三種生物源農(nóng)藥對桃樹蚜蟲的防治效果研究

喬 巖, 董 杰, 王品舒, 岳 瑾, 張保常, 張金良, 袁志強, 楊建國*

1.北京市植物保護站, 北京 100029;2.北京市平谷區(qū)植保站, 北京 101205

三種生物源農(nóng)藥對桃樹蚜蟲的防治效果研究

喬 巖1, 董 杰1, 王品舒1, 岳 瑾1, 張保常2, 張金良1, 袁志強1, 楊建國1*

1.北京市植物保護站, 北京 100029;2.北京市平谷區(qū)植保站, 北京 101205

為篩選生產(chǎn)上防治桃樹蚜蟲的生物源農(nóng)藥，開展了1.5%除蟲菊素水劑、0.3%苦參堿水劑和5%桉油精可溶液劑對桃樹蚜蟲的田間防效試驗。結(jié)果表明，3種生物源農(nóng)藥中以0.3%苦參堿水劑防治效果最好，藥后1 d防效為70.12%，藥后7 d防效為91.96%，與對照藥劑70% 吡蟲啉水分散粒劑防效相當，具有較好的速效性和持效性?？鄥A對桃樹安全，是防治桃蚜的理想藥劑，生產(chǎn)上推薦使用濃度為1 000倍液。

生物源農(nóng)藥；桃蚜；防效

桃樹蚜蟲主要有桃蚜(Myzuspersicae) 、桃粉蚜(Hylalopterusamygdali) 和桃瘤蚜(Myzusmomonis)，是危害桃、杏、李的3種主要害蟲，?；旌习l(fā)生危害。桃蚜被害葉皺縮卷曲，嚴重影響枝、葉的發(fā)育，其分泌物易生霉菌，傳播病毒；桃粉蚜危害葉片、新梢和果實，污染果面，引起葉、梢干枯，削弱樹勢; 桃瘤蚜可使葉面縱卷，腫脹扭曲，葉肉增厚，最后干枯脫落。3種蚜蟲世代多、繁殖量大、蔓延迅速、防治困難，是影響桃高產(chǎn)、穩(wěn)產(chǎn)的重要限制因素，尤其是樹齡大、管理粗放的桃園受害更重[1]。

常見防治桃樹蚜蟲的方法主要采取吡蟲啉、抗蚜威、蚜虱凈等農(nóng)藥進行化學防治，但污染環(huán)境，易導致農(nóng)藥殘留，影響果實質(zhì)量安全。由于長期使用化學藥劑防治桃蚜，導致桃蚜對多種藥劑產(chǎn)生了抗性，常用藥劑的防效明顯下降，防治變得越來越困難[2,3]。

生物源農(nóng)藥是從生物組織中提取的生物活性物質(zhì), 如苦參素、苦楝素和印楝素等, 其優(yōu)勢在于沒有化學農(nóng)藥所表現(xiàn)的諸多副作用。尋找安全、高效、殘效期長的生物源農(nóng)藥替代化學農(nóng)藥，已成為綠色、有機大桃生產(chǎn)急需破解的生產(chǎn)難題[4～5]。

為此，我們開展了3種生物源農(nóng)藥對桃蚜的田間藥效試驗，以期取得理想效果，為大桃生產(chǎn)中存在的蚜蟲問題，提供一種安全、有效的防治手段。

1 材料和方法

1.1 供試材料

供試藥劑為1.5%除蟲菊素水乳劑，由云南南寶植化有限責任公司生產(chǎn)，試驗藥液噴灑濃度為600倍液；0.3%苦參堿水劑由北京中捷四方生物科技有限公司生產(chǎn)， 試驗藥液噴灑濃度為1 000倍液；5%桉油精可溶液劑購自北京亞戈農(nóng)生物藥業(yè)有限公司，試驗藥液噴灑濃度為1 000倍液；對照藥劑70%吡蟲啉水分散粒劑購自陜西上格之路生物科學有限公司，試驗藥液噴灑濃度為2 000倍液。

