Diversity and community composition of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus in Beijing, northern China

doi:10.17520/biods.2023498

Abstract

Abstract:

Aims Phyllosphere fungi play crucial roles in influencing plant health and contributing to biogeochemical cycling. This study aims to investigate the diversity and community structure of epiphytic fungi in the phyllosphere of two most common evergreen trees in Beijing, namely Pinus tabuliformis and Euonymus japonicus to provide the basic data for management of urban evergreen trees.

Methods In this study, we collected leaves samples from four sites in Beijing (Andingmen area, Hepingjie area, Yayuncun area, and Beiqijia area) during four seasons (January, April, July and November) and analyzed the diversity and community composition of epiphytic fungi using high-throughput sequencing technique.

Results A total of 2,051 amplicon sequence variants (ASVs) was obtained from epiphytic fungi on P. tabuliformis while 1,970 ASVs were obtained from those on E. japonicus. The epiphytic fungi were dominated by Dothideomycetes, Leotiomycetes, and Tremellomycetes. Alpha diversity indices (Shannon diversity index, Chao1 richness index, Pielou evenness index) for epiphytic fungi varied among sampling plots but consistently showed lowest values at Hepingjie. The diversity showed a decreasing and then increasing trend from the center to north of the city. The three alpha diversity indices of fungi in P. tabuliformis varied in different months, while November had the highest index of fungi in E. japonicus during the four seasons. Result of non-metric multidimensional scaling revealed significant differences between composition patterns of epiphytic fungal communities associated with these two species (ANOSIM, P= 0.001). Linear discriminant analysis effect size identified significant biomarkers specific to each studied species, seven genera (e.g., Aureobasidium, Paraconiothyrium, Orbilia) served as biomarkers for P. tabuliformis, while seven fungal genera (e.g., Erysiphe, Alternaria, Nigrospora) represented biomarkers for E. japonicus. Regression analyses showed that mean precipitation was significantly and positively correlated with the abundance of epiphytic fungi in P. tabuliformis, and significantly and negatively correlated with the abundance of epiphytic fungi in E. japonicus. Network analysis identified the dominant core fungal genera for E. japonicus are Vacuiphoma, Mrakia, Curvularia, and Erysiphe, whereas Vacuiphoma was single core fungal genus for P. tabuliformis. The main functional groups of epiphytic fungi in the phyllosphere of P. tabuliformis and E. japonicus were pathotrophic and saprotrophic, respectively.

Conclusion The results of this study indicate that there is a significant difference in the composition of the epiphytic fungi in the phyllosphere between the two evergreen trees species and there are some patterns in the α diversity changes of different sites and seasons. Plant leaf diseases are most severe in April and should be emphasized for prevention and control.

Key words: epiphytic fungi in the phyllosphere, urban greening trees, Illumina MiSeq sequencing, fungal diversity, core fungal communities, network analysis

Yujin Cui, Wanying Li, Qingqing Zhou, Heng Zhao, Fang Wu, Yuan Yuan. Diversity and community composition of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus in Beijing, northern China[J]. Biodiv Sci, 2024, 32(7): 23498.

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URL: https://www.biodiversity-science.net/EN/10.17520/biods.2023498

https://www.biodiversity-science.net/EN/Y2024/V32/I7/23498

Figures/Tables 14

Fig. 1 The sampling sites of Pinus tabuliformis and Euonymus japonicus samples. ADM, Andingmen; HPJ, Hepingjie; YYC, Yayuncun; BQJ, Beiqijia.

Fig. 2 Venn diagram of distribution of ASVs for different groups of fungi. A, Pinus tabuliformis in different sampling plots; B, P. tabuliformis in different seasons; C, Euonymus japonicus in different sampling plots; D, E. japonicus in different seasons; E, Between the two trees species.

