## 2 Materials and Methods
### Retrieval of Protein Sequences and In Silico Analysis  
The amino acid sequences of LCYB and CGP from _Dunaliella sp_. were retrieved from the protein database of the National Center for Biotechnology Information (NCBI, [[http://www.ncbi.nlm.nih.gov/protein/](http://www.ncbi.nlm.nih.gov/protein/)]([http://www.ncbi.nlm.nih.gov/protein/](http://www.ncbi.nlm.nih.gov/protein/))) under the accession numbers of ACA34344.1 and AFU62958.1, respectively. The nucleotide sequence of SUMO (small ubiquitin-related modifier) was searched by blast against the Expressed Sequence Tag (EST) database of _Dunaliella_ sp. Then, the open reading frame (ORF) sequences were generated from the corresponding cDNA (GenBank: HO848823.1) using ORFfinder from NCBI, and the obtained proteins were blasted against the non-redundant protein database. The protein sequence corresponding to ORF2 presented 52.87% and 49.47% with SUMO protein from two Chlorophyta _Ostreococcus tauri_ and _Monoraphidium neglectum_, respectively. The motifs search was performed using the motif search tools from KEGG (Kyoto Encyclopedia of Genes and Genomes) ([[https://www.genome.jp/tools/motif/](https://www.genome.jp/tools/motif/)]([https://www.genome.jp/tools/motif/](https://www.genome.jp/tools/motif/))). \begin{table}
\begin{tabular}{c c c c c c c c} \hline \hline
**Gene Name** & **Abbreviation** & \multicolumn{3}{c}{**Primer Sequence (F\(>\)3\({}^{\prime}\))**} & **Amplion Size (bp)** & **Tm (\({}^{\circ}\)C)** & **E (\%)** & **Reference/Access Number** \\ \hline \multirow{2}{*}{_Actin_} & _Act_ & F & ACCCACACACCTTCTTCAACAGG & \multirow{2}{*}{150} & 86.76 & 111.35 & [39] \\  & & R & GCATGGCTACATACATGGCA & & & \\ \hline \multirow{2}{*}{_185 RNA_} & _185_ & F & TIGGTGATGGCGCGTGTGT & - & 83.2 & 92.6 & [40] \\  & & R & GCCCGCTCTCTCTCTCT & & & \\ \hline \multirow{2}{*}{_Alpha tubulin_} & \multirow{2}{*}{_a-TUB_} & F & GAGCATACACACACCCCCCTTGA & \multirow{2}{*}{105} & 86.03 & \multirow{2}{*}{106.2} & \multirow{2}{*}{JF346103.1} \\  & & R & CACCACCACCACCGGCTACAT & & & \\ \hline \multirow{2}{*}{_Beta tubulin_} & \multirow{2}{*}{_\beta-TUB_} & F & AGGAGGGCGCAGTTTGAGGGT & \multirow{2}{*}{137} & \multirow{2}{*}{83.12} & \multirow{2}{*}{92.4} & \multirow{2}{*}{DQ80916.1} \\  & & R & AGGCCGGTGGACATCTGAACAG & & & \\ \hline \multirow{2}{*}{_Enolase_} & _ENO_ & F & TGAAGGGCCTGAGCCCAG & \multirow{2}{*}{123} & \multirow{2}{*}{85.66} & \multirow{2}{*}{97.23} & \multirow{2}{*}{KM008612.1} \\  & & R & TCGCGATTGCGCAGGAGCA & & & \\ \hline \multirow{2}{*}{_1-deoxy-D-xylulose_} & \multirow{2}{*}{_DXS_} & F & AGCATATACGCCTGGCACAT & \multirow{2}{*}{102} & \multirow{2}{*}{85.67} & \multirow{2}{*}{94.86} & \multirow{2}{*}{FJ469276.1} \\  & & R & GCACACACACAGGCCAGCTT & & & \\ \hline \multirow{2}{*}{_Lycopene \(\beta\)-cyclase_} & \multirow{2}{*}{_LCYB_} & F & CCTGCACATACACACAGGCG & \multirow{2}{*}{130} & \multirow{2}{*}{87.91} & \multirow{2}{*}{97.1} & \multirow{2}{*}{EU327876.1} \\  & & R & GGGGGTTTTCGGGCTCTCT & & & \\ \hline \multirow{2}{*}{_Cardenet globule_} & \multirow{2}{*}{_CGP_} & F & GCGACACACAGCAGCAGCTACACAC & \multirow{2}{*}{104} & \multirow{2}{*}{85.18} & \multirow{2}{*}{111} & \multirow{2}{*}{JF3466677.1} \\  & & R & GAGACTCGCGCGACACAC & & & \\ \hline \multirow{2}{*}{_Chloroplast-localized_} & \multirow{2}{*}{_HSP70_} & F & TCGACGCTGGTGTGTGTGT & \multirow{2}{*}{124} & \multirow{2}{*}{87.88} & \multirow{2}{*}{99.03} & \multirow{2}{*}{AJ271605.2} \\  & & R & AGGGTGGTGTGTGGGGGTGTGT & & & \\ \hline \multirow{2}{*}{_Chloroplast tribulose_} & \multirow{2}{*}{_RPE_} & F & TGAACGACCTGGCGTCCA & \multirow{2}{*}{120} & \multirow{2}{*}{85.96} & \multirow{2}{*}{97.33} & \multirow{2}{*}{JF346108.1} \\  & & R & AGCCAGGTTCACGGACATG & & & \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Primers used in this study and their corresponding amplicons size, melting temperature Tm (\({}^{\circ}\)C) based on melt curve analysis, amplification efficiency E (%), and their reference or access number.