Assessment of antioxidant and antimicrobial activities of Crambe spp. during vegetation.

BACKGROUND: The search for new plant raw material as a potential source of antioxidants is still ongoing. This study aimed to evaluate the antioxidant and antimicrobial capacity of the plant raw material of Crambe spp. during vegetation. METHODS: The free radical scavenging activity and molybdenum reducing power of the extracts were used to determine antioxidant activity. The quantification of polyphenol compounds was conducted with Folin-Ciocalteu reagent. Flavonoids and phenolic acids were also determined. The disc diffusion method was used to determine antimicrobial activity. RESULTS: It was determined that the free radical scavenging activity, assessed using the DPPH-method, was 4.38-8.20 mg TE/g DW, the molybdenum reducing power of the extracts was 40.07-129.12 mg TE/g DW, total polyphenol content was 20.24-70.88 mg GAE/g DW, total flavonoid content was 5.73-29.92 mg QE/g DW, and phenolic acid content was 3.00-10.63 mg CAE/g DW. Antimicrobial activity depended on the stage of growth and the part of the plant used. CONCLUSIONS: Crambe spp. possess the antioxidant and antimicrobial potential to mean that they could be used in pharmaceutical studies and the food industry.

Complete chloroplast genome features and phylogenetic analysis of Eruca sativa (Brassicaceae).

Eruca sativa Mill. (Brassicaceae) is an important edible vegetable and a potential medicinal plant due to the antibacterial activity of its seed oil. Here, the complete chloroplast (cp) genome of E. sativa was de novo assembled with a combination of long PacBio reads and short Illumina reads. The E. sativa cp genome had a quadripartite structure that was 153,522 bp in size, consisting of one large single-copy region of 83,320 bp and one small single-copy region of 17,786 bp which were separated by two inverted repeat (IRa and IRb) regions of 26,208 bp. This complete cp genome harbored 113 unique genes: 79 protein-coding genes, 30 tRNA genes, and four rRNA genes. Forty-nine long repetitive sequences and 69 simple sequence repeats were identified in the E. sativa cp genome. A codon usage analysis of the E. sativa cp genome showed a bias toward codons ending in A/T. The E. sativa cp genome was similar in size, gene composition, and linearity of the structural region when compared with other Brassicaceae cp genomes. Moreover, the analysis of the synonymous (Ks) and non-synonymous (Ka) substitution rates demonstrated that protein-coding genes generally underwent purifying selection pressure, expect ycf1, ycf2, and rps12. A phylogenetic analysis determined that E. sativa is evolutionarily close to important Brassica species, indicating that it may be possible to transfer favorable E. sativa alleles into other Brassica species. Our results will be helpful to advance genetic improvement and breeding of E. sativa, and will provide valuable information for utilizing E. sativa as an important resource to improve other Brassica species.

Microscopic investigation of pollen morphology of Brassicaceae from Central Punjab-Pakistan.

The intention of the current study is to provide an account on the palynological features of Brassicaceae from Central Punjab-Pakistan as a basis for future studies. Different morpho-palynological characteristics both qualitative and quantitative were analyzed during this research which includes shape of pollen, diameter of pollen, P/E ratio, exine sculpturing, thickness of exine, type of pollen, shape and size of lumens, and thickness of murus. Taxonomic keys were also constructed based on pollen morphological characters for correct identification of species. This study aims to provide detailed information of pollen diversity and their exine structure based on both qualitative and quantitative characters by using Light microscopy and Scanning electron microscopy. Shape of pollen is mostly prolate, but some species also have sub-prolate to spheroidal prolate types. Exine ornamentation in most species was reticulate, whereas micro reticulate (one species) and coarsely reticulate (one species) exine also observed in some pollen. All the pollen mentioned in this study have tricolpate apertures. Variation found in thickness of exine and other characters proved to be helpful at generic and specific level. The results reinforced the significance of pollen morphological features of family Brassicaceae and aid for valuable taxonomic tool in plant systematics.

Taxonomic Importance of Pollen Morphology for Some Species of Brassicaceae.

