共生菌Arsenophonus、水稻品种和温度对褐飞虱黄绿绿僵菌发病率的影响

doi:10.16819/j.1001-7216.2017.7012

摘要/Abstract

摘要：

【目的】研究共生菌Arsenophonus、水稻和温度对褐飞虱黄绿绿僵菌发病率的影响,揭示影响褐飞虱黄绿绿僵菌发病的重要因子。【方法】通过共生菌Arsenophonus（感染和未感染）与水稻品种（TN1、IR56和Mudgo）以及Arsenophonus与温度（21℃、23℃、25℃、27℃、29℃和31℃）两个二因素完全随机区组试验,观察喷黄绿绿僵菌孢子悬浮液后不同时间褐飞虱黄绿绿僵菌的发病率。【结果】喷黄绿绿僵菌孢子悬浮液后,不同水稻品种上褐飞虱发病率均以含Arsenophonus的试虫较低,而不同温度下的褐飞虱发病率则仅喷菌处理后第3 d和5 d时在25℃、27℃和29℃时含Arsenophonus的试虫发病率显著较低,其他温度下无显著差异。双因素方差结果表明,Arsenophonus与水稻品种双因素试验中均以含Arsenophonus褐飞虱黄绿绿僵菌的发病率显著较低,而Arsenophonus与温度双因素试验中仅在喷菌处理后第3 d、5 d以含Arsenophonus褐飞虱黄绿绿僵菌的发病率显著较低。温度显著影响褐飞虱的发病率,23℃~29℃条件下褐飞虱的发病率较高,LT₅₀较短,为黄绿绿僵菌侵染褐飞虱的适温范围。其中,27℃时发病率最高,LT₅₀最短,最为适宜;31℃和21℃条件下褐飞虱发病率较低,LT₅₀较长,不利于黄绿绿僵菌对褐飞虱的侵染。温度还影响含Arsenophonus与不含Arsenophonus试虫LT₅₀的相对大小。其中,含Arsenophonus试虫LT₅₀在27℃、29℃时相对较长,在其他温度下则相同甚至较短。【结论】共生菌Arsenophonus显著抑制黄绿绿僵菌对褐飞虱的致病力,且该作用受到温度的显著影响;温度亦显著影响褐飞虱的黄绿绿僵菌发病率,而水稻品种以及Arsenophonus和水稻品种间、Arsenophonus和环境温度间的交互作用均无显著影响。

关键词: 褐飞虱, 黄绿绿僵菌, Arsenophonus, 温度, 水稻品种

Abstract:

【Objective】 To reveal the significant factors affecting the incidence rate of the brown planthopper (BPH) to Metarhizium flavoviride, laboratory bioassays were carried out to study the influence of symbiotic bacteria Arsenophonus, rice variety and temperature on the incidence rate of BPHs to M. flavoviride. 【Method】 Through two completely randomized block trial: 1) symbiotic bacteria Arsenophonus (infected and uninfected) versus rice varieties (TN1, IR56 and Mudgo), 2) and Arsenophonus (infected and uninfected) versus temperatures (21°C, 23°C, 25°C, 27°C, 29°C and 31°C), BPHs with symptoms of M. flavoviride infection were observed after exposure to the spore suspensions of M. flavoviride for different periods. 【Result】 For all tested rice varieties, the incidence rates of M. flavoviride-treated BPHs infected with Arsenophonus (BPH++) were lower than that of BPHs uninfected (BPH--). Out of tested temperatures, only under 25°C, 27°C, 29°C, BPH++ has lower incidence rates than BPH-- at 3 d and 5 d after spraying. However, there was no significant difference among others. The results of two-way ANOVA analysis between Arsenophonus and rice varieties also indicated that the incidence rate of BPH++ was lower than that of BPH--. Furthermore, two-way ANOVA results between Arsenophonus and temperatures indicated the incidence rate of BPH++ was lower than that of BPH-- at 3 d and 5 d after spraying. The effect of temperature was significant on the virulence of M. flavoviride to BPHs. BPHs had higher corrected incidence rates and shorter LT₅₀ under 23°C–29°C than that under 21°C or 31°C. Out of these BPHs, the highest corrected incidence rates and shortest LT₅₀ were found at 27°C. It suggested 23°C–29°C was suitable temperature for infection of M. flavoviride, among which 27°C was the best. This suggested that low or high temperatures would negatively affect the virulence of M. flavoviride to BPH. The temperature also affected the LT₅₀ of BPHs. The LT₅₀ of BPH++ was longer than that of BPH-- under 27°C or 29°C. However, the LT₅₀of BPH++ was equal or even shorter than that of BPH-- under other temperatures.【Conclusion】 Arsenophonus decreased the virulence of M. flavoviride to BPH, which was influenced by temperature. The temperature also affects the incidence rate of BPHs to Metarhizium flavoviride significantly. However, the effects of rice varieties and interactions of Arsenophonus and rice varieties and Arsenophonus and temperatures were not significant on the incidence rate.

