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Antimicrobial peptide tobacco

Xiu-Mei Luo, Cheng-Jian Xie, De Wang, Yun-Min Wei, Jie Cai, Shan-Shan Cheng, Xing -Yong Yang, An -Ping Sui
Psc-AFP, isolated from the seeds of Psoralea corylifolia L., is an antimicrobial protein with trypsin inhibitor activity. Its encoding gene was cloned by 3'- rapid amplification of cDNA ends (RACE) combined with Y-shaped adaptor-dependent extension (YADE) method. The gene Psc-AFP encodes a protein of 203 amino acids with a deduced signal peptide of 24 residues. The growth inhibition effect exerted by the heterologously expressed Psc-AFP in Pichia pastoris revealed that the recombinant Psc-AFP inhibited mycelium growth of Aspergillus niger, Rhizoctonia solani, and Alternaria brassicae and conidial germination of Alternaria alternata...
September 2, 2016: Applied Microbiology and Biotechnology
Roman A Komakhin, Denis A Vysotskii, Rahim R Shukurov, Vera D Voblikova, Vera V Komakhina, Svetlana R Strelnikova, Ekaterina M Vetchinkina, Alexey V Babakov
BACKGROUND: In a previous study we found that in chickweed the expression level of the pro-SmAMP2 gene was comparable or even higher to that of the β-actin gene. This high level of the gene expression has attracted our attention as an opportunity for the identification of novel strong promoters of plant origin, which could find its application in plant biotechnology. Therefore, in the present study we focused on the nucleotide sequence identification and the functional characteristics of the pro-SmAMP2 promoter in transgenic plants...
2016: BMC Biotechnology
Sílvia Vilà, Esther Badosa, Emilio Montesinos, Marta Planas, Lidia Feliu
Cyclolipopeptides derived from the antimicrobial peptide c(Lys-Lys-Leu-Lys-Lys-Phe-Lys-Lys-Leu-Gln) (BPC194) were prepared on solid-phase and screened against four plant pathogens. The incorporation at Lys5 of fatty acids of 4 to 9 carbon atoms led to active cyclolipopeptides. The influence on the antimicrobial activity of the Lys residue that is derivatized was also evaluated. In general, acylation of Lys1, Lys2 or Lys5 rendered the sequences with the highest activity. Incorporation of a D-amino acid maintained the antimicrobial activity while significantly reduced the hemolysis...
2016: PloS One
Qasim Al Souhail, Yasuaki Hiromasa, Mohammad Rahnamaeian, Martha C Giraldo, Daisuke Takahashi, Barbara Valent, Andreas Vilcinskas, Michael R Kanost
Insects secrete antimicrobial peptides as part of the innate immune response. Most antimicrobial peptides from insects have antibacterial but not antifungal activity. We have characterized an antifungal peptide, diapausin-1 from hemolymph of a lepidopteran insect, Manduca sexta (tobacco hornworm). Diapausin-1 was isolated by size exclusion chromatography from hemolymph plasma of larvae that were previously injected with a yeast, Saccharomyces cerevisiae. Fractions containing activity against S. cerevisiae were analyzed by SDS-PAGE and MALDI-TOF MS/MS and found to contain a 45-residue peptide that was encoded by sequences identified in M...
August 2016: Developmental and Comparative Immunology
Xue Zhong, Xiang-Jun Rao, Hui-Yu Yi, Xin-Yu Lin, Xiao-Hong Huang, Xiao-Qiang Yu
Nuclear factor κB (NF-κB) plays an essential role in regulation of innate immunity. In mammals, NF-κB factors can form homodimers and heterodimers to activate gene expression. In insects, three NF-κB factors, Dorsal, Dif and Relish, have been identified to activate antimicrobial peptide (AMP) gene expression. However, it is not clear whether Dorsal (or Dif) and Relish can form heterodimers. Here we report the identification and functional analysis of a Dorsal homologue (MsDorsal) and two Relish short isoforms (MsRel2A and MsRel2B) from the tobacco hornworm, Manduca sexta...
2016: Scientific Reports
Yun-Peng Wang, Zheng-Yi Wei, Yu-Ying Zhang, Chun-Jing Lin, Xiao-Fang Zhong, Yue-Lin Wang, Jing-Yong Ma, Jian Ma, Shao-Chen Xing
Rice blast is a major destructive fungal disease that poses a serious threat to rice production and the improvement of blast resistance is critical to rice breeding. The antimicrobial peptide MSI-99 has been suggested as an antimicrobial peptide conferring resistance to bacterial and fungal diseases. Here, a vector harboring the MSI-99 gene was constructed and introduced into the tobacco chloroplast genome via particle bombardment. Transformed plants were obtained and verified to be homoplastomic by PCR and Southern hybridization...
