發(fā)布時間：2017-12-28 00:34

  本文關(guān)鍵詞：小貫小綠葉蟬的疏水性及其在控制中的潛在應(yīng)用　出處：《福建農(nóng)林大學(xué)》2017年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 小貫小綠葉蟬 噴霧刺激 疏水性 粘附力 無害化防治

【摘要】：小貫小綠葉蟬Empoasca onukii Matsuda是我國茶園的重要害蟲,目前已對多種農(nóng)藥品種產(chǎn)生抗藥性,既增加了防治難度,同時也因農(nóng)藥殘留等問題影響茶葉飲用的安全性,急需尋找無害化防治措施替代傳統(tǒng)的化學(xué)防治。小貫小綠葉蟬具有畏水習(xí)性,受降雨等水分脅迫影響較大,另一方面該葉蟬活動性強(qiáng),受驚擾時善于躲避,農(nóng)藥的噴霧防治效果常受到影響。本試驗(yàn)通過室內(nèi)人工模擬降雨分別對成若蟲進(jìn)行噴霧刺激,研究成蟲和若蟲的疏水性行為反應(yīng)特點(diǎn),分析成、若蟲不同部位的微觀結(jié)構(gòu),并探索噴霧防治該葉蟬可行性,主要研究結(jié)果如下:(1)觀察發(fā)現(xiàn)小貫小綠葉蟬成蟲和若蟲對清水噴霧刺激存在不同的行為反應(yīng)。試驗(yàn)首先比較齡期對若蟲逃避行為的影響,1齡若蟲會被噴霧到葉片的水流沖下,其他齡期若蟲則困于葉片水泊中;分析停止噴霧后若蟲逃離水滴的時間,發(fā)現(xiàn)高齡若蟲(4～5齡)的逃離時間17.40±2.78s,顯著短于低齡若蟲(2～3齡)48.70 ± 3.05 s,葉片正面形成的水滴比葉背水膜更有利于若蟲逃離;逃離視頻觀察發(fā)現(xiàn)若蟲腹部與水滴間存在較高的粘附力。其次,對成蟲群體進(jìn)行加罩噴霧刺激,成蟲可躲避在亞克力板上密集水滴的空隙之間,當(dāng)罩內(nèi)加上紗網(wǎng)并噴霧形成水膜后,80%成蟲不發(fā)生逃避行為而困于葉面水滴中,20%成蟲前后翅粘附在紗網(wǎng)水膜上,微距觀察發(fā)現(xiàn)成蟲的雙翅與水膜間也存在較高的粘附力。研究主要器官對成蟲逃避行為的影響,完好組成蟲可在0.29 ± 0.15 s內(nèi)快速逃離噴霧區(qū)域,復(fù)眼遮蓋組成功逃離的概率最低,67%試蟲困于葉片水膜中,觸角及雙翅對其逃避行為的影響較小。(2)證明小貫小綠葉蟬成蟲和若蟲前跗節(jié)結(jié)構(gòu)不同與其疏水性差異存在必然聯(lián)系。通過水面法觀察小貫小綠葉蟬成、若蟲前跗節(jié)的疏水性,并與茶二叉蚜和白背飛虱進(jìn)行對比,結(jié)果顯示葉蟬成蟲可站立于水面,而若蟲前跗節(jié)則陷入水中,表明成蟲前跗節(jié)的疏水性高于若蟲,相比之下,茶二叉蚜和白背飛虱不同蟲態(tài)間的疏水性沒有差異,茶二叉蚜成、若蟲的前附節(jié)均陷入水中,白背飛虱成、若蟲則均可站立于水面。通過掃描電鏡比較分析試蟲前跗節(jié)微觀結(jié)構(gòu)的差異,小貫小綠葉蟬成蟲前跗節(jié)較為寬扁,爪中墊為兩個淺盤結(jié)構(gòu),上面覆蓋大量網(wǎng)粒體,而若蟲前跗節(jié)較肥厚,寬度和厚度與成蟲均存在顯著差異,爪中墊為兩個中空吸杯,而茶二叉蚜和白背飛虱成、若蟲兩種蟲態(tài)的前跗節(jié)結(jié)構(gòu)沒有顯著差異。通過自制簡易吸管天平測定小貫小綠葉蟬前跗節(jié)與茶葉葉面及水滴的粘附力,結(jié)果顯示若蟲前跗節(jié)與茶葉葉面的粘附力達(dá)96.7 ± 6.9 μN(yùn),與水面的粘附力達(dá)108.6±11.9 μN(yùn),都顯著高于成蟲。我們推測若蟲前跗節(jié)吸杯式爪中墊為若蟲與接觸界面提供較高的吸附力,使其在噴霧過程中不會被水流沖下,而成蟲前跗節(jié)的寬扁結(jié)構(gòu)及存在的大量網(wǎng)粒體提高其疏水性,有利于成蟲快速逃離噴霧區(qū)域。(3)證實(shí)小貫小綠葉蟬腹部表皮的顯微結(jié)構(gòu)隨發(fā)育而變化,覆蓋的網(wǎng)粒體數(shù)量也隨之增多,疏水性逐漸增強(qiáng)。試驗(yàn)觀察到若蟲腹部背面除了 6列感覺毛外,表皮較為光滑,腹面微觀結(jié)構(gòu)則隨蟲齡而變化,低齡若蟲(1～2齡)腹部腹面密布著大量的微乳突(0.3～0.8 μm),中高齡若蟲(3～5齡)腹部中央的微乳突轉(zhuǎn)變?yōu)槲⒋?0.7～1.6 μm);成蟲腹部腹面和背面均有魚鱗狀排列的微刺,腹面微刺長度(0.7～1.6 μm)比背板微刺(0.3～0.6 μm)稍長。成、若蟲腹部不同的微觀結(jié)構(gòu)與網(wǎng)粒體的覆蓋密度有很大關(guān)系。SEM圖像顯示成蟲腹部的網(wǎng)粒體數(shù)量顯著多于若蟲,且分布更為均勻。通過測定葉蟬腹部的疏水性,發(fā)現(xiàn)網(wǎng)粒體移除前成蟲腹部靜態(tài)接觸角CAs值平均值為149.10 ±2.36°,去除網(wǎng)粒體后CAs平均值為125.35 ±3.04°,網(wǎng)粒體移除對成蟲腹部的疏水性影響較大,而對若蟲影響較小。通過比較小貫小綠葉蟬成蟲和若蟲腹部與水的粘附力POF值,若蟲腹部腹面的POF值最高,網(wǎng)粒體可顯著降低蟲體表皮與水的粘附力。通過以上結(jié)果,我們推測若蟲腹部與水的粘附力產(chǎn)生原因與蟲體太小及網(wǎng)粒體分布不均勻有關(guān),細(xì)小水滴在若蟲體表能夠形成近球形的狀態(tài),表現(xiàn)出較強(qiáng)的疏水性,符合Cassie模型,而較大水滴則浸潤到若蟲腹部其他部位,呈明顯的Wenzel狀態(tài),表現(xiàn)出較強(qiáng)的粘附力。(4)揭示小貫小綠葉蟬翅面覆蓋的網(wǎng)粒體具有提高翅面疏水性以及導(dǎo)致翅面與水間形成高粘附力的雙重作用。小貫小綠葉蟬前后翅經(jīng)掃描電鏡觀察顯示都均勻覆蓋網(wǎng)粒體。前后翅與水的靜態(tài)接觸角經(jīng)測定CAs150°,表明都具有超疏水性;去除網(wǎng)粒體后,前后翅翅面的疏水性水平顯著降低。