MALDI-TOF-MS技术在大型食用菌甄别鉴定中的应用

目的通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术,对松茸、姬松茸、金耳、银耳、竹荪、榛蘑等常见食用菌进行蛋白图谱分析,探索MALDI-TOF-MS在大型食用菌鉴定领域的应用方法。方法通过甲酸提取法、液氮甲酸法、甲酸超声破碎法、液氮甲酸超声破碎法4种前处理方法分别获得菌菇样本的蛋白提取液;经MALDI-TOF-MS采集样本指纹图谱,对不同方法及不同样本的指纹图谱进行对比分析与聚类分析。结果通过方差分析发现不同方法对指纹图谱蛋白峰总量影响不显著,P=0.753> 0.05;但液氮甲酸方法获取的菌株指纹图谱平均特异峰数量最高为3.75个/株,图谱质量最好,更适用于大部分食用菌的前处理,同时发现云南省香格里拉县、四川省稻城县两地的松茸之间无法通过指纹图谱区分,表明此两地松茸含有相近的蛋白质组成。结论 MALDI-TOF-MS技术在大型食用菌甄别鉴定中具有开发运用前景。

Application of MALDI-TOF-MS Technology in Identification of Large-scale Edible Fungi

Objective To explore the application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry MALDI-TOF-MS in the identification of large-scale edible fungi by analyzing the protein profiles of common edible fungi such as Tricholoma matsutake(S. Ito & S. Imai) Singer, Agaricus blazei Murr, Tremella fuciformis Berk, Lentinus edodes(Berk.)Sing, Naematelia aurantialba(Bandoni & M. Zang) Millanes & Wedin, Dictyophora indusiata(Vent.ex Pers) Fisch and Armillaria mellea(Vahl) P. Kumm. Methods The edible fungus protein samples were extracted and obtained by four pretreatment methods: formic acid extraction method, liquid nitrogen formic acid method, ultrasonic formic acid crushing method, and liquid nitrogen formic acid ultrasonic crushing method, respectively. The fingerprint of samples was collected by MALDI-TOF-MS, and the fingerprint of different methods and samples were compared and cluster analyzed. Results Through analysis of variance, it was found that different methods had no significant effect on the total number of protein peaks in the fingerprint, P = 0.753 > 0.05;but the average number of specific peaks of the fingerprints of the strains obtained by the liquid nitrogen formic acid method was 3.75 per plant, and the quality of the map was the best It is more suitable for pretreatment of most edible fungi. At the same time, it is found that the Tricholoma matsutake(S. Ito & S. Imai) Singer from Shangri La County Yunnan Province and Daocheng County Sichuan Province cannot be distinguished from each other by fingerprints, indicating that the mushrooms in these two places contain similar protein composition. Conclusion MALDI-TOF-MS technology has a bright future in the identification of large-scale edible fungi.