Evolution of the Portulaca oleracea L. aggregate in Egypt on molecular and phenotypic levels revealed by morphology,inter-simple sequence repeat (ISSR) and 18S rDNA gene sequence markers

Molecular markers including inter simple sequence repeats (ISSR) and 18S rDNA gene sequence markers were combined with a detailed morphological analysis to seek characters that discriminate four taxa of Portulaca oleracea s.l. These taxa were identified as the microspecies Portulaca nitida, Portulaca oleracea, Portulaca rausii and Portulaca granulatostellulata. Morphological characters did not provide a clear distinction among the four taxa of P. oleracea s.l. It was found that mixed populations of the taxa occur in several locations and morphological similarities between the microspecies were detected. ISSR analysis indicates that gene flow among populations of different taxa was high and most of the genetic variation (61.9%) occurs within population. These results are inconsistent with the general characteristics of P. oleracea as a self- or 5% cross-pollinated species. A close relationship between P. oleracea, P. rausii and P. granulatostellulata was supported by ISSR and 18S rDNA gene sequence. The ISSR and 18s data support the specific status of P. nitida as a recently evolved taxon. We conclude that P. oleracea exists as a polymorphic species and is not divisible into microspecies based on seed surface as the primary morphological trait, and inter simple sequence repeats (ISSR) and 18S rDNA gene sequence markers. We suggest that the taxa do not merit specific rank as morphological characters and absence of a breeding barrier fail to separate the different populations when they become sympatric.     

Flow cytometric,chromosomal and morphometric analyses challenge current taxonomic concepts in the Portulaca oleracea complex (Portulacaeae,Caryophyllales)

Portulaca oleracea is a noxious annual weed of worldwide distribution in temperate to tropical climates. Its taxonomy has been treated in contradictory ways in the past. Various microspecies have been described, lumped into a single species by other authors. We re‐examined the importance of seed size and ploidy variation, previously applied as the most important taxonomic characters, for systematic classification based on accessions from Europe, Asia, Africa and South America using flow cytometry, chromosome counting and morphometry. Sixteen microspecies and six transitional forms, covering the ploidy and seed character variation, proposed for the complex, were studied from 178 populations. Portulaca grandiflora was included as a reference species from outside the complex. DNA hyper‐pentaploidy or hexaploidy were inferred for the majority of accessions which exhibited the full range of seed size. It is recommended that the only species of lower ploidy (either diploid based on x = 12 or tetraploid based on x = 12) encountered, P. nicaraguensis, should be separated from the P. oleracea complex as it deviates in base chromosome number and monoploid genome size. The frequency distribution of seed size was continuous and unimodal within the wild taxa of the complex and in pairs of taxa defined by testa sculpture. Seed size of DNA hexaploids was slightly negatively correlated with sample/standard fluorescence intensities. Our results conflict with the current microspecies concept. Possible reasons underlying the discrepancy are discussed and strategies for future systematic research are suggested. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 144–156.     

Taxonomic studies of the genusPoa of Japan

With the aim of making clear the boundaries between species in thePoa acroleucahisauchii-nipponica aggregate, chromosomes and morphological features of 746 collections gathered from 125 localities in Japan were examined. For morphological observations, the voucher specimens of 95 collections whose chromosome numbers were reported previously (Tateoka, 1985) were also used. Tetraploids (2n=28) and hexaploids (2n=42), as well as a few pentaploids which were the hybrids between 4× and 6×, were found. By examining morphological features of these collections, two groups were recognized in tetraploids and one in hexaploids. The two tetraploid groups corresponded toPoa acroleuca Steud. andP. hisauchii Honda, and the hexaploid group toP. nipponica Koidz. It was confirmed that the hairiness on the internerve surface of lemma, ligule hairiness and the length ratio of anther/ lemma are the most important features for discriminating between these species; panicle shape, leaf shape and anther length are also helpful for the identification. The ambiguity of the boundaries between species which was hitherto present in the taxonomy of this species aggregate was unrelated to the creation of nature itself but was attributable to the insufficiency of our research work.     

