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This study addresses the morphological similarities and taxonomic complexity of the genus Tarenaya Raf. of the family Cleomaceae Bercht. & J. Presl as well as the medicinal use of their species. The research compares potential anatomical diagnostic characters of the vegetative organs of species Tarenaya aculeata, Tarenaya diffusa, and Tarenaya spinosa to determine authenticity parameters. The study also carried out histochemical and phytochemical analyses of leaf blades to explore the medicinal use of these species. Semipermanent slides containing cross sections of the stems, petioles, and leaf blades, as well as paradermal sections of leaf blades, were prepared. The analyses were carried out in light and polarized optical microscopy. The histochemical analysis included different reagents depending on the target metabolite, which were analyzed by optical and fluorescence microscopy. Phytochemical tests of the methanolic extracts of the leaves were performed using thin layer chromatography. Anatomical characterization showed the characters of general occurrence in the family Cleomaceae and those that allow the differentiation of T. aculeata, T. diffusa, and T. spinosa. Histochemistry revealed the synthesis and/or storage sites of the metabolites and phytochemical tests; it was possible to observe the presence of different metabolites. The results bring comparative data on the anatomical and histochemical characterization of the species, thus increasing the taxonomic knowledge of the genus Tarenaya. Research Highlights: The comparative anatomy of three Tarenaya from Brazil was studied.Anatomical differences in different vegetative organs differ species.Differences in histochemistry and phytochemistry. [ABSTRACT FROM AUTHOR]

Abstract The protectorate of Saint Katherine considers one of the biggest protected areas in Egypt characterized by a unique location and environment. It supports a high number of rare and endangered species. Cleome droserifolia (Rorida droserifolia) is an endangered plant found in Saint Katherine, Sinai, Egypt. It has been known for its medicinal uses so it becomes rare due to over-grazing and over-collection for use in traditional medicine, research and trade home use. DNA barcoding analysis and in vitro culture tools were conducted for saving the plant. This research supports the goal number 15 of the United Nations Sustainable Development (SDGs); aimed at preserving, restore and reinstate sustainable usage of terrestrial ecosystems and halting biodiversity deterioration. Two pairs primers of three DNA markers: [ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit (rbcL), maturase K (matK) and trnH-psbA] were used for PCR amplification and sequencing. BLAST results and phylogenetic analysis proved a correct authentication of Cleome droserifolia on both; species and genus levels and confirms the involvement of Cleome droserifolia in Cleome genus. For in vitro propagation, cotyledonary node explants of germinated seedlings were cultured on Murashige and Skoog (MS) medium fortified with 0.27 µM α-naphthaleneacetic acid (NAA), and 2.32 µM of kinetin (Kin). The highest mean number of axillary shoots was 8.16 shoots/explant. 85% of multiple shoots were successfully rooted on ¼ MS medium fortified with 7.42 µM indole-3-butyric acid (IBA). The successfully rooted plants were transferred to a mixture of sand, soil, and peat moss (1: 1: 1) and efficiently acclimatized in the greenhouse.

The protectorate of Saint Katherine considers one of the biggest protected areas in Egypt characterized by a unique location and environment. It supports a high number of rare and endangered species. Cleome droserifolia (Rorida droserifolia) is an endangered plant found in Saint Katherine, Sinai, Egypt. It has been known for its medicinal uses so it becomes rare due to over-grazing and over-collection for use in traditional medicine, research and trade home use. DNA barcoding analysis and in vitro culture tools were conducted for saving the plant. This research supports the goal number 15 of the United Nations Sustainable Development (SDGs); aimed at preserving, restore and reinstate sustainable usage of terrestrial ecosystems and halting biodiversity deterioration. Two pairs primers of three DNA markers: [ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit (rbcL), maturase K (matK) and trnH-psbA] were used for PCR amplification and sequencing. BLAST results and phylogenetic analysis proved a correct authentication of Cleome droserifolia on both; species and genus levels and confirms the involvement of Cleome droserifolia in Cleome genus. For in vitro propagation, cotyledonary node explants of germinated seedlings were cultured on Murashige and Skoog (MS) medium fortified with 0.27 µM α-naphthaleneacetic acid (NAA), and 2.32 µM of kinetin (Kin). The highest mean number of axillary shoots was 8.16 shoots/explant. 85% of multiple shoots were successfully rooted on ¼ MS medium fortified with 7.42 µM indole-3-butyric acid (IBA). The successfully rooted plants were transferred to a mixture of sand, soil, and peat moss (1: 1: 1) and efficiently acclimatized in the greenhouse.Article highlights: Two ex situ conservation approaches for the medicinal endangered Cleome droserifolia plant were successfully conducted for saving the plant from extinction. DNA barcoding was successfully used to identify the plant at the genus and species levels. Cotyledonary node explants of germinated seedlings were used for in vitro propagation of the plant with eighty one induction percentage. This research contributes to one of the Sustainable Development Goals (SDGs) (goal number 15) by promoting the conservation, restoration and sustainable use of terrestrial ecosystems. [ABSTRACT FROM AUTHOR]

Introduction: C4 photosynthesis is an adaptation that has independently evolved at least 66 times in angiosperms. C4 plants, unlike their C3 ancestral, have a carbon concentrating mechanism which suppresses photorespiration, often resulting in faster photosynthetic rates, higher yields, and enhanced water use efficiency. Moreover, the presence of C4 photosynthesis greatly alters the relation between CO2 assimilation and stomatal conductance. Previous papers have suggested that the adjustment involves a decrease in stomatal density. Here, we tested if C4 species also have differing stomatal responses to environmental cues, to accommodate the modified CO2 assimilation patterns compared to C3 species. Methods: To test this hypothesis, stomatal responses to blue and red-light were analysed in three phylogenetically linked pairs of C3 and C4 species from the Cleomaceae (Gynandropsis and Tarenaya), Flaveria, and Alloteropsis, that use either C3 or C4 photosynthesis. Results: The results showed strongly decreased stomatal sensitivity to blue light in C4 dicots, compared to their C3 counterparts, which exhibited significant blue light responses. In contrast, in C3 and C4 subspecies of the monocot A. semialata, the blue light response was observed regardless of photosynthetic type. Further, the quantitative red-light response varied across species, but the presence or absence of a significant stomatal red-light response was not directly associated with differences in photosynthetic pathway. Interestingly, stomatal density and morphology patterns observed across the three comparisons were also not consistent with patterns commonly asserted for C3 and C4 species. Discussion: The strongly diminished blue-light sensitivity of stomatal responses in C4 species across two of the comparisons suggests a common C4 feature that may have functional implications. Altogether, the strong prevalence of speciesspecific effects clearly emphasizes the importance of phylogenetic controls in comparisons between C3 and C4 photosynthetic pathways. [ABSTRACT FROM AUTHOR]