2015: International Journal of Molecular Sciences
Kimberly Booth, Lizzette Cambron, Nathan Fisher, Kendra J Greenlee
Research on how insect immunity changes with age as insects develop within an instar, or larval developmental stage, is limited and contradictory. Insects within an instar are preparing for the next developmental stage, which may involve changes in morphology or habitat. Immunity may also vary accordingly. To determine how immunity varies in the fifth instar, we tested humoral immune responses, antimicrobial peptide activity, and phenoloxidase activity using the tobacco hornworm, Manduca sexta. We determined that while M...
March 2015: Physiological and Biochemical Zoology: PBZ
Irshad Ahmad Rather, Praveen Awasthi, Vidushi Mahajan, Yashbir S Bedi, Ram A Vishwakarma, Sumit G Gandhi
Pathogenesis-related (PR) proteins are involved in biotic and abiotic stress responses of plants and are grouped into 17 families (PR-1 to PR-17). PR-5 family includes proteins related to thaumatin and osmotin, with several members possessing antimicrobial properties. In this study, a PR-5 gene showing a high degree of homology with osmotin-like protein was isolated from sweet basil (Ocimum basilicum L.). A complete open reading frame consisting of 675 nucleotides, coding for a precursor protein, was obtained by PCR amplification...
March 1, 2015: Gene
Sunita Patro, Soumitra Maiti, Santosh Kumar Panda, Nrisingha Dey
We describe the use of plant-made β-defensins as effective antimicrobial substances for controlling salmonellosis, a deadly infection caused by Salmonella typhimurium (referred to further as S. typhi). Human β-defensin-1 (hBD-1) and -2 (hBD-2) were expressed under the control of strong constitutive promoters in tobacco plants, and bio-active β-defensins were successfully extracted. In the in vitro studies, enriched recombinant plant-derived human β-defensin-1 (phBD-1) and -2 (phBD-2) obtained from both T1 and T2 transgenic plants showed significant antimicrobial activity against Escherichia coli and S...
April 2015: Transgenic Research
Dipak K Sahoo, Sumita Raha, James T Hall, Indu B Maiti
To enhance the natural plant resistance and to evaluate the antimicrobial properties of phylloplanin against blue mold, we have expressed a synthetic chimeric native-phylloplanin-GFP protein fusion in transgenic Nicotiana tabacum cv. KY14, a cultivar that is highly susceptible to infection by Peronospora tabacina. The coding sequence of the tobacco phylloplanin gene along with its native signal peptide was fused with GFP at the carboxy terminus. The synthetic chimeric gene (native-phylloplanin-GFP) was placed between the modified Mirabilis mosaic virus full-length transcript promoter with duplicated enhancer domains and the terminator sequence from the rbcSE9 gene...
2014: TheScientificWorldJournal
Eric T Johnson, Patrick F Dowd, Stephen R Hughes
Lycotoxin I, from the wolf spider (Lycosa carolinensis), is an amphipathic pore-forming peptide that has antimicrobial and anti-insect activity. Constitutive expression of a lycotoxin I modified for oral toxicity to insects in tobacco (Nicotiana tabacum) conferred significantly enhanced resistance to larvae of the corn earworm (Helicoverpa zea) and cigarette beetle (Lasioderma serricorne). Gene expression levels of modified lycotoxin I were negatively correlated to the survival of corn earworm larvae. In addition, pathogenic symptoms caused by Pseudomonas syringae pathovar tabaci and Alternaria alternata on the modified lycotoxin I-expressing leaves were significantly less severe than on wild type leaves...
August 2014: Biotechnology Letters
Shuguang Zhang, Xiaolong Cao, Yan He, Steve Hartson, Haobo Jiang
The tobacco hornworm, Manduca sexta, has been used as a biochemical model for studying insect physiological processes. While the transcriptomes of its fat body, hemocytes, midgut, and antennae have been examined in several studies, limited information is available for proteins in tissues, cells, or body fluids of this insect. In keeping pace with the M. sexta genome project, we launched a pilot study to identify differences in the peptidome of cell-free hemolymph samples from larvae injected with buffer or a mixture of bacteria...
April 2014: Insect Biochemistry and Molecular Biology
Xiufeng Zhang, Yun Zheng, Guru Jagadeeswaran, Ren Ren, Ramanjulu Sunkar, Haobo Jiang
The tobacco hornworm Manduca sexta has served as a model for insect biochemical and physiological research for decades. However, knowledge of the posttranscriptional regulation of gene expression by microRNAs is still rudimentary in this species. Our previous study (Zhang et al., 2012) identified 163 conserved and 13 novel microRNAs in M. sexta, most of which were present at low levels in pupae. To identify additional M. sexta microRNAs and more importantly to examine their possible roles in the expression regulation of immunity-related genes, we constructed four small RNA libraries using fat body and hemocytes from naïve or bacteria-injected larvae and obtained 32...
