制霉菌素对褐飞虱若虫解毒酶、尿酸酶含量的影响

doi:10.16819/j.1001-7216.2019.7129

摘要/Abstract

摘要：

【目的】为明确取食制霉菌素处理的水稻对褐飞虱[Nilaparvata lugens (Stål)]相关生理指标的影响,【方法】以不同浓度制霉菌素处理的水稻饲养褐飞虱,检测虫体内相关解毒酶含量与活性以及尿酸酶基因相对含量的变化。【结果】结果显示,取食制霉菌素处理的水稻,褐飞虱体内芳香基酰胺酶(aromatic amidase,AAD)含量前期下降,后期抑制作用逐渐消失,酶含量回升;虫体内O-脱乙基酶(O-deethyl enzyme,ODEE)含量在72 h的处理时长内显著低于对照;羧酸酯酶(carboxylesterase,CarE)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)的活性在不同浓度制霉菌素的处理下呈现不同的动态变化,高浓度制霉菌素(600 U/mL)处理的褐飞虱种群体内的CarE和GST的活性在处理时间段内均呈现诱导-抑制-诱导的现象,而中(300 U/mL)、低浓度(150 U/mL)处理种群中两种酶则随着处理时间的延长呈上升趋势。取食不同浓度制霉菌素处理水稻的褐飞虱种群体内类酵母共生菌尿酸酶基因在24 h、48 h之后与对照相比均无显著差异。取食48 h后,包括对照在内的4个处理尿酸酶基因相对转录水平均有下调趋势,当处理时长达72 h时,尿酸酶基因相对转录水平呈现上调趋势,但取食不同浓度制霉菌素处理水稻的褐飞虱种群体内含量显著低于对照。【结论】取食不同浓度制霉菌素处理的水稻对褐飞虱3龄若虫的4种主要的解毒酶具有不同的影响;制霉菌素处理可通过减少类酵母共生菌数量从而降低尿酸酶的含量。

关键词: 褐飞虱, 制霉菌素, 解毒酶, 尿酸酶基因

Abstract:

【Objective】To illuminate the effect of concentrations of nystatin on physiological indicators of brown planthoppers [Nilaparvata lugens (Stål)],【Method】detoxifying enzymes and uricase gene relative transcriptional level in brown planthoppers (Nilaparvata lugens) were measured after treatment with different concentrations of nystatin. 【Result】The content of aromatic amidase (AAD) in the brown planthopper(BPH) population fed on nystatin treated rice was decreased in the early stage, and increased gradually in the late stage. Nystatin treatment could significantly decrease the content of O-deethyl enzyme (ODEE). Activity of carboxylesterase(CarE) and glutathione S-transferase(GST) in BPH showed different trends under the treatment of nystatin. The activities of CarE and GST in 600 U/mL nystatin treated BPH population was induced-inhibited-induced during the treatment period, while there has been general uptrend of activities of two enzymes in the 300 U/mL and 150 U/mL nystatin treated populations within 72 h. The relative transcriptional level of uricase gene in different concentrations nystatin treated BPH populations had no significant difference with control by treatment with 24 h and 48 h. After feeding 48 h, the transcriptional level in three nystatin treated BPH populations and control population all decreased. To 72 h, the relative transcriptional level of uricase gene increased and transcriptional level in treated population was significantly lower than that in control population. 【Conclusion】Four detoxifying enzymes showed different trends after BPH was fed on rice treated with nystatin at different concentrations, nystatin treatment also reduce the content of uricase by reducing the number of yeast-like symbiotes.

Key words: Nilaparvata lugens, nystatin, detoxifying enzymes, uricase gene

中图分类号:

张珏锋, 李芳, 钟海英, 陈建明. 制霉菌素对褐飞虱若虫解毒酶、尿酸酶含量的影响[J]. 中国水稻科学, 2019, 33(2): 186-190.

Juefeng ZHANG, Fang LI, Haiying ZHONG, Jianming CHEN. Effects of Nystatin Treatment on Detoxification Enzymes and Uricase Content in Nymphs of the Brown Planthopper [Nilaparvata lugens (Stål)][J]. Chinese Journal OF Rice Science, 2019, 33(2): 186-190.

