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Rapid on-site genotyping of the ovine prolific FecBB mutation using a CRISPR/Cas12a-based detection system
Tingjie Wu, Jiayuan Sun, Lijin Lu, Chen Wang, Shiwei Zhou, Yulin Chen, Xinjie Wang, Xiaolong Wang
2026, 25 (4): 1597-1605.   DOI: 10.1016/j.jia.2024.05.013
Abstract134)      PDF in ScienceDirect      

BMPR1B is a pivotal gene that influences reproductive performance in sheep.  The sheep populations that carry the FecBB mutation within this gene exhibit significantly higher lambing rates compared to wild-type populations.  Therefore, screening for individuals carrying the FecBB 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 FecBB 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 FecBB genotyping.  It holds promise in enhancing efficiency in livestock breeding programmes for any single nucleotide mutations.  

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Sheep with partial RXFP2 knockout exhibit normal horn phenotype but unilateral cryptorchidism
Yawei Gao, Siyuan Xi, Bei Cai, Tingjie Wu, Qian Wang, Peter Kalds, Shuhong Huang, Yuhui Wang, Saizheng Han, Menghao Pan, Chong Yang, Qifang Kou, Baohua Ma, Xiaolong Wang, Shiwei Zhou, Yulin Chen
2025, 24 (9): 3698-3702.   DOI: 10.1016/j.jia.2023.11.045
Abstract290)      PDF in ScienceDirect      
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Anaerobic soil disinfestation rather than Bacillus velezensis Y6 inoculant suppresses tomato bacterial wilt by improving soil quality and manipulating bacterial communities
Taowen Pan, Yulin Chen, Sicong Li, Lei Wang, Joji Muramoto, Carol Shennan, Jihui Tian, Kunzheng Cai
2025, 24 (2): 754-768.   DOI: 10.1016/j.jia.2024.06.019
Abstract215)      PDF in ScienceDirect      
Continuous cropping leads to high incidence of soilborne diseases such as bacterial wilt caused by Ralstonia solanacearum, which poses a risk to agricultural production.  Anaerobic soil disinfestation (ASD) and plant growth-promoting rhizobacteria (PGPR) are considered environmentally friendly methods to control bacterial wilt.  However, the underlying mechanism of the improvement of soil health and the inhibition of bacterial wilt after ASD treatment and PGPR inoculation needs further exploration.  This study evaluated the effect of ASD treatment on soil improvement at pre-planting of tomato, and the effect of ASD treatment combined with the application of Bacillus velezensis Y6 (BV) on soil quality, Rsolanacearum abundance, and bacterial communities at 90 days before harvesting of tomato.  The results showed that ASD treatment reduced Rsolanacearum abundance in soil by 17.6% at pre-planting and 18.7% at 90 days before harvesting, but BV inoculation did not influence R. solanacearum abundance.  ASD and ASD+BV treatments effectively reduced the occurrence of bacterial wilt, improved soil nutrient status and increased soil microbial activity at 90 days before harvesting.  Principal co-ordinate analysis showed that the soil bacterial community was significantly influenced by ASD treatment both at pre-planting and at 90 days before harvesting.  Further investigation found that ASD contributed to the enrichment of beneficial flora (Bacillus and Streptomyces).  Moreover, pH was an important environmental factor affecting the abundance of Rsolanacearum in soil.  Co-occurrence network analysis showed that ASD treatment significantly increased network connection of bacterial communities and the proportion of beneficial microorganisms (Proteobacteria and Firmicutes), leading to complex soil bacterial co-occurrence networks both at pre-planting and at 90 days before harvesting.  Collectively, these results indicate that ASD treatment, but not microbial inoculation can enhance tomato plant resistance to bacterial wilt by improving soil quality and modulating the soil bacterial community.
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