A novel polypeptide named as defensin CM was isolated from Chinese mistletoe, Viscum coloratura （Kom.） Nakai. The amino acid sequence was determined by the combination of Edman degradation, endoproteinase Lys-C dige...A novel polypeptide named as defensin CM was isolated from Chinese mistletoe, Viscum coloratura （Kom.） Nakai. The amino acid sequence was determined by the combination of Edman degradation, endoproteinase Lys-C digestion, and MALDI-TOF mass spectrometry. The primary structure was ATCSAPSGRF KGACFSSNTC SNICKTLEGL KDGHCTGLAC YCSRNC.展开更多
Gerbera(Gerbera hybrida),a major fresh cut flower crop,is very susceptible to root rot disease.Although plant defensins(PDFs),a major group of plant antimicrobial peptides,display broad-spectrum antifungal and antibac...Gerbera(Gerbera hybrida),a major fresh cut flower crop,is very susceptible to root rot disease.Although plant defensins(PDFs),a major group of plant antimicrobial peptides,display broad-spectrum antifungal and antibacterial activities,PDF genes in gerbera have not been systematically characterized.Here,we identified and cloned nine PDF genes from gerbera and divided them into two classes based on phylogenetic analysis.Most Class Ⅰ GhPDF genes were highly expressed in petioles,whereas all Class Ⅱ GhPDF genes were highly expressed in roots.Phytophthora cryptogea inoculation strongly upregulated all Class II GhPDF genes in roots and upregulated all Class Ⅰ GhPDF genes in petioles.Transient overexpression of GhPDF1.5 and GhPDF2.4 inhibited P.cryptogea infection in tobacco(Nicotiana benthamiana)leaves.Transient overexpression of GhPDF2.4,but not GhPDF1.5,significantly upregulated ACO and LOX gene expression in tobacco leaves,indicating that overexpressing GhPDF2.4 activated the jasmonic acid/ethylene defense pathway and that the two types of GhPDFs have different modes of action.Prokaryotically expressed recombinant GhPDF2.4 inhibited mycelial growth and delayed the hyphal swelling of P.cryptogea,in vitro,indicating that GhPDF2.4 is a morphogenetic defensin.Moreover,the addition of GhPDF2.4 to plant culture medium alleviated the root rot symptoms of in vitro-grown gerbera seedlings and greatly reduced pathogen titer in P.cryptogea-inoculated gerbera roots in the early stages of treatment.Our study provides a basis for the use of GhPDFs,especially GhPDF2.4,for controlling root rot disease in gerbera.展开更多
Monocot high expression vector pBARUNP1, harboring rabbit defensin(NP 1) gene and selective bar gene for resistance to the herbicide Basta, were constructed and then transferred into immature embryos of wheat (“Bobw...Monocot high expression vector pBARUNP1, harboring rabbit defensin(NP 1) gene and selective bar gene for resistance to the herbicide Basta, were constructed and then transferred into immature embryos of wheat (“Bobwhite” and “Zhong 60634”)via particle bombardment. Southern and RNA dot blots showed the stable integration and transcription of foreign NP 1 gene in the wheat genome. Furthermore, in vitro microbicidal activity assay indicated the proper translation of defensin. Crude protein extraction of transgenic plants exhibited to some extent cytotoxic to several pathogens including G. saubinetii, B. subtilis, E.coli, and A. tumefaciens.展开更多
文摘A novel polypeptide named as defensin CM was isolated from Chinese mistletoe, Viscum coloratura （Kom.） Nakai. The amino acid sequence was determined by the combination of Edman degradation, endoproteinase Lys-C digestion, and MALDI-TOF mass spectrometry. The primary structure was ATCSAPSGRF KGACFSSNTC SNICKTLEGL KDGHCTGLAC YCSRNC.
基金supported by the Construction Project for Shanxi Key Laboratory of Germplasm Resources Innovation and Utilization of Vegetables and Flowers(202204010931019)the Reward Fund for PhDs and Postdoctors of Shanxi Province(SXBYKY2022004)the Fund for High-level Talents of Shanxi Agricultural University(2021XG010).
文摘Gerbera(Gerbera hybrida),a major fresh cut flower crop,is very susceptible to root rot disease.Although plant defensins(PDFs),a major group of plant antimicrobial peptides,display broad-spectrum antifungal and antibacterial activities,PDF genes in gerbera have not been systematically characterized.Here,we identified and cloned nine PDF genes from gerbera and divided them into two classes based on phylogenetic analysis.Most Class Ⅰ GhPDF genes were highly expressed in petioles,whereas all Class Ⅱ GhPDF genes were highly expressed in roots.Phytophthora cryptogea inoculation strongly upregulated all Class II GhPDF genes in roots and upregulated all Class Ⅰ GhPDF genes in petioles.Transient overexpression of GhPDF1.5 and GhPDF2.4 inhibited P.cryptogea infection in tobacco(Nicotiana benthamiana)leaves.Transient overexpression of GhPDF2.4,but not GhPDF1.5,significantly upregulated ACO and LOX gene expression in tobacco leaves,indicating that overexpressing GhPDF2.4 activated the jasmonic acid/ethylene defense pathway and that the two types of GhPDFs have different modes of action.Prokaryotically expressed recombinant GhPDF2.4 inhibited mycelial growth and delayed the hyphal swelling of P.cryptogea,in vitro,indicating that GhPDF2.4 is a morphogenetic defensin.Moreover,the addition of GhPDF2.4 to plant culture medium alleviated the root rot symptoms of in vitro-grown gerbera seedlings and greatly reduced pathogen titer in P.cryptogea-inoculated gerbera roots in the early stages of treatment.Our study provides a basis for the use of GhPDFs,especially GhPDF2.4,for controlling root rot disease in gerbera.
文摘Monocot high expression vector pBARUNP1, harboring rabbit defensin(NP 1) gene and selective bar gene for resistance to the herbicide Basta, were constructed and then transferred into immature embryos of wheat (“Bobwhite” and “Zhong 60634”)via particle bombardment. Southern and RNA dot blots showed the stable integration and transcription of foreign NP 1 gene in the wheat genome. Furthermore, in vitro microbicidal activity assay indicated the proper translation of defensin. Crude protein extraction of transgenic plants exhibited to some extent cytotoxic to several pathogens including G. saubinetii, B. subtilis, E.coli, and A. tumefaciens.