小貫小綠葉蟬成蟲前后翅與水的粘附力達(dá)160.3 ± 3.5 μN(yùn),接近其自身重力的33倍。這種具超疏水性同時又具有高粘附力的現(xiàn)象屬于"花瓣效應(yīng)",形成原因與覆蓋在葉蟬翅面微米級的網(wǎng)粒體以及網(wǎng)粒體表面納米級的空隙所構(gòu)成微納米二級結(jié)構(gòu)有關(guān),與水接觸時網(wǎng)粒體內(nèi)部所截留的空氣形成翅面與水間的強(qiáng)粘附力。(5)根據(jù)小貫小綠葉蟬若蟲和成蟲不同的疏水性特點(diǎn),探索獲得了清水噴霧防治若蟲的最佳措施,以及加罩噴霧防治成蟲對減少殺蟲劑用量的具體效果。首先測定小貫小綠葉蟬不同發(fā)育階段對浸泡水滴的耐受力,發(fā)現(xiàn)低齡若蟲(2～3齡)的耐受力最弱,48h死亡率達(dá)到83.3%,而高齡若蟲和成蟲對浸泡水滴的耐受力都較強(qiáng),死亡率較低,因此推薦生產(chǎn)上采用噴霧法防控小貫小綠葉蟬若蟲。室內(nèi)試驗(yàn)初步獲得當(dāng)噴霧時間為20 min,噴霧距離為30～40 cm,沖刷次數(shù)為2次,72h后可獲得若蟲較高的致死率75%。其次,在室內(nèi)采用液滴法和紗網(wǎng)藥膜法測定成蟲雙翅被困于藥液的毒力效果,并用浸葉法對比藥劑的胃毒毒力,用離心管藥膜法對比藥劑的觸殺毒力,所測3種藥劑在濃度較低時即獲得較高的致死率,其中印楝素的紗網(wǎng)藥膜法LC50值為43.39μg/L,為制劑稀釋6.9 × 104倍液的濃度,蘇云金桿菌的LC50值為447.2IU/L,濃度為制劑稀釋1.7×107倍,標(biāo)準(zhǔn)藥劑聯(lián)苯菊酯的LC50值為0.12 μg/L,為制劑稀釋2.0 ×108倍,表明所測藥劑采用紗網(wǎng)藥膜法和液滴法均具有較強(qiáng)的胃毒及觸殺效果。此外,設(shè)計(jì)可移動式薄膜紗網(wǎng)噴霧罩開展田間試驗(yàn),添加低濃度印楝素乳油(稀釋1500倍)的噴霧防控效果接近50%,但與不加罩的噴霧防效差異不顯著,因此加罩減藥的實(shí)際效益還需篩選其他觸殺性更強(qiáng)的藥劑進(jìn)行驗(yàn)證。目前關(guān)于昆蟲的疏水性研究多應(yīng)用于仿生學(xué)材料,如何利用昆蟲的疏水性特點(diǎn)為害蟲的農(nóng)業(yè)防治提供有力支撐所見甚少,本試驗(yàn)研究結(jié)果將為農(nóng)業(yè)小型害蟲的無害化防治提供新的思路。
[Abstract]:Through the small green leafhopper Empoasca onukii Matsuda is an important pest of tea in China, has a variety of pesticide resistance, which increased the difficulty of safety control, but also because of the problem of pesticide residues of tea drinking, the urgent need to find harmless prevention measures instead of the traditional chemical control. The small green leafhopper has the fear of water habits, and is greatly influenced by rainfall and other water stress. On the other hand, the leafhopper is strong in activity and is good at avoiding when disturbed. Pesticide spraying and control effect is often affected. This experiment through the indoor artificial rainfall simulation respectively on nymph spray stimulation, characteristics of hydrophobic behavioral response of adult and nymph, analysis and microstructure of different parts of the nymphs, and explore the feasibility of spray control leafhopper, the main results are as follows: (1) observed through small green leafhopper nymphs and adults on the water spray stimulation has some different behavioral responses. Effect of age on the first nymphal escape behavior, 1 instar nymphs will be sprayed onto the leaves washed down, other nymphs are trapped in the leaves in the spray is stopped after the analysis of water; water nymph escape time, found (4 to 5 instar nymphs of age) from 17.40 + 2.78s, was significantly shorter the nymph (2 ~ 3 years) 48.70 + 3.05 s, the formation of water droplets than leaf blade back water film is more conducive to escape video observation nymphs flee; adhesion