• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

大球盖菇液体菌种繁育工艺研究

曾志恒 戴建清 陈文智 曾辉 郭仲杰 蔡志欣

曾志恒,戴建清,陈文智,等. 大球盖菇液体菌种繁育工艺研究 [J]. 福建农业学报,2024,39(X):1−10
引用本文: 曾志恒,戴建清,陈文智,等. 大球盖菇液体菌种繁育工艺研究 [J]. 福建农业学报,2024,39(X):1−10
ZENG Z H, DAI J Q, CHEN W Z, et al. Study on liquid spawn reproductive technology of Stropharia rugosoannulata [J]. Fujian Journal of Agricultural Sciences,2024,39(X):1−10
Citation: ZENG Z H, DAI J Q, CHEN W Z, et al. Study on liquid spawn reproductive technology of Stropharia rugosoannulata [J]. Fujian Journal of Agricultural Sciences,2024,39(X):1−10

大球盖菇液体菌种繁育工艺研究

基金项目: 福建省属公益类科研院所基本科研专项(2021R1035002);宁夏回族自治区重点研发计划项目子课题(2022BBF02022-04);福建省农业科学院东西部协作项目(闽农科政[2023]8号)
详细信息
    作者简介:

    曾志恒(1984 —),男,硕士,助理研究员,主要从事食用菌工厂化制种技术研究,E-mail:67847563@qq.com

    通讯作者:

    蔡志欣(1983 —),男,硕士,副研究员,主要从事食用菌遗传育种及配套栽培技术研究,E-mail: 181351945@qq.com

  • 中图分类号: S646.1

Study on liquid spawn reproductive technology of Stropharia rugosoannulata

  • 摘要:   目的  本研究优化了大球盖菇液体菌种培养基,探究液体培养过程中生长规律,确立液体菌种繁育栽培种工艺参数。  方法  以大球盖菇8号为试验菌株,以菌丝体生物量为评价指标,采用单因素和正交试验L9(34)优化液体菌种培养基。通过测定液体菌种菌丝体生物量、还原糖和氨基氮含量、羧甲基纤维素酶、淀粉酶、酸性蛋白酶、漆酶胞外酶酶活生理生化指标,确立优化配方的液体菌种最优培养时间。以平均满袋时间为指标,确立液体菌种扩繁栽培种接种量,培养基配方颗粒度和碳氮比。  结果  优化得到大球盖菇液体菌种最优配方为葡萄糖20 g·L−1、小麦粉30 g·L−1、酵母粉0.75 g·L−1、磷酸二氢钾1.00 g·L−1、硫酸镁0.50 g·L−1、起始pH 5。培养第8 d时,大球盖菇菌丝体生物量最大,为1.66 g·hmL−1;液体培养过程中还原糖含量由12.23 mg·L−1降至1.38 mg·L−1,氨基氮含量由0.09 mg·mL−1降至0.06 mg·mL−1;羧甲基纤维素酶和淀粉酶酶活在第4 d最高,酶活分别为6.49 U和5.16 U,酸性蛋白酶酶活在第2 d 最高,酶活为1.80 U,漆酶酶活在第6 d 最大,酶活为1.63 U。液体菌种扩繁栽培种生产工艺参数:接种量为15 mL,菌包培养基配方颗粒度的粗细木屑比为7∶3,碳氮比为50∶1。  结论  大球盖菇液体菌种活性与上述指标具有一定的相关性,结合发酵液生理生化指标,判定第7d的液体菌种活力最高。利用大球盖菇液体菌种扩繁栽培种,平均满袋时间为23.7 d,缩短生长周期2.7 d。本研究建立配套的制种工艺,为大球盖菇栽培种工厂化生产技术奠定了基础。
  • 图  1  不同碳源对大球盖菇菌丝体生物量的影响

    不同小写字母表示差异显著水平(P<0.05),图2681012同。

    Figure  1.  Effects of different carbon sources onmycelial biomass of S.rugosoannulata

    The different lowercase letters indicate the significant level of difference at P<0.05. The same as Fig.2-6, 8, 10-12.

