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The fruit of  Physalis  has a berry and a novelty called inflated calyx syndrome (ICS, also named the ‘Chinese lantern’). Elucidation of the underlying developmental mechanisms of fruit diversity demands an efficient gene functional inference platform. Here, we tested the application of the tobacco rattle virus (TRV)-mediated gene-silencing system in  Physalis floridana . First, we characterized the putative gene of a phytoene desaturase in  P. floridana  ( PfPDS ). Infecting the leaves of the  Physalis  seedlings with the  PfPDS - TRV  vector resulted in a bleached plant, including the developing leaves, floral organs, ICS, berry, and seed. These results indicated that a local VIGS treatment can efficiently induce a systemic mutated phenotype. qRT-PCR analyses revealed that the bleaching extent correlated to the mRNA reduction of the endogenous  PfPDS . Detailed comparisons of multiple infiltration and growth protocols allowed us to determine the optimal methodologies for VIGS manipulation in  Physalis . We subsequently utilized this optimized VIGS methodology to downregulate the expression of two MADS-box genes,  MPF2  and  MPF3 , and compared the resulting effects with gene-downregulation mediated by RNA interference (RNAi) methods. The VIGS-mediated gene knockdown plants were found to resemble the mutated phenotypes of floral calyx, fruiting calyx and pollen maturation of the RNAi transgenic plants for both  MPF2  and  MPF3 . Moreover, the two MADS-box genes were appeared to have a novel role in the pedicel development in  P. floridana . The major advantage of VIGS-based gene knockdown lies in practical aspects of saving time and easy manipulation as compared to the RNAi. Despite the lack of heritability and mosaic mutation phenotypes observed in some organs, the TRV-mediated gene silencing system provides an alternative efficient way to infer gene function in various developmental processes in  Physalis , thus facilitating understanding of the genetic basis of the evolution and development of the morphological diversities within the Solanaceae.

Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis