Cloning and Expression of the TgPUX10 Gene from Torreya grandis Pollen
Using cDNA synthesized from RNA extracted from Torreya grandis pollen as a template,a PUX10-like cDNA se-quence was isolated through RT-PCR,designated as TgPUX10.This gene was subsequently identified,cloned,and sub-jected to expression analysis.The results showed that the complete open reading frame(ORF)of the TgPUX10 gene is 1437 bp,encoding a protein consisting of 477 amino acid residues.The amino acid sequence of the TgPUX10 protein con-tains functional domains including UBA,HP,UAS,and UBX.By constructing a transient expression vector and utilizing an Arabidopsis suspension cell transformation system,it was found that TgPUX10 protein localizes to lipid body organelles.Furthermore,during the pollen germination stages of 0,2,and 4 days,the expression levels of lipid body-related genes TgOLE and TgCLO showed a declining trend,while the expression level of TgPUX10 significantly increased.This indicates that TgPUX10 influences pollen germination in T.grandis and is involved in the degradation process of lipid bodies.Tg-PUX10 exhibited relatively conserved expression across different varieties of T.grandis.Compared to other varieties,T.grandis'Xifei'exhibited higher pollen viability,with a germination rate reaching 35.5%.
万方数据
维普
