Lin Chun, Huang Qiuqiu, Liu Zhengjie, Brown Sylvia E, Chen Qing, Li Yuping, Dong Yumei, Wu He, Mao Zichao
College of Agronomy and Biotechnology, Yunnan Agricultural University (YNAU), Kunming, Yunnan 650201, China; Institute of Improvement and Utilization of Characteristic Resource Plants (YNAU), Kunming, China; The Laboratory for Crop Production and Intelligent Agriculture of Yunnan Province, Kunming, China.
College of Agronomy and Biotechnology, Yunnan Agricultural University (YNAU), Kunming, Yunnan 650201, China.
Gene. 2023 Apr 30;862:147284. doi: 10.1016/j.gene.2023.147284. Epub 2023 Feb 11.
The full length CDS of an A20 and AN1 type zinc finger gene (named AoSAP8-P), located nearby the male specific Y chromosome (MSY) region of Asparagus officinalis (garden asparagus) was amplified by RT-PCR from purple passion. This gene, predicted as the stress associated protein (SAPs) gene families, encodes 172 amino acids with multiple cis elements including light, stress response box, MYB and ERF binding sites on its promoter. To analyze its function, the gene expression of different organs in different asparagus gender were analyzed and the overexpressed transgenic Nicotiana sylvestris lines were generated. The results showed the gene was highly expressed in both flower and root of male garden asparagus; the germination rate of seeds of the T2 transgenic lines (T2-5-4 and T2-7-1) under the stress conditions of 125 mM NaCl and 150 mM mannitol were significantly higher than the wild type (WT) respectively. When the potted T2-5-4, T2-7-1 lines and WT were subjected to drought stress for 24 days and the leaf discs immerged into 20 % PEG6000 and 300 mM NaCl solution for 48 h respectively, the T2-5-4 and T2-7-1 with AoSAP8-P expression showed stronger drought, salt and osmotic stress tolerance. When compared, the effects of AoSAP8-P overexpression on productive development showed that the flowering time of transgenic lines, were ∼ 9 day earlier with larger but fewer pollens than its WT counterparts. However, there were no significant differences in anthers, stigmas and pollen viability between the transgenic lines and WT. Our results suggested that, the AoSAP8-P gene plays a role in improving the stress resistance and shortening seeds generation time for perianal survival during the growth and development of garden asparagus.
从紫芦笋中通过RT-PCR扩增出芦笋（石刁柏）雄性特异性Y染色体（MSY）区域附近的一个A20和AN1型锌指基因（命名为AoSAP8-P）的全长CDS。该基因被预测属于胁迫相关蛋白（SAPs）基因家族，编码172个氨基酸，其启动子上有多个顺式元件，包括光、胁迫响应框、MYB和ERF结合位点。为了分析其功能，对不同性别芦笋不同器官的基因表达进行了分析，并构建了过表达的转基因野生烟草株系。结果表明，该基因在雄性芦笋的花和根中均高表达；在125 mM NaCl和150 mM甘露醇胁迫条件下，T2转基因株系（T2-5-4和T2-7-1）种子的发芽率分别显著高于野生型（WT）。当盆栽的T2-5-4、T2-7-1株系和WT分别遭受24天干旱胁迫，以及叶片分别浸入20% PEG6000和300 mM NaCl溶液中48小时时，表达AoSAP8-P的T2-5-4和T2-7-1表现出更强的干旱、盐和渗透胁迫耐受性。相比之下，AoSAP8-P过表达对生产发育的影响表明，转基因株系的开花时间比野生型早约9天，花粉更大但数量更少。然而，转基因株系和野生型之间的花药、柱头和花粉活力没有显著差异。我们的结果表明，AoSAP8-P基因在芦笋生长发育过程中，对提高抗逆性和缩短种子世代时间以利于周年存活发挥作用。
