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Biological Characteristics of the Mycelium and Optimization of the Culture Medium for Phallus dongsun

Chao Kang
Chao Kang
, 
Xuan Zheng
Xuan Zheng
, 
Wankun Wang
Wankun Wang
, 
Weijun Zeng
Weijun Zeng
, 
Jing Wang
Jing Wang
, 
Zhongxuan Liu
Zhongxuan Liu
, 
Ling Yang
Ling Yang
, 
Fang Wang
Fang Wang
 et 
Yan Zhu
Yan Zhu
   | 20 juin 2024
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Article Category: Original Paper
Publié en ligne: 20 juin 2024
Pages: 237 - 252
Reçu: 28 févr. 2024
Accepté: 09 mai 2024
DOI: https://doi.org/10.33073/pjm-2024-022
Mots clés
© 2024 Chao Kang et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Effects of natural substances on the growth of Phallus dongsun. A – control group, B – peeled potato, C – carrot powder, D – wood powder, E – wheat bran, F – potato powder, G – corn starch, H – wheat starch
Effects of natural substances on the growth of Phallus dongsun. A – control group, B – peeled potato, C – carrot powder, D – wood powder, E – wheat bran, F – potato powder, G – corn starch, H – wheat starch

Fig. 2.

Effects of composite natural substances on the growth of Phallus dongsun. A – FA0, B – FA1, C – FA2, D – FA3, E – FA4, F – FA5, G – FA6, H – FA7
Effects of composite natural substances on the growth of Phallus dongsun. A – FA0, B – FA1, C – FA2, D – FA3, E – FA4, F – FA5, G – FA6, H – FA7

Fig. 3.

Effects of suspension agents on the physical state of the medium and the growth of Phallus dongsun. A – control group, B – sodium carboxylmethyl cellulose, C – sodium alginate, D – dimethyl sulfoxide, E – Tween 80, F – gum arabic, G – xanthan gum
Effects of suspension agents on the physical state of the medium and the growth of Phallus dongsun. A – control group, B – sodium carboxylmethyl cellulose, C – sodium alginate, D – dimethyl sulfoxide, E – Tween 80, F – gum arabic, G – xanthan gum

Fig. 4.

Effects of xanthan gum concentration on the physical state of the medium and the growth of Phallus dongsun. A – Control group, B – 0.5 g/l, C – 1.0 g/l, D – 1.5 g/l, E – 2.0 g/l, F – 2.5 g/l, G – 3.0 g/l
Effects of xanthan gum concentration on the physical state of the medium and the growth of Phallus dongsun. A – Control group, B – 0.5 g/l, C – 1.0 g/l, D – 1.5 g/l, E – 2.0 g/l, F – 2.5 g/l, G – 3.0 g/l

Fig. 5.

Effects of carbon sources on the growth of Phallus dongsun. A – control group, B – glucose, C – fructose, D – sucrose, E – maltose, F – lactose, G – brown sugar, H – cellulose, I – xylogen, J – chitosan
Effects of carbon sources on the growth of Phallus dongsun. A – control group, B – glucose, C – fructose, D – sucrose, E – maltose, F – lactose, G – brown sugar, H – cellulose, I – xylogen, J – chitosan

Fig. 6.

Effects of nitrogen sources on the growth of Phallus dongsun. A – control group, B – peptone, C – yeast powder, D – yeast extract, E – beef extract, F – soybean powder, G – KNO3, G – carbamide, I – NH4Cl, J – (NH4)2SO4, K – NaNO3
Effects of nitrogen sources on the growth of Phallus dongsun. A – control group, B – peptone, C – yeast powder, D – yeast extract, E – beef extract, F – soybean powder, G – KNO3, G – carbamide, I – NH4Cl, J – (NH4)2SO4, K – NaNO3

Fig. 7.

Effects of inorganic salts on the growth of Phallus dongsun. A – control group, B – KH2PO4, C – K2HPO4 · 3H2O, D – MgSO4 · 7H2O, E – MgCl2 · 6H2O, F – ZnSO4, G – CaCl2, H – CaSO4 · 2H2O, I – Fe2(SO4)3, J – CaCO3, K – NH4Cl, L – KNO3
Effects of inorganic salts on the growth of Phallus dongsun. A – control group, B – KH2PO4, C – K2HPO4 · 3H2O, D – MgSO4 · 7H2O, E – MgCl2 · 6H2O, F – ZnSO4, G – CaCl2, H – CaSO4 · 2H2O, I – Fe2(SO4)3, J – CaCO3, K – NH4Cl, L – KNO3

Fig. 8.

