化肥减量配施中药源植物生长调节剂对当归质量和根际土壤细菌群落的影响

doi:10.13560/j.cnki.biotech.bull.1985.2021-0467

摘要/Abstract

摘要：

为实现当归种植中“化肥农药减量增效”的绿色发展目标,探讨化肥减量配施一种中药源植物生长调节剂对当归质量和根际土壤细菌群落的影响,为该中药源调节剂改善当归质量的机理提供科学依据。以当归为研究对象,设置常规量化肥（CK）、中药源调节剂配施化肥（T1）、中药源调节剂配施80%化肥（T2）、中药源调节剂配施60%化肥（T3）4个处理,测定当归5个生长期的生长指标、病情指标、产量、根际土壤理化指标等,并通过16S rDNA扩增子测序分析根际土壤细菌群落的变化。结果表明：（1）生长早期T1组和T2组的当归根长、芦头直径、地下生物量显著高于其他组,成药期CK组当归根部蚜虫出现率、蚜虫密度、病情等级、发病率均高于其他组,且产量最低,T2组产量最高,比CK组增产2.61倍;（2）全生长期当归根际土壤盐分、铵态氮含量、有效钾含量在组间差异不显著。有机质含量仅在苗期的CK组中高于其他组,其他时期组间无差异。有效磷含量在全生长期组间变化较大,pH值随着化肥减量逐渐增加;（3）全生长期当归根际土壤细菌群落多样性在组间无差异,门水平上群落结构组成一致,变形菌门（Proteobacteria）、拟杆菌门（Bacteroidetes）、厚壁菌门（Firmicutes）、放线菌门（Actinobacteria）、芽单胞菌门（Gemmatimonadetes）、酸杆菌门（Acidobacteria）为优势菌群;（4）属水平优势菌群为黄杆菌属（Flavobacterium）、鞘氨醇单胞菌属（Sphingomonas）、假单胞菌属（Pseudomonas）、拟杆菌属（Bacteroides）、MND1属、鞘氨醇杆菌属（Pedobacter）、柠檬酸杆菌属（Citrobacter）、Ellin6067属、芽单胞菌属（Gemmatimonas）、马赛菌属（Massilia）、鞘脂菌属（Sphingobium）、溶杆菌属（Lysobacter）、Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium属、Haliangium属和硝化螺旋菌属（Nitrospira）。其中假单胞菌属、柠檬酸杆菌属、Ellin6067属和马赛菌属丰度在当归不同生长时期存在组间显著差异,但仅有假单胞菌属丰度与当归根部生长指标显著正相关,与病情指标显著负相关,且CK组的假单胞菌属在早期和成药期较其他组显著降低。化肥减量配施中药源植物生长调节剂可促进当归早期根部生长,改善成药期根腐病病情,提高产量。该中药源调节剂通过增加当归根际土壤中假单胞菌属丰度促进当归质量的提升。

关键词: 当归, 植物生长调节剂, 16S rDNA, 细菌群落, 根际土壤

Abstract:

