楊靜美 張志祥 葉滔 蔡奕浩 陳趣 王文凱
摘要:【目的】研究曼陀羅不同部位提取物對(duì)小、中型紅火蟻工蟻的毒殺活性及行為作用,為利用植物源藥劑防治紅火蟻及其他有害生物提供科學(xué)依據(jù)。【方法】采用水試管喂養(yǎng)法,探究曼陀羅根、莖、葉、花和種子提取物對(duì)紅火蟻的毒力及攀爬率、抓附率、行走率和聚集率等行為的影響。【結(jié)果】曼陀羅不同部位提取物對(duì)小、中型紅火蟻均具有一定的毒力,且隨處理時(shí)間的延長(zhǎng)紅火蟻死亡率上升,其中以曼陀羅干葉和干花對(duì)兩種蟻型紅火蟻的毒殺效果最好,處理11 d后,干葉和干花提取物處理小型蟻的死亡率分別為98.67%和100.00%、中型蟻的死亡率均為100.00%。曼陀羅不同部位提取物對(duì)紅火蟻的抓附率、攀爬率、聚集率和行走率等行為也具有抑制作用,其中以曼陀羅干花和干葉的作用效果最好,處理11 d后,干花提取物處理兩種蟻型紅火蟻的攀爬率、抓附率、聚集率和行走率均為0;干葉提取物處理小型蟻的攀爬率、抓附率和行走率均為1.33%、聚集率為0,而中型蟻的攀爬率、抓附率、行走率和聚集率均為0?!窘Y(jié)論】曼陀羅不同部位提取物對(duì)兩種蟻型紅火蟻均具有良好的毒殺活性,對(duì)其行為有一定的抑制作用,其中以干花的影響力最明顯,具有開(kāi)發(fā)成新型綠色環(huán)保紅火蟻防治植物材料的潛力。
關(guān)鍵詞: 紅火蟻;曼陀羅;毒殺活性;行為抑制作用
中圖分類(lèi)號(hào): S433; S759.82? ? ? ? ? ? ? ? ? ? ? ?文獻(xiàn)標(biāo)志碼: A 文章編號(hào):2095-1191(2019)11-2475-06
Effects of different parts of Datura stramonium on the
Solenopsis invicta Buren workers
YANG Jing-mei1,2, ZHANG Zhi-xiang3*, YE Tao4, CAI Yi-hao2,
CHEN Qu2, WANG Wen-kai1*
(1College of Agriculture, Yangtze University, Jingzhou, Hubei? 434025, China; 2College of Agriculture and Biology, Zhongkai University of Agriculture and Engineering, Guangzhou? 510225, China; 3College of Agriculture, South China Agricultural University/Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, Guangzhou 510642, China; 4School of Pharmaceutical Sciences, Guangzhou Medical University, Guangzhou? 511436, China)
Abstract:【Objective】The insecticidal activity and the behavior of medium and small-sized workers of red imported fire ants were studied with Datura stramonium extracts, providing the basis for the control of red imported fire ants and other pest by botanical insecticides. 【Method】The effects of extracts from root, stem, leaf, flower and seeds of D. stramonium on the toxicity, climbing rate, grasping rate, walking rate and aggregation rate of red fire ants were studied by aqueous solution tubes method. 【Result】Extracts from different parts of D. stramonium had toxicity to both small and medium-sized red imported fire ants, and the mortality of red imported fire ants increased with time. Among them, dried leaves and dried flowers of D. stramonium had the best toxicity on the two types of red imported fire ants. After 11 d, the mortality of small ants treated with dried leaves and flowers were 98.67% and 100.00%, respectively, and the mortality of medium ants were both 100.00%. Extracts from different parts of D. stramonium also had inhibitory effects on the grasping rate, climbing rate, aggregation rate and walking rate of red imported fire ants, the effects of dried flowers and leaves were the best. After 11 d, the climbing rate, grasping rate, aggregation rate and walking rate treated with dried flower extracts for the two ant types were all 0. The climbing rate, grasping rate and walking rate of small ants treated by dried leaves were all 1.33%, and aggregation rate was 0, while the climbing rate, grasping rate, walking rate and aggregation rate of medium ants were 0 with dry leaves. 【Conclusion】All the parts of D. stramonium have insecticidal activity and behavioral inhibition effects on two types of the red imported fire ants, and dries flower has the optimal effects, may be the green environmental protection agent to control red imported fire ants.