Table 1 The alpha diversity index of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus among different sampling plots

树种 Tree species	样点 Sample plot	Shannon 多样性指数 Shannon diversity index	Chao1丰富度指数 Chao1 richness index	Pielou均匀度指数 Pielou evenness index
油松 Pinus tabuliformis	安定门 ADM	3.92 ± 0.64^a	193.63 ± 83.48^a	0.53 ± 0.06^a
	和平街 HPJ	3.57 ± 1.06^a	161.04 ± 74.94^a	0.50 ± 0.13^a
	亚运村 YYC	3.87 ± 0.43^a	171.96 ± 60.35^a	0.52 ± 0.05^a
	北七家 BQJ	4.03 ± 0.42^a	174.95 ± 56.75^a	0.55 ± 0.07^a
大叶黄杨Euonymus japonicus	安定门 ADM	3.47 ± 0.62^a	190.72 ± 58.41^a	0.46 ± 0.07^a
	和平街 HPJ	2.69 ± 0.37^b	129.80 ± 39.43^b	0.39 ± 0.046^b
	亚运村 YYC	2.77 ± 0.98^b	143.34 ± 68.27^b	0.40 ± 0.13^b
	北七家 BQJ	3.06 ± 0.66^ab	192.09 ± 77.21^a	0.41 ± 0.06^b

Fig. 3 The diversity and class level community structure of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus among different sampling plots. A, The alpha diversity index of P. tabuliformis; B, The alpha diversity index of E. japonicus; C, Non-metric multidimensional scaling analysis of P. tabuliformis; D, Non-metric multidimensional scaling analysis of E. japonicus; E, The community structure of P. tabuliformis; F, The community structure of E. japonicus. Different letters in the box plot indicates that there was significant difference between groups (Kruskal-Wallis, P < 0.05). r denotes the degree of similarity between sample points. The larger the value of, the greater the difference. Full names of sample plot abbreviations see Fig. 1.

Table 2 The analysis of similarities of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus among different sampling plots

树种 Tree species	样点间对比 Comparison between sample plots	r	P
油松 Pinus tabuliformis	安定门/和平街 ADM/HPJ	0.4016	0.001
	安定门/亚运村 ADM/YYC	0.2931	0.001
	安定门/北七家 ADM/BQJ	0.4734	0.001
	和平街/亚运村 HPJ/YYC	0.0273	0.133
	和平街/北七家 HPJ/BQJ	0.2422	0.001
	亚运村/北七家 YYC/BQJ	0.2056	0.001
大叶黄杨Euonymus japonicus	亚运村/北七家 YYC/BQJ	0.1251	0.006
	亚运村/安定门 YYC/ADM	0.2292	0.001
	亚运村/和平街 YYC/HPJ	0.0683	0.044
	北七家/安定门 BQJ/ADM	0.2782	0.001
	北七家/和平街 BQJ/HPJ	0.1253	0.006
	安定门/和平街 ADM/HPJ	0.3554	0.001

Table 3 The alpha diversity index of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus among different seasons

树种 Tree Species	季节 Seasons	Shannon多样性指数Shannon diversity index	Chao1丰富度指数 Chao1 richness index	Pielou均匀度指数Pielou evenness index
油松 Pinus tabuliformis	七月 July	3.96 ± 0.34^a	206.10 ± 31.82^a	0.52 ± 0.037^a
	十一月 Nov.	3.70 ± 1.13^a	149.69 ± 102.57^c	0.53 ± 0.14^a
	一月 Jan.	3.93 ± 0.63^a	174.34 ± 73.91^abc	0.54 ± 0.06^a
	四月 Apr.	3.79 ± 0.35^a	171.99 ± 33.95^b	0.51 ± 0.04^a
大叶黄杨Euonymus japonicus	七月 July	2.78 ± 0.26^bc	118.78 ± 34.40^b	0.41 ± 0.04^b
	十一月 Nov.	3.47 ± 0.60^a	185.69 ± 76.38^a	0.47 ± 0.06^a
	一月 Jan.	2.74 ± 0.58^c	179.19 ± 71.27^a	0.37 ± 0.05^c
	四月 Apr.	3.00 ± 1.11^ab	174.52 ± 61.58^a	0.40 ± 0.14^ab

Fig. 4 The diversity and class level community structure of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus across different seasons. A, The alpha diversity index of P. tabuliformis; B, The alpha diversity index of E. japonicus; C, Non-metric multidimensional scaling analysis of P. tabuliformis; D, Non-metric multidimensional scaling analysis of E. japonicus; E, The community structure of P. tabuliformis; F, The community structure of E. japonicus. Different letters in the box plot indicates that there was significant difference between groups (Kruskal-Wallis, P < 0.05). The larger the value of, the greater the difference.

