Arthropods serve essential roles in crop production as pollinators, predators, and pests.  Understanding arthropod biodiversity is crucial for assessing agroecosystem health, functions, and services.  Traditional survey methods are labor-intensive, costly, and rely on diminishing taxonomic expertise, limiting their agricultural applications.  Environmental DNA (eDNA) metabarcoding of diverse samples provides comprehensive species composition data through efficient and non-invasive sampling.  However, this method remains underutilized in rice field studies.  This research examined four sample substrates - rice plant cleaning fluid (RPCF), rice pollen, soil, and water - using various barcoding primers to identify optimal substrates for monitoring rice paddy arthropod diversity.  The method was implemented in Bt rice and non-Bt rice fields to evaluate its biomonitoring potential.  Results indicate that the COI primer (mlCOIintF/jgHCO2198R) identified the highest number of rice field arthropod species.  The eDNA collected from RPCF detected 15% more arthropod species compared to vacuum sampling of whole arthropods.  Rice pollen collection during the heading stage also revealed considerable arthropod diversity.  Alpha diversity and taxonomic composition remained consistent between Bt and non-Bt rice fields, aligning with traditional survey findings.  These results suggest that eDNA metabarcoding of plant cleaning fluid offers an effective approach for monitoring agricultural arthropod communities, contributing to agricultural production optimization.

Peanuts (Arachis hypogaea) are important sources of vegetable oil, protein, and forage.  The genus Arachis comprises nine intrageneric taxonomic sections encompassing 84 species.  Most Arachis species are wild plants that serve widely as forage and turfgrass.  Furthermore, wild Arachis species provide valuable gene resources for broadening the genetic diversity of cultivated peanuts.  To date, several key genes have been identified through the use of recombinant inbred lines derived from interspecific crosses within Arachis.  Despite this progress, the application of genetic engineering to enhance peanut traits remains limited.  This limitation arises primarily from the absence of a robust and reliable genetic transformation protocol for Arachis species.  Nevertheless, evidence indicates that successful genetic transformation of Arachis plants was first reported approximately 30 years ago.  Thus, a notable discrepancy exists between early reports of transformation success and the ongoing challenges in stably transferring candidate genes into Arachis genotypes.  This review summarizes existing methods for regeneration and genetic transformation in Arachis, aiming to advance understanding of transgenic technologies applicable to this genus.

NADC34-like porcine reproductive and respiratory syndrome virus (PRRSV), which first appeared in China in 2017, is currently one of the main epidemic strains in China.  In this study, we found that a new variant of NADC34-like PRRSV evolved, named the L1A variant.  The phylogenetics, epidemic status, and pathogenicity of the LA variants were subsequently comprehensively evaluated.  Based on the results of the ORF5 phylogenetic analysis, the L1A variants were classified as NADC34-like PPRSV.  All the strains had the same discontinuous 131-aa deletion in the NSP2 region (similar to that in the NADC30).  Recombination analysis revealed that the L1A variants were recombinant viruses that contained an NADC30-like PRRSV skeleton, a nonstructural protein-encoding gene region obtained in part from JXA1-like PRRSV and a ORF2-ORF6 gene region partly obtained from NADC34-like PRRSV and that exhibited similar recombination patterns.  We successfully isolated the L1A variant TZJ2756 from PAMs and Marc-145 cells.  In animal experiments, TZJ2756 exhibited moderate pathogenicity in piglets, causing obvious clinical symptoms, namely, persistent fever, significantly reduced body weight, interstitial edema and severe interstitial pneumonia in the lungs, and prolonged high-load viremia.  L1A variants have been detected in at least 12 provinces in China and share many similar epidemiological characteristics with the American L1C variant.  This research will enhance our understanding of the prevalence of L1A variants and furnish valuable data for the ongoing monitoring of NADC34-like PRRSV in China.

Red fruit peel is one of pear’s most valuable economic traits and is mainly determined by anthocyanins.  Many pear cultivars with a red peel originated from bud sports; however, little is known about the genetic mechanisms underlying this trait.  We have previously identified a mutant PpBBX24 containing a 14-nucleotide deletion in the coding region (Ppbbx24-del) as the only known variant associated with the red coloration of the mutant ‘Red Zaosu’ pear (Pyrus pyrifolia White Pear Group).  Herein, we analyzed the role of the mutant gene in red coloration and its mechanism of action.  The results showed that light promoted red peel coloration in the ‘Red Zaosu’ pear, and Ppbbx24-del positively affected light-induced anthocyanin biosynthesis, while normal PpBBX24 had the opposite effects.  Transient and stable transformation experiments confirmed that Ppbbx24-del could promote anthocyanin accumulation in pear fruit peels, calli, and tobacco flowers.  Due to the loss of nuclear localization sequence (NLS) and viral protein (VP) domains, Ppbbx24-del co-localized in the nucleus and cytoplasm, whereas PpBBX24 localized only in the nucleus.  Real-time PCR and transcriptome analyses indicated that PpMYB10 and PpHY5 are highly expressed in the ‘Red Zaosu’ pear.  In yeast one-hybrid and dual-luciferase assays, Ppbbx24-del and PpHY5 independently promoted the expression of PpCHS, PpCHI, and PpMYB10 by binding to their promoters; however, PpBBX24 did not affect the expression of these genes.  Additionally, we found that Ppbbx24-del and PpHY5 had additive effects on the expression of PpCHS, PpCHI, and PpMYB10, as they promote the expression of anthocyanin synthesis genes separately.  The co-expression of PpBBX24 and PpHY5 inhibited the activation of downstream genes by PpHY5, which was attributed to the interaction between the two loci.  In conclusion, our results clarify the molecular mechanism by which mutant Ppbbx24-del and PpBBX24 exert opposite effects in regulating anthocyanin accumulation in pear.  These findings lay an important theoretical foundation for using Ppbbx24-del to create red pear cultivars.