图/表 2

图1 制霉菌素对褐飞虱解毒酶的影响 A–芳基酰胺酶(AAD);B–O-脱乙基酶(ODEE);C–羧酸酯酶(CarE);D–谷胱甘肽S-转移酶(GST)。图中数据为3次独立重复的平均值±标准差,同一时间点不同处理间标注不同小写字母表示在P=0.05水平上差异显著。

Fig. 1. Effect of nystatin on detoxifying enzymes in the Nilaparvata lugens. A, Aromatic a midase(AAD); B, O-deethyl enzyme(ODEE); C, Carboxyle sterase(CarE); D, Glutathione S-transterase(GST). Data are shown as mean±SD. The data at the same time point followed with different lowercase letters are significantly different at 0.05 level.

表1 取食不同处理水稻后褐飞虱体内尿酸酶基因相对转录水平

Table 1 Changes in uricase relative transcriptional level in N. lugens after nystatin treatment.

制霉菌素浓度 Nystatin concentration /(U·L^-1)	取样时间 Sampling time/h
制霉菌素浓度 Nystatin concentration /(U·L^-1)	24	48	72
0(CK)	1.99±0.59 aA	0.60±0.11 aA	17.53±2.31 aA
150	3.02±0.84 aA	0.99±0.07 aA	1.62±0.25 cB
300	2.36±0.55 aA	1.57±0.68 aA	9.77±2.70 bB
600	4.31±1.11 aA	1.31±0.60 aA	2.34±0.63 cB