IntroductionC4 photosynthesis is an adaptation that has independently evolved at least 66 times in angiosperms. C4 plants, unlike their C3 ancestral, have a carbon concentrating mechanism which suppresses photorespiration, often resulting in faster photosynthetic rates, higher yields, and enhanced water use efficiency. Moreover, the presence of C4 photosynthesis greatly alters the relation between CO2 assimilation and stomatal conductance. Previous papers have suggested that the adjustment involves a decrease in stomatal density. Here, we tested if C4 species also have differing stomatal responses to environmental cues, to accommodate the modified CO2 assimilation patterns compared to C3 species.MethodsTo test this hypothesis, stomatal responses to blue and red-light were analysed in three phylogenetically linked pairs of C3 and C4 species from the Cleomaceae (Gynandropsis and Tarenaya), Flaveria, and Alloteropsis, that use either C3 or C4 photosynthesis.ResultsThe results showed strongly decreased stomatal sensitivity to blue light in C4 dicots, compared to their C3 counterparts, which exhibited significant blue light responses. In contrast, in C3 and C4 subspecies of the monocot A. semialata, the blue light response was observed regardless of photosynthetic type. Further, the quantitative red-light response varied across species, but the presence or absence of a significant stomatal red-light response was not directly associated with differences in photosynthetic pathway. Interestingly, stomatal density and morphology patterns observed across the three comparisons were also not consistent with patterns commonly asserted for C3 and C4 species.DiscussionThe strongly diminished blue-light sensitivity of stomatal responses in C4 species across two of the comparisons suggests a common C4 feature that may have functional implications. Altogether, the strong prevalence of species-specific effects clearly emphasizes the importance of phylogenetic controls in comparisons between C3 and C4 photosynthetic pathways.

Premise: Flowering plants have evolved a vast array of floral features involved in plant–pollinator interactions. A feature that seemingly increases the chance of pollen transfer is the androgynophore, a stalk‐like structure that raises the reproductive organs of the flower. However, little is known about the developmental and genetic basis of this structure despite its presence in multiple, distantly related taxa. Here, we address this gap by investigating Gynandropsis gynandra (Cleomaceae), a species with a prominent androgynophore. Methods: We combined morphological and anatomical analyses with a comparative transcriptomic study to provide a detailed description of the androgynophore throughout development, examine global gene expression patterns, and identify candidate genes putatively involved in androgynophore elongation. Results: The radially symmetric androgynophore of G. gynandra rapidly lengthens primarily via cell elongation. Despite its structural uniformity, androgynophore development is characterized by complex gene expression patterns including differential expression of floral organ identity genes and genes associated with organ development and growth in Arabidopsis thaliana. Conclusions: Our morphological characterizations and high‐quality transcriptome for G. gynandra suggest that the androgynophore is a novel structure formed via elaboration of both the receptacle and base of reproductive organs because it is structurally like an elongated internode but expresses the genetic repertoire typically associated with the reproductive organs. The drastic increase in cell length and uniform structure elevates the androgynophore as a potentially powerful model for cell elongation. [ABSTRACT FROM AUTHOR]

Floral nectaries have evolved multiple times and rapidly diversified with the adaptive radiation of animal pollinators. As such, floral nectaries exhibit extraordinary variation in location, size, shape, and secretory mechanism. Despite the intricate ties to pollinator interactions, floral nectaries are often overlooked in morphological and developmental studies. As Cleomaceae exhibits substantial floral diversity, our objective was to describe and compare floral nectaries between and within genera. Floral nectary morphology was assessed through scanning electron microscopy and histology across three developmental stages of nine Cleomaceae species including representatives for seven genera. A modified fast green and safranin O staining protocol was used to yield vibrant sections without highly hazardous chemicals. Cleomaceae floral nectaries are most commonly receptacular, located between the perianth and stamens. The floral nectaries are supplied by vasculature, often contain nectary parenchyma, and have nectarostomata. Despite the shared location, components, and secretory mechanism, the floral nectaries display dramatic diversity in size and shape, ranging from adaxial protrusions or concavities to annular disks. Our data reveal substantive lability in form with both adaxial and annular floral nectaries interspersed across Cleomaceae. Floral nectaries contribute to the vast morphological diversity of Cleomaceae flowers and so are valuable for taxonomic descriptions. Though Cleomaceae floral nectaries are often derived from the receptacle and receptacular nectaries are common across flowering plants, the role of the receptacle in floral evolution and diversification is overlooked and warrants further exploration. [ABSTRACT FROM AUTHOR]

Nectaries are a promising frontier for plant evo-devo research, and are particularly fascinating given their diversity in form, position, and secretion methods across angiosperms. Emerging model systems permit investigations of the molecular basis for nectary development and nectar secretion across a range of taxa, which addresses fundamental questions about underlying parallelisms and convergence. Herein, we explore nectary development and nectar secretion in the emerging model taxa, Cleome violacea (Cleomaceae), which exhibits a prominent adaxial nectary. First, we characterized nectary anatomy and quantified nectar secretion to establish a foundation for quantitative and functional gene experiments. Next, we leveraged RNA-seq to establish gene expression profiles of nectaries across three key stages of development: pre-anthesis, anthesis, and post-fertilization. We then performed functional studies on five genes that were putatively involved in nectary and nectar formation: CvCRABSCLAW (CvCRC), CvAGAMOUS (CvAG), CvSHATTERPROOF (CvSHP), CvSWEET9, and a highly expressed but uncharacterized transcript. These experiments revealed a high degree of functional convergence to homologues from other core Eudicots, especially Arabidopsis. CvCRC, redundantly with CvAG and CvSHP, are required for nectary initiation. Concordantly, CvSWEET9 is essential for nectar formation and secretion, which indicates that the process is eccrine based in C. violacea. While demonstration of conservation is informative to our understanding of nectary evolution, questions remain. For example, it is unknown which genes are downstream of the developmental initiators CvCRC, CvAG, and CvSHP, or what role the TCP gene family plays in nectary initiation in this family. Further to this, we have initiated a characterization of associations between nectaries, yeast, and bacteria, but more research is required beyond establishing their presence. Cleome violacea is an excellent model for continued research into nectary development because of its conspicuous nectaries, short generation time, and close taxonomic distance to Arabidopsis. [ABSTRACT FROM AUTHOR]

