This project consists of two research components:

1. Development of Molecular Markers for Wilt Resistance in Luffa
   • During the spring cropping season this year, a root-cut inoculation assay was conducted on 20 F2:3 lines derived from the previously evaluated cross 4-5-1 × 157-5A. Among these, six lines that had also been inoculated in the autumn cropping season of 2025 (Year 114 of the ROC calendar) were included as reference families: two resistant, two susceptible, and two intermediate families. The resistant reference families consistently exhibited resistance in the inoculation assays conducted in both cropping seasons. Although not all plants in the susceptible reference families exhibited disease symptoms, their disease incidence remained higher than that of the intermediate reference families. Together with the F2:3 families evaluated in the autumn cropping season of 2025, a total of 45 F2:3 families have been evaluated. Since seeds for subsequent F2:3 testing were successfully obtained from only 45 F2 plants in the previous generation, bulked segregant analysis (BSA) is planned as an alternative approach to identify major-effect genes associated with disease resistance. Suitable F2:3 families for BSA will be selected based on their inoculation assay results.
   • During the spring cropping season this year, another inoculation assay was conducted using the parental lines of ‘TSS No. 3’ and several potentially resistant lines to identify an appropriate cross combination. Although lines ‘A’ and ‘4-1-2’ exhibited the highest levels of resistance, their horticultural characteristics, particularly their excessively long fruits, differ considerably from those of cultivated varieties. Because their relatively distant genetic relationship may reduce crossing success, the susceptible elite cultivar ‘TSS No. 3’ is planned to be crossed with the resistant line ‘111’.
2. Development of Molecular Markers to Assist Rice Seed Testing
   • Polymorphic molecular marker screening has been completed for the first batch of 10 rice varieties included in the study. These varieties are currently listed as recommended rice varieties by the Agriculture and Food Agency (AFA) (see Appendix for details). Among the 37 molecular markers tested, 25 exhibited polymorphic banding patterns and therefore have potential for use in rice varietal identification.
   • However, some of the tested varieties could not yet be effectively and accurately differentiated or clustered using these markers. Further studies will investigate the possible causes, and additional molecular markers will be tested and evaluated for their effectiveness in rice varietal identification.