Biological Characteristics of Beauveria majiangensis Strain MJ1015 and Optimization of Solid Medium Technology for Sporulation Cover

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Biological Characteristics of Beauveria majiangensis Strain MJ1015 and Optimization of Solid Medium Technology for Sporulation

Polish Journal of Microbiology

Volume 72 (2023): Issue 4 (December 2023)

By: Xinghong Wang, Zhenxing Huang, Congchun Li, Man Liu, Guangming Yang, Lilin Luo, Yongchao Rao, Yan Shen and Jing Wang

Open Access

|Dec 2023

Figures & Tables

Effect of different culture conditions on colony growth rate and sporulation. Effect of different media (a–b), temperatures (c–d), and pH values (e–f) on colony growth rate and sporulation of Beauveria majiangensis strain MJ1015. Values shown are means ± the standard error. Different lowercase letters indicate significant differences between values (p < 0.05). PDA – potato dextrose agar medium, CZM – base medium, SDAY – Sabouraud dextrose agar with yeast extract medium, ¼ SDAY – ¼ Sabouraud dextrose agar with yeast extract medium.

a–f. Solid medium single-factor screening. Effect of different carbon sources (a–b), proportions of rice and wheat (c–d), nitrogen sources (e–f), and inorganic salts (g–h) on colony growth rate and sporulation of Beauveria majiangensis strain MJ1015. Values shown are means ± the standard error. Different lowercase letters indicate a significant difference between values (p < 0.05).

(a–d). Response surface diagrams showing the effects of rice and NaNO3 (contour (a) and 3D plots (b)), rice and K2HPO4 · 3H2O (contour (c) and 3D plots (d)), NaNO3 and K2HPO4 · 3H2O (contour (e) and 3D plots (f)) on the sporulation of Beauveria majiangensis strain MJ1015.

Plackett-Burman design and response values_

Order	Variable code								Y Sporulation (conidia/l) 1 × 10⁹
Order	X₁	X₂	X₃	X₄	X₅	X₆	X₇	X₈	Y Sporulation (conidia/l) 1 × 10⁹
1	1	1	−1	1	1	1	−1	−1	22.370 ± 1.833
2	−1	1	1	−1	1	1	1	−1	2.860 ± 0.282
3	1	−1	1	1	−1	1	1	1	4.267 ± 0.466
4	−1	1	−1	1	1	−1	1	1	0.923 ± 0.117
5	−1	−1	1	−1	1	1	−1	1	9.043 ± 1.283
6	−1	−1	−1	1	−1	1	1	−1	1.487 ± 0.289
7	1	−1	−1	−1	1	−1	1	1	33.453 ± 3.836
8	1	1	−1	−1	−1	1	−1	1	30.840 ± 2.155
9	1	1	1	−1	−1	−1	1	−1	11.113 ± 0.934
10	−1	1	1	1	−1	−1	−1	1	5.773 ± 0.636
11	1	−1	1	1	1	−1	−1	−1	6.950 ± 0.865
12	−1	−1	−1	−1	−1	−1	−1	−1	5.650 ± 1.211
13	0	0	0	0	0	0	0	0	10.628 ± 1.353
14	0	0	0	0	0	0	0	0	13.027 ± 1.477
15	0	0	0	0	0	0	0	0	19.129 ± 1.886

Results of regression analysis of Plackett-Burman design_

Source	Factor	df	Sporulation (Y)
Source	Factor	df	Sum of squares	Mean square	F-value	p-value
Model		5	11.69	2.34	7.84	0.0060**
X₁	0.7365	1	6.51	6.51	21.81	0.0016**
X₂	0.0313	1	0.0117	0.0117	0.0393	0.8477
X₄	-0.4789	1	2.75	2.75	9.22	0.0162*
X₅	0.0448	1	0.0241	0.0241	0.0807	0.7836
X₇	-0.4468	1	2.40	2.40	8.03	0.0220*
Residual		8	2.39	0.2985
Cor total		14	15.30

Range of different factors investigated with Plackett-Burman design_

Symbol	Variables	Experimental value
Symbol	Variables	Low (−1)	High (+1)
X₁	Rice (g/l)	27	33.75
X₂	Wheat (g/l)	13	16.25
X₃	Virtual 1	−1	1
X₄	NaNO₃ (g/l)	1	1.25
X₅	CaCO₃ (g/l)	1	1.25
X₆	Virtual 2	−1	1
X₇	K₂HPO₄ · 3H.O (g/l)	1	1 .25
X₈	Virtual 3	−1	1

