长木蜂蜂粮防腐机理研究
摘要
本文以长木蜂Xylocopa tranquebarorum (Swederus)为研究对象,在对长木蜂筑巢和酿贮蜂粮行为研究的基础上,对长木蜂蜂粮酿贮过程中pH和花粉活力的变化、蜂粮粗提液的抑菌活性测定、蜂粮中10-羟基-2-癸烯酸(简称10-HDA)来源及其功能、蜂粮中功能微生物及其代谢产物等方面进行了系统研究,初步揭示了长木蜂蜂粮的防腐机理,并从长木蜂蜂粮中发现了具有高效抑菌活性的菌株,为进一步开发新的高活性天然防腐物质奠定了基础。主要结论如下:
长木蜂属于独栖性、多访花性昆虫,在南京地区1年发生1代,主要在竹子上筑巢,偶尔在芦苇上筑巢。酿贮蜂粮盛期在5月上旬,采集蜂粮最佳时期为5月上中旬。
长木蜂蜂粮主要由花粉和花蜜组成,在酿贮过程中添加雌蜂唾腺分泌物和体表微生物。雌蜂咽下腺分泌的10-HDA添加入蜂粮中,对花粉萌发有抑制作用。在长木蜂蜂粮中共分离微生物菌株68株,其中细菌43株,酵母17株,放线菌8株。在雌蜂体表分离到细菌、酵母菌和放线菌等微生物。蜂粮酿制过程中在细菌和酵母菌的作用下,蜂粮的pH值不断降低,从芍药花粉到3日龄蜂粮,pH值从6.19降至4.82,在此pH值下,很多细菌和霉菌均不能正常生长。
分离的68株菌株中有12株有较强抑菌活性,尤其是放线菌NF-34菌株对真菌和细菌均有抑菌活性;通过形态观察、生理生化特征以及16S rDNA序列分析方法,确定NF-34属于桑氏链霉菌Streptomyces sampsonii(Millaed and Burr)。采用活性追踪法对NF-34菌株的抑菌活性物质进行研究,明确该菌株代谢的抑菌物质主要是非水溶性Ⅰ型抗生素类。
In this paper, the anticorrosion mechanism of bee bread brewed by X.tranquebarorum was studied, the main research contents included nesting habit, the behavior of brewing bee bread, the changes of pH value and pollen activity during the process from corbicular pollen to bee bread, antibacterial activity of the raw extraction from bee bread, the source and function of 10-HDA in bee bread, function microorganisms and their metabolite extracted from bee bread and so on. The results layed a foundation for exploring new natural anticorrosion products. The main results were as follows.
X. tranquebarorum which belonged to solitary pollinates and many flowers visiting insect had 1 generation per year in Nanjing, it nested in bamboo usually, but it was found nesting in reed occasionally. Peak period of brewing bee bread was in early May, so the best period for collecting bee bread was in early or middle May.
The bee bread was composed of pollen and nectar mainly, which added by salivary gland secretion and microorganism from the body surface of female bee in the process of brewing. Because the 10-HDA from hypopharyngeal gland of female bees was added into bee bread, pollen germination was inhibited.
68 microorganism strains were separated from bee bread of X. tranquebarorum, including 43 bacterium strains, 17 yeasts strains, 8 actinomycete stains, the above-mentioned 3 types of microorganism were also separated from body surface of female bee. Because of the existence of bacteria and yeast, the pH value of bee bread decreased gradually in the process of brewing bee bread, from 6.19 (P. lactiflora pollen) to 4.82 (3-day bee bread). Under the condition of such pH value, many bacterium and moulds could not grow normally.
12 strains had stronger antibacterial activities among the above-mentioned 68 strains, actinomycete strain NF-34 had significant antibacterial activity to fungus and bacterium. According to its morphological and cultural characteristics, the physiological and biochemical properties and 16SrDNA sequence analysis, NF-34 strain was identified as Streptomyces sampsonii. The antibacterial metabolite of NF-34 strain had been identified asⅠAntibiotic of Water Insoluble by using Bioassay-guided Method.
长木蜂属于独栖性、多访花性昆虫,在南京地区1年发生1代,主要在竹子上筑巢,偶尔在芦苇上筑巢。酿贮蜂粮盛期在5月上旬,采集蜂粮最佳时期为5月上中旬。
长木蜂蜂粮主要由花粉和花蜜组成,在酿贮过程中添加雌蜂唾腺分泌物和体表微生物。雌蜂咽下腺分泌的10-HDA添加入蜂粮中,对花粉萌发有抑制作用。在长木蜂蜂粮中共分离微生物菌株68株,其中细菌43株,酵母17株,放线菌8株。在雌蜂体表分离到细菌、酵母菌和放线菌等微生物。蜂粮酿制过程中在细菌和酵母菌的作用下,蜂粮的pH值不断降低,从芍药花粉到3日龄蜂粮,pH值从6.19降至4.82,在此pH值下,很多细菌和霉菌均不能正常生长。
分离的68株菌株中有12株有较强抑菌活性,尤其是放线菌NF-34菌株对真菌和细菌均有抑菌活性;通过形态观察、生理生化特征以及16S rDNA序列分析方法,确定NF-34属于桑氏链霉菌Streptomyces sampsonii(Millaed and Burr)。采用活性追踪法对NF-34菌株的抑菌活性物质进行研究,明确该菌株代谢的抑菌物质主要是非水溶性Ⅰ型抗生素类。
In this paper, the anticorrosion mechanism of bee bread brewed by X.tranquebarorum was studied, the main research contents included nesting habit, the behavior of brewing bee bread, the changes of pH value and pollen activity during the process from corbicular pollen to bee bread, antibacterial activity of the raw extraction from bee bread, the source and function of 10-HDA in bee bread, function microorganisms and their metabolite extracted from bee bread and so on. The results layed a foundation for exploring new natural anticorrosion products. The main results were as follows.
X. tranquebarorum which belonged to solitary pollinates and many flowers visiting insect had 1 generation per year in Nanjing, it nested in bamboo usually, but it was found nesting in reed occasionally. Peak period of brewing bee bread was in early May, so the best period for collecting bee bread was in early or middle May.
The bee bread was composed of pollen and nectar mainly, which added by salivary gland secretion and microorganism from the body surface of female bee in the process of brewing. Because the 10-HDA from hypopharyngeal gland of female bees was added into bee bread, pollen germination was inhibited.
68 microorganism strains were separated from bee bread of X. tranquebarorum, including 43 bacterium strains, 17 yeasts strains, 8 actinomycete stains, the above-mentioned 3 types of microorganism were also separated from body surface of female bee. Because of the existence of bacteria and yeast, the pH value of bee bread decreased gradually in the process of brewing bee bread, from 6.19 (P. lactiflora pollen) to 4.82 (3-day bee bread). Under the condition of such pH value, many bacterium and moulds could not grow normally.
12 strains had stronger antibacterial activities among the above-mentioned 68 strains, actinomycete strain NF-34 had significant antibacterial activity to fungus and bacterium. According to its morphological and cultural characteristics, the physiological and biochemical properties and 16SrDNA sequence analysis, NF-34 strain was identified as Streptomyces sampsonii. The antibacterial metabolite of NF-34 strain had been identified asⅠAntibiotic of Water Insoluble by using Bioassay-guided Method.
引文
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