本文選題：楚雄腮扁葉蜂 + 蟲生真菌��； 參考：《西南林業(yè)大學(xué)》2015年碩士論文

【摘要】：楚雄腮扁葉蜂(Cephalcia chuxiongica)是云南省一種新的針葉林食葉害蟲。由于該蟲危害重、擴(kuò)散快、防治難,已成為近年來(lái)嚴(yán)重制約云南林業(yè)可持續(xù)發(fā)展的危險(xiǎn)性害蟲之一。至今未見(jiàn)對(duì)其科學(xué)有效防治方法的報(bào)道。蟲生真菌是害蟲種群自然調(diào)節(jié)的重要因子和害蟲生物防治的重要材料,利用蟲生真菌防治害蟲,已逐步成為一種以生物防治為主導(dǎo)的綜合治理重要途徑。旨在為該害蟲的生物防治提供菌種資源,本文開展了其蟲生真菌的分離、鑒定及高致病性菌株的篩選研究,取得了如下主要研究結(jié)果:(1)蟲生真菌的分離與鑒定。采用組織分離法,從云南省尋甸縣采集到的自然罹病死亡的楚雄腮扁葉蜂幼蟲蟲尸上分離到5株蟲生真菌:SWYH01,SWYH02,SWYH06,SWYH07和SWYH09;經(jīng)形態(tài)學(xué)和分子生物學(xué)方法鑒定,5株菌分別為布氏白僵菌Beauveria brongniartii(Sacc.)Petch、淡紫擬青霉Paecilomyces lilacinus(Thom)Samson)、淡色叢赤殼Bionectria ochroleuca(Schwein)SchroersSamuels、粉質(zhì)棒束孢Isaria farinose(Holmsk.)Fr.)、球孢白僵菌Beauveria bassiana(Balsamo)Vuillemin。(2)高致病性菌株的篩選。以經(jīng)回接驗(yàn)證后對(duì)楚雄腮扁葉蜂幼蟲具有致病性的5株蟲生真菌為研究對(duì)象,分別以5株菌的濃度為108個(gè)孢子/mL的孢子懸浮液對(duì)楚雄腮扁葉蜂幼蟲進(jìn)行致病力測(cè)試。通過(guò)對(duì)5株菌的主要侵染部位觀察,發(fā)現(xiàn)除菌株SWYH06是主要從氣門侵入,其余4株菌首先主要侵染楚雄腮扁葉蜂幼蟲的體壁、節(jié)間褶以及頭和尾等部位。處理16d后,結(jié)果表明,該5株蟲生真菌對(duì)楚雄腮扁葉蜂幼蟲均有一定的致病效果,菌株SWYH06對(duì)楚雄腮扁葉蜂幼蟲的致病速率、校正死亡率及僵蟲率都高于其余4株菌;死亡率、校正死亡率都可達(dá)100%,僵蟲率可達(dá)91.65%。菌株SWYH02對(duì)楚雄腮扁葉蜂的致病力最弱,死亡率和校正死亡率都不到50%,僵蟲率為25%。(3)菌株SWYH06的生物學(xué)特性研究。通過(guò)對(duì)菌絲生長(zhǎng)和產(chǎn)孢的營(yíng)養(yǎng)及環(huán)境條件篩選,確定其菌絲生長(zhǎng)和產(chǎn)孢的最適培養(yǎng)基為PPDA;實(shí)驗(yàn)所選擇的碳源對(duì)菌絲生長(zhǎng)的影響差異不明顯,菌絲生長(zhǎng)的最適氮源、無(wú)機(jī)鹽分別是酵母粉和MnSO4,產(chǎn)孢最適碳源是葡萄糖、氮源是NH4NO3和尿素、無(wú)機(jī)鹽是ZnSO4和MgSO4;在實(shí)驗(yàn)所設(shè)置的溫度范圍內(nèi),菌絲生長(zhǎng)、產(chǎn)孢及孢子萌發(fā)的適宜溫度是25℃;分生孢子萌發(fā)的較適濕度為90%以上;光照對(duì)菌絲生長(zhǎng)、孢子萌發(fā)影響不明顯,但可促進(jìn)菌株產(chǎn)孢。(4)菌株SWYH06對(duì)楚雄腮扁葉蜂和黃粉蟲(Tenebrio molitor)的毒力測(cè)定。用SWYH06菌株不同濃度梯度的孢子懸浮液處理楚雄腮扁葉蜂幼蟲和黃粉蟲幼蟲,結(jié)果表明,2種幼蟲的死亡率隨孢子濃度的增加而增加,死亡率與孢子懸浮液的濃度呈正相關(guān)。采用1.5×108個(gè)孢子/mL的孢子懸浮液處理楚雄腮扁葉蜂和黃粉蟲幼后,幼蟲的死亡高峰分別出現(xiàn)在第4~6d和2~4d;死亡率和僵蟲率分別為98.35%、96.65%和93.33%、96.65%。(5)楚雄腮扁葉蜂的室內(nèi)防治實(shí)驗(yàn)。使用成分為棉籽殼:麥麩:鋸末:葡糖糖:CaSO4=40:40:18:1:1的固體培養(yǎng)基,對(duì)SWYH06菌株進(jìn)行固體發(fā)酵。結(jié)果表明,SWYH06菌株在該培養(yǎng)基上菌絲生長(zhǎng)和產(chǎn)孢情況良好,10~15d菌絲可長(zhǎng)滿瓶,第20d時(shí),固體發(fā)酵物的含孢量達(dá)到4.5×108個(gè)孢子/g~6.7×108個(gè)孢子/g。用3種不同施菌方法對(duì)楚雄腮扁葉蜂進(jìn)行了處理。結(jié)果表明,使用固體發(fā)酵物拌土和在幼蟲入土前在土表層施用固體發(fā)酵物2種方法處理楚雄腮扁葉峰幼蟲,3個(gè)月后,校正死亡率可達(dá)74.08±7.64%和63.33±12.58%,說(shuō)明SWYH06菌株在楚雄腮扁葉蜂生物防治方面具有良好的應(yīng)用前景。
[Abstract]:The Cephalcia chuxiongica (Cephalcia chuxiongica) is a new leaf eating pest of coniferous forest in Yunnan province. Because of its heavy harm, rapid spread and difficult control, it has become one of the dangerous pests which seriously restrict the sustainable development of forestry in Yunnan in recent years. The important factors of the festival and the important material of pest biological control, and the use of entomophyll fungi to prevent and control pests have gradually become an important way to control the biological control. The aim is to provide bacterial resources for the biological control of the pest. The isolation, identification and screening of highly pathogenic strains of the insect fungi are carried out in this paper. The main results are as follows: (1) isolation and identification of entomotic fungi. Using tissue separation method, 5 strains of entomophytic fungi were isolated from the naturally sick and dead SWYH01 gills larvae from Xundian County of Yunnan Province: SWYH01, SWYH02, SWYH06, SWYH07 and SWYH09; 5 strains of bacteria were identified by morphological and molecular biological methods. Beauveria brongniartii (Sacc.) Petch, Paecilomyces lilicus Paecilomyces lilacinus (Thom) Samson), light colored plexus Bionectria ochroleuca (Schwein) SchroersSamuels, powdery Beauveria bassiana (2) screening of highly pathogenic strains. The pathogenicity of 5 strains of entomotic fungi in the larva of Chuxiong gills were tested. The pathogenicity test was carried out on the larvae of Chuxiong gills by 5 strains of spore suspension with 108 spores /mL, respectively. Through the observation of the main infection sites of 5 strains of bacteria, it was found that the other 4 strains were mainly from the valve and the other 4 strains. First, it mainly infects the body wall, internode fold and head and tail parts of the larva of Chuxiong parietal beeleaf bee. After treating 16d, the results show that the 5 strains of entomotic fungi have certain pathogenic effect on the larva of Chuxiong parietal beeleaf bee, and the strain SWYH06 is higher than the rest of the other 4 strains, and the death rate and the rate of ossified insect are higher than those of the other 4 strains. The rate of correction can reach 100%, and the rate of 91.65%. strain SWYH02 is the weakest, the mortality and correction mortality are less than 50%, and the biological characteristics of 25%. (3) strain SWYH06 are studied. The growth and sporulation of mycelia and the sporulation are selected to determine the growth and sporulation of the mycelia. The optimum medium is PPDA; the difference of the influence of the carbon source selected by the experiment is not obvious. The most suitable nitrogen source of mycelial growth, the inorganic salt is yeast powder and MnSO4, the most suitable carbon source of the sporulation is glucose, the nitrogen source is NH4NO3 and urea, the inorganic salt is ZnSO4 and MgSO4; in the temperature range set, the mycelium growth, spore producing and spores The suitable temperature for germination was 25, and the suitable humidity of conidia germination was more than 90%. The effect of light on mycelium growth and spores germination was not obvious, but it could promote the sporulation. (4) the virulence of strain SWYH06 to Chuxiong cheek flattened bee and Huang Fenchong (Tenebrio molitor). The treatment of Chuxiong gills with SWYH06 strains with different concentration gradient of spore suspension. The results showed that the mortality of the 2 larvae increased with the increase of spores concentration, and the mortality rate was positively correlated with the concentration of spore suspension. The death peaks of the larvae were in 4~6d and 2~4d respectively after the treatment of 1.5 x 108 spores /mL of spore suspension. And the rate of ossified insects were 98.35%, 96.65% and 93.33%, 96.65%. (5) Chuxiong cheek wasp indoor control experiments. The components were cottonseed shell: wheat bran: sawdust: glucose: solid medium of CaSO4=40:40:18:1:1, solid fermentation of SWYH06 strain. The results showed that the mycelium growth and sporulation of SWYH06 strain on this medium were good, 10~15d At 20d, the sporulation of solid fermentations reached 4.5 x 108 spores, /g~6.7 x 108 spores /g. were treated with 3 different kinds of spore application methods. The results showed that 2 methods of solid fermenting with solid fermenting and 2 methods of solid fermentation on the soil surface before the larva were applied to the soil were used to treat the larva of the flat leaf peak of Chuxiong cheek. After 3 months, the corrected mortality rate reached 74.08 + 7.64% and 63.33 + 12.58%, indicating that the SWYH06 strain has a good application prospect in the biological control of Chuxiong.

【學(xué)位授予單位】：西南林業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S763

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