Effects of composite inorganic salts on the growth of Phallus dongsun. A – FM-A, B – FM-B, C – FM-C, D – FM-D, E – FM-E, F – FM-F, G – FM-G, H – FM-H
Effects of composite inorganic salts on the growth of Phallus dongsun. A – FM-A, B – FM-B, C – FM-C, D – FM-D, E – FM-E, F – FM-F, G – FM-G, H – FM-H

Fig. 9.

Two-dimensional contour plots of response surface on mycelial growth of Phallus dongsun. a – soybean powder vs. ZnSO4, b – soybean powder vs. NH4Cl, c – ZnSO4 vs. NH4Cl
Two-dimensional contour plots of response surface on mycelial growth of Phallus dongsun. a – soybean powder vs. ZnSO4, b – soybean powder vs. NH4Cl, c – ZnSO4 vs. NH4Cl

Fig. 10.

Effects of different temperature on colony diameter and mycelial growth rate of Phallus dongsun.
Effects of different temperature on colony diameter and mycelial growth rate of Phallus dongsun.

Fig. 11.

Effects of different temperature on mycelial growth of Phallus dongsun. A – 5°C, B – 10°C, C – 15°C, D – 20°C, E – 25°C, F – 30°C, G – 35°C
Effects of different temperature on mycelial growth of Phallus dongsun. A – 5°C, B – 10°C, C – 15°C, D – 20°C, E – 25°C, F – 30°C, G – 35°C

Effects of xanthan gum concentration on the physical state of the medium and the growth of Phallus dongsun.

Xanthan gum (g/l) Medium physical state Colony diameter (mm) Mycelium growth rate (mm/d) Colony density Hyphal growth
Control group Sediment 60.41 ± 7.95a 0.98 ± 0.14a ## +++
0.5 Stable suspension, Suitable viscosity 65.34 ± 6.52a 1.08 ± 0.12a ## +++
1.0 Stable suspension, Suitable viscosity 66.04 ± 5.46a 1.09 ± 0.10a ## +++
1.5 Stable suspension, Higher viscosity 62.63 ± 7.24a 1.03 ± 0.13a ## +++
2.0 Stable suspension, Higher viscosity 61.06 ± 5.46a 1.00 ± 0.10a ## +++
2.5 Stable suspension, high viscosity 56.23 ± 6.40a 0.91 ± 0.11a ## +++
3.0 Stable suspension, high viscosity 60.68 ± 4.23a 0.99 ± 0.08a # +++

Range of factors investigated using Plackett-Burman design.

Symbol Variables Experimental value
Low (-1) High (+1)
X1 Glucose (g/l) 10 15
X2 Wheat starch (g/l) 5 7.5
X3 Virtual 1 -1 +1
X4 Carrot powder (g/l) 5 7.5
X5 Soybean powder (g/l) 10 15
X6 Virtual 2 -1 +1
X7 ZnSO4 (g/l) 0.5 0.75
X8 NH4Cl (g/l) 0.5 0.75
X9 Virtual 3 -1 +1

Effects of composite inorganic salt on the growth of Phallus dongsun.

Test group Inorganic salt (g/l) Colony diameter (mm) Mycelium growth rate (mm/d) Colony density Hyphal growth
ZnSO4 Fe2(SO4)3 NH4Cl
FM-A 0 0 0 42.23 ± 8.15b 0.66 ± 0.15b ## ++
FM-B 1 0 0 50.04 ± 4.62ab 0.80 ± 0.08ab ### +++
FM-C 0 1 0 39.01 ± 2.45b 0.61 ± 0.04b ## ++
FM-D 0 0 1 39.07 ± 0.24ab 0.61 ± 0.00ab ## ++
FM-E 0.5 0.5 0 49.51 ± 6.03ab 0.79 ± 0.11ab ### ++
FM-F 0.5 0 0.5 61.10 ± 7.26a 1.00 ± 0.13a ### ++++
FM-G 0 0.5 0.5 50.05 ± 8.99ab 0.80 ± 0.16ab ### ++
FM-H 0.5 0.5 0.5 44.52 ± 5.92b 0.71 ± 0.11b ### +++

Effects of carbon sources on the growth of Phallus dongsun.

