SUMMARYMutant analyses in different eudicotyledonous flowering plants demonstrated that SEPALLATA-like MADSbox genes are required for the specification of sepals, petals, stamens and carpels, and for floral determinacy, thus defining class E floral organ identity genes. SEP-like genes encode MADS-domain transcription factors and constitute an angiosperm-specific gene clade whose members show remarkably different degrees of redundancy and sub-functionalization within eudicots. To better understand the evolutionary dynamics of SEP-like genes throughout the angiosperms we have knocked down SEP-like genes of rice (Oryza sativa), a distant relative of eudicots within the flowering plants. Plants affected in both OsMADS7 and OsMADS8 show severe phenotypes including late flowering, homeotic changes of lodicules, stamens and carpels into palea/ lemma-like organs, and a loss of floral determinacy. Simultaneous knockdown of the four rice SEP-like genes OsMADS1, OsMADS5, OsMADS7 and OsMADS8, leads to homeotic transformation of all floral organs except the lemma into leaf-like organs. This mimics the phenotype observed with the sep1 sep2 sep3 sep4 quadruple mutant of Arabidopsis. Detailed analyses of the spatial and temporal mRNA expression and protein interaction patterns corresponding to the different rice SEP-like genes show strong similarities, but also gene-specific differences. These findings reveal conservation of SEP-like genes in specifying floral determinacy and organ identities since the separation of eudicots and monocots about 150 million years ago. However, they indicate also monocot-specific neo-and sub-functionalization events and hence underscore the evolutionary dynamics of SEP-like genes. Moreover, our findings corroborate the view that the lodicules of grasses are homologous to eudicot petals.

Floral MADS-box genes encode transcription factors that play critical roles in the development and evolution of the flower. Proteins of floral MADS-box genes regulate the expression of their downstream genes by forming various homodimers/heterodimers and quaternary complexes. Interactions among proteins of floral MADS-box genes have been documented in several model species, yet the information accumulated so far is still not sufficient to draw a general picture of the evolution of the interactions. We have characterized 28 putative floral MADS-box genes from three representative basal eudicots (i.e., Euptelea pleiospermum, Akebia trifoliata, and Pachysandra terminalis) and investigated the protein-protein interactions (PPIs) among the proteins encoded by these genes using yeast two-hybrid assays. We found that, although the PPIs in basal eudicots are largely consistent with those in core eudicots and monocots, there are lineage-specific features that have not been observed elsewhere. We also reconstructed the evolutionary histories of the PPIs among members of seven MADS-box gene lineages (i.e., AP1, AP3, PI, AG, STK, AGL2, and AGL9) in angiosperms. We revealed that the PPIs were extremely conserved in nine (or 32.1%) of the 28 possible combinations, whereas considerable variations existed in seven (25.0%) of them; in the remaining 12 (or 42.9%) combinations, however, no interaction was observed. Notably, most of the PPIs required for the formation of quaternary complexes, as suggested by the "quartet model," were highly conserved. This suggested that the evolutionarily conservative PPIs may have played critical roles in the establishment of the basic structure (or architecture) of the flower and experienced coevolution to maintain their functions. The evolutionarily variable PPIs, however, seem to have played subsidiary roles in flower development and have contributed to the variation in floral traits.

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There is evidence that gene duplication and diversification within the MADS-box gene family had significant impact on floral architecture. In this study, we report the isolation of four class B homologous genes from Akebia trifoliata, termed AktAP3_1, AktAP3_2, AktAP3_3, and AktPI. Phylogenetic analysis indicates that the three AktAP3 paralogs were produced by two gene duplication events and AktAP3_2 and AktAP3_3 are recent paralogs, which are yielded by the duplication before the origin of the genus Akebia. In situ hybridization demonstrates that these genes are mainly expressed in the stamens and carpels of A. trifoliata, but in differential patterns, similar to those in other basal eudicot and basal angiosperm species. AktAP3_3 and AktPI are expressed in the developing petaloid perianth, suggesting that the petaloidy of the perianth is caused by the expression of class B genes. Reverse transcriptase polymerase chain reaction analyses indicate that these genes are expressed in both male and female flowers, but at different levels. We explore the interaction behavior of the class B proteins in the basal eudicots using yeast two-hybrid system for the first time. The AktAP3_1/2/3 proteins and the AktPI protein can form obligate heterodimers, but at different strength. From the mRNA expression and protein interaction patterns of the duplicated copies of the AktAP3 genes, we conclude that subfunctionalization very likely contributes to the maintenance of multiple AP3-like gene copies in A. trifoliata.

Summary• In core eudicots, euAP3-type MADS-box genes encode a PISTILLATA (PI)-derived motif, as well as a C-terminal euAP3 motif that originated from a paleoAP3 motif of an ancestral APETALA3 (AP3)-like protein through a translational frameshift mutation. To determine the functional and evolutionary relevance of these motifs, a series of point mutation and domain-swap constructs were generated, involving CsAP3, a paleoAP3-type gene from the basal angiosperm Chloranthus spicatus encoding a truncated paleo-AP3 motif, and AtAP3, a euAP3-type gene from the core eudicot Arabidopsis thaliana.• The chimeric constructs were expressed in A. thaliana under the control of the AP3 promoter or the CaMV 35S promoter in an ap3 mutant or wild-type background, respectively.• Significant recovery of AP3 function was obtained in both complementation and ectopic expression experiments whenever the region upstream of the C-terminal motifs (MIK region) from A. thaliana was taken, even when the PI-derived motif and the truncated paleoAP3 motif of CsAP3 substituted for the corresponding sequences from AtAP3. However, no or very weak complementation or gain-of-function was seen when the MIK region was from CsAP3.• Our data suggest that changes in the MIK region rather than mutations in the C-terminal domain were of crucial importance for the evolution of the functional specificity of euAP3-type proteins in stamen and petal development.

13C NMR signals of PCF, PBF and PBCFs co-polyesters.

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Fuel cells and water splitting are promising sustainable energy storage and conversion systems that can facilitate the usage of renewable resources and reduce the reliance on fossil fuels. These applications require catalysts to perform the required oxygen reduction reaction (ORR), oxygen evolution reaction (OER), and hydrogen evolution reaction (HER). It would be ideal to use a non-noble metal-based multifunctional catalyst for these reactions. Herein, a new type of porous carbon doped with Cu, N, S was prepared as a trifunctional catalyst for the ORR, OER, and HER from S-rich polyphenylene sulfide (PPS). By oxidatively treating the PPS, we critically prevented high-temperature melting of the precursor. Further, high-temperature pyrolysis using ammonia (NH 3 ) desulfurized and introduced N into the carbon matrix, increasing structural defects and the surface area. By introducing copper during the pyrolysis, the tridoped (Cu, N, S) catalyst was successfully synthesized. The extremely large number of active sites and the local chemical environment enable excellent electrocatalytic performance and stability across ORR, OER, and HER at levels superior or comparable to noble metals. This catalyst was also used as the sole catalyst in Zn−air batteries and overall water splitting to demonstrate excellent performance. This synthesis method can pave the way toward new catalyst design and discovery, enabling more versatile and robust energy applications.