The evolution of flower-pollinator trait matching, and why do some alpine gingers appear to be mismatched?

BACKGROUND AND AIMS: Morphological matching between flower and pollinator traits has been documented in diverse plant lineages. Indeed, the matching of corolla tube length and pollinator tongue length has been cited repeatedly as a classic case of coevolution. However, there are many possible evolutionary routes to trait matching. Our aim here is both to review the evolutionary mechanisms of plant-pollinator trait matching and to investigate a specific case of trait matching/mismatching in a genus of alpine gingers. METHODS: Roscoea gingers with long corolla tubes in the western Himalayas have pollinators with correspondingly long tongues, but the match between corolla tube and pollinator tongue lengths is not seen in the eastern Himalayas. Six floral traits were measured, including corolla tube depth, an internal trait controlling pollinator access to nectar. We calculated coefficients of variation and phylogenetically controlled correlation patterns of these traits in six Roscoea species in order to gain possible insights into stabilizing selection and modularization of these traits. KEY RESULTS: The distal (nectar-containing) portion of the corolla tube exhibited lower coefficients of variations than did the basal portion. This is consistent with the hypothesis that pollinators mediate stabilizing selection on the distal, but not basal, portion of the corolla tube. This result, combined with phylogenetic data, suggests that the elevated liquid level of nectar in the distal tube evolved subsequent to dispersal into the eastern Himalayan region and loss of long-tongue pollinators. After accounting for phylogeny, corolla tube length, anther length, style length and labellum width were all intercorrelated. Corolla-tube depth was not part of this covariational module, however, suggesting separate adaptation to short-tongued pollinators. CONCLUSIONS: The reduction in functional corolla tube depth in the Roscoea appears to be related to the loss of long-tongued pollinators associated with dispersal to the eastern Himalayas and pollination by short-tongued pollinators. The apparent mismatch between floral tubes and pollinator tongues is a case of cryptic trait matching between flowers and pollinators, underscoring the importance of combining floral anatomy with pollination ecology in assessing plant-pollinator trait matching.

Chemical Synthesis of Post-Translationally Modified H2AX Reveals Redundancy in Interplay between Histone Phosphorylation, Ubiquitination, and Methylation on the Binding of 53BP1 with Nucleosomes.

The chemical synthesis of homogeneously modified histones is a powerful approach to quantitatively decipher how post-translational modifications (PTMs) modulate epigenetic events. Herein, we describe the expedient syntheses of a selection of phosphorylated and ubiquitinated H2AX proteins in a strategy integrating expressed protein hydrazinolysis and auxiliary-mediated protein ligation. These modified H2AX proteins were then used to discover that although H2AXS139 phosphorylation can enhance the binding of the DNA damage repair factor 53BP1 to either an unmodified nucleosome or that bearing a single H2AXK15ub or H4K20me2 modification, it augments 53BP1's binding only weakly to nucleosomes bearing both H2AXK15ub and H4K20me2. To better understand why such a trivalent additive effect is lacking, we solved the cryo-EM structure (3.38 Å) of the complex of 53BP1 with the H2AXK15ub/S139ph_H4K20me2 nucleosome, which showed that H2AXS139 phosphorylation distorts the interaction interface between ubiquitin and 53BP1's UDR motif. Our study revealed that there is redundancy in the interplay of multiple histone PTMs, which may be useful for controlling the dynamic distribution of effector proteins onto nucleosomes bearing different histone variants and PTMs in a time-dependent fashion, through specific cellular biochemical events.

Thioester-Assisted Sortase-A-Mediated Ligation.

Sortase A (SrtA)-mediated ligation, a popular method for protein labeling and semi-synthesis, is limited by its reversibility and dependence on the LPxTG motif, where "x" is any amino acid. Here, we report that SrtA can mediate the efficient and irreversible ligation of a protein/peptide containing a C-terminal thioester with another protein/peptide bearing an N-terminal Gly, with broad tolerance for a wide variety of LPxT-derived sequences. This strategy, the thioester-assisted SrtA-mediated ligation, enabled the expedient preparation of proteins bearing various N- or C-terminal labels, including post-translationally modified proteins such as the Ser139-phosphorylated histone H2AX and Lys9-methylated histone H3, with less dependence on the LPxTG motif. Our study validates the chemical modification of substrates as an effective means of augmenting the synthetic capability of existing enzymatic methods.

