本文選題：銀杏 + 內(nèi)生真菌��； 參考：《福建農(nóng)林大學(xué)》2015年碩士論文

【摘要】：植物內(nèi)生真菌(Endophytic Fungus)與植物長(zhǎng)期協(xié)同進(jìn)化的過(guò)程中能夠產(chǎn)生與宿主植物相同或相似的活性物質(zhì),植物內(nèi)生真菌在農(nóng)業(yè)和醫(yī)藥研究上已得到廣泛應(yīng)用。本論文從傳統(tǒng)的中藥材植物銀杏(Ginkgo biloba)中分離篩選具殺褐飛虱(Nilaparvata lugens(Stal))活性的植物內(nèi)生真菌為主線(xiàn),為下一步分離鑒定活性物質(zhì)提供材料。主要結(jié)果如下：1.從銀杏葉、短枝和根部共分離出內(nèi)生真菌172株。其中,短枝中分離得到內(nèi)生真菌最多,達(dá)78株,占分離總數(shù)的45.35%；根部次之,占總數(shù)的38.37%；而葉片中僅分離出28株菌,占總數(shù)的16.28%。2.以褐飛虱作為靶標(biāo)昆蟲(chóng),采用點(diǎn)背法進(jìn)行具殺蟲(chóng)活性?xún)?nèi)生真菌初步篩選,結(jié)果發(fā)現(xiàn)有FC-5-3、FP-8-1、JP-6-2、JP-5-1、JP-11、JP-6-5、NP-5-3、NP-2-2、 NC-3-3等9株內(nèi)生真菌具有較好的觸殺活性。其中活性最好的JP-6-2菌株48h校正死亡率高達(dá)97.69%,LCso為0.44mg/ml。3.為了明確JP-6-2菌株代謝產(chǎn)物中殺蟲(chóng)活性物質(zhì)的理化性質(zhì),測(cè)定了該物質(zhì)在紫外光照、高溫及室外環(huán)境條件的穩(wěn)定性,測(cè)試結(jié)果表明：該殺蟲(chóng)活性物質(zhì)在高溫下比較穩(wěn)定,具有較好的耐熱性,但在紫外光、室外環(huán)境條件下穩(wěn)定較差。4.通過(guò)對(duì)該菌株進(jìn)行菌絲體、孢子形態(tài)和ITS鑒定,表明JP-6-2菌株屬于鏈格孢屬(Altermaria.sp),可能屬于細(xì)極鏈格孢種(Altermaria tenuissima),并構(gòu)建了系統(tǒng)發(fā)育樹(shù)研究其種屬地位。5.論文通過(guò)考察多個(gè)單因素和采用正交實(shí)驗(yàn)方法來(lái)優(yōu)化菌株發(fā)酵條件,最終優(yōu)化結(jié)果是：蔗糖30.00g/L,尿素2.00g/L,磷酸氫二鉀4.00g/L,硫酸鎂1.00g/L,硫酸亞鐵0.01g/L氯化鉀0.50g/L,每搖瓶裝液量50.0ml/250ml,種子液接種量120.0ml/1L,溫度27.0℃,轉(zhuǎn)速200r/min,初始pH值8.5,發(fā)酵培養(yǎng)時(shí)間9d。粗提物重量達(dá)到0.456g/L,增加237.78%；LC50達(dá)到0.58mg/ml,增加31.82%。
[Abstract]:Endophytic Fungus (endophytic Fungus) can produce the same or similar active substances as host plants during long-term coevolution. Plant endophytic fungi have been widely used in agricultural and medical research. In this paper, endophytic fungi with the activity of Nilaparvata lugens (Stal) were isolated and screened from the traditional Chinese medicinal plant ginkgo biloba as the main line to provide materials for the further isolation and identification of active substances. The main results are as follows: 1. 172 strains of endophytic fungi were isolated from leaves, short branches and roots of Ginkgo biloba. The endophytic fungi isolated from the short branches were the most, accounting for 78 strains, accounting for 45.35% of the total, followed by the roots, accounting for 38.37% of the total, while only 28 strains were isolated from the leaves, accounting for 16.28.2% of the total. The primary screening of endophytic fungi with insecticidal activity was carried out using brown planthopper as target insect. The results showed that 9 endophytic fungi such as FC-5-3, FP-8-1, JP-6-1, JP-11JP-6-5, NP-5-3NP-2-2 and NC-3-3 had good contact killing activity. Among them, the best JP-6-2 strain had a 48h adjusted mortality of 97.69% LCso of 0.44 mg / ml. 3. In order to determine the physical and chemical properties of the insecticidal active substance in the metabolites of JP-6-2 strain, the stability of the substance in ultraviolet light, high temperature and outdoor environment was determined. The results showed that the insecticidal active substance was stable at high temperature. It has good heat resistance, but it is less stable in ultraviolet light and outdoor environment. The identification of mycelia, spore morphology and its showed that JP-6-2 belonged to Altermaria.sp, which may belong to (Altermaria tenuissima), and constructed phylogenetic tree to study its genus status. In this paper, the fermentation conditions of the strain were optimized by investigating several single factors and adopting the orthogonal experiment method. The optimized results were as follows: sucrose 30.00g / L, urea 2.00g / L, potassium hydrogen phosphate 4.00g / L, magnesium sulfate 1.00g / L, ferrous sulfate 0.01g / L potassium chloride 0.50g / L, inoculation amount of seed liquid 120.0ml / L, temperature 27.0 鈩，

本文編號(hào)：2070818