Table 4 The alpha diversity index of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus among different seasons

	季节间对比 Comparison between seasons	r	P
油松 Pinus tabuliformis	七月/十一月 July/Nov.	0.1533	0.001
	七月/一月 July/Jan.	0.1938	0.001
	七月/四月 July/Apr.	0.2540	0.001
	十一月/一月 Nov./Jan.	0.0235	0.185
	十一月/四月 Nov./Apr.	0.1882	0.002
	一月/四月 Jan./Apr.	0.2520	0.001
大叶黄杨 Euonymus japonicus	十一月/七月 Nov./July	0.1918	0.001
	十一月/一月 Nov./Jan.	0.2062	0.002
	十一月/四月 Nov./Apr.	0.8448	0.001
	七月/一月 July/Jan.	0.1439	0.009
	七月/四月 July/Apr.	0.8940	0.001
	一月/四月 Jan./Apr.	0.9305	0.001

Fig. 5 The diversity and community structure of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus at different seasons; A, Heat map of top 30 genera in relative abundance ranking of fungi (data processed by log10); B, Sankey diagram of two trees; C, Non-metric multidimensional scaling analysis of two trees; D, Linear discriminant analysis effect size (LEfSe) of two trees; E, The alpha diversity index of two trees. YS, P. tabuliformis, HY, E. japonicus. The “*” in heat map and different letters in the box plot indicates that there was significant difference between groups (Kruskal-Wallis, P < 0.05).

Fig. 6 Variance partitioning analysis of community composition and temperature, PM2.5, and precipitation of Euonymus japonicas (A) and Pinus tabuliformis (B). Values < 0 was not shown.

Fig. 7 Linear regression analysis of Chao1 richness index of epiphytic fungi in the phyllosphere with temperature, PM2.5 and precipitation in Pinus tabuliformis (A) and Euonymus japonicus (B)

Fig. 8 Network of the top 40 epiphytic fungal genera in relative abundance in the phyllosphere of Pinus tabuliformis (A) and Euonymus japonicus (B). The thickness of the line represents the strength of the correlation relationship and the size of the circle represents the size of the degree.

Table 5 Functional classification of epiphytic fungi in phyllosphere of Pinus tabuliformis and Euonymus japonicus

营养类型 Trophic mode	功能群 Guild	油松功能群比例 Proportion of guild in Pinus tabuliformis (%)	营养类型占比 Proportion of trophic mode (%)	大叶黄杨功能群比例 Proportion of guild in Euonymus japonicus (%)	营养类型占比Proportion of trophic mode (%)
腐生营养型 Saprotroph	未定义腐生真菌 Undefined saprotroph fungi	46.824	63.619	21.269	22.215
	粪腐生真菌 Dung saprotroph fungi	0.106		0.061
	木腐生真菌 Wood saprotroph fungi	16.668		0.630
	土壤腐生真菌 Soil saprotroph fungi	0.018		0.240
	叶片腐生真菌 Leaf saprotroph fungi	0.001		0.015
	凋落物腐生真菌 Litter saprotroph fungi	0.003		0.001
共生营养型 Symbiotroph	内生真菌 Endophyte fungi	0.595	0.651	0.266	0.315
	外生菌根真菌 Ectomycorrhizal fungi	0.005		0
	杜鹃类菌根真菌 Ericoid mycorrhizal fungi	0.002		0.001
	植物附生真菌 Epiphyte fungi	0.049		0.048
病理营养型 Pathotroph	植物病原真菌 Plant pathogen fungi	9.171	9.246	70.806	70.906
	动物病原真菌 Animal pathogen fungi	0.023		0.025
	真菌寄生真菌 Fungal parasite fungi	0.037		0.070
	苔藓植物寄生真菌 Bryophyte parasite fungi	0.002		0.001
	地衣寄生菌 Lichen parasite fungi	0.013		0.004
	其他 Other	26.484		6.564

Fig. 9 Trophic mode of epiphytic fungi in the phyllosphere of Pinus tabuliformis and Euonymus japonicus. A, P. tabuliformis in different sampling plots; B, P. tabuliformis in different seasons; C, E. japonicus in different sampling plots; D, E. japonicus in different seasons. Full names of sample plot abbreviations see Fig. 1.

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