Cysteine-Aminoethylation-Assisted Chemical Ubiquitination of Recombinant Histones.

Histone ubiquitination affects the structure and function of nucleosomes through tightly regulated dynamic reversible processes. The efficient preparation of ubiquitinated histones and their analogs is important for biochemical and biophysical studies on histone ubiquitination. Here, we report the CAACU (cysteine-aminoethylation assisted chemical ubiquitination) strategy for the efficient synthesis of ubiquitinated histone analogs. The key step in the CAACU strategy is the installation of an N-alkylated 2-bromoethylamine derivative into a recombinant histone through cysteine aminoethylation, followed by native chemical ligation assisted by Seitz's auxiliary to produce mono- and diubiquitin (Ub) and small ubiquitin-like modifier (SUMO) modified histone analogs. This approach enables the rapid production of modified histones from recombinant proteins at about 1.5-6 mg/L expression. The thioether-containing isopeptide bonds in the products are chemically stable and bear only one atomic substitution in the structure, compared to their native counterparts. The ubiquitinated histone analogs prepared by CAACU can be readily reconstituted into nucleosomes and selectively recognized by relevant interacting proteins. The thioether-containing isopeptide bonds can also be recognized and hydrolyzed by deubiquitinases (DUBs). Cryo-electron microscopy (cryo-EM) of the nucleosome containing H2BKC34Ub indicated that the obtained CAACU histones were of good quality for structural studies. Collectively, this work exemplifies the utility of the CAACU strategy for the simple and efficient production of homogeneous ubiquitinated and SUMOylated histones for biochemical and biophysical studies.

Nectar supplementation changes pollinator behaviour and pollination mode in Pedicularis dichotoma: implications for evolutionary transitions.

Backgrounds and Aims: Gain or loss of floral nectar, an innovation in floral traits, has occurred in diverse lineages of flowering plants, but the causes of reverse transitions (gain of nectar) remain unclear. Phylogenetic studies show multiple gains and losses of floral nectar in the species-rich genus Pedicularis. Here we explore how experimental addition of nectar to a supposedly nectarless species, P. dichotoma, influences pollinator foraging behaviour. Methods: The liquid (nectar) at the base of the corolla tube in P. dichotoma was investigated during anthesis. Sugar components were measured by high-performance liquid chromatography. To understand evolutionary transitions of nectar, artificial nectar was added to corolla tubes and the reactions of bumble-bee pollinators to extra nectar were examined. Key Results: A quarter of unmanipulated P. dichotoma plants contained measurable nectar, with 0.01-0.49 µL per flower and sugar concentrations ranging from 4 to 39 %. The liquid surrounding the ovaries in the corolla tubes was sucrose-dominant nectar, as in two sympatric nectariferous Pedicularis species. Bumble-bees collected only pollen from control (unmanipulated) flowers of P. dichotoma, adopting a sternotribic pollination mode, but switched to foraging for nectar in manipulated (nectar-supplemented) flowers, adopting a nototribic pollination mode as in nectariferous species. This altered foraging behaviour did not place pollen on the ventral side of the bees, and sternotribic pollination also decreased. Conclusion: Our study is the first to quantify variation in nectar production in a supposedly 'nectarless' Pedicularis species. Flower manipulations by adding nectar suggested that gain (or loss) of nectar would quickly result in an adaptive behavioural shift in the pollinator, producing a new location for pollen deposition and stigma contact without a shift to other pollinators. Frequent gains of nectar in Pedicularis species would be beneficial by enhancing pollinator attraction in unpredictable pollination environments.

Safe sites of pollen placement: a conflict of interest between plants and bees?

Plant stigmas and bee pollinators are competitors for pollen. Pollen placed on a pollinator's body can be picked up by conspecific stigmas or it can be collected by the pollinator as food. Hypothetically, one solution is for pollen to be placed on 'safe sites' on the pollinator's body, sites where the pollinator cannot easily remove it, leaving the pollen for stigmas. We compared 14 sites on the bumblebee body in terms of the ability of the bee to groom off fluorescent power, a dust that may be thought of as analogous to pollen. The safest sites were along the midline of the dorsal thorax, the dorsal abdomen, and the ventral abdomen. Next, we counted how much pollen is borne on the 14 sites by bees visiting one nectariferous and three nectarless Pedicularis species. In the four species, only 7, 26, 28, and 30% of pollen found on the bees were on safe sites. Finally, we observed that the 14 sites were contacted by stigmas of the four Pedicularis species; none of the most contacted sites were safe sites. Across all four Pedicularis species, pollen is mainly positioned on sites of the bee body that were beneficial for both the plant and the bee, not on sites detrimental to either of them. Our analysis showed that the conflict of interest between flowers and bees can be solved by cooperation. Pedicularis pollen is placed where it strengthens the mutualism between plants and pollinators.

