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

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

Table II

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 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

Table III

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	##	++

1 Different small letters in the same column indicate significance at the 5% level

1 #– sparse colony density, ## – dense colony density, ### – densest colony density, + – weak mycelial growth, ++ – moderate mycelial growth, +++ – favorable mycelial growth, ++++ – strong mycelial growth

Table IV

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	##	+++

1 Different small letters in the same column indicate significance at the 5% level

1 # – sparse colony density, ## – dense colony density, ### – densest colony density, + – weak mycelial growth, ++ – moderate mycelial growth, +++ – favorable mycelial growth, ++++ – strong mycelial growth

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

Table V

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*	###	++++

1 Different small letters in the same column indicate significance at the 5% level

1 # – sparse colony density, ## – dense colony density, ### – densest colony density, + – weak mycelial growth, ++ – moderate mycelial growth, +++ – favorable mycelial growth, ++++ – strong mycelial growth

Table VI

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	#	+++

1 Small letter in the same column indicate significance at the 5% level

1 # – sparse colony density, ## – dense colony density, ### – densest colony density, + – weak mycelial growth, ++ – moderate mycelial growth, +++ – favorable mycelial growth, ++++ – strong mycelial growth

Table VII

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	##	+

1 Different small letters in the same column indicate significance at the 5% level

1 # – sparse colony density, ## – dense colony density, ### – densest colony density, + – weak mycelial growth, ++ – moderate mycelial growth, +++ – favorable mycelial growth, ++++ – strong mycelial growth, n.d. – no data for the experimental group

Table VIII

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	#	+++

1 Different small letters in the same column indicate significance at the 5% level

1 # – sparse colony density, ## – dense colony density, ### – densest colony density, + – weak mycelial growth, ++ – moderate mycelial growth, +++ – favorable mycelial growth, ++++ – strong mycelial growth

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

Table IX

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**	###	++

1 Different small letters in the same column indicate significance at the 5% level

1 # – sparse colony density, ## – dense colony density, ### – densest colony density, + – weak mycelial growth, ++ – moderate mycelial growth, +++ – favorable mycelial growth, ++++ – strong mycelial growth

Table X

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	###	+++

1 Different small letters in the same column indicate significance at the 5% level

1 # – sparse colony density, ## – dense colony density, ### – densest colony density, + – weak mycelial growth, ++ – moderate mycelial growth, +++ – favorable mycelial growth, ++++ – strong mycelial growth

Table XI

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

1 X₁ –X₈ code values are shown in Table I

Table XII

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

1 * – significant at 0.05 level, ** – significant at 0.01 level

Table XIII

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	##	+

1 # – sparse colony density, ## – dense colony density, ### – densest colony density, + – weak mycelial growth, ++ – moderate mycelial growth, +++ – favorable mycelial growth, ++++ – strong mycelial growth, n.d. – no data for the experimental group

Table XIV

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

1 A, B and C code values are shown in Table II

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

Table XV

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

1 * – significant at 0.05 level, ** – significant at 0.01 level

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

References

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Articles in this issue

DOI: https://doi.org/10.33073/pjm-2024-022 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331

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Language: English

Page range: 237 - 252

Submitted on: Feb 28, 2024

Accepted on: May 9, 2024

Published on: Jun 20, 2024

Published by: Polish Society of Microbiologists

In partnership with: Paradigm Publishing Services

Publication frequency: 4 issues per year

Keywords:

Phallus dongsun,

nutrient composition,

suspension agents,

temperature gradient,

mycelium growth rate

Related subjects:

Life sciences,

Microbiology and virology

© 2024 Chao Kang, Xuan Zheng, Wankun Wang, Weijun Zeng, Jing Wang, Zhongxuan Liu, Ling Yang, Fang Wang, Yan Zhu, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Volume 73 (2024): Issue 2 (June 2024)

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

Figures & Tables

Table I

Table II

Fig. 1.

Fig. 2.

Table III

Table IV

Fig. 3.

Fig. 4.

Table V

Table VI

Fig. 5.

Table VII

Fig. 6.

Table VIII

Fig. 7.

Fig. 8.

Table IX

Table X

Table XI

Table XII

Table XIII

Table XIV

Fig. 9.

Table XV

Fig. 10.

Fig. 11.

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