested that MeAHL17 might be involved in ethylene, methyl jasmonate and growth hormone signaling pathway. Abiotic stress treatment showed that MeAHL17 was responsive to drought and salt stress. This study preliminarily identified the important role of MeAHL17 in growth and development, hormone signaling and stress, and provided theoretical basis and reference for further study of its function.

Powdery mildew caused by Erysiphe quercicola is a significant disease in China’s natural rubber industry, posing a severe threat to latex production. With the continuous advancement of predictive and forecasting technologies, the combination of qPCR detection and spore capture techniques holds promise as an efficient method for predicting the prevalence of powdery mildew. In this study, we focused on the microscale extraction of DNA from rubber tree powdery mildew spores. We compared various pre-treatment methods, different types of grinding beads with varying materials, and different types of grinding machines to identify the most suitable spore sample grinding system. Furthermore, we compared the efficiency of the CTAB method with several commercial extraction kits. Ultimately, we established a feasible microscale DNA extraction method for spores: first, the powdery mildew spore water solution subjected to 4 ℃ultrasonication for 10 min, subsequently, 2 mm zirconia beads (0.2 g), glass beads (0.4-0.6 mm, 0.2 g), and 0.2 mm zirconia beads (0.2 g) added, tissue grinder 70 Hz at 4 ℃, grind 40 s, stop 10 s, 20 cycles, finally, DNA extracted using the CTAB method. This method effectively and stably extracts DNA from 10¹ spores/mL rubber tree powdery mildew spores, without being affected by complex field conditions, and demonstrates high sensitivity and specificity. It provides a reliable theoretical basis and an effective technical approach for the prediction and forecasting of rubber tree powdery mildew.

Cassava mosaic disease (CMD) poses a serious threat and danger to cassava production in China and the world, and there is no effective control method and lack of resistant materials, causing serious economic losses, and is considered one of the most threatening viral diseases. Sri Lankan cassava mosaic virus (SLCMV) is one of the major pathogens causing cassava mosaic disease. RNA silencing is an innate antiviral immune response mechanism in plants and animals, and post transcriptional gene silencing (PTGS) is a conserved gene expression regulatory mechanism in eukaryotes, and is one of the important antiviral immune mechanisms. Suppressor of gene silencing 3 (SGS3) is a key protein of the PTGS pathway and plays a critical role in host defense against viral invasion. To counteract this host immune response, viruses usually encode viral suppressors of RNA silencing (VSRs) to counteract the viral defense mechanism of host PTGS, and interaction with host proteins is one of the main mechanisms of action for the silencing suppressors to function. In order to investigate whether SLCMV AC2 inhibits the immune function of host PTGS by interacting with AtSGS3, we analyzed the immune function of host PTGS by using yeast two-hybrid (Y2H) system and bimolecular fluorescence complementation (BiFC) experiments, to analyze the interaction between AtSGS3 and SLCMV AC2. The results showed that SLCMV AC2 interacted with AtSGS3 protein in both yeast cells and plants. In subcellular co-localization experimental studies, we further demonstrated that SLCMV AC2 could co-localize with AtRDR6, a close partner of AtSGS3, after interacting with AtSGS3. The data suggest that SLCMV AC2 may affect the host autoimmune mechanism and enhance viral pathogenicity by interacting with siRNA-body, and that the interaction between SLCMV AC2 and AtSGS3 may be the molecular basis for the development of foliar symptoms in SLCMV-infected cassava, and the results of this study would provide a new theoretical basis for the subsequent in-depth analysis of the pathogenicity mechanism of SLCMV.

