Cloning And Expression Of F-Box Protein Genes, Carf-Box1, Carf-Box2in Chickpea(Cicer Arietinum L.)

F-box proteins play important roles in metabolism, cell-cycle control, transcription control, cell differentiation and apoptosis, signal transduction via the ubiquitin pathway. Many results suggest that F-box protein in SCF complexes regulate many phytohormones signalling pathways, including the auxin, jasmonate, gibberellin and ethylene pathways. Moreover, F-box proteins are also involved in responses to abiotic stresses such as drought, cold and high salinity. The study on F-box proteins is important to the elucidation the mechanism of the plant responses to the stress. Chickpea (C. arietinum L.) is the third important legume crop grown mainly in the arid and semi-arid regions. It is an annual plant with a short life, a small genome size. Through long-term evolution and adaptation to extreme conditions, chickpea has been found to be rich in resistance genes for a range of abiotic stresses, including drought, cold and high salinity, and has been suggested as a model plant for investigation of physiological mechanisms of plant development and responses to stresses. However, chickpea is scarcely studied in the molecular biology.Herein, two F-box genes were picked out from the cDNA library, and cloned by RACE technology. Bioinformatic and expression profile analyses were conducted to further understand the molecules and their function in many crucial biological processes including growth and development, stress response, signal transduction and disease resistance.The F-box protein gene CarF-box1was cloned by rapid amplification of cDNA ends (RACE). The full cDNA sequence of CarF-box1is1093bp and encoded a38.37kDa protein with345amino acids. The protein contains the F-box domain and Kelch domain. Its DNA sequence has no intron. Real-time quantitative PCR analysis showed that CarF-box1was expressed in root, stem, leaf, flower, mature seed and mature pod of chickpea, and especially high in flower. The expression of CarF-box1is increased gradually during germination. Semi-quantitative RT-PCR and real-time q quantitative PCR analysis indicated that the expression of CarF-box1could be induced by drought in leaf and salt in root significantly. And under the treatment of IAA, the expression of CarF-box1has increasing too. Meanwhile, transient expression of the CarF-box1::GFP protein in onion epidermal cells showed that CarF-box1protein was localized in cell nuclei. Our results suggest that CarF-box1may play multiple roles in plant growth and abiotic resistance pathways via the ubiquitin pathway. As well as CarF-box1has some effect on the development of flowers and the flowing time.The full cDNA sequence of CarF-box2is1201bp and encoded a29.13kDa protein with261amino acids. The protein contains the F-box domain and LysM domain. Its DNA sequence has a684bp intron. Real-time quantitative PCR analysis showed that CarF-box1was expressed in root, stem, leaf, flower, mature seed and mature pod of chickpea, and especially low in mature pod. The expression of CarF-box2is increased generally during germination. Semi-quantitative RT-PCR and real-time quantitative PCR analysis indicated that the expression of CarF-box2could be induced by salt, cold, ABA and GA significantly in the leaf. CarF-box2could be significantly induced by drought, GA, IAA and ABA in the root. Meanwhile, transient expression of the CarF-box2::GFP protein in onion epidermal cells showed that CarF-box2protein was localized in cell nuclei. Our results suggest that CarF-box2may play multiple roles in plant growth and abiotic resistance pathways.