BACKGROUND AND OBJECTIVE: Pollen morphology is one of the significant tools in solving some taxonomic problems on the family, generic or specific level and has become part of the multidisciplinary and collaborative approach in plant systematic and evolution. Therefore, this study aimed to investigate and describe the pollen morphology of 10 species belongs to 9 genera and five tribes of Brassicaceae from eastern region of Saudi Arabia by using light and scanning electron microscope. MATERIALS AND METHODS: To study the pollen morphology for ten species representing 9 genera and 5 tribes of Brassicaceae the Light Microscope (LM) and Scanning Electron Microscope (SEM). For the SEM the pollen was placed directly on brass stubs without treatment and mounted onto a metallic stub with a double-sided adhesive tape. Gold coating of few nanometers was applied using sputter coating machine (Quorum, Q150R ES, UK) to avoid charging and capture high quality images. Two statistical program; PRIMER 6, version 6.1.6 and SPSS version 16are used to fine the relationships among the studied species. RESULTS: The shape of pollen grains recorded three types; prolate, subprolate and prolate-spheroidal, the main types were prolate. The apertures were tricolpate in all studied species. Exine ornamentation of studied species recorded two types; reticulate and coarsely reticulate. The results of numerical analysis showed that species were grouped into two major clusters and each cluster divided into two groups. CONCLUSION: The results show that the morphology of pollen grains cannot be useful for taxonomical classification of the tribes but can be useful for differentiate between species belong to the same genera.

Phytochemical profiling of five medicinally active constituents across 14 Eutrema species.

Wasabi or Japanese horseradish (Eutrema japonicum) is both a traditional condiment and a medicinally important plant with diverse uses. Its medicinally active constituents appear to include five isothiocyanates, but their spatial variations in naturally occurring congeners are unknown. Thus, in this study we measured concentrations of these five active constituents in 20 populations of 14 species of Eutrema and one related species, Yinshania sinuata. Three to five of these constituents were detected in each of the examined species, at concentrations that varied greatly between sampled species and populations of the same species. However, two species, Eutrema tenue and Eutrema deltoideum, had higher total concentrations of the five isothiocyanates and substantially higher concentrations of one or two, than the widely cultivated E. japonicum. Thus, both of these species could be important wild resources for artificial cultivation, in addition to the currently widely cultivated E. japonicum.

Phytochemical and Biological Investigation of Two Diplotaxis Species Growing in Tunisia: D. virgata & D. erucoides.

A phytochemical investigation of Diplotaxis virgata D.C. and D. erucoides (L.) D.C. (Brassicaceae) offered to the isolation of two new flavonoids isorhamnetin-3-O-α-l-glucopyranoside (1) and rhamnetin-3,3'-di-O-ß-d-glucopyranoside (2), respectively. Their structures have been elucidated from the extended spectroscopic methods, including 1D- and 2D-NMR, UV and mass spectrometry analysis and by comparison with literature data. The fatty acid composition of the hexane extracts of the two species was also investigated by using GC-MS. The antioxidant activity of ethanol, ethyl acetate, n-butanol extracts and the isolated compounds from the two species was evaluated using DPPH and ABTS⁺ scavenging assays. All the tested samples showed an efficient radical scavenging ability, with IC50 values ranging from 16-40 µg/mL for the DPPH and from 17-44 µg/mL for the ABTS⁺ assays. In addition, the antibacterial activity of the prepared extracts and compounds 1 and 2, determined by well diffusion agar method against two Gram positive and five Gram negative bacteria, was evaluated and the results showed significant effects against all strains used.

Production and genetic characterization of interspecific hybrids among Crambe abyssinica, Crambe hispanica and Crambe kralikii.

In this paper, interspecific crosses among Crambe abyssinica, Crambe hispanica, and Crambe kralikii were reported. In the C. hispanica x C. abyssinica (H x A) cross, 118 F1 hybrids were produced without embryo rescue, while 5 F1 hybrids were obtained with embryo rescue, when C. hispanica was used as the female parent. In the reciprocal cross (A x H), 232 hybrids were obtained without embryo rescue. From more than 1000 C. kralikii flowers pollinated with pollen grains of C. abyssinica (K x A), only 2 F1 hybrids were obtained with embryo rescue, whereas the reciprocal cross produced no hybrids, even with embryo rescue. The hybrids were confirmed at the morphological, cytological, and molecular levels. In the combinations of A x H and H x A, many BC1 hybrids were obtained without embryo rescue. In contrast, in the K x A cross, only 7 BC1 plants were obtained with embryo rescue, while no seed set was achieved under self-pollination or in backcrosses without embryo rescue. In the H x A F1 hybrids, the pollen stainability was 65.4-86.0%, with an average of 76.9%. In comparison, the pollen viability of hybrids in the reciprocal cross (A x H) ranged from 66.2 to 81.1%, with an average of 75.4%. Fertile pollen grains were not found in the K x A F1 hybrids. All F1 hybrids of the 3 crosses (H x A, A x H, and K x A) had the expected 2n = 75 chromosomes. AFLP analyses indicated that all F1 hybrids and their progenies had typical bands of the parents. These hybrids and progenies are anticipated to be valuable for future C. abyssinica improvement in breeding programs.

