Plant Cell Biotechnology Department, CSIR-Central Food Technological Research Institute, Mysore 570020, Karnataka, India.
Indian Institute of Science Education and Research, Bhopal, Bhauri, Madhya Pradesh 462066, India.
Int J Biol Macromol. 2022 Nov 1;220:942-953. doi: 10.1016/j.ijbiomac.2022.08.126. Epub 2022 Aug 20.
Serotonin N-acetyltransferase (SNAT) is the penultimate enzyme involved in plant melatonin biosynthesis. Identifying its expression under development and stress will reveal the regulatory role in the soybean. To identify and characterize SNAT, we employed genome-wide analysis, gene structure, cis-acting elements, expression, and enzyme activity. We identified seven putative genes by genome-wide analysis and found chloroplast signal peptides in three GmSNATs. To elucidate GmSNATs role, expression datasets of more than a hundred samples related to circadian rhythm, developmental stages, and stress conditions were analysed. Notably, the expression of GmSNAT1 did not show significant expression during biotic and abiotic stress. The GmSNAT1 sequence showed 67.8 and 72.2 % similarities with OsSNAT and AtSNAT, respectively. The Km and Vmax of the purified recombinant GmSNAT1 were 657 μM and 3780 pmol/min/mg, respectively. To further understand the GmSNAT1 role, we supplemented different concentrations of serotonin and melatonin to in-vitro cultures and seed priming. These studies revealed that the GmSNAT1 expression was significantly up-regulated at higher concentrations of serotonin and down-regulated at higher melatonin concentrations. We speculate that a high concentration of melatonin during abiotic, biotic stress, and in-vitro cultures are responsible for regulating GmSNAT1 expression, which may regulate them at the enzyme level during stress in soybean.
色氨酸 N-乙酰转移酶(SNAT)是植物褪黑素生物合成中倒数第二个酶。鉴定其在发育和胁迫下的表达情况,将揭示其在大豆中的调控作用。为了鉴定和表征 SNAT,我们采用了全基因组分析、基因结构、顺式作用元件、表达和酶活性分析。我们通过全基因组分析鉴定了七个假定的基因,并在三个 GmSNAT 中发现了质体信号肽。为了阐明 GmSNAT 的作用,我们分析了 100 多个与昼夜节律、发育阶段和胁迫条件相关的样本的表达数据集。值得注意的是,GmSNAT1 在生物和非生物胁迫下的表达没有显著变化。GmSNAT1 序列与 OsSNAT 和 AtSNAT 的相似性分别为 67.8%和 72.2%。纯化的重组 GmSNAT1 的 Km 和 Vmax 分别为 657 μM 和 3780 pmol/min/mg。为了进一步了解 GmSNAT1 的作用,我们在体外培养和种子引发中补充了不同浓度的血清素和褪黑素。这些研究表明,在较高浓度的血清素和较低浓度的褪黑素下,GmSNAT1 的表达显著上调。我们推测,在非生物、生物胁迫和体外培养期间,较高浓度的褪黑素可能负责调节 GmSNAT1 的表达,这可能在大豆胁迫期间在酶水平上调节它们。