Key words: Solenopsis invicta Buren; Datura stramonium; insecticidal activity; behavioral inhibition effect
0 引言
【研究意義】紅火蟻(Solenopsis invicta Buren,red imported fire ant,RIFA)是世界上最嚴(yán)重的100種外來(lái)入侵物種之一,是一種營(yíng)社會(huì)生活的昆蟲(chóng),原產(chǎn)于南美洲,后迅速擴(kuò)散到澳大利亞、新西蘭、馬來(lái)西亞、泰國(guó)和菲律賓等地(Zhang et al.,2007;趙瑾等,2009;Wang and Lu,2017)。我國(guó)于2004年首次發(fā)現(xiàn)紅火蟻,后在南方多個(gè)省份發(fā)生危害,其對(duì)生態(tài)環(huán)境、人類(lèi)、動(dòng)植物及公共設(shè)施等造成了嚴(yán)重危害,甚至危及人類(lèi)的生命(Ascunce et al.,2011;Tian et al.,2015;吳偉斌等,2017;王智楠等,2019)。目前紅火蟻的防治方法多集中在化學(xué)、物理和生物方面,其中化學(xué)防治的效果最好,但化學(xué)防治給環(huán)境帶來(lái)的潛在危害不容忽視,因此,尋求有效的綠色藥劑是當(dāng)前紅火蟻防治的重要目標(biāo)?!厩叭搜芯窟M(jìn)展】紅火蟻綠色藥劑防治主要集中在運(yùn)用植物源藥劑和生物防治上。研究發(fā)現(xiàn),多種植物源藥劑對(duì)紅火蟻具有一定的防治效果。Li等(2014)研究發(fā)現(xiàn),非洲山毛豆豆莢和枝葉中的揮發(fā)性物質(zhì)蒎烯等對(duì)紅火蟻有一定的熏蒸毒力,作用1~12 h后小型紅火蟻的死亡率分別為18.33%~100.00%和13.33%~100.00%。Zhang等(2014)的研究結(jié)果表明,青蒿油有較強(qiáng)的殺紅火蟻活性,且在密閉環(huán)境中抑制作用更強(qiáng)。新鮮九里香葉、落葉和干葉提取物作用1~9 d后小型紅火蟻的死亡率分別為5.00%~100.00%、11.67%~93.33%和15.00%~83.33%(Huang et al.,2016a);作用48 h后,新月蕨甲醇和乙酸乙酯提取物對(duì)大型紅火蟻的LC50分別為524.0和145.9 μg/g,對(duì)小型紅火蟻的LC50分別為321.9和90.0 μg/g(Huang et al.,2016b)。王磊等(2018)研究發(fā)現(xiàn),1×108孢子/mL濃度的金龜子綠僵菌M09處理10 d后對(duì)紅火蟻幼蟲(chóng)的侵染率為98.9%,蛹為100.0%,有較強(qiáng)的致病力,致死速度較快。白花曼陀羅(Datura metel Linn.)為茄科(Solanacea)曼陀羅屬(Datura Linn.)植物,又名洋金花,分布廣泛,內(nèi)含莨菪堿和東莨菪堿等,是常見(jiàn)的中草藥及殺蟲(chóng)植物(張宏利等,2004;陸秀紅等,2006)。研究發(fā)現(xiàn),白花曼陀羅種子提取液對(duì)線蟲(chóng)有較好的毒殺作用(Goswami and Vijayalakshmi,1987;Gamal et al.,2008);曼陀羅葉、籽提取物對(duì)立枯絲核菌菌絲、甘藍(lán)蚜蟲(chóng)和朱砂葉螨等均具有良好的生物活性(楊文,2006)?!颈狙芯壳腥朦c(diǎn)】基于植物源藥劑對(duì)紅火蟻的研究已有較多報(bào)道,但曼陀羅對(duì)紅火蟻的作用研究鮮見(jiàn)報(bào)道?!緮M解決的關(guān)鍵問(wèn)題】以白花曼陀羅植物為材料,通過(guò)研缽研磨,超聲波振動(dòng)提取其根、莖、葉、花和種子中的有毒物質(zhì),采用水試管喂毒法,探究各提取物對(duì)小、中型紅火蟻工蟻的毒力及行走率、抓附率、攀爬率和聚集率等行為的影響,為利用植物源藥劑防治紅火蟻及其他有害生物提供科學(xué)依據(jù)。
