波尔多液对烟草叶际微生物群落结构与代谢功能的影响

刘亭亭; 汪汉成; 孙美丽; 尹国英; 张盼; 向立刚; 蔡刘体; 孟建玉; 张长青

doi:10.16801/j.issn.1008-7303.2022.0087

波尔多液对烟草叶际微生物群落结构与代谢功能的影响

doi: 10.16801/j.issn.1008-7303.2022.0087

刘亭亭^{1, 2,},
汪汉成^1, ,,
孙美丽^{1, 2},
尹国英^1,,
张盼^1,,
向立刚^{1, 2},
蔡刘体¹,
孟建玉¹,
张长青²

1.
贵州省烟草科学研究院，贵阳 550081
2.
长江大学农学院，湖北荆州 434025

基金项目: 国家自然科学基金 (31960550，32160522)；贵州省科技基金项目 (黔科合基础-ZK[2021]重点036)；中国烟草总公司科技项目 (110202001035 (LS-04)，110202101048 (LS-08) )；贵州省“百层次”创新型人才 (黔科合平台人才-GCC[2022]028-1)

详细信息

作者简介:
刘亭亭，3026049684@qq.com

通讯作者:
汪汉成，xiaobaiyang126@hotmail.com

中图分类号: S482.2；X592
计量
- 文章访问数: 357
- HTML全文浏览量: 112
- PDF下载量: 37
- 被引次数: 0
出版历程
- 收稿日期: 2022-05-05
- 录用日期: 2022-08-22
- 网络出版日期: 2022-10-18
- 刊出日期: 2022-12-10

Effects of Bordeaux mixture on the community structure and metabolic function of tobacco phyllosphere microorganisms

LIU Tingting^{1, 2
,},
WANG Hancheng^{1
, ,},
SUN Meili^{1, 2},
YIN Guoying^1
,,
ZHANG Pan^1
,,
XIANG Ligang^{1, 2},
CAI Liuti¹,
MENG Jianyu¹,
ZHANG Changqing²

1.
Guizhou Academy of Tobacco Science, Guiyang 550081, China
2.
College of Agriculture, Yangtze University, Jingzhou 434025, Hubei Province, China

Funds: National Natural Science Foundation of China (31960550, 32160522); Guizhou Science and Technology Fund Project (The Basis of Guizhou Science and Technology Cooperation-ZK [2021] Key 036); Technology Project of China National Tobacco Corporation (110202001035 (LS-04), 110202101048 (LS-08)); Guizhou Province "100 Level" Innovative Talents (Qian Science Cooperation Platform Talents-GCC [2022] 028-1)