[Quality control of human chorionic gonadotrophin subunit dissociation].

Human chorionic gonadotrophin (hCG), a glycohormone widely used in treatment of infertility, is a heterodimer composed of an alpha- and a beta-subunit. The heterodimer could be dissociated during production and storage with an impact on its bioactivity. A CE-SDS method for quantitative analysis of hCG subunit dissociation was established in this study by optimization of a variety of method conditions including sample preparation buffer compositions, incubation temperature, separation voltage, and capillary temperature. This method was validated for good sensitivity, linearity, precision, and accuracy for both α- and ß-subunit. CE-SDS also showed much better precision and accuracy than SDS-PAGE. The method was successfully used in both recombinant hCG (r-hCG) produced by cell culture and hCG (u-hCG) derived from urine. The CE-SDS method was used in the study of hCG development and stability. Therefore, it is an useful tool for the quality control of hCG.

Pre- and post-pollination interaction between six co-flowering Pedicularis species via heterospecific pollen transfer.

It remains unclear how related co-flowering species with shared pollinators minimize reproductive interference, given that the degree of interspecific pollen flow and its consequences are little known in natural communities. Differences in pollen size in six Pedicularis species with different style lengths permit us to measure heterospecific pollen transfer (HPT) between species pairs in sympatry. The role of pollen-pistil interactions in mitigating the effects of HPT was examined. Field observations over 2 yr showed that bumblebee pollinators visiting one species rarely moved to another. Heterospecific pollen (HP) comprised < 10% of total stigmatic pollen loads for each species over 2 yr, and was not related to conspecific pollen deposition. Species with longer styles generally received more HP per stigma. The pollen tube study showed that pollen from short-styled species could not grow the full length of the style of long-styled species. Pollen from long-styled species could grow through the short style of P. densispica, but P. densispica rarely received HP in nature. Flower constancy is a key pre-pollination barrier to HPT between co-flowering Pedicularis species. Post-pollination pollen-pistil interactions may further mitigate the effects of HPT because HP transferred to long styles could generally be effectively filtered.

Identification of heavy metal sources in the reclaimed farmland soils of the pearl river estuary in China using a multivariate geostatistical approach.

Heavy metals in the reclaimed farmland soils of the Pearl River Estuary in China have attracted much attention because of the health risk posed to local residents. The identification of heavy metal sources in these soils is necessary to reduce their health risk. Reclaimed farmland soil samples were collected from 144 sites in the Pearl River Estuary and the contents of heavy metals (Cd, Pb, Cr, Ni, Cu, and Zn) were determined. All these heavy metals showed concentrations substantially higher than their background values, indicating possible anthropogenic pollution. The results of a multivariate geostatistical method demonstrate that grouped Cd, Cr, and Cu were mainly controlled by chemical fertilizers. Grouped Pb and Zn were the most severely impacted by atmospheric deposition from Guangzhou and Foshan, and Ni was primarily impacted by electroplating factories' wastewater discharge.

Degree of style coiling is associated with corolla-tube length in the nectarless flowers of Roscoea schneideriana.

The plant-pollinator 'arms race' model posits that a major driver of the evolution of elongated corollas in flowers is reciprocal selection for 'morphological fit' between pollinator-tongue length and access distance to nectar (usually corolla-tube length). Evidence for the pollinator-mediated selection on tube length and evolution of multiple, correlated floral traits remains inconclusive. To gain possible insights into the strength of stabilizing selection by assessing standing phenotypic variation, we measured a series of functionally important floral traits, including corolla tube length and 'effective' tube depth and degree of style coiling. We then calculated coefficients of variation (CV) for these traits in three field populations of R. schneideriana. Unlike in most long-tubed flowers, the bottom part of the corolla tube is completely occupied by the style, with no room for nectar. The length of this portion of the corolla tube was more variable (higher CV) than the upper part of the corolla tube, suggesting that functional tube depth was under stronger stabilizing selection. The degree of style coiling was negatively related to the corolla-tube length in all three populations of R. schneideriana, suggesting that there may be conflicting selection acting on style length and corolla-tube length, which are otherwise usually tightly correlated. Given the lack of nectar in the flowers of this species, the long corolla tubes and long styles may represent morphological holdovers from ancestors that were pollinated by long-tongued pollinators, as is still seen in related species in the western Himalayas.

