中国农业科学 ›› 2019, Vol. 52 ›› Issue (14): 2450-2467.doi: 10.3864/j.issn.0578-1752.2019.14.006

• 植物保护 • 上一篇    下一篇

整合微生物组菌剂的提出、研发与应用

刘波1,陈倩倩1,王阶平1,阮传清1,陈燕萍1,夏江平2,车建美1,陈峥1,潘志针1,文笑2,朱育菁1,张海峰1,郑雪芳1   

  1. 1福建省农业科学院农业生物资源研究所,福州 350003
    2厦门江平生物基质技术股份有限公司,福建厦门 361009
  • 收稿日期:2019-03-19 接受日期:2019-05-09 出版日期:2019-07-16 发布日期:2019-07-26
  • 作者简介:刘波(通信作者),E-mail: fzliubo@163.com
  • 基金资助:
    国家重点研发计划(2018YFD0500201);福建省公益类科研院所专项(2018R1017-1);福建省公益性项目(2016R1017-2);福建省自然科学基金(2018J01036)

Proposition, Development and Application of the Integrated Microbiome Agent (IMA)

Bo LIU1,QianQian CHEN1,JiePing WANG1,ChuanQing RUAN1,YanPing CHEN1,JiangPing XIA2,JianMei CHE1,Zheng CHEN1,ZhiZhen PAN1,Xiao WEN2,YuJing ZHU1,HaiFeng ZHANG1,XueFang ZHENG1   

  1. 1Agricultural Bioresources Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003
    2Xiamen Jiangping Biology Substrate Technology Co. Ltd., Xiamen 361009, Fujian
  • Received:2019-03-19 Accepted:2019-05-09 Online:2019-07-16 Published:2019-07-26

摘要:

【目的】利用微生物发酵床作为发酵槽,猪粪氮素连续流加中温好氧发酵,将通过宏基因检测鉴定到的微生物组称为整合微生物组,生产高含菌量整合微生物组菌剂,作为植物病害生防菌剂。【方法】生产工艺:原料配制→发酵床发酵→猪粪氮素连续流加→好氧发酵控制→产品加工→产品包装等。生产技术:利用养猪使用1年以上的微生物发酵床,添加一层10 cm厚的30%豆饼粉+70%杏鲍菇菌糠垫料,每平方米1头猪作为猪粪氮素连续流加营养来源,每天翻耕1次,连续好氧发酵20 d后,取出上层20 cm的垫料,进入晾晒、粉碎、分筛、包装,加工成整合微生物组菌剂。【结果】整合微生物组菌剂产品技术指标:含水量29.74%,pH 7.56,有机质含量44.46%,全氮含量2.23%,腐殖酸含量11.20%,粗纤维含量14.06%,含菌量145×10 8cfu/g。宏基因组测定结果表明,菌剂样品序列(reads)条数平均值为99 701.75,每克菌剂含有细菌39门、96纲、189目、383科、786属、1 281种;其中,芽孢杆菌46种,9种为中国新记录种,即:①嗜气芽孢杆菌(Bacillus aerophilus)、②蚯蚓芽孢杆菌(Bacillus eiseniae)、③丝状芽孢杆菌(Bacillus filamentosus)、④柯赫芽孢杆菌(Bacillus kochii)、⑤根际芽孢杆菌(Bacillus rhizosphaerae)、⑥长型赖氨酸芽孢杆菌(Lysinibacillus macroides)、⑦淤泥大洋芽孢杆菌(Oceanobacillus caeni)、⑧拾蛤鸟氨酸芽孢杆菌(Ornithinibacillus scapharcae)、⑨海洋枝芽孢杆菌(Virgibacillus oceani);未发现猪细菌病原。可培养法分离的芽孢杆菌活菌数2.062×10 8cfu/g,宏基因组测定结果中,芽孢杆菌的总丰度为1.42%,以此推算菌剂有效细菌总含量145×10 8cfu/g。整合微生物组菌剂浸出液处理组的绿豆发芽率为96.67%,与清水对照组无显著差异(P>0.05),但胚根长比对照增加了58.08%。用5%—10%的菌剂配制成育苗基质,番茄出苗率提高了3.0%,株高增加了25.1%,对番茄青枯病的校正防治效果可达79.41%。整合微生物组菌剂的产品质量标准参考农业农村部生物有机肥标准(NY884-2012),初步确定为:有机质≥40%,含水量≤30%,pH 5.5—7.5,粪大肠菌群数≤100个/g,蛔虫卵死亡率>95%,有效期>6个月;重金属含量满足标准要求:砷<15 mg·kg -1,镉≤15 mg·kg -1,铅≤15 mg·kg -1,铬≤15 mg·kg -1,汞≤15 mg·kg -1;有效活菌数调整为:总细菌数≥30×10 8cfu/g,其中芽孢杆菌≥2×10 8cfu/g。 【结论】提出了整合微生物组菌剂的概念和产品技术标准。研发的整合微生物菌剂可促进种子根部生长,并对番茄青枯病有良好的防治效果。

