Impact of the climatic changes in the Pliocene-Pleistocene transition on Irano-Turanian species. The radiation of genus Jurinea (Compositae).

The Irano-Turanian region is one of the world's richest floristic regions and the centre of diversity for numerous xerophytic plant lineages. However, we still have limited knowledge on the timing of evolution and biogeographic history of its flora, and potential drivers of diversification remain underexplored. To fill this knowledge gap, we focus on the Eurasian genus Jurinea (ca. 200 species), one of the largest plant radiations that diversified in the region. We applied a macroevolutionary integrative approach to explicitly test diversification hypotheses and investigate the relative roles of geography vs. ecology and niche conservatism vs. niche lability in speciation processes. To do so, we gathered a sample comprising 77% of total genus richness and obtained data about (1) its phylogenetic history, recovering 502 nuclear loci sequences; (2) growth forms; (3) ecological niche, compiling data of 21 variables for more than 2500 occurrences; and (4) paleoclimatic conditions, to estimate climatic stability. Our results revealed that climate was a key factor in the evolutionary dynamics of Jurinea. The main diversification and biogeographic events that occurred during past climate changes, which led to colder and drier conditions, are the following: (1) the origin of the genus (10.7 Ma); (2) long-distance dispersals from the Iranian Plateau to adjacent regions (∼7-4 Ma); and (3) the diversification shift during Pliocene-Pleistocene Transition (ca. 3 Ma), when net diversification rate almost doubled. Our results supported the pre-adaptation hypothesis, i.e., the evolutionary success of Jurinea was linked to the retention of the ancestral niche adapted to aridity. Interestingly, the paleoclimatic analyses revealed that in the Iranian Plateau long-term climatic stability favoured old-lineage persistence, resulting in current high species richness of semi-arid and cold adapted clades; whereas moderate climate oscillations stimulated allopatric diversification in the lineages distributed in the Circumboreal region. In contrast, growth form lability and high niche disparity among closely related species in the Central Asian clade suggest adaptive radiation to mountain habitats. In sum, the radiation of Jurinea is the result of both adaptive and non-adaptive processes influenced by climatic, orogenic and ecological factors.

African Mountain Thistles: Three New Genera in the Carduus-Cirsium Group.

The floras on the highest mountains in tropical eastern Africa are among the most unique floras in the world. Despite the exceptionally high concentration of endemic species, these floras remain understudied from an evolutionary point of view. In this study, we focus on the Carduus-Cirsium group (subtribe Carduinae) to unravel the evolutionary relationships of the species endemic to the tropical Afromontane and Afroalpine floras, aiming to improve the systematics of the group. We applied the Hyb-Seq approach using the Compositae1061 probe set on 190 samples (159 species), encompassing representatives of all genera of Carduinae. We used two recently developed pipelines that enabled the processing of raw sequence reads, identification of paralogous sequences and segregation into orthologous alignments. After the implementation of a missing data filter, we retained sequences from 986 nuclear loci and 177 plastid regions. Phylogenomic analyses were conducted using both concatenated and summary-coalescence methods. The resulting phylogenies were highly resolved and revealed three distinct evolutionary lineages consisting of the African species traditionally referred to as Carduus and Cirsium. Consequently, we propose the three new genera Afrocarduus, Afrocirsium and Nuriaea; the latter did notably not belong to the Carduus-Cirsium group. We detected some incongruences between the phylogenies based on concatenation vs. coalescence and on nuclear vs. plastid datasets, likely attributable to incomplete lineage sorting and/or hybridization.

Synergetic effect of water deficit and arbuscular mycorrhizal symbiosis on the expression of aquaporins in wheat (Triticum aestivum L.) roots: insights from NGS RNA-sequencing.

Wheat (Triticum aestivum) is one of the most important crops in the world. This investigation was attempted to evaluate the transcriptional responses of aquaporins (AQPs) to the mycorrhizal inoculation and/or water deficit conditions in wheat to clarify how the arbuscular mycorrhizal symbiosis can contribute to the modulation of water homeostasis. The wheat seedlings were subjected to the water deficiency, and mycorrhizal inoculation using arbuscular fungus Funneliformis mosseae and Illumina RNA-Seq analyses confirmed that aquaporins expressed differentially in response to both the irrigation levels and mycorrhizal colonization. Results of this study showed that only 13% of the studied AQPs were responsive to water deficit with a tiny fraction (3%) being up-regulated. Mycorrhizal inoculation had a greater impact on the expression of AQPs with ca. 26% being responsive, ca. 4% of which were up-regulated. The samples with arbuscular mycorrhizal inoculation yielded more root and stem biomass. Water deficit and mycorrhizal inoculation caused different AQPs to be up-regulated. The effect of mycorrhizal inoculation on the expression of AQPs was intensified by applying water deficiency with 32% of studied AQPs being responsive, 6% of which up-regulated. We also found that the overexpression of three genes TaNIP1-10, TaNIP3-3, and TaNIP3-4 was chiefly triggered by mycorrhizal inoculation. Our results show that water deficit has a lower impact on the expression of aquaporins compared to what the arbuscular mycorrhizal inoculation has; water deficit and arbuscular mycorrhizal inoculation mainly cause the down-regulation of the aquaporins, and water deficit and the arbuscular inoculation have synergetic effects. These findings could improve our knowledge of how arbuscular mycorrhizal symbiosis can contribute to the modulation of water homeostasis. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01285-w.

