Abstract
This study reported the isolation and in silico characterization of a full-length Xyloglucan endotransglycosylase/Hydrolase (XTH) cDNA from Neolamarckia cadamba, an important tropical light hardwood plantation tree species. XTH is considered as a key agent to regulate cell wall expansion and is believed to be responsible for the incorporation of newly synthesised xyloglucan into the wall matrix. The full-length of NcXTH was firstly predicted using the XTH singletons from the NcdbEST through contig mapping approach. Further validation and confirmation were conducted by amplifying the full-length XTH cDNA using RT-PCR approach. Two fulllength XTH cDNAs, namely NcXTH1 (JX134619) and NcXTH2 (JX134620) were discovered and the nucleotide sequences were 893 and 1,024 bp in length, respectively. The open reading frames for NcXTH1 and NcXTH2 were 858 and 915 bp, respectively. Results predicted that NcXTH1 and NcXTH2 proteins carry out XET activity but they are from different XTH family members. This full-length NcXTH cDNA can serve as good candidate genes in association genetics study which leads to Gene-Assisted Selection (GAS) in the N. cadamba tree breeding programme.
Highlights
Wood formation or known as xylogenesis is an ordered and complex developmental process in plants
Singleton Ncdx106G11 was longer than Ncdx016F05 and the alignment of these two sequences scored at 100 and three singletons, Ncdx106G11, Ncdx053B07 and Ncdx104G02, were selected for contig mapping to produce a hypothetical XTH1 cDNA sequence
The conserved genetic structural features were identifed in both Xyloglucan endotransglycosylase/Hydrolase (XTH) genes and confirmed the enzymatic role of NcXTH1 and NcXTH2
Summary
Wood formation or known as xylogenesis is an ordered and complex developmental process in plants. It involves cell division, cell expansion, secondary wall deposition, lignification and programmed cell death. Most of the enzymes that involved in cell wall biopolymers synthesis are under the Carbohydrate Active enzymes (CAZymes) family. These include Glycosyltransferases (GTs), Glycoside Hydrolases (GHs), Polysaccharide Lyases (PLs) and various Carbohydrate Esterases (CEs) (Geisler-Lee et al, 2006). XTH is considered as a key agent to regulate cell wall expansion and is believed to be responsible for the incorporation of newly synthesised xyloglucan into the wall matrix. In order for cell wall to expend, crosslinked or connections of microfibrils need to be broken
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