摘要

摘要: 测定了波尔多液对烟草赤星病菌的毒力，并采用高通量测序与Biolog代谢表型技术分别测定了其对烟叶健康与感病组织叶际微生物群落结构和代谢功能的影响。结果表明：波尔多液对烟草赤星病菌的抑制活性较弱，其抑制菌丝生长和孢子萌发的EC₅₀值分别为450.19和757.17 mg/L。健康与感病烟叶组织叶际细菌均分布于变形菌门 (6.93%和39.07%) 和厚壁菌门 (16.45%和0.65%)，优势细菌均有Kosakonia (3.46%和22.38%) 和假单胞菌属 (0.22%和5.95%)；真菌均分布于子囊菌门 (63.82%和93.74%) 和担子菌门 (6.82%和2.53%)，优势真菌有链格孢属 (36.48%和84.52%) 、Symmetrospora (5.56%和2.27%) 和枝孢霉属 (14.87%和6.66%)。波尔多液1 500 g/hm²处理对健康和感病烟叶叶际细菌和真菌群落结构与代谢功能均有影响，处理5 d时降低了叶际Kosakonia、鞘脂单胞菌属和乳杆菌属的相对丰度，增加了假单胞菌属、劳尔氏菌属等6种细菌菌属的相对丰度；降低了链格孢属、Symmetrospora等6种真菌属的相对丰度，增加了亚隔孢壳属、绿僵菌属等10种真菌属的相对丰度。处理10和15 d时对叶际真菌、细菌的影响逐渐降低。健康与感病烟叶叶际微生物均可高效代谢糖类、氨基酸类、羧酸类、双亲化合物、聚合物和胺/氨基化合物等29种碳源，但对α-丁酮酸的代谢较弱。波尔多液处理对烟叶叶际微生物的代谢抑制活性随时间延长逐渐减弱。研究结果揭示了波尔多液施用不同时期后对烟叶叶际微生物的影响规律，为了解药剂持效期的生态效益提供了参考依据。
- 烟草赤星病 /
- 波尔多液 /
- 叶际微生物 /
- 群落结构 /
- 微生物多样性 /
- 代谢功能
Abstract: In this paper, the virulence of Bordeaux mixture against Alternaria alternata was determined, and the effects of Bordeaux mixture on phyllosphere microbial community structure and metabolic function of healthy and diseased tobacco leaves were determined by high-throughput sequencing and Biolog metabolic phenotype technology. Results showed that Bordeaux mixture had a weak inhibitory activity against A. alternata with EC₅₀ values for inhibiting mycelial growth and spore germination of 450.19 and 757.17 mg/L, respectively. The phyllosphere bacteria in both healthy and diseased tobacco were distributed in Proteobacteria (6.93% and 39.07%) and Firmicutes (16.45% and 0.65%), and the dominant bacterial genera were Kosakonia (3.46% and 22.38%) and Pseudomonas (0.22% and 5.95%). The phyllosphere fungi in both healthy and diseased tobacco were distributed in Ascomycota (63.82% and 93.74%) and Basidiomycota (6.82% and 2.53%), and the dominant fungal genera were Alternaria (36.48% and 84.52%), Symmetrospora (5.56% and 2.27%) and Cladosporium (14.87% and 6.66%). Bordeaux mixture at 1 500 g/hm² affected the community structure and metabolic function of phyllosphere bacteria and fungi in healthy and diseased tobacco. In the bacterial and fungal communities, the relative abundances of Kosakonia, Sphinomonas, and Lactobacillus decreased, while the relative abundances of 6 bacterial genera including Pseudomonas, Ralstonia increased after 5 days of treatment. The relative abundance of 6 fungal genera including Alternaria, Symmetrospora decreased, while the relative abundance of 10 fungal genera including Didymella, Metarhizium increased after 5 days of treatment. The effects of Bordeaux mixture on phyllosphere fungi and bacteria gradually decreased after 10 and 15 days of treatment. Twenty-nine carbon sources including carbohydrates, amino acids, carboxylic acids, amphiphiles, polymers, and amine/amino compounds could be efficiently metabolized by phyllosphere microorganisms of healthy and diseased tobacco leaves, but the metabolism of α-butyric acid was weak. The metabolic inhibition activity of Bordeaux mixture on phyllosphere microorganisms decreased gradually with the increase of time. The results of the study revealed the regularity of the effect of Bordeaux mixture on the microbes of tobacco leaves after different periods of application and provided a reference for understanding the ecological benefits of the fungicide in the effective period.
- tobacco brown spot /
- Bordeaux mixture /
- phyllosphere microorganisms /
- community structure /
- microorganism diversity /
- metabolic function

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图 1 波尔多液对烟叶叶际细菌门 (A) 和属(B)水平上的群落组成的影响

Figure 1. The effects of Bordeaux mixture on the community composition of bacteria in the phyllosphere of tobacco at phyla(A) and genera(B) levels

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图 2 波尔多液处理对烟叶叶际真菌门 (A) 和属(B)水平上的群落组成的影响

Figure 2. The effects of Bordeaux mixture on the community composition of fungi in the phyllosphere of tobacco at phyla(A) and genera(B) levels

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图 3 波尔多液处理烟叶叶际细菌 (A) 和真菌(B)群落OTU分布花瓣图

Figure 3. Flower map of OTU distribution of bacterial(A) and fungal(B) communities in phyllosphere of tobacco treated with Bordeaux mixture

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图 4 波尔多液处理烟叶叶际微生物代谢功能的聚类热图

注：颜色值分别代表微生物代谢碳源的程度，0~100为代谢程度低，100~200为代谢程度一般，200以上为代谢程度高。

Figure 4. Cluster heat map of the effect of Bordeaux mixture on the metabolism of tobacco phyllosphere microorganism

Note: The color values represent the degree of the microbes metabolizing carbon sources. The value 0-100, 100-200, and >200 represent the leaf microbes metabolized in the Biolog ECO microplate poorly, moderately, and effectively, respectively.

