Abstract

Gigantochloa levis is an economically important tropical bamboo; however, molecular studies on this species remain limited due to difficulties in obtaining high-quality RNA and the scarcity of genomic resources. This study aimed to develop an optimized RNA extraction protocol and generate a de novo transcriptome assembly for G. levis. Leaf, shoot, and stem tissues were processed using a modified TransZol Up Plus RNA Kit, and RNA quality was evaluated by spectrophotometry, agarose gel electrophoresis, and Agilent bioanalyzer analysis. High-quality RNA was subsequently sequenced using the Illumina NovaSeq 6000 platform. Sequencing generated 57.6 GB of data with more than 98% clean reads, low error rates, and high base-calling accuracy. De novo assembly using Trinity produced 1,056,042 contigs, providing a comprehensive transcriptomic dataset for the species. The optimized extraction protocol consistently yielded RNA integrity numbers above 8.0 across all tissues. Benchmarking universal single-copy orthologs (BUSCO) analysis demonstrated high transcriptome completeness, with 90.7% and 90.6% complete BUSCOs identified in the Trinity and CD-HIT-EST transcript sets, respectively. The CD-HIT-EST assembly also showed reduced duplicated BUSCOs and increased single-copy BUSCOs, indicating effective removal of redundant transcripts. All sequencing data have been deposited in the National Center for Biotechnology Information Sequence Read Archive. Collectively, this study establishes a reliable RNA extraction workflow and provides a valuable transcriptomic resource that will facilitate future functional genomics, comparative transcriptomics, and plant biotechnology research in bamboo and other lignocellulosic species.

Key words: Bamboo transcriptomics, de novo transcriptome assembly, Gigantochloa levis, RNA extraction, RNA sequencing