Nectaries are a promising frontier for plant evo-devo research, and are particularly fascinating given their diversity in form, position, and secretion methods across angiosperms. Emerging model systems permit investigations of the molecular basis for nectary development and nectar secretion across a range of taxa, which addresses fundamental questions about underlying parallelisms and convergence. Herein, we explore nectary development and nectar secretion in the emerging model taxa, Cleome violacea (Cleomaceae), which exhibits a prominent adaxial nectary. First, we characterized nectary anatomy and quantified nectar secretion to establish a foundation for quantitative and functional gene experiments. Next, we leveraged RNA-seq to establish gene expression profiles of nectaries across three key stages of development: pre-anthesis, anthesis, and post-fertilization. We then performed functional studies on five genes that were putatively involved in nectary and nectar formation: CvCRABSCLAW (CvCRC), CvAGAMOUS (CvAG), CvSHATTERPROOF (CvSHP), CvSWEET9, and a highly expressed but uncharacterized transcript. These experiments revealed a high degree of functional convergence to homologues from other core Eudicots, especially Arabidopsis. CvCRC, redundantly with CvAG and CvSHP, are required for nectary initiation. Concordantly, CvSWEET9 is essential for nectar formation and secretion, which indicates that the process is eccrine based in C. violacea. While demonstration of conservation is informative to our understanding of nectary evolution, questions remain. For example, it is unknown which genes are downstream of the developmental initiators CvCRC, CvAG, and CvSHP, or what role the TCP gene family plays in nectary initiation in this family. Further to this, we have initiated a characterization of associations between nectaries, yeast, and bacteria, but more research is required beyond establishing their presence. Cleome violacea is an excellent model for continued research into nectary development because of its conspicuous nectaries, short generation time, and close taxonomic distance to Arabidopsis.

Premise: Cleomaceae is an important model clade for studies of evolutionary processes including genome evolution, floral form diversification, and photosynthetic pathway evolution. Diversification and divergence patterns in Cleomaceae remain tangled as research has been restricted by its worldwide distribution, limited genetic sampling and species coverage, and a lack of definitive fossil calibration points., Methods: We used target sequence capture and the Angiosperms353 probe set to perform a phylogenetic study of Cleomaceae. We estimated divergence times and biogeographic analyses to explore the origin and diversification of the family. Seed morphology across extant taxa was documented with multifocal image-stacking techniques and morphological characters were extracted, analyzed, and compared to fossil records., Results: We recovered a well-supported and resolved phylogenetic tree of Cleomaceae generic relationships that includes 236 (~86%) species. We identified 11 principal clades and confidently placed Cleomella as sister to the rest of the family. Our analyses suggested that Cleomaceae and Brassicaceae diverged ~56 mya, and Cleomaceae began to diversify ~53 mya in the Palearctic and Africa. Multiple transatlantic disjunct distributions were identified. Seeds were imaged from 218 (~80%) species in the family and compared to all known fossil species., Conclusions: Our results represent the most comprehensive phylogenetic study of Cleomaceae to date. We identified transatlantic disjunctions and proposed explanations for these patterns, most likely either long-distance dispersals or contractions in latitudinal distributions caused by climate change over geological timescales. We found that seed morphology varied considerably but mostly mirrored generic relationships., (© 2024 The Author(s). American Journal of Botany published by Wiley Periodicals LLC on behalf of Botanical Society of America.)

This current study presents, for the first time, the complete chloroplast genome of two Cleomaceae species: Dipterygium glaucum and Cleome chrysantha in order to evaluate the evolutionary relationship. The cp genome is 158,576 bp in length with 35.74% GC content in D. glaucum and 158,111 bp with 35.96% GC in C. chrysantha. Inverted repeats IR 26,209 bp, 26,251 bp each, LSC of 87,738 bp, 87,184 bp and SSC of 18,420 bp, 18,425 bp respectively. There are 136 genes in the genome, which includes 80 protein coding genes, 31 tRNA genes and four rRNA genes were observed in both chloroplast genomes. 117 genes are unique while the remaining 19 genes are duplicated in IR regions. The analysis of repeats shows that the cp genome includes all types of repeats with more frequent occurrences of palindromic; Also, this analysis indicates that the total number of simple sequence repeats (SSR) were 323 in D. glaucum, and 313 in C. chrysantha, of which the majority of the SSRs in these plastid genomes were mononucleotide repeats A/T which are located in the intergenic spacer. Moreover, the comparative analysis of the four cp sequences revealed four hotspot genes (atpF, rpoC2, rps19, and ycf1), these variable regions could be used as molecular makers for the species authentication as well as resources for inferring phylogenetic relationships of the species. All the relationships in the phylogenetic tree are with high support, this indicate that the complete chloroplast genome is a useful data for inferring phylogenetic relationship within the Cleomaceae and other families. The simple sequence repeats identified will be useful for identification, genetic diversity, and other evolutionary studies of the species. This study reported the first cp genome of the genus Dipterygium and Cleome. The finding of this study will be beneficial for biological disciplines such as evolutionary and genetic diversity studies of the species within the core Cleomaceae.

Floral nectaries have evolved multiple times and rapidly diversified with the adaptive radiation of animal pollinators. As such, floral nectaries exhibit extraordinary variation in location, size, shape, and secretory mechanism. Despite the intricate ties to pollinator interactions, floral nectaries are often overlooked in morphological and developmental studies. As Cleomaceae exhibits substantial floral diversity, our objective was to describe and compare floral nectaries between and within genera. Floral nectary morphology was assessed through scanning electron microscopy and histology across three developmental stages of nine Cleomaceae species including representatives for seven genera. A modified fast green and safranin O staining protocol was used to yield vibrant sections without highly hazardous chemicals. Cleomaceae floral nectaries are most commonly receptacular, located between the perianth and stamens. The floral nectaries are supplied by vasculature, often contain nectary parenchyma, and have nectarostomata. Despite the shared location, components, and secretory mechanism, the floral nectaries display dramatic diversity in size and shape, ranging from adaxial protrusions or concavities to annular disks. Our data reveal substantive lability in form with both adaxial and annular floral nectaries interspersed across Cleomaceae. Floral nectaries contribute to the vast morphological diversity of Cleomaceae flowers and so are valuable for taxonomic descriptions. Though Cleomaceae floral nectaries are often derived from the receptacle and receptacular nectaries are common across flowering plants, the role of the receptacle in floral evolution and diversification is overlooked and warrants further exploration.