Phylogenomics and morphological evolution of the mega-diverse genus Artemisia (Asteraceae: Anthemideae): implications for its circumscription and infrageneric taxonomy.

BACKGROUND AND AIMS: Artemisia is a mega-diverse genus consisting of ~400 species. Despite its medicinal importance and ecological significance, a well-resolved phylogeny for global Artemisia, a natural generic delimitation and infrageneric taxonomy remain missing, owing to the obstructions from limited taxon sampling and insufficient information on DNA markers. Its morphological characters, such as capitulum, life form and leaf, show marked variations and are widely used in its infrageneric taxonomy. However, their evolution within Artemisia is poorly understood. Here, we aimed to reconstruct a well-resolved phylogeny for global Artemisia via a phylogenomic approach, to infer the evolutionary patterns of its key morphological characters and to update its circumscription and infrageneric taxonomy. METHODS: We sampled 228 species (258 samples) of Artemisia and its allies from both fresh and herbarium collections, covering all the subgenera and its main geographical areas, and conducted a phylogenomic analysis based on nuclear single nucleotide polymorphisms (SNPs) obtained from genome skimming data. Based on the phylogenetic framework, we inferred the possible evolutionary patterns of six key morphological characters widely used in its previous taxonomy. KEY RESULTS: The genus Kaschgaria was revealed to be nested in Artemisia with strong support. A well-resolved phylogeny of Artemisia consisting of eight highly supported clades was recovered, two of which were identified for the first time. Most of the previously recognized subgenera were not supported as monophyletic. Evolutionary inferences based on the six morphological characters showed that different states of these characters originated independently more than once. CONCLUSIONS: The circumscription of Artemisia is enlarged to include the genus Kaschgaria. The morphological characters traditionally used for the infrageneric taxonomy of Artemisia do not match the new phylogenetic tree. They experienced a more complex evolutionary history than previously thought. We propose a revised infrageneric taxonomy of the newly circumscribed Artemisia, with eight recognized subgenera to accommodate the new results.

Chemical compositions of the essential oils from Iranian populations of Rhabdosciadium aucheri Boiss. (Apiaceae).

Rhabdosciadium aucheri has a limited distribution in west, center, and south of Iran. The aim of the present study was to assess essential oil content and variability in seven natural populations employing GC-FID and GC-MS analyses. Aerial parts of the specimens were collected at the full flowering stage. Overall, 27 chemical components were determined in the populations. The essential oil yields varied from 0.05% to 0.12%. The most abundant compounds were germacrene D (10.7-51.7%), ß-sesquiphellandrene (0.8-54.6%), ß-elemene (0.3-39.5%), juniper camphor (3.5-20.8%), limonene (0.4-19.6%) and α-pinene (0.4-4.5%) among the populations. Due to some variations observed in the essential oil components between populations, four different chemotypes were recognized including Chemotype I (germacrene D/ß-sesquiphellandrene/juniper camphor), Chemotype II (ß-elemene/germacrene D), Chemotype III (germacrene D/ß-sesquiphellandrene) and Chemotype IV (ß-elemene/germacrene D). The variations that occurred in chemical compositions among populations could be valuable in the pharmaceutical industries and conservation strategies.

Genetic structure of Alhagi (Hedysareae, Fabaceae) populations using ISSR data in Iran.

Alhagi Gagnebin (Fabaceae: Hedysareae) is a small genus of shrubs or subshrubs distributed in temperate and tropical regions of Asia, Europe, and Africa. The exact number of Alhagi species in Iran has been disputable. Studies showed that morphology, seed protein, and chromosome characters could not clearly delimitate the species boundaries of the genus. In order to determine species boundaries and the population structure of Alhagi species in Iran, eight labeled inter simple sequence repeat (ISSR) primers were used to screen 22 populations including 110 individuals. STRUCTURE, PCoA and clustering analyses of ISSR data were able to clearly split all populations of Alhagi in Iran into two distinct groups. As the populations A. pseudalhagi and A. maurorum jointly formed a group (A. maurorum complex) segregated from those of Alhagi graecorum. Further analyses of A. maurorum complex showed a significant molecular difference among the studied populations (PhiPT value = 0.213, P = 0.001) as well as a high amount of variability within populations (79%) indicating frequent genetic exchanges. Structure analysis of complex populations (K = 2) showed no distinct genetic pattern related to their geographical distribution. In this study, Alhagi has retrieved the only two species of Alhagi in Iran; A. graecorum and A. maurorum.