    图  2  不同氮源对大球盖菇菌丝体生物量的影响

    Figure  2.  Effects of different nitrogen sourceson mycelial biomass of S.rugosoannulata

    图  3  不同小麦粉添加量对大球盖菇菌丝体生物量的影响

    Figure  3.  Effects of adding amount of wheat flour on mycelial biomass of S.rugosoannulata

    图  4  不同酵母粉添加量对大球盖菇菌丝体生物量的影响

    Figure  4.  Effects of adding amount of yeast powder on mycelial biomass of S.rugosoannulata

    图  5  不同培养时间菌丝生物量的变化

    Figure  5.  Changes on mycelial biomass at different culture time

    图  6  不同时间培养液还原糖和氨基氮的变化

    Figure  6.  Changes on reducing sugar and amino nitrogen at different culture time

    图  4  不同起始pH对大球盖菇菌丝体生物量的影响

    Figure  4.  Effects of initial pH on mycelial biomass of S.rugosoannulata

    图  7  不同时间培养液胞外酶菌丝活性的变化

    Figure  7.  Changes on extracellular enzyme activities at different culture time

    图  8  不同菌种类型扩繁栽培种生长情况(左边:固体原种,右边:液体原种)

    Figure  8.  The growth of cultivation expanded by different spawn type (Left: solid spawn, right: liquid spawn)

    图  9  不同接种量对平均满袋时间的影响

    Figure  9.  Effect of different inoculation dose on the average hyphal pocketful time

    图  10  不同颗粒度对平均满袋时间的影响

    Figure  10.  Effect of different granularity on the average hyphal pocketful time

    图  11  不同碳氮比对平均满袋时间的影响

    Figure  11.  Effect of different carbon nitrogen ratio on the average hyphal pocketful time

    表  1  不同因素对菌丝体生物量影响的正交因素和水平

    Table  1.   Orthogonal factors and levels of influence of different factors on mycelial biomass

    水平
    Levels
    因素 Factors/(g·L−1)
    A葡萄糖
    A Gucose
    B小麦粉
    B Wheat flour
    C酵母粉
    C Yeast powder
    D 起始pH
    D Initial pH
    1 10 20 0.75 5
    2 15 25 1.00 6
    3 20 30 1.25 7
    下载: 导出CSV

    表  2  培养基优化的正交试验结果及极差分析

    Table  2.   Results of the orthogonal design experiments and range analysis for medium optimization

    试验号
    Run
    因素 Factors 菌丝体
    生物量
    Mycelial
    biomass/
    ( g/100 mL)
    A葡萄糖
    A Gucose
    B小麦粉
    B Wheat
    flour
    C酵母粉
    C Yeast
    powder
    D起始pH
    D Initial
    pH
    1 1 1 1 1 1.23
    2 1 2 3 3 1.12
    3 1 3 2 2 1.29
    4 2 1 3 2 1.18
    5 2 2 2 1 1.34
    6 2 3 1 3 1.44
    7 3 1 2 3 1.28
    8 3 2 1 2 1.56
    9 3 3 3 1 1.59
    K1j 3.64 3.68 4.23 4.17
    K2j 3.96 4.02 3.91 4.03
    K3j 4.43 4.33 3.89 3.84
    k1j 1.21 1.23 1.41 1.39
    k 2j 1.32 1.34 1.30 1.34
    k 3j 1.48 1.44 1.30 1.28
    极差 R 0.26 0.21 0.12 0.11
    优化水平
    Optimal level
    3 3 1 1
    下载: 导出CSV

    表  3  正交试验方差分析

    Table  3.   Variance analysis on the results of orthogonal experiment

    方差来源
    Source
    平方和
    Sum of
    squares
    自由度
    Degree of
    freedom
    均方
    Mean square
    F
    Fvalue
    显著性
    Significance
    A葡萄糖 0.41 2 0.20 80.13 **
    B小麦粉 0.28 2 0.14 54.28 **
    C酵母粉 0.06 2 0.03 11.01 **
    D起始pH 0.09 2 0.05 18.39 **
    误差Error 0.05 18 0.01
    总和Sum 49.92 27
    **表示差异显者(P<0.01)。
    **means significant differences (P<0.01).
    下载: 导出CSV

