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Quantitative real-time PCR (qRT-PCR) has been emerged as an effective method to explore the gene function and regulatory mechanisms. However, selecting appropriate reference gene (s) is a prerequisite for obtaining accurate qRT-PCR results. Peach is one of important fruit in Rosaceae and is widely cultivated worldwide. In this study, to explore reliable reference gene (s) in peach with different types during fruit ripening and softening (S1–S4), nine candidate reference genes ( EF-1α , GAPDH , TBP , UBC , eIF-4α , TUB-A , TUB-B , ACTIN , and HIS ) were selected from the whole-genome data. Then, the expression levels of the nine selected genes were detected using qRT-PCR in three peach types, including ‘Hakuho’ (melting type), ‘Xiacui’ (stony hard type), ‘Fantasia’ and ‘NJC108’ (non-melting type) cultivars were detected using qRT-PCR. Four software (geNorm, NormFinder, BestKeeper and RefFinder) were applied to evaluate the expression stability of these candidate reference genes. Gene expression was characterized in different peach types during fruit ripening and softening stages. The overall performance of each candidate in all samples was evaluated. The Actin gene ( ACTIN ) was a suitable reference gene and displayed excellent stability in ‘Total’ set, ‘Hakuho’ samples, S3 and S4 fruit developmental stages. Ubiquitin C gene ( UBC ) showed the best stability in most independent samples, including ‘Fantasia’, ‘NJC108’, S2 sets. Elongation factor-1 α gene ( EF-1α ) was the most unstable gene across the set of all samples, ‘NJC108’ and S2 sets, while showed the highest stability in ‘Xiacui’ samples. The stability of candidate reference genes was further verified by analyzing the relative expression level of ethylene synthase gene of Prunus persica ( PpACS1 ) in fruit ripening and softening periods of ‘Hakuho’. Taken together, the results from this study provide a basis for future research on the mining of important functional genes, expression patterns and regulatory mechanisms in peach.

Automated peach picking in complex orchard environments faces challenges such as fruit overlap, occlusion by leaves and branches, and low efficiency in continuous localization, which require detection algorithms to achieve both high accuracy and real-time performance. To address these issues, this study proposes a lightweight and high-precision peach detection model named Peach-YOLO based on an improved YOLOv8n framework. First, a Receptive-Field Attention Convolution (RFAConv) module is introduced into the C2f structure of the backbone network to enhance feature representation in salient regions and suppress background interference. Second, a Convolution and Attention Fusion Module (CAFM) is integrated to further strengthen the extraction of key fruit features. In the neck network, a Coordinate Attention-guided high-level screening feature fusion pyramid network (CA-HSFPN) is adopted, which significantly reduces computational complexity while improving semantic representation. Furthermore, the Shape-IoU loss function is introduced to replace the traditional CIoU loss, achieving more accurate bounding box regression through geometric alignment that accounts for object shape. Experimental results on a custom peach dataset show that Peach-YOLO, with a compact model size of only 5.0 MB, achieves a real-time inference speed of 115.7 FPS, an mAP@0.5 of 82.2%, a precision of 78.9%, and a recall of 76.4%. Compared with the baseline YOLOv8n, the mAP, precision, and recall are improved by 3.0, 3.6, and 4.8 percentage points, respectively. Compared with current mainstream detection models, Peach-YOLO demonstrates superior performance in both accuracy and efficiency, providing a lightweight, high-precision, and real-time visual detection solution for automated fruit picking systems.

The degradation of organic pollutants in plants involves uptake and diffusion through the roots, trunk or leaves, transformation, accumulation and/or volatilization of soil- and aqueous-phase contaminants. 2,4,6-trinitrotoluene (TNT), the most widely used explosive, is toxic to a number of photosynthetic organisms, poses hazards to human health and pollutes the environment because it is recalcitrant to degradation. In this study, we investigated the nitroreductase gene from Citrobacter freundii in transgenic Arabidopsis thaliana plants. Transgenic plants showed a promising ability to tolerate, take up and detoxify TNT. Our results suggest that transgenic plants show higher potential for removing and transformation of TNT. The expression of nitroreductase in Arabidopsis could be useful for phytoremediation and could be explored for the effective cleanup of TNT-contaminated sites.

Ammonium (NH4+) plays key roles in plant growth, development, fruit quality, and yield. In plants, NH4+ uptake and transport are facilitated by NH4+ transporters (AMT). However, molecular mechanisms and physiological functions of type-II AMT (AMT2) transporters in fruit trees are still unclear, especially in peach. In this study, we cloned and characterized an AMT2 family gene from peach, PpeAMT3;4, and determined its function in yeast mutant. Expression analysis showed that PpeAMT3;4 was majorly expressed in peach roots and significantly decreased by NH4+ excess but had no response to NH4+ deficiency. Functional determination and 15nitrogen-labeled NH4+ uptake assay in yeast cells implied that PpeAMT3;4 was a typical high-affinity transporter, with a Km value of 86.3 μM, that can uptake external NH4+ in yeast cells. This study provides gene resources to uncover the biological function of AMT2 transporters and reveals molecular basis for NH4+ uptake and nitrogen (N) nutrition mechanisms in fruit trees.

