Endophytes play an important role in helping plants resist heavy metal stress. However, little is known about the effects of different heavy metals on the diversity and composition of endophyte communities. In this study, we used 16S and ITS amplicon sequencing to reveal the structure and function of endophytes in Symphytum officinale under different heavy metal stressors. The results showed that the endophytic fungal diversity decreased compared with the control under the different heavy metals stressors, while the diversity of endophytic bacteria showed an increasing trend. The biomarker analysis indicated that Zn and Pb stress led to obvious branches. Specific OTUs analysis showed that there were 1224, 597, and 1004 OTUs specific under Zn, Pb, and Cd stress in the bacterial community and 135, 81, and 110 OTUs specific under Zn, Pb, and Cd stress in the fungal community. The co-occurrence network showed changes in microbial interactions under heavy metal contamination conditions, suggesting that endophytic bacteria play an important role in the resistance of host plants. The Spearman analysis showed that the correlation between endophytic bacteria and endophytic fungi in relation to heavy metal transport exhibited variations. Our results expand the knowledge of the relationships of plant–microbe interactions and offer pivotal information to reveal the role of endophytes under different heavy metal stress conditions.
Nectar robbers, which affect plant fitness (directly or indirectly) in different degrees and in different ways, potentially constitute a significant part of mutualistic relationships. While the negative effects of nectar robbing on plant reproductive success have been widely reported, the positive effects remain unknown. The target of our study was to evaluate the effects of nectar robbers on the reproductive success of Symphytum officinale (Boraginaceae). We observed the behavior, species and times of visitors in the field, and we assessed the effect of nectar robbers on corolla abscission rate and time. To test the fitness of corolla abscission, we detected the changes in stigma receptivity, pollen viability, pollen amount and appendage opening size along with the time of flower blossom. The flowering dynamics and floral structure were observed to reveal the mechanism of self-pollination. Finally, pollen deposition seed set rate and fruit set rate were determined to estimate the effect of nectar robbers on reproduction success. We observed 14 species of visitors and 2539 visits in 50 h of observation; 91.7% of them were nectar robbers. The pressure and nectar removal of nectar robbers significantly promoted corolla abscission during a period when pollen grains are viable and the stigma is receptive. In addition, corolla abscission significantly increased the pollen deposition and seed setting rate. Our results demonstrate that nectar robbing contributes to enhancing seed production and positively and indirectly impacts the reproductive success of S. officinale. This mechanism involved the movement of anthers and indirect participation by nectar robbers, which was rarely investigated. Considering the multiple consequences of nectar robbing, understanding the impact of nectar robbers on plant reproduction is essential to comprehend the evolutionary importance of relationships between plants and their visitors.