BMPR1B is a pivotal gene that influences reproductive performance in sheep.  The sheep populations that carry the FecB^B mutation within this gene exhibit significantly higher lambing rates compared to wild-type populations.  Therefore, screening for individuals carrying the FecB^B mutation is crucial for effective sheep breeding programs.  This study aims to establish a rapid, precise, and visualised on-site detection method for genotyping the prolific FecB^B mutation in sheep.  We combined the CRISPR/Cas12a system with the recombinase-polymerase amplification (RPA) technique.  We introduced an additional nucleotide mismatch on the amplification primers to form a Cas12a-recognised protospacer adjacent motif (PAM) sequence.  In addition, mismatches were introduced in CRISPR-derived RNA (crRNA) to enable naked-eye differentiation of the assay results.  Subsequently, we validated the accuracy of the method by examining additional blood samples from 56 sheep representing four breeds.  The results of using our developed system were highly consistent with the Sanger sequencing.  Overall, the CRISPR/Cas12a-based detection provides a rapid and more versatitle method for FecB^B genotyping.  It holds promise in enhancing efficiency in livestock breeding programmes for any single nucleotide mutations.  

The postharvest senescence phase of table grapes comprises a series of biological processes.  MicroRNAs (miRNAs) regulate downstream genes at the post-transcriptional level; however, whether miRNAs are involved in postharvest grape senescence remains unclear.  We used small RNA sequencing to identify postharvest-related miRNAs in ‘Red Globe’ (Vitis vinifera) grapes harvested after 0, 30, and 60 d of storage at 4°C (RG0, RG30, RG60).  In total, 42 known and 219 novel miRNA candidates were obtained.  During fruit senescence, the expression of PC-3p-3343_1921, miR2950, miR395k, miR2111, miR159c, miR169q, PC-5p-1112_4500, and miR167b changed significantly (P<0.05).  Degradation sequencing identified 218 targets associated with cell wall organization, tricarboxylic acid (TCA) cycling, pathogen defense, carbon metabolism, hormone signaling, the anthocyanin metabolism pathway, and energy regulation, of which ARF6, GRF3, TCP2, CP1, MYBA2, and WRKY72 were closely related to fruit senescence.  We also verified that VIT_00s2146g00010, VIT_02s0012g01750, and VIT_03s0038g00160 with unknown functions are cleaved by senescence-related PC-5p-1112_4500 via the dual luciferase assay, and the transient transformation of grape berries showed that they regulate berry senescence.  These results deepen our understanding of the role of miRNAs in regulating grape berry senescence and prolonging the shelf life of horticultural products.  Based on these results, we propose a new theoretical strategy for delaying the postharvest senescence of horticultural products by regulating the expression of key miRNAs (e.g., PC-5p-1112_4500), thereby extending their shelf life.

Heat stress (HS) can induce the disruption of small intestinal integrity in broilers. Here we examined the efficacies and possible mechanisms of zinc proteinate with moderate chelation strength (Zn-Prot M) compared to ZnSO₄.7H₂O (ZnSO₄) in alleviating HS-induced disruption of small intestinal integrity in broilers. The experiment included 7 treatments. Twenty two-d-old birds were randomly allotted into 1 of 5 treatments under high temperature (HT, 9:00-17:00: 34±1°C, 8 h d^-1; 17:00–9:00: 28±1°C) with 1 control (HT-CON) plus 2 [dietary Zn sources, Zn-Prot M and ZnSO₄] × 2 [added Zn levels, 30 and 60 mg kg^-1] factorial arrangement, and 1 of 2 treatments under normal temperature (NT) with the control (NT-CON) and pair-feeding matched to HT-CON (NT-PF). The results showed that HS dramatically reduced (P<0.05) small intestinal villus height (VH), VH/crypt depth (CD) and villus surface area (VSA), the amount of proliferating cell nuclear antigen (PCNA) positive cells, mRNA or protein expression levels of phosphatidylinositol 3-kinase (PI3K), serine threonine kinase (AKT), extracellular regulated protein kinase (ERK), protein kinase C (PKC), G protein-coupled receptor 39 (GPR39), phosphorylated PI3K (p-PI3K)/PI3K, p-AKT/AKT, p-ERK/ERK and phospholipase C (PLC) β1/2, and PLC activity. HS also remarkably increased (P<0.05) small intestinal CD and mRNA or protein levels of P38 mitogen activated protein kinase (P38 MAPK), C-jun N-terminal kinase (JNK)1/2 and p-JNK/JNK in the jejunum of heat stressed (HS) broilers. However, dietary supplementation with Zn, especially organic Zn as Zn-Prot M at 60 mg kg^-1, significantly increased (P<0.05) small intestinal VH, VH/CD and VSA, as well as the amount of PCNA positive cells, PLC activity, mRNA expression levels of PI3K, AKT, GPR39 and PLC β1, protein expression levels of p-PI3K/PI3K, p-AKT/AKT, p-ERK/ERK, GPR39 and PLC β1, and decreased (P<0.05) small intestinal CD, JNK2 mRNA expression level and p-JNK/JNK protein expression level in the jejunum of HS broilers. These results suggest that dietary supplemental Zn, especially 60 mg Zn kg^-1 as Zn-Prot M, can effectively alleviate HS-induced small intestinal injury possibly by promoting cellular proliferation via the GPR39/PLC β1-mediated PI3K/AKT or MAPK pathways in the jejunum of HS broilers.

The fungal pathogen Ustilaginoidea virens, which causes the devastating rice false smut disease, relies on scavenging host nutrients for infection, yet the identity and exploitation mechanisms of these nutrients remain unclear. This study reveals a previously unrecognized virulence strategy in which U. virens hijacks host-derived trehalose, a disaccharide linked to plant stress responses, by enhancing its trehalose catabolic capacity. Central to this process are two functionally distinct trehalases, UvNTH (neutral trehalase) and UvATH (acid trehalase), which exhibit a clear division of labor. UvNTH drives extracellular trehalose utilization, vegetative growth, and asexual sporulation, while UvATH specifically regulated aerial hyphal development. Strikingly, the ΔUvNTH mutant colonizes host tissues normally but completely fails to produce mature false smut balls, uncoupling colonization from symptom formation and identifying UvNTH as a dedicated virulence switch essential for pathogenicity. In addition, UvNTH plays a critical regulatory role in the germination process of chlamydospores, the primary inoculum of the pathogen. Collectively, these findings demonstrate that UvNTH functions not only as a metabolic enzyme but also as a dual-functional regulator that coordinates virulence and the propagation of primary infection sources, thereby providing an ideal target for the sustainable management of rice false smut.