Rhamnolipid-Enriched PA3 Fraction from Pseudomonas aeruginosa SWUC02 Primes Chili Plant Defense Against Anthracnose

Chili anthracnose, caused by <i>Colletotrichum truncatum</i>, causes significant yield loss in chili production. In this study, we investigated the elicitor properties of a rhamnolipid (RL)-enriched PA3 fraction derived from <i>Pseudomonas aeruginosa</i> SWUC02 in inducing systemic resistance in yellow chili seedlings and antifungal activity against <i>C. truncatum</i> CFPL01 (Col). Fractionation of the ethyl acetate extract yielded 12 fractions, with PA3 demonstrating the most effective disease suppression, reducing the disease severity index to 4 ± 7.35% at 7 days post-inoculation compared with Col inoculation alone (83 ± 23.57%). PA3 also exhibited direct antifungal activity, inhibiting Col mycelial growth by 41 ± 0.96% at 200 µg/mL. Subfractionation revealed PA3 as a mixture of mono- and di-RLs, confirmed by ¹H nuclear magnetic resonance and electrospray ionization mass spectrometry data. Additionally, PA3 enhanced seed germination and promoted plant growth without causing phytotoxicity. Transcriptomics revealed that PA3 pre-treatment prior to Col infection primed the defense response, upregulating defense-related genes involved in the phenylpropanoid, flavonoid, and jasmonic acid biosynthesis pathways, as well as those associated with cell wall reinforcement. Our findings highlight the potential of RL-enriched PA3 as both an antifungal agent and a plant defense elicitor, with transcriptome data providing new insights into defense priming and resistance pathways in chili, offering an eco-friendly solution for sustainable anthracnose management.