The autumn disappearance of Pontia chloridice from known sites in Cyprus suggests an ability of the species to remain in facultative pupal diapause during periods of unfavourable conditions, as is known with at least one other congeneric species. It is thought that P. chloridice is not migratory in the eastern Mediterranean, and that populations in Cyprus and NE Greece do not depend on reinforcement from immigration. [ABSTRACT FROM AUTHOR]

Cleomaceae is closely related to Brassicaceae and includes C3, C3–C4, and C4 species. Thus, this family represents an interesting system for studying the evolution of the carbon concentrating mechanism. However, inadequate genetic information on Cleomaceae limits their research applications. Here, we characterized 22 Cleomaceae accessions [3 genera (Cleoserrata , Gynandropsis , and Tarenaya) and 11 species] in terms of genome size; molecular phylogeny; as well as anatomical, biochemical, and photosynthetic traits. We clustered the species into seven groups based on genome size. Interestingly, despite clear differences in genome size (2C, ranging from 0.55 to 1.3 pg) in Tarenaya spp., this variation was not consistent with phylogenetic grouping based on the internal transcribed spacer (ITS) marker, suggesting the occurrence of multiple polyploidy events within this genus. Moreover, only G. gynandra , which possesses a large nuclear genome, exhibited the C4 metabolism. Among the C3-like species, we observed intra- and interspecific variation in nuclear genome size as well as in biochemical, physiological, and anatomical traits. Furthermore, the C3-like species had increased venation density and bundle sheath cell size, compared to C4 species, which likely predisposed the former lineages to C4 photosynthesis. Accordingly, our findings demonstrate the potential of Cleomaceae, mainly members of Tarenaya , in offering novel insights into the evolution of C4 photosynthesis. [ABSTRACT FROM AUTHOR]

Abstract Background The Capparaceae family is commonly recognized as a caper, while Cleomaceae represents one of small flowering family within the order Brassicales. Earlier, Cleomaceae was included in the family Capparaceae; then, it was moved to a distinct family after DNA evidence. Variation in habits and a bewildering array of floral and fruit forms contributed to making Capparaceae a “trash-basket” family in which many unrelated plants were placed. Indeed, family Capparaceae and Cleomaceae are in clear need of more detailed systematic revision. Results Here, in the present study, the morphological characteristics and the ecological distribution as well as the genetic diversity analysis among the twelve species of both Capparaceae and Cleomaceae have been determined. The genetic analysis has been checked using 15 ISSR, 30 SRAP, and 18 ISTR to assess the systematic knots between the two families. In order to detect the molecular phylogeny, a comparative analysis of the three markers was performed based on the exposure of discriminating capacity, efficiency, and phylogenetic heatmap. Our results indicated that there is a morphological and ecological variation between the two families. Moreover, the molecular analysis confirmed that ISTR followed by SRAP markers has superior discriminating capacity for describing the genetic diversity and is able to simultaneously distinguish many polymorphic markers per reaction. Indeed, both the PCA and HCA data have drawn a successful annotation relationship in Capparaceae and Cleome species to evaluate whether the specific group sort individual or overlap groups. Conclusion The outcomes of the morphological and ecological characterization along with the genetic diversity indicated an insight solution thorny interspecies in Cleome and Gynandropsis genera as a distinct family (Cleomaceae) and the other genera (Capparis, Cadaba, Boscia, and Maerua) as Capparaceae. Finally, we recommended further studies to elucidate the systematic position of Dipterygium glaucum.

Cleomaceae is closely related to Brassicaceae and includes C3, C3–C4, and C4 species. Thus, this family represents an interesting system for studying the evolution of the carbon concentrating mechanism. However, inadequate genetic information on Cleomaceae limits their research applications. Here, we characterized 22 Cleomaceae accessions [3 genera (Cleoserrata, Gynandropsis, and Tarenaya) and 11 species] in terms of genome size; molecular phylogeny; as well as anatomical, biochemical, and photosynthetic traits. We clustered the species into seven groups based on genome size. Interestingly, despite clear differences in genome size (2C, ranging from 0.55 to 1.3 pg) in Tarenaya spp., this variation was not consistent with phylogenetic grouping based on the internal transcribed spacer (ITS) marker, suggesting the occurrence of multiple polyploidy events within this genus. Moreover, only G. gynandra, which possesses a large nuclear genome, exhibited the C4 metabolism. Among the C3-like species, we observed intra- and interspecific variation in nuclear genome size as well as in biochemical, physiological, and anatomical traits. Furthermore, the C3-like species had increased venation density and bundle sheath cell size, compared to C4 species, which likely predisposed the former lineages to C4 photosynthesis. Accordingly, our findings demonstrate the potential of Cleomaceae, mainly members of Tarenaya, in offering novel insights into the evolution of C4 photosynthesis.

Premise Cleomaceae is emerging as a promising family to investigate a wide range of phenomena, such as C4 photosynthesis and floral diversity. However, functional techniques are lacking for elucidating this diversity. Herein, we establish virus‐induced gene silencing (VIGS) as a method of generating functional data for Cleome violacea, bolstering Cleomaceae as a model system. Methods We leveraged the sister relationship of Cleomaceae and Brassicaceae by using constructs readily available for Arabidopsis thaliana to provide initial information about the feasibility of VIGS in C. violacea. We then developed endogenous constructs to optimize VIGS efficiency and viability for fruit development. Results PHYTOENE DESATURASE was successfully downregulated in C. violacea using both heterologous and endogenous constructs. The endogenous construct had the highest degree of downregulation, with many plants displaying strong photobleaching. FRUITFULL‐treated plants were also successfully downregulated, with a high rate of survival but less effective silencing; only a small percentage of survivors showed a strong phenotype. Discussion Our optimized VIGS protocol in C. violacea enables functional gene analyses at different developmental stages. Additionally, C. violacea is amenable to heterologous knockdown, which suggests that a first pass using non‐endogenous constructs is a possible route to test additional species of Cleomaceae.