关键词: 整合微生物组菌剂, 生物有机肥, 绿豆, 发芽率, 胚根长, 生长率, 青枯病, 防治效果

Abstract:

【Objective】Using microbial fermentation bed as fermentation tank, pig manure nitrogen was continuously added to achieve aerobic fermentation in medium temperature. Microbiome with high bacterial content identified by metagenomic technology was named as integrated microbiome. Integrated microbiome agent (IMA) was produced as plant disease biocontrol agent.【Method】The production process is as follows: material preparation→fermentation→nitrogen feeding→aerobic fermentation control→product processing→product packaging, etc. The fully manufacturing processes of IMAs were as follows: the litters consisting of soybean cake powders (30%) and Pleurotus eryngii mushroom substrates (70%) were added onto the pig microbial fermentation bed that had been used for more than one year with 10 cm thick; the aerobic fermentation was conducted for 20 days by ploughing the litters mixing with pig manures one time per day; then, the upper 20 cm litters were removed to produce IMAs by drying, crushing, screening and packaging.【Result】The physicochemical properties of the IMA products were moisture content 29.74%, pH 7.56, organic matter content 44.46%, total nitrogen content 2.23%, humic acid content 11.20%, crude fiber content 14.06%, and the amount of bacteria was 145×10 8cfu/g. The results of metagenomic determination showed that the average number of the short sequences (reads) was 99 701.75 in the IMA samples, and the IMA samples contained 39 phyla, 96 classes, 189 orders, 383 families, 786 genera, and 1 281 species of bacteria per gram. Among bacteria, 46 Bacillus-like species were detected, and 9 species were the new records in China, namely: (1) Bacillus aerophilus, (2) Bacillus eiseniae, (3) Bacillus filamentosus, (4) Bacillus kochii, (5) Bacillus rhizosphaerae, (6) Lysinibacillus macroides, (7) Oceanobacillus caeni, (8) Ornithinibacillus scapharcae, (9) Virgibacillus oceani. Importantly, no bacterial pathogen of pig was found in the IMA products. The total content of the cultural Bacillus-like species was 2.062×10 8cfu/g, and they accounted for 1.42% of total bacterial abundance in the metagenomic data. Therefore, the total amount of effective bacteria was estimated to be 145×10 8cfu/g in the IMA products based on the cultural Bacillus-like species content and total abundance of metagenomic detecting. The germination rate of mung bean in the IMA group was 96.67%, which was not significantly different from that in the clear water control group (P>0.05), but the radicle length was 58.08% higher than that in the control group. When the seedling-raising substrate was mixed with 5%-10% of IMAs, the emergence rate and plant height of tomato were increased by 3.0% and 25.1%, respectively, and the corrected control efficacy on tomato bacterial wilt was 79.41%. According to the national quality standards for the bio-organic fertilizers (NY884-2012), those of the IMA products were initially set as followings: organic matter content≥40%, moisture content≤30%, pH 5.5-7.5, fecal coliform count≤100 cfu/g, mortality of ascarid egg>95%, expiry date>6 months; The contents of heavy metals: As<15 mg·kg -1, Cd≤15 mg·kg -1, Pb ≤15 mg·kg -1, Cr≤15 mg·kg -1, and Hg≤15 mg·kg -1; The amount of living bacteria was adjusted to the total living bacteria amount≥30×10 8cfu/g, Bacillus-like species amount≥2×10 8cfu/g.【Conclusion】The concept and product technical standard of the integrated microbiome agents (IMAs) were proposed. The IMA can promote the growth of seed root and have a good control efficacy on tomato bacterial wilt.

Key words: integrated microbiome agent (IMA), bio-organic fertilizer, mung bean, germination rate, radicle length, growth rate, bacterial wilt, control efficacy