目的 对可能参与花青素代谢调控的黑果枸杞R1-MYB转录因子基因进行克隆、生物信息学分析和不同品种、同一品种不同器官及盐胁迫条件下差异表达分析。方法 通过同源基因克隆和RACE方法得到黑果枸杞R1-MYB转录因子编码区全长，通过转录组数据获得宁夏枸杞同源基因序列。通过Prot、Param、Smart、PSORT和SOPMA进行生物信息学分析，通过MEGA 5.0构建NJ系统进化树。同时采用Real-time PCR的方法进行基因表达分析。结果 获得黑果枸杞LrMYB1R1（KY568981）及宁夏枸杞LbMYB1R1（KY568982）基因全长cDNA，cDNA编码区全长1 496 bp，编码区为927 bp，编码产物包含308个氨基酸，编码蛋白相对分子质量分别为33 400和33 490，理论等电点分别为7.80和7.78，属于R1-MYB转录因子，经预测其编码蛋白位于细胞核中；LrMYB1R1和LbMYB1R1与茄科植物番茄、马铃薯、烟草中的MYB1R1-like蛋白具有高度相似性。LrMYB1R1的表达表现出器官和发育阶段中差异性，具有品种特异性，LrMYB1R1的表达受盐胁迫抑制。结论 丰富了对R1-MYB转录因子的研究，为接下来的基因功能研究及利用基因工程手段提高黑果枸杞中花青素含量奠定基础。

Objective To clone the R1-MYB transcription factor participated in the anthocyanidin metabolism, and to analyze by bioinformatics analysis. Different expression of different varieties, different organs of the same species and salt stress conditions in Lycium were analyzed. To clone the full-length cDNA encoding R1-MYB, to perform bioinformatic analysis, and to study its expression in different cultivators and different developmental stage and in response to NaCl stress in Lycium ruthenicum and L. barbarum. Methods The full-length cDNA encoding R1-MYB was cloned using homology-based cloning and rapid amplification of cDNA ends (RACE) technique in L. ruthenicum, and the homologous gene was obtained by transcriptome in L. barbarum. The bioinformatics analysis was carried out by using Prot, Param, Smart, PSORT, and SOPMA methods. And the phylogenetic tree was constructed based on software MEGA5.0. Gene expression analysis was done by method of Real-time PCR. Results We the MYB transcription factor in L. ruthenicum was cloned and named as LrMYB1R1 (GenBank accession number KY568981), and LbMYB1R1 (GenBank accession number KY568982) in L. barbarum. Bioinformatics analysis showed that the length of LrMYB1R1 was 1 496 bp and the CDS was 927 bp. The coding products contained 308 amino acids, the molecular weight of the protein was 33 400 and 33 490, the theoretical isoelectric point was 7.80 and 7.78, belonging to the R1-MYB transcription factor, and the encoded protein is predicted to be located in the nucleus. The results of phylogenetic tree analysis showed that LrMYB1R1 and LbMYB1R1 were highly similar to MYB1R1-like protein in Solanum lycopersicum, Solanum tuberosum, and Nicotiana tabacum. Real-time PCR analysis showed that LrMYB1R1 had higher expression level in leaves and young fruits in L. ruthenicum, followed by stems, young leaves, flowers, purple fruits and black fruits, only slightly expressed in roots. In addition, the relative expression levels of LrMYB1R1 decreased in response to salt stress. Conclusion The study of R1 MYB transcription factor has been enriched, which has laid the foundation for the subsequent research on gene function and for the high-yielding anthocyanin by genetic engineering method in L. ruthenicum.

宁夏自然科学基金资助项目（NZ16215）