Premise: Cleomaceae is emerging as a promising family to investigate a wide range of phenomena, such as C4 photosynthesis and floral diversity. However, functional techniques are lacking for elucidating this diversity. Herein, we establish virus‐induced gene silencing (VIGS) as a method of generating functional data for Cleome violacea, bolstering Cleomaceae as a model system. Methods: We leveraged the sister relationship of Cleomaceae and Brassicaceae by using constructs readily available for Arabidopsis thaliana to provide initial information about the feasibility of VIGS in C. violacea. We then developed endogenous constructs to optimize VIGS efficiency and viability for fruit development. Results: PHYTOENE DESATURASE was successfully downregulated in C. violacea using both heterologous and endogenous constructs. The endogenous construct had the highest degree of downregulation, with many plants displaying strong photobleaching. FRUITFULL‐treated plants were also successfully downregulated, with a high rate of survival but less effective silencing; only a small percentage of survivors showed a strong phenotype. Discussion: Our optimized VIGS protocol in C. violacea enables functional gene analyses at different developmental stages. Additionally, C. violacea is amenable to heterologous knockdown, which suggests that a first pass using non‐endogenous constructs is a possible route to test additional species of Cleomaceae. [ABSTRACT FROM AUTHOR]

This current study presents, for the first time, the complete chloroplast genome of two Cleomaceae species: Dipterygium glaucum and Cleome chrysantha in order to evaluate the evolutionary relationship. The cp genome is 158,576 bp in length with 35.74% GC content in D. glaucum and 158,111 bp with 35.96% GC in C. chrysantha. Inverted repeats IR 26,209 bp, 26,251 bp each, LSC of 87,738 bp, 87,184 bp and SSC of 18,420 bp, 18,425 bp respectively. There are 136 genes in the genome, which includes 80 protein coding genes, 31 tRNA genes and four rRNA genes were observed in both chloroplast genomes. 117 genes are unique while the remaining 19 genes are duplicated in IR regions. The analysis of repeats shows that the cp genome includes all types of repeats with more frequent occurrences of palindromic; Also, this analysis indicates that the total number of simple sequence repeats (SSR) were 323 in D. glaucum , and 313 in C. chrysantha , of which the majority of the SSRs in these plastid genomes were mononucleotide repeats A/T which are located in the intergenic spacer. Moreover, the comparative analysis of the four cp sequences revealed four hotspot genes (atpF, rpoC2, rps19 , and ycf1), these variable regions could be used as molecular makers for the species authentication as well as resources for inferring phylogenetic relationships of the species. All the relationships in the phylogenetic tree are with high support, this indicate that the complete chloroplast genome is a useful data for inferring phylogenetic relationship within the Cleomaceae and other families. The simple sequence repeats identified will be useful for identification, genetic diversity, and other evolutionary studies of the species. This study reported the first cp genome of the genus Dipterygium and Cleome. The finding of this study will be beneficial for biological disciplines such as evolutionary and genetic diversity studies of the species within the core Cleomaceae. [ABSTRACT FROM AUTHOR]

Gynandropsis gynandra (L.) Briquet is a monoecious herb species in the family Cleomaceae. It is an important commercial crop with medicinal and nutritional values. Here, we sequenced, assembled, and analyzed the complete chloroplast genome of G. gynandra. The circular genome is 158,152 bp in size, consisting of two copies of inverted repeat (IR) regions of 26,181 bp, one large single-copy (LSC) region of 87,242 bp, and one small single-copy (SSC) region of 18,548 bp. The overall GC content was 35.81%. A total of 131 genes were annotated, including 37 tRNA genes, 87 protein-coding genes, and seven rRNA genes. Phylogenetic analysis based on 10 chloroplast genome sequences indicated that G. gynandra was more closely related to Tarenaya hassleriana.

In different lineages of C4 plants, the release of CO2 by decarboxylation of a C4 acid near rubisco is catalyzed by NADP-malic enzyme (ME) or NAD-ME, and the facultative use of phosphoenolpyruvate carboxykinase. The co-option of gene lineages during the evolution of C4-NADP-ME has been thoroughly investigated, whereas that of C4-NAD-ME has received less attention. In this work, we aimed at elucidating the mechanism of recruitment of NAD-ME for its function in the C4 pathway by focusing on the eudicot family Cleomaceae. We identified a duplication of NAD-ME in vascular plants that generated the two paralogs lineages: α- and β-NAD-ME. Both gene lineages were retained across seed plants, and their fixation was likely driven by a degenerative process of sub-functionalization, which resulted in a NAD-ME operating primarily as a heteromer of α- and β-subunits. We found most angiosperm genomes maintain a 1:1 β-NAD-ME/α-NAD-ME (β/α) relative gene dosage, but with some notable exceptions mainly due to additional duplications of β-NAD-ME subunits. For example, a significantly high proportion of species with C4-NAD-ME-type photosynthesis have a non-1:1 ratio of β/α. In the Brassicales, we found C4 species with a 2:1 ratio due to a β-NAD-ME duplication (β1 and β2); this was also observed in the C3Tarenaya hassleriana and Brassica crops. In the independently evolved C4 species, Gynandropsis gynandra and Cleome angustifolia, all three genes were affected by C4 evolution with α- and β1-NAD-ME driven by adaptive selection. In particular, the β1-NAD-MEs possess many differentially substituted amino acids compared with other species and the β2-NAD-MEs of the same species. Five of these amino acids are identically substituted in β1-NAD-ME of G. gynandra and C. angustifolia, two of them were identified as positively selected. Using synteny analysis, we established that β-NAD-ME duplications were derived from ancient polyploidy events and that α-NAD-ME is in a unique syntenic context in both Cleomaceae and Brassicaceae. We discuss our hypotheses for the evolution of NAD-ME and its recruitment for C4 photosynthesis. We propose that gene duplications provided the basis for the recruitment of NAD-ME in C4 Cleomaceae and that all members of the NAD-ME gene family have been adapted to fit the C4-biochemistry. Also, one of the β-NAD-ME gene copies was independently co-opted for its function in the C4 pathway.