    表  4  不同碳氮比的菌包培养基配方

    Table  4.   Culture medium with different carbon nitrogen ratio in bag

    编号
    No
    杂木屑
    Wood
    chips/%
    麸皮
    Wheat
    bran/%
    花生粕
    Peanut
    meal/%
    碳酸钙
    calcium
    carbonate/%
    碳氮比
    Carbon
    nitrogen
    ratio
    1 83 14 2 1 70:1
    2 81 14 4 1 60:1
    3 80 14 5 1 50:1
    4 77 14 8 1 40:1
    5 73 14 12 1 30:1
    下载: 导出CSV
  • [1] 贺新生. 现代菌物分类系统[M]. 北京: 科学出版社, 2015: 238.
    [2] 黄年来. 大球盖菇的分类地位和特征特性 [J]. 食用菌, 1995, 17(6):11.

    HUANG N L. Taxonomic status and characteristics of Pleurotus ostreatus [J]. Edible Fungi, 1995, 17(6): 11. (in Chinese)
    [3] 姚春馨, 王小艳, 王小蓉, 等. 大球盖菇栽培研究现状与绿色发展前景 [J]. 中国食用菌, 2023, 42(6):90−97.

    YAO C X, WANG X Y, WANG X R, et al. Research progress on cultivation and green development of Stropharia rugosoannulata [J]. Edible Fungi of China, 2023, 42(6): 90−97. (in Chinese)
    [4] 边银丙. 大球盖菇栽培技术及其创新发展方向 [J]. 食药用菌, 2023, 31(06):370−377.

    BIAN Y B. Cultivation technology and its innovative development direction of Stropharia rugosoannulata [J]. Edible and Medicinal Mushrooms, 2023, 31(06): 370−377. (in Chinese)
    [5] 杨晓波, 蔡为明, 金群力, 等. 大球盖菇大棚设施栽培试验 [J]. 食药用菌, 2020, 28(5):328−331.

    YANG X B, CAI W M, JIN Q L, et al. Cultivation experiment of Stropharia rugoso-annulata in greenhouse [J]. Edible and Medicinal Mushrooms, 2020, 28(5): 328−331. (in Chinese)
    [6] 霍捷, 王卫平, 滑帆, 等. 大球盖菇栽培模式研究进展与发展方向探讨 [J]. 中国食用菌, 2020, 39(2):35−38.

    HUO J, WANG W P, HUA F, et al. Research progress and development direction of the cultivation model of Stropharia rugosoannulata [J]. Edible Fungi of China, 2020, 39(2): 35−38. (in Chinese)
    [7] LIU Y T, SUN J, LUO Z Y, et al. Chemical composition of five wild edible mushrooms collected from Southwest China and their antihyperglycemic and antioxidant activity [J]. Food and Chemical Toxicology: an International Journal Published for the British Industrial Biological Research Association, 2012, 50(5): 1238−1244. doi: 10.1016/j.fct.2012.01.023
    [8] LI X P, CUI W J, CUI Y F, et al. Stropharia rugoso-annulata acetylated polysaccharides alleviate NAFLD via Nrf2/JNK1/AMPK signaling pathways [J]. International Journal of Biological Macromolecules, 2022, 215: 560−570. doi: 10.1016/j.ijbiomac.2022.06.156
    [9] JIN M Z, ZHANG W Q, ZHANG X M, et al. Characterization, chemical modification and bioactivities of a polysaccharide from Stropharia rugosoannulata [J]. Process Biochemistry, 2023, 128: 30−39. doi: 10.1016/j.procbio.2023.02.008
    [10] WU J, SUZUKI T, CHOI J H, et al. An unusual sterol from the mushroom Stropharia rugosoannulata [J]. Tetrahedron Letters, 2013, 54(36): 4900−4902. doi: 10.1016/j.tetlet.2013.06.142
    [11] 李正鹏, 吴萍, 陆晓民. 大球盖菇液体菌种培养条件的研究 [J]. 中国林副特产, 2006, (4):12−14. doi: 10.3969/j.issn.1001-6902.2006.04.005

