张声峰 陈硕昌 梁文凤 严祥 冯艺峰 李桂珍 杨辉

摘要：【目的】从自然状态下的油茶壳堆肥产品中筛选功能芽孢杆菌，为油茶壳堆肥高效生产、饲料发酵等应用提供菌剂。【方法】利用高通量测序技术测定广西某油茶壳堆肥中微生物群落分布，筛选适宜堆肥微生物生长的培养基以获得多样性较丰富的堆肥芽孢杆菌菌群，经平板稀释涂布法分离纯化菌株，使用水解圈法和酶活测定方法进行水解酶功能分析，利用菌株形态特征观察、16S rDNA分子鉴定法明确菌株的种属，并运用分子生物学软件MEGAX构建系统发育树。使用重铬酸钾法测定腐殖酸含量。【结果】芽孢杆菌科（Bacillaceae）是优势菌科，占比达55.58%;分离到15株芽孢杆菌，同时具有淀粉酶、纤维素酶、蛋白酶3种水解酶活性的菌株有6株;同时具有淀粉酶和纤维素酶活性的有1株;仪具有蛋白酶或纤维素酶一种酶活性的有2株。其中菌株Bacillus sp.YXI1的蛋白酶活力达到27 07±3.28 U-mL-1，淀粉酶活力达123 97±3.19 U-mL-1，纤维素酶活力达15 75±0.23 U-mL-1。含有这15株菌的复合菌制剂有利于腐殖酸的生成，提高了堆肥品质。【结论】鉴定出的Bacillus cereus YX02和Bacillusflexus FYF01等菌株具有進一步研究丌发的价值，可用于开发油茶壳堆肥微生物菌制剂。

关键词：油茶壳堆肥;芽孢杆菌;功能性;筛选;应用

中图分类号：X 712;X53

文献标志码：A

文章编号：1008-03 84（ 2021） 07-0843-12

Functional Bacillus Species in Camellia Seed Shell Compost

ZHANGShengfeng 1， CHENShuochang 1， LIANGWenfeng 1， YANXiang 1. FENGYifeng 1， LIGuizhen 2. YANGHui 1*

（ I. College ofLife Science and Technology， Guangxi UniTJersity. Nanning， Guangxi 530004. China;

2. Guangxi Academy ofForestry Sciences， Nanning， Guangxi 530000， China ）

Abstract：【Objective】Bacillus spp that contribute to the fermentation of Camellia oleifera seed shells were isolated foreffective composting of the waste material. 【Method】 The microbial community in natural camellia seed shell compostfound in Guangxi was studied using the high-throughput sequencing technology. Suitable culture medium to foster the growthof richly diverse Bacillus spp from the compost was selected. Flora isolation by dilution with a streaking plate methodfollowed. The hydrolase activities of the isolates were determined by using the hydrolysis circle method and enzyme activityanalysis， the species identified by a 16S rDNA analysis， and the phylogeny constructed by MEGAX. The content of humic acidin the compost was measured by a potassium dichromate method. 【Result】 Bacillaceae was the dominant family in thecompost. It accounted for 55.58% of all isolated flora. Among the 15 Bacillus isolates. 6 exhibited activities of amylase，cellulase， and protease， one of amylase and cellulase. and 2 0f protease or protease. Strain YXI I showed a protease activity of27.07+3.28 U·mL-1. an amylase activity of 123.971- 3.19 U·mL-1. and a cellulase activity of 15.75+0.23 U·mL-1 In thepresence of numerous Bacillus spp that secreted varieties of hydrolases， formation of humic acid in the compost was enhanced.【Conclusion】 Some of the isolated strains. such as B. cereus YX02 and B. flexus FYFOI， might warrant further investigationto develop microbial inoculants for efficient composting camellia seed shells.

Key words： camellia seed shell compost; bacillus; functional; screening; application

0 引言

【研究意義】油茶是我国特有的木本食用油料树种。油茶壳是油茶果实加工茶油的副产品，随着油茶产业的发展，每年都有大量油茶壳产生。油茶壳占整个油茶果实质量的60%以上，其主要成分是纤维素、半纤维素、木质素[1]，已经在有机肥料、钾盐、活性炭的生产中得到利用，具有广阔的发展空间与市场空间‘纠。……