Key words: Nilaparvata lugens, Metarhizium flavoviride, Arsenophonus, temperature, rice variety

中图分类号:

朱欢欢, 陈洋, 万品俊, 王渭霞, 赖凤香, 傅强. 共生菌Arsenophonus、水稻品种和温度对褐飞虱黄绿绿僵菌发病率的影响[J]. 中国水稻科学, 2017, 31(6): 643-651.

Huanhuan ZHU, Yang CHEN, Pinjun WAN, Weixia WANG, Fengxiang LAI, Qiang FU. Influence of Symbiotic Bacteria Arsenophonus, Rice Variety and Temperature on the Incidence Rate of Nilaparvata lugens to Metarhizium flavoviride[J]. Chinese Journal OF Rice Science, 2017, 31(6): 643-651.

图/表 5

图1 黄绿绿僵菌处理后三个水稻品种上含Arsenophonus与不含Arsenophonus褐飞虱发病率的比较 BPH++–含Arsenophonus的褐飞虱;BPH--–不含Arsenophonus的褐飞虱。同一品种不同试虫间,(*)、*、**分别表示在0.10、0.05、0.01水平上有显著差异,ns表示无显著差异(P>0.10)。

Fig. 1. Incidence rate of Metarhizium flavoviride-infected brown planthoppers with and without Arsenophonus reared on three rice varieties. BPH++, Brown planthopper with Arsenophonus; BPH--, Brown planthopper without Arsenophonus. The bar with (*),*,** above different test insects reared on the same rice variety after inoculation mean significant difference at the levels of 0.10, 0.05, 0.01, respectively, ns refers to no significant difference (P > 0.10).

图2 共生菌Arsenophonus(左)和水稻品种(右)对褐飞虱黄绿绿僵菌发病率的影响(平均值±标准误) BPH++–含Arsenophonus的褐飞虱;BPH--–不含Arsenophonus的褐飞虱。处理后同一时间内不同处理柱图上方具相同字母者示经邓肯新复极差法在5％水平下无显著差异。图4同。

Fig. 2. Effects of Arsenophonus (left) and rice varieties (right) on the incidence rate of Metarhizium flavoviride-infected brown planthoppers ( Mean ± SE ). BPH++, Brown planthopper with Arsenophonus; BPH--, Brown planthopper without Arsenophonus. The bars with a same letter for a same observation-day indicate no significant difference by Duncan's new multiple range method at P< 0.05. The same as in Figure 4.

图3 黄绿绿僵菌处理后在不同温度条件下含Arsenophonus与不含Arsenophonus褐飞虱发病率的比较同一温度不同试虫间*、**分别表示在显著水平0.05、0.01时有显著差异,ns表示无显著差异(P>0.05)。

Fig. 3. Incidence rate of Metarhizium flavoviride-infected brown planthoppers with and without Arsenophonus under different temperatures, respectively. The bar with *,** above different test insects under the same temperature after inoculation show significant difference at the levels of 0.05, 0.01, respectively; ns means no significant difference (P > 0.05).

图4 共生菌Arsenophonus (左)和温度(右)对褐飞虱黄绿绿僵菌校正发病率的影响(平均值±标准误)

Fig. 4. Effects of Arsenophonus (left) and temperatures (right) on the corrected incidence rate of Metarhizium flavoviride-infected brown planthoppers(Mean ± SE).