In order to achieve the green development goal of “reducing chemical fertilizer and increasing efficiency” in the cultivation of Angelica sinensis, the effects of chemical fertilizer reduction and application of plant growth regulators from traditional Chinese medicine on the quality and bacterial community in rhizosphere soil of A. sinensis were investigated, so as to provide scientific basis for the mechanism of improving the quality of A. sinensis. A. sinensis was taken as the research object. Four treatments were set up, including chemical fertilizer（CK）, Chinese medicine source regulator combined with chemical fertilizer（T1）, Chinese medicine source regulator combined with 80% chemical fertilizer（T2）,and Chinese medicine source regulator combined with 60% chemical fertilizer（T3）. The growth indexes,disease indexes,yield,rhizosphere soil physical and chemical indexes of A. sinensis in the 5 growth stages were determined,and the changes of bacterial community in the rhizosphere soil were analyzed by 16S rDNA amplification sequence. Results showed as below：1）The root length,main root diameter and root biomass of A. sinensis in treatment T1 and T2 were significantly higher than those in other treatments at early growth stage. The aphid emergence rate, aphid density,disease grade and incidence rate of A. sinensis in CK were higher than those in other treatments, and the yield was the lowest in CK,the highest in T2, which was 2.61 times higher than that of CK. 2）There was no significant difference in the salinity,ammonium nitrogen and available potassium among the treatments in whole life growth period. The organic matter in CK at seedling stage was higher than that in other treatments, but there was no difference among treatments at other growth stages. The available phosphorus changed greatly in the whole growth period, and the pH value increased gradually with the reduction of chemical fertilizer. 3）There was no difference in the diversity of bacterial community in the rhizosphere soil of A. sinensis during the whole growth period. The community structure was consistent at phylum level. Proteobacteria, Bacteroidetes, Firmicutes,Actinobacteria, Gemmatimonadetes and Acidobacteria were the dominant phylum. 4）At the genus level, the dominant genera were Flavobacterium, Sphingomonas, Pseudomonas, Bacteroides, MND1, Pedobacter, Citrobacter, Ellin6067, Gemmatimonas, Massilia, Sphingobiu, Lysobacter, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Haliangium, and Nitrospira. The abundance of Pseudomonas, Citrobacter, Ellin6067 and Massilia were significantly different among treatments in different growth stages of A. sinensis. However,only the abundance of Pseudomonas was positively correlated with the root growth index of A. sinensis,and negatively correlated with the disease index. The abundance of Pseudomonas in CK was significantly lower than that in other treatments in the early stage and drug preparation stage. Chemical fertilizer reduction and application of plant growth regulators from traditional Chinese medicine may promote the root growth of A. sinensis at early growth stages,improve the disease condition of root rot in the proprietary period,and increase the yield. The Chinese medicine source regulator may improve the quality of A. sinensis by increasing the abundance of Pseudomonas in the rhizosphere soil.

Key words: Angelica sinensis, plant growth regulator, 16S rDNA, bacterial community, rhizosphere soil

谢田朋, 柳娜, 刘越敏, 曲馨, 薄双琴, 景明. 化肥减量配施中药源植物生长调节剂对当归质量和根际土壤细菌群落的影响[J]. 生物技术通报, 2022, 38(3): 79-91.

XIE Tian-peng, LIU Na, LIU Yue-min, QU Xin, BO Shuang-qin, JING Ming. Effects of Chemical Fertilizer Reduction and Application of Plant Growth Regulators from Traditional Chinese Medicine on the Quality and Its Bacterial Community in Rhizosphere Soil[J]. Biotechnology Bulletin, 2022, 38(3): 79-91.

