Functional Analysis On StrbohC In Promotion Of Wound Healing In Potato By BTH

The natural healing time of tubers is longer and the mechanism of wound healing is not clear.In this study,the potato tubers‘cv.Longshu No.3'were used as materials.The artificially damaged tubers were treated with BTH at 100mg/L.The weight loss and disease index were determined during wound healing.The differential expression of treated tubers was analyzed on Strbohs family genes.The transgenic seedlings of over-expressed and interfere expressed StrbohC were screened,and the wound effects of transgenic tubers were evaluated.The function of StrbohC of potato tubers during wound healing was clarified.The results showed that as follow:1.BTH treatment promoted wound healing of tubers by reducing the weight loss rate and disease index.BTH promoted the expression of the Strbohs gene during the wound healing of the tuber according to bioinformatics analysis,in which StrbohC up-regulated expression maximum.There was 2902bp cDNA full-length in StrbohC gene and encoded 938 amino acids.In addition,the chromosomal distribution of Strboh family genes was relatively scattered,and the length and number of exons and introns of genes showed different greatly,indicating that each isomer functions differently.2.StrbohC gene was employed to construct overexpression and the artificial miRNA interference expression vectors.After the transformation in Agrobacterium,PCR identification was performed and the corresponding transgenic plants were successfully obtained.By constructing the eGFP fusion protein,the subcellular localization of StrbohC protein was found on the plasma membrane of tobacco epidermal cells.3.The weight loss and disease index of overexpressing StrbohC gene tubers were significantly reduced during wound healing,which were 36.3%and 17%lower than the control on the 7th day after wounded.Overexpressing StrbohC gene tubers also had advanced and more deposition of lignin and suberin at wounds sites.However,the tubers of interference expression of StrbohC gene significantly increased the weight loss and disease index during wound healing,which were 40.6%and 28.7%higher than the control on the 7th day after wounded.In addition,the tubers of interference expression of StrbohC gene had delayed and less at wounds sites,which indicated that StrbohC gene directly involved the structure formation of wound healing.4.The tubers of overexpression StrbohC gene significantly increased the content of H₂O₂ and O₂^-.,the H₂O₂ content was 84.6%higher than the control after 24 hours of wound healing.While the tubers of interference expression of StrbohC gene remarkably reduced the content of H₂O₂ and O₂^-.,which indicated that StrbohC regulated reactive oxygen species accumulation in tubers during wound healing.It is suggested that BTH promotes the wound healing of potato tubers by reducing the weight loss and disease index during wound healing.The family member of Strbohs gene up-regulat during wound healing,StrbohC up-regulates maximum.The evaluation results of tubers with StrbohC overexpression and interfering expression indicate that overexpression tubers have a more effective wounded-healing,and interfering expression ones have a less performance,which indicats that the accumulation of reactive oxygen species involved with StrbohC plays important regulatory functions on wound healing of potato tubers.