Function identification of MdTIR1 in apple root growth benefited from the predicted MdPPI network
Function identification of MdTIR1 in apple root growth benefited from the predicted MdPPI network
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摘要:
Protein–protein interaction (PPI) network anal-ysis is an effective method to identify key pro-teins during plant development, especially in species for which basic molecular research is lacking, such as apple (Malus domestica). Here, an MdPPI network containing 30806 PPIs was inferred in apple and its quality and reliability were rigorously verified. Subsequently, a root-growth subnetwork was extracted to screen for critical proteins in root growth. Because hormone-related proteins occupied the largest proportion of critical proteins, a hormone-related sub-subnetwork was further extracted from the root-growth subnetwork. Among these proteins, auxin-related M. domestica TRANS-PORT INHIBITOR RESISTANT 1 (MdTIR1) served as the central, high-degree node, implying that this protein exerts essential roles in root growth. Furthermore, transgenic apple roots over-expressing an MdTIR1 transgene displayed in-creased primary root elongation. Expression analysis showed that MdTIR1 significantly up-regulated auxin-responsive genes in apple roots, indicating that it mediates root growth in an auxin-dependent manner. Further experimental validation revealed that MdTIR1 interacted with and accelerated the degradation of MdIAA28, MdIAA43, and MdIAA46. Thus, MdTIR1-mediated degradation of MdIAAs is critical in auxin signal transduction and root growth regu-lation in apple. Moreover, our network analysis and high-degree node screening provide a novel research technique for more generally charac-terizing molecular mechanisms.