Expansins are cell-wall-loosening proteins that have multiple roles during plant development and stress- related processes. In this study, a novel expansin gene PwEXP2 was cloned by rapid amplification of cDNA ends based on the cDNA library of Picea wilsonii and EST fragment of PwEXP2. It was found that PwEXP2 coded 253 amino acids, and putative signal peptides exist at the N-terminal, followed by 8 cysteines, a HFD (His-Phe-Asp) conserved domain, and 4 tryptophan residues at the C-terminal. PwEXP2 was located in cytoplasm and nucleus when transformed in an onion epidermal cell. Quantitative real-time PCR assays showed that PwEXP2 was expressed in various tissues with a relatively high level in needles and low level in mature pollen. The expression level of PwEXP2 dramatically increased after seed germination. Gene expression profiles in abiotic stresses showed that PwEXP2 was induced by high temperature and osmotic stress but not involved in ABA-dependent signaling path- way. These results display the important roles of the PwEXP2 in plant development and multiple adversity stresses.
从青杄的cDNA文库中筛选出一个生长素抑制蛋白基因PwARP-1(Auxin-repressed protein 1),并通过RACE-PCR技术获得PwARP-1 cDNA全长。生物信息学分析显示,PwARP-1基因编码155个氨基酸残基的蛋白,分子量为17.1 kD,理论等电点为10.07,富含无规则卷曲。以多年生青杄和青杄幼苗为研究材料,通过RT-qPCR技术检测PwARP-1的组织特异性表达、激素的响应模式及种子萌发过程中的表达量模式。结果表明,PwARP-1在青杄各个组织中均有表达,在花粉中表达量最高,在种子中的表达量最低。PwARP-1在种子萌发过程中表达量呈先上升,在第10天表达量达到最高后开始下降。进一步试验表明,PwARP-1能响应多种逆境胁迫和激素处理。在赤霉素(GA)和生长素(IAA)处理下PwARP-1的表达量被抑制。在干旱胁迫和油菜素内酯(BR)激素处理下PwARP-1的表达量逐渐升高,而在在盐胁迫、茉莉酸甲酯(MeJA)和脱落酸(ABA)等激素处理下PwARP-1的表达量先下降后上升。这些结果显示PwARP-1可能参与了植物种子萌发、多种逆境胁迫和激素响应过程。