Carbon source (g/l) Colony diameter (mm) Mycelium growth rate (mm/d) Colony density Hyphal growth
Control group 36.84 ± 1.29bcd 0.56 ± 0.02bcd ## ++
Glucose 48.32 ± 3.63a 0.77 ± 0.06a ### +++
Fructose 37.92 ± 6.61bcd 0.58 ± 0.12bcd ## ++
Sucrose 32.43 ± 3.27d 0.49 ± 0.06d ## +++
Maltose 48.18 ± 2.97ab 0.77 ± 0.05ab ### +++
Lactose 37.13 ± 6.36bcd 0.57 ± 0.11bcd ### ++
Brown sugar 36.57 ± 4.02cd 0.56 ± 0.07cd ## +++
Cellulose 47.89 ± 5.17abc 0.76 ± 0.09abc ## +++
Xylogen 0.00 ± 0.00f 0.00 ± 0.00f n.d. n.d.
Chitosan 18.07 ± 6.54e 0.23 ± 0.12e ## +

ANOVA for response surface quadratic polynomial model.

Source df Sum of squares Means quares F-value p-value (Prob > F)
Modle 215.88 9 23.99 13.54 0.0002**
A – Soybean powder 143.14 1 143.14 80.81 < 0.0001**
B – ZnSO4 17.91 1 17.91 10.11 0.0098**
C – NH4Cl 0.6925 1 0.6925 0.3910 0.5458
AB 0.1922 1 0.1922 0.1085 0.7487
AC 10.44 1 10.44 5.89 0.0356*
BC 5.22 1 5.22 2.94 0.1169
A2 28.37 1 28.37 16.02 0.0025**
B2 1.46 1 1.46 0.8214 0.3861
C2 13.22 1 13.22 7.46 0.0211*
Residual 17.71 10 1.77
Lack of Fit 5.81 5 1.16 0.4882 0.7750
Pure Error 11.90 5 2.38
Cor Total 233.60 19
R2 0.9242
Adjusted R2 0.8559

Experimental design and results of the steepest ascent path.

Factor level Inorganic salt (g/l) Colony diameter (mm) Mycelium growth rate (mm/d) Colony density Hyphal growth
ZnSO4 NH4Cl
Step size (△) 0.125 0.073 n.d. n.d. n.d. n.d.
Origin (O) 0.5 0.5 48.13 ± 3.00 0.77 ± 0.05 ### +++
O + 1 △ 0.625 0.573 52.99 ± 5.85 0.85 ± 0.10 ### +++
O + 2 △ 0.750 0.646 54.20 ± 7.01 0.87 ± 0.13 ### +++
O + 3 △ 0.875 0.719 61.07 ± 2.20 1.00 ± 0.04 ### ++++
O + 4△ 1.000 0.792 60.43 ± 10.18 0.99 ± 0.18 ### ++++
O + 5△ 1.125 0.865 59.56 ± 4.15 0.88 ± 0.30 ### ++++
O + 6△ 1.250 0.938 48.82 ± 4.68 078 ± 0.08 ### ++++
O + 7△ 1.375 1.011 51.52 ± 9.49 0.83 ± 0.17 ### +++
O + 8 △ 1.500 1.084 32.24 ± 6.24 0.48 ± 0.11 ### ++
O + 9△ 1.625 1.157 35.39 ± 12.32 0.54 ± 0.22 ### +++
O + 10△ 1.75 1.230 29.47 ± 15.78 0.43 ± 0.28 ## +
O + 11△ 1.875 1.303 26.49 ± 8.02 0.38 ± 0.14 ## +
O + 12△ 2.00 1.376 23.38 ± 7.40 0.32 ± 0.13 ## +

Plackett-Burman design and response values.