Do polyploids require proportionally less rDNA loci than their corresponding diploids? Examples from Artemisia subgenera Absinthium and Artemisia (Asteraceae,Anthemideae)

Abstract

Fluorescent in situ hybridisation (FISH) of 5S and 18S-5.8S-26S ribosomal DNA was carried out in two species of the genus Artemisia, belonging to the subgenera Artemisia (A. medioxima) and Absinthium (A. lagocephala), each one showing both low and high ploidy levels (2x, 4x and 16x, and 2x and 6x, respectively). Both species have a base chromosome number of x = 9. Linkage of both rDNA genes has been observed confirming previous results. Diploid A. lagocephala (2n = 18) shows three rDNA loci, and the hexaploid six. Also in A. medioxima, the number of rDNA loci does not increase in the proportion given by the ploidy level, and a relative loss is found. In this species, the diploid population shows two rDNA loci, the tetraploid four, and the hexaidecaploid has around 20. The results evidence a relative loss of rDNA loci and heterochromatin, a phenomenon that is more pronounced at higher ploidy levels. Nevertheless, the DAPI banding pattern of A. lagocephala does not follow this trend, as it shows a spectacular increase of heterochromatic bands at the hexaploid level. These results are discussed in the light of possible chromosome restructuring and gene silencing mechanisms that take place during polyploidy, and more especially allopolyploid formation.     

Notes on the reproductive biology ofPortulaca L. (Portulacaceae) in Nigeria

Nigerian species ofPortulaca: P. grandiflora Hooker,P. oleracea L. andP. quadrifida L. combine inbreeding, outbreeding and vegetative propagation in the wild and in the oultivation. Observations have shown that the anthers ofPortulaca species dehisces before the flower bud opens, and are usually ripe at the time the stigmas are receptive. This enhances self-fertilization.     

Phytotoxic effects of Lantana camara, Eucalyptus camaldulensis and Eriocephalus africanus essential oils in weeds of Mediterranean summer crops

The essential oil composition of Lantana camara, Eucalyptus camaldulensis and Eriocephalus africanus was analyzed by means of GC and GC–MS and bioassayed in order to determine their activity against Amaranthus hybridus and Portulaca oleracea. E. camaldulensis essential oil, with spathulenol as the main compound, was the most effective, completely inhibiting germination and seedling growth on both weeds. The essential oil of E. africanus, rich in artemisia ketone, showed activity similar to that of E. camaldulensis on A. hybridus, but it was not so effective against P. oleracea, and L. camara essential oil, with high percentages in sesquiterpene hydrocarbons, also showed higher phytotoxic activity against A. hybridus, inhibiting its germination and seedling length; however, it showed no effect against P. oleracea germination, whereas was effective in significantly reducing its seedling growth at all concentrations assayed. The results suggest the possible use of these essential oils as natural herbicides.     

Integrated analysis for identifying Portulaca oleracea and its sub-species based on chloroplastic and nuclear DNA barcoding

The taxonomy of Portulaca oleracea has been considered as being complex since the aggregate is composed of many subspecies or a group of micro-species based on seed-coat characters, seed size, and chromosome number. In order to enlarge the background of the extent of genetic variability between and within Tunisian P. oleracea accessions, a combined morphological and molecular approach was adapted, in the present survey. The morphological analyses of the spontaneous Tunis population display high intra population variability characterized by two distinct morphotypes corresponding to the botanical forms (wild and cultivated plant). Furthermore, the molecular approach based on sequences data related to chloroplastic and ribosomal DNA, was used to understand this variability. The obtained results highlighted the greater molecular variability of this plant and allowed to segregate between morphotypes and genotypes of Portulaca. Mostly, this work shows the important contribution of DNA barcoding approach in resolving low-level-taxonomy problems to distinguish between natural populations and varieties.     

