發(fā)布時間：2018-04-24 03:25

  本文選題：白腐真菌 + 漆酶��； 參考：《佳木斯大學(xué)》2017年碩士論文

【摘要】：隨著農(nóng)藥大面積的使用,如何降低殘留在環(huán)境中的農(nóng)藥成為現(xiàn)階段重要科學(xué)研究內(nèi)容,在多學(xué)科的研究中,生物降解無污染,廣泛性以及高效性成為主要技術(shù)之一。白腐真菌作為一種生物降解菌,其通過產(chǎn)生的各種酶系,主要是漆酶,對農(nóng)藥有著高效的降解效果。因此,如何確定產(chǎn)漆酶白腐真菌及提高其產(chǎn)漆酶量成為研究降解農(nóng)藥的關(guān)鍵。本實(shí)驗(yàn)研究了在佳木斯四豐山采集的白腐真菌,應(yīng)用愈創(chuàng)木酚培養(yǎng)基初步篩選出產(chǎn)漆酶的8種白腐真菌,進(jìn)行培養(yǎng)基的優(yōu)化,提高產(chǎn)漆酶量,應(yīng)用ABTS法測定漆酶活性。并且將篩選出產(chǎn)漆酶最多的4個菌種對東北溫室大棚土壤中農(nóng)藥殘留嘧菌酯的降解,應(yīng)用HPLC進(jìn)行檢測嘧菌酯殘留量計(jì)算降解率,結(jié)果表明:1、將采集的白腐真菌分離純化后接種到含愈創(chuàng)木酚的培養(yǎng)基上,能使培養(yǎng)基產(chǎn)生紅色或淡紫色的有東方栓菌、多毛栓菌、桑黃、輪紋韌革菌、樹舌、松生擬層孔菌、野生香菇、朱紅密孔菌,這8種白腐真菌都產(chǎn)漆酶。2、根據(jù)培養(yǎng)基營養(yǎng)因素和培養(yǎng)條件的篩選得出:葡萄糖和硝酸鉀是多數(shù)白腐真菌的碳氮源,金屬陽離子Cu2+、K+、Mg2+對白腐真菌產(chǎn)漆酶有促進(jìn)作用,而Ca2+對產(chǎn)酶有抑制作用,Zn2+對產(chǎn)漆酶沒有影響。溫度和pH對白腐真菌產(chǎn)漆酶有顯著影響,在酸性條下,溫度為25-30℃產(chǎn)漆酶量最佳。3、8種白腐真菌經(jīng)過培養(yǎng)基優(yōu)化后得出:東方栓菌產(chǎn)漆酶活性最高為133.14U/ml,是PDA培養(yǎng)基產(chǎn)漆酶活性的1.93倍;多毛栓菌產(chǎn)漆酶活性最高為73.05 U/ml,是PDA培養(yǎng)基產(chǎn)漆酶活性的1.50倍;桑黃產(chǎn)漆酶活性最高為31.48 U/ml,是PDA培養(yǎng)基產(chǎn)漆酶活性的1.85倍;輪紋韌革菌產(chǎn)漆酶活性最高為157.36 U/ml,是PDA培養(yǎng)基產(chǎn)漆酶活性的2.72倍;樹舌產(chǎn)漆酶活性最高為59.05 U/ml,是PDA培養(yǎng)基產(chǎn)漆酶活性的1.52倍;松生擬層孔菌產(chǎn)漆酶活性最高為127.17 U/ml,是PDA培養(yǎng)基產(chǎn)漆酶活性的1.70倍;野生香菇產(chǎn)漆酶活性最高為448.16 U/ml,是PDA培養(yǎng)基產(chǎn)漆酶活性的1.42倍;朱紅密孔菌產(chǎn)漆酶活性為137.77 U/ml,是PDA培養(yǎng)基產(chǎn)漆酶活性的2.36倍。4、將篩選出產(chǎn)漆酶最多的野生香菇、輪紋韌革菌、朱紅密孔菌、東方栓菌用最佳培養(yǎng)基進(jìn)行培養(yǎng),提取粗酶液,對采集自溫室大棚土壤中含殘留嘧菌酯農(nóng)藥進(jìn)行降解。結(jié)果表明:野生香菇的降解率最高,降解率達(dá)到了99.99%,輪紋韌革菌和朱紅密孔菌的降解率都達(dá)到了90%以上,分別為94.35%和93.93%,東方栓菌的降解率最低為82.63%。
[Abstract]:With the extensive use of pesticides, how to reduce the residue of pesticides in the environment has become an important scientific research content at this stage. In the multidisciplinary research, biodegradation without pollution, extensiveness and efficiency has become one of the main technologies. White rot fungi, as a biodegradable fungus, produce various enzyme systems, mainly laccase, which have high degradation effect on pesticide. Therefore, how to determine laccase-producing white rot fungi and increase their laccase production is the key to study the degradation of pesticides. The white rot fungi collected in Sifeng Mountain of Jiamusi were studied in this paper. Eight kinds of white rot fungi producing laccase were preliminarily screened by guaiacol medium. The laccase production was improved by optimizing the culture medium, and the laccase activity was determined by ABTS method. Four strains producing laccase were selected to degrade the pesticide residue in greenhouse soil in Northeast China, and the degradation rate was calculated by using HPLC to detect the residue of pyridamole. The results showed that when the collected white rot fungi were isolated and purified and inoculated on the medium containing guaiacol, they could produce red or lavender oriental suppository, polychaete, mulberry yellow, rotund leatherback, tree tongue and pine laminopore. Wild Lentinus edodes and Phyllostoma sinensis, all of which produce laccase. 2. According to the nutrient factors of culture medium and the screening of culture conditions, glucose and potassium nitrate are the carbon and nitrogen sources of most white rot fungi. Metal cationic Cu2 + K + Mg2 promoted laccase production by white rot fungi, while Ca2 inhibited laccase production. Zn2 had no effect on laccase production. The effects of temperature and pH on laccase production of white rot fungi were significant. The laccase production of 8 species of white rot fungi was optimized at 25 鈩，

本文編號：1794939