图/表 11

参考文献 29

[1]	李文涛, 杨世海. 当归质量评价研究现状[J]. 药物分析杂志, 2013, 33(3):517-523.
	Li WT, Yang SH. Current situation of Angelica sinensis quality evaluation[J]. Chin J Pharm Anal, 2013, 33(3):517-523.
[2]	顾志荣, 师富贵, 王永琦, 等. 不同产地当归药材质量研究进展[J]. 甘肃中医学院学报, 2014, 31(5):80-83.
	Gu ZR, Shi FG, Wang YQ, et al. Research progress on quality of Angelica sinensis from different place of production[J]. J Gansu Coll Tradit Chin Med, 2014, 31(5):80-83.
[3]	谢田朋, 柳娜, 王雅莉, 等. 不同产地当归品质的研究进展[J]. 中医药学报, 2020, 48(1):72-75.
	Xie TP, Liu N, Wang YL, et al. Quality research progress of Angelica sinensis from different regions[J]. Acta Chin Med Pharmacol, 2020, 48(1):72-75.
[4]	毛妍婷, 刘宏斌, 陈安强, 等. 长期施用有机肥对减缓菜田耕层土壤酸化的影响[J]. 生态环境学报, 2020, 29(9):1784-1791.
	Mao YT, Liu HB, Chen AQ, et al. Effects of long-term application of organic fertilizers on reducing soil acidification of plough layer in vegetable fields[J]. Ecol Environ Sci, 2020, 29(9):1784-1791.
[5]	邹湘, 易博, 张奇春, 等. 长期施肥对稻田土壤微生物群落结构及氮循环功能微生物数量的影响[J]. 植物营养与肥料学报, 2020, 26(12):2158-2167.
	Zou X, Yi B, Zhang QC, et al. Effects of long-term fertilization on the microbial community structure and the population of N cycle-related functional microorganism in paddy soil[J]. J Plant Nutr Fertil, 2020, 26(12):2158-2167.
[6]	李虹, 李汀贤, 赵凤亮, 等. 香蕉枯萎病发生区域土壤改良:间作对热带土壤微生物区系和pH相关关系的影响[J]. 园艺与种苗, 2017, 37(9):21-27.
	Li H, Li TX, Zhao FL, et al. Soil improvements in banana Fusarium wilt incidence areas:spatial variations and correlations of microbial flora and pH variables in tropical red soils[J]. Hortic Seed, 2017, 37(9):21-27.
[7]	肖婉君. 有机肥和减量化肥对当归成药栽培影响机理的研究[D]. 兰州:甘肃农业大学, 2020.
	Xiao WJ. Study of influence mechanism of organic fertilizer and reduced chemical fertilizer on Angelica sinensis medicinal formation cultivation[D]. Lanzhou:Gansu Agricultural University, 2020.
[8]	王文丽, 李娟, 赵旭. 生物有机肥对连作当归根际土壤细菌群落结构和根腐病的影响[J]. 应用生态学报, 2019, 30(8):2813-2821. pmid: 31418207
	Wang WL, Li J, Zhao X. Effects of biological organic fertilizer on rhisosphere soil bacteria community and root rot diseases of continuous cropping Angelica sinensis[J]. Chin J Appl Ecol, 2019, 30(8):2813-2821. doi: 10.13287/j.1001-9332.201908.030 pmid: 31418207
[9]	张奇茹, 谢英荷, 李廷亮, 等. 有机肥替代化肥对旱地小麦产量和养分利用效率的影响及其经济环境效应[J]. 中国农业科学, 2020, 53(23):4866-4878.
	Zhang QR, Xie YH, Li TL, et al. Effects of organic fertilizers replacing chemical fertilizers on yield, nutrient use efficiency, economic and environmental benefits of dryland wheat[J]. Sci Agric Sin, 2020, 53(23):4866-4878.
[10]	魏晓兰, 吴彩姣, 孙玮, 等. 减量施肥条件下生物有机肥对土壤养分供应及小白菜吸收的影响[J]. 水土保持通报, 2017, 37(1):40-44.
	Wei XL, Wu CJ, Sun W, et al. Effect of bio-organic fertilizer on soil nutrient supply and absorption of Bok choy under different decreasing fertilization[J]. Bull Soil Water Conserv, 2017, 37(1):40-44.
[11]	白雪纯, 张君红, 冯魁亮, 等. 化肥减量配施有机肥对青贮玉米产量、营养价值及土壤微生物活性的影响[J]. 草业科学, 2020, 37(2):348-354.
	Bai XC, Zhang JH, Feng KL, et al. Effects of chemical fertilizer reduction and application of organic manure on the yield and nutritive value of Zea mays and soil microbial activity[J]. Pratacultural Sci, 2020, 37(2):348-354.
[12]	于会丽, 徐国益, 徐变变, 等. 施用生物菌肥对桃园土壤养分及微生物功能多样性的影响[J]. 干旱地区农业研究, 2020, 38(6):91-97.
	Yu HL, Xu GY, Xu BB, et al. Effects of bio-fertilizer on soil nutrients and microbial functional diversity in peach orchard[J]. Agric Res Arid Areas, 2020, 38(6):91-97.
[13]	茹淑华, 徐万强, 侯利敏, 等. 连续施用有机肥后重金属在土壤-作物系统中的积累与迁移特征[J]. 生态环境学报, 2019, 28(10):2070-2078.
	Ru SH, Xu WQ, Hou LM, et al. Effects of continuous application of organic fertilizer on the accumulation and migration of heavy metals in soil-crop systems[J]. Ecol Environ Sci, 2019, 28(10):2070-2078.
