Silencing of the gibberellin receptor homolog, CsGID1a, affects locule formation in cucumber (Cucumis sativus) fruit

Abstract

SummaryGibberellins are phytohormones with many roles, including the regulation of fruit development. However, little is known about the relationship between GA perception and fleshy fruit ontogeny, and particularly locule formation.We characterized the expression of cucumber (Cucumis sativus) GA receptor gene (CsGID1a) using quantitative real-time PCR, in situ hybridization and a promoter::bglucuronidase (GUS) assay. CsGID1a-RNAi cucumber fruits were observed by dissecting microscope, scanning electron microscopy and transmission electron microscopy. Finally, genome-wide gene expression in young fruits from a control and the RNAi line was compared using a digital gene expression (DGE) analysis approach.The expression pattern of CsGID1a was found to be closely correlated with fruit locule formation, and silencing CsGID1a in cucumber resulted in fruits with abnormal carpels and locules. Overexpression of CsGID1a in the Arabidopsis thaliana double mutant (gid1a gid1c) resulted in 'cucumber locule-like' fruits. The DGE analysis suggested that expression of genes related to auxin synthesis and transport, as well as the cell cycle, was altered in CsGID1a-RNAi fruits, a result that was supported by comparing the auxin content and cellular structures of the control and transgenic fruits.This study demonstrates a previously uncharacterized GA signaling pathway that is essential for cucumber fruit locule formation.

Gibberellins modulate auxin responses during tomato (Solanum lycopersicum L.) fruit development

“…According to Lemaire‐Chamley et al (), the formation of the locular tissue in tomato requires the transduction of auxin signal that coordinates the enlargement of locular cells. Likewise, in cucumber fruits, CsGID1a (a GA receptor) would act as regulator of auxin synthesis and transport during the development of placental tissue (Liu et al ).…”

Cucumber (Cucumis sativus L.) Nitric Oxide Synthase Associated Gene1 (CsNOA1) Plays a Role in Chilling Stress

“…Samples were post-fixed with 1% osmic acid, washed in 0.1 M phosphate buffer (pH 6.8), dehydrated through an acetone series (using the sequence 30, 50, 70, 80, 90, and 100%), and then embedded in Spurr's resin. Thin cross-sections were made with a UC6I microtome (Leica) and examined with a JEM-123O scanning transmission electron microscope (Liu et al, 2016). …”

A Protocol for Agrobacterium-mediated Transformation of Cucumber (Cucumis sativus L.) from cotyledon explants

“…The average plant regeneration rate and transformation frequencies are affected by cucumber genotypes, explants types, explants preparation methods, the Agrobacterium infection methods, the concentration and ratio of ABA and BAP and culture methods ( Table 1, Table 2, Table 3, Table 4 and has been used to transform different cucumber varieties (Deltasta, Guonong25, cucumber line 3407, 3413, 3461, Chinses long: 461, 9930, R1407, R470, short 32x, and Xintaimici,) and obtained the transgenic plants successfully [25][26][27][28][29][30][31][32][33] . The pure line should be used when considering for obtain T 1 generation and for gene function analysis.…”

“…This protocol has improved the frequency of transformation to 26%, and we have successfully obtained over six hundred cucumber transgenic lines and published some papers of gene function analysis using this protocol [25][26][27][28][29][30][31][32][33] . We believe that this protocol is and will play important role in gene discovery, gene delivery, gene functional analyses and variety improvement for cucumber and…”

“…Our laboratory has established a rapid and efficient Agrobacterium-mediated transformation system for cucumber that differs from previously reported protocols, and we have identified critical factors that influence transformation efficiency. To date, we have produced more than 600 transgenic lines and functionally analyzed a range of cucumber genes using this protocol [25][26][27][28][29][30][31][32][33] . The method can be widely used in cucumber breeding, variety improvement, and the characterization of gene function, and can be a reference for the transformation of other cucurbits, such as Cucumis melo and C.metuliferus 21,23 .…”