In different lineages of C4 plants, the release of CO2 by decarboxylation of a C4 acid near rubisco is catalyzed by NADP-malic enzyme (ME) or NAD-ME, and the facultative use of phosphoenolpyruvate carboxykinase. The co-option of gene lineages during the evolution of C4-NADP-ME has been thoroughly investigated, whereas that of C4-NAD-ME has received less attention. In this work, we aimed at elucidating the mechanism of recruitment of NAD-ME for its function in the C4 pathway by focusing on the eudicot family Cleomaceae. We identified a duplication of NAD-ME in vascular plants that generated the two paralogs lineages: α - and β -NAD-ME. Both gene lineages were retained across seed plants, and their fixation was likely driven by a degenerative process of sub-functionalization, which resulted in a NAD-ME operating primarily as a heteromer of α- and β-subunits. We found most angiosperm genomes maintain a 1:1 β -NAD-ME /α -NAD-ME (β/α) relative gene dosage, but with some notable exceptions mainly due to additional duplications of β-NAD-ME subunits. For example, a significantly high proportion of species with C4-NAD-ME-type photosynthesis have a non-1:1 ratio of β/α. In the Brassicales, we found C4 species with a 2:1 ratio due to a β -NAD-ME duplication (β 1 and β 2); this was also observed in the C3 Tarenaya hassleriana and Brassica crops. In the independently evolved C4 species, Gynandropsis gynandra and Cleome angustifolia , all three genes were affected by C4 evolution with α- and β1- NAD-ME driven by adaptive selection. In particular, the β1-NAD-MEs possess many differentially substituted amino acids compared with other species and the β2-NAD-MEs of the same species. Five of these amino acids are identically substituted in β1-NAD-ME of G. gynandra and C. angustifolia , two of them were identified as positively selected. Using synteny analysis, we established that β -NAD-ME duplications were derived from ancient polyploidy events and that α -NAD-ME is in a unique syntenic context in both Cleomaceae and Brassicaceae. We discuss our hypotheses for the evolution of NAD-ME and its recruitment for C4 photosynthesis. We propose that gene duplications provided the basis for the recruitment of NAD-ME in C4 Cleomaceae and that all members of the NAD-ME gene family have been adapted to fit the C4-biochemistry. Also, one of the β -NAD-ME gene copies was independently co-opted for its function in the C4 pathway. [ABSTRACT FROM AUTHOR]

Premise: Whole‐genome duplications (WGDs) are prevalent throughout the evolutionary history of plants. For example, dozens of WGDs have been phylogenetically localized across the order Brassicales, specifically, within the family Brassicaceae. A WGD event has also been identified in the Cleomaceae, the sister family to Brassicaceae, yet its placement, as well as that of WGDs in other families in the order, remains unclear. Methods: Phylo‐transcriptomic data were generated and used to infer a nuclear phylogeny for 74 Brassicales taxa. Genome survey sequencing was also performed on 66 of those taxa to infer a chloroplast phylogeny. These phylogenies were used to assess and confirm relationships among the major families of the Brassicales and within Brassicaceae. Multiple WGD inference methods were then used to assess the placement of WGDs on the nuclear phylogeny. Results: Well‐supported chloroplast and nuclear phylogenies for the Brassicales and the putative placement of the Cleomaceae‐specific WGD event Th‐ɑ are presented. This work also provides evidence for previously hypothesized WGDs, including a well‐supported event shared by at least two members of the Resedaceae family, and a possible event within the Capparaceae. Conclusions: Phylogenetics and the placement of WGDs within highly polyploid lineages continues to be a major challenge. This study adds to the conversation on WGD inference difficulties by demonstrating that sampling is especially important for WGD identification and phylogenetic placement. Given its economic importance and genomic resources, the Brassicales continues to be an ideal group for assessing WGD inference methods. [ABSTRACT FROM AUTHOR]

Tarenaya minutiflora Iltis & Cochrane is described from Guyana. The new species, distinguished from its congeners by the very small size of the flowers, is illustrated and compared to its probable nearest relatives T. parviflora (Kunth) Iltis, T. psoraleifolia (DC.) Soares Neto & Roalson, and T. brasiliensis Weinm. [ABSTRACT FROM AUTHOR]

Species in the genera Hyaloperonospora and Peronospora (Oomycota, Peronosporaceae) are obligate biotrophic pathogens responsible for destructive downy mildew diseases of ornamental horticulture plants worldwide. Throughout the past decade, newly emergent downy mildew diseases have become increasingly prevalent, but often the identity of the pathogen is unknown or poorly defined. In this study we set out to identify the downy mildew pathogens infecting three widely grown ornamental plants: Agastache spp. (hyssops), Tarenaya hassleriana (syn. = Cleome hassleriana; spider-flower) and Monarda didyma (bee balm or bergamot). Phylogenetic analysis of the internal transcribed spacer rDNA and cox2 revealed two new species, which are described here as Peronospora monardae infecting both Monarda didyma and A. mexicana, and Hyaloperonospora daughtreyae infecting T. hassleriana. One sample of Peronospora collected from Agastache sp. 'Bolero' did not cluster with P. monardae, but its relationship to other species in the clade was not sufficiently resolved for taxonomic determination. Phylogenetic analysis also supported the combination of Peronospora iberidis, a pathogen of Iberis spp. (candytuft), into Hyaloperonospora. To assess intrasample ITS sequence variation from P. monardae, data were generated using high-throughput amplicon sequencing (HTAS). Multiple haplotypes were identified from the P. monardae samples, with less than 1% intrasample variation observed. From a core set of haplotypes shared across the HTAS the variant frequencies could not be completely explained by heterozygosity alone. [ABSTRACT FROM AUTHOR]

The family Cleomaceae is represented in the Neotropics by nine native lineages, with some of them currently recognized as distinct genera and others in need of clear delimitation. We present here a new phylogenetic hypothesis for the New World Cleomaceae with a more comprehensive sampling, including samples of 62 species with emphasis on Tarenaya and based on nrDNA ITS and cpDNA matK and ndhF. Six clades are supported: (1) a new clade comprising Cleome tenuis and Physostemon humilis; and five other previously identified clades: (2) the Dactylaena clade, including the monophyletic Dactylaena and the paraphyletic Physostemon and Mitostylis; (3) the Andean clade, comprising Andinocleome and Podandrogyne; (4) the Melidiscus clade; (5) the Cleoserrata clade, and (6) the Tarenaya clade; the last three corresponding to genera with the same name. The lineage comprising C. tenuis and P. humilis is strongly supported by morphological and molecular datasets, and a new genus, named Iltisiella, comprising these species is proposed here. Tarenaya is clearly demonstrated to be a monophyletic genus, corroborating previous analyses. We identify four subclades in its topology, and the relationships among the species in each subclade revealed to be moderately to strongly resolved. Furthermore, we conducted a historical biogeographical analysis of the New World Cleomaceae on time‐calibrated trees, and our results suggest an Oligocene origin at least 35.02 Mya with a MRCA distributed in Mesoamerica and the Northern Andes. For Tarenaya, our results suggest a Miocene origin at least 17 Mya, with the most recent common ancestor in the Chacoan and Atlantic Forest geographic areas. [ABSTRACT FROM AUTHOR]

