Nectar robbing by bees affects the reproductive fitness of the distylous plant Tirpitzia sinensis (Linaceae).

Nectar robbing can affect plant reproductive success directly by influencing female and male fitness, and indirectly by affecting pollinator behavior. Flowers have morphological and chemical features that may protect them from nectar robbers. Previous studies on nectar robbing have focused mainly on homotypic plants. It remains unclear how nectar robbing affects the reproductive success of distylous plants, and whether defense strategies of two morphs are different. Nectar-robbing rates on the long- and short-styled morph (L-morph, S-morph) of the distylous Tirpitzia sinensis were investigated. We compared floral traits, the temporal pattern of change in nectar volume and sugar concentration, nectar secondary metabolites, and sugar composition between robbed and unrobbed flowers of two morphs. We tested direct effects of nectar robbing on female and male components of plant fitness and indirect effects of nectar robbing via pollinators. Nectar-robbing rates did not differ between the two morphs. Flowers with smaller sepals and petals were more easily robbed. The floral tube diameter and thickness were greater in L-morphs than in S-morphs, and the nectar rob holes were significantly smaller in L-morphs than in S-morphs. Nectar robbing significantly decreased nectar replenishment rate but did not affect nectar sugar concentration or sugar composition. After robbery, the quantities and diversity of secondary compounds in the nectar of S-morphs increased significantly and total relative contents of secondary compounds in L-morphs showed no obvious changes. Nectar robbing could decrease female fitness by decreasing pollen germination rate and thus decreasing seed set. Nectar robbing had no significant effects on male fitness. Robbed flowers were less likely to be visited by hawkmoth pollinators, especially in S-morphs. These results suggest that nectar robbing could directly and indirectly decrease the female fitness of T. sinensis, and different morphs have evolved different defense mechanisms in response to nectar-robbing pressure.

Stronger susceptibilities to air pollutants of influenza A than B were identified in subtropical Shenzhen, China.

Air pollution was indicated to be a key factor contributing to the aggressive spread of influenza viruses, whereas uncertainty still exists regarding to whether distinctions exist between influenza subtypes. Our study quantified the impact of five air pollutants on influenza subtype outbreaks in Shenzhen, China, a densely populated and highly urbanized megacity. Daily influenza outbreak data of laboratory-confirmed positive cases were obtained from the Shenzhen CDC, from May 1, 2013 to Dec 31, 2015. Concentrations of nitrogen dioxide (NO2), sulfur dioxide (SO2), particulate matters ≤2.5 µm (PM2.5), particulate matters ≤10 µm (PM10), and ozone (O3), were retrieved from the 18 national monitoring stations. The generalized additive model (GAM) and distributed lag non-linear model (DLNM) were used to calculate the concentration-response relationships between environmental inducers and outbreak epidemics, respectively for influenza A (Flu-A) and B (Flu-B). There were 1687 positive specimens were confirmed during the study period. The cold season was restricted from Nov. 4th to Apr. 20th, covering all seasons other than the long-lasting summer. Relatively heavy fine particle matter (PM2.5) and NO2 pollution was observed in cold months, with mean concentrations of 46.06 µg/m3 and 40.03 µg/m3, respectively. Time-series analysis indicated that high concentrations of NO2, PM2.5, PM10, and O3 were associated with more influenza outbreaks at short lag periods (0-5 d). Although more Flu-B (679 cases) epidemics occurred than Flu-A (382 cases) in the cold season, Flu-A generally showed higher susceptibility to air pollutants. A 10 µg/m3 increment in concentrations of PM2.5, PM10, and O3 at lag 04, was associated with a 2.103 (95%CI: 1.528-2.893), 1.618 (95%CI: 1.311-1.996), and 1.569 (95%CI: 1.214-2.028) of the relative risk (RR) of Flu-A, respectively. A 5 µg/m3 increase in NO2 was associated with higher risk of Flu-A at lag 03 (RR = 1.646, 95%CI: 1.295-2.092) and of Flu-B at lag 04 (RR = 1.319, 95%CI: 1.095-1.588). Nevertheless, barely significant effect of particulate matters (PM2.5, PM10) on Flu-B and SO2 on both subtypes was detected. Further, the effect estimates of NO2 increased for both subtypes when coexisting with other pollutants. This study provides evidence that declining concentrations of main pollutants including NO2, O3, and particulate matters, could substantially decrease influenza risk in subtropical Shenzhen, especially for influenza A.

