Purification of novel protein elicitor from Botrytis cinerea that induces disease resistance and drought tolerance in plants.
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.
Zhang
Yunhua
Y
Key Laboratory for Biological Control of Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, PR China.
Yang
Xiufen
X
Liu
Quan
Q
Qiu
Dewen
D
Zhang
Yuliang
Y
Zeng
Hongmei
H
Yuan
Jingjing
J
Mao
Jianjun
J
D020171:Botrytis / Q000737:chemistry*; D002852:Chromatography, Ion Exchange; D055864:Droughts; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D007113:Immunity, Innate; D007526:Isoelectric Point; D018551:Solanum lycopersicum / Q000187:drug effects; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010937:Plant Growth Regulators / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D018521:Plant Physiological Phenomena; D010944:Plants / Q000187:drug effects* / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013312:Stress, Physiological; D014908:Triticum / Q000187:drug effects
D005656:Fungal Proteins; D010937:Plant Growth Regulators