A new brood-pollination mutualism between Stellera chamaejasme and flower thrips Frankliniella intonsa.

BACKGROUND: Brood pollination mutualism is a special type of plant-pollinator interaction in which adult insects pollinate plants, and the plants provide breeding sites for the insects as a reward. To manifest such a mutualism between Stellera chamaejasme and flower thrips of Frankliniella intonsa, the study tested the mutualistic association of the thrips life cycle with the plant flowering phenology and determined the pollination effectiveness of adult thrips and their relative contribution to the host's fitness by experimental pollinator manipulation. RESULTS: The adult thrips of F. intonsa, along with some long-tongue Lepidoptera, could serve as efficient pollinators of the host S. chamaejasme. The thrips preferentially foraged half-flowering inflorescences of the plants and oviposited in floral tubes. The floral longevity was 11.8 ± 0.55 (mean ± se) days, which might precisely accommodate the thrips life cycle from spawning to prepupation. The exclusion of adult thrips from foraging flowers led to a significant decrease in the fitness (i.e., seed set) of host plants, with a corresponding reduction in thrips fecundity (i.e., larva no.) in the flowers. CONCLUSIONS: The thrips of F. intonsa and the host S. chamaejasme mutualistically interact to contribute to each other's fitness such that the thrips pollinate host plants and, as a reward, the plants provide the insects with brooding sites and food, indicating the coevolution of the thrips life cycle and the reproductive traits (e.g., floral longevity and morphology) of S. chamaejasme.

The expansion process of a Stellera chamaejasme population in a degraded alpine meadow of Northwest China.

The expansion of poisonous plants can change vegetation community structures and affect grassland ecosystem service values. Stellera chamaejasme is one of the most important poisonous plants and has rapidly expanded in the arid areas of Northwest China in recent decades. The objective of this study was to elucidate the expansion process and model of an S. chamaejasme population. Therefore, we classified the S. chamaejasme population into five classes based on coverage: 31-40%, 41-50%, 51-60%, 61-70% and 71-80%. We investigated the spatial distribution patterns and the size compositions of S. chamaejasme under different coverages. The results show that the spatial distribution pattern of S. chamaejasme under low coverage (31-40%) at all study scales (0-100 cm) was random; the spatial distribution pattern translated to a clumped distribution from a random distribution at some scales, and the clumped distributions gradually became obvious, with coverage increasing from 41-50% to 61-70%; the spatial distribution tended to be random at all study scales when coverage was increased further (71-80%). However, the spatial distribution patterns were closely related to the size composition of the S. chamaejasme population. In particular, the quantity of older individuals had a significant impact on the variation of the spatial distribution patterns of S. chamaejasme. The spatial distribution pattern varied from a random distribution to a clumped distribution and then returned to a random distribution with increasing coverage (from 31-40% to 71-80%), and this may indicate that the S. chamaejasme patches experienced patch formation and extension and merged with each other.

H2O2 and NADPH oxidases involve in regulation of 2-(2-phenylethyl)chromones accumulation during salt stress in Aquilaria sinensis calli.

2-(2-Phenylethyl)chromones are the main compounds responsible for the quality of agarwood, which is widely used in traditional medicines, incenses and perfumes. H2O2 and NADPH oxidases (also known as respiratory burst oxidase homologs, Rbohs) mediate diverse physiological and biochemical processes in environmental stress responses. However, little is known about the function of H2O2 and NADPH oxidases in 2-(2-phenylethyl)chromones accumulation. In this study, we found that salt stress induced a transient increase in content of H2O2 and 2-(2-phenylethyl)chromones accumulation in Aquilaria sinensis calli. Exogenous H2O2 remarkably decreased the production of 2-(2-phenylethyl)chromones, while dimethylthiourea (DMTU), a scavenger of H2O2, significantly increased 2-(2-phenylethyl)chromones accumulation in salt treated calli. Three new H2O2-generating genes, named AsRbohA-C, were isolated and characterized from A. sinensis. Salt stress also induced a transient increase in AsRbohA-C expression and NADPH oxidase activity. Furthermore, exogenous H2O2 increased AsRbohA-C expression and NADPH oxidase activity, while DMTU inhibited AsRbohA-C expression and NADPH oxidase activity under salt stress. Moreover, diphenylene iodonium (DPI), the inhibitor of NADPH oxidases, reduced AsRbohA-C expression and NADPH oxidase activity, but significantly induced 2-(2-phenylethyl)chromones accumulation during salt stress. These results clearly demonstrated the central role of H2O2 and NADPH oxidases in regulation of salt-induced 2-(2-phenylethyl)chromones accumulation in A. sinensis calli.

