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      Study on the Profile of Capsanthin-Capsurobin Synthase (Ccs) Gene responsible for Carotenoid Synthesis in Chili Pepper (Capsicum frutescens L.) Mutants G1M6 M2 Generation

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      IOP Conference Series: Earth and Environmental Science
      IOP Publishing

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          Abstract

          The Capsanthin-capsorubin synthase (Ccs) is one of the enzymes that play a role in carotenoid pigment bio-synthesis. Mutations in the Ccs gene cause the formation of yellow or orange fruit colour instead of red. This study aims to analyze the genetic profile of the Ccs gene and its effect on the carotenoid content of the chili pepper mutant G1M6 M2generation. The profile of the Ccs gene was analyzed using Polymerase Chain Reaction (PCR) continued with sequencing methods.The carotenoid content was measured from chili fruit harvested at 35 and 65 days after planting. Comparison of the partial sequence of Ccs gene of the mutants and the Capsicum annuum var. Landrace, showed a substitution of adenine into guanine in the 124 bp nucleotide base sequence and the cytosine nucleotide substitution to thymine in the 556 bp nucleotide base sequence. However there was no difference between the Ccs gene sequence of the chili pepper G1M6/13, G1M6/7, G1M6/8 compared to the control plant (G1K1). The consistency showed by these sequence did not reflect the carotenoid content. Carotenoid content in the three mutant plants varied compared to control plants. The G1M6/8 mutant had higher carotenoid content, while the G1M6/13 mutant had a lower carotenoid content compared to the control plant. Since there was no differences on the partial sequence of the Ccs gene observed, the deviation of carotenoid content between mutant and control plants maybe governed by other part of the gene.

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          Clustal W and Clustal X version 2.0.

          The Clustal W and Clustal X multiple sequence alignment programs have been completely rewritten in C++. This will facilitate the further development of the alignment algorithms in the future and has allowed proper porting of the programs to the latest versions of Linux, Macintosh and Windows operating systems. The programs can be run on-line from the EBI web server: http://www.ebi.ac.uk/tools/clustalw2. The source code and executables for Windows, Linux and Macintosh computers are available from the EBI ftp site ftp://ftp.ebi.ac.uk/pub/software/clustalw2/
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            BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT

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              A comparison of the carotenoid accumulation in Capsicum varieties that show different ripening colours: deletion of the capsanthin-capsorubin synthase gene is not a prerequisite for the formation of a yellow pepper.

              Ripe pepper (Capsicum sp.) fruits can display a range of colours from white to deep red. To understand better the regulatory mechanisms of the carotenoid biosynthetic pathways that underlie these ripening colours, Capsicum varieties that show seven different fully ripe colour types were analysed. The levels and composition of the carotenoid accumulation in these samples at different stages of ripening were measured, and the resulting data were analysed in conjunction with the expression patterns of the carotenoid biosynthetic genes. It was found that red peppers accumulate increasing levels of total carotenoids during ripening, whereas non-red peppers accumulate lower levels of total carotenoids of varying composition. The expression levels of the phytoene synthase, phytoene desaturase, and capsanthin-capsorubin synthase (Ccs) genes are high in peppers with high levels of total carotenoid, whereas one or two of these genes are not expressed in peppers with lower levels of total carotenoid. Surprisingly, it was found that the Ccs gene is present in two Capsicum varieties whose ripe colour is yellow. This gene has never previously been shown to be present in yellow peppers. Sequence analyses of the Ccs gene further revealed two structural mutations in yellow peppers that may result in either a premature stop-codon or a frame-shift. Taken together with the fact that the Ccs transcript is not detectable in yellow peppers, our current results suggest that nonsense-mediated transcriptional gene silencing of Ccs and not the deletion of this gene is responsible for yellow ripening in Capsicum.
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                Author and article information

                Journal
                IOP Conference Series: Earth and Environmental Science
                IOP Conf. Ser.: Earth Environ. Sci.
                IOP Publishing
                1755-1307
                1755-1315
                December 01 2019
                December 01 2019
                : 391
                : 1
                : 012019
                Article
                10.1088/1755-1315/391/1/012019
                18920db8-05dc-41ca-88e5-556e137d955c
                © 2019

                http://creativecommons.org/licenses/by/3.0/

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