1 材料與方法
1. 1 試驗(yàn)材料
1. 1. 1 供試植物材料 白花曼陀羅,產(chǎn)自江蘇徐州。
1. 1. 2 供試紅火蟻工蟻 紅火蟻(S. invicta)工蟻來(lái)源于華南農(nóng)業(yè)大學(xué)附近農(nóng)場(chǎng)。小型蟻頭寬0.6~0.7 mm,中型蟻頭寬0.7~0.9 mm。將紅火蟻工蟻連巢穴裝入桶壁有滑石粉的塑料桶中,室內(nèi)養(yǎng)殖(溫度25~28 ℃,濕度70%~80%,光照∶黑暗=12 h∶12 h,桶中放入水試管、火腿腸飼養(yǎng))1周后待用。
1. 2 試驗(yàn)方法
1. 2. 1 曼陀羅各部位有效成分提取 分別將曼陀羅的鮮根、鮮莖、鮮葉、干葉、干花和種子加入適量乙醇和蒸餾水充分研磨,超聲波1 h后放入烘箱中50 ℃下烘干(尚天翠,2016)。取出后各加入適量乙醇和蒸餾水?dāng)嚢枵袷?,得到濃度分別為鮮莖1.5 g/45 mL和1.5 g/30 mL、鮮根1.0 g/40 mL、鮮葉1.5 g/45 mL和1.5 g/30 mL、干葉1.5 g/90 mL和1.5 g/75 mL、干花1.5 g/90 mL和1.5 g/75 mL、種子1.5 g/45 mL和1.5 g/30 mL的溶液。對(duì)照組僅在蒸餾水中加入乙醇。試驗(yàn)組和對(duì)照組均將乙醇揮發(fā)至無(wú)氣味。
1. 2. 2 曼陀羅各部位提取物對(duì)紅火蟻的毒殺活性測(cè)定 采用試管中加入溶液,管口塞入棉花致溶液緩慢滲出的方式喂養(yǎng)紅火蟻,并進(jìn)行毒力測(cè)定(Zhang et al.,2017)。將不同部位的曼陀羅溶液(小型蟻:鮮莖1.5 g/45 mL、鮮根1.0 g/40 mL、鮮葉1.5 g/45 mL、干葉1.5 g/90 mL、干花1.5 g/90 mL、種子1.5 g/45 mL;中型蟻:鮮莖1.5 g/30 mL、鮮根1.0 g/40 mL、鮮葉1.5 g/30 mL、干葉1.5 g/75 mL、干花1.5 g/75 mL、種子1.5 g/30 mL)依次加入2 mL試管中,試管口塞入棉花后放入塑料杯中,杯口以下1~2 cm處涂抹滑石粉。在每個(gè)塑料杯中接入25只紅火蟻,每處理重復(fù)3次,對(duì)照則在試管中加入蒸餾水。每個(gè)塑料杯中放入少許火腿腸喂食紅火蟻,下同。喂食1、3、5、7、9和11 d后,觀察曼陀羅不同部位提取液及濃度下紅火蟻的死亡情況。根據(jù)公式計(jì)算紅火蟻死亡率平均值:死亡率(%)=死亡工蟻數(shù)/總工蟻數(shù)×100。
1. 2. 3 曼陀羅各部位提取物對(duì)紅火蟻?zhàn)ジ铰?、攀爬率、聚集率和行走率的影?按照1.2.2的方法,觀察杯中紅火蟻的行為。攀爬率測(cè)定:將竹簽從底部開(kāi)始每隔1 cm處標(biāo)記,輕輕碰觸杯中的紅火蟻至竹簽上,其在竹簽上攀爬的距離超過(guò)3 cm時(shí),認(rèn)為紅火蟻具有攀爬能力(Zhang et al.,2013);計(jì)算攀爬率:攀爬率(%)=有攀爬能力的工蟻數(shù)/總工蟻數(shù)×100。抓附率測(cè)定:輕輕振動(dòng)塑料杯,待紅火蟻分布在杯底靜置10 s后,將白紙緊貼于杯口,待杯子翻轉(zhuǎn)180°后,停頓5 s,隨后翻轉(zhuǎn)杯子至原來(lái)位置,觀察杯中余下的紅火蟻數(shù)量,掉落在杯中的紅火蟻即為不具有抓附能力的螞蟻;計(jì)算抓附率:抓附率(%) =(總工蟻數(shù)-杯中工蟻數(shù))/總工蟻數(shù)×100。聚集率測(cè)定:將塑料杯輕輕振動(dòng),待紅火蟻在杯底靜置10 s后觀察紅火蟻的聚集情況,3只及以上能聚集在一起即為有聚集能力;計(jì)算聚集率:聚集率(%)=聚集的工蟻數(shù)/總工蟻數(shù)×100。行走率測(cè)定:在A4白紙上打印多個(gè)1 cm×1 cm的小正方形,將紅火蟻放到白紙上,若紅火蟻3 s內(nèi)能行走3 cm,則認(rèn)為其具有行走能力;計(jì)算行走率:行走率(%)=具有行走能力的工蟻數(shù)/總工蟻數(shù)×100。
