布氏白僵菌菌株JG-17的鉴定及对3种金龟子的毒力测定

doi:10.3864/j.issn.0578-1752.2024.12.008

Abstract

Abstract:

【Objective】 Based on the current situation of the wide variety and serious damage of scarabs, and the urgent demand of agriculture for biological control with eco-friendly and sustainable management, the virulence of Beauveria brongniartii strain JG-17 to the larvae and adults of three important scarabs in China was studied in order to provide excellent microbial resources for the research and development of scarabs biological control agents.【Method】A high virulence fungus strain was isolated from the infected adult scarab cadavers by spore dilution method and re-inoculation experiment. The taxonomic status of strain JG-17 was determined by morphology and rDNA-ITS gene sequence analysis. The growth rate and sporulation of strain JG-17 at different culture temperatures were measured by cross method and hemocytometer method. The virulence of strain JG-17 to the adults and larvae of Holotrichia oblita, Holotrichia parallela and Anomala corpulenta was determined by immersion method and the control efficacy against the 2nd instar larvae of H. parallela was determined by peanut pot experiment.【Result】A strain of Beauveria JG-17, which caused high virulence against adults and larvae of three scarabs, was isolated from the naturally infected scarabs adult. Strain JG-17 was identified as B. brongniartii combined with the morphological and molecular identification results. 25-29 ℃ is the optimal growth temperature for strain JG-17, and the growth rate is greater than 4 mm·d^-1. The growth rate and sporulation of strain JG-17 are the highest at 27 ℃. The cumulative mortality at 13 d of the adults of H. oblita, A. corpulenta and H. parallela treated with 1×10⁸ spores/mL of strain JG-17 spore suspension was 78.0%, 100.0% and 86.0% with the LT₅₀ values of 9.458, 6.911 and 8.391 d, respectively. At the same treatment concentration, the virulence of JG-17 to the larvae of H. parallela was higher than that of A. corpulenta and H. oblita. At the 1×10⁸ spores/mL of JG-17 concentration treatment, the cumulative mortality at 13 d of the 1st instar larvae of H. oblita, A. corpulenta and H. parallela was 76.7%, 96.7% and 100.0%, respectively, and that of the 2nd instar larvae was 66.7%, 36.7% and 100.0%, respectively. Under different spore concentrations of JG-17 treatment, the LT₅₀ of JG-17 against the 1st and 2nd instar larvae of H. parallela was significantly lower than that of A. corpulenta and H. oblita. In the pot experiment, there was no significant difference in larvae mortality between spraying evenly around the root and the furrow application with same spore dosage during the inter-tillage period, and more than 10 times of spore were needed to achieve similar control efficacy with spreading application before sowing.【Conclusion】B. brongniartii JG-17 has a high lethality rate to the adults and larvae of H. oblita, H. parallela and A. corpulenta. It is a biocontrol microbial resource with great potential for development and application to control underground scarab pests.

Key words: Beauveria brongniartii, scarab beetle, larva, adult, isolation and identification, virulence

CAO WeiPing, FENG Shuo, CHENG JiaXu, CHEN Dan, WU QingJun, SONG Jian. Species Identification of Beauveria brongniartii Strain JG-17 and Virulence Against Three Scarabaeoidae Pests[J].Scientia Agricultura Sinica, 2024, 57(12): 2364-2377.

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菌株 Isolate	采集地 Collecting site	菌落形态 Colony texture	菌落颜色 Colony color	生长速率 Growth rate (mm·d^-1)	产孢量 Sporulation (×10⁶spores/cm²)	侵染率 Infection rate (%)
JG-4	河北保定Baoding, Hebei	毡状Felt-like	白色White	3.65±0.16d	4.83±0.93c	32.0±4.5cd
JG-5	河北博野Boye, Hebei	绒状Villous-like	白色White	5.39±0.15a	7.65±0.76a	18.0±8.4e
JG-9	河北沧州Cangzhou, Hebei	绒状Villous-like	黄色Yellow	2.64±0.13f	3.24±0.50d	20.0±7.1e
JG-12	河北高阳Gaoyang, Hebei	毡状Felt-like	淡黄色Light yellow	4.07±0.04c	4.98±0.56c	26.0±5.5de
JG-17	河北清苑Qingyuan, Hebei	绒状Villous-like	乳白略黄Yellowish white	4.23±0.07b	7.29±0.65a	92.0±8.4a
JG-18	河北满城Mancheng, Hebei	绒状Villous-like	淡黄色Light yellow	3.31±0.04e	3.43±0.60d	46.0±8.9b
JG-21	河北乐亭Laoting, Hebei	绒状Villous-like	乳白色Cream white	4.02±0.10c	6.31±0.48b	40.0±10.0bc

