Due to a mass of hyperbranched β-glucan, glycoprotein and other active substances in sclerotia of Pleurotus tuber-regium, it has several biological functions, such as anti-tumor, immuno-modulatory, and so on. However, they could only be produced by bag cultivation, which has the disadvantages of a long cultivation period, low efficiency, and high cost. This work used a petri dish containing potato dextrose agar to cultivate P. tuber-regium sclerotia in a simple and fast method. The sclerotia could be optimally developed after 30 days cultivation at 25 ℃ and pH 5. Although the quantity of H2O2 in sclerotia cells was lower than that in mycelia, catalase activity had the opposite effect. The findings showed that optimum quantity of H2O2 in culture medium might cause intracellular oxidative stress and promote sclerotia development, whereas ascorbic acid could prevent sclerotia formation by removing intracellular oxidative stress. RT-qPCR experiment revealed that catalase activity, cell wall synthesis and the MAPK pathway were all involved in the production of sclerotia. The concentration of reactive oxygen species (ROS) in mycelial cells would increase when the external temperature, pH and redox environment changed. When the quantity of ROS reached a particular threshold, intracellular oxidative stress would ensue, triggering the cell to differentiate into sclerotium.
ZHOU Jun
,
ZHANG Zhuomin
,
WU Siyun
,
CHEN Lei
,
DING Zhongyang
. Cultivation of Pleurotus tuber-regium sclerotia via plate and its formation mechanism[J]. Food and Fermentation Industries, 2022
, 48(24)
: 84
-90
.
DOI: 10.13995/j.cnki.11-1802/ts.031309
[1] 黄飞龙. 虎奶菇多糖提取和抗胃癌活性研究及产品开发[D].南昌:南昌大学, 2018.
HUANG F L.The extraction and anti-gastric cancer activity of polysaccharide from Pleurotus tuber regium and product development[D].Nanchang:Nanchang University, 2018.
[2] CHEN L, XU W W, LIN S L, et al.Cell wall structure of mushroom sclerotium (Pleurotus tuber regium):Part 1.Fractionation and characterization of soluble cell wall polysaccharides[J].Food Hydrocolloids, 2014,36:189-195.
[3] CHEN L, CHEUNG P C K.Mushroom dietary fiber from the fruiting body of Pleurotus tuber-regium:Fractionation and structural elucidation of nondigestible cell wall components[J].Journal of Agricultural and Food Chemistry, 2014,62(13):2 891-2 899.
[4] 葛梦蝶, 代安然, 杨崇婧, 等.虎奶菇菌丝体细胞壁成分分析及多糖结构表征[J].食品与发酵工业, 2020,46(6):41-48.
GE M D, DAI A R, YANG C J, et al.Analysis on cell wall components of Pleurotus tuber regium mycelium and characterization of its polysaccharide structure[J].Food and Fermentation Industries, 2020,46(6):41-48.
[5] CHEN L, ZHANG B B, CHEN J L, et al.Cell wall structure of mushroom sclerotium (Pleurotus tuber-regium):Part 2.Fine structure of a novel alkali-soluble hyper-branched cell wall polysaccharide[J].Food Hydrocolloids, 2014,38:48-55.
[6] WONG S M, WONG K K, CHIU L C M, et al.Non-starch polysaccharides from different developmental stages of Pleurotus tuber-regium inhibited the growth of human acute promyelocytic leukemia HL-60 cells by cell-cycle arrest and/or apoptotic induction[J].Carbohydrate Polymers, 2007,68(2):206-217.
[7] CHEUNG P C, LEE M Y.Fractionation and characterization of mushroom dietary fiber (nonstarch polysaccharides) as potential nutraceuticals from sclerotia of Pleurotus tuber-regium (Fries) singer[J].Journal of Agricultural and Food Chemistry, 2000,48(8):3 148-3 151.
[8] ZHANG M, CHEUNG P C K, OOI V E C, et al.Evaluation of sulfated fungal β-glucans from the sclerotium of Pleurotus tuber-regium as a potential water-soluble anti-viral agent[J].Carbohydrate Research, 2004,339(13):2 297-2 301.
[9] SUN X Y, LIU D M, WANG Y Y, et al.Biogenesis of macrofungal Sclerotia:Influencing factors and molecular mechanisms[J].Applied Microbiology and Biotechnology, 2020,104(10):4 227-4 234.
[10] SMITH M E, HENKEL T W, ROLLINS J A.How many fungi make sclerotia?[J].Fungal Ecology, 2015,13:211-220.
[11] BAMIGBOYE C O, OLOKE J K, DAMES J F.Development of high yielding strain of Pleurotus tuber-regium:Fructification, nutritional and phylogenetic studies[J].Journal of Food Science and Technology, 2019,56(8):3 597-3 608.
[12] ROGER B, LUYEYE FELICIEN L, NGIALA GÉDÉON B, et al.Cultivation trial of an edible and medicinal mushroom species, Pleurotus tuber-regium (Rumph.ex Fr.) Singer 1951 (strain 190212) on various lignocellulosic substrates[J].Open Journal of Agricultural Research, 2021,1(2):84-93.
[13] ROLLINS J A, DICKMAN M B.pH signaling in Sclerotinia sclerotiorum:Identification of a pacC/RIM1 homolog[J].Applied and Environmental Microbiology, 2001,67(1):75-81.
[14] LIU H H, HUANG C C, LIN Y H, et al.Superoxide initiates the hyphal differentiation to microsclerotia formation of Macrophomina phaseolina[J].Microbiology Spectrum, 2022,10(1):e208421.
[15] PAPAPOSTOLOU I, SIDERI M, GEORGIOU C D.Cell proliferating and differentiating role of H2O2 in Sclerotium rolfsii and Sclerotinia sclerotiorum[J].Microbiological Research, 2014, 169(7-8):527-532.
[16] 汪浩德. 立枯丝核菌菌核形成过程中H2O2的作用及CAT的功能分析[D].沈阳:沈阳农业大学, 2018.
WANG H D.Role of H2O2 and function analysis of CAT in the sclerotia formation of Rhizoctonia solani[D].Shenyang:Shenyang Agricultural University, 2018.
[17] GEORGIOU C D, ZERVOUDAKIS G, PETROPOULOU K P.Ascorbic acid might play a role in the sclerotial differentiation of Sclerotium rolfsii[J].Mycologia, 2003,2(95):308-316.
[18] XING Y M C J S C.Nox gene expression and cytochemical localization of hydrogen peroxide in Polyporus umbellatus sclerotial formation[J].International Journal of Molecular Sciences, 2013,11(14):22 967-22 981.
[19] LI B, TIAN X, WANG C, et al.Swath label-free proteomics analyses revealed the roles of oxidative stress and antioxidant defensing system in sclerotia formation of Polyporus umbellatus[J].Scientific Reports, 2017,7:41283.
[20] CHEN C, HAREL A, GOROVOITS R, et al.MAPK regulation of sclerotial development in Sclerotinia sclerotiorum is linked with pH and cAMP sensing[J].Molecular Plant-Microbe Interactions, 2004,4(17):404-413.
[21] LI B, SUN L, XING Y, et al.Cell wall strengthening and remodeling provide new insights on sclerotia development of medicinal fungus Polyporus umbellatus[J].Research Square, 2020.DOI:10.21203/rs.3.rs-122799/v1.