1.2 試驗方法

試驗地選擇在平谷區(qū)夏各莊鎮(zhèn)陳太務村桃園中進行，樹齡12年。果園地勢平坦，光照條件好，長勢均勻，肥水管理水平一致。桃樹株行距4 m×5 m。試驗設5個處理，分別為1.5%除蟲菊素水乳劑600倍液，0.3%苦參堿水劑1 000倍液，5%桉油精可溶液劑1 000倍液，70%吡蟲啉水分散粒劑2 000倍液，空白對照噴清水。施藥時間為2014年4月3日，使用噴桿式噴霧器，噴霧均勻、徹底，達到整株淋洗狀態(tài)。2株樹為1個小區(qū)，重復3次，隨機排列。

1.3 調(diào)查時間和方法

噴藥前和噴藥后第1 d、3 d和7 d調(diào)查，共調(diào)查4次。調(diào)查小區(qū)內(nèi)2株樹，每株在東南西北中5個方位各標記一個有蟲嫩梢，每梢自頂向下調(diào)查5片葉(包括枝條)的活蟲數(shù)。藥效計算方法按下式計算，對照小區(qū)也按此法調(diào)查。

蟲口減退率(%) =[(藥前蟲口密度-藥后蟲口密度)÷藥前蟲口密度]×100%

防治效果(%)=[(處理蟲口減退率-對照蟲口減退率)÷(1-對照蟲口減退率)]×100%。

2 結(jié)果與分析

2.1 對桃蚜的防治效果

試驗結(jié)果表明(表1)，施藥后1 d，0.3%苦參堿水劑防治效果即達到70.12%,與對照藥劑70%吡蟲啉水分散粒劑防效相當，表現(xiàn)出較好的速效性。1.5%除蟲菊素水劑和5%桉油精可溶液劑防效分別為26.45%和19.63%，顯著低于對照藥劑70%吡蟲啉水分散粒劑2 000倍液。施藥后4 d，0.3%苦參堿水劑1 000倍液防治效果為69.7%，與藥后1 d相比，防效略有下降，與對照藥劑70%吡蟲啉水分散粒劑防效相當。1.5%除蟲菊素水劑和5%桉油精可溶液劑防效分別為31.28%、36.78%，與施藥后1 d相比，防效均顯著提高，仍顯著低于對照藥劑70%吡蟲啉水分散粒劑2 000倍液。藥后7 d，各藥劑處理防治效果均繼續(xù)增加，0.3%苦參堿水劑防治效果達到91.96%,與對照藥劑70%吡蟲啉水分散粒劑防效相當。1.5%除蟲菊素水劑和5%桉油精可溶液劑防效分別為65.89%和71.37%，顯著低于對照藥劑70%吡蟲啉水分散粒劑2 000倍液。不同藥劑對桃蚜的防治效果差異顯著。各藥劑隨施藥后隨時間增加防治效果提高，藥后7 d達到最好防治效果。

表1 4種藥劑防治桃樹蚜蟲效果

注:同列數(shù)據(jù)后不同小寫字母代表差異顯著(P<0.05)。

2.2 安全性

通過目測各處理藥劑對作物的安全性，施藥后1～7 d均未發(fā)現(xiàn)可觀察到的明顯藥害癥狀，果實未見異常或畸形果，可見3 種生物藥劑防治桃蚜未對果樹及果實產(chǎn)生明顯的不良影響，說明以上藥劑對大桃作物安全。

3 討論

田間試驗結(jié)果表明, 0.3%苦參堿水劑1 000倍液防治蚜蟲的效果非常理想, 藥后第7 d, 防效可達91.96%, 說明0.3%苦參堿水劑持效期長, 是防治蚜蟲的理想藥劑。從經(jīng)濟有效方面綜合考慮, 0.3%苦參堿水劑防治蚜蟲的田間使用劑量以1 000倍液為宜。

1.5%除蟲菊素水劑和5%桉油精可溶液劑藥后7 d防效分別達到65.89%和71.37%，與對照藥劑吡蟲啉相比防效差異明顯，對桃蚜的防治效果不甚理想。有研究表明，1.5%除蟲菊素水乳劑用于蔬菜蚜蟲的防治時，接觸到藥劑的部位效果較好，可能是由于該藥劑內(nèi)吸作用和滲透作用較差[6]。因此，桃園使用除蟲菊素水乳劑時，應注意均勻噴霧。5%桉油精可溶液劑目前用于防治林業(yè)害蟲較多，對槐蚜的防治效果一般[7]，對桃蚜的防治效果及使用技術(shù)還有待進一步研究。