Ancillary polymorphic floral traits between two morphs adaptive to hawkmoth pollination in distylous plant Tirpitzia sinensis (Linaceae).

BACKGROUND: Floral morphs are characterized differentiations in reciprocal positions of sexual organs and ancillary floral traits in heterostylous plants. However, it remains unclear how differential floral morphs ensure reproductive success between morphs using the same pollinator. RESULTS: Measurements of floral traits in white-flowered Tirpitzia sinensis with long corolla tubes indicated that it is typically distylous, long-styled (L-) morph producing more but smaller pollen grains per flower than short-styled (S-) morph. Both morphs secreted more nectar volume at night than in the day and the sugar composition was rich in sucrose, potentially adaptive to pollination by hawkmoths (Macroglossum spp.) which were active at dusk. A bumblebee species functioned as the nectar robber in both morphs and a honeybee as the pollen feeder in the S-morph. The L-morph secreted more nectar volume but relatively lower sucrose/hexose ratio than the S-morph. Floral visitation rate by hawkmoths was higher but its pollination efficiency was lower in the S-morph than the L-morph. Hand pollination treatments indicated self-incompatibility in T. sinensis and seed set of open-pollinated flowers did not differ between morphs. CONCLUSIONS: Our findings suggest that the two morphs differ with respect to traits relevant to pollination. The L-morph, with its exserted stigma, has more pollen grains per anther and a greater volume of nectar, which may prolong the foraging time and increase the pollination efficiency of hawkmoths. The S-morph has a higher sucrose/hexose ratio in its nectar which can be more attractive to hawkmoths and increase the visit rates. Ancillary polymorphic floral traits between two morphs are adaptive to hawkmoth and ensure reproductive success in distylous plant T. sinensis.

The complete chloroplast genome sequence of Magnolia coriacea (Magnoliaceae), a critically endangered endemic to China.

Magnolia coriacea, Chang et B. L. (Magnoliaceae) is a critically endangered tree, endemic to Yunnan province, China. In this study, the complete chloroplast genome of M. coriacea was sequenced and analyzed. The total chloroplast genome size of M. coriacea is 160,113 bp, including a pair of inverted repeat regions (IRs, 26,576 bp) separated by a large single-copy (LSC, 88,175 bp) region and a small single-copy region (SSC, 18,786 bp). The complete chloroplast genome contains 86 protein-coding (PCGs), 37 transfer RNA (tRNAs), and 8 ribosomal RNA (rRNAs) genes. The phylogenetic analysis showed that M. coriacea is closely related to M. cathcartii. This study contributes to the bioinformatics on the evolution, genetic, conservation, and molecular biology for future studies of Magnoliaceae.

The complete chloroplast genome of Impatiens mengtszeana (Balsaminaceae), an endemic species in China.

Impatiens mengtszeana is an endemic species in China. In this study, the complete chloroplast genome of I. mengtszeana was sequenced and analyzed. The total chloroplast genome size of I. mengtszeana is 152,928 bp, including a pair of inverted repeat regions (IRs, 26,007 bp) separated by a large single copy (LSC, 83,722 bp) region and a small single copy region (SSC, 17,192 bp). The whole chloroplast genome contains 89 protein-coding genes (PCGs), 37 transfer RNA genes (tRNAs), and eight ribosomal RNA genes (rRNAs). According to the phylogenetic topologies, I. mengtszeana was closely related to I. hawkeri.

Size-dependent and environment-mediated shifts in leaf traits of a deciduous tree species in a subtropical forest.

