Heterodera schachtii is a well-known cyst nematode that causes serious economic losses in sugar beet production every year. Rapid and visual detection of H. schachtii is essential for more effective prevention and control. In this study, a species-specific recombinase polymerase amplification (RPA) primer was designed from a specific H. schachtii sequence-characterized amplified region (SCAR) marker. A band was obtained in reactions with DNA from H. schachtii, but absent from nontarget cyst nematodes. The RPA results could be observed by the naked eye, using a lateral flow dipstick (LFD). Moreover, we combined CRISPR technology with RPA to identify positive samples by fluorescence detection. Sensitivity analysis indicated that 10⁻⁴ single cysts and single females, 4⁻³ single second-stage juveniles, and a 0.001 ng genomic DNA template could be detected. The sensitivity of the RPA method for H. schachtii detection is not only higher than that of PCR and qPCR, but can also provide results in <1 h. Consequently, the RPA assay is a practical and useful diagnostic tool for early diagnosis of plant tissues infested by H. schachtii. Sugar beet nematodes were successfully detected in seven of 15 field sugar beet root samples using the RPA assay. These results were consistent with those achieved by conventional PCR, indicating 100% accuracy of the RPA assay in field samples. The RPA assay developed in the present study has the potential for use in the direct detection of H. schachtii infestation in the field.

Sugar beet cyst nematode (SBCN, Heterodera schachtii) is an important nematode that causes significant yield losses of 25–50% or more in most areas of sugar beet production worldwide. Rapid and accurate identification of this species is essential to support decisions on pest management. However, the difference between H. schachtii and other Heterodera spp. based on morphology is a challenging task. In the present study, a SCAR-PCR assay was developed to identify and differentiate H. schachtii in infected root and soil samples. H. schachtii-species-specific SCAR-PCR primers OPA06-HsF and OPA06-HsR were designed from the randomly amplified polymorphic DNA (RAPD) marker amplified with random primer OPA06. The developed primers specifically amplify a 922-bp fragment from the target populations but did not amplify DNA from non-target cyst nematodes including Heterodera, Globodera, Cactodera, and other related species tested in this study. The sensitivity detection indicated that 5 × 10⁻⁴ of a single cyst, 1/320 of a single second-stage juvenile (J2), or 10 pg of genomic DNA could be detected. The assay accurately identifies the different stages of H. schachtii in sugar beet and oilseed rape roots as well as a single J2 in 10 g of soil. Finally, the SCAR-PCR assay detected H. schachtii in seven samples out of the fifteen field samples. The assay will not only be useful for differentiating H. schachtii from mixed populations of Heterodera spp. but also for effective detection of the species directly from infested samples. The assay also requires no expertise in the taxonomy and morphology of the species but serves to improve the diagnosis of H. schachtii in infested fields.

β-D-xylosidase (BXL) is a key enzyme involved in xylan degradation and plays crucial roles in plant development and stress responses. However, its functional roles in potato (Solanum tuberosum) remain poorly understood. In this study, we performed a genome-wide identification of the StBXL gene family and identified eight StBXL genes distributed across five chromosomes. A phylogenetic analysis classified these genes into four groups. Conserved motif and domain analyses indicated functional conservation among StBXL proteins. Analyses of cis-acting elements in the promoters and expression profiles of StBXL genes in various plant tissues under different stress treatments revealed their spatiotemporal expression patterns, as well as potential roles in phytohormone signaling and stress responses. Alkali stress significantly inhibited the expression of StBXL4 and StBXL5. The overexpression of StBXL4 enhanced the sensitivity of potato seedlings to alkali stress, whereas the overexpression of StBXL5 showed no significant phenotypic differences under the same conditions. These results suggest that StBXL4 acts as a negative regulator of the alkali stress response in potato. This study fills the research gap regarding the potato StBXL gene family and provides valuable insights for the molecular breeding of alkali-tolerant potato varieties.

In rice (Oryza sativa L.), the short-chain dehydrogenase protein OsABA2 plays a crucial role in regulating abscisic acid (ABA) biosynthesis. However, little is known about the other proteins that interact with OsABA2 to regulate ABA biosynthesis. Using yeast two-hybrid screening, we identified a novel OsABA2 interacting protein OsPP2C55, which contains a serine/threonine phosphatase (family 2C) catalytic domain. The yeast two-hybrid (Y2H) assay and firefly luciferase complementary imaging (LCI) assay confirmed these interactions. Subsequent studies revealed that saline–alkaline stress significantly downregulated OsPP2C55 gene expression. Meanwhile, we constructed ospp2c55 CRISPR gene knockout (ospp2c55-KO) plants using Agrobacterium genetic transformation. Compared with wild-type plants, ospp2c55-KO plants under saline–alkaline stress exhibited significantly elevated OsABA2 protein levels, leading to substantial increases in ABA content. In addition, ospp2c55-KO plants demonstrated heightened sensitivity to ABA during seed germination. Moreover, ospp2c55-KO plants improved the survival rate and stress-related indices of rice seedlings under saline–alkaline stress, and upregulated the expression of genes related to adversity stress (OsNCED1, OsNCED3, OsABA2, OsSODCc2, and OsCatB). We found that OsPP2C55 plays a negative regulatory role in ABA biosynthesis and saline–alkaline stress tolerance in rice.