表1 不同温度下黄绿绿僵菌处理后褐飞虱的致死中时间LT50

Table 1 Median lethal time (LT50) of the brown planthoppers after Metarhizium flavoviride infestation at different temperatures. d

褐飞虱种群 Brown planthopper population	21℃	23℃	25℃	27℃	29℃	31℃
不含Arsenophonus Without Arsenophonus	11.4	7.6	5.6	4.7	5.3	>13.0
含Arsenophonus With Arsenophonus	10.2	7.0	5.6	5.3	6.4	8.8

参考文献 36

[1]	郭予元. 中国农作物病虫害.3版:上册. 北京: 中国农业出版社, 2015:93-104.
	Guo Y Y.Crop Diseases and Insect Pests in China. 3rd ed: Vol. I. Beijing: China Agriculture Press, 2015: 93-104. (in Chinese)
[2]	Douglas A E.Nutritional interactions in insect-microbial symbioses.Annu Rev Entomol, 1998, 43: 17-37.
[3]	Zchori-Fein E, Brown J K.Diversity of prokaryotes associated withBemisia tabaci (Gennadius), 2002, 95(6): 711-718.
[4]	Werren J H.Arsenophonus // Garrity T M. Bergey＇s Mannual of Systematic Bacteriology. New York: Springer Verlag, 2005.
[5]	Min S, Lee S W, Choi B R, Lee S H, Kwon D H.Insecticide resistance monitoring and correlation analysis to select appropriate insecticides againstNilaparvata lugens(Stål), a migratory pest in Korea. J Asia-Pacific Entomol, 2014, 17(4): 711-716.
[6]	王彦华, 王强, 沈晋良, 吴声敢, 俞瑞鲜, 赵学平, 苍涛, 吴长兴, 陈丽萍. 褐飞虱抗药性研究现状. 昆虫知识, 2009, 46(4): 518-524.
	Wang Y H, Wang Q, Shen J L, Wu S G, Yu R X, Zhao X P, Cang T, Wu C X, Chen L P.Current status of insecticide resistance in the brown planthopper,Nilaparvata lugens. Entomol Knowl, 2009, 46(4): 518-524. (in Chinese with English abstract)
[7]	Zhang X L, Liao X, Mao K K, Zhang K X, Wan H, Li J H.Insecticide resistance monitoring and correlation analysis of insecticides in field populations of the brown planthopperNilaparvata lugens(stål) in China 2012-2014. Pestic Biochem Physiol, 2016, 132: 13-20.
[8]	耿博闻. 黄绿绿僵菌防治稻飞虱的研究. 广州: 中山大学, 2004.
	Geng B W.Studies on biological control of rice planthopper with Metarrhizium anisopliae var. acridum. Guangzhou: Sun Yat-sen University, 2004. (in Chinese with English abstract)
[9]	徐阿妹. 黄绿绿僵菌培养生产技术及其在害虫防治中的应用. 合肥: 安徽农业大学, 2013.
	Xu A M.Culture production technology of Metarhizium flavoviride and its application in pest control. Hefei: Anhui Agricultural University, 2013. (in Chinese with English abstract)
[10]	Li M Y, Lin H F, Li S G, Xu A M, Feng M F.Efficiency of entomopathogenic fungi in the control of eggs of the brown planthopperNilaparvata lugens Stål (Homopera: Delphacidae). Afr J Microbiol Res, 2012, 6(44): 7162-7167.
[11]	常金梅, 何衍彪, 赵燕龙, 柳凤, 詹儒林. 绿僵菌致病力的制约因素研究. 安徽农业科学, 2010, 38(6): 2815-2816.
	Chang J M, He Y B, Zhao Y L, Liu F, Zhan R L.Study on the factors restraining the pathogeulcity ofMetarhizium sp. J Anhui Agric Sci, 2010, 38(6): 2815-2816. (in Chinese with English abstract)
[12]	岳梅, 雷仲仁, 孟涛, 王兆勇, 姚君明. 温度对绿僵菌LA菌株孢子萌发、酶活力及毒力的影响. 植物保护, 2010, 36(2): 56-60.
	Yue M, Lei Z R, Meng T, Wang Z Y, Yao J M.Temperature influence on conidial germination rate, enzyme activity and virulence ofMetarhizium anisopliae LA isolate. Plant Prot, 2010, 36(2): 56-60. (in Chinese with English abstract)
[13]	李佳颖, 许齐爱, 代晓彦, 汪郭凰, 任顺祥. 黄绿绿僵菌SM076菌株生物学特性研究. 广东农业科学, 2013 (9): 71-73.