AIMS: Understanding the joint effects of plant development and environment on shifts of intraspecific leaf traits will advance the understandings of the causes of intraspecific trait variation. We address this question by focusing on a widespread species Clausena dunniana in a subtropical broad-leaved forest. METHODS: We sampled 262 individuals of C. dunniana at two major topographic habitat types, the slope and hilltop, within the karst forests in Maolan Nature Reserve in southwestern China. We measured individual plant level leaf traits (i.e., specific leaf area (SLA), leaf area, leaf dry-matter content (LDMC), and leaf thickness) that are associated with plant resource-use strategies. We adopted a linear mixed-effects model in which the plant size (i.e., the first principal component of plant basal diameter and plant height) and environmental factors (i.e., topographic habitat, canopy height, and rock-bareness) were used as independent variables, to estimate their influences on the shifts of leaf traits. KEY RESULTS: We found that (1) plant size and the environmental factors independently drove the intraspecific leaf trait shifts of C. dunniana, of which plant size explained less variances than environmental factors. (2) With increasing plant size, C. dunniana individuals had increasingly smaller SLA but larger sized leaves. (3) The most influential environmental factor was topographic habitat; it drove the shifts of all the four traits examined. Clausena dunniana individuals on hilltops had leaf traits representing more conservative resource-use strategies (e.g., smaller SLA, higher LDMC) than individuals on slopes. On top of that, local-scale environmental factors further modified leaf trait shifts. CONCLUSIONS: Plant size and environment independently shaped the variations in intraspecific leaf traits of C. dunniana in the subtropical karst forest of Maolan. Compared with plant size, the environment played a more critical role in shaping intraspecific leaf trait variations, and potentially also the underlying individual-level plant resource-use strategies.

Have China's national forest reserves designated since 1990 conserved forests effectively?

China's forests were severely degraded by human activities during the latter half of the 20th century. Therefore, China enacted ambitious programs of natural forest protection and afforestation to protect and expand forests. Yet it is unclear how the programs, especially the designation of forest reserves, have affected forest cover and fragmentation. We evaluated the effectiveness of China's national forest reserves designated since 1990 in conserving forests, by analyzing four forest metrics (i.e., percentage forest cover, mean forest patch size, mean forest patch radius of gyration, and forest patch cohesion index) derived from a newly produced 30 m annual China land cover dataset from 1990 to 2019. We found that overall forest cover increased and fragmentation decreased from baseline years, when reserves were designated, to 2019 in both reserves and their surrounding areas, and only the increase in forest cover relative to baseline was significantly greater in reserves than in surrounding areas. The designation time of reserves under national protection had no considerable effect on changes in the four metrics, but for zonation, the core zone showed a significantly higher increase in forest patch cohesion index relative to baseline than the buffer and transition zones. Nevertheless, forest cover declined and fragmentation increased in highly forested reserves, suggesting destructive human activities and ineffective management. Thus, forest protection and regeneration programs were moderately successful. We recommend that there is significant improvement needed to ensure greater protection of existing forests and reduction of threats to promote effective management.

Influenza A and B outbreaks differed in their associations with climate conditions in Shenzhen, China.

Under the variant climate conditions in the transitional regions between tropics and subtropics, the impacts of climate factors on influenza subtypes have rarely been evaluated. With the available influenza A (Flu-A) and influenza B (Flu-B) outbreak data in Shenzhen, China, which is an excellent example of a transitional marine climate, the associations of multiple climate variables with these outbreaks were explored in this study. Daily laboratory-confirmed influenza virus and climate data were collected from 2009 to 2015. Potential impacts of daily mean/maximum/minimum temperatures (T/Tmax/Tmin), relative humidity (RH), wind velocity (V), and diurnal temperature range (DTR) were analyzed using the distributed lag nonlinear model (DLNM) and generalized additive model (GAM). Under its local climate partitions, Flu-A mainly prevailed in summer months (May to June), and a second peak appeared in early winter (December to January). Flu-B outbreaks usually occurred in transitional seasons, especially in autumn. Although low temperature caused an instant increase in both Flu-A and Flu-B risks, its effect could persist for up to 10 days for Flu-B and peak at 17 C (relative risk (RR) = 14.16, 95% CI: 7.46-26.88). For both subtypes, moderate-high temperature (28 C) had a significant but delayed effect on influenza, especially for Flu-A (RR = 26.20, 95% CI: 13.22-51.20). The Flu-A virus was sensitive to RH higher than 76%, while higher Flu-B risks were observed at both low (< 65%) and high (> 83%) humidity. Flu-A was active for a short term after exposure to large DTR (e.g., DTR = 10 C, RR = 12.45, 95% CI: 6.50-23.87), whereas Flu-B mainly circulated under stable temperatures. Although the overall wind speed in Shenzhen was low, moderate wind (2-3 m/s) was found to favor the outbreaks of both subtypes. This study revealed the thresholds of various climatic variables promoting influenza outbreaks, as well as the distinctions between the flu subtypes. These data can be helpful in predicting seasonal influenza outbreaks and minimizing the impacts, based on integrated forecast systems coupled with short-term climate models.