Characterization of selenium and sulfur accumulation across the genus Stanleya (Brassicaceae): A field survey and common-garden experiment.

UNLABELLED: • PREMISE OF STUDY: Selenium (Se) hyperaccumulation, the capacity to concentrate the toxic element Se above 1000 mg·kg(-1)·dry mass, is found in relatively few taxa native to seleniferous soils. While Se hyperaccumulation has been shown to likely be an adaptation that protects plants from herbivory, its evolutionary history remains unstudied. Stanleya (Brassicaceae) is a small genus comprising seven species endemic to the western United States. Stanleya pinnata is a hyperaccumulator of selenium (Se). In this study we investigated to what extent other Stanleya taxa accumulate Se both in the field and a greenhouse setting on seleniferous soil.• METHODS: We collected multiple populations of six of the seven species and all four varieties of S. pinnata We tested leaves, fruit, and soil for in situ Se and sulfur (S) concentrations. The seeds collected in the field were used for a common garden study in a greenhouse.• KEY RESULTS: We found that S. pinnata var. pinnata is the only hyperaccumulator of Se. Within S. pinnata var. pinnata, we found a geographic pattern related to Se hyperaccumulation where the highest accumulating populations are found on the eastern side of the continental divide. We also found differences in genome size within the S. pinnata species complex.• CONCLUSIONS: The S. pinnata species complex has a range of physiological properties making it an attractive system to study the evolution of Se hyperaccumulation. Beyond the basic scientific value of understanding the evolution of this fascinating trait, we can potentially use S. pinnata or its genes for environmental cleanup and/or nutrient-enhanced dietary material.

Cytological studies of Brassicaceae burn. (Cruciferae juss.) from Western Himalayas.

Cytological studies have been carried out on 12 species of Brassicaceae Burn. on population basis from different geographical areas of Kashmir and Himachal Pradesh in the Western Himalayas. Variable chromosome reports for Barbaraea intermedia (n = 16), Cardamine loxostemonoides (n = 8), Nasturtium officinale (n = 8), Sisymbrium orientale (n = 14) on world-wide basis have been added to the previous reports of these species. The chromosome numbers in seven species as Barbaraea intermedia (n = 8), B. vulgaris (n = 8), Capsella bursa-pastoris (n = 8), Descuriania sophia (n = 10), Rorippa islandica (n = 8), Sisymbrium strictum (n = 7) and Thlaspi alpestre (n = 7) have been worked out for the first time from India. The meiotic course in the populations of seven species such as Barbaraea intermedia, Capsella bursa-pastoris, Coronopus didymus, Descuriania sophia, Nasturtium officinale, Sisymbrium orientale and S. strictum varies from normal to abnormal while all the populations of two species Barbaraea vulgaris and Sisymbrium irio show abnormal meiotic course. Meiotic abnormalities are in the form of cytomixis, chromosomal stickiness, unoriented bivalents, inter-bivalent connections, formation of laggards and bridges, all resulting into abnormal microsporogenesis. Heterogenous sized fertile pollen grains and reduced reproductive potentialities have invariably been observed in all the meiotically abnormal populations. However, the meiotic course in all the populations of Cardamine loxostemonoides, Rorippa islandica and Thalspi alpestre is found to be normal with high pollen fertility.

Biogeographic distribution of polyploidy and B chromosomes in the apomictic Boechera holboellii complex.

The Boechera holboellii complex comprises B. holboellii and B. drummondii, both of which can reproduce through sex or apomixis. Sexuality is associated with diploid individuals, whereas apomictic individuals are diploid or triploid and may additionally have B chromosomes. Using flow cytometry and karyotype analysis, we have shown that B chromosomes (a) occur in both diploid and triploid apomictic B. holboellii, (b) may occur in triploid B. drummondii, and (c) are dispensable for the plant. Both diploid and triploid karyotypes are found in multiple chloroplast haplotypes of both species, suggesting that triploid forms have originated multiple times during the evolution of this complex. B chromosome carriers are found in geographically and genetically distinct populations, but it is unknown whether the extra chromosomes are shared by common descent (single origin) or have originated via introgressive hybridization and repeated transitions from diploidy to triploidy. Diploid plants containing the Bs reproduce apomictically, suggesting that the supernumerary elements are associated with apomixis. Finally, our analyses of pollen size and viability suggest that irregular chromosome segregation in some triploid lineages may lead to the generation of diploid individuals which carry the B chromosomes.