Anthracnose is a devastating disease caused by Colletotrichum that significantly affects the yield and economic value of the tea plant (Camellia sinensis).  However, few studies have addressed the genetic mechanism of anthracnose resistance (AR).  This study investigated the QTL associated with AR in a ‘Longjing 43’בBaijiguan’ (LJ43×BJG) population.  The field surveys conducted in this study led to the identification of several QTLs for AR on the linkage map.  One major QTL (qAR-12.4) accounted for 12% of the phenotypic variance explained over two years.  The BSA-seq results also revealed two genomic regions, qARChr1 on chromosome 1 and qARChr13 on chromosome 13, which showed strong correlations with AR.  Time-course RNA-seq was performed on LJ43 and BJG inoculated with anthracnose at 0, 24, and 48 hours to screen for candidate genes.  The results showed the gradual post-inoculation expression of a nuclear-localized ERF transcription factor (CsERF105) within the qARChr1 locus in BJG but not in LJ43.  The AR of BJG was significantly reduced after feeding with CsERF105-specific antisense oligonucleotides, suggesting that CsERF105 may be a positive regulator.  The findings of this study add to our general knowledge of the genetic factors involved in the tea plant’s AR and potential breeding targets.

Bud dormancy is a crucial adaptation for perennial fruit plants, enabling them to withstand unfavorable growth conditions. This adaptive strategy plays a significant role in the survival and reproduction of these plants, yet its molecular basis is not fully understood. In the current study, two transcription factors in grapes, EARLY BUD BREAK (VvEBB) and SHORT VEGETATIVE PHASE 4 (VvSVP4), were identified and examined. The findings demonstrated that, following heterologous transformation in poplar, VvSVP4 functions as a negative regulator, whereas VvEBB acts as a positive regulator in the process of bud-break. Transcriptome analysis showed that plant hormone signaling pathways, specifically those involving abscisic acid (ABA), indole acetic acid (IAA), and cytokinin (CK), were significantly enriched in plants overexpressing VvSVP4 (VvSVP4oe) and VvEBB (VvEBBoe), compared to control plants. Additionally, changes in the endogenous levels of ABA, IAA, and CK were found to be positively correlated with the transcriptome data. During the endodormancy phase, VvSVP4 directly and positively influenced the expression of the ABA receptor gene VvPYL9, thereby maintaining the state of bud dormancy. Conversely, during ecodormancy, the VvEBB gene was rapidly upregulated and negatively impacted the expression of the sucrose nonfermenting 1-related protein kinase subfamily 2 gene (VvSAPK2), facilitating the release from dormancy. In summary, this study offers a comprehensive explanation of the roles of VvSVP4 and VvEBB genes in dormancy and bud break, integrating insights into cell cycle regulation and multiple hormones signaling pathways.

Starch serves as a critical storage component, significantly influencing the grain yield and quality of maize (Zea mays L.).  Understanding the genetic basis of natural variation in kernel starch content (SC) is essential for maize breeding to meet future demands.  A genome-wide association study (GWAS) identified 84 and 96 loci associated with kernel SC across two years, overlapping with 185 candidate genes.  The candidate gene ZmMYB71, encoding a MYB-related transcription factor, demonstrated the highest co-expression frequency with starch synthesis genes.  Analysis revealed that ZmMYB71 functions as a nuclear located transcription repressor, and mutants exhibited increased kernel SC by over 2.32%, with minimal impact on amylose content or 100-grain weight.  Sh1, Sh2, and GBSSI exhibited up-regulation in mutants by 1.56-, 1.45- and 1.32-fold, respectively, aligning with RNA sequencing results; their promoter activities appear directly repressed by ZmMYB71 through the GATATC and TTAGGG motifs.  Additionally, the ZmMYB71 elite haplotype Hap1 occurred in over 55% of the high-starch maize sub-populations Iowa Stiff Stalk Synthetic (BSSS) and Partner B (PB), but only in 7.14% of the low-starch sub-population Partner A (PA).  Analysis of Hap1 haplotype frequencies across breeding stages revealed a significant increase to 40.28% in inbred groups released after 2010, compared to 28.57 and 27.94% in 1980 and 1990, and 2000, respectively.  These findings enhance understanding of natural variation in maize kernel SC and establish ZmMYB71 as a negative regulator with potential applications in SC improvement.