Functional analysis of a dihydroflavonol 4-reductase gene in Ophiorrhiza japonica (OjDFR1) reveals its role in the regulation of anthocyanin.

Dihydroflavonol 4-reductase (DFR), a key regulatory enzyme, participated in the biosynthesis of anthocyanins, proanthocyanidins and other flavonoids that essential for plant survival and human health. However, the role of this enzyme in Ophiorrhiza japonica is still unknown. Here, three putative DFR-like genes were firstly isolated from O. japonica. Phylogenetic analysis indicated that OjDFR1 was classified into DFR subgroup, while the rest two were clustered into other NADPH-dependent reductases. Then, functions of the three genes were further characterized. Expression analysis showed that OjDFR1 transcripts had strong correlations with the accumulation pattern of anthocyanin during the flower developmental, whereas other two were not, this suggested the potential roles of OjDFR1 in anthocyanin biosynthesis. Subsequently, all three clones were functionally expressed in Escherichia coli, but confirming that only OjDFR1 encode active DFR proteins that catalyzed the reduction of dihydroflavonols to leucoanthocyanidin. Consistant with the biochemical assay results, overexpressing OjDFR1 in Arabidopsis tt3-1 mutant successfully restored the deficiency of anthocyanin and proanthocyanidin, hinting its function as DFR in planta. Additionally, heterologous expression of OjDFR1 in transgenic tobacco contributed to darker flower color via up-regulating the expressions of endogenous NtANS and NtUFGT, which suggested that OjDFR1 was involved in flower color development. In summary, this study validates the functions of OjDFR1 and expands our understanding of anthocyanin biosynthesis in O. japonica.

The complete chloroplast genome sequence of Acorus gramineus (Acoraceae).

The complete chloroplast genome sequence of Acorus gramineus was assembled and characterized as a resource for future genetic studies. With a total length of 152,887 bp, the chloroplast genome comprised of a large single-copy (LSC) region of 83,005 bp, a small single-copy (SSC) region of 18,230 bp, and two inverted repeat (IR) regions of 25,826 bp. The overall GC contents of the chloroplast genome were 38.7%. A total of 115 genes were predicted, consisting of 80 protein-coding genes, 31 tRNA genes, and 4 rRNA genes. In these genes, nine genes contained one intron and two genes contained two introns. Phylogenetic analysis confirmed the position of A. gramineus within the monocots.

Bacillus amyloliquefaciens PDR1 from root of karst adaptive plant enhances Arabidopsis thaliana resistance to alkaline stress through modulation of plasma membrane H+-ATPase activity.