[14]	郑茗月, 李海梅, 赵金山, 等. 微生物肥料的研究现状及发展趋势[J]. 江西农业学报, 2018, 30(11):52-56.
	Zheng MY, Li HM, Zhao JS, et al. Current situation and developmental trend of microbial fertilizer researches[J]. Acta Agric Jiangxi, 2018, 30(11):52-56.
[15]	柳娜, 张旭东, 王雅莉, 等. “中医农业”模式在当归种植中的作用研究[J]. 中兽医医药杂志, 2019, 38(5):19-21.
	Liu N, Zhang XD, Wang YL, et al. The effect of traditional Chinese compound pesticide and traditional Chinese compound fertilizer on the yield of Angelica Sinensis[J]. J Tradit Chin Vet Med, 2019, 38(5):19-21.
[16]	柳娜, 张夙萍, 谢田朋, 等. 肥药伴侣对岷归重金属元素含量的影响研究[J]. 中兽医医药杂志, 2020, 39(2):73-75.
	Liu N, Zhang SP, Xie TP, et al. Study on the effect of the companion of fertilizer and pesticideon on the content of heavy metals in Angelica sinensis from Min County[J]. J Tradit Chin Vet Med, 2020, 39(2):73-75.
[17]	柳娜, 王雅莉, 张旭东, 等. 中药复方农药与中药复方肥料对当归质量的影响研究[J]. 中药材, 2019, 42(2):267-270.
	Liu N, Wang YL, Zhang XD, et al. Effect of traditional chinese medicine compound pesticide and fertilizer on the quality of Angelica sinensis[J]. J Chin Med Mater, 2019, 42(2):267-270.
[18]	辛中尧, 徐红霞, 陈秀蓉. 枯草芽孢杆菌(Bacillus subtilis)B1、B2菌株对当归、黄芪的防病促进生长效果[J]. 植物保护, 2008, 34(6):142-144.
	Xin ZY, Xu HX, Chen XR. Effect of Bacillus subtilis B1, B2 on Angelica sinensis and Astragalus membranaceus of preventing of disease and growth-promoting[J]. Plant Prot, 2008, 34(6):142-144.
[19]	鲁如坤. 土壤农业化学分析方法[M]. 北京: 中国农业科技出版社, 2000.
	Lu RK. Soil and agricultural chemistry analysis method[M]. Beijing: China Agriculture Scientech Press, 2000.
[20]	Bolger AM, Lohse M, Usadel B. Trimmomatic:a flexible trimmer for Illumina sequence data[J]. Bioinformatics, 2014, 30(15):2114-2120. doi: 10.1093/bioinformatics/btu170 URL
[21]	Reyon D, Tsai SQ, Khayter C, et al. FLASH assembly of TALENs for high-throughput genome editing[J]. Nat Biotechnol, 2012, 30(5):460-465. doi: 10.1038/nbt.2170 URL
[22]	Caporaso JG, Kuczynski J, Stombaugh J, et al. QIIME allows analysis of high-throughput community sequencing data[J]. Nat Methods, 2010, 7(5):335-336. doi: 10.1038/nmeth.f.303 pmid: 20383131
[23]	Rognes T, Flouri T, Nichols B, et al. VSEARCH:a versatile open source tool for metagenomics[J]. PeerJ, 2016, 4:e2584. doi: 10.7717/peerj.2584 URL
[24]	Wang Q, Garrity GM, Tiedje JM, et al. Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy[J]. Appl Environ Microbiol, 2007, 73(16):5261-5267. doi: 10.1128/AEM.00062-07 URL
[25]	Oksanen J, Kindt R, Legendre P, et al. The vegan package:Community Ecology Package, 2007.http://r-forge.r-project.org/projects/vegan/.
[26]	白洋, 钱景美, 周俭民, 等. 农作物微生物组:跨越转化临界点的现代生物技术[J]. 中国科学院院刊, 2017, 32(3):260-265.
	Bai Y, Qian JM, Zhou JM, et al. Crop microbiome:breakthrough technology for agriculture[J]. Bull Chin Acad Sci, 2017, 32(3):260-265.
[27]	徐敏, 叶凡, 赖先荣, 等. HPLC法测定白芥子饮片中白芥子苷的含量[J]. 中国药房, 2016, 27(18):2549-2551.
	Xu M, Ye F, Lai XR, et al. Content determination of sinalbin in Sinapis alba decoction piece by HPLC[J]. China Pharm, 2016, 27(18):2549-2551.
[28]	张鹏, 赵伟琼, 苗莉云, 等. 七叶一枝花根内生细菌多样性分析及促生菌的筛选[J]. 中南民族大学学报:自然科学版, 2020, 39(6):596-600.
	Zhang P, Zhao WQ, Miao LY, et al. Diversity analysis of endophytic bacteria in the roots of Paris Polyphylla and screening of growth-promoting bacteria[J]. J South Central Univ Natl:Nat Sci Ed, 2020, 39(6):596-600.
[29]	柴晓虹, 姚拓, 李录山, 等. 12株植物根际促生菌促生功能稳定性评价及鉴定[J]. 草原与草坪, 2020, 40(5):68-75.
	Chai XH, Yao T, Li LS, et al. Stability evaluation of growth promoting function and identification of 12 PGPR strains[J]. Grassland Turf, 2020, 40(5):68-75.

