Purification of novel protein elicitor from Botrytis cinerea that induces disease resistance and drought tolerance in plants

Abstract

PebC1, a novel protein elicitor was isolated and purified from the mycelium of gray mold fungus, Botrytis cinerea strain BC-4-2-2-1. The protein was eluted through HiTrap DEAE FF and RESOURCE Q anion exchange chromatography and displayed as a single band with an apparent molecular weight of 36 kDa on silver staining sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The pI of the purified protein PebC1 was determined by 2-DE and was 4.85. Three peptide segments were obtained by MALDI-TOF. Similarity, the homology matching using protein BLAST search found that two proteins, viz. XP_001593856 and XP_001551609 were having high score and covered sequence of the three peptides. Protein XP_001551609, a deduced protein nascent polypeptide-associated complex alpha-polypeptide, was more authentic because it was from Botryotinia fuckeliana that is better known as its anamorph, B.cinerea and showed 95% homology with the three polypeptides. The full cDNA sequence encoding for pebC1 (Genbank accession number FJ748868) was amplified from B. cinerea and consists of 639bp, which is same as a registered gene of XM_001551559, a nascent polypeptide-associated complex alpha-polypeptide partial mRNA. The gene encode a hypothetical protein speculated from an annotated genomic sequence from B. fuckeliana B05.10 (NW_001814507) and there is no publication about the gene. The PebC1 protein significantly promoted wheat seedling growth with an optimum protein concentration of 5 microg/mL. Root systemic activity of wheat with 4-5 leaves increased by 1.29 fold, and the wheat seedling drought resistance integrated index increased from 36.53 to 57.08 under two cycles of drought stress after treatment of PebC1. PebC1 protein at the optimum concentration of 10 microg/mL induced 69.19% disease resistance against gray mold fungus in tomato. Furthermore, phenylalanine ammonia-lyase (PAL), peroxides (POD), and polyphenol oxidase (PPO) related to plant resistance metabolism were also increased considerably after PebC1 treatment. PAL activity was increased by 46.84% at 24h post-treatment, while POD and PPO activity increased by 109.5% and 111.0% at 72 h, respectively over the control.

Non-specific lipid transfer proteins (ns-LTPs) from maize induce resistance in pearl millet against downy mildew disease

“…The figures specify that the level of POX activity remained high at 48 hpi after which the enzyme activity gradually dropped in both inoculated and uninoculated samples. In the same way increase in POX activity was reported in tomato plants treated with Peb C1 protein elicitor from B. Cinerea (Zhang et al 2010). In our study, there was increase in POX activity in the maize ns-LTP M1 treated seedlings upon challenge inoculation with the pathogen compared to uninoculated susceptible seedlings.…”

“…A protein elicitor isolated from S. graminicola induced activation of phenyl alanine ammonia lyase (PAL) and peroxidase (POX) activity (Sharathchandra et al 2006) and homologous proteins like INF1, NPP1 (Baillieul et al 2003), cryptogein, PcF (Orsomando et al 2003), PB90 (Wang et al 2003), megaspermin and quercinin (Koehl et al 2003) with the similar biological activity have been found in other Phytophthora spp., an oomycete plant pathogen. The protein elicitor PebC1 from Botrytis cinerea strain BC-4-2-2-1 promoted seedling growth, drought tolerance in wheat and also induce disease resistance against gray mold fungus in tomato (Zhang et al 2010).…”

A novel elicitor identified from Magnaporthe oryzae triggers defense responses in tobacco and rice

“…As observed for previously reported elicitors, such as MoHrip1 (Chen et al 2012), PevD1 (Wang et al 2012), PebC1 (Zhang et al 2009), andHrip1 (Kulye et al 2012), the elicitor protein MoHrip2 can trigger the accumulation of early signaling ROS in tobacco cells and enhance the resistance of rice seedlings against infection by M. oryzae. Moreover, MoHrip2 also triggered the accumulation of NO, which is another important early signaling molecule, in tobacco cells.…”

PeBL1, a Novel Protein Elicitor from Brevibacillus laterosporus Strain A60, Activates Defense Responses and Systemic Resistance in Nicotiana benthamiana

“…The results of the mass spectrometry analysis were searched by Mascot. Based on the Mascot search results, we obtained the best-matching protein, which had the highest score (194) and contained two different reliable peptides: (i) 45 TSNETWNLGSHIR 57 (score, 67) and (ii) 88 FTAVQPGNASIYVYK 102 (score, 47). MOWSE scores greater than 25 were significant for these search results.…”

“…The signs and diameters of TMV lesions in each leaf were analyzed at 2, 4, and 6 days postinoculation (p.i. ), as previously described (44,45). For the diameters of the lesions, 10 random lesions were measured for each plant.…”