感赤星病与健康烟叶在腈菌唑应用后的叶际微生态

doi:10.3969/j.issn.1004-1524.20230242

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (1): 156-167.DOI: 10.3969/j.issn.1004-1524.20230242

感赤星病与健康烟叶在腈菌唑应用后的叶际微生态

张艺¹^,²(), 王丰²^,^*(), 蔡刘体², 汪汉成², 熊晶³, 陈兴江²

1.贵州大学农学院,贵州贵阳 550025
2.贵州省烟草科学研究院,贵州贵阳 550081
3.贵州省烟草公司毕节市公司,贵州毕节 551700

收稿日期:2023-02-28 出版日期:2024-01-25 发布日期:2024-02-18
作者简介:张艺(1996—),女,贵州遵义人,硕士研究生,研究方向为作物栽培学与耕作学。E-mail:1158852415@qq.com
通讯作者: * 王丰, E-mail:yancaowangfeng@163.com
基金资助:
贵州省“百层次”创新型人才(黔科合平台人才-GCC〔2022〕028-1);国家自然科学基金(31960550);国家自然科学基金(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));中国烟草总公司科技项目(110202101048(LS-08))

Phyllosphere microecology of brown spot and healthy tobacco leaves after application of myclobutanil

ZHANG Yi¹^,²(), WANG Feng²^,^*(), CAI Liuti², WANG Hancheng², XIONG Jing³, CHEN Xingjiang²

1. College of Agriculture, Guizhou University, Guiyang 550025,China
2. Guizhou Academy of Tobacco Science, Guiyang 550081,China
3. Bijie Tobacco Company of Guizhou Province, Bijie 551700, Guizhou, China

Received:2023-02-28 Online:2024-01-25 Published:2024-02-18

摘要/Abstract

摘要：

为了解腈菌唑在防控赤星病时,烟叶叶际微生物菌群结构与代谢功能的差异,采用Illumina高通量测序技术分析了感病与健康烟叶真菌和细菌的菌群结构与多样性特征,采用Biolog-ECO技术分析其代谢功能。结果表明,药前感病烟叶与健康烟叶叶际真菌有链格孢属、谷菌根菌属、Boeremia、枝孢霉属等,其中链格孢属为优势菌属;叶际细菌有科萨克氏菌属、泛菌属、甲基杆菌属等,其中科萨克氏菌属为优势菌属;感病烟叶链格孢属和科萨克氏菌属相对丰度均高于健康烟叶。每667 m²采用15%腈菌唑微乳剂60 mL处理5、10、15 d时,感病烟叶叶际真菌链格孢属和枝孢霉属相对丰度均降低,附球菌属和亚隔孢壳属相对丰度均先降低后增加;健康烟叶叶际真菌链格孢属、Symmetrospora、亚隔孢壳属的相对丰度均下降。感病烟叶叶际细菌科萨克氏菌属相对丰度在施药5 d时降低,10 d和15 d时增加,假单胞菌属与之相反;健康烟叶叶际细菌泛菌属、假单胞菌属相对丰度在施药5 d和10 d时降低,15 d时增加。感病烟叶叶际微生物代谢活性弱于健康烟叶,叶际微生物对除α-丁酮酸、L-苏氨酸和4-羟基苯甲酸外的碳源代谢均较强。腈菌唑应用后,感病与健康烟叶叶际微生物的代谢活性均下降,其代谢活性均随施药时间的延长而逐渐恢复。研究结果揭示了腈菌唑施用不同时期感病与健康烟叶组织叶际微生态变化差异,为腈菌唑防控烟草赤星病的科学应用提供了参考依据。

关键词: 烟草赤星病, 腈菌唑, 叶际微生态, 链格孢属, 代谢功能

Abstract:

To understand the differences in the structure and metabolic functions of the phyllosphere microorganisms of tobacco leaves in the control of brown spot by myclobutanil, Illumina high-throughput sequencing and Biolog-ECO technology was conducted to investigate the structure and diversity of phyllospheric microorganisms community and their metabolic function. The results showed that the phyllosphere fungi of the diseased and healthy tobacco leaves included Alternaria, Archaeorhizomyces, Boeremia, Cladosporium, etc., of which Alternaria was the dominant genus; the bacteria were Kosakonia, Pantoea and Methylobacterium. Among them, Kosakonia is the dominant genus, and the relative abundance of the Alternaria and Kosakonia was higher in diseased tobacco leaves than in healthy tobacco leaves. The relative abundance of Alternaria and Cladosporium decreased, while the relative abundance of Epicoccum and Didymella decreased and then increased when treated with 60 mL 15% myclobutanil microemulsion per 667 m² for 5, 10 and 15 d; the relative abundance of phyllosphere fungi in healthy tobacco leaves decreased in the genera Alternaria, Symmetrospora, and Didymella. The relative abundance of phyllosphere Kosakonia in diseased tobacco leaves decreased at 5 d of application and increased at 10 and 15 d, while the opposite was true for Pseudomonas. The relative abundance of Pantoea and Pseudomonas decreased at 5 and 10 d and increased at 15 d. The phyllospheric microorganism metabolic activity of brown spot-infected leaves was lower than that of healthy tobacco leaves. The metabolism of carbon sources other than α-butyric acid, L-threonine and 4-hydroxybenzoic acid by phyllosphere microorganisms was strong. The metabolic activity of phyllosphere microorganisms in diseased and healthy tobacco leaves were reduced after myclobutanil treatment, and the metabolic activity of phyllospheric microorganisms gradually recovered with the increasing of application time. The results of the study revealed the differences in phyllosphere microecological changes between diseased and healthy tobacco leaves during different periods of myclobutanil application, providing a reference for the scientific application of myclobutanil in the control of tobacco brown spot.