处理 Treatment	氮肥 Nitrogen fertilizer /（kg·m^-2）	磷肥 Phosphorus fertilizer /（kg·m^-2）	中药源调节剂 Regulator from traditional Chinese medicine /（kg·m^-2）
CK	4.5	3.7	0
T1	4.5	3.7	10
T2	3.6	2.9	10
T3	2.7	2.2	10

处理 Treatment	氮肥 Nitrogen fertilizer /（kg·m^-2）	磷肥 Phosphorus fertilizer /（kg·m^-2）	中药源调节剂 Regulator from traditional Chinese medicine /（kg·m^-2）
CK	4.5	3.7	0
T1	4.5	3.7	10
T2	3.6	2.9	10
T3	2.7	2.2	10

采收时期 Collection period	处理 Treatment	株高 Plant height	地上部生物量 Aboveground biomass	根长 Root length	芦头直径 Main root diameter	地下部生物量 Underground biomass
D1	CK	1.31±0.01a	1.94±0.03a	0.98±0.03c	0.63±0.03c	1.37±0.02bc
	T1	1.24±0.03ab	1.81±0.06ab	1.09±0.02ab	0.75±0.01ab	1.49±0.01ab
	T2	1.16±0.02b	1.73±0.05b	1.13±0.02a	0.78±0.02a	1.61±0.03a
	T3	1.12±0.02b	1.54±0.07c	1.03±0.02bc	0.69±0.01bc	1.34±0.04c
D2	CK	1.54±0.02a	2.80±0.07a	0.86±0.03b	0.71±0.04b	1.46±0.06b
	T1	1.49±0.02a	2.69±0.07a	0.86±0.05b	0.78±0.06a	1.83±0.12a
	T2	1.36±0.02b	2.22±0.05c	1.02±0.04a	0.77±0.02a	2.05±0.08a
	T3	1.37±0.01b	2.45±0.04b	0.88±0.03b	0.74±0.03b	1.82±0.10a
D3	CK	1.68±0.01a	3.67±0.03a	1.15±0.02a	1.12±0.02a	2.65±0.05b
	T1	1.67±0.01a	3.49±0.03b	1.15±0.01a	1.12±0.02a	2.71±0.05b
	T2	1.65±0.01a	3.46±0.05b	1.13±0.02a	1.13±0.01a	2.89±0.03a
	T3	1.59±0.01b	3.26±0.05c	1.09±0.03a	1.12±0.02a	2.47±0.06c
D4	CK	1.66±0.01b	3.79±0.03abc	1.18±0.01a	1.14±0.01a	3.06±0.03a
	T1	1.70±0.01a	3.90±0.05a	1.18±0.01a	1.15±0.03a	3.07±0.05a
	T2	1.64±0.01b	3.69±0.04c	1.18±0.01a	1.09±0.02a	2.94±0.04a
	T3	1.66±0.01b	3.77±0.04b	1.15±0.02a	1.09±0.02a	2.96±0.05a
D5	CK	1.65±0.01c	3.75±0.04b	1.27±0.02a	1.22±0.02a	3.26±0.04a
	T1	1.71±0.01a	3.97±0.05a	1.31±0.01a	1.26±0.02a	3.34±0.04a
	T2	1.70±0.01a	4.01±0.05a	1.31±0.02a	1.27±0.02a	3.36±0.04a
	T3	1.68±0.01b	3.90±0.06a	1.29±0.02a	1.21±0.03a	3.26±0.06a