In the central spikelet, floret primordia proximal to rachis typically develop into fertile florets, while those distal ones tend to abort.  However, the mechanism driving these divergent outcomes remain unclear.  To elucidate this mechanism, a two-year field experiment was conducted, analyzing the differences between fertile (floret 1) and abortive florets (floret 5) at phenotypic, physiological, cellular, and transcriptional levels.  Our results showed that floret 1 (F1) developed earlier than floret 5 (F5), evidenced as distinct floret and anther cell morphology.  This developmental difference was associated with distinct metabolic strategies: F1 prioritizes starch biosynthesis by enhancing photosynthetic efficiency and reducing thermal dissipation, while F5 diverts carbon to trehalose metabolism under resource constraints.  Transcriptomic profiling revealed 17,711 differentially expressed genes, predominantly enriched in carbohydrate metabolism.  Key starch-related genes (WAXY, SS, GBE1) were upregulated in F1, while trehalose synthesis genes (TPS.5, TPS.6, TPP.7) dominated in F5, reflecting a metabolic trade-off between growth and survival. Hormonal profiling revealed elevated indole-3-acetic acid (IAA) levels in F1, driven by upregulated genes encoding enzymes of TDC and YUCCA, whereas abscisic acid (ABA) level increased in F5, mediated by NCED.  Specially, tissue-specific regulation in fertile floret was clarified: lemma tissues enriched in photosynthesis genes supplied localized carbon, anthers relied on sucrose cleavage genes to sustain pollen development, and ovary activated lipid biosynthesis genes for embryogenesis.  Collectively, we propose a model where IAA-starch synergy in proximal florets establishes a competitive metabolic sink, whereas ABA-T6P interplay in distal florets enforces developmental arrest, thereby optimizing resource allocation and reproductive success.  The proposed regulatory framework provides novel views for improving floret fertility through carbohydrate partitioning and hormone signal.

During wheat grain filling, exogenous nitrogen supply can enhance grain yield and protein accumulation by delaying senescence and increasing nitrogen reserves. However, the underlying mechanisms remain unclear. The efficacy of canopy nitrogen spraying at 15 days after anthesis (AS) was first evaluated in a pot experiment, and the associated regulatory mechanisms were further investigated in a field trial under water-saving cultivation conditions. The pot experiment demonstrated that AS treatment increased grain weight, yield, and nitrogen accumulation by improving both pre-anthesis nitrogen remobilization and post-anthesis nitrogen assimilation. Canopy-derived nitrogen began accumulating significantly in grains at 12 h after spraying, accounting for 32.52% of the increase in grain nitrogen accumulation. The field experiment further validated that AS treatment increased grain filling rate and nitrogen accumulation rate during fast and slow growth stages, significantly increasing grain yield by 5.21% and protein content by 7.50% compared to spraying equal amounts of deionized water (CK). AS treatment upregulated key enzymes in the C₄ pathway—including phosphoenolpyruvate carboxylase (PEPC), NADP-malate dehydrogenase (NADP-MDH), NADP-malic enzyme (NADP-ME), pyruvate phosphate dikinase (PPDK)—and increased malate levels in glumes, lemmas, and paleae. These responses suggested that AS treatment facilitated the tricarboxylic acid (TCA) cycleand the Calvin cycle, providing reaction substrates for protein and starch biosynthesis. Additionally, AS treatment promoted grain nitrogen metabolism, facilitating protein accumulation. This study presents a viable strategy to mitigate post-anthesis drought stress and improve wheat productivity and grain quality in regions with similar agroclimatic conditions.

Brucellosis, caused primarily by Brucella abortus, Brucella melitensis, and Brucella suis, remains a critical global public health challenge, particularly in regions where these pathogens persist in livestock and wildlife reservoirs. Despite decades of control measures-including vaccination, test-and-removal programs, and biosecurity protocols-persistent human and animal cases highlight the limitations of existing diagnostic and intervention strategies. CRISPR-based diagnostics have emerged as a transformative tool, offering rapid, ultrasensitive, and field-deployable pathogen detection. Here, we present BOVDS (Brucella melitensis/abortus/suis-other pathogens-vaccine detection and differentiation system), an innovative CRISPR/Cas13a-based platform that integrates ultrahigh sensitivity (10 copies/µL), screening for 10 major abortifacient pathogens, and precise strain differentiation-overcoming key challenges in Brucella diagnostics. By incorporating mismatched spacer designs, BOVDS achieves robust discrimination between B. melitensis, B. abortus, and B. suis despite their high genomic conservation. Additionally, the platform enables differentiation between vaccine and wild-type strains, addressing critical gaps in vaccination monitoring and epidemiological surveillance. Uniting laboratory-level accuracy with on-farm practicality, BOVDS facilitates real-time outbreak management, targeted culling, and environmental decontamination, advancing One Health initiatives toward sustainable brucellosis prevention and control. This system sets a new benchmark for next-generation zoonotic disease diagnostics, with broad applicability in global public and veterinary health.