KEY MESSAGE : The glyphosate resistance in Escherichia coli and Arabidopsis was due to d -amino acid oxidase expression. Transgenic glyphosate-resistant crops have a high percentage in the total area devoted to transgenic crops worldwide. D-amino acid oxidase (DAAO) can metabolize glyphosate by oxidative cleavage of the carbon–nitrogen bond on the carboxyl side and yield aminomethyl phosphonic acid and glyoxylate, which are less toxic to plants than glyphosate. To date, reports on the use of DAAO to enhance glyphosate resistance in plants are lacking. In this paper, we report synthesis, and codon usage optimization for plant expression, of the DAAO gene by successive polymerase chain reaction from Bradyrhizobium japonicum. To confirm the glyphosate resistance of the DAAO gene, the recombinant plasmid pYPX251 (GenBank Accession No: AY178046) harboring the wild-type DAAO gene was transformed into DH5α. The positive transformants grew well both on solid and in liquid M9 medium containing 200 mM glyphosate. The optimized DAAO gene was transformed into Arabidopsis and 9 days after application of 10 mM glyphosate, the 4-week-old wild-type plants all died; by contrast, transgenic plants could grow normally. The proline content and peroxidase activity showed that glyphosate could induce proline accumulation and produce reactive oxygen species.

Xanthine dehydrogenase (EC1.1.1.204; XDH) plays an important role in purine catabolism that catalyzes the oxidative hydroxylation of hypoxanthine to xanthine and of xanthine to uric acid. Long attributed to its role in recycling and remobilization of nitrogen, recently, XDH is implicated in plant stress responses and acclimation, such research efforts, however, have thus far been restricted to Arabidopsis XDH -knockdown/knockout studies. This study, using an ectopic overexpression approach, is expected to provide novel findings. In this study, a XDH gene from Vitis vinifera , named VvXDH , was synthesized and overexpressed in Arabidopsis , the transgenic Arabidopsis showed enhanced salt tolerance. The VvXDH gene was investigated and the results demonstrated the explicit role of VvXDH in conferring salt stress by increasing allantoin accumulation and activating ABA signaling pathway, enhancing ROS scavenging in transgenic Arabidopsis . In addition, the water loss and chlorophyll content loss were reduced in transgenic plants; the transgenic plants showed higher proline level and lower MDA content than that of wild-type Arabidopsis , respectively. In conclusion, the VvXDH gene has the potential to be applied in increasing allantoin accumulation and enhancing the tolerance to abiotic stresses in Arabidopsis and other plants.

Cancer development is closely associated with immunosuppressive tumor microenvironment (TME) that attenuates antitumor immune responses and promotes tumor cell immunologic escape. The sequential conversion of extracellular ATP into adenosine by two important cell-surface ectonucleosidases CD39 and CD73 play critical roles in reshaping an immunosuppressive TME. The accumulated extracellular adenosine mediates its regulatory functions by binding to one of four adenosine receptors (A1R, A2AR, A2BR and A3R). The A2AR elicits its profound immunosuppressive function via regulating cAMP signaling. The increasing evidence suggests that CD39, CD73 and A2AR could be used as novel therapeutic targets for manipulating the antitumor immunity. In recent years, monoclonal antibodies or small molecule inhibitors targeting the CD39/CD73/A2AR pathway have been investigated in clinical trials as single agents or in combination with anti-PD-1/PD-L1 therapies. In this review, we provide an updated summary about the pathophysiological function of the adenosinergic pathway in cancer development, metastasis and drug resistance. The targeting of one or more components of the adenosinergic pathway for cancer therapy and circumvention of immunotherapy resistance are also discussed. Emerging biomarkers that may be used to guide the selection of CD39/CD73/A2AR-targeting treatment strategies for individual cancer patients is also deliberated.

Non-negative integer-valued time series are usually encountered in practice, and a variety of integer-valued autoregressive processes based on various thinning operators are commonly used to model these count data with temporal dependence. In this paper, we consider a first-order integer-valued autoregressive process constructed by the negative binomial thinning operator with random coefficients, to address the problem of constant thinning parameters which might not always accurately represent real-world settings because of numerous external and internal causes. We estimate the model parameters of interest by the two-step conditional least squares method, obtain the asymptotic behaviors of the estimators, and furthermore devise a technique to test the constancy of the thinning parameters, which is essential for determining whether or not the proposed model should consider the parameters’ randomness. The effectiveness and dependability of the suggested approach are illustrated by a series of thorough simulation studies. Finally, two real-world data analysis examples reveal that the suggested approach is very useful and flexible for applications.

Existing approaches for admissibility analysis of T-S fuzzy descriptor time delay systems fail to balance conservatism reduction and computational complexity. This paper proposes a low-conservatism analysis and stabilization method based on the symmetric Lyapunov–Krasovskii (L-K) functional. By exploiting the boundedness of membership function derivatives, and combining Jensen’s integral inequality with auxiliary slack matrices to achieve tight bounding of nonlinear terms, we derive an admissibility criterion for open-loop systems with significantly reduced conservatism. A well-suited L-K functional is constructed targeting the structural characteristics of fuzzy singular matrices Eξ, a state feedback controller is designed via the parallel distributed compensation (PDC) strategy, and solvable sufficient conditions for the admissibility of closed-loop systems are established. Numerical examples demonstrate that the maximum allowable delay upper bound obtained by the proposed method outperforms that of existing state-of-the-art approaches while balancing conservatism and computation cost and verifying the superiority of the proposed method.