exists between water droplets and higher abdominal nymphs. Secondly, the adult population is covered with spray stimulation, adults can escape in acrylic plate water intensive gap between, when the water film is formed in the cover with gauze and spray, 80% adults without escape behavior while trapped in leaf drop, 20% adult wing adhesion in the screen above the water film, macro observation showed that the adhesion force is higher there adults and the water film between the wings. The effects of main organs on adults' escaping behavior were studied. Intact insects could escape quickly from 0.29 to 0.15 s, and the probability of escape was 67%. (2) it is proved that the difference of the structure of the tarsus in the adult and nymph of the small green leafhopper is related to the difference of its hydrophobicity. The surface was observed through small green leafhopper nymphs, pretarsus hydrophobic, and compared with two tea graminum and WBPH, results show that can stand on the surface of adult leafhoppers, and nymphs pretarsus into the water, that the adult hydrophobic tarsi than nymphs, compared with there is no difference between the two, hydrophobic tea graminum and sogatellafurcifera different insect state between the two, tea graminum nymphs of the sections are immersed into water, sogatellafurcifera, nymphs could stand in the water. Through the comparative analysis of differences from scanning electron microscopy pretarsus micro structure, through the small green leafhopper adults pretarsus is wide and flat, two claw pad tray structure, covered a large number of network powders, and nymphs pretarsus than hypertrophy, width and thickness of adults and there are significant differences in jaw mat two hollow suction cup, there is no significant difference between the pretarsus structure and two fork tea aphid and WBPH nymphs, two stages. The adhesion of self-made small tubular Straw balance determination of leafhopper pretarsus and tea leaf surface and water droplets, results show that the adhesion tarsi and nymphs before tea leaves up to 96.7 + 6.9 N 108.6 + and the surface adhesion force of 11.9 N, were significantly higher than that of adults. We hypothesized that provide higher adsorption power nymph pretarsus cup type claw pad for nymph and contact interface, the spray process will not be washed, and improve the hydrophobicity of wide tarsal adult before the flat structure and the existence of a large network of particles, is conducive to adult quickly fled the spray area. (3) the microscopic structure of the abdominal epidermis of the leafhopper was confirmed to change with its development, and the