Alternative splicing of the PECTINESTERASE gene encoding a cell wall-degrading enzyme affects postharvest softening in grape

The firmness of table grape berries is a crucial quality parameter. Despite extensive research on postharvest fruit softening, its precise molecular mechanisms remain elusive. To enhance our comprehension of the underlying molecular factors, we initially identified differentially expressed genes(DEGs) by comparing the transcriptomes of folic acid(FA)-treated and water-treated(CK) berries at different time points. We then analyzed the sequences to detect alternatively spliced(AS) genes associated with postharvest softening. A total of 2,559 DEGs were identified and categorized into four subclusters based on their expression patterns, with subcluster-4 genes exhibiting higher expression in the CK group compared with the FA treatment group. There were 1,045 AS-associated genes specific to FA-treated berries and 1,042 in the CK-treated berries, respectively. Gene Ontology(GO) annotation indicated that the AS-associated genes in CK-treated berries were predominantly enriched in cell wall metabolic processes,particularly cell wall degradation processes. Through a comparison between treatment-associated AS genes and subcluster-4 DEGs, we identified eight genes, including Pectinesterase 2(VvPE2, Vitvi15g00704), which encodes a cell wall-degrading enzyme and was predicted to undergo an A3SS event. The reverse transcription polymerase chain reaction further confirmed the presence of a truncated transcript variant of VvPE2 in the FA-treated berries.Our study provides a comprehensive analysis of AS events in postharvest grape berries using transcriptome sequencing and underscores the pivotal role of VvPE2 during the postharvest storage of grape berries.

Identification,characterization,and verification of miR399 target gene in grape

The microRNA miR399 plays an important role in phosphorus signal transduction pathways in plants.Previously,miR399 was shown to be closely associated with berry ripening in grape(Vitis vinifera).The objective of the present study was to elucidate the evolutionary characteristics of the miR399 gene family in grape and to verify the cleavage effect on the target genes.Grape miR399s were identified by miRNA sequencing and retrieval from the miRBase database.The mature sequences and precursor sequences were subjected to phylogenetic analysis to reconstruct evolutionary trees,as well as secondary structure analysis of the precursor sequence,and prediction of target genes.The cisacting elements in the miR399 promoter were predicted and the cleavage effect of grape miR399b on its target genes was verified.The grape miR399 family comprised nine precursor sequences and nine mature sequences.The precursor sequences formed a typical and stable stem—loop structure.The minimum folding free energy ranged from-55.70 kcal·mol^(-1)to-37.40 kcal·mol^(-1).Multiple sequence alignment revealed that the miR399 family was highly conserved.The grape miR399 family was phylogenetically closely related to peach,apple,and citrus miR399s.Grape miR399s were predicted to target inorganic phosphate transporter 1—3,phospholipase D delta-like,and beta-glucuronosyltransferase.The cleavage effect of grape miR399b on the target genes was verified by means of a dual-luciferase assay and 5’RLM-RACE.Histochemical GUS staining showed that the promoter activity of miR399b was promoted by GA3treatment.

Experimental Study on the Attenuation Law of Vibration Wave Propagation in Natural Gas Wells in Coal-Gas Cross-Mining Area

Aiming at the safety distance between coal mining working face and natural gas wells in the cross-mining area of multiple mineral resources, the cross- mining area of gas and coal resources in the Ordos Basin is taken as the engineering background. An anti-collision early warning technology method based on vibration wave propagation attenuation monitoring is proposed to prevent collision accidents between road headers and natural gas wells. Through the steel pipe and steel pipe concrete knocking vibration test and underground digging vibration test, the research results show that: The exponential decay coefficients of the vibration wave in steel pipe, steel pipe concrete and coal rock respectively are 0.1, 0.1140 and 0.03, which are all in accordance with the exponential decay law, and the vibration wave firstly decays sharply and then decays slowly;the formula for the distance from the road header to the natural gas well was derived based on the vibration attenuation formula, to provide a new method for realizing the problem of precise and coordinated extraction by surface monitoring of the distance from down hole road headers to gas wells, collision prevention prediction and warning and prevention of collision of extraction equipment with gas wells.

Water-conserving mining influencing factors identification and weight determination in northwest China

Water-conserving mining is an effective way to alleviate the contradiction between fragile ecological environment and high-intensity coal mining in the arid and semi-arid region of northwest China. It needs to consider the engineering and geological conditions, hydrogeological conditions and mining methods of coal seams. From the three aspects, this paper systematically analyzes the influencing factors and establishes an identification model with multi-level structures. The model includes three primary factors (including the engineering and geological conditions, hydrogeological conditions and mining methods), nine secondary factors (including overlying strata thickness, aquiclude, mining parameters and etc.), sixteen third-tier factors (including the faults, aquiclude thickness and effective mining height and etc.) and twelve fourth-tier factors (including the fault throw exp on ent, aquiclude permeability and coal pillar sizes and etc.). On the basis, the analytic hierarchy process is used to build the judgment matrix and obtain the weight of each influencing factor. The results indicate that the overlying strata thickness, aquiclude and effective mining height are the most import a nt factors among the primary factors of engineering and geological conditions, hydrogeological conditions and mining methods, respectively. The research results could provide theoretical references for the water-conserving mining of coal resources in northwest China.