参考文献 20

[1]	Tokuda G, Elbourne L D H, Kinjo Y, Saitoh S, Sabree Z, Hojo M, Yamada A, Hayashi Y, Shigenobu S, Bandi C, Paulsen I T, Watanabe H, Lo N. Maintenance of essential amino acid synthesis pathways in the Blattabacterium cuenoti symbiont of a wood-feeding cockroach. Biol Lett, 2013, 9(3): 20121153.
[2]	Thompson B M, Grebenok R J, Behmer S T, Gruner D S.Microbial symbionts shape the sterol profile of the xylem-feeding woodwasp, Sirex noctilio. J Chem Ecol, 2013, 39(1): 129-139.
[3]	Price D R G, Feng H, Baker J D, Bavan S, Luetje C W, Wilson A C. Aphid amino acid transporter regulates glutamine supply to intracellular bacterial symbionts. PNAS, 2014, 111(1): 320-325.
[4]	Martín M J, Calpena A C, Fernández F, Mallandrich M, Galvez P, Clares B.Development of alginate micros pheres as nystatin carriers for oral mucosa drug delivery.Carbohyd Poly, 2015, 117: 140-149.
[5]	张珏锋, 何月平, 陈建明. 制霉菌素对褐飞虱取食行为, 营养及共生菌数量的影响. 植物保护学报, 2015 (2): 264-270.
	Zhang J F, He Y P, Chen J M.Effects of nystatin on the feeding behavior, nutrition and yeast-like symbiont number of the brown planthopper, Nilaparvata lugens(Stål). J Plant Prot, 2015, 42(2): 264-270. (in Chinese with English abstract)
[6]	王国超. 褐飞虱体内类酵母共生菌的营养功能及其分子基础的研究.北京: 中国农业科学院, 2005.
	Wang G C. The nutritional function and its molecular basis of yeast like symbiotes in rice brown planthopper Nilaparvata lugens(Stål). Beijing: Chinese Academy of Agricultural Sciences.
[7]	Bao H B, Gao H L, Zhang Y X, Fan D Z, Fang J C, Liu Z W.The roles of CYP6AY1 and CYP6ER1 in imidacloprid resistance in the brown planthopper: Expression levels and detoxification efficiency.Pestic Biochem Physiol, 2016, 129: 70-74.
[8]	Zhang X L, Liao X, Mao K K, Yang P, Li D Y, Alia E, Wan H, Li J H.The role of detoxifying enzymes in field-evolved resistance to nitenpyram in the brown planthopperNilaparvata lugens in China. Crop Prot, 2017, 94: 106-114.
[9]	Liu N N, Zhu F, Xu Q,. Pridgeon J W, Gao X W.Behavioral change, physiological modification, and metabolic detoxification: Mechanisms of insecticide resistance.Acta Entomol Sin, 2006, 49(4): 671-679.
[10]	邱星辉. P细胞色素450 介导的昆虫抗药性的分子机制. 昆虫学报, 2014, 57(4): 477-482.
	Qiu X H.Molecular mechanissms of insecticide resistance mediated by cytochrome P450s in insects.Acta Entomol Sin, 2014, 57(4): 477-482.(in Chinese with English abstract)
[11]	孙璟琰, 梁沛, 高希武. 小菜蛾芳基酰胺酶表达及其与呋喃虫酰肼抗性的关系. 应用昆虫学报, 2012, 49(2): 396-402.
	Sun J Y, Liang P, Gao X W.The distribution of aryl-acylamidase in the diamondback moth, Plutella xylostella and its relationship to fufenozide resistance. Chin J Appl Entomol, 2012, 49(2): 396-402.(in Chinese with English abstract)
[12]	邢静, 梁沛, 高希武. 亚致死浓度氯虫苯甲酰胺对小菜蛾药剂敏感度和解毒酶活性的影响. 农药学学报, 2011, 13(5): 464-470.
	Xing J, Liang P, Gao X W.Effects of sublethal concentrations of chlorantraniliprole on insecticide susceptibility and detoxifying enzyme activity in Plutella xylostella.Chin J Pestic Sci, 2011, 13(5): 464-470. (in Chinese)
[13]	范银君, 康志娇, 王志超, 史雪岩,高希武. 茉莉酸甲酯处理对棉铃虫生长和解毒能力的影响. 昆虫学报, 2014, 3: 007.
	Fan Y J, Kang Z J, Wang Z C, Shi X Y, Gao X W.Effects of methyl jasmonate treatment on the growth and detoxifyingabilities of Helicoverpa armigera(Lepid optera: Noctuidae). Acta Entomol Sin, 2014, 57(3): 315-322. (in Chinese with English abstract)
[14]	李阳, 史雪岩, 高希武. 增效醚对二化螟幼虫 7-乙氧基香豆素-O-脱乙基酶和羧酸酯酶活性的影响. 昆虫学报, 2016, 59(11): 1159-1165.
	Li Y, Shi X Y, Gao X W. In vivo effects of piperonyl butoxide on the activities of 7-ethoxycoumarin O- deethylase and carboxylesterase in larval Chilo sup ressalis(Lepidoptera: Pyralidae). Acta Entomol Sin, 2016, 59(11): 1159-1165. (in Chinese with English abstract)
[15]	Cao G C, Lu Q,Zhang L L, Guo F, Liang G M, Wu K M, Wyckhuys K A G, Guo Y Y. Toxicity of chlorantraniliproleto Cry1Ac-susceptible andresistant strains ofHelicoverpa armigera.Pestic BiochemPhysiol, 2010, 98(1): 99-103.
[16]	Campanhola C, Plapp F W.Pyrethroid resistance in the tobacco budworm (Lepidoptera: Noctuidae): Insecticide bioassays and field monitoring.J Econ Entomol, 1989, 82(1): 22-28.
[17]	吴青君, 张文吉, 张友军, 徐宝云, 朱国荣. 表皮穿透和 GABAA 受体不敏感性在小菜蛾对阿维菌素抗性中的作用. 昆虫学报, 2002, 45(3): 336-340.
	Wu Q J, Zhang W J, Zhang Y J, Xu B Y, Zhu G R.Cuticular penetration and desensitivity of GABA Areceptor in abamectin resistant Plutella xylostella L. Acta Entomol Sin, 2002, 45(3): 336-340.(in Chinese with English abstract)
[18]	Yu S J, Nguyen S N, Abo-elghar G E. Biochemical characteristics of insecticide resistance in the fall armyworm, Spodoptera frugiperda(JE Smith). Pestic Biochem Physiol, 2003, 77(1): 1-11.
[19]	Masson P, Froment M T, Gillon E, Nachon F, Darvesh S, Schopfer L M.Kinetic analysis of butyrylcholinesterase -catalyzed hydrolysis of acetanilides.Biochim Biophy Acta, 2007, 1774(9): 1139-1147.
[20]	Chen Z Y, Wang Z Y, He X P, Guo X N, Li W W, Zhang B R.Uricase production by a recombinant Hansenula polymorpha strain harboring Candida utilis uricase gene.Appl Microbiol Biotechnol, 2008, 79(4): 545-554.