Sains Malaysiana 45(5)(2016): 717–727

Isolation and Characterization of Full-Length Cellulose Synthase Gene (HsCesA1) from Roselle (Hibiscus sabdariffa L. var. UMKL)

(Pengasingan dan Pencirian Panjang Penuh Gen Selulosa Sintase (HsCesA1) daripada Rosel (Hibiscus sabdariffa L. var. UMKL))

SEYEDEH SAREH SEYEDI¹, SOON GUAN TAN¹, PARAMESWARI NAMASIVAYAM¹

 & CHRISTINA SEOK YIEN YONG²*

¹Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences

Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia

²Department of Biology, Faculty of Science, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia

Diserahkan: 18 Mac 2015/Diterima: 23 November 2015

ABSTRACT

The Hibiscus sabdariffa var. UMKL (Roselle) investigated here may potentially be used as an alternative fibre source. To the best of our knowledge, there was no study focusing on the genetics underlying the cellulose biosynthesis machinery in Roselle thus far. This paper presents the results of the first isolation of the cellulose synthase gene, HsCesA1 from this plant, which is fundamental for working towards understanding the functions of CesA genes in the cellulose biosynthesis of Roselle. A full-length HsCesA1 cDNA of 3528 bp in length (accession no: KJ608192) encoding a polypeptide of 974 amino acid was isolated. The full-length HsCesA1 gene of 5489 bp length (accession no: KJ661223) with 11-introns and a promoter region of 737 bp was further isolated. Important and conserved characteristics of a CesA protein were identified in the HsCesA1 deduced amino acid sequence, which strengthened the prediction that the isolated gene being a cellulose synthase belonging to the processive class of the 2-glycosyltransferase family 2A. Relative gene expression analysis by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) on young leaf and stem tissues found that HsCesA1 had similar levels of gene expression in both tissues. Phylogenetic and Blast analyses also supported the prediction that the isolated HsCesA1 may play roles in the cell wall depositions in both leaf and stem tissues.

Keywords: Cellulose; cellulose synthase gene (CesA); fibre; HsCesA1; promoter; semi-quantitative RT-PCR

ABSTRAK

Hibiscus sabdariffa var. UMKL (Rosel) yang dikaji berpotensi digunakan sebagai sumber serat alternatif. Sepanjang pengetahuan kami, tidak ada kajian memberi tumpuan kepada genetik asas jentera selulosa biosintesis dalam Roselle setakat ini. Kertas ini membentangkan keputusan pengasingan pertama gen selulosa sintase, HsCesA1 daripada tumbuhan ini, yang merupakan asas ke arah memahami fungsi gen CesA dalam biosintesis selulosa Roselle. Satu jujukan penuh cDNA HsCesA1 dengan panjang 3528 bp (No aksesi: KJ608192) mengekodkan polipeptida sepanjang 974 asid amino telah dipencilkan. Jujukan penuh gen HsCesA1 sepanjang 5489 bp (No. aksesi: KJ661223) dengan 11-intron dan rantau promoter sepanjang 737 bp juga telah dipencilkan. Ciri penting dan abadi protin CesA telah dikenal pasti di dalam jujukan asid amino mendeduksi HsCesA1, yang mengukuhkan ramalan bahawa gen yang terpencil adalah selulosa sintase yang tergolong dalam kelas prosesif 2-glycosyltransferase keluarga 2A. Analisis relatif ekspresi gen menggunakan semi-kuantitatif tindak balas polimeras berantai transkripsi berbalik (RT-PCR) pada daun muda dan batang tisu mendapati HsCesA1 mempunyai tahap ekspresi gen yang sama dalam kedua-dua tisu. Analisis filogeni dan Blast juga menyokong ramalan bahawa pencilan HsCesA1 mungkin memainkan peranan dalam endapan dinding sel dalam kedua-dua tisu daun dan batang.

Kata kunci: Gen selulosa sintase (CesA); HsCesA1; promoter; selulosa; semi-kuantitatif RT-PCR; serat

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