温度 Temperature (℃)	生长速率 Growth rate (mm·d^-1)	产孢量Sporulation (×10⁶ spores/cm²)
18	1.64±0.05e	2.41±0.34f
21	2.74±0.05c	3.50±0.71e
23	3.70±0.12b	5.17±0.68d
25	4.12±0.13a	6.33±0.49bc
27	4.23±0.07a	7.66±0.73a
29	4.16±0.11a	6.96±0.63ab
31	2.28±0.08d	5.78±0.63cd
33	1.38±0.08f	2.90±0.84ef
35	0g	0g

金龟子种类 Scarab species	僵虫率Cadaver rate (%)
金龟子种类 Scarab species	1×10⁸ spores/mL	5×10⁷spores/mL	2×10⁷ spores/mL	5×10⁶ spores/mL	2×10⁶ spores/mL
华北大黑鳃金龟H. oblita	72.0±4.5c	56.0±5.5c	48.0±8.4c	28.0±8.4c	12.0±4.5c
铜绿丽金龟A. corpulenta	94.0±5.5a	86.0±5.5a	78.0±4.5a	66.0±5.5a	58.0±4.5a
暗黑鳃金龟H. parallela	82.0±4.5b	70.0±7.1b	62.0±4.5b	52.0±4.5b	30.0±7.1b

金龟子种类 Scarab species	孢子浓度 Spore concentration (spores/mL)	回归方程 Regression equation (y=)	LT₅₀(d)，95%置信限 Confidence limit	χ²	P
华北大黑鳃金龟 H. oblita	1×10⁸	-2.666+0.282x	9.458 (8.834-10.107)	6.075	1.000
华北大黑鳃金龟 H. oblita	5×10⁷	-2.751+0.249x	11.033 (10.311-11.968)	5.790	1.000
铜绿丽金龟 A. corpulenta	1×10⁸	-2.783+0.403x	6.911 (6.330-7.410)	6.732	1.000
铜绿丽金龟 A. corpulenta	5×10⁷	-2.431+0.314x	7.846 (7.192-8.426)	11.861	0.973
暗黑鳃金龟 H. parallela	1×10⁸	-2.066+0.246x	8.391 (7.638-9.084)	6.034	1.000
暗黑鳃金龟 H. parallela	5×10⁷	-2.241+0.227x	9.875 (9.134-10.715)	7.490	0.999

金龟子种类 Scarab species	龄期 Stage	累计死亡率Cumulative mortality (%)
金龟子种类 Scarab species	龄期 Stage	1×10⁸ spores/mL	5×10⁷ spores/mL	2×10⁷ spores/mL	5×10⁶ spores/mL	2×10⁶ spores/mL	CK
华北大黑鳃金龟 H. oblita	1龄1st instar	76.7±11.5	63.3±5.8	50.0±10.0	23.3±15.3	6.7±5.8	0
华北大黑鳃金龟 H. oblita	2龄2nd instar	66.7±5.8	53.3±5.8	33.3±6.7	13.3±3.3	3.3±5.8	0
铜绿丽金龟 A. corpulenta	1龄1st instar	96.7±5.8	90.0±10.0	76.7±5.8	40.0±10.0	26.7±5.8	6.7±5.8
铜绿丽金龟 A. corpulenta	2龄2nd instar	36.7±5.8	23.3±5.8	13.3±5.8	6.7±5.8	0	0
暗黑鳃金龟 H. parallela	1龄1st instar	100.0±0	100.0±0	96.7±5.8	86.7±5.8	60.0±0	0
暗黑鳃金龟 H. parallela	2龄2nd instar	100.0±0	93.3±5.8	76.7±5.8	53.3±5.8	36.7±5.8	0

Species Identification of Beauveria brongniartii Strain JG-17 and Virulence Against Three Scarabaeoidae Pests

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处理Treatment	死亡率Mortality (%)	僵虫率Cadaver rate (%)
撒施Spreading application (500×10⁸spores/pot)	50.00±0e	41.67±7.22c
撒施Spreading application (1000×10⁸spores/pot)	66.67±7.22cd	58.33±7.22ab
撒施Spreading application (2000×10⁸spores/pot)	83.33±7.22ab	70.83±7.22a
淋灌Spraying around peanut root (20×10⁸spores/pot)	45.83±7.22e	37.50±12.50c
淋灌Spraying around peanut root (40×10⁸spores/pot)	58.33±7.22de	50.00±0bc
淋灌Spraying around peanut root (80×10⁸spores/pot)	70.83±7.22bcd	66.67±7.22a
沟施Furrow application (40×10⁸spores/pot)	50.00±0e	41.67±7.22c
沟施Furrow application (80×10⁸spores/pot)	66.67±7.22cd	62.50±0ab
沟施Furrow application (160×10⁸spores/pot)	75.00±12.50abc	70.83±7.22a
45%毒死蜱Chlorpyrifos (2000×)	87.50±12.50a	0d
CK	8.33±7.22f	0d

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