Comparative transcriptomic profiling between wild-type and FvMATE51 mutant (a vacuolar membrane protein) strawberry fruits at 26 DPA

To explore the molecular mechanisms of FvMATE51 during fruit development and ripening, we generated CRISPR/Cas9-mediated gene knockout mutants Fvmate51. Then, we performed comparative transcriptome analysis through RNA sequencing (RNA-seq) using fruits of the wild-type and the mutant lines (Fvmate51-1 and Fvmate51-2) at 26 days post anthesis. Three independent biological replicates were applied for the RNA-seq analysis, using Illumina Nova seq 6000 Total RNA was extracted from fruits of 26 DPA WT fruits and FvmMATE1 mutants (Fvmate51-1 and Fvmate51-2) at 26-day postanthesis using HiPure HP Plant RNA Mini Kit (Magen, China).Double-stranded cDNA libraries were prepared from poly(A)+ mRNA, subjected to Illumina high-throughput sequencing after quality control and pooling.

为探究FvMATE51在果实发育与成熟进程中的分子机制，本研究通过CRISPR/Cas9介导的基因敲除技术构建了FvMATE51基因敲除突变体Fvmate51。随后，我们以野生型株系及突变株系Fvmate51-1、Fvmate51-2花后26天的果实为材料，通过RNA测序（RNA-seq）开展比较转录组学分析。本次RNA测序分析设置3次独立生物学重复，采用Illumina NovaSeq 6000平台完成测序。我们使用中国Magen公司的HiPure HP Plant RNA Mini Kit，从花后26天的野生型果实以及FvmMATE1突变体（包含Fvmate51-1与Fvmate51-2株系）的果实中提取总RNA。从poly(A)+ mRNA中制备双链cDNA文库，经质量控制与混合后，开展Illumina高通量测序。