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

Xinghong Wang
Xinghong Wang
, 
Zhenxing Huang
Zhenxing Huang
, 
Congchun Li
Congchun Li
, 
Man Liu
Man Liu
, 
Guangming Yang
Guangming Yang
, 
Lilin Luo
Lilin Luo
, 
Yongchao Rao
Yongchao Rao
, 
Yan Shen
Yan Shen
 y 
Jing Wang
Jing Wang
   | 16 dic 2023
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Article Category: ORIGINAL PAPER
Publicado en línea: 16 dic 2023
Páginas: 377 - 389
Recibido: 22 may 2023
Aceptado: 24 jul 2023
DOI: https://doi.org/10.33073/pjm-2023-033
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© 2023 Xinghong Wang et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1

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.
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.

Fig. 2

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–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).

Fig. 3

(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.
(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
X1 X2 X3 X4 X5 X6 X7 X8
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)
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
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
Rice NaNO3 K2HPO4 · 3H2O Rice NaNO3 K2HPO4 · 3H2O
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
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)  
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
−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
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