2011年，宮亞軍等[8]研究發(fā)現(xiàn)，在北京地區(qū)，蔬菜上桃蚜對吡蟲啉仍具有很高的敏感性，北京地區(qū)普遍使用吡蟲啉做為防治桃蚜的首選藥物。本研究中，桃樹上的蚜蟲對吡蟲啉比較敏感，但是隨著使用時間的延長和用藥次數(shù)的增加，抗性也會發(fā)生?？梢圆扇≥啌Q用藥或者將不同作用機制的農(nóng)藥科學合理地復配混用，增加藥效，減少用藥頻次，省時省工，降低成本，延緩抗藥性產(chǎn)生[9]。根據(jù)本試驗的結(jié)果，苦參堿可以作為吡蟲啉的替代藥劑。

0.3%苦參堿水劑高效低毒, 持效期長, 防治蚜蟲效果好, 具有對人畜、環(huán)境、天敵等安全和果品無農(nóng)藥殘留、無三致(致癌、致畸、致突變)、無藥害、不易產(chǎn)生抗性等優(yōu)點, 符合綠色果品生產(chǎn)的要求[10]。因此, 建議在今后生產(chǎn)中大力推廣應用0.3%苦參堿水劑防治蚜蟲。

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[2] 陶 晡，康占海，李 星. 25%噻蟲嗪水分散粒劑防治桃蚜試驗[J]. 山西果樹，2014, 4：53-55.

[3] 李定旭. 兩種新型藥劑防治蘋果及桃樹蚜蟲藥效試驗[J]. 中國果樹，2002, 11：33-35.

[4] 蘇明申, 葉正文, 李勝源，等. 生物有機農(nóng)藥毒殺對桃樹蚜蟲的試驗[J]. 中國果樹，2008, 3：74.

[5] 杜肇南， 劉長祿. 一種新型植物源殺蟲劑的研制及桃樹蚜蟲防治試驗[J]. 中國果菜，2012, 10：20-21.

[6] 劉艷芝，馬井玉，徐祥文，等. 不同濃度天然除蟲菊素水乳劑防治蔬菜蚜蟲田間藥效試驗[J]. 長江蔬菜，2012, 12：70-71.

[7] 常承秀，朱惠英，張永強，等.8種藥劑防治槐蚜藥效試驗[J]. 森林保護，2013, 7：36-38.

[8] 宮亞軍，王澤華，石寶才, 等. 北京地區(qū)不同桃蚜種群的抗藥性研究[J]. 植物保護，2011, 44(21)：43-178.

[9] 劉雨晴，范 毅，于立芹, 等. 天然苦皮藤素和天然除蟲菊素混配對三種蚜蟲的毒力及田間防效[J]. 植物保護，2014, 40(2)：175-178.

[10] 竇立峰，王建玉，柳富華. 0.3%復方苦參堿水劑防治蚜蟲藥效試驗[J]. 山西果樹，2009, 129：11-12.

·Nature系列期刊導讀·

科學家通過增加靶點范圍來擴大CRISPR-Cas9技術(shù)的可用性

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論文鏈接: Benjamin P K,etal.. Broadening the targeting range ofStaphylococcusaureusCRISPR-Cas9 by modifying PAM recognition. Nature Biotechnology,2015,doi:10.1038/nbt.3404.

Abstract: CRISPR-Cas9 nucleases target specific DNA sequences using a guide RNA but also require recognition of a protospacer adjacent motif (PAM) by the Cas9 protein. Although longer PAMs can potentially improve the specificity of genome editing, they limit the range of sequences that Cas9 orthologs can target. One potential strategy to relieve this restriction is to relax the PAM recognition specificity of Cas9. Here we used molecular evolution to modify the NNGRRT PAM ofStaphylococcusaureusCas9 (SaCas9). One variant we identified, referred to as KKH SaCas9, showed robust genome editing activities at endogenous human target sites with NNNRRT PAMs, thereby increasing SaCas9 targeting range by two - to fourfold. Using GUIDE-seq, we show that wild-type and KKH SaCas9 induce comparable numbers of off-target effects in human cells. Our strategy for evolving PAM specificity does not require structural information and therefore should be applicable to a wide range of Cas9 orthologs.