CNTN1 Aggravates Neuroinflammation and Triggers Cognitive Deficits in Male Mice by Boosting Crosstalk between Microglia and Astrocytes.

A wealth of knowledge regarding glial cell-mediated neuroinflammation, which contributes to cognitive deficits in Alzheimer's disease (AD) has emerged in recent years. Contactin 1(CNTN1), a member of the cell adhesion molecule and immunoglobulin supergene family, is centrally involved in axonal growth regulation and is also a key player in inflammation-associated disorders. However, whether CNTN1 plays a role in inflammation-related cognitive deficits and how this process is triggered and orchestrated remain to be fully elucidated. In this study, we examined postmortem brains with AD. CNTN1 immunoreactivity was markedly increased, particularly in the CA3 subregion, as compared with non-AD brains. Furthermore, by applying an adeno-associated virus-based approach to overexpress CNTN1 directly via stereotactic injection in mice, we demonstrated that hippocampal CNTN1 overexpression triggered cognitive deficits detected by novel object-recognition, novel place-recognition and social cognition tests. The mechanisms underlying these cognitive deficits could be attributed to hippocampal microglia and astrocyte activation, which led to aberrant expression of excitatory amino acid transporters (EAAT)1/EAAT2. This resulted in long-term potentiation (LTP) impairment that could be reversed by minocyline, an antibiotic and the best-known inhibitor of microglial activation. Taken together, our results identified Cntn1 as a susceptibility factor involved in regulating cognitive deficits via functional actions in the hippocampus. This factor correlated with microglial activation and triggered astrocyte activation with abnormal EAAT1/EAAT2 expression and LTP impairment. Overall, these findings may significantly advance our understanding of the pathophysiological mechanisms underlying the risk of neuroinflammation related cognitive deficits.

The Mechanism of Chromatin Remodeler SMARCAD1/Fun30 in Response to DNA Damage.

DNA packs into highly condensed chromatin to organize the genome in eukaryotes but occludes many regulatory DNA elements. Access to DNA within nucleosomes is therefore required for a variety of biological processes in cells including transcription, replication, and DNA repair. To cope with this problem, cells employ a set of specialized ATP-dependent chromatin-remodeling protein complexes to enable dynamic access to packaged DNA. In the present review, we summarize the recent advances in the functional and mechanistic studies on a particular chromatin remodeler SMARCAD1Fun30 which has been demonstrated to play a key role in distinct cellular processes and gained much attention in recent years. Focus is given to how SMARCAD1Fun30 regulates various cellular processes through its chromatin remodeling activity, and especially the regulatory role of SMARCAD1Fun30 in gene expression control, maintenance and establishment of heterochromatin, and DNA damage repair. Moreover, we review the studies on the molecular mechanism of SMARCAD1Fun30 that promotes the DNA end-resection on double-strand break ends, including the mechanisms of recruitment, activity regulation and chromatin remodeling.

Porcine circovirus 3 in cattle in Shandong province of China: A retrospective study from 2011 to 2018.

Porcine circovirus type 3 (PCV3) is a new member of Circovirus, which could cause various symptoms in infected pigs. It has been reported in many countries and detected from various animals. This study retrospectively analyzed serum samples that were randomly collected from 1,499 clinically healthy cattle in Shandong province from 2011 to 2018. The PCV3 DNA was detected in 28.95% (434/1,499) of samples. Twenty-seven cap genes of PCV3 were sequenced and compared with seventy reference sequences. They were in several different branches, but all belonged to PCV3b. The results indicated that PCV3 was prevalent in health cattle in Shandong province of China. Though infected cattle did not show any clinical symptoms, they could be a reservoir for the virus and probably transferred them back to pigs.

Use of flue gas desulfurization gypsum for leaching Cd and Pb in reclaimed tidal flat soil.

