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

Kang, Chao; Zheng, Xuan; Wang, Wankun; Zeng, Weijun; Wang, Jing; Liu, Zhongxuan; Yang, Ling; Wang, Fang; Zhu, Yan

doi:10.33073/pjm-2024-022

Figures & Tables

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

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

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

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

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

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

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

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

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

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

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 ± 7.95^a	0.98 ± 0.14^a	##	+++
0.5	Stable suspension, Suitable viscosity	65.34 ± 6.52^a	1.08 ± 0.12^a	##	+++
1.0	Stable suspension, Suitable viscosity	66.04 ± 5.46^a	1.09 ± 0.10^a	##	+++
1.5	Stable suspension, Higher viscosity	62.63 ± 7.24^a	1.03 ± 0.13^a	##	+++
2.0	Stable suspension, Higher viscosity	61.06 ± 5.46^a	1.00 ± 0.10^a	##	+++
2.5	Stable suspension, high viscosity	56.23 ± 6.40^a	0.91 ± 0.11^a	##	+++
3.0	Stable suspension, high viscosity	60.68 ± 4.23^a	0.99 ± 0.08^a	#	+++

Range of factors investigated using Plackett-Burman design_

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

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 ± 1.29^bcd	0.56 ± 0.02^bcd	##	++
Glucose	48.32 ± 3.63^a	0.77 ± 0.06^a	###	+++
Fructose	37.92 ± 6.61^bcd	0.58 ± 0.12^bcd	##	++
Sucrose	32.43 ± 3.27^d	0.49 ± 0.06^d	##	+++
Maltose	48.18 ± 2.97^ab	0.77 ± 0.05^ab	###	+++
Lactose	37.13 ± 6.36^bcd	0.57 ± 0.11^bcd	###	++
Brown sugar	36.57 ± 4.02^cd	0.56 ± 0.07^cd	##	+++
Cellulose	47.89 ± 5.17^abc	0.76 ± 0.09^abc	##	+++
Xylogen	0.00 ± 0.00^f	0.00 ± 0.00^f	n.d.	n.d.
Chitosan	18.07 ± 6.54^e	0.23 ± 0.12^e	##	+

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
Factor level	ZnSO₄	NH₄Cl	Colony diameter (mm)	Mycelium growth rate (mm/d)	Colony density	Hyphal growth
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)
Run	X₁	X₂	X₃	X₄	X₅	X₆	X₇	X₈	Y Colony diameter (mm)
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 ± 5.30^c	0.55 ± 0.09^c	##	+++
Peptone	19.16 ± 4.26^cd	0.25 ± 0.08^cd	##	+
Yeast powder	9.16 ± 2.35^cd	0.07 ± 0.04^cd	##	+
Yeast extract	5.72 ± 0.63^d	0.01 ± 0.01^d	#	+
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 ± 0.16^d	0.01 ± 0.00^d	#	+
NH₄Cl	37.73 ± 5.98*	0.58 ± 0.11*	##	++
(NH₄)₂SO₄	35.66 ± 3.32^b	0.54 ± 0.06^b	##	++
NaNO₃	46.49 ± 5.31^a	0.74 ± 0.09^a	#	+++

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 ± 8.35^c	0.50 ± 0.15^c	##	++++
Tween 80 (ml/l)	Sediment	38.95 ± 6.89^bc	0.61 ± 0.12^bc	#	++++
Gum arabic (g/l)	Unstable suspension	50.73 ± 5.4^a	0.82 ± 0.1^a	##	++++
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 ± 7.53^abc	0.62 ± 0.13abc	##	+++
KH₂PO₄	33.66 ± 8.64^bcd	0.51 ± 0.15^bcd	##	+++
K₂HPO₄·3H₂O	36.14 ± 5.64^bcd	0.55 ± 0.10^bcd	###	++
MgSO₄·7H₂O	29.51 ± 4.16^cd	0.43 ± 0.07^cd	##	++
MgCl₂·6H₂O	39.37 ± 4.56^abc	0.61 ± 0.08abc	##	++
ZnSO₄	50.14 ± 9.16^a	0.80 ± 0.16^a	###	++++
CaCl₂	37.11 ± 8.19^abc	0.57 ± 0.15abc	###	+++
CaSO₄·2H₂O	37.09 ± 4.72^abcd	0.57 ± 0.08*^cd	#	++
Fe₂(SO₄)₃	41.85 ± 9.97^abc	0.65 ± 0.18abc	###	++++
CaCO₃	20.79 ± 2.32^d	0.28 ± 0.04^d	#	+
NH₄Cl	48.00 ± 7.14^ab	0.76 ± 0.13*	###	+++
KNO₃	40.40 ± 6.27^abc	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 ± 7.07^de	0.44 ± 0.13^de	#	++
Peeled potato	20.75 ± 0.79^e	0.28 ± 0.01^e	##	+
Carrot powder	42.17 ± 3.23^bcd	0.66 ± 0.06^bcd	###	+++
Wood powder	54.75 ± 5.1^a	0.88 ± 0.09^a	##	+++
Wheat bran	52.71 ± 7.22^ab	0.85 ± 0.13ab	##	++++
Potato starch	37.63 ± 3.63^bcd	0.58 ± 0.06^bcd	#	+
Corn starch	37.58 ± 5.35^bcd	0.58 ± 0.10^bcd	#	+
Wheat starch	48.12 ± 9.48^abc	0.77 ± 0.17abc	##	++

Factors and levels of response surface central composite design_

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

Response surface central composite design and matching results_

Run	Variables			Y Colony diameter (mm)
Run	A	B	C	Y Colony diameter (mm)
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

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

Figures & Tables

Fig. 1.

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Fig. 6.

Fig. 7.

Fig. 8.

Fig. 9.

Fig. 10.

Fig. 11.

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

Range of factors investigated using Plackett-Burman design_

Effects of composite inorganic salt on the growth of Phallus dongsun_

Effects of carbon sources on the growth of Phallus dongsun_

ANOVA for response surface quadratic polynomial model_

Experimental design and results of the steepest ascent path_

Plackett-Burman design and response values_

Results of the regression analysis of the Plackett–Burman design_

Effects of nitrogen sources on the growth of Phallus dongsun_

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

Effects of inorganic salts on the growth of Phallus dongsun_

Effects of natural substances on the growth of Phallum dongsun_

Factors and levels of response surface central composite design_

Effects of composite natural substances on the growth of Phallus dongsun_

Response surface central composite design and matching results_

Paradigm

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