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云南‘兔眼’蓝莓叶内生真菌多样性分析
Diversity Analysis of Endophytic Fungi in 'Rabbit-eye' Blueberry Leaves from Yunnan
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摘要: 为探究云南栽培蓝莓叶内生真菌群落组成与多样性及其影响因素,应用Illumina高通量测序方法分析了云南省兰坪、大理以及景东产区“兔眼”蓝莓叶内生真菌多样性与群落组成差异,探究了生态气候因子与栽培管理方式对蓝莓叶内生真菌群落结构的影响,基于FUNGuild(真菌功能数据库)预测其生态功能类群。结果表明:所有样本共检测得到663个OTU,归属于8个门、31个纲、89个目、187个科、289个属和389个物种,共有OTU数量为13个,独有OTU数量依次为大理 > 兰坪 > 景东(P < 0.05);蓝莓叶内生真菌群落Alpha多样性与丰富度以大理产区最高,景东产区最低(P < 0.05);在门水平上,大理和兰坪产区以子囊菌门为主,相对丰度分别为82.13%和88.17%,景东产区优势菌门为未分类真菌(40.92%);在属水平上,兰坪产区优势属为枝孢菌属(38.99%)和(13.02%),大理产区优势属为附球菌属(20.81%)和枝孢菌属(9.66%),景东产区则以未分类真菌和未知真菌为主,占其菌群的60.82%,不同产区内生真菌优势属(排名前30)相对丰度之间存在显著差异(P < 0.05);FUNGuild真菌群落功能预测显示,病原-共生-腐生营养型在兰坪产区蓝莓叶内生真菌中占比较高,为44.65%;景东产区以未知营养型(61.6%)为主,大理产区以病原-腐生营养型(34.74%)为主;年均温和地膜覆盖栽培模式与毛孢子菌属等5个菌属相对丰度呈正相关(P < 0.05),而与枝孢菌属等6个菌属相对丰度呈负相关(P < 0.05),以上两类环境因子对优势属相对丰度的影响与其它环境因子呈相反趋势,红酵母菌属等9个菌群相对丰度不受生态气候因子与栽培管理措施的影响,可能具有更广泛的生存适应性。综上,‘兔眼’蓝莓叶内生真菌群落组成、多样性以及生态功能在云南不同产区之间存在显著差异,生态气候因子与栽培管理方式对蓝莓叶内生真菌群落组成的影响方式各有不同。Abstract: This study investigated the community composition and diversity of endophytic fungi in the leaves of "Rabbit-eye" blueberries from Lanping, Dali, and Jingdong in Yunnan Province through Illumina high-throughput sequencing. It also explored the effects of ecological and climatic factors as well as cultivation management practices on the community structure of these fungi and predicted their potential ecological functions based on the FUNGuild database. Results showed that a total of 663 OTUs were detected across all samples, classified into 8 phyla, 31 classes, 89 orders, 187 families, 289 genera, and 389 species. 13 OTUs were shared among all regions, with the number of unique OTUs following the order: Dali > Lanping > Jingdong (P < 0.05). The alpha diversity and richness of the endophytic fungal communities were the highest in Dali and lowest in Jingdong (P < 0.05). At the phylum level, Ascomycota was dominated in Dali (82.13%) and Lanping (88.17%), while the dominant phylum in Jingdong was unclassified fungi (Fungi_phy_Incertae_sedis) (40.92%). At the genus level, Cladosporium (38.99%) and Leptosphaerulina (13.02%) were the dominant genera in Lanping, Epicoccum (20.81%) and Cladosporium (9.66%) in Dali, and unclassified fungi and unknown fungi (unclassified_k__Fungi) accounted for 60.82% of the community in Jingdong. Significant differences in the relative abundance of the top 30 genera were observed among the regions (P < 0.05). Functional prediction using the FUNGuild database revealed that the Pathotroph-Saprotroph-Symbiotroph functional group was predominant in Lanping (44.65%), the unknown nutritional group in Jingdong (61.6%), and the Pathotroph-Saprotroph group in Dali (34.74%). Correlation analysis showed that annual average temperature and plastic film covering cultivation had opposite effects on the relative abundance of dominant genera compared to other environmental factors. These two factors were positively correlated with the relative abundance of five genera, including Trichosporon (P < 0.001), and negatively correlated with six genera, including Cladosporium (P < 0.05). In contrast, 9 genera, including Rhodotorula, were not significantly affected by ecological and climatic factors or cultivation practices, suggesting broader survival adaptability. In conclusion, significant differences in the community composition, diversity, and ecological functions of endophytic fungi in "Rabbit-eye" blueberry leaves were observed across different regions in Yunnan. Ecological and climatic factors and cultivation management practices had distinct impacts on the community structure of these fungi.