    LI Z P, WU P, LU X M. Study on the training condition of liquid culture Stropharia rugoso-annulata [J]. Forest by-Product and Speciality in China, 2006(4): 12−14. (in Chinese) doi: 10.3969/j.issn.1001-6902.2006.04.005
    [12] 曾健勇, 张国财, 赵博, 等. 磁化水培养对大球盖菇菌丝生长特性的影响 [J]. 食品工业科技, 2017, 38(20):110−114.

    ZENG J Y, ZHANG G C, ZHAO B, et al. Effect of magnetized water liquid cultivation on growth characteristics of Stropharia rugoso-annulata mycelium [J]. Science and Technology of Food Industry, 2017, 38(20): 110−114. (in Chinese)
    [13] 付婧璇, 耿丹萌, 高华, 等. 香蒲资源化开发中大球盖菇液体菌种的制备与应用 [J]. 河北大学学报(自然科学版), 2019, 39(1):76−79.

    FU J X, GENG D M, GAO H, et al. Preparation and application of liquid strains of Stropharia rugoso-annulata in the development of Typha resources [J]. Journal of Hebei University (Natural Science Edition), 2019, 39(1): 76−79. (in Chinese)
    [14] 刘克芳, 于国荣, 王华丽, 等. 大球盖菇液体菌种培养基的优化 [J]. 中国食用菌, 2021, 40(8):24−28.

    LIU K F, YU G R, WANG H L, et al. Optimization of liquid spawn formula of Stropharia rugosoannulata [J]. Edible Fungi of China, 2021, 40(8): 24−28. (in Chinese)
    [15] 杜瑞垚. 大球盖菇菌丝液体培养及液体菌种固化技术研究[D]. 武汉: 华中农业大学, 2023.

    DU R Y. Study on the Technology of Mycelia Liquid Culture and Liquid Spawn Solidification in Stropharia Rugosoannulata[D]. Wuhan: Huazhong Agricultural University, 2023. (in Chinese)
    [16] 曾志恒, 曾辉, 程翊, 等. 双孢蘑菇发酵液还原糖和总糖的含量测定 [J]. 中国食用菌, 2018, 37(6):40−43,49.

    ZENG Z H, ZENG H, CHENG Y, et al. Determination of reducing sugar and total sugar content in fermentation liquid of Agaricus bisporus [J]. Edible Fungi of China, 2018, 37(6): 40−43,49. (in Chinese)
    [17] 曾志恒, 曾辉, 程翊, 等. 双孢蘑菇发酵液氨基氮含量测定方法的研究 [J]. 食用菌, 2019, 41(5):77−80. doi: 10.3969/j.issn.1000-8357.2019.05.026

    ZENG Z H, ZENG H, CHENG Y, et al. Study on determination method of amino nitrogen content in fermentation broth of Agaricus bisporus [J]. Edible Fungi, 2019, 41(5): 77−80. (in Chinese) doi: 10.3969/j.issn.1000-8357.2019.05.026
    [18] 戴建清. 双孢蘑菇W192液体菌种摇瓶培养过程中的生理生化分析 [J]. 福建农业学报, 2021, 36(2):182−187.

    DAI J Q. Physiological and biochemical properties of Agaricus bisporus in shaking flask culture [J]. Fujian Journal of Agricultural Sciences, 2021, 36(2): 182−187. (in Chinese)
    [19] 田华. 现代菌物分类系统[M]. 北京: 化学工业出版社, 2019.
    [20] 刘金力, 唐琳, 陈思等. 大球盖菇液体菌种培养基碳氮源及培养条件的优化[J]. 黑龙江农业科学, 2019, (04): 94-99. [20] 刘金力, 唐琳, 陈思, 等. 大球盖菇液体菌种培养基碳氮源及培养条件的优化[J]. 黑龙江农业科学, 2019(4): 94-99.