Background: The Capparaceae family is commonly recognized as a caper, while Cleomaceae represents one of small flowering family within the order Brassicales. Earlier, Cleomaceae was included in the family Capparaceae; then, it was moved to a distinct family after DNA evidence. Variation in habits and a bewildering array of floral and fruit forms contributed to making Capparaceae a “trash-basket” family in which many unrelated plants were placed. Indeed, family Capparaceae and Cleomaceae are in clear need of more detailed systematic revision. Results: Here, in the present study, the morphological characteristics and the ecological distribution as well as the genetic diversity analysis among the twelve species of both Capparaceae and Cleomaceae have been determined. The genetic analysis has been checked using 15 ISSR, 30 SRAP, and 18 ISTR to assess the systematic knots between the two families. In order to detect the molecular phylogeny, a comparative analysis of the three markers was performed based on the exposure of discriminating capacity, efficiency, and phylogenetic heatmap. Our results indicated that there is a morphological and ecological variation between the two families. Moreover, the molecular analysis confirmed that ISTR followed by SRAP markers has superior discriminating capacity for describing the genetic diversity and is able to simultaneously distinguish many polymorphic markers per reaction. Indeed, both the PCA and HCA data have drawn a successful annotation relationship in Capparaceae and Cleome species to evaluate whether the specific group sort individual or overlap groups. Conclusion: The outcomes of the morphological and ecological characterization along with the genetic diversity indicated an insight solution thorny interspecies in Cleome and Gynandropsis genera as a distinct family (Cleomaceae) and the other genera (Capparis, Cadaba, Boscia, and Maerua) as Capparaceae. Finally, we recommended further studies to elucidate the systematic position of Dipterygium glaucum. [ABSTRACT FROM AUTHOR]

The Cleomaceae Bercht. and J. Presl family consists of seven genera and 35 species found across Brazil. Studies have registered the use of species in traditional medicine and for wider purposes. Within this context, this review aims to provide updated, comprehensive, and categorised information on botanical, ethnobotanical, chemical, and pharmacological studies of Cleomaceae species in Brazil to identify their therapeutic potential and prompt future research opportunities. To this end, an integrative review analysing articles from different databases using different combinations of keywords in Portuguese and English was carried out. The search strategy was adapted to the selected databases following each of their search criteria. The Boolean operators 'AND' and 'OR' were used to combine the terms. Articles published between 2001 and 2022 were included. Around 90.9% of ethnobotanical studies found were carried out in the Northeast region. A total of eight plant species were identified, distributed among four genera. The category of medicinal use was the most frequent (36.37%), and the roots were part of the plants used the most (30.44%). It was observed that most species are used to treat respiratory diseases. Few botanical studies and chemical and pharmacological studies were conducted, showing a lack of research in these areas. In addition, the number of species explored in studies demonstrates the need for investment in the identification of species within the family, as well as in the discovery of new bioactive compounds from species of Cleomaceae in Brazil and their pharmacological applications. [Display omitted] • Brazil is considered the country with the greatest plant diversity on the planet. • Species of the Cleomaceae family in Brazil have different traditional uses. • In Brazil, the relevance of Cleomaceae species has not been scientifically defined. [ABSTRACT FROM AUTHOR]

In this study, we present the complete chloroplast genome of Dipterygium glaucum an important medicinal plant in Saudi Arabia, in order to evaluate the evolutionary relationship in the family Cleomaceae. The cp genome is second to be sequenced in Cleomaceae family, it has a circular structure and a length of 158,576 bp with 35.74% GC content. The genome is divided into four regions: LSC of 87,738 bp, SSC of 18,420 bp and a pair of inverted repeats 26,209 bp each. The chloroplast genome of D. glaucum encodes 116 different genes, including 81protien coding genes, 31tRNA genes and four rRNA genes. The phylogenetic relationship showed a close relationship between D. glaucum and Tarenaia hassleriana.

Background and Aims Bat-pollination is an important system in terms of occurrence and distribution, although it remains little studied. Thus, the role of particular flower traits in this interaction remains uncertain. Flower height along the shoot axis, associated with flower exposure, has often been deemed a key trait in this system, but its effect on fitness has not previously been assessed. We aimed to test its role and propose that taller flowers attain higher fitness due to a higher degree of accessibility and conspicuity to foraging bats. Methods We assessed the effect of floral height on bat visiting rates to individual flowers of Crescentia cujete (Bignoniaceae), a cauliflorous model bat-pollinated species with a marked gradient in flower height along the shoot axis. Additionally, we tested the effect of this variable on seed/ovule ratio measurements from seven other species from different families along a herb–tree gradient. Hypotheses were tested through mixed-effect linear models. Key Results Bat visiting rates varied positively as a function of flower height in C. cujete , but significance was found only for the subset of flowers located on the trunk, closer to the ground. Similarly, seed/ovule ratios were positively correlated with flower height only for the three species with the shortest statures along the height gradient and shortest average floral heights. These results suggest that proximity to the ground, associated with herbaceous or bushy surrounding vegetation, may be an obstacle to the foraging of nectar-feeding bats, which in turn should explain the morphological convergence of inflorescence length and exposure strategies of short-statured bat-pollinated plants. Conclusions Flower height has a species-specific effect on plant fitness. This study provides a novel numerical perspective to the role of an unexplored trait in bat-pollination, and has elucidated some aspects of the adaptive importance of flower height based on limitations imposed by ecologically complex pollinators. [ABSTRACT FROM AUTHOR]

A new species of the genus Cleome L. from Pakistan is described and illustrated. The new species is described under the name C. karachiensis sp. nov. and compared with two closely related species i.e., Cleome brachycarpa and C. viscosa in terms of morphology, palynology, seed morphology and seed protein profile. A key to the species of genus Cleome L. from Pakistan is also provided. [ABSTRACT FROM AUTHOR]

Background: Floral organs are specified by MADS-domain transcription factors that act in a combinatorial manner, as summarized in the (A)BCE model. However, this evolutionarily conserved model is in contrast to a remarkable amount of morphological diversity in flowers. One of the mechanisms suggested to contribute to this diversity is duplication of floral MADS-domain transcription factors. Although gene duplication is often followed by loss of one of the copies, sometimes both copies are retained. If both copies are retained they will initially be redundant, providing freedom for one of the paralogs to change function. Here, we examine the evolutionary fate and functional consequences of a transposition event at the base of the Brassicales that resulted in the duplication of the floral regulator PISTILLATA (PI), using Tarenaya hassleriana (Cleomaceae) as a model system. Results: The transposition of a genomic region containing a PI gene led to two paralogs which are located at different positions in the genome. The original PI copy is syntenic in position with most angiosperms, whereas the transposed copy is syntenic with the PI genes in Brassicaceae. The two PI paralogs of T. hassleriana have very similar expression patterns. However, they may have diverged in function, as only one of these PI proteins was able to act heterologously in the first whorl of A. thaliana flowers. We also observed differences in protein complex formation between the two paralogs, and the two paralogs exhibit subtle differences in DNA-binding specificity. Sequence analysis indicates that most of the protein sequence divergence between the two T. hassleriana paralogs emerged in a common ancestor of the Cleomaceae and the Brassicaceae. Conclusions: We found that the PI paralogs in T. hassleriana have similar expression patterns, but may have diverged at the level of protein function. Data suggest that most protein sequence divergence occurred rapidly, prior to the origin of the Brassicaceae and Cleomaceae. It is tempting to speculate that the interaction specificities of the Brassicaceae-specific PI proteins are different compared to the PI found in other angiosperms. This could lead to PI regulating partly different genes in the Brassicaceae, and ultimately might result in change floral in morphology. [ABSTRACT FROM AUTHOR]