Key words: tobacco brown spot, myclobutanil, phyllosphere microecology, Alternaria sp., metabolic function

中图分类号:

S482.2

张艺, 王丰, 蔡刘体, 汪汉成, 熊晶, 陈兴江. 感赤星病与健康烟叶在腈菌唑应用后的叶际微生态[J]. 浙江农业学报, 2024, 36(1): 156-167.

ZHANG Yi, WANG Feng, CAI Liuti, WANG Hancheng, XIONG Jing, CHEN Xingjiang. Phyllosphere microecology of brown spot and healthy tobacco leaves after application of myclobutanil[J]. Acta Agriculturae Zhejiangensis, 2024, 36(1): 156-167.

图/表 7

表1 样品采集信息

Table 1 Samples information

样品采集 Sampling		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.	JJJ01	JJJ11	JJJ21	JJJ31
		JJJ02	JJJ12	JJJ22	JJJ32
		JJJ03	JJJ13	JJJ23	JJJ33
	分组编号Group No.	JJJ0	JJJ1	JJJ2	JJJ3
感病组织Diseased tissue	样品编号Sample No.	JJB01	JJB11	JJB21	JJB31
		JJB02	JJB12	JJB22	JJB32
		JJB03	JJB13	JJB23	JJB33
	分组编号Group No.	JJB0	JJB1	JJB2	JJB3

图1 烟叶样品真菌、细菌的OTU测序深度和水平物种累积箱形图 A,真菌OTU测序深度;B,细菌OTU测序深度;C,真菌水平物种累积箱形图;D细菌水平物种累积箱形图。

Fig.1 Box diagram of OTU sequencing depth and horizontal species accumulation for fungi and bacteria in tobacco samples A, OTU sequencing depth of fungi; B, OTU sequencing depth of bacteria; C, Horizontal species accumulation of fungi; D, Horizontal species accumulation of bacteria.

表2 烟叶叶际真菌和细菌Alpha多样性差异(OTU水平)

Table 2 Alpha diversity of fungi and bacteria in leaf interleaf (OTU level)

类型 Type	样品 Sample	多样性指数Diversity index	丰富度指数Richness index	覆盖度指数 Coverage index
类型 Type	样品 Sample	Shannon	ACE	覆盖度指数 Coverage index
真菌Fungi	JJB0	0.99±0.72 d	40.08±18.27 e	0.99±0.00 a
	JJJ0	3.08±0.23 bc	111.36±30.10 cd	0.99±0.00 b
	JJB1	1.24±1.23 d	45.89±22.40 e	0.99±0.00 ab
	JJJ1	3.53±0.28 b	130.05±47.47 c	0.99±0.01 b
	JJB2	0.75±0.32 ad	62.88±40.26 de	0.99±0.00 ab
	JJJ2	4.79±0.22 a	242.72±28.89 b	0.98±0.01 c
	JJB3	2.48±0.39 c	254.21±6.89 b	0.97±0.00 c
	JJJ3	4.84±0.14 a	323.81±26.59 b	0.97±0.00 d
细菌Bacteria	JJB0	1.25±0.95 c	32.09±9.86 b	0.98±0.01 ab
	JJJ0	1.19±0.06 c	22.12±6.16 b	0.98±0.01 a
	JJB1	1.21±0.71 c	17.02±5.36 b	0.99±0.00 a
	JJJ1	0.66±0.28 c	28.47±14.15 b	0.97±0.01 ab
	JJB2	2.58±0.78 ab	264.20±299.63 ab	0.87±0.10 bc
	JJJ2	1.49±0.26 bc	328.64±192.21 ab	0.88±0.03 abc
	JJB3	3.34±1.36 a	623.03±524.08 a	0.81±0.11 c
	JJJ3	2.94±0.49 a	482.17±272.18 a	0.83±0.07 c

图2 感病与健康烟叶真菌和细菌群落组成相对丰度图 A、C分别为真菌门水平和属水平;B、D分别为细菌门水平和属水平。

Fig.2 Relative abundance of fungal and bacterial community composition of diseased and healthy tobacco leaves A and C are at the fungal phylum and genus level, respectively; B and D are at the bacterial phylum and genus level, respectively.

表3 环境因子与病情指数

Table 3 Environmental factors and disease index

环境因子及病情指数	施药前0 d	施药后5 d	施药后10 d	施药后15 d
Environmental factors and disease index	0 day before application	5 days after application	10 days after application	15 days after application
降雨量Rainfall/mm	13.60	19.00	11.30	25.40
气温Temperature/℃	18.32	18.97	17.51	16.54
空气相对湿度Relative humidity/%	82.49	83.81	78.99	90.98
病情指数Disease index	44.44	48.14	63.58	70.55

图3 腈菌唑处理后叶际真菌(A)和细菌(B)群落的Spearman相关分析热图

Fig.3 Heat map of Spearman correlation analysis based on phyllosphere fungal (A) and bacterial (B) communities after myclobutanil treatment *, P<0.05; **, P<0.01.

图4 烟叶叶际微生物代谢功能聚类热图颜色值分别代表微生物代谢碳源的程度,122~<192为代谢程度低,192~262为代谢程度一般,262以上为代谢程度高。

Fig.4 Cluster heat map of phyllosphere microbial metabolic function in tobacco leaves The color values represent the degree of the microbes metabolizing carbon sources. The value 122-<192, 192-262 and >262 represent the leaf microbes metabolized in Biolog ECO microplate poorly, moderately, effectively, respectively.

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感赤星病与健康烟叶在腈菌唑应用后的叶际微生态

Phyllosphere microecology of brown spot and healthy tobacco leaves after application of myclobutanil

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