    LIU J L, TANG L, CHEN S, et al. Optimization of carbon and nitrogen sources and culture conditions for liquid culture medium of Stropharia rugosoannulata [J]. Heilongjiang Agricultural Sciences, 2019, (04): 94-99. (in Chinese) LIU J L, TANG L, CHEN S, et al. Optimization of carbon and nitrogen sources and culture conditions of liquid culture medium of Stropharia rugosoannulata[J]. Heilongjiang Agricultural Sciences, 2019(4): 94-99. (in Chinese)
    [21] 周韬. 香菇液体菌种培养基优化及其菌种质量评价体系的建立[D]. 武汉: 华中农业大学, 2017.

    ZHOU T. Research on medium optimization and quality evaluation system of Lentinula edodes liquid spawn[D]. Wuhan: Huazhong Agricultural University, 2017. (in Chinese)
    [22] 曾志恒, 程翊, 曾辉, 等. 双孢蘑菇W192菌株液体发酵培养的研究 [J]. 福建农业学报, 2013, 28(8):763−769. doi: 10.3969/j.issn.1008-0384.2013.08.008

    ZENG Z H, CHENG Y, ZENG H, et al. Study on fermentation of Agaricus bisporus strain W192 [J]. Fujian Journal of Agricultural Sciences, 2013, 28(8): 763−769. (in Chinese) doi: 10.3969/j.issn.1008-0384.2013.08.008
    [23] 孙萌. 大球盖菇菌丝培养及胞外酶活性变化规律研究[D]. 延吉: 延边大学, 2013.

    SUN M. Mycelium culture and the variation regularity of extracellular enzyme activity of stropharia rugosoannulata Farlow[D]. Yanji: Yanbian University, 2013. (in Chinese)
    [24] 戴建成. 大球盖菇培养基质的优化及工厂化生产的初步探究[D]. 南京: 南京农业大学, 2021.

    DAI J C. Optimization of Medium Quality and Preliminary Study on Industrial Production of Stropharia Rugosoannulata[D]. Nanjing: Nanjing Agricultural University, 2021. (in Chinese)
    [25] 王升厚, 李玉双. 大球盖菇液体母种制备培养基碳氮源的优化 [J]. 北方园艺, 2008, (5):219−221.

    WANG S H, LI Y S. The optimization of carbonaceous compounds and nitrogenous substances for liquid culture media of Stropharia rugoso annulata [J]. Northern Horticulture, 2008(5): 219−221. (in Chinese)
    [26] 黄桂英, 杨林, 左天兴. 提高白僵菌培养基透气性的试验[J]. 云南林业科技, 1996, 25(2): 71-73. [26] 黄桂英, 杨林, 左天兴. 提高白僵菌培养基透气性的试验[J]. 云南林业科技, 1996(2): 71-73.

    HUANG G Y, YANG L, ZUO T X. A Test on Raising BreathbiIity to Beauveria bassiana medium[J]. Journal of West China Forestry Science, 1996, 25(2): 71-73. (in Chinese) HUANG G Y, YANG L, ZUO T X. A Test on Raising BreathbiIity to Beauveria bassiana medium[J]. Yunnan Forestry Science and Technology, 1996(2): 71-73. (in Chinese)
    [27] 曾志恒, 曾辉, 蔡志欣, 等. 双孢蘑菇新型颗粒原种配方的优化 [J]. 生物技术通报, 2021, 37(11):134−141.

    ZENG Z H, ZENG H, CAI Z X, et al. Formula optimization of the novel particle pre-culture spawn of Agaricus bisporus [J]. Biotechnology Bulletin, 2021, 37(11): 134−141. (in Chinese)
  • 加载中
图(12) / 表(4)
计量
  • 文章访问数:  25
  • HTML全文浏览量:  19
  • PDF下载量:  0
  • 被引次数: 0
出版历程
  • 收稿日期:  2024-04-12
  • 修回日期:  2024-06-14
  • 网络出版日期:  2024-07-10

目录

    /

    返回文章
    返回