發(fā)布時間：2018-03-23 12:50

  本文選題：纖維素降解菌　切入點(diǎn)：復(fù)合菌劑　出處：《西北農(nóng)林科技大學(xué)》2015年碩士論文

【摘要】：隨著果品產(chǎn)業(yè)的大面積擴(kuò)展,每年修剪果樹枝條的量逐年增加,果樹枝條堆放占用大面積的土地,焚燒造成環(huán)境污染,還浪費(fèi)了寶貴的生物資源。本研究從堆肥產(chǎn)物和秦嶺山區(qū)的土壤樣品中分離純化高效纖維素降解菌株,將其組合培養(yǎng),構(gòu)建篩選出一組高效的纖維素混合菌群。并制成復(fù)合微生物菌劑應(yīng)用于果樹枝條和豬糞堆肥過程中,研究復(fù)合微生物菌劑對果樹枝條和豬糞堆肥過程中理化性質(zhì)、酶活性、微生物功能多樣性以及微生物遺傳多樣性的影響。主要結(jié)果表現(xiàn)在以下幾個方面:1、纖維素降解菌的篩選、鑒定及混合菌群的構(gòu)建通過剛果紅染色法初篩,再以內(nèi)切酶(CMC)、纖維素全酶(FPA)2種酶的六天酶活性為依據(jù)進(jìn)行復(fù)篩,得到5株酶活性較高的纖維素降解菌。經(jīng)生理生化測定和分子生物學(xué)鑒定,菌株2-3、10-2、3-5、6-2和10-4分別為羅爾斯通菌(Ralstoinia sp.)、青霉菌(Penicillium sp.)、黃灰青霉菌(Penicillium aurantioqriseum)、支頂孢菌(Acremonium alternatum)和毛殼霉菌(Chaetomium piluliferum)。對5株菌進(jìn)行組合培養(yǎng),得到最優(yōu)組合為10-2/3-5/2-3/6-2,其CMC、FPA和木聚糖酶活性分別為16.3、58.6和11083.7 U/m L,均比單菌株有一定程度提高。2、復(fù)合微生物菌劑對果樹枝條堆肥過程中酶活性的影響接種復(fù)合微生物菌劑在堆肥前期可以提高堆肥溫度,比對照處理高溫期(高于55℃)持續(xù)時間多3d,促進(jìn)堆料的腐熟。接種菌劑還能有效地提高果樹枝條堆肥過程中酶活性,脲酶、多酚氧化酶、纖維素酶、漆酶、錳過氧化物酶和木質(zhì)素過氧化物酶活性在堆肥過程中分別比CK處理提高2.3%~71.4%、0.3%~28.4%、15.0%~19.8%、1.0%~11.0%、4.1%~26.8%、4.0%~22.2%。3、復(fù)合微生物菌劑對果樹枝條堆肥過程中微生物多樣性的影響B(tài)iolog微平板法測定結(jié)果表明,接種復(fù)合菌劑顯著提高了堆肥中微生物的平均每孔顏色變化率(average well color development),并改變了高溫期和降溫期微生物對6大類碳源的利用;主成分分析表明,復(fù)合菌劑主要在高溫期發(fā)揮作用,對堆料中微生物起分異作用的碳源主要為糖類和氨基酸類。測序結(jié)果表明,好氧堆肥過程中優(yōu)勢細(xì)菌主要為芽孢桿菌屬(Bacillus sp.PML14)、不動桿菌屬(Pseudomonas sp.)、厚壁菌門屬(Firmicutes bacterium)、假單孢菌屬(Pseudomonas sp.)、席藻菌屬(Phormidium sp)和大量的用傳統(tǒng)方法不能培養(yǎng)但在堆肥過程中發(fā)揮了重要作用的細(xì)菌,其中Bacillus sp.、Uncultured bacterium isolate DGGE gel band、Phormidium sp.和Uncultured beta proteobacterium isolate DGGE gel band在堆肥的高溫期發(fā)揮了主要作用。接種菌劑處理能夠顯著提高堆肥過程的細(xì)菌群落多樣性,促進(jìn)細(xì)菌群落結(jié)構(gòu)的演替,加快堆肥的腐熟進(jìn)程。
[Abstract]:With the extensive expansion of fruit industry, the amount of pruning fruit tree branches increases year by year, and fruit tree branches pile up and occupy a large area of land, causing environmental pollution caused by incineration. High efficient cellulose degrading strains were isolated from compost products and soil samples from Qinling Mountain area and cultured in combination. A group of efficient cellulosic mixed bacteria was constructed and selected, and the compound microbial fungicides were used in the process of fruit tree branch and pig manure composting. The physicochemical properties and enzyme activity of the compound microbial fungicides on fruit tree branch and pig manure composting were studied. Effects of microbial functional diversity and microbial genetic diversity. The main results were as follows: 1. Screening and identification of cellulosic degrading bacteria and construction of mixed bacteria by Congo red staining. On the basis of six days enzyme activity of two enzymes, CMCX and FPAN, five strains of cellulose degrading bacteria with high enzyme activity were obtained, which were identified by physiological and biochemical tests and molecular biology. Strains 2-3, 10-2, 3-5, 6-2 and 10-4 were Ralstoinia sp., Penicillium sp., Penicillium aurantioqriseumum, Acremonium alternatum and Chaetomium piluliferum, respectively. The optimum combination was 10-2 / 3-5 / 2-3-3 / 6-2, and the CMCFPA and xylanase activities were 16.3U / 58.6 and 11083.7 U / mL, respectively, which were higher than that of single strain to some extent. The effect of compound microbial agents on enzyme activity in composting process of fruit trees was studied. At the early stage of composting, the composting temperature can be raised. Compared with the control treatment, the duration of high temperature period (above 55 鈩，

本文編號：1653532