Exploration of native microbes is a feasible way to develop microbial agents for ecological restoration. This study was aimed to explore the impact of Bacillus amyloliquefaciens PDR1 from karst adaptive plant on the activity of root plasma membrane H+-ATPase in Arabidopsis thaliana. A. thaliana was cultured in presence or absence of B. amyloliquefaciens PDR1 and its effects on the growth were evaluated by measuring the taproot length and dry weight. The rhizosphere acidification capacity was detected by a pH indicator, a pH meter and non-invasive micro-test techniques (NMT). The nutrient uptake was performed using appropriate methods. A combination of transcriptome sequencing and real-time quantitative polymerase chain reaction (qRT-PCR) was used to measure the expression of functional genes that regulate the plasma membrane H+-ATPase activity in A. thaliana roots. Functional analysis was performed to understand how B. amyloliquefaciens regulates biological processes and metabolic pathways to strengthen A. thaliana resistance to alkaline stress. Here, we show that volatile organic compounds (VOCs) from B. amyloliquefaciens PDR1 promoted the growth and development of A. thaliana, enhanced the plasma membrane H+-ATPase activity, and affected ion absorption in Arabidopsis roots. Moreover, B. amyloliquefaciens PDR1 VOCs did not affect the expression of the gene coding for plasma membrane H+-ATPase, but affected the expression of genes regulating the activity of plasma membrane H+-ATPase. Our findings illuminate the mechanism by which B. amyloliquefaciens regulates the growth and alkaline stress resistance of A. thaliana, and lay a foundation for wide and efficient application for agricultural production and ecological protection.

Chalcone Isomerase a Key Enzyme for Anthocyanin Biosynthesis in Ophiorrhiza japonica.

Anthocyanins are distributed ubiquitously to terrestrial plants and chalcone isomerase (CHI) catalyzes the stereospecific isomerization of chalcones - a committed step in the anthocyanin biosynthesis pathway. In this study, one gene encoding CHI was isolated from Ophiorrhiza japonica and designated as OjCHI. Multiple sequence alignments and phylogenetic analysis revealed that OjCHI had the conserved CHI active site residues and was classified into type I CHI group. In order to better understand the mechanisms of anthocyanin synthesis in O. japonica, integrative analysis between metabolites and OjCHI expression was conducted. The results showed OjCHI expression matched the accumulation patterns of anthocyanins not only in different tissues but also during the flower developmental stages, suggesting the potential roles of OjCHI in the biosynthesis of anthocyanin. Then biochemical analysis indicated that recombinant OjCHI protein exhibited a typical type I CHI activity which catalyzed the production of naringenin from naringenin chalcone. Moreover, expressing OjCHI in Arabidopsis tt5 mutant restored the anthocyanins and flavonols phenotype of hypocotyl, cotyledon and seed coat, indicating its function as a chalcone isomerase in vivo. In summary, our findings reveal the in vitro as well as in vivo functions of OjCHI and provide a resource to understand the mechanism of anthocyanin biosynthesis in O. japonica.

Distinct endophytes are used by diverse plants for adaptation to karst regions.

The present study aimed at systematically investigating the endophytic communities of dominant plants in the karst ecosystem. Soil and plant materials were collected and after sequencing of the 16 s RNA, the diversity and abundance of the endophytic community structures in leaves were examined. Our results showed that abundant and diverse endogenous bacteria were associated with the leaves of common dominant plants living in the karst ecological environment. Notably, common traits and significant differences in the endophytic community structures were recorded among different plant species with different leaf grown in soils with different calcium contents. These observations implied that plants may adopt different strategies to adapt to the karst ecological environment. In addition, the endophytic bacteria associated with the leaves may be involved in different physiological strategies used by the plants to adapt to the karst ecological environment. These findings provide new avenues for developing microbial agents that could be suitable for the karst ecological environment and will provide sustainable solutions for improving the ability of plants to adapt to karst special adversities, and thus for karst geomorphological environmental protection and agricultural development.

Karst rocky desertification progress: Soil calcium as a possible driving force.

