Yang Xiaohui, Du Qingzhang, Chen Jinhui, Wang Bowen, Zhang Deqiang
National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry University, No. 35, Qinghua East Road, Beijing, 100083, People's Republic of China,
Planta. 2015 Jul;242(1):77-95. doi: 10.1007/s00425-015-2287-3. Epub 2015 Apr 2.
We used transcript profiling and multi-SNP association to investigate the genetic regulatory relationship between miRNA Pto-miR530a and its target Pto-KNAT1, identifying additive, dominant, and epistatic effects. MicroRNAs (miRNAs) play crucial roles in the post-transcriptional regulation of plant growth and development; indeed, many studies have described the importance of miRNA-target interactions in herbaceous species. However, elucidation of the miRNA-target interactions in trees may require novel strategies. In the present study, we describe a strategy combining expression profiling by reverse transcription quantitative PCR (RT-qPCR) and association mapping with multiple single nucleotide polymorphisms (SNPs) to evaluate the interaction between Pto-miR530a and its target Pto-KNAT1 in Populus tomentosa. RT-qPCR analysis showed a negative correlation (r = -0.62, P < 0.05) between expression levels of Pto-miR530a and Pto-KNAT1 in eight tissues. We used a Bayesian hierarchical model to identify allelic variants of Pto-miR530a and Pto-KNAT1 that associated with eight traits related to growth and wood properties, in a population of 460 unrelated individuals of P. tomentosa. This analysis identified 27 associations, with the proportions of phenotypic variance (R (2)) contributed by each SNP ranging of 0.82-15.81 %, the additive effects of each SNP ranging of 0.16-18.09, and the dominant effects ranging from -14.09 to 19.00. Epistatic interaction models showed a strong interaction among SNPs in the miRNA target with R (2) of 0.1-3.56 %, and information gain of significant SNP pairs of -3.09 to 0.93 %, representing the regulatory interactions between the miRNA and the mRNA. Thus, we used a new strategy that combines association genetics and expression profiling based on SNPs to study the regulatory relationship between this miRNA and its target mRNA, thereby providing novel advances in our understanding of the genetic architecture of important traits.
我们运用转录本分析和多单核苷酸多态性(SNP)关联分析,来研究微小RNA(miRNA)Pto-miR530a与其靶标Pto-KNAT1之间的遗传调控关系,鉴定加性、显性和上位性效应。微小RNA(miRNAs)在植物生长发育的转录后调控中发挥着关键作用;事实上,许多研究都描述了miRNA-靶标相互作用在草本植物中的重要性。然而,阐明树木中的miRNA-靶标相互作用可能需要新的策略。在本研究中,我们描述了一种将逆转录定量PCR(RT-qPCR)表达谱分析与多个单核苷酸多态性(SNPs)关联作图相结合的策略,以评估毛白杨中Pto-miR530a与其靶标Pto-KNAT1之间的相互作用。RT-qPCR分析表明,在八个组织中Pto-miR530a和Pto-KNAT1的表达水平呈负相关(r = -0.62,P < 0.05)。我们使用贝叶斯层次模型,在460个不相关的毛白杨个体群体中,鉴定与八个与生长和木材性质相关性状相关的Pto-miR530a和Pto-KNAT1的等位基因变异。该分析鉴定出27个关联,每个SNP贡献的表型变异比例(R²)范围为0.82 - 15.81%,每个SNP的加性效应范围为0.16 - 18.09,显性效应范围为 - 14.09至19.00。上位性相互作用模型显示,miRNA靶标中的SNP之间存在强烈相互作用,R²为0.1 - 3.56%,显著SNP对的信息增益为 - 3.09至0.93%,代表了miRNA与mRNA之间的调控相互作用。因此,我们采用了一种基于SNP的关联遗传学和表达谱分析相结合的新策略,来研究这种miRNA与其靶标mRNA之间 的调控关系,从而为我们理解重要性状的遗传结构提供了新进展。