Banana Fusarium wilt is a fungal disease which is the most harmful and difficult to control in banana production in China. Due to the influence of banana Fusarium wilt, the banana planting area in China has been decreasing year by year in recent years, which has brought huge economic losses to the subtropical and tropical banana industries in China. The disease occurred because the vascular bundle of banana was invaded by Fusarium oxysporum f. sp. cubense (Foc), and its propagation speed was fast, which caused the yellowing of banana leaves and plant wilting. By comparing the genome and transcriptome sequences of Foc race 1 (Foc1, N2) and race 4 (Foc4, B2), a secreted protein FoSP1 located in the lineage-specific region with high expression was screened out. The results showed that the open reading frame of FoSP1 gene was 387 bp, encoding 129 amino acids, and its signal peptide cleavage site was located between the 20th and 21st amino acid residues, and the protein had no known domain and functional site, so it was inferred that FoSP1 was a new secreted protein. In order to study the biological function of the protein in Foc4 strain, the split-marker method was used to knock out the FoSP1 gene of B2 strain, and the phenotype analysis and pathogenicity determination of the correctly knocked-out mutant were carried out. The results showed that compared with wild-type B2 strain, there was no significant difference in the vegetative growth of FoSP1 gene knockout mutants, and they were insensitive to exogenous stresses such as NaCl, D-sorbitol and H₂O₂, but the conidial yield and germination rate of knockout mutants were significantly reduced, and their pathogenicity to Brazilian banana was significantly weakened. It is inferred that the secreted protein FoSP1 does not participate in the vegetative growth of B2 strain, but plays an important role in the process of sporulation and pathogenicity. This result lays a foundation for further study on the pathogenic mechanism of secreted proteins in the genome lineage-specific region of Foc4.

Tigernut (Cyperus esculentus L.) is an herbaceous oil crop uniquely accumulating a high level of oil in the underground tubers. Water, which accounts for approximately 85% of immature tubers, is essential for tuber development and metabolism. Plasma membrane intrinsic proteins (PIPs) represent the main channel that mediates the efficient water transport across the cell membrane. This study presents the characterization of CePIP2;1, a PIP gene highly abundant in tigernut tubers. This gene with three introns was shown to encode 288 amino acids with the theoretical molecular weight of 30.34 kDa, the isoelectric point of 8.60, the grand average of hydropathicity of 0.529, and the instability index of 29.60, implying it’s stable, basic and hydrophobic characteristics. The protein was shown to possess one conservative MIP (major intrinsic protein) domain, and presence of the F-H-T-R ar/R selectivity filter and Q-S-A-F-W Froger’s positions implies a putatively efficient water transport activity. Phylogenetic analysis revealed that CePIP2;1 clusters with OsPIP2;1, OsPIP2;2, and OsPIP2;3 with the similarities of 91.72%, 90.31% and 84.98%, respectively, supporting its assignment into the PIP2 group of the PIP subfamily. Subcellular localization analysis showed that CePIP2;1 was localized to the cell membrane of Nicotiana benthamiana leaves. Although CePIP2;1 was characterized as a dominant isoform in tubers, expression analysis indicated that its transcripts were relatively more in leaves, leaf sheaths, rhizomes, and roots with the most in leaves, in contrast to a comparable level to that in shoot apexes. Among four typical stages (S₁–S₄) of tuber development examined in this study, CePIP2;1 was shown to exhibit a bell-shaped expression pattern, peaking at S₂. These results laid a solid foundation for further uncovering the mechanism of water balance in tigernut.

This study was conducted to compare the difference of heat tolerance of different Oncidium cultivars, and to establish a system for evaluating identification indexes of Oncidium heat tolerance. Seven indexes, including the leaf thickness, leaf water content and high temperature semi-lethal temperature (HLT₅₀), of 38 common O. hybridum cultivars were measured, and the yellow leaf rate and heat injury grade (HIG) of different cultivars were observed under high temperature stress. The evaluation indexes of heat tolerance of Oncidium were screened by analyzing the correlation among different indexes and combining with principal component analysis, to establish the evaluation system of heat resistance of Oncidium cultivars. The results showed that the leaf thickness, leaf water content, HLT₅₀, yellow leaf rate and HIG could well reflect the response of Oncidium to high temperature stress, which could be taken as the index for evaluating Oncidium heat tolerance. The 38 cultivars could be classified into three categories: heat-tolerant cultivars (12 cultivars), moderate heat-tolerant cultivars (3 cultivars) and non-heat-tolerant cultivars (23 cultivars) by cluster analysis based on heat-tolerant index. The results of this study could provide a reference for material selection of Oncidium in hot area, and also provide parental resource for the breeding of Oncidium with high heat tolerance.