Scientia Agricultura Sinica ›› 2020, Vol. 53 ›› Issue (14): 2974-2982.doi: 10.3864/j.issn.0578-1752.2020.14.020

• RESEARCH NOTES • Previous Articles    

Species Identification and Virulence Determination of Beauveria bassiana Strain BEdy1 from Ergania doriae yunnanus

ZHANG Lei(),JIA Qi,WU Wei,ZHAO LuPing,XUE Bing,LIU HuanHuan,SHANG Jing(),YONG TaiWen,LI Qing,YANG WenYu   

  1. College of Agronomy, Sichuan Agricultural University/Sichuan Key Laboratory for Major Crop Diseases/Sichuan Engineering Research Center for Crop Strip Intercropping System, Chengdu 611130
  • Received:2019-11-20 Accepted:2019-12-25 Online:2020-07-16 Published:2020-08-10
  • Contact: Jing SHANG E-mail:zhanglei5281@126.com;shangjing_edu@163.com

Abstract:

【Objective】Ergania doriae yunnanus has become one of the main pests in soybean fields in southern China. The objective of this study is to investigate the species and biological characteristics of an entomopathogenic fungus that naturally caused the death of E. d. yunnanus, determine the virulence of this fungal strain to E. d. yunnanus adults, and to provides a new way for biological control of E. d. yunnanus.【Method】The fungus from E. d. yunnanus was isolated and purified, and the fungal DNA was extracted for amplification of rDNA-ITS. Subsequently, BLAST alignment and phylogenetic tree construction were used to identify fungal species that caused the natural death of E. d. yunnanus. The growth rate and sporulation of the fungus were measured by cross method and hemocytometer method at different culture temperatures. The virulence of fungus BEdy1 to E. d. yunnanus was assessed on the lethality of the pests by treatment with different concentrations of spore suspension and estimated by comparison with the commercial fungal agent LH. Finally, the soybean leaves sprayed with BEdy1 spore suspension were used to evaluate the lethal effect after E. d. yunnanus feeding.【Result】The fungus caused the death of E. d. yunnanus under natural conditions was identified as Beauveria bassiana, named BEdy1. BEdy1 had a high growth rate at 22-28℃, and the highest growth rate was 4.34 mm·d-1 at 26℃. There was a high sporulation at 20-26℃, and the highest sporulation was 4.63×106 spores/mm2at 22℃. The mortality at 10th day of E. d. yunnanus adults treated with 1.0×105, 1.0×106, 1.0×107 and 1.0×108 spores/mL of B. bassiana BEdy1 spore suspension was 49.33%, 77.33%, 88.67% and 98.00%, respectively. The mortality of the control treatment was only 10%. The effect was best under the treatment of 1×108 spores/mL, with the LT50 of (6.79±0.13) d and the cadaver rate of 74.67%. The LC50 and LC95 at 10th day were 4.80×104 and 3.57×107 spores/mL, respectively. Treatment with 1×108 spores/mL of a commercial B. bassiana agent LH, the mortality and cadaver rate at 10th day were only 58.00% and 4.67%, which were significantly lower than those of the same concentration of BEdy1 treatment. After 10 days of E. d. yunnanus feeding on BEdy1 treated soybean leaves, the mortality and cadaver rate were 100% and 63.33%, respectively, and the LT50 was (5.27±0.35) d. 【Conclusion】Under appropriate conditions, the B. bassiana strain BEdy1 has fast growth rate, high sporulation, and high lethality rate to E. d. yunnanus adults, which has great potential for development and application in the future.

Key words: Ergania doriae yunnanus, Beauveria bassiana, biological characteristics, virulence, biological control

Fig. 1

The occurrence and harm of E. d. yunnanus A—F:The occurrence and harm in the field;G、H:The feeding damage of soybean leaf and pod after 2 days in the laboratory"

Fig. 2

Morphological characteristics and phylogenetic analysis of BEdy1 A:The cadaver of E. d. yunnanus infected with fungus BEdy1 in the field;B:Front of BEdy1 colony;C:Back of BEdy1 colony;D:Micrograph of conidia;E:Phylogenetic analysis。Beauveria caledonica;Beauveria vermiconia;Beauveria amorpha;Beauveria malawiensis;Beauveria brongniartii;Beauveria bassiana"

Table 1

Growth rate and sporulation of BEdy1 at different culture temperatures"

温度
Temperature (℃)
生长速率
Growth rate (mm·d-1)
产孢量
Sporulation (×106孢子/mm2)
16 2.09±0.10d 0.93±0.22cd
20 3.32±0.04c 2.95±0.35b
22 4.01±0.14b 4.63±0.63a
25 4.02±0.04b 3.63±0.30b
26 4.34±0.12a 3.55±0.35b
28 4.21±0.04ab 1.47±0.12c
30 2.01±0.02d 0.33±0.09de
35 0e 0e

Fig. 3

Virulence of B. bassiana BEdy1 to E. d. yunnanus with different spore concentrations"

Fig. 4

Pathogenesis of E. d. yunnanus treated with different spore concentrations of B. bassiana BEdy1 for 10 days"

Table 2

Virulence of B. bassiana BEdy1 to E. d. yunnanus at different times"

时间
Time (d)
毒力回归方程
Toxicity regression equation (y=)
LC50 (孢子/mL) LC95 (孢子/mL) 相关系数
Correlation coefficient
7 0.1287x-0.5047 6.43×107 2.02×1011 0.9470
8 0.2120x-0.8513 2.37×106 3.14×108 0.9879
9 0.1760x-0.4473 2.41×105 8.70×107 0.9560
10 0.1567x-0.2333 4.80×104 3.57×107 0.9196

Table 3

Virulence of commercial agent LH to E. d. yunnanus"

时间
Time (d)
死亡率
Mortality (%)
僵虫率
Cadaver rate (%)
LT50 (d)
8 38.67±4.81 *** 0 *** -
9 50.67±6.36 *** 2.00±1.15 *** -
10 58.00±3.06 *** 4.67±1.76 *** 9.00±0.52 **

Fig. 5

Virulence of spraying BEdy1 on leaves to E. d. yunnanus"

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