Background: The difference of metabolites in medicinal plants has always been concerned to be influenced by external environmental factors. However, the relationship between endophytes and host metabolites remains unclear. Methods: In this study, we used 16S and ITS amplicon sequencing to compare endophyte diversity among different tissue types and ages of Gentiana officinalis. Endophyte diversity and abundance was also analyzed in relation to the abundance of four secondary metabolites (Gentiopicroside, Loganic acid, Swertiamarine and Sweroside). Results: The diversity and richness of G. officinalis endophyte differed as a function of tissue types and ages. Four metabolites of G. officinalis were significantly correlated with the abundance of dominant endophyte genera. The predictive function analysis showed that metabolism was main function of endophytic bacteria in different tissue and year root samples, while saprotroph was dominant trophic modes of endophytic fungi in the different year root samples. The dominant trophic modes of endophytic fungi was saprotroph and pathotroph, and relative abundances differed in the different tissue samples. The results of this study will help to elucidate the plant-microbial interactions and provide key information on the role of endophytes in the production of G.officinalis and its important metabolites.

The lack of diagnostic taxonomic characteristics in some species complexes leave the species delimitation of Oxytropis DC. unresolved. Seed morphological features have proved to be useful diagnostic and taxonomic characteristics in Fabaceae. However, there are few systematic studies on the seed characteristics of Oxytropis. Here, we used scanning electron and stereoscopic microscopy to investigate the seed characteristics of 35 samples obtained from 21 Oxytropis species from northwest China. Our examination showed two main types of hilum positions, terminal and central, and five different types of seed shapes: prolonged semielliptic, reniform, prolonged reniform, quadratic, and cardiform. Seven different sculpturing patterns were identified: scaled, regulated, lophate with stellated testa cells, simple reticulate, rough, compound reticulate, and lophate with rounded testa cells. The seeds ranged from 1.27 to 2.57 mm in length and from 1.18 to 2.02 mm in width, and the length-to-width ratio ranged from 0.89 to 1.55 mm. The seed shape was constant within species and was useful for species delimitation within the genus Oxytropis when combined with other macroscopic traits. In contrast, the sculpturing patterns were highly variable at the species level and could not be used for species identification. Results of the cluster analysis and principal component analysis (PCA) indicated that the seed traits of Oxytropis species are useful for taxa identification at the species level, but have low taxonomic value at the section level.

Repeatable floral closure with diurnal rhythms, that is, flower opening in the morning and closing in the evening, was widely reported. However, the rhythm of flower opening in the morning but closing in the midday received much less attention. Gentianopsis paludosa, Gentianaceae, has an obvious petal movement rhythm opening in the morning and closing at noon at northeast of the Qinghai‐Tibetan Plateau. In this study, we examined the effects of temperature (T), relative humidity (RH), and illumination intensity (II) on G. paludosa's flower closure. Furthermore, we monitored the environmental changes inside and outside of the flowers, aiming to test the effect of floral closure on the stability of microenvironment inside the flower. Finally, we artificially interrupted temporal petal closure and investigated its effects on reproductive fitness. The results showed that high/low temperature contributed more to the flower closure than low RH, while illumination intensity had no significant effect on it. The medium temperature, relative humidity and illumination intensity (environmental conditions at 10:00) did not delay flower closure when flowers at pre‐closing period or stimulate reopen when flowers full closed. Floral closure provided a stable temperature condition and a higher RH condition inside the flower. Meanwhile, compulsive opening and delayed closure of flowers decreased the seed‐set ratio while no effect was found when flowers were forced to close. We conclude that endogenous rhythm regulates floral closure. The rhythm of petal movement providing a stable microenvironment for G. paludosa, increasing the seed production and saving energy from flower opening maintenance, which might be an adaptive strategy to against unfavorable environmental conditions.

Soil heavy metal contamination has become a global environmental issue, which threaten soil quality, food security and human health. Symphytum officinale L. have exhibited high tolerance and restoration capacity to heavy metals (HMs) stress. However, little is known about the mechanisms of HMs in S. officinale. In this study, transcriptomic and physiological changes of S. officinale response to different HMs (Pb, Cd and Zn) were analyzed and investigated the key genes and pathways involved in HMs uptake patterns. The results showed that phenotypic effects are not significant, and antioxidant enzyme activities were all upregulated. Transcriptome analysis indicated that 1247 differential genes were up-regulated, and 1963 differential genes were down-regulated under Cd stress, while 3752 differential genes were up-regulated, and 7197 differential genes were down-regulated under Pb stress; and 527 differential genes were up-regulated; and 722 differential genes were down-regulated under Zn stress. Based on their expression, we preliminarily speculate that different HMs resistance of S. officinale may be regulated by the differential expression of key genes. These results provide a theoretical basis for determining the exact expression of genes in plants under different heavy metal stress, the processes involved molecular pathways, and how they can be efficiently utilized to improve plant tolerance to toxic metals and improve phytoremediation efficiency.

During the evolution of angiosperm flowers, some floral traits may undergo certain changes in order to participate in screening. The stamens and pistils of Delphinium caeruleum are covered by two “door-like” staminodes, the evolutionary function of which, however, is quite unknown. In this study, we investigated whether D. caeruleum staminodes acted as visitor filters by assessing the respective strengths of staminodes and visitor insects (six bee species). We measured the operative strength required to open the staminodes and the strength that insects were capable of exerting using a biological tension sensor. Furthermore, we compared the strength required to open staminodes at different phases of the flowering period (male and female phases) and the strength of different visitors (visitors and non-visitors of D. caeruleum). The results showed that the strength needed to open staminodes in the male phase was significantly higher than that in the female phase. There was no significant difference between the strength exerted by visitors and required by staminodes of D. caeruleum in the male phase, but the visitor strength was significantly higher than that required to open staminodes in the female phase flowers. The strength of non-visitors was significantly lower than that required to open staminodes in the male phase. Furthermore, there was a significant positive association between the strength and the body weight of the bees. These results highlighted the observation that only strong visitors could press the two staminodes to access the sex organs and achieve successful pollination. Furthermore, these results revealed the function of pollinator screening by the staminodes of D. caeruleum. The biomechanical approach to the study of flowers allowed us to address relevant ecological and evolutionary questions of the plant–pollinator interaction and explore the functional modules within the flower structure in other plant species.

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.