Karst rocky desertification is a severe irreversible ecosystem failure. The karst ecosystem is so fragile that it is vulnerable to environmental changes, degrading into rocky desertification. Prior studies revealed the potential connections between the soil bacterial community, the edaphic properties and the aboveground vegetation cover in the karst ecosystem. However, how these three elements affect each other and work together in propelling in the karst rocky desertification progress largely remains unexplored. To answer this question, we monitored the bacterial community variations in soils sampled from multiple sites at a successional karst rocky desertification region by sequencing the 16S rRNA V3-V4 regions. Overall, we detected 34 bacterial phyla in the karst soils, of which Proteobacteria, Actinobacteria, and Acidobacteria are the most abundant. Network analysis of the bacterial community- vegetation-edaphic property-vegetation interactions identified 6 bacterial herds that had significant correlation with soil Ca2+ and available phosphorus change during vegetation degradation. Further functional simulation of these bacterial herds unveiled the change of Ca2+ and available phosphorus might disturb the soil carbon and nitrogen metabolism, and thus weakened soil quality. In summary, we hypothesized a calcium-driven bacterial response mechanism in the karst rocky desertification progress.

Analysis of the Bacterial Communities in Two Liquors of Soy Sauce Aroma as Revealed by High-Throughput Sequencing of the 16S rRNA V4 Hypervariable Region.

Chinese liquor is one of the world's oldest distilled alcoholic beverages and an important commercial fermented product in China. The Chinese liquor fermentation process has three stages: making Daqu (the starter), stacking fermentation on the ground, and liquor fermentation in pits. We investigated the bacterial diversity of Maotai and Guotai Daqu and liquor fermentation using high-throughput sequencing of the V4 hypervariable region of the 16S rRNA gene. A total of 70,297 sequences were obtained from the Daqu samples and clustered into 17 phyla. The composition of the bacterial communities in the Daqu from these two soy sauce aroma-style Chinese liquors was the same, although some bacterial species changed in abundance. Between the Daqu and liquor fermentation samples, 12 bacterial phyla increased. The abundance of Lactobacillus and Pseudomonas increased in the liquor fermentation. This study has used high-throughput sequencing to provide new insights into the bacterial composition of the Chinese liquor Daqu and fermentation. Similarities in the distribution of bacteria in the soy sauce aroma-style Chinese liquors Daqu suggest that the abundance of bacteria might be generally concerned to other liquor.

Fluctuating selection by water level on gynoecium colour polymorphism in an aquatic plant.

BACKGROUND AND AIMS: It has been proposed that variation in pollinator preferences or a fluctuating environment can act to maintain flower colour polymorphism. These two hypotheses were tested in an aquatic monocot Butomus umbellatus (Butomaceae) with a pink or white gynoecium in the field population. METHODS: Pollinator visitation was compared in experimental arrays of equivalent flowering cymes from both colour morphs. Seed set was compared between inter- and intramorph pollination under different water levels to test the effect of fluctuating environment on seed fertility. KEY RESULTS: Overall, the major pollinator groups did not discriminate between colour morphs. Compared with the white morph, seed production in the pink morph under intermorph, intramorph and open pollination treatments was significantly higher when the water level was low but not when it was high. Precipitation in July was correlated with yearly seed production in the pink morph but not in the white morph. CONCLUSIONS: The results indicated that the two colour morphs differed in their tolerance to water level. Our study on this aquatic plant provides additional evidence to support the hypothesis that flower colour polymorphism can be preserved by environmental heterogeneity.

Discovery of gynoecium color polymorphism in an aquatic plant.

Flower color polymorphism exhibited by natural populations provides an opportunity for understanding the evolutionary mechanisms contributing to the diversity of floral morphology. However, little is known about the color polymorphism of female organs in flowering plants. Here we report gynoecium color polymorphism in Butomus umbellatus (Butomaceae), an emergent, aquatic monocot. Populations from Mishan, northeastern China comprised two morphs; gynoecia are either pink, as observed in other areas, or white. We measured floral traits and female fecundity in the two gynoecium color morphs in the field. There was no significant difference in plant height, pedicel length, and flower size including petal, sepal and gynoecium between the two morphs, but plants with pink gynoecia had wider inflorescence stalks, larger inner whorl anthers and produced more pollen and ovules than those with white gynoecia. Correspondingly, we found that seed production was significantly higher in the pink than in the white morph. This new finding suggested selection against white gynoecia in part because of low fecundity, consistent with the rarity of the white gynoecium morph in this species.