Cloning and Expression Analysis of Chitinase Gene in Hami Melon
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Abstract:
Chitinase, an important pathogenesis-related protein, plays an active role in disease resistance in plants. In order to explore the resistance of chitinase in Hami melon to Penicillium, real-time polymerase chain reaction (RT-PCR) was used to clone the cDNA of chitinase, and relative fluorescence quantitative PCR (SYBR Green) was used to analyze its expression. A gene fragment of 1153 bp was cloned, and was denoted as HmCHI. The largest open reading frame in HmCHI was 445 nucleotides long, encoded 145 amino acids, and belonged to Class III of Family 18 chitinase genes. Using software prediction, the encoded protein was determined to be a hydrophobic non-transmembrane protein distributed within the intercellular space, with a molecular weight of 15.04 ku, isoelectric point of 8.82, and one signal peptide. The amino acid sequence of HmCHI had the closest similarity with Cucumis sativus (XP_004145916.1). Fluorescence quantitative analysis showed that the expression peak in the experimental group appeared later and was maintained for a longer time, and that the magnitude of upregulation was significantly higher than the control. These results indicated that HmCHI in the Hami melon plays an important role in fungal stress response.