Antibacterial effect of carbon nanotube containing chemical compounds on drug-resistant isolates of Acinetobacter baumannii.

BACKGROUND AND OBJECTIVES: Acinetobacter baumannii is recognized as an important pathogen responsible for serious infections causing episodes of hospital infection. Carbon nanotubes (CNTs) have recently emerged as superior materials against antibiotic-resistant bacteria. In this study, a new chemical compound was designed in order to combat A. baumannii infections. Subsequently, the effect of this novel carbon nanotube coated with an antibacterial compound on Extensively Drug-Resistant (XDR), Multidrug-Resistant (MDR) and Pan-Drug-Resistance (PDR) strains of A. baumannii was investigated. MATERIALS AND METHODS: A total of 122 clinical isolates of A. baumannii were cultured from burn patients and their susceptibility to antibiotics were checked using disk diffusion method and Minimum inhibitory concentration. Antimicrobial effects of the coated carbon nanotube were evaluated on XDR, MDR and PDR isolates of A. baumannii. Cell viability was determined using tetrazolium reduction assay (MTT) on human fibroblast cell line (HDFa). Wound healing processes were assessed by quantitative polymerase chain reaction. RESULTS: Of the 50 A. baumannii isolates, 38 (76%) were found to be MDR and 12 (24%) were XDR. No PDR strains were detected. Results indicated that the carbon nanotube combined with mercury had antibacterial effect against different A. baumannii species and it also was able to increase the expression of epidermal growth factor, platelet-derived growth factor and vascular endothelial growth factor A mRNA levels which are involved in wound healing. CONCLUSION: The engineered carbon nanotube compound can potentially be used for treatment of burn related infections. This can potentially give clinicians a new tool for treating A. baumannii infections.

RAPD-based evaluation of genetic diversity among populations of the Iranian endemic species Rhabdosciadium aucheri Boiss. (Apiaceae).

Rhabosciadium aucheri is an Iranian endemic herbaceous species that grows in the west, center, and south regions of Iran. In the present study, genetic variation of 70 individuals belonging to seven natural populations of four provinces was investigated using Random Amplified Polymorphic DNA (RAPD) markers. Ten out of twenty-two RAPD primers employed in this study, generated 110 highly amplified and reproducible loci and a mean of 11.1 bands per primer and 48.13% of polymorphism was obtained. According to our results, the primer OPA10 presented the highest effective number of alleles, Shannon's index, and genetic diversity. The highest value of genetic identity (0.916) was determined between Hamadan, Nahavand and Hamadan, Alvand Mts. populations and the highest genetic distance (0.277) was observed between Hamadan, Asadabad and Kurdistan, Qorveh populations. Therefore, there is an obvious correlation between genetic diversity and geographical distribution. PCA was obtained based on RAPD molecular data and Neighbor Joining (NJ) dendrogram was provided successively. Similar results were attained employing UPGMA and Neighbor Joining dendrograms, supported by PCA ordination plot. Overall, almost moderate level of polymorphism was obtained by RAPD molecular markers at the population level that shows a reasonable amount of intraspecific variability. The reason might be due to the low level of gene flow between populations that could give rise to high genetic differentiation.

The Construction of Carbon Nanotubes Containing an Anti-Bacterial Chemical Component and its Effect on MDR and XDR Isolates of Pseudomonas Aeruginosa.

BACKGROUND: Pseudomonas aeruginosa is an opportunistic human pathogen that causes severe acute and chronic nosocomial infections, especially in immunocompromised burn patients. and can lead to severe mortality and morbidity. The emergence of antibiotic resistant P. aeruginosa infections has created significant challenges in treating these patients. A potential alternative treatment for antibiotic resistant pathogens includes the use of carbon nanotubes (CNTs), which have received considerable attention due to their potent antibacterial activity. The aim of this study was to construct a novel CNT containing an anti-bacterial chemical component to effectively combat drug resistant P. aeruginosa infections. METHODS: In this study, a novel chemical component was synthesized and coated the CNT. The antimicrobial effects were then evaluated on MDR, XDR, and PDR strains of P. aeruginosa isolated from burn patients. Antibiotic susceptibility was evaluated using the disk diffusion test and minimum inhibitory concentration (MIC) testing. In order to determine the potential cytotoxicity, an MTT assay was performed on Human Dermal Fibroblasts. The effect of treatment on the expression of wound healing genes was analyzed via qRT-PCR. RESULTS: Experimental data indicates that our CNT coated chemical compound had antibacterial properties, negligible cytotoxicity, and could accelerate the wound healing process. CONCLUSION: Given the antibacterial properties of our CNT chemical compound, it has the potential to treat and reduce the occurrence of multi-drug resistant P. aeruginosa burn wound infections and aid in wound healing by turning on genes (VEGFA, EGF and PDEGF) involved in the wound healing process.

