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The strawberry ( Fragaria  ×  ananassa ) is one of the most preferred fresh fruit worldwide, accumulates numerous flavonoids but has limited shelf life due to excessive tissue softening caused by cell wall degradation. Since lignin is one of the polymers that strengthen plant cell walls and might contribute to some extent to fruit firmness monolignol biosynthesis was studied in strawberry fruit. Cinnamoyl-CoA reductase ( CCR ), cinnamyl alcohol dehydrogenase ( CAD ), and a peroxidase ( POD27 ) gene were strongly expressed in red, ripe fruit whereas a second  POD  gene was primarily expressed in green, immature fruit. Moreover,  FaPOD 27 transcripts were strongly and constitutively induced in fruits exposed to  Agrobacterium  infection. Gene expression levels and enzymatic activities of FaCCR and FaCAD were efficiently suppressed through RNAi in  FaCCR- and FaCAD -silenced strawberries. Besides, significantly elevated  FaPOD  transcript levels were detected after agroinfiltration of pBI- FaPOD  constructs in fruits. At the same time, levels of G-monomers were considerably reduced in  FaCCR -silenced fruits whereas the proportion of both G- and S-monomers decisively decreased in  FaCAD -silenced and pBI- FaPOD  fruits. Development, firmness, and lignin level of the treated fruits were similar to pBI-intron control fruits, presumably attributed to increased expression levels of  FaPOD27  upon agroinfiltration. Additionally, enhanced firmness, accompanied with elevated lignin levels, was revealed in chalcone synthase-deficient fruits (CHS − ), independent of down- or up-regulation of individual and combined  FaCCR .  FaCAD , and  FaPOD  genes by agroinfiltration, when compared to CHS − /pBI-intron control fruits. These approaches provide further insight into the genetic control of flavonoid and lignin synthesis in strawberries. The results suggest that  FaPOD27  is a key gene for lignin biosynthesis in strawberry fruit and thus to improving the firmness of strawberries.

FaPOD27 functions in the metabolism of polyphenols in strawberry fruit (Fragaria sp.)