Weeds ability to phytoremediate cadmium-contaminated soil

An alternative method to other technologies to clean up the soil, air and water pollution by heavy metals is phytoremediation. Therefore, a pot culture experiment was conducted at the College of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran, in 2014 to determine the potential absorption of cadmium by Portulaca oleracea (Common purslane), Solanum nigrum (Black nightshade), Abutilon theophrasti (Velvetleaf) and Taraxacum officinale (Dandelion). The type of experiment was completely randomized design with factorial arrangement and four replications. The soil in pot was treated with different rates of CdCl₂.H₂O (0 (control), 10, 20, 40, 60, and 80 mg Cd/kg soil) and the plants were sown. With increasing concentration levels, fresh weight and dry weight of shoots and roots of all plant species were reduced. The reduction severity was ranked according the following order, P. oleracea > A. theophrasti > S. nigrum > T. officinale. Bioconcentration factor (BCF), Translocation factor (TF) and Translocation efficiency (TE%) was ranked according the following order, T. officinale > S. nigrum > A. theophrasti > P. oleracea. The results of this study revealed that T. officinale and S. nigrum are effective species to phytoremediate Cd-contaminated soil.     

Evaluation of Atriplex Halimus,Medicago Lupulina and Portulaca Oleracea For Phytoremediation of Ni,Pb, and Zn

Suitable plant species are able to accumulate heavy metals and to produce biomass useful for non-food purposes. In this study, three endemic Mediterranean plant species, Atriplex halimus, Portulaca oleracea and Medicago lupulina were grown hydroponically to assess their potential use in phytoremediation and biomass production. The experiment was carried out in a growth chamber using half strength Hoagland's solutions separately spiked with 5 concentrations of Pb and Zn (5, 10, 25, 50, and 100 mg L^?1), and 3 concentrations of Ni (1, 2 and 5 mg L^?1). Shoot and root biomass were determined and analyzed for their metals contents. A. halimus and M. lupulina gave high shoot biomass with relatively low metal translocation to the above ground parts. Metals uptake was a function of both metals and plant species. It is worth noting that M. lupulina was the only tested plant able to grow in treatment Pb50 and to accumulate significant amount of metal in roots. Plant metal uptake efficiency ranked as follows: A. halimus > M. lupulina > P. oleracea . Due to its high biomass production and the relatively high roots metal contents, A. halimus and M. lupulina could be successfully used in phytoremediation, and in phytostabilization, in particular.     

Comparison of CAM expression,photochemistry and antioxidant responses in Sedum album and Portulaca oleracea under combined stress

Previous studies of crassulacean acid metabolism (CAM) pathway during stress have been directed at individual drought and salinity stress, here, we studied the effects of a combination of drought and salt on CAM expression, chlorophyll fluorescence and antioxidant parameters in the C₃-CAM facultative Sedum album and C₄-CAM facultative Portulaca oleracea plants. While salinity alone was not able to induce functional CAM expression in P. oleracea leaves, we showed that salinity induced low level of nocturnal acid accumulation in S. album species. After 20 d of exposure to the combination of simultaneous salt and drought stress, P. oleracea plants exhibited more resistance to photoinhibition as compared to S. album plants. The decrease of maximum quantum yield (F_v/F_m) in S. album leaves under combined stress was in parallel with the largest suppression of CAM expression of >50%, probably displaying the withdrawal of functional CAM back to C₃ pathway. However, under drought treatment alone, S. album plants exhibited higher photosynthetic flexibility, which was associated with the up-regulation of antioxidant enzymes activities and maintenance of glutathione (GSH) pool, and consequently higher photochemical functioning. The levels of nitric oxide (NO) correlated well with CAM expression, which was observed only in S. album, suggesting that NO acts in a different way in C₃ and C₄ species during CAM induction. Additionally, in both species, over the course of CAM induction, the changes in CAM expression parameters exhibited a similar pattern to that of antioxidant capacity and photochemical functioning parameters.     