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論文鏈接: Zhou X,etal.. Phage-mediated counting by the naked eye of miRNA molecules at attomolar concentrations in a Petri dish. Nature Materials, 2015, doi:10.1038/nmat4377.

Abstract: The ability to count biomolecules such as cancer-biomarker miRNAs with the naked eye is seemingly impossible in molecular diagnostics. Here, we show an ultrasensitive naked-eye-counting strategy for quantifying miRNAs by employing T7 phage—a bacteria-specific virus nanoparticle—as a surrogate. The phage is genetically engineered to become fluorescent and capable of binding a miRNA-capturing gold nanoparticle (GNP) in a one-to-one manner. Target miRNAs crosslink the resultant phage-GNP couple and miRNA-capturing magnetic microparticles, forming a sandwich complex containing equimolar phage and miRNA. The phage is then released from the complex and developed into one macroscopic fluorescent plaque in a Petri dish by plating it in a host bacterial medium. Counting the plaques by the naked eye enables the quantification of miRNAs with detection limits of ～3 and ～5 aM for single-target and two-target miRNAs, respectively. This approach offers ultrasensitive and convenient quantification of disease biomarkers by the naked eye.

新技術(shù)以可讓肽類藥物更加持久

近日，來自太平洋大學的科學家通過研究開發(fā)了一種新方法，可以明顯延長機體肽類的生命，該研究或為后期開發(fā)新型肽類藥物來治療癌癥及其他疾病提供思路。蛋白肽類可以被工程化操作來治療一系列疾病，包括癌癥和糖尿病等，這些小型的氨基酸鏈具有非常大的潛力和選擇性，而且相比大分子要更加安全一些，但肽類較短的壽命迫使科學家們不得不使用大劑量且頻繁給予患者來增加作用效果。該文中研究者利用新技術(shù)可以使得肽類壽命延長，這就可以降低肽類藥物的使用頻率，從而幫助患者節(jié)約藥物成本且降低患者的依從性。該項研究對于改善人類健康又邁進了一步，文章中采用的方法具有巨大的應用潛力，其不光可以幫助治療疾病，還可以增強對患者機體的成像及診斷。

論文鏈接: Sravan C P,etal.. A biomimetic approach for enhancing the in vivo half-life of peptides. Nature Chemical Biology,2015,doi:10.1038/nchembio.1907.

Abstract: The tremendous therapeutic potential of peptides has not yet been realized, mainly owing to their short in vivo half-life. Although conjugation to macromolecules has been a mainstay approach for enhancing protein half-life, the steric hindrance of macromolecules often harms the binding of peptides to target receptors, compromising the in vivo efficacy. Here we report a new strategy for enhancing the in vivo half-life of peptides without compromising their potency. Our approach involves endowing peptides with a small molecule that binds reversibly to the serum protein transthyretin. Although there are a few molecules that bind albumin reversibly, we are unaware of designed small molecules that reversibly bind other serum proteins and are used for half-life extension in vivo. We show here that our strategy was effective in enhancing the half-life of an agonist for GnRH receptor while maintaining its binding affinity, which was translated into superior in vivo efficacy.

微生物合作的新機制被揭示

海洋沉積物中甲烷的厭氧氧化，是由甲烷氧化古菌和能產(chǎn)生硫酸鹽的細菌共同完成的一個協(xié)作過程。人們曾認為，一種可溶性代謝物在合作的微生物之間的交換是必不可少的，但在Victoria及同事最近的研究結(jié)果否定了這一假設。他們在單細胞層面上，對來自西北太平洋中一個活動甲烷滲出泉取樣的沉積物制備的微生物混合體中的生物合成活性做了相關研究。他們發(fā)現(xiàn)，細胞活動是獨立于互養(yǎng)伙伴之間的距離的，這與涉及中間體短距離擴散的一個模型是不一致的。實際上，古菌與細菌之間的直接電子轉(zhuǎn)移，才是它們相互作用的核心機制。

論文鏈接: Mcglynn S E,etal.. Single cell activity reveals direct electron transfer in methanotrophic consortia. Nature, 2015,doi: 10.1038/nature15512.

