Deciphering Molecular Pathways of Bletilla striata Seeds Symbiotic Germination with Tulasnella sp. bj1

Orchid seed germination requires symbiotic association with mycorrhizal fungi that provide essential nutrients for germination and subsequent growth. Extensive research has elucidated the pivotal role of the mycorrhizal fungus <i>Tulasnella</i> sp. in the modulation of seed germination and growth processes in <i>Bletilla striata</i> (Thunb.) Reiehb.f. However, the molecular mechanisms underlying this symbiosis remain poorly characterized. Our integrated transcriptomic-metabolomic analysis of symbiotic germination revealed that co-cultivation of <i>Tulasnella</i> sp. bj1 with <i>B. striata</i> seeds significantly downregulates the expression of plant-derived flavonoid biosynthetic genes, with flavonoid degradation potentially alleviating germination and growth inhibition. The bj1 strain modulates indoleacetic acid (IAA) biosynthesis in <i>B. striata</i> by upregulating the expression of plant-derived tryptophan decarboxylase (TDC) in the tryptophan pathway and hydrolytic enzymes (NtAMI) in the indoleacetamide pathway, with elevated IAA potentially contributing to seed germination and growth. Moreover, bj1 suppresses the jasmonic acid (JA) biosynthetic pathway of <i>B. striata</i> by downregulating key plant-derived biosynthetic genes, concurrently promoting the accumulation of 12-hydroxyjasmonic acid-a metabolite associated with plant immune regulation that may favor colonization and symbiotic establishment with <i>B. striata</i> seeds. Additionally, bj1 induces the expression of polysaccharide-degrading enzymes, potentially improving carbon source utilization to support protocorm development. In conclusion, bj1 modulates the immune response of <i>B. striata</i> seeds, facilitating the establishment of a symbiotic relationship. Subsequently, the germination and growth of <i>B. striata</i> seeds are enhanced through reduced flavonoid accumulation, increased IAA synthesis, and improved carbon source utilization. Consequently, this investigation provides a crucial foundation for elucidating mechanisms governing symbiotic germination in <i>B. striata</i>.