采收时期 Collection period	处理 Treatment	株高 Plant height	地上部生物量 Aboveground biomass	根长 Root length	芦头直径 Main root diameter	地下部生物量 Underground biomass
D1	CK	1.31±0.01a	1.94±0.03a	0.98±0.03c	0.63±0.03c	1.37±0.02bc
	T1	1.24±0.03ab	1.81±0.06ab	1.09±0.02ab	0.75±0.01ab	1.49±0.01ab
	T2	1.16±0.02b	1.73±0.05b	1.13±0.02a	0.78±0.02a	1.61±0.03a
	T3	1.12±0.02b	1.54±0.07c	1.03±0.02bc	0.69±0.01bc	1.34±0.04c
D2	CK	1.54±0.02a	2.80±0.07a	0.86±0.03b	0.71±0.04b	1.46±0.06b
	T1	1.49±0.02a	2.69±0.07a	0.86±0.05b	0.78±0.06a	1.83±0.12a
	T2	1.36±0.02b	2.22±0.05c	1.02±0.04a	0.77±0.02a	2.05±0.08a
	T3	1.37±0.01b	2.45±0.04b	0.88±0.03b	0.74±0.03b	1.82±0.10a
D3	CK	1.68±0.01a	3.67±0.03a	1.15±0.02a	1.12±0.02a	2.65±0.05b
	T1	1.67±0.01a	3.49±0.03b	1.15±0.01a	1.12±0.02a	2.71±0.05b
	T2	1.65±0.01a	3.46±0.05b	1.13±0.02a	1.13±0.01a	2.89±0.03a
	T3	1.59±0.01b	3.26±0.05c	1.09±0.03a	1.12±0.02a	2.47±0.06c
D4	CK	1.66±0.01b	3.79±0.03abc	1.18±0.01a	1.14±0.01a	3.06±0.03a
	T1	1.70±0.01a	3.90±0.05a	1.18±0.01a	1.15±0.03a	3.07±0.05a
	T2	1.64±0.01b	3.69±0.04c	1.18±0.01a	1.09±0.02a	2.94±0.04a
	T3	1.66±0.01b	3.77±0.04b	1.15±0.02a	1.09±0.02a	2.96±0.05a
D5	CK	1.65±0.01c	3.75±0.04b	1.27±0.02a	1.22±0.02a	3.26±0.04a
	T1	1.71±0.01a	3.97±0.05a	1.31±0.01a	1.26±0.02a	3.34±0.04a
	T2	1.70±0.01a	4.01±0.05a	1.31±0.02a	1.27±0.02a	3.36±0.04a
	T3	1.68±0.01b	3.90±0.06a	1.29±0.02a	1.21±0.03a	3.26±0.06a

处理 Treatment	蚜虫出现率 Aphid emergence rate /%	蚜虫密度 Aphid density /（Aphids·cm^-2）	病情指数 Disease index/%	发病率 Incidence rate/%	产量 Yield /（kg·m^-2）
CK	84.0±7.4a	12.20±1.39a	43.22±1.50a	88.0±6.6a	1.85±0.158c
T1	52.0±10.1b	11.44±1.60a	25.37±1.32b	52.2±10.1b	2.88±0.139b
T2	40.1±10.0b	11.96±1.53a	24.98±1.57b	48.0±19.1b	4.83±0.260a
T3	48.2±10.0b	11.36±1.62a	22.77±1.42b	40.1±10.0b	3.01±0.085b