April 2014: Insect Biochemistry and Molecular Biology
Yan Li, Jiarong Wang, Jing Yang, Chanjuan Wan, Xiaoming Wang, Hongbin Sun
ORBK (LKGCWTKSIPPKPCFK) is a cyclic cationic peptide that has potent antimicrobial properties and trypsin inhibitory activities. To explore a new approach for expressing ORBK in Escherichia coli, a sequence encoding ORBK was cloned into pET28a vector in which maltose-binding protein (MBP) was used as a fusion partner and an N-terminal 6-His as an affinity tag. Protein expression was induced with 0.5mM Isopropyl-thio-galactoside (IPTG) for 4h at 37°C. The recombinant ORBK was then purified by Ni affinity column and further digested with tobacco etch virus (TEV) enzyme...
March 2014: Protein Expression and Purification
YongJoong Kim, Hag Dong Kim, Buhyun Youn, Yun Gyu Park, Joon Kim
Protein secretion is a general phenomenon by which cells communicate with the extracellular environment. Secretory proteins, including hormones, enzymes, toxins, and antimicrobial peptides have various functions in extracellular environments. Here, we determined that ribosomal protein S3 (rpS3) is homodimerized and secreted in several cancer cell lines such as HT1080 (human fibrosarcoma) and MPC11 (mouse plasmacytoma). Moreover, we found that the secreted rpS3 protein increased in doxorubicin-resistant MPC11 cells compared to that in MPC11 cells...
November 29, 2013: Biochemical and Biophysical Research Communications
S El Quakfaoui, C Potvin, R Brzezinski, A Asselin
Growth inhibition towards Rhizopus nigricans, Fusarium oxysporum f. sp. radicis-lycopersici, Verticillium albo-atrum and Pythium ultimum was observed in vitro using a purified chitosanase from an actinomycete, Streptomyces sp, strain N174. The corresponding gene, with its own signal peptide, was inserted into pBI121.7 shuttle vector to transform tobacco. Transgenic plants were analysed for chitosanase activity by a sodium dodecyl sulfate-polyacrylamide gel electrophoresis assay. Two major and one minor active electrophoretic forms were detected in transgenic tobacco...
December 1995: Plant Cell Reports
Nuri Company, Anna Nadal, José-Luis La Paz, Sílvia Martínez, Stefan Rasche, Stefan Schillberg, Emilio Montesinos, Maria Pla
Synthetic linear antimicrobial peptides with cationic α-helical structures, such as BP100, are valuable as novel therapeutics and preservatives. However, they tend to be toxic when expressed at high levels as recombinant peptides in plants, and they can be difficult to detect and isolate from complex plant tissues because they are strongly cationic and display low extinction coefficient and extremely limited immunogenicity. We therefore expressed BP100 with a C-terminal tag which preserved its antimicrobial activity and demonstrated significant accumulation in plant cells...
January 2014: Plant Biotechnology Journal
Arne Weinhold, Mario Kallenbach, Ian Thomas Baldwin
BACKGROUND: Genetically modified plants are widely used in agriculture and increasingly in ecological research to enable the selective manipulation of plant traits in the field. Despite their broad usage, many aspects of unwanted transgene silencing throughout plant development are still poorly understood. A transgene can be epigenetically silenced by a process called RNA directed DNA methylation (RdDM), which can be seen as a heritable loss of gene expression. The spontaneous nature of transgene silencing has been widely reported, but patterns of acquirement remain still unclear...
2013: BMC Plant Biology
Kwang-Chul Kwon, Dheeraj Verma, Shuangxia Jin, Nameirakpam D Singh, Henry Daniell
Plastids sustain life on this planet by providing food, feed, essential biomolecules and oxygen. Such diverse metabolic and biosynthetic functions require efficient communication between plastids and the nucleus. However, specific factors, especially large molecules, released from plastids that regulate nuclear genes have not yet been fully elucidated. When tobacco and lettuce transplastomic plants expressing GFP within chloroplasts, were challenged with Erwinia carotovora (biotic stress) or paraquat (abiotic stress), GFP was released into the cytoplasm...
2013: PloS One
Brigitte M E Hayes, Mark R Bleackley, Jennifer L Wiltshire, Marilyn A Anderson, Ana Traven, Nicole L van der Weerden
In recent decades, pathogenic fungi have become a serious threat to human health, leading to major efforts aimed at characterizing new agents for improved treatments. Promising in this context are antimicrobial peptides produced by animals and plants as part of innate immune systems. Here, we describe an antifungal defensin, NaD1, with activity against the major human pathogen Candida albicans, characterize the mechanism of killing, and identify protection strategies used by the fungus to survive defensin treatment...
August 2013: Antimicrobial Agents and Chemotherapy
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