Experimental design and results of steepest ascent path_

Level of factor	Factor coded value			Factor actual value			Sporulation (conidia/l) 1 × 10⁹
Level of factor	Rice	NaNO₃	K₂HPO₄ · 3H₂O	Rice	NaNO₃	K₂HPO₄ · 3H₂O	Sporulation (conidia/l) 1 × 10⁹
Step lengthΔ	1	0.125	0.125	3.375	0.125	0.125	n.d.
Origin	0	0.000	0.000	30.375	1.125	1.125	10.627 ± 0.342
Origin + 1Δ	1	0.125	0.125	33.750	1.044	1.049	9.043 ± 0.532
Origin + 2Δ	2	0.250	0.250	37.125	0.962	0.973	5.273 ± 0.136
Origin + 3Δ	3	0.375	0.375	40.500	0.881	0.898	4.837 ± 0.229
Origin + 4Δ	4	0.500	0.500	43.875	0.800	0.822	1.880 ± 0.218
Origin + 5Δ	5	0.625	0.625	47.250	0.719	0.746	0.730 ± 0.063
Origin + 6Δ	6	0.750	0.750	50.625	0.637	0.670	0.620 ± 0.008
Origin + 7Δ	7	0.875	0.875	54.000	0.556	0.594	29.487 ± 0.546
Origin + 8Δ	8	1.000	1.000	57.375	0.475	0.518	28.773 ± 0.540
Origin + 9Δ	9	1.125	1.125	60.750	0.393	0.443	35.627 ± 1.806
Origin + 10Δ	10	1.250	1.250	64.125	0.312	0.367	47.587 ± 4.056
Origin + 11Δ	11	1.375	1.375	67.500	0.231	0.291	9.917 ± 0.451
Origin + 12Δ	12	1.500	1.500	70.875	0.150	0.215	2.420 ± 0.079
Origin + 13Δ	13	1.625	1.625	74.250	0.068	0.139	3.967 ± 0.138

Response surface central composite design and corresponding results_

Run	Coded variable level			Y Sporulation (conidia/l) 1 × 10⁹
Run	A	B	C
1	−1	−1	−1	1.95 + 0.108
2	1	−1	−1	2.75 + 0.085
3	−1	1	−1	1.60 + 0.070
4	1	1	−1	2.34 + 0.098
5	−1	−1	1	1.75 + 0.060
6	1	−1	1	2.16 + 0.117
7	−1	1	1	1.45 + 0.062
8	1	1	1	1.55 + 0.076
9	−1.6818	0	0	2.13 + 0.061
10	1.6818	0	0	2.91 + 0.077
11	0	−1.6818	0	2.70 + 0.102
12	0	1.6818	0	1.23 + 0.038
13	0	0	−1.6818	2.58 + 0.076
14	0	0	1.6818	2.85 + 0.059
15	0	0	0	4.90 + 0.077
16	0	0	0	4.88 + 0.088
17	0	0	0	4.67 + 0.119
18	0	0	0	4.25 + 0.110
19	0	0	0	4.57 + 0.085
20	0	0	0	3.70 + 0.082

Response surface optimization results validation_

Medium	Colony growth (mm/d)	Sporulation (conidia/l) 1 × 10¹⁰
New medium	4.46 ± 0.04	4.54 ± 0.16
SDAY	2.41 ± 0.03	0.10 ± 0.02
PDA	2.28 ± 0.03	0.47 ± 0.04

ANOVA for response surface quadratic polynomial model_

Source	df	Sporulation (Y)
Source	df	Sum of squares	Mean square	F-value	p-value
Model	9	2.668 × 10²¹	2.965 × 10²⁰	15.93	< 0.0001	significant
A-Rice	1	8.275 × 10¹⁹	8.275 × 10¹⁹	4.450	0.0612
B-NaNO₃	1	1.256 × 10²⁰	1.256 × 10²⁰	6.750	0.0266
C-K₂HPO₄ · 3H₂O	1	1.180 × 10¹⁹	1.180 × 10¹⁹	0.6336	0.4445
AB	1	1.650 × 10¹⁸	1.650 × 10¹⁸	0.0886	0.7720
AC	1	1.343 × 10¹⁹	1.343 × 10¹⁹	0.7126	0.4155
BC	1	3.335 × 10¹⁷	3.335 × 10¹⁷	0.0179	0.8962
A²	1	8.529 × 10²⁰	8.529 × 10²⁰	45.810	< 0.0001
B²	1	1.339 × 10²¹	1.339 × 10²¹	71.900	< 0.0001
C²	1	7.069 × 10²⁰	7.069 × 10¹⁸	37.970	0.0001
Residual	10	1.862 × 10²⁰	1.862 × 10¹⁹
Lack of fit	5	8.170 × 10¹⁹	1.634 × 10¹⁹	0.7821	0.6030	not significant
Pure error	5	1.045 × 10²⁰	2.089 × 10¹⁹
Cor Total	19	2.855 × 10²¹
R²		0.9348
Adjusted R²		0.8761

Factors and levels of response surface central composite design_

Symbol	Variables	Code level
Symbol	Variables	−1.6828	−1	0	1	1.6818
A	Rice (g/l)	58.4489	60.75	64.125	67.50	69.8011
B	NaNO₃ (g/l)	0.1755	0.23	0.31	0.39	0.44459
C	K₂HPO₄ · 3H₂O (g/l)	0.2389	0.29	0.365	0.44	0.49119

DOI: https://doi.org/10.33073/pjm-2023-033 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331

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Language: English

Page range: 377 - 389

Submitted on: May 22, 2023

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Accepted on: Jul 24, 2023

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Published on: Dec 16, 2023

Published by: Polish Society of Microbiologists

In partnership with: Paradigm Publishing Services

Publication frequency: 4 issues per year

Keywords:

biocontrol agent,

biological characteristics,

spore sporulation

Related subjects:

Microbiology and virology

© 2023 Xinghong Wang, Zhenxing Huang, Congchun Li, Man Liu, Guangming Yang, Lilin Luo, Yongchao Rao, Yan Shen, Jing Wang, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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