	Li J Y, Xu Q A, Dai X Y, Wang G H, Ren S X.Study on the biological characteristics ofMetarhizium flavoviride. J. Guangdong Agric Sci, 2013(9): 71-73. (in Chinese with English abstract)
[14]	杜光祖, 杨卫秋, 周跃能, 和淑琪, 陈斌, 桂富荣, 李正跃. 黄绿绿僵菌KM1104对灰飞虱的毒力. 植物保护, 2015, 41(5): 207-211.
	Du G Z, Yang W Q, Zhou Y N, He S Q, Chen B, Gui F R, Li Z Y.Virulence ofMetarhizium flavoviride strain KM1104 against Laodelphax striatellus(Fallén). Plant Prot, 2015, 41(5): 207-211. (in Chinese with English abstract)
[15]	耿博闻, 张润杰. 低浓度唾嗓酮与黄绿绿僵菌对褐飞虱协同作用的生物测定. 植物保护学报, 2005, 32(1): 53-56.
	Geng W B, Zhang R J.Interactive effect of low concentration buprofezinMetarhizium anisopliae var. acridum on the mortalities of Nilaparvata legens(Hemiptera: Delphacidae) nymphs and adults in laboratory bioassays. Acta Phytophy Sin, 2005, 32(1): 53-56. (in Chinese with English abstract)
[16]	李茂业, 林华峰, 李世广, 金立. 黄绿绿僵菌Mf82菌株对不同虫态褐飞虱的毒力. 昆虫学报, 2012, 55(3): 316-323.
	Li M Y, Lin H F, Li S G, Jin L.Virulence ofMetarhizium flavoviride 82 to different developmental stages of the brown planthopper, Nilaparvata legens(Hemiptera: Delphacidae). Acta Entomol Sin, 2012, 55(3): 316-323. (in Chinese with English abstract)
[17]	陈宇, 陈洋, 王渭霞, 赖凤香, 傅强. 褐飞虱杀雄菌属共生菌Arsenophonus的传递方式与生物学意义初探. 中国水稻科学, 2014, 28(1): 92-96.
	Chen Y, Chen Y, Wang W X, Lai F X, Fu Q.A preliminary study on the transfer mode and biological significance of endosymbiontArsenophonus in Nilaparvata lugens. Chin J Rice Sci, 2014, 28(1): 92-96. (in Chinese with English abstract)
[18]	王渭霞, 罗举, 赖凤香, 傅强. 水稻褐飞虱内生共生细菌Arsenophonus的鉴定和系统分析. 昆虫学报, 2010, 53(6): 647-654.
	Wang W X, Luo J, Lai F X, Fu Q.Identification and phylogenetic analysis of symbiotic bacteriaArsenophonus from the rice brown planthopper, Nilaparvata lugens(Stål),Acta Entomol Sin 2010, 53(6): 647-654. (in Chinese with English abstract)
[19]	Wang Y, Cao L M, Zhang Y X, Cao C X, Liu F, Huang F K, Qiu Y F, Li Y B, Lou X J.Map-based cloning and characterization ofBPH29, a B3 domain-containing recessive gene conferring brown planthopper resistance in rice. J Exp Bot, 2015, 66(19): 6035-6045.
[20]	方中达. 植病研究方法. 北京:中国农业出版社, 1998: 40-72.
	Fang Z D.Methodology for plant pathology. Beijing: China Agriculture Press, 1998: 40-72. (in Chinese)
[21]	吴发红, 黄东益, 黄小龙, 周鑫, 程文杰. 几种真菌DNA提取方法的比较. 中国农学通报, 2009, 25(8): 62-64.
	Wu F H, Huang D Y, Huang X L, Zhou X, Cheng W J.Comparing study on several methods for DNA extraction from endophytic fungi.Chin Agric Sci Bull, 2009, 25(8): 62-64.
	( in Chinese with English abstract )
[22]	唐启义. DPS数据处理系统. 3版. 北京: 科学出版社, 2013.
	Tang Q Y.Applied Statistic Analysis and Date Processing System. 3ed. Beijing: Science Press, 2013. (in Chinese)
[23]	蒲蛰龙, 李增智. 昆虫真菌学. 合肥: 安徽科学技术出版社, 1996.
	Pu Z L, Li Z Z. Insect Mycology.Hefei: Anhui Science and Technology Press, 1996. (in Chinese)
[24]	刘爱英. 中国虫生真菌研究与应用. 北京: 中国农业科技出版社, 1991: 159-164.
	Liu A Y.Study and application of entomogenous fungi in China. Beijing: China Agriculture Science and Technology Press, 1991: 159-164. (in Chinese)