Abstract: Multicellular assemblages of microorganisms are ubiquitous in nature, and the proximity afforded by aggregation is thought to permit intercellular metabolic coupling that can accommodate otherwise unfavourable reactions. Consortia of methane-oxidizing archaea and sulphate-reducing bacteria are a well-known environmental example of microbial co-aggregation; however, the coupling mechanisms between these paired organisms is not well understood, despite the attention given them because of the global significance of anaerobic methane oxidation. Here we examined the influence of interspecies spatial positioning as it relates to biosynthetic activity within structurally diverse uncultured methane-oxidizing consortia by measuring stable isotope incorporation for individual archaeal and bacterial cells to constrain their potential metabolic interactions. In contrast to conventional models of syntrophy based on the passage of molecular intermediates, cellular activities were found to be independent of both species intermixing and distance between syntrophic partners within consortia. A generalized model of electric conductivity between co-associated archaea and bacteria best fit the empirical data. Combined with the detection of large multi-haem cytochromes in the genomes of methanotrophic archaea and the demonstration of redox-dependent staining of the matrix between cells in consortia, these results provide evidence for syntrophic coupling through direct electron transfer.

生物多樣性喪失威脅生態(tài)系統(tǒng)穩(wěn)定性

近日，研究者們將來自46個實驗的數(shù)據(jù)相結(jié)合發(fā)現(xiàn)，生物多樣性會提高一個生態(tài)系統(tǒng)對極端氣候事件的抵抗力。只有少量物種的地塊，在極端氣候事件期間生產(chǎn)力下降了50%，而這種影響隨著物種數(shù)量的增加會被減半。然而，生物多樣性對生態(tài)系統(tǒng)彈性沒有明顯影響，低生物多樣處理和高生物多樣性處理，在一年內(nèi)，都能夠從極端氣候事件中恢復過來。

論文鏈接: Nico E,etal.. Biodiversity increases the resistance of ecosystem productivity to climate extremes. Nature, 2015, doi: 10.1038/nature15374.

Abstract: It remains unclear whether biodiversity buffers ecosystems against climate extremes, which are becoming increasingly frequent worldwide. Early results suggested that the ecosystem productivity of diverse grassland plant communities was more resistant, changing less during drought, and more resilient, recovering more quickly after drought, than that of depauperate communities. However, subsequent experimental tests produced mixed results. Here we use data from 46 experiments that manipulated grassland plant diversity to test whether biodiversity provides resistance during and resilience after climate events. We show that biodiversity increased ecosystem resistance for a broad range of climate events, including wet or dry, moderate or extreme, and brief or prolonged events. Across all studies and climate events, the productivity of low-diversity communities with one or two species changed by approximately 50% during climate events, wheras that of high-diversity communities with 16～32 species was more resistant, changing by only approximately 25%. By a year after each climate event, ecosystem productivity had often fully recovered, or overshot, normal levels of productivity in both high-and low-diversity communities, leading to no detectable dependence of ecosystem resilience on biodiversity. Our results suggest that biodiversity mainly stabilizes ecosystem productivity, and productivity-dependent ecosystem services, by increasing resistance to climate events. Anthropogenic environmental changes that drive biodiversity loss thus seem likely to decrease ecosystem stability, and restoration of biodiversity to increase it, mainly by changing the resistance of ecosystem productivity to climate events.

