Loss-of-function ethylene insensitive 2 (EIN2) mutations showed ethylene insensitivity in Arabidopsis, which indicated an essential role of EIN2 in ethylene signaling. However, the function of EIN2 in fruit ripening has not been investigated. To gain a better understanding of EIN2, the temporal regulation of LeEIN2 expression during tomato fruit development was analyzed. The expression of LeEIN2 was constant at different stages of fruit development, and was not regulated by ethylene. Moreover, LeEIN2-silenced tomato fruits were developed using a virus-induced gene silencing fruit system to study the role of LeEIN2 in tomato fruit ripening. Silenced fruits had a delay in fruit development and ripening, related to greatly descended expression of ethylene-related and ripening-related genes in comparison with those of control fruits. These results suggested LeEIN2 positively mediated ethylene signals during tomato development. In addition, there were fewer seeds and locules in the silenced fruit than those in the control fruit, like the phenotype of parthenocarpic tomato fruit. The content of auxin and the expression of auxin-regulated gene were declined in silenced fruit, which indicated that EIN2 might be important for crosstalk between ethylene and auxin hormones. doi: 10.1111/j. 1672-9072.2006.00366.x; available online at www.blackwell-synergy.com, www.jipb.netThe plant hormone ethylene has a profound impact on plant growth and development, including fruit ripening (Abeles et al. 1992). Most components of the ethylene signal-transduction pathway have been identified in Arabidopsis thaliana (Kieber 1997;Guo and Ecker 2004). Ethylene is perceived by a family of receptors, and then the signal is transduced to the constitutive triple response (CTR), a protein kinase as the next component of the signal transduction pathway. Ethylene insensitive 3 (EIN3), the downstream of CTR, ultimately leads to the regulation of gene expression (Guo and Ecker 2004). EIN2, which acts as the downstream of CTR1 and the upstream of EIN3, plays an essential role in ethylene signal transduction (Chen and Bleecker 1995;Shibuya et al. 2004). In addition, EIN2 is also a cross-talk point between multiple hormone signaling pathways (Alonso et al. 1999;Adams-Phillips et al. 2004).Ethylene has a great impact on climacteric fruit ripening by coordinating the expression of genes (Alexander and Grierson 2002). The tomato is a typical climacteric fruit whose ripening is regulated by ethylene. Never ripe (Nr) tomato mutants that contain a mutation in the ethylene-binding domain of the NR receptor produce non-ripening fruits (Wilkinson et al. 1995). The delaying of fruit ripening is also observed in tomato plants with reduced expression of the tomato EIN3, like genes (LeEIL1, LeEIL2, and LeEIL3) (Fu et al. 2005). In addition, the overexpression of LeEIL1 can partially restore ripening in the Nr tomato mutant (Chen et al. 2004). This suggests that all Silencing of LeEIN2 Alters Fruit Ripening 1479 components of ethylene signaling might play an...