Metabolism and detoxification of phytoalexins and analogs by phytopathogenic fungi.

To date, the many examples reporting that fungal pathogens can efficiently detoxify phytoalexins provide strong evidence that the pathogenicity and/or virulence of some fungi is linked to their ability to detoxify their hosts' phytoalexins. The pathways used by plant pathogenic fungi to metabolize and detoxify phytoalexins are reviewed. Prospects for application of recent findings are discussed.

Comparisons of pollen coat genes across Brassicaceae species reveal rapid evolution by repeat expansion and diversification.

Reproductive genes and traits evolve rapidly in many organisms, including mollusks, algae, and primates. Previously we demonstrated that a family of glycine-rich pollen surface proteins (GRPs) from Arabidopsis thaliana and Brassica oleracea had diverged substantially, making identification of homologous genes impossible despite a separation of only 20 million years. Here we address the molecular genetic mechanisms behind these changes, sequencing the eight members of the GRP cluster, along with 11 neighboring genes in four related species, Arabidopsis arenosa, Olimarabidopsis pumila, Capsella rubella, and Sisymbrium irio. We found that GRP genes change more rapidly than their neighbors; they are more repetitive and have undergone substantially more insertion/deletion events while preserving repeat amino acid composition. Genes flanking the GRP cluster had an average K(a)/K(s) approximately 0.2, indicating strong purifying selection. This ratio rose to approximately 0.5 in the first GRP exon, indicating relaxed selective constraints. The repetitive nature of the second GRP exon makes alignment difficult; even so, K(a)/K(s) within the Arabidopsis genus demonstrated an increase that correlated with exon length. We conclude that rapid GRP evolution is primarily due to duplication, deletion, and divergence of repetitive sequences. GRPs may mediate pollen recognition and hydration by female cells, and divergence of these genes could correlate with or even promote speciation. We tested cross-species interactions, showing that the ability of A. arenosa stigmas to hydrate pollen correlated with GRP divergence and identifying A. arenosa as a model for future studies of pollen recognition.

Rapid evolution of a pollen-specific oleosin-like gene family from Arabidopsis thaliana and closely related species.

It has been shown in a variety of species that genes expressed in reproductive tissues evolve rapidly, which often appears to be the result of positive Darwinian selection. We investigated the evolution of a family of seven pollen-specific oleosin-like proteins (or oleopollenins) in Arabidopsis thaliana and two closely related species. More than 30 kb of a genomic region that harbors the complete, tandemly repeated oleopollenin cluster were sequenced from Arabidopsis lyrata ssp. lyrata, and Boechera drummondii. A phylogenetic analysis of the complete gene cluster from these three species and from Brassica oleracea confirmed its rapid evolution resulting from gene duplication and gene loss events, numerous amino acid substitutions, and insertions/deletions in the coding sequence. Independent duplications were inferred in the lineages leading to Arabidopsis and to Brassica, and gene loss was inferred in the lineage leading to B. drummondii. Comparisons of the ratio of nonsynonymous (d(N)) and synonymous (d(S)) divergence revealed that the oleopollenins are among the most rapidly evolving proteins currently known from Arabidopsis and that they may evolve under positive Darwinian selection. Reverse transcriptase polymerase chain reaction analysis demonstrated the expression of oleopollenins in flowers of the outcrossing A. lyrata, the selfing B. drummondii, and the apomictic Boechera holboellii, suggesting that oleopollenins play an important role in species with different breeding systems. These results are consistent with a putative function in species recognition, but further analyses of protein function and sequence variation in species with different breeding systems are necessary to reveal the underlying causes for the rapid evolution of oleopollenins.

[Quality evaluation of indigowoad root and leaf by chemical pattern recognition].

27 samples of indigowoad root and 5 samples of indigowoad leaf from different species and in different habital were collected. The chemical constituents were analyzed by HPLC, and ten original chemical features as a whole obtained. Meanwhile, extrinsic bacteriostasis experiments were carried out for each of the 32 samples. Canonical correlation analysis was applied to the intercorrelation between the chemical and the pharmacologic data, and five chemical features were found to be the effective constituents. In the end, the 32 samples were divided into five classes by the fuzzy clustering technique ISODATA(interachive self-organizing data analysis technique A).