We wish to report the study of the antimicrobial activity of plants collected in the vicinity of a geothermal area in Perak, Malaysia. This is the first report of these species in the vicinity of thextis geothermal area. The plants are Cleome icosandra, locally known as Maman Pasir of the family Cleomaceae, and Stachytarpheta species of the family Lamiaceae. Both are subshrubs and are believed to have specific biological activities as a result of living in such extreme areas. Methanol extracts of both plants revealed no antimicrobial activity against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa.

Photosynthesis in different organs of Cleome was analysed in four species known to have differences in leaf photosynthesis: Cleome africana Botsch. (C3), Cleome paradoxa R.Br. (C3-C4 intermediate), Cleome angustifolia Forssk. and Cleome gynandra L. (C4). The chlorophyll content, carbon isotope composition, stomatal densities, anatomy, levels and compartmentation of some key photosynthetic enzymes, and the form and function of photosynthesis were determined in different organs of these species. In the three xerophytes, C. africana , C. paradoxa , and C. angustifolia , multiple organs contribute to photosynthesis (cotyledons, leaves, petioles, stems and pods) which is considered important for their survival under arid conditions. In C. africana , all photosynthetic organs have C3 photosynthesis. In C. paradoxa , cotyledons, leaves, stems and petioles have C3-C4 type features. In C. angustifolia , the pods have C3 photosynthesis, whereas all other organs have C4 photosynthesis with Kranz anatomy formed by a continuous, dual layer of chlorenchyma cells. In the subtropical C4 species C. gynandra , cotyledons, leaves, and pods develop C4 photosynthesis, with Kranz anatomy around individual veins; but not in stems and petioles which have limited function of photosynthesis. The diversity in forms and the capacity of photosynthesis in organs of these species to contribute to their carbon economy is discussed. There is great interest in plants that capture CO2 from the atmosphere and concentrate it in leaves to support photosynthesis. The present study shows the diversity in expression and function of this and other forms of photosynthesis in cotyledons, leaves, stems, petioles and pods during the life cycle of Cleome species. Further investigation into the genetic regulation of development of these forms within and between species is needed. [ABSTRACT FROM AUTHOR]

Cleome rutidosperma DC, commonly known in Jamaica as ‘consumption-weed’ is a plant traditionally used in folklore for treating tuberculosis and other infectious and chronic ailments. We evaluate for the first time the chemical composition and biological activities of the essential oil components of the complete aerial parts of this plant. The essential oil obtained by steam distillation (0.02%) was analyzed by a combination of gas chromatography-flame ionization detector (GC-FID), gas chromatography-mass spectroscopy (GC–MS) and retention index (RI). The volatile oil of C. rutidosperma was dominated by oxygenated diterpenes (67.6%); with ( Z )-phytol (65.1%) being the single most abundant constituent. C. rutidosperma aerial essential oil exhibited moderate inhibition against the activity of recombinant arylamine N-acetyltransferase (NAT) from Mycobacterium marinum (IC 50 22.20 ± 1.80 μg/μL), while, racemic phytol had an inhibition with an IC 50 of 22.33 μg/μL ± 0.50 μg/μL, thus accounting for the NAT inhibition imparted by the crude oil. Inhibition of NAT, a key enzyme in mycobacterial growth may be the pathway in which phytol, shown in this study to interact with the active site using in-silico methods, renders its previously demonstrated anti-tubercular properties. The phytol rich essential oil also demonstrated antimicrobial activity against all nine human pathogenic bacteria and the fungus strain assayed, with the most significant inhibitory activity against Bacillus cereus and justifies the need for further evaluation and development of the essential oils from this plant. [ABSTRACT FROM AUTHOR]

Cleomaceae is a diverse group well-suited to addressing fundamental genomic and evolutionary questions as the sister group to Brassicaceae, facilitating transfer of knowledge from the model Arabidopsis thaliana . Phylogenetic and taxonomic revisions provide a framework for examining the evolution of substantive morphological and physiology diversity in Cleomaceae, but not necessarily in Brassicaceae. The investigation of both nested and contrasting whole-genome duplications (WGDs) between Cleomaceae and Brassicaceae allows comparisons of independently duplicated genes and investigation of whether they may be drivers of the observed innovations. Further, a wealth of outstanding genetic research has provided insight into how the important alternative carbon fixation pathway, C 4 photosynthesis, has evolved via differential expression of a suite of genes, of which the underlying mechanisms are being elucidated. [ABSTRACT FROM AUTHOR]

Summary: Unlike most ancient microRNAs, which conservatively target homologous genes across species, microRNA827 (miR827) targets two different types of SPX (SYG1/PHO81/XPR1)‐domain‐containing genes, NITROGEN LIMITATION ADAPTATION (NLA) and PHOSPHATE TRANSPORTER 5 (PHT5), in Arabidopsis thaliana and Oryza sativa to regulate phosphate (Pi) transport and storage, respectively. However, how miR827 shifted its target preference and its evolutionary history are unknown. Based on target prediction analysis, we found that in most angiosperms, miR827 conservatively targets PHT5 homologs, but in Brassicaceae and Cleomaceae it preferentially targets NLA homologs, and we provide evidence for the transition of target preference during Brassicales evolution. Intriguingly, we found a lineage‐specific loss of the miR827‐regulatory module in legumes. Analysis of miR827‐mediated cleavage efficiency and the expression of PHT5 in A. thaliana indicated that accumulation of mutations in the target site and the exclusion of the target site by alternative transcriptional initiation eliminated PHT5 targeting by miR827. Here, we identified a transition of miR827 target preference during plant evolution and revealed the uniqueness of miR827‐mediated regulation among conserved plant miRNAs. Despite the change in its target preference, upregulation of miR827 by Pi starvation and its role in regulating cellular Pi homeostasis were retained. [ABSTRACT FROM AUTHOR]