Abstract
Promoters are specified segments of DNA that lead to the initiation of transcription of a specific gene. The designing of a gene cassette for plant transformation is significantly dependent upon the specificity of a promoter. Constitutive Cauliflower mosaic virus promoter, CaMV35S, due to its developmental role, is the most commonly used promoter in plant transformation. While Gossypium hirsutum (Gh) being fiber-specific promoter (GhSCFP) specifically activates transcription in seed coat and fiber associated genes. The Expansin genes are renowned for their versatile roles in plant growth. The overexpression of Expansin genes has been reported to enhance fiber length and fineness. Thus, in this study, a local Cotton variety was transformed with Expansin (CpEXPA1) gene, in the form of two separate cassettes, each with a different promoter, named as 35SEXPA1 and FSEXPA1 expressed under CaMV35S and GhSCFP promoters respectively. Integration and Spatiotemporal relative expression of the transgene were studied in an advanced generation. GhSCFP bearing transgene expression was significantly higher in Cotton fiber than other plant parts. While transgene with CaMV35S promoter was found to be continually expressing in all tissues but the expression was lower in fiber than that expressed under GhSCFP. The temporal expression profile was quite interesting with a gradual increasing pattern of both constructs from 1DPA (days post anthesis) to 18DPA and decreased expression from 24 to 30 DPA. Besides the relative expression of promoters, fiber cellulose quantification and fluorescence intensity were also observed. The study significantly compared the two most commonly used promoters and it is deduced from the results that the GhSCFP promoter could be used more efficiently in fiber when compared with CaMV35S which being constitutive in nature preferred for expression in all parts of the plant.
Highlights
The cotton is the backbone of the global textile industry
All the work is conducted at Center of Excellence in Molecular Biology (CEMB), University of the Punjab under controlled conditions
The in-silico joining of NCBI retrieved Expansin-like A1 gene sequence with two different promoters was done i.e. constitutive Cauliflower Mosaic Virus, CamV35S, [14] and Gossypium hirsutum seed coat Fiber-specific protease promoter, GhSCFSP, [15] with final gene constructs as 35SEXPA1 and FSEXPA1 respectively (Fig 1A and 1B)
Summary
The cotton is the backbone of the global textile industry. More than fifty-five countries are dealing in cotton cultivation, fiber processing, and export. Pakistan is the fourth-largest consumer and exporter of Cotton [1]. It comes in the fifth position among the largest Cotton producing countries of world and third position for having the largest capacity of raw fiber spinning in the world [2]. Several efforts have been made to improve the fiber quality and yield to strengthen.
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