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枯草芽孢杆菌B25的一种新型抗真菌蛋白。

A novel antifungal protein of Bacillus subtilis B25.

作者信息

Tan Zhiqiong, Lin Baoying, Zhang Rongyi

机构信息

Key Laboratory of Protection and Development Utilization of Tropical Crop Germplasm Resources, Hainan University, 58#, Renmin road, 570228 Haikou, Hainan Province China.

出版信息

Springerplus. 2013 Oct 17;2:543. doi: 10.1186/2193-1801-2-543. eCollection 2013.

DOI:10.1186/2193-1801-2-543
PMID:24255843
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3824700/
Abstract

Bacillus subtilis B25 was isolated from banana rhizosphere soil. It has been confirmed for B25 to have stronger antagonism against Fusarium oxysporum f.sp.cubense, Additionally B25 has good inhibitory to plant pathogens, including Corynespora cassiicola, Alternaria solani, Botrytis cinerea and Colletotrichum gloeosporioides on potato dextrose agar (PDA) plates. The antagonistic substance can be extracted from cell-free culture broth supernatants by 70% (w/v) (NH4)2 SO4 saturation. Clear blank band was observed between the protein and a pathogen. The examination of antagonistic mechanism under light microscope showed that the antifungal protein of B25 appeared to inhibit pathogens by leading to mycelium and spores tumescence, distortion, abnormality. The isolation procedure comprised ion exchange chromatography on DEAE-Sephadex Fast Flow and gel filtration chromatography on SephadexG-100. The purified antifungal fraction showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The active fraction was identified by NanoLC-ESI-MS/MS The amino acid sequences of 17 peptides segments were obtained. The analysis of the protein suggested that it was a hypothetical protein (gi154685475), with a relative molecular mass of 38708.67 Da and isoelectric point (pI) of 5.63.

摘要

枯草芽孢杆菌B25是从香蕉根际土壤中分离得到的。已证实B25对香蕉枯萎病菌具有较强的拮抗作用。此外，B25对植物病原菌具有良好的抑制作用，在马铃薯葡萄糖琼脂（PDA）平板上，其对包括瓜类炭疽病菌、番茄早疫病菌、灰葡萄孢菌和胶孢炭疽菌在内的植物病原菌均有抑制作用。拮抗物质可通过70%（w/v）硫酸铵饱和度从无细胞培养液上清中提取。在蛋白质和病原菌之间观察到清晰的空白带。光学显微镜下对拮抗机制的检查表明，B25的抗真菌蛋白似乎通过导致菌丝体和孢子肿胀、变形、异常来抑制病原菌。分离过程包括在DEAE-葡聚糖快速流动柱上进行离子交换色谱和在SephadexG-100柱上进行凝胶过滤色谱。纯化的抗真菌组分在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）中显示为单一条带。通过纳升液相色谱-电喷雾串联质谱（NanoLC-ESI-MS/MS）对活性组分进行鉴定，获得了17个肽段的氨基酸序列。蛋白质分析表明它是一种假定蛋白（gi154685475），相对分子质量为38708.67 Da，等电点（pI）为5.63。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/5be9de6a3bae/40064_2013_630_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/f85581c6d4ce/40064_2013_630_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/4d2e42e4c8b4/40064_2013_630_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/b067c9a6c6f9/40064_2013_630_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/eba44fc124b1/40064_2013_630_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/abaa83dc5522/40064_2013_630_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/5be9de6a3bae/40064_2013_630_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/f85581c6d4ce/40064_2013_630_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/4d2e42e4c8b4/40064_2013_630_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/b067c9a6c6f9/40064_2013_630_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/eba44fc124b1/40064_2013_630_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/abaa83dc5522/40064_2013_630_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1298/3824700/5be9de6a3bae/40064_2013_630_Fig6_HTML.jpg

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