Phylogenetic Analysis of the Membrane Attack Complex/Perforin Domain-Containing Proteins in Gossypium and the Role of GhMACPF26 in Cotton Under Cold Stress

Abstract

The membrane attack complex/perforin (MACPF) domain-containing proteins are involved in the various developmental processes and in responding to diverse abiotic stress. The function and regulatory network of the <i>MACPF</i> genes are rarely reported in <i>Gossypium</i> spp. We study the detailed identification and partial functional verification of the members of the MACPF family. Totally, 100 putative MACPF proteins containing complete MACPF domain were identified from the four cotton species. They were classified into three phylogenetic groups and underwent multifold pressure indicating that selection produced new functional differentiation. Cotton <i>MACPF</i> gene family members expanded mainly through the whole-genome duplication (WGD)/segmental followed by the dispersed. Expression and <i>cis-</i>acting elements analysis revealed that MACPFs play a role in resistance to abiotic stresses, and some selected <i>GhMACPFs</i> were able to respond to the PEG and cold stresses. Co-expression analysis showed that <i>GhMACPFs</i> might interact with valine-glutamine (VQ), WRKY, and Apetala 2 (AP2)/ethylene responsive factor (ERF) domain-containing genes under cold stress. In addition, silencing endogenous <i>GhMACPF26</i> in cotton by the virus-induced gene silencing (VIGS) method indicated that <i>GhMACPF26</i> negatively regulates cold tolerance. Our data provided a comprehensive phylogenetic evolutionary view of <i>Gossypium</i> MACPFs. The MACPFs may work together with multiple transcriptional factors and play roles in acclimation to abiotic stress, especially cold stress in cotton.