[25]	Mnyone L L, Kirby M J, Lwetoijera D W, Mpingwa M W, Simfukwe E T, Knols B G, Takken W, Russell T L.Tools for delivering entomopathogenic fungi to malaria mosquitoes: Effects of delivery surfaces on fungal efficacy and persistence.Malar J, 2010, 9: 246.
[26]	Jin S F, Feng M G, Chen J Q.Selection of globalMetarhizium isolates for the control of the rice pest Nilaparvata lugens(Homoptera: Delphacidae). Pest Manag Sci, 2008, 64(10): 1008-1014.
[27]	樊美珍, 郭超, 王姝清, 谢飙.不同菌株绿僵菌酯酶型与其致病力的关系. 西北林学院学报, 1990, 5(3): 1-6.
	Fan M Z, Guo C, Wang S Q, Xie B.Relathionship Between esterase type and pathogenic virulence of different strains ofMetarhizium spp. J Northwest For Coll, 1990, 5(3): 1-6. (in Chinese with English abstract)
[28]	孙召朋. 继代培养对球孢白僵菌毒力及毒力相关酶表达的影响. 长春: 吉林大学, 2011.
	Sun S P.Influence on the virulence of Beauveria bassiana and the expression of virulence related enzymes during subculturing. Changchun: Jilin University, 2011. (in Chinese with English abstract)
[29]	Scarborough C L, Ferrari J, Godfray H C J. Aphid protected from pathogen by endosymbiont.Science, 2005, 310(5755): 1781-1781.
[30]	汪敏捷, 刘强. 不同温湿度下绿僵菌对红缘天牛幼虫致病力的影响. 环境昆虫学报, 2014, 36(2): 151-156.
	Wang M J, Liu Q.Pathogenicity of theMetarhizium anisopliae to Asias halodendri larvae at different temperature and humidity conditions. J Eniron Entomol, 2014, 36(2): 151-156. (in Chinese with English abstract)
[31]	朱彬洲, 雷仲仁, 徐洪富, 王兆勇, 梁兴慧. 不同温湿度条件下绿僵菌LA菌株对东亚飞蝗的致病力. 中国生物防治, 2010, 26(4): 448-452.
	Zhu B Z, Lei Z R, Xu H F, Wang Z Y, Liang X H.Virulence ofMetarhizium anisopliae against Locusta migratoria manilensis under different temperatures and humidities. Chin J Biol Contr, 2010, 26(4): 448-452. (in Chinese with English abstract)
[32]	徐红星, 郑许松, 刘淑平, 叶恭银, 吕仲贤. 昆虫内共生菌在昆虫防御中的作用. 昆虫知识, 2009, 46(3): 350-354.
	Xu H X, Zheng X S, Liu S P, Ye G Y, Lu Z X.The role of endosymbionts in insect host resistance against adverse factors. Chin Bull Entomol, 2009, 46(3): 350-354. (in Chinese with English abstract)
[33]	Hansen A K, Jeong G, Paine T D, Stouthamer R.Frequency of secondary symbiont infection in an invasive psyllid relates to parasitism pressure on a geographic scale in California.App Environ Microb, 2007, 73(23): 7531-7535.
[34]	冯明光. 胞外蛋白酶和脂酶活性作为球孢白僵菌毒力指标的可靠性分析. 微生物学报, 1998(6): 461-467.
	Feng M G.Reliability of extracellular protease and lipase activities ofBeauveria bassiana isolates used as their virulence indices. Acta Microbiol Sin, 1998(6): 461-467. (in Chinese with English abstract)
[35]	林海萍, 魏锦瑜, 毛胜凤, 陈喜, 张立钦, 马良进. 球孢白僵菌蛋白酶、几丁质酶、脂肪酶活性与其毒力相关性. 中国生物防治, 2008, 24(3): 290-292.
	Lin H P, Wei J Y, Mao S F, Chen X, Zhang L Q, Ma L J.Correlation between protease, chitinase and lipase activities and virulence ofBeauveria bassiana against Monochamus alternatus. Chin J Biol Contr, 2008, 24(3): 290-292. (in Chinese with English abstract)
[36]	St Leger R J, Dumnds P K, Chamley A K, Cooper R M. Role of extracellular chymoelastase in the virulence ofMetarhizium anisopliae for Manduca sexta. J Invertebr Pathol, 1988, 52: 285-293.