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MTT法检测滑菇培养基在不同酸碱度条件下菌丝生物量的研究 .doc
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MTT法检测滑菇培养基在不同酸碱度条件下菌丝生物量的研究 .doc
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摘 要
本次实验目的是利用 MTT 法来检测滑菇培养基菌丝与琥珀酸脱氢酶
(SDH)活性之间的关系,从而推出滑菇培养基在不同酸碱度时的菌丝生物量。
实验以滑菇 5188 菌种为实验材料,首先通过筛选找到合适的培养基配方和菌丝
培养方法,其次将液体培养好的滑菇菌丝球分别称取 0.05g、0.1g、0.15g、
0.2g、0.25g、0.3g、0.35g、0.4g 利用 MTT 法检测出琥珀酸脱氢酶(SDH)与滑
菇菌丝之间的标准曲线,以石灰(0%、1%、2%、3%、4%、5%、6%)来调节固体培
养基的酸碱度,然后以此曲线为标准得出滑菇固体培养基菌丝在不同酸碱条件下
其菌丝生物量是否有变化。实验结果表明:石灰量为 1%-3%的区间,菌丝生物量
总体呈上升趋势,在石灰量为 3%时达到最高。而当石灰量达到 4%及以上时,菌
丝生物量有所下降。所以,培养基石灰含量 3%最适合菌丝生长。本次实验是为 MTT
法测定食用菌菌丝生物量提供理论参考。
关键词:滑菇;MTT;菌丝生物量;琥珀酸脱氢酶
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Abstract
The purpose of this experiment is to use the MTT method to detect the
relationship between the mycelium of the mushroom medium and the activity of
succinate dehydrogenase (SDH), so as to introduce the mycelial biomass of the
mushroom medium at different pH levels. In the experiment, 5188 strains of sliding
mushrooms were used as experimental materials. First, the suitable medium
formulation and mycelium cultivation method were found through screening.
Secondly, the liquid cultured sliding mushroom mycelium balls were weighed 0.05g,
0.1g, 0.15g, 0.2 respectively. g, 0.25g, 0.3g, 0.35g, 0.4g using the MTT method to
detect the standard curve between succinate dehydrogenase (SDH) and the mushroom
hyphae, with lime (0%, 1%, 2%, 3 %, 4%, 5%, 6%) to adjust the pH of the solid
medium, and then use this curve as a standard to obtain whether the mycelial biomass
of the mycelia of the solid medium of the pleurotus ostreatus has changed under
different acid and alkali conditions. The experimental results show that: in the range
of 1% -3% of lime, the mycelial biomass generally shows an upward trend, reaching
the highest when the amount of lime is 3%. When the amount of lime reached 4% and
above, the mycelial biomass decreased. Therefore, the medium with a lime content of
3% is most suitable for mycelial growth. This experiment is to provide a theoretical
reference for the MTT method to determine the biomass of edible fungus mycelium.
Key words:Pholiota nameko;MTT;Mycelial biomass;Succinate dehydrogenase
![](https://csdnimg.cn/release/download_crawler_static/88343613/bg3.jpg)
目 录
1 引 言 ........................................................1
2 材料和方法 ......................................................2
2.1 材料............................................................2
2.1.1 供试菌种......................................................2
2.1.2 主要实验原料..................................................2
2.1.3 主要药品试剂..................................................3
2.1.4 仪器设备......................................................3
2.2 方法............................................................3
2.2.1 培养基配方及制作..............................................3
2.2.2 菌种的活化....................................................5
2.2.3 接种及培养....................................................5
2.2.4 MTT 的测定 ....................................................6
2.2.5 标准曲线的建立................................................7
2.2.6 固体培养基酸碱度测量..........................................7
2.2.7 菌丝日均生长速度测量..........................................7
2.2.8 液体菌丝浓度的计算............................................7
2.2.9 菌丝量的测定与计算............................................7
2.2.10 数据分析方法.................................................7
3 结果与分析 ......................................................8
3.1 菌丝生物量标准曲线的建立........................................8
3.2 固体培养基酸碱度对菌丝活性的影响...............................10
3.2.1 固体培养基酸碱度.............................................10
3.2.2 固体培养基总质量 .............................................11
3.2.3 固体培养基菌丝每天生长速度...................................12
3.2.4 固体培养基菌丝 SDH 活性 .......................................12
3.2.5 固体培养基每天菌丝生物量 .....................................13
4 结论与讨论 .....................................................14
参 考 文 献 ....................................................16
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