A high-throughput phenotyping procedure for evaluation of antixenosis against common cutworm at early seedling stage in soybean

Background: Common cutworm (CCW; Spodoptera litura Fabricius) is a major leaf-feeding pest of soybean in Asia. The previous methods of measuring antixenosis against CCW using adult plant under field or net-room conditions were time-consuming, labor-intensive and precision-inferior. To solve the problems, this study aimed at (i) establishing a high-throughput phenotyping method for evaluating antixenosis against CCW at early seedling stage, (ii) using the procedure to evaluate the antixenosis of an insect-resistant versus-susceptible germplasm population (IRSGP), (iii) validating the proposed method through comparing the results with the historical phenotypic data and phenotyping- genotyping consistency data using PAV (presence/absence variation) markers linked with the identified loci CCW-1 and CCW-2, (iv) and evaluating the efficiency of the novel method through comparisons to the previous methods. Results: A dynamic and efficient evaluation procedure characterized with using V1 stage soybean seedlings infested with third-instar larvae in a micro-net-room in greenhouse with damaged leaf percentage (DLP) as indicator was established and designated V1TMD method. The middle term testing stage is the best dates for identifying resistant and susceptible accessions. The results from the V1TMD method were relatively stable, precise and accurate in comparison with the previous method with the detected most resistant and susceptible accessions consistent to the previous results. The DLP values differentiated obviously to coincide with the resistant and susceptible alleles of the PAV markers Gm07PAV0595 and Gm07PAV0389 tightly linked to the two resistance-related loci, CCW-1 and CCW-2, respectively, in IRSGP. Thus V1TMD is a high-throughput phenotyping method with its estimated efficiency 12 times and period shortening 4 times of those of the previous method. Conclusion: A dynamic and efficient V1TMD method for testing antixenosis against CCW was established, with highly resistant and highly susceptible accessions as standard checks and DLP as indicator. The method is remarkably quick, highly reproducible, and capable of testing large samples, therefore, is a high-throughput phenotyping method.