A soil column leaching experiment was conducted to eliminate heavy metals from reclaimed tidal flat soil. Flue gas desulfurization (FGD) gypsum was used for leaching. The highest removal rates of Cd and Pb in the upper soil layers (0-30 cm) were 52.7 and 30.5 %, respectively. Most of the exchangeable and carbonate-bound Cd and Pb were removed. The optimum FGD gypsum application rate was 7.05 kg·m(-2), and the optimum leaching water amount for the application was 217.74 L·m(-2). The application of FGD gypsum (two times) and the extension of the leaching interval time to 20 days increased the heavy metal removal rate in the upper soil layers. The heavy metals desorbed from the upper soil layers were re-adsorbed and fixed in the 30-70 cm soil layers.

The stability of insulin-loaded polybutylcyanoacrylate nanoparticles in an oily medium and the hypoglycemic effect in diabetic rats.

AIM: To study the stability of insulin-loaded polybutylcyanoacrylate nanoparticles (IPN) in an oily medium (soybean oil containing 0.5% (v/v) Tween-20 and 5% (v/v) Vitamin E) along with the hypoglycemic effect following their oral administration to streptozotocin (STZ) induced diabetic rats. METHODS: The stability of IPN in the process was appraised by measurement of the amount of undegraded insulin associated to nanoparticles, the average size and the span of IPN, as well as the release of insulin from IPN. IPN in an aqueous medium (containing 0.5% (v/v) Tween-20) at pH 2.0 was also investigated as control. RESULTS: The study showed that IPN in the oily medium was more stable than that in the aqueous medium over one year of storage in the dark at (25 +/- 2) degrees C and the in vitro stability of IPN in the oily medium against degradation by proteolytic enzymes was much better than that in the aqueous medium. The apparent bioavailability of an oral administration of IPN (50 u x kg(-1)) in the oily medium versus an (sc) injection of insulin (2 u x kg(-1)) was 22.4%, much higher than that of IPN in the aqueous medium (15.5%), based on decreased areas above curve (AAC) determination for the blood glucose depression from time zero to 144 h of a single oral administration of IPN to STZ-diabetic rats. CONCLUSION: IPN in soybean oil containing Tween-20 (0.5% v/v) and Vitamin E (5% v/v) could be considered as an effective and stable delivery system for oral insulin.

Novel Nano-composites SDC-LiNaSO4 as Functional Layer for ITSOFC.

As an ionic conductive functional layer of intermediate temperature solid oxide fuel cells (ITSOFC), samarium-doped ceria (SDC)-LiNaSO4 nano-composites were synthesized by a sol-gel method and their properties were investigated. It was found that the content of LiNaSO4 strongly affected the crystal phase, defect concentration, and conductivity of the composites. When the content of LiNaSO4 was 20 wt%, the highest conductivity of the composite was found to be, respectively, 0.22, 0.26, and 0.35 S cm-1 at temperatures of 550, 600, and 700 °C, which are much higher than those of SDC. The peak power density of the single cell using this composite as an interlayer was improved to, respectively, 0.23, 0.39, and 0.88 W cm-2 at 500, 600, and 700 °C comparing with that of the SDC-based cell. Further, the SDC-LiNaSO4(20 wt%)-based cell also displayed better thermal stability according to the performance measurements at 560 °C for 50 h. These results reveal that SDC-LiNaSO4 composite may be a potential good candidate as interlayer for ITSOFC due to its high ionic conductivity and thermal stability.

Quality control of human chorionic gonadotrophin subunit dissociation / 药学学报

Human chorionic gonadotrophin (hCG), a glycohormone widely used in treatment of infertility, is a heterodimer composed of an alpha-and a beta-subunit. The heterodimer could be dissociated during production and storage with an impact on its bioactivity. A CE-SDS method for quantitative analysis of hCG subunit dissociation was established in this study by optimization of a variety of method conditions including sample preparation buffer compositions, incubation temperature, separation voltage, and capillary temperature. This method was validated for good sensitivity, linearity, precision, and accuracy for both α-and β-subunit. CE-SDS also showed much better precision and accuracy than SDS-PAGE. The method was successfully used in both recombinant hCG (r-hCG) produced by cell culture and hCG (u-hCG) derived from urine. The CE-SDS method was used in the study of hCG development and stability. Therefore, it is an useful tool for the quality control of hCG.