Run Variables Y Colony diameter (mm)
X1 X2 X3 X4 X5 X6 X7 X8
1 1 1 -1 1 1 1 -1 -1 49.92 ± 3.47
2 -1 1 1 -1 1 1 1 -1 59.97 ± 6.54
3 1 -1 1 1 -1 1 1 1 74.45 ± 3.25
4 -1 1 -1 1 1 -1 1 1 60.35 ± 2.28
5 -1 -1 1 -1 1 1 -1 1 59.05 ± 4.68
6 -1 -1 -1 1 -1 1 1 -1 61.95 ± 5.93
7 1 -1 -1 -1 1 -1 1 1 63.52 ± 4.59
8 1 1 -1 -1 -1 1 -1 1 58.35 ± 4.82
9 1 1 1 -1 -1 -1 1 -1 62.39 ± 4.49
10 -1 1 1 1 -1 -1 -1 1 53.93 ± 6.15
11 1 -1 1 1 1 -1 -1 -1 50.62 ± 4.74
12 -1 -1 -1 -1 -1 -1 -1 -1 48.70 ± 2.69

Results of the regression analysis of the Plackett–Burman design.

Source Coefficient estimate df Sum of squares Means quares F–value p –value (Prob > F)
Model 58.60 6 486.42 81.07 5.14 0.0464*
X1 1.28 1 19.53 19.53 1.24 0.3162
X2 -1.11 1 14.90 14.90 0.9453 0.3756
X4 -0.0640 1 0.0491 0.0491 0.0031 0.9576
X5 -1.36 1 22.24 22.24 1.41 0.2882
X7 5.17 1 321.04 321.04 20.37 0.0063**
X8 3.01 1 108.66 108.66 6.90 0.0468*
Residual 5 78.79
Cor Total 11 565.20

Effects of nitrogen sources on the growth of Phallus dongsun.

Nitrogen source (g/l) Colony diameter (mm) Mycelium growth rate (mm/d) Colony density Hyphal growth
Control group 36.01 ± 5.30c 0.55 ± 0.09c ## +++
Peptone 19.16 ± 4.26cd 0.25 ± 0.08cd ## +
Yeast powder 9.16 ± 2.35cd 0.07 ± 0.04cd ## +
Yeast extract 5.72 ± 0.63d 0.01 ± 0.01d # +
Beef extract 39.86 ± 5.18* 0.62 ± 0.09* ## ++
Soybean powder 44.18 ± 5.17* 0.69 ± 0.09* ### +++
KNO3 40.62 ± 4.33* 0.63 ± 0.08* ## +++
Carbamide 5.70 ± 0.16d 0.01 ± 0.00d # +
NH4Cl 37.73 ± 5.98* 0.58 ± 0.11* ## ++
(NH4)2SO4 35.66 ± 3.32b 0.54 ± 0.06b ## ++
NaNO3 46.49 ± 5.31a 0.74 ± 0.09a # +++

Effects of suspension agents on the physical state of the medium and the growth of Phallus dongsun.

Suspension agent Medium physical state Colony diameter (mm) Mycelium growth rate (mm/d) Colony density Hyphal growth
Control group Sediment 44.45 ± 6.Γ’ 0.70 ± 0.11** ## +++
Sodium carboxylmethyl cellulose (g/l) Unstable suspension 45.05 ± 8.48* 0.72 ± 0.15* ## ++++
Sodium alginate (g/l) Unstable suspension 44.10 ± 8.64* 0.70 ± 0.15* ### ++++
Dimethyl sulfoxide (ml/l) Sediment 33.11 ± 8.35c 0.50 ± 0.15c ## ++++
Tween 80 (ml/l) Sediment 38.95 ± 6.89bc 0.61 ± 0.12bc # ++++
Gum arabic (g/l) Unstable suspension 50.73 ± 5.4a 0.82 ± 0.1a ## ++++
Xanthan gum (g/l) Stable suspension, high viscosity 48.03 ± 6.06* 0.77 ± 0.11* ### ++++

Effects of inorganic salts on the growth of Phallus dongsun.