number of the covered net particles increased and the hydrophobicity increased gradually. Experimental observation on the back of the abdomen to nymphs in addition to the 6 column sensory hair, the skin is smooth, ventral microstructure with larval age changes, nymph (1 ~ 2 years) with a large number of abdominal ventral micropapillae (0.3 ~ 0.8 m), middle age (3 to 5 instar nymphs) micro mastoid central abdomen into the micro (0.7 ~ 1.6 m); adult abdomen adaxially and abaxially were arranged slightly scaly, ventral micro length (0.7 ~ 1.6 m) than micro backplane (0.3 ~ 0.6 m) slightly longer. The microstructure of the adult and nymphs has a great relationship with the coverage density of the net particles. SEM images showed that the number of net particles in the abdomen of the adult was more than that of the nymphs, and the distribution was more uniform. Through the determination of hydrophobic leafhoppers in the abdomen, found the net particles removed before the adult abdominal static contact angle the average CAs value of 149.10 plus or minus 2.36 degrees, the removal of net particles after the average value of CAs is 125.35 + 3.04 degrees, affect the particle removal of the hydrophobic network adult abdomen, while smaller influence on the nymphs. By comparing the small green leafhopper nymphs and adults through abdominal adhesion and water POF value, the highest POF value of abdominal ventral nymphs, adhesion of net particles can significantly reduce the body skin with water. According to the above results, we speculate that the adhesion of abdomen and causes water nymphs and worms are too small and uneven distribution of particles, small droplets can form nearly spherical in nymph surface, showed strong hydrophobicity, with the Cassie model, while the larger droplets are infiltrating into other parts of the abdomen showed a nymph, obviously Wenzel, showed strong adhesion. (4) the net particles covered by the wing surface of the small green leafhopper can increase the hydrophobicity of the wing surface and lead to the high adhesion between the wing surface and the water. The scanning electron microscope (SEM) of the front and back wings of the leafhopper of the small intersecting green leafhopper showed that the net particles were evenly covered. The static contact angle between the front and back wings and water is measured CAs150 degrees, indicating that all of them have super hydrophobicity. After removing the net particles, the hydrophobicity of the wing wing surface of the front and back wings
【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：S435.711

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