1. 3 統(tǒng)計(jì)分析
利用SAS 9.0對(duì)試驗(yàn)數(shù)據(jù)進(jìn)行方差分析。
2 結(jié)果與分析
2. 1 曼陀羅對(duì)紅火蟻的毒力作用測(cè)定結(jié)果
曼陀羅不同部位提取物對(duì)小、中型紅火蟻的毒力測(cè)定結(jié)果(表1)顯示,在不同曼陀羅部位提取物作用下紅火蟻的死亡率均隨處理時(shí)間的延長(zhǎng)而上升,其中,曼陀羅干葉和干花對(duì)兩種紅火蟻工蟻的毒殺效果最好,處理11 d后,干葉和干花提取物處理小型蟻的死亡率分別為98.67%和100.00%,其他部位提取物處理小型蟻的死亡率為65.33%~74.67%,與對(duì)照(32.00%)相比均達(dá)顯著差異水平(P<0.05,下同);干葉和干花提取物處理中型蟻的死亡率均為100.00%,其他部位提取物處理中型蟻的死亡率為40.00%~90.67%,與對(duì)照(6.67%)相比均達(dá)顯著差異水平。
2. 2 曼陀羅對(duì)紅火蟻攀爬率的影響
由表2可知,各處理組紅火蟻在相同處理時(shí)間下均以對(duì)照的攀爬率最高,且均隨處理時(shí)間的延長(zhǎng)紅火蟻攀爬率呈下降趨勢(shì),其中,曼陀羅干花和干葉對(duì)紅火蟻的影響最大,處理11 d后,曼陀羅干花和干葉處理小型紅火蟻的攀爬率分別為0和1.33%,其他部位提取物處理小型紅火蟻的攀爬率為21.33%~32.00%,與對(duì)照(65.33%)相比均達(dá)顯著差異水平;曼陀羅干花和干葉提取物處理中型紅火蟻的攀爬率均為0,其他部位提取物處理中型紅火蟻的攀爬率為8.00%~56.00%,與對(duì)照(89.33%)相比均達(dá)顯著差異水平。
2. 3 曼陀羅對(duì)紅火蟻?zhàn)ジ铰实挠绊?/p>
由表3可知,各處理組紅火蟻在相同處理時(shí)間下均以對(duì)照的抓附率最高,且均隨處理時(shí)間的延長(zhǎng)紅火蟻?zhàn)ジ铰手饾u下降,其中,曼陀羅干花和干葉對(duì)紅火蟻的影響最大,處理11 d后,曼陀羅干花和干葉處理小型紅火蟻的抓附率分別為0和1.33%,其他部位提取物處理小型紅火蟻的抓附率為18.67%~29.33%,與對(duì)照(61.33%)相比均達(dá)顯著差異水平;曼陀羅干花和干葉提取物處理中型紅火蟻的抓附率均為0,其他部位提取物處理中型紅火蟻的抓附率為6.67%~54.67%,與對(duì)照(88.00%)相比均達(dá)顯著差異水平。
2. 4 曼陀羅對(duì)紅火蟻聚集率的影響
由表4可知,各處理組紅火蟻在相同處理時(shí)間下均以對(duì)照的聚集率最高,且均隨處理時(shí)間的延長(zhǎng)紅火蟻聚集率呈下降趨勢(shì),其中,曼陀羅干花和干葉對(duì)小型紅火蟻的影響最大,處理7 d后小型紅火蟻的聚集率即降為0,其他部位提取物處理11 d后小型紅火蟻的聚集率為14.67%~28.00%,且與對(duì)照(60.00%)間存在顯著差異;曼陀羅干花、干葉和鮮葉提取物處理對(duì)中型紅火蟻的影響最大,分別在處理7、11和11 d后紅火蟻的聚集率降為0,其他部位提取物處理11 d后中型紅火蟻的聚集率為17.33%~34.67%,且與對(duì)照(69.33%)間存在顯著差異。