Inorganic salt (g/l) Colony diameter (mm) Mycelium growth rate (mm/d) Colony density Hyphal growth
Control group 40.00 ± 7.53abc 0.62 ± 0.13abc ## +++
KH2PO4 33.66 ± 8.64bcd 0.51 ± 0.15bcd ## +++
K2HPO4·3H2O 36.14 ± 5.64bcd 0.55 ± 0.10bcd ### ++
MgSO4·7H2O 29.51 ± 4.16cd 0.43 ± 0.07cd ## ++
MgCl2·6H2O 39.37 ± 4.56abc 0.61 ± 0.08abc ## ++
ZnSO4 50.14 ± 9.16a 0.80 ± 0.16a ### ++++
CaCl2 37.11 ± 8.19abc 0.57 ± 0.15abc ### +++
CaSO4·2H2O 37.09 ± 4.72abcd 0.57 ± 0.08*cd # ++
Fe2(SO4)3 41.85 ± 9.97abc 0.65 ± 0.18abc ### ++++
CaCO3 20.79 ± 2.32d 0.28 ± 0.04d # +
NH4Cl 48.00 ± 7.14ab 0.76 ± 0.13* ### +++
KNO3 40.40 ± 6.27abc 0.63 ± 0.11** ### ++

Effects of natural substances on the growth of Phallum dongsun.

Natural substance Colony diameter (mm) Mycelium growth rate (mm/d) Colony density Hyphal growth
Control group 29.64 ± 7.07de 0.44 ± 0.13de # ++
Peeled potato 20.75 ± 0.79e 0.28 ± 0.01e ## +
Carrot powder 42.17 ± 3.23bcd 0.66 ± 0.06bcd ### +++
Wood powder 54.75 ± 5.1a 0.88 ± 0.09a ## +++
Wheat bran 52.71 ± 7.22ab 0.85 ± 0.13ab ## ++++
Potato starch 37.63 ± 3.63bcd 0.58 ± 0.06bcd # +
Corn starch 37.58 ± 5.35bcd 0.58 ± 0.10bcd # +
Wheat starch 48.12 ± 9.48abc 0.77 ± 0.17abc ## ++

Factors and levels of response surface central composite design.

Symbol Variables Code level
-1.6828 –1 0 1 1.6818
A Soybean powder (g/l) 5.80 7.50 10 12.50 14.20
B ZnSO4 (g/l) 0.45 0.63 0.88 1.13 1.30
C NH4Cl (g/l) 0.47 0.57 0.72 0.86 0.98

Effects of composite natural substances on the growth of Phallus dongsun.

Test group Natural substance (g/l) Colony diameter (mm) Mycelium growth rate (mm/d) Colony density Hyphal growth
Peeled potato Wheat starch Carrot powder Wood powder
FA0 200 0 0 0 26.78 ± 4.76c 0.38 ± 0.09c ### ++
FA1 0 20 0 0 56.19 ± 6.44b 0.91 ± 0.12b ## +++
FA2 0 0 20 0 51.57 ± 7.10b 0.83 ± 0.13b ## +++
FA3 0 0 0 20 34.34 ± 4.57c 0.52 ± 0.08c # ++
FA4 0 10 0 10 67.07 ± 4.79a 1.10 ± 0.09a # +++
FA5 0 10 10 0 52.35 ± 5.21b 0.84 ± 0.09b ### ++++
FA6 0 0 10 10 56.19 ± 8.14b 0.91 ± 0.15b # ++++
FA7 0 10 10 10 50.66 ± 9.46b 0.81 ± 0.17b ## +++

Response surface central composite design and matching results.

Run Variables Y Colony diameter (mm)
A B C
 1 -1 -1 -1 64.56 ± 5.88
 2 1 -1 -1 54.69 ± 4.98
 3 -1 1 -1 62.58 ± 7.79
 4 1 1 -1 54.35 ± 7.41
 5 -1 -1 1 63.78 ± 5.37
 6 1 -1 1 59.50 ± 4.29
 7 -1 1 1 59.59 ± 9.21
 8 1 1 1 54.91 ± 3.34
 9 -1.6818 0 0 63.33 ± 3.93
10 1.6818 0 0 53.13 ± 4.86
11 0 -1.6818 0 62.65 ± 4.13
12 0 1.6818 0 59.95 ± 4.14
13 0 0 -1.6818 60.88 ± 1.57
14 0 0 1.6818 58.10 ± 4.80
15 0 0 0 62.62 ± 2.36
16 0 0 0 61.43 ± 1.60
17 0 0 0 61.72 ± 7.81
18 0 0 0 64.65 ± 4.31
19 0 0 0 59.99 ± 2.18
20 0 0 0 61.76 ± 1.83
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