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表 1 样品采集信息

Table 1. Samples information

取样部位 Sampling location		施药前 0 d 0 day before application	施药后 5 d 5 days after application	施药后 10 d 10 days after application	施药后 15 d 15 days after application
健康组织 Healthy tissue	样品编号 Sample No.	BEJ01	BEJ11	BEJ21	BEJ31
		BEJ02	BEJ12	BEJ22	BEJ32
		BEJ03	BEJ13	BEJ23	BEJ33
	分组编号 Group No.	BEJ0	BEJ1	BEJ2	BEJ3
感病组织 Diseased tissue	样品编号 Sample No.	BEB01	BEB11	BEB21	BEB31
		BEB02	BEB12	BEB22	BEB32
		BEB03	BEB13	BEB23	BEB33
	分组编号 Group No.	BEB0	BEB1	BEB2	BEB3

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表 2 波尔多液对烟草赤星病菌的毒力

Table 2. Toxicity test of Bordeaux mixture to Alternaria alternata

	质量浓度 Mass concentration/(mg/L)	抑制率 Inhibition ratio/%	EC₅₀/(mg/L)	EC₉₅/(mg/L)	回归方程 Regression equation	相关系数 r
菌丝生长 Mycelial growth	0	0	450.19	4987.45	y=4.26 + 1.61x	0.992
	100	15.99
	200	25.17
	400	49.37
	800	66.34
	1600	79.56
孢子萌发 Conidia germination	0	0	757.17	3108.01	y=7.67 + 2.67x	0.997
	200	6.74
	400	22.80
	800	49.97
	1600	82.41

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表 3 波尔多液对烟叶叶际微生物多样性的影响 (OTU水平)

Table 3. Effect of Bordeaux mixture on the diversity of tobacco phyllosphere microorganism (OTU level)

	分组 Group	香农指数 Shannon index	Chao1 指数 Chao1 index	ACE 指数 ACE index	覆盖度 Goods coverage
细菌 Bacteria	BEB0	1.24 ± 0.54 bc	20.00 ± 6.81 b	29.88 ± 10.97 b	0.98 ± 0.01 a
	BEJ0	1.26 ± 0.08 bc	46.44 ± 25.14 ab	41.69 ± 12.65 b	0.97 ± 0.01 a
	BEB1	2.35 ± 0.09 ab	46.33 ± 13.86 ab	48.09 ± 14.73 b	0.96 ± 0.01 a
	BEJ1	0.73 ± 0.19 c	71.75 ± 39.56 ab	74.87 ± 39.93 b	0.95 ± 0.02 ab
	BEB2	2.78 ± 1.11 a	97.54 ± 29.63 ab	120.30 ± 31.45 ab	0.90 ± 0.03 b
	BEJ2	1.53 ± 0.19 abc	130.98 ± 40.17 a	212.81 ± 45.16 a	0.90 ± 0.01 b
	BEB3	1.57 ± 0.14 abc	36.67 ± 13.32 b	48.14 ± 21.83 b	0.96 ± 0.02 a
	BEJ3	1.63 ± 0.50 abc	70.34 ± 43.99 ab	90.39 ± 55.60 b	0.95 ± 0.02 ab
真菌 Fungi	BEB0	0.99 ± 0.42 c	64.89 ± 6.18 e	70.36 ± 5.40 e	0.999 ± 0.000 a
	BEJ0	3.17 ± 0.16 b	193.26 ± 58.62 de	208.14 ± 66.06 de	0.998 ± 0.001 ab
	BEB1	1.37 ± 0.22 c	218.21 ± 42.63 d	253.44 ± 63.40 cd	0.997 ± 0.001 bc
	BEJ1	4.22 ± 0.69 ab	384.51 ± 46.54 c	375.92 ± 57.74 c	0.997 ± 0.000 bc
	BEB2	3.48 ± 0.42 ab	572.71 ± 19.17 b	668.60 ± 24.86 a	0.992 ± 0.001 d
	BEJ2	4.59 ± 0.30 a	633.01 ± 33.59 ab	644.18 ± 56.44 ab	0.992 ± 0.001 d
	BEB3	4.37 ± 0.77 ab	728.86 ± 55.29 a	724.05 ± 42.94 a	0.992 ± 0.001 d
	BEJ3	4.69 ± 0.26 a	521.60 ± 52.80 b	523.98 ± 32.61 b	0.995 ± 0.001 c
注：表中同列数据后相同字母表示采用DPS中LSD法检验差异不显著 (P>0.05)。Note: Date followed by the same letter in the same column are not significantly different by the least significant difference test (LSD) in DPS (P>0.05).

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