Impact of arbuscular mycorrhizal fungi (AMF) on gene expression of some cell wall and membrane elements of wheat (Triticum aestivum L.) under water deficit using transcriptome analysis.

Mycorrhizal symbiotic relationship is one of the most common collaborations between plant roots and the arbuscular mycorrhizal fungi (AMF). The first barrier for establishing this symbiosis is plant cell wall which strongly provides protection against biotic and abiotic stresses. The aim of this study was to investigate the gene expression changes in cell wall of wheat root cv. Chamran after inoculation with AMF, Funneliformis mosseae under two different irrigation regimes. To carry out this investigation, total RNA was extracted from the roots of mycorrhizal and non-mycorrhizal plants, and analyzed using RNA-Seq in an Illumina Next-Seq 500 platform. The results showed that symbiotic association between wheat and AMF and irrigation not only affect transcription profile of the plant growth, but also cell wall and membrane components. Of the 114428 genes expressed in wheat roots, the most differentially expressed genes were related to symbiotic plants under water stress. The most differentially expressed genes were observed in carbohydrate metabolic process, lipid metabolic process, cellulose synthase activity, membrane transports, nitrogen compound metabolic process and chitinase activity related genes. Our results indicated alteration in cell wall and membrane composition due to mycorrhization and irrigation regimes might have a noteworthy effect on the plant tolerance to water deficit.

Development of an immunoblotting assay for serodiagnosis of Burkholderia mallei infection: the whole-cell proteome-based paradigm.

BACKGROUND AND OBJECTIVES: Burkholderia mallei is the leading cause of glanders, a highly transmittable and an OIE-notifiable disease of equidae. Despite the importance of B. mallei, little is known about serodiagnosis of glanders. The present study aimed to develop an immunoblotting assay based on whole-cell proteome of B. mallei to enable accurate serodiagnosis of glanders. MATERIALS AND METHODS: Three farm horses were subcutaneously immunized with a crude suspension (106 cfu/ml) of heat-inactivated B. mallei formulated with incomplete Freund's adjuvant (IFA) to achieve a hyperimmune sera panel. The immunization was done for 1, 14 and 28 days with 1 dose of 1 ml antigen containing 106 cfu/ml. The hyperimmunity of sera was confirmed by CFT. B. mallei whole-cell proteome was prepared through sonication and the protein content was visualized by SDS-PAGE and quantified by Western blot using HRP-conjugated rabbit anti-horse IgG. A comprehensive set of positive and negative horse sera validated the test. RESULTS: A ladder pattern of the B. mallei immunoreactive antigens was seen within the region of 20-90 kDa clearly and the immunoblot was scored positive, while no reaction was seen for the negative sera. The Western blot assay indicated a noticeably higher diagnostic specificity for positive or negative sera of glanders. CONCLUSION: The whole-cell proteome-based immunoblot proved reliable and straightforward in our study. The prepared antigen was adaptable for application in immunoblotting. We assumed this improved immunoblotting system provides appropriate sensitivity and also specificity expected in serodiagnosis of glanders in endemic areas and typically in less-developed countries.

Diarylbutyrolactone lignans from Linum corymbulosum in vitro cultures.

The arylbutyrolactone lignans (-)-hinokinin, 3,4 : 3',4'-bis(methylenedioxy)-lign-7( E)-en-9,9'-olide and 3,4 : 3',4'-bis(methylenedioxy)-lign-7( Z)-en-9,9'-olide, hitherto unknown for the genus LINUM, were isolated by HPLC from callus cultures of LINUM CORYMBULOSUM (Linaceae) and identified by spectroscopic methods.

Essential oil of one of the Iranian skullcaps.

The hydro-distilled essential oil from dried aerial parts of one of widespread Iranian skullcaps, Scutellaria pinnatifida A. Hamilt. sap. alpina (Bornm.) Rech. grown in Khorassan province was analyzed by GC and GC/MS. Thirty components were characterized ing 93.8% of the total components detected. The major components of the oil were germacrene-D (39.7%) and beta-caryophyllene (15.0%).