2. 5 曼陀羅對(duì)紅火蟻行走率的影響
由表5可知,各處理組紅火蟻在相同處理時(shí)間下均以對(duì)照的行走率最高,且均隨處理時(shí)間的延長(zhǎng)紅火蟻行走率逐漸下降,其中,曼陀羅干花對(duì)小型紅火蟻的影響最大,處理9 d后小型紅火蟻的行走率即降為0,其他部位提取物處理11 d后小型紅火蟻的行走率為1.33%~30.67%,且與對(duì)照(66.67%)相比達(dá)顯著差異水平;對(duì)中型紅火蟻行走率影響最大的是曼陀羅干花和干葉,分別于處理7和11 d后紅火蟻的行走率降為0,其他部位提取物處理11 d后中型紅火蟻的行走率為8.00%~58.67%,且與對(duì)照(90.67%)相比達(dá)顯著差異水平。
3 討論
曼陀羅是一種藥用植物,全株有毒,已廣泛應(yīng)用于醫(yī)學(xué)、農(nóng)業(yè)和林業(yè)領(lǐng)域(Alberto et al.,2001)。印度當(dāng)?shù)氐尼t(yī)生常用其提取物治療各種炎癥性疾?。∟avaratnarajah et al.,2007);其對(duì)粘蟲(chóng)和蚜蟲(chóng)等多種農(nóng)業(yè)害蟲(chóng)具有毒殺作用(李國(guó)平和楊鷺生,2002;孫洋等,2008)。在曼陀羅葉乙醇提取物作用下,菜青蟲(chóng)胃毒的死亡率可達(dá)92%,觸殺死亡率為84.4%,拒食率為62.5%,表現(xiàn)出良好的殺蟲(chóng)作用(高紅明等,1999)。曼陀羅種子能引起大鼠病理形態(tài)學(xué)改變(Verma et al.,2012),其東莨菪堿等對(duì)人體有極高的致幻作用。因此,以曼陀羅為植物材料來(lái)源,有望研發(fā)出有效的殺紅火蟻藥劑,但目前曼陀羅對(duì)紅火蟻行為和毒力的研究較少見(jiàn)報(bào)道。
本研究結(jié)果表明,曼陀羅不同部位的乙醇提取物對(duì)紅火蟻均有一定毒力,其中干花對(duì)兩種紅火蟻工蟻的毒殺效果最好,小、中型紅火蟻分別在處理11和7 d后死亡率達(dá)100.00%;其次是干葉,處理11 d后小、中型紅火蟻的死亡率分別為98.67%和100.00%,表現(xiàn)出較強(qiáng)的毒殺活性。曼陀羅不同部位提取物對(duì)紅火蟻的抓附率、攀爬率、聚集率和行走率也有抑制作用,會(huì)影響整個(gè)蟻群的繁衍及活動(dòng),與劉娜等(2011)的研究報(bào)道一致。本研究顯示,在曼陀羅干花作用下,處理11 d后兩種蟻型的攀爬率、抓附率、聚集率和行走率均為0;在曼陀羅干葉作用下,處理11 d后小型紅火蟻的攀爬率、抓附率和行走率均為1.33%,聚集率為0,而中型紅火蟻的攀爬率、抓附率、行走率和聚集率均為0,干花和干葉對(duì)紅火蟻均表現(xiàn)出良好的行為影響作用。本研究采用的乙醇提取法操作簡(jiǎn)單,且曼陀羅干葉和干花來(lái)源方便,具有一定的實(shí)際應(yīng)用意義。
本研究?jī)H探討了曼陀羅不同部位提取物在室內(nèi)條件下對(duì)小、中型紅火蟻工蟻的毒力和行為作用,但在田間的效果如何,是否可采用其他提取殺蟲(chóng)物質(zhì)的方式或拌餌料法、噴霧法等作用方式,是否可與其他植物源藥劑混用增強(qiáng)作用效果均有待進(jìn)一步研究證實(shí)。
4 結(jié)論
經(jīng)曼陀羅各部位提取物作用后兩種蟻型紅火蟻的行為能力均有所降低,對(duì)減輕紅火蟻的危害及對(duì)整個(gè)蟻群的防治均具有一定積極作用。曼陀羅各部位提取物中,干花對(duì)兩種蟻型紅火蟻的影響最明顯,具有開(kāi)發(fā)成新型綠色環(huán)保紅火蟻防治植物材料的潛力。
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(責(zé)任編輯 麻小燕)