Transcriptome Sequencing of Chemically Induced Aquilaria sinensis to Identify Genes Related to Agarwood Formation.

BACKGROUND: Agarwood is a traditional Chinese medicine used as a clinical sedative, carminative, and antiemetic drug. Agarwood is formed in Aquilaria sinensis when A. sinensis trees are threatened by external physical, chemical injury or endophytic fungal irritation. However, the mechanism of agarwood formation via chemical induction remains unclear. In this study, we characterized the transcriptome of different parts of a chemically induced A. sinensis trunk sample with agarwood. The Illumina sequencing platform was used to identify the genes involved in agarwood formation. METHODOLOGY/PRINCIPAL FINDINGS: A five-year-old Aquilaria sinensis treated by formic acid was selected. The white wood part (B1 sample), the transition part between agarwood and white wood (W2 sample), the agarwood part (J3 sample), and the rotten wood part (F5 sample) were collected for transcriptome sequencing. Accordingly, 54,685,634 clean reads, which were assembled into 83,467 unigenes, were obtained with a Q20 value of 97.5%. A total of 50,565 unigenes were annotated using the Nr, Nt, SWISS-PROT, KEGG, COG, and GO databases. In particular, 171,331,352 unigenes were annotated by various pathways, including the sesquiterpenoid (ko00909) and plant-pathogen interaction (ko03040) pathways. These pathways were related to sesquiterpenoid biosynthesis and defensive responses to chemical stimulation. CONCLUSIONS/SIGNIFICANCE: The transcriptome data of the different parts of the chemically induced A. sinensis trunk provide a rich source of materials for discovering and identifying the genes involved in sesquiterpenoid production and in defensive responses to chemical stimulation. This study is the first to use de novo sequencing and transcriptome assembly for different parts of chemically induced A. sinensis. Results demonstrate that the sesquiterpenoid biosynthesis pathway and WRKY transcription factor play important roles in agarwood formation via chemical induction. The comparative analysis of the transcriptome data of agarwood and A. sinensis lays the foundation for elucidating the mechanism of agarwood formation via chemical induction, and thus, enables future improvements in agarwood quality while protecting endangered wild A. sinensis.

Incorporating detectability of threatened species into environmental impact assessment.

Environmental impact assessment (EIA) is a key mechanism for protecting threatened plant and animal species. Many species are not perfectly detectable and, even when present, may remain undetected during EIA surveys, increasing the risk of site-level loss or extinction of species. Numerous methods now exist for estimating detectability of plants and animals. Despite this, regulations concerning survey protocol and effort during EIAs fail to adequately address issues of detectability. Probability of detection is intrinsically linked to survey effort; thus, minimum survey effort requirements are a useful way to address the risks of false absences. We utilized 2 methods for determining appropriate survey effort requirements during EIA surveys. One method determined the survey effort required to achieve a probability of detection of 0.95 when the species is present. The second method estimated the survey effort required to either detect the species or reduce the probability of presence to 0.05. We applied these methods to Pimelea spinscens subsp. spinescens, a critically endangered grassland plant species in Melbourne, Australia. We detected P. spinescens in only half of the surveys undertaken at sites where it was known to exist. Estimates of the survey effort required to detect the species or demonstrate its absence with any confidence were much higher than the effort traditionally invested in EIA surveys for this species. We argue that minimum survey requirements be established for all species listed under threatened species legislation and hope that our findings will provide an impetus for collecting, compiling, and synthesizing quantitative detectability estimates for a broad range of plant and animal species.

