Subjects: Biology >> Botany >> Applied botany submitted time 2022-03-30 Cooperative journals: 《广西植物》
Abstract： Late embryogenesis abundant (LEA) is closely related to plant resilience, protects plant cells, reduces plant damage under drought stress, and is widely present in organisms. LEA proteins are less studied in ferns, and Selaginella pulvinata is a fern with the ability to survive drought stress, and it is a fern with a strong recovery ability under drought stress. To investigate the molecular mechanisms and expression characteristics of the SpLEA1 gene in drought-tolerant plants, this study used the highly drought-tolerant plant S. pulvinata as experimental material and obtained the cDNA sequence of the SpLEA1 gene by RT-PCR based on the transcriptome sequencing results. The promoter sequence was obtained by the HiTail-PCR technique, and the sequence was analyzed by bioinformatics. qRT-PCR was used to analyze the expression pattern of the SpLEA1 gene under drought stress. The results were as follows: (1) The length of SpLEA1 was 476 bp, the open reading frame (ORF) was 279 bp, and it encoded 92 amino acids. The predicted molecular weight of the protein was 9 491.46 Da, and the isoelectric point was 5.45. The predicted protein structure analysis showed that the protein was hydrophilic. The protein contained 10 phosphorylation sites, of which 6 serine, 3 tyrosine, and 1 threonine, respectively, and the predicted secondary and tertiary structures showed that the protein was mainly composed of α-helix and random coil. (2) The conserved structural domain of the SpLEA1 protein was predicted to be Lea-5, derived from the LEA1 family. Based on the phylogenetic tree and genetic distanced matrix, the SpLEA1 was found to have high homology with Lea-5 protein from Cicer arietinum and Trifolium pratense . (3) Predictive analysis of cis-acting elements in promoter sequenced using online tools revealed that the SpLEA1 gene promoter contained five classes of hormone response elements and functional elements related to the drought stress response. The SpLEA1 gene was hypothesized to have multiple functions in the plant body and was closely related to drought stress response mechanisms. (4) SpLEA1 gene expression was up-regulated under natural dehydration treatment and peaked at 12 h. After rehydration treatment at 24 h, expression was significantly down-regulated. In summary, the SpLEA1 gene is likely to be involved in the regulation of drought stress response mechanisms in matted curly cypress. This result lays the foundation for further studies on the function of the matted cypress SpLEA1 gene under drought stress and its expression regulation mechanism.
Subjects: Biology >> Botany >> Applied botany submitted time 2020-09-15 Cooperative journals: 《广西植物》
Abstract：为探究纤枝短月藓LEA2 基因的结构和表达特征，该研究以纤枝短月藓为材料，首次利用PCR 克隆技术得到纤枝短月藓BeLEA2 基因序列，对该基因进行分析。结果表明：该基因序列中含有2 个外显子和1 个内含子，其开放阅读框（ORF）为 456 bp，编码151 个氨基酸,预测其相对分子质量为16 515.96 Da。将纤枝短月藓与其他植物LEA2 基因氨基酸序列进行比对，构建系统进化树，结果显示纤枝短月藓与小立碗藓的亲缘关系最近。利用HiTail PCR 技术克隆获得1 072 bp 的BeLEA2 启动子序列，用PlantCARE 在线工具对该启动子的顺式作用元件进行预测，结果表明该启动子除了含有核心启动子元件TATA-box 和CAAT-box 外，还含有ABRE、MYB、MYC、MYB 结合位点（MBS）等其他顺式元件。实时荧光定量PCR 分析表明，BeLEA2 基因在纤枝短月藓不同发育时期和不同组织中都有表达，且对脱水胁迫有响应。以上结果为进一步探究LEA2 基因在苔藓植物中的功能及作用机制奠定了基础。
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