Ascochyta blight (AB) is a devastating fungal disease of chickpeas that has spread to nearly all of the chickpea cultivating regions of the world. The rapid diagnosis of Ascochyta rabiei L. (A. rabiei), the pathogen that causes AB, plays an important role in A. rabiei epidemic tracking and AB management. In this study, a group of loop-mediated isothermal amplification (LAMP) primers was designed to detect A. rabiei in chickpea plants and seeds via a LAMP method and a conventional PCR method based on an internal transcribed spacer (ITS) sequence analysis of A. rabiei. Compared with the conventional PCR method, the LAMP method not only exhibited greater sensitivity and specificity in the detection of A. rabiei but also used simpler equipment and required less operational time. The minimum detectable concentration of the A. rabiei genomic DNA solution with the LAMP method was 6.01 × 10⁻⁶ng/μl, which was 100 times lower than that of the conventional PCR method with the same outer primers. The greatest advantage of the LAMP method is that results can be observed via the visualization of color changes in SYBR Green I dye with the naked eye, and it does not require expensive instruments, also with less time consumption.

Background: Neonicotinoids are widely applied in the control of the destructive agricultural pest Bemisia tabaci, and resistance against these chemicals has become a common, severe problem in the control of whiteflies. To investigate the molecular mechanism underlying resistance against nenonicotinoids in whiteflies, RNA-seq technology was applied, and the variation in the transcriptomic profiles of susceptible whiteflies and whiteflies selected by imidacloprid, acetamiprid and thiamethoxam treatment was characterized. Results: A total of 90.86 GB of clean sequence data were obtained from the 4 transcriptomes. Among the 16,069 assembled genes, 584, 110 and 147 genes were upregulated in the imidacloprid-selected strain (IMI), acetamiprid-selected strain (ACE), and thiamethoxam (THI)-selected strain, respectively, relative to the susceptible strain. Detoxification-related genes including P450s, cuticle protein genes, GSTs, UGTs and molecular chaperone HSP70s were overexpressed in the selected resistant strains, especially in the IMI strain. Five genes were downregulated in all three selected resistant strains, including 2 UDP-glucuronosyltransferase 2B18-like genes (LOC 109030370 and LOC 109032577). Conclusions: Ten generations of selection with the three neonicotinoids induced different resistance levels and gene expression profiles, mainly involving cuticle protein and P450 genes, in the three selected resistant whitefly strains. The results provide a reference for research on resistance and cross-resistance against neonicotinoids in B. tabaci.

Verticillium wilt is one of the most crucial diseases caused by Verticillium dahliae that threatens the cotton industry. Statistical results showed that the return of cotton plants infected with V. dahliae to the field might be an essential cause of the continuous aggravation of cotton Verticillium wilt. The correlation among the cotton plants infected with V. dahliae returning to the field, the occurrence of Verticillium wilt, and the number of microsclerotia in rhizosphere soil need further investigation. A potted experiment was carried out to explore the effects of the direct return of cotton plants infected with Verticillium dahliae to the field on the subsequent growth and Verticillium wilt occurrence in cotton. As a risk response plan, we investigated the feasibility of returning dung-sand (i.e., insect excreta) to the field, the dung-sand was from the larvae of Protaetia brevitarsis (Coleoptera: Cetoniidea) that were fed with the V. dahliae–infected cotton plants. The results demonstrated that the return of the entire cotton plants to the field presented a promotional effect on the growth and development of cotton, whereas the return of a single root stubble or cotton stalks had an inhibitive effect. The return of cotton stalks and root stubble infected with V. dahliae increased the risk and degree of Verticillium wilt occurrence. The disease index of Verticillium wilt occurrence in cotton was positively correlated with the number of microsclerotia in the rhizosphere soil. The disease index increased by 20.00%, and the number of soil microsclerotia increased by 8.37 fold in the treatment of returning root stubble infected with V. dahliae to the field. No Verticillium wilt microsclerotia were detected in the feed prepared from cotton stalks and root stubble fermented for more than 5 days or in the transformed dung-sand. There was no risk of inoculation with Verticillium wilt microsclerotia when the dung-sand was returned to the field. The indirect return of cotton plants infected with V. dahliae to the field by microorganism–insect systems is worthy of further exploration plan of the green prevention and control for Verticillium wilt and the sustainable development of the cotton industry.