青蒿素類過氧橋鍵的生物合成機制被解析

青蒿素的生物活性與過氧鍵密不可分，但是催化青蒿酸形成青蒿素的環(huán)內(nèi)過氧鍵合酶卻一直沒有找到，成為一道世界難題。科學家們最近從幾株曲霉和青霉菌種中分離出具有抗感染等多種生物活性的萜類吲哚生物堿真菌毒素Verruculogen。文章首次報道了FtmOx1的晶體結(jié)構(gòu)，以及FtmOx1分別與a-酮戊二酸和底物fumitremorgen B的共晶體結(jié)構(gòu)，并通過詳盡的酶學實驗結(jié)果驗證了FtmOx1的功能?？焖俜磻獎恿W和冷凍淬火電子自旋光譜實驗結(jié)果證實了FtmOx1反應中存在自由基中間體。闡明這一特別的環(huán)內(nèi)過氧橋鍵的生物合成新機制為發(fā)現(xiàn)催化青蒿酸形成青蒿素的環(huán)內(nèi)過氧鍵合酶向前邁進了一大步。進一步研究其酶學機制，將為含有過氧橋鍵的萜類吲哚生物堿的廣泛應用奠定科學和應用基礎。

論文鏈接: Yan W P,etal.. Endoperoxide formation by an α-ketoglutarate-dependent mononuclear non-haem iron enzyme. Nature, 2015, doi:10.1038/nature15519.

Abstract: Many peroxy-containing secondary metabolites have been isolated and shown to provide beneficial effects to human health. Yet, the mechanisms of most endoperoxide biosyntheses are not well understood. Although endoperoxides have been suggested as key reaction intermediates in several cases, the only well-characterized endoperoxide biosynthetic enzyme is prostaglandin H synthase, a haem-containing enzyme. Fumitremorgin B endoperoxidase (FtmOx1) from Aspergillus fumigatus is the first reported α-ketoglutarate-dependent mononuclear non-haem iron enzyme that can catalyse an endoperoxide formation reaction. To elucidate the mechanistic details for this unique chemical transformation, we report the X-ray crystal structures of FtmOx1 and the binary complexes it forms with either the co-substrate (α-ketoglutarate) or the substrate (fumitremorgin B). Uniquely, after α-ketoglutarate has bound to the mononuclear iron centre in a bidentate fashion, the remaining open site for oxygen binding and activation is shielded from the substrate or the solvent by a tyrosine residue (Y224). Upon replacing Y224 with alanine or phenylalanine, the FtmOx1 catalysis diverts from endoperoxide formation to the more commonly observed hydroxylation. Subsequent characterizations by a combination of stopped-flow optical absorption spectroscopy and freeze-quench electron paramagnetic resonance spectroscopy support the presence of transient radical species in FtmOx1 catalysis. Our results help to unravel the novel mechanism for this endoperoxide formation reaction.

Control Effects of Three Biogenic Pesticides on Controlling Peach Aphids

QIAO Yan1, DONG Jie1, WANG Pin-shu1,YUE Jin1, ZHANG Bao-chang2, ZHANG Jin-liang2, YUAN Zhi-qiang1, YANG Jian-guo1*

1.BeijingPlantProtectionStation,Beijing100029,China;2.BeijingPingguDistrictPlantProtectionStation,Beijing101205,China

To screen biogenic pesticides in controlling aphids on peach tree, the field test was carried out using 1.5% pyrethrin aqueous solution (AS), 0.3% matrine AS and 5% eucalyptole soluble concentrate against aphids in the field. The results indicated that of the three biogenic pesticides, 0.3% matrine AS was the most effective. The control efficiency could reach 70.12% on the first day and 91.96% on the 7th day respectively after spraying. Comparing with 70% imidacloprid water dispersible granule, the control effects of 0.5% matrine AS were not significantly different. Matrine AS was well quick-acting and persistance on the control of aphids. The matrine AS was safe to peach tree, and was ideal pesticide to control aphids. The 1 000 dilution of 0.3% matrine AS was suggested to be applied in the fields.

biogenic pesticides; peach aphids; control efficiency

2015-09-08; 接受日期：2015-10-28

農(nóng)作物病蟲鼠害疫情監(jiān)測與防治項目資助。

喬巖，高級農(nóng)藝師，研究方向為糧經(jīng)作物病蟲害防治。E-mail：qy0914@126.com。*通信作者：楊建國，推廣研究員，主要從事糧食、經(jīng)濟作物病蟲害綠色防控技術(shù)研究及示范推廣工作。E-mail：yangjguo@263.net

10.3969/j.issn.2095-2341.2015.06.12