The sesquiterpene biosynthesis and vessel-occlusion formation in stems of Aquilaria sinensis (Lour.) Gilg trees induced by wounding treatments without variation of microbial communities.

As widely recognized, agarwood formation in Aquilaria trees is induced by external wounding. Because agarwood usually harbors specific microbes, the function of microbes in agarwood formation has been debated for almost a century. In this study, two wounding methods, the burning-chisel-drilling method (BCD) and the whole-tree agarwood-inducing method (Agar-Wit), were used under the non-contamination of environmental microorganisms. After pyrosequencing the small rRNA subunits of the wounds induced by the BCD and Agar-Wit, no substantial variation was observed either in fungal and bacterial enrichment and diversity or in the relative abundances of taxa. By contrast, significant variations in fungal and bacterial communities were detected following the partial tree pruning (PTP)-wounding. The wound-induced sesquiterpene biosynthesis and vessel-occlusion formation, however, were found to be similar in all types of wounded trunks. We thus infer that wounding in the absence of variations in microbial communities may induce agarwood formation. This result does not support the long-standing notion that agarwood formation depends on microbes.

Profiling of microRNAs under wound treatment in Aquilaria sinensis to identify possible microRNAs involved in agarwood formation.

Agarwood, a kind of highly valued non-timber product across Asia, is formed only when its resource trees--the endangered genus Aquilaria are wounded or infected by some microbes. To promote the efficiency of agarwood production and protect the wild resource of Aquilaria species, we urgently need to reveal the regulation mechanism of agarwood formation. MicroRNAs (miRNAs) are a group of gene expression regulators with overwhelming effects on a large spectrum of biological processes. However, their roles in agarwood formation remain unknown. This work aimed at identifying possible miRNAs involved in the wound induced agarwood formation. In this study, the high-throughput sequencing was adopted to identify miRNAs and monitor their expression under wound treatment in the stems of A. sinensis. The miR171, miR390, miR394, miR2111, and miR3954 families remained at the reduced level two days after the treatment. 131 homologous miRNAs in the 0.5 h library showed over three-fold variation of read number compared with the control library, of which 12 exhibiting strong expression alterations were further confirmed by real-time quantitative PCR. Target prediction and annotation of the miRNAs demonstrated that the binding, metabolic process, catalytic activity, and cellular process are the most common functions of the predicted targets of these newly identified miRNAs in A.sinensis. The cleaveage sites of three newly predicted targets were verified by 5'RACE.

A piece of the puzzle: a method for comparing pollination quality and quantity across multiple species and reproductive events.

• Understanding how pollination affects plant reproductive success and how changes in pollination service affect plant populations, communities and ecosystems is of increasing concern. Yet supplemental hand-pollination traditionally used to assess pollen limitation is prohibitive for large-scale comparative work. Moreover, it does not differentiate between quality and quantity aspects of pollen limitation, and it may suffer from confounds of post-pollination processes such as resource availability to fill seeds. • Here, we highlight pollen tubes as the functional link between pollen arrival and seed production and suggest that consideration of pollen tubes leads to a better depiction of limitation at the pre-zygotic (pollination) phase of sexual reproduction. • We assessed the rigor of piecewise regression to analyze the relationship between the numbers of pollen grains and pollen tubes observed in nonmanipulated wilted flowers. We illustrate how parameters obtained from this analysis provide quantitative insight into the relative relevance of the quantity and quality of pollen receipt in limiting natural pollination success, and can facilitate comparisons among data sets. • This nonmanipulative method opens up new opportunities for rigorous assessment of the relative importance of the quantity and quality of pollination in limiting plant reproduction, especially from a community-wide perspective.