Molecular cloning and expression analysis of the ethylene insensitive3 (EIN3) gene in cucumber (Cucumis sativus)

Abstract

ABSTRACT. The plant gaseous hormone ethylene regulates many aspects of plant growth, development, and responses to the environment. Ethylene insensitive3 (EIN3) is a key transcription factor involved in the ethylene signal transduction pathway. To gain a better understanding of this particular pathway in cucumber, the fulllength cDNA encoding EIN3 (designated as CsEIN3) was cloned from cucumber for the first time by rapid amplification of cDNA ends. The full length of CsEIN3 was 2560 bp, with an open reading frame of 1908 bp encoding 635 amino acids. Sequence alignment and phylogenetic analyses revealed that CsEIN3 has high homology with other plant EIN3/EIL proteins that were derived from a common ancestor during evolution, and CsEIN3 was grouped into a cluster along with melon. Homology modeling demonstrated that CsEIN3 has a highly similar structure to the specific DNA-binding domain contained in EIN3/EIL proteins. Based on quantitative reverse transcription-polymerase chain reaction analysis, we found that CsEIN3 was constitutively expressed in all organs examined, and was increased during flower development and maturation in both male and female flowers. Our results suggest that CsEIN3 is involved in processes of flower development. In conclusion, this study will provide the basis for further study on the role of EIN3 in relevant biological processes of cucumber and on the molecular mechanism of the cucumber ethylene signaling pathway.

Ectopic Expression of CsCTR1, a Cucumber CTR-Like Gene, Attenuates Constitutive Ethylene Signaling in an Arabidopsis ctr1-1 Mutant and Expression Pattern Analysis of CsCTR1 in Cucumber (Cucumis sativus)

“…The qRT-PCR was performed according to Bie et al (2013) [ 33 ]. A dissociation curve was performed to ensure the validity of each specific PCR product.…”

“…Compared with the ethylene biosynthesis pathway, very little is known about the role of ethylene signaling in the sex determination of flowers, mainly because the key components for the ethylene signaling pathway have not yet been identified in cucumber. So far, only a few of these components have been cloned from cucumber, including ETR2, ETR1, ERS and EIN3 [ 33 , 34 ]. Therefore, the majority of the key components remain to be isolated and characterized.…”

Investigation of the EIL/EIN3 Transcription Factor Gene Family Members and Their Expression Levels in the Early Stage of Cotton Fiber Development

“…The EIN3/EIL protein family is a small group of transcription factors in higher plants. Several transcription factor genes of EIL/EIN3 proteins have been investigated in Arabidopsis [16,17], tobacco [18], tomato [19], cucumber [20][21][22], and rice [23]. Plant specific EIL/EIN3 proteins have highly conserved amino acid sequences at the N-termini, and are located in the nuclei compartment [16].…”

A GAMYB homologue CsGAMYB1 regulates sex expression of cucumber via an ethylene-independent pathway

“…In this process, ethylene treatment can produce increased numbers of female flowers in cucumber (Iwahori et al , 1969; MacMurray and Miller, 1968), and the mechanism has been widely understood. Two major genes encoding ACC synthase (a key enzyme of ethylene biosynthesis), F ( CsACS1G ), and M ( CsACS2 ) control female sex expression in cucumber; the F gene governs the development of female flowers (Knopf and Trebitsh, 2006; Mibus and Tatlioglu, 2004; Trebitsh et al , 1997), whereas the M gene inhibits stamen development in flower buds (Bie et al , 2013; Li et al , 2012; Saito et al , 2007; Wang et al , 2010; Yamasaki et al , 2001; Yamazaki et al , 2003). In addition, GA application can promote the male tendency (Pike and Peterson, 1969), but the molecular regulation remains elusive.…”