Screening of ARF-Aux/IAA Interaction Combinations Involved in Apple Fruit Size
ZHOU Zhe,, BIAN ShuXun, ZHANG HengTao, ZHANG RuiPing, GAO QiMing, LIU ZhenZhen, YAN ZhenLi,Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 459000
Abstract 【Objective】The transcriptomics and bioinformatics methods were used to carry out the genome-wide potential interacting MdARFs and MdIAAs pairs screening, so as to build a foundation for clarifying the function of related genes and elucidating the molecular mechanism underlying auxin-regulated apple fruit size.【Method】 Two apple genotypes, Royal Gala (WT) and 35S::miRNA172p transgenic Royal Gala (miR172OX ) were used as test materials in this study. Fruit materials from different developmental stages and tissue types were collected for both genotypes and were subjected to transcriptome sequencing. Clean reads were aligned to the reference genome and the differential expression analysis was performed. Based on the transcriptome data obtained by sequencing the transgenic small fruit and wild-type large fruit, the pairwise expression analysis was performed across MdARFs and MdIAAs families. The amino acid sequences of 23 ARFs and 34 Aux/IAAs were downloaded from Arabidopsis genome, and 21 ARFs and 25 Aux/IAAs were downloaded from tomato genome, which were further compared with candidate MdARFs and MdIAAs to construction phylogenetic trees. The MEME and TBtools were used to carry out the Motif analysis for candidate MdARFs and MdIAAs. Pairs with high interacting possibilities were further confirmed by a protein-protein interacting network constructed in apple to finalize combinations with the highest probability of involvement in fruit development. 【Result】 The whole fruit at 2 weeks post full bloom and the fruit skin, fruit flesh and fruit core at 4 WPFB were collected from WT and miR172OX, respectively. To achieve research objectives, transcriptome sequencing was carried out. A total of 178.19 Gb paired-end reads of 125 bp/150 bp were generated. All indexes indicated that the three biological replicates had highly consistent transcriptome profiles across all tissue types. FPKM values in at least one library was over 2 were used as a standard to eliminate the low expressed genes, so a total of 38MdARFs and 27 MdIAAs were expressed. In our fruit developmental transcriptome data, eight pairs of MdARF-MdIAA were obtained through Pearson correlation analysis, whose Pearson correlation coefficient was over 0.9 or below -0.9. The systematic phylogenetic analysis showed that MdARF6 and MdARF19 belonged to the same branch with AtARFs, which played a role in transcription activation, while MdARF2, MdARF4, and MdARF9 were closely related to transcriptional inhibitory AtARFs. Motif analysis results showed that both the candidate MdARFs and MdIAAs proteins contained Motif 2 and Motif 5, which were corresponded to the conserved domains Motif IV and Motif III in the IAA protein, respectively. After homolog mapping inspection with Arabidopsis, two potential MdARF-MdIAA interacting pairs were selected for future functional identification. 【Conclusion】Among apple MdARF and MdIAA family members, eight pairs of MdARF-MdIAA showed significant correlations in terms of their expression patterns during fruit development. Further homology mapping confirmed two pairs of them, including MdARF4-MdIAA17 and MdARF4-MdIAA19, were most likely to participate in the regulation of apple fruit development through mediating auxin signal transduction. Keywords:apple;ARF;Aux/IAA;interacting-pair screening
PDF (3387KB)元数据多维度评价相关文章导出EndNote|Ris|Bibtex收藏本文 本文引用格式 周喆, 卞书迅, 张恒涛, 张瑞萍, 高启明, 刘珍珍, 阎振立. 苹果果实大小相关的ARF-Aux/IAA互作组合筛选[J]. 中国农业科学, 2021, 54(14): 3088-3096 doi:10.3864/j.issn.0578-1752.2021.14.014 ZHOU Zhe, BIAN ShuXun, ZHANG HengTao, ZHANG RuiPing, GAO QiMing, LIU ZhenZhen, YAN ZhenLi. Screening of ARF-Aux/IAA Interaction Combinations Involved in Apple Fruit Size[J]. Scientia Acricultura Sinica, 2021, 54(14): 3088-3096 doi:10.3864/j.issn.0578-1752.2021.14.014
A:热图显示苹果ARF和IAA基因家族表达量的聚类分析;B:苹果ARF和IAA基因家族中的差异表达基因及差异表达趋势 Fig. 2Expression analysis of MdARF and MdIAA encoding genes
A: Transcriptional heat maps showing expression levels of apple ARF and IAA encoding genes; B: Differentially expressed genes in apple ARF and IAA gene families and their differential expression trends
A:苹果候选ARF-Aux/IAA互作对中MdARFs和MdIAAs的系统发育分析;B:苹果候选ARF-Aux/IAA互作对中MdARFs和MdIAAs的保守结构域分析 Fig. 4The phylogenetic tree and conserved motif analysis of interacting candidate combination members from ARF and IAA gene families in apple
A: Phylogenetic analysis of MdARFs and MdIAAs in apple candidate ARF-Aux/IAA interaction pairs; B: Conserved domain analysis of MdARFs and MdIAAs in apple candidate ARF-Aux/IAA interaction pairs
节点大小对应节点连接数量,节点颜色对应Log2(Foldchange)值,节点间连线颜色对应PCC值 Fig. 5Putative Protein-Protein Interaction network involving in MdARFs and MdIAAs in apple
Node sizes correspond to the degree of connectivity of the nodes, nodes are color-coded according to the Log2 (Foldchange) values and edge are color-coded according to the PCC values
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