Life table parameters of large white butterfly Pieris brassicae (Lepidoptera: Pieridae) on different cabbage varieties

Effect of four different cole crops (Brassica oleracea var. botrytis, Brassica oleracea var. capitata, Brassica oleracea var. italica and Brassica oleracea var. viridis) on biological parameters of the large white butterfly Pieris brassicae was evaluated at temperature 26 ± 1 °C, 60 ± 5% R. H. and a photoperiod of 16: 8 (L:D) h. The shortest larval and pupal period stages were recorded on B. oleracea var. botrytis (22.18 ± 0.20 days) and (13.32 ± 0.17 days), respectively. The life span was longest on B. oleracea var. viridis (60.43 ± 2.34 days) and shortest on B. oleracea var. botrytis (50.19 ± 0.51 days). The highest percentage of larval and pupal mortality was observed on B. oleracea var. viridis (74%), and (53%), respectively. We found that P. brassicae prefers B. oleracea var. botrytis and B. oleracea var. capitata among cole crops; it is due to the lowest percentage of larval and pupal mortality and the highest rate of life table parameters, including survival rate (lx) and life expectancy (ex), which makes them to be susceptible varieties to this pest. Contrary, a longer developmental time on B. oleracea var. viridis may be attributed to poor nutritional status and reduced survival of the cohort, resulting in high rates of mortality, which was partial resistance to pest. Knowledge of the biology and life table parameters of P. brassicae on different cole crops could be effective in detecting and monitoring the pest infestation, variety selection and crop breeding.     

Susceptibility of Several Common Subtropical Weeds to Meloidogyne arenaria,M. incognita,and M. javanica

Experiments were conducted in the greenhouse to assess root galling and egg production of three root-knot nematode species, Meloidogyne arenaria, M. incognita, and M. javanica, on several weeds common to Florida agricultural land. Weeds evaluated were Amaranthus retroflexus (redroot pigweed), Cyperus esculentus (yellow nutsedge), Eleusine indica (goosegrass), Portulaca oleracea (common purslane), and Solanum americanum (American black nightshade). Additionally, although it is recommended as a cover crop in southern regions of the U.S., Aeschynomene americana (American jointvetch) was evaluated as a weed following the detection of root galling in a heavy volunteer infestation of an experimental field in southeastern Florida. Weeds were propagated from seed and inoculated with 1000 nematode eggs when plants reached the two true-leaf stage. Tomato (Solanum lycopersicum ‘Rutgers’) was included as a positive control. Aeschynomene americana and P. oleracea roots supported the highest number of juveniles (J₂) and had the highest number of eggs/g of root for all three species of Meloidogyne tested. However, though P. oleracea supported very high root levels of the three nematode species tested, its fleshy roots did not exhibit severe gall symptoms. Low levels of apparent galling, combined with high egg production, increase the potential for P. oleracea to support populations of these three species of root-knot nematodes to a degree that may not be appropriately recognized. This research quantifies the impact of P. oleracea as a host for M. arenaria, M. incognita, and M. javanica compared to several other important weeds commonly found in Florida agricultural production, and the potential for A. americana to serve as an important weed host of the three species of root-knot nematode tested in southern regions of Florida.     

Bellevalia juliana (Asparagaceae), a new hexaploid species from E Kriti (Greece)

A new species, Bellevalia juliana Bareka, Turland & Kamari (Asparagaceae) is described from E Kriti (Greece) and compared with other related taxa from Greece and the Mediterranean area. It is a member of B. sect. Bellevalia and a karyological study revealed a hexaploid karyotype with 2n = 6x = 24 chromosomes, illustrated here. The conservation status of the new species is assessed as Vulnerable (VU), according to IUCN criteria.     

Brassica taxonomy based on nuclear restriction fragment length polymorphisms (RFLPs)

Summary RFLPs were used to study genome evolution and phylogeny in Brassica and related genera. Thirtyeight accessions, including 10 accessions of B. rapa (syn. campestris), 9 cultivated types of B. oleracea, 13 nine-chromosome wild brassicas related to B. oleracea, and 6 other species in Brassica and allied genera, were examined with more then 30 random genomic DNA probes, which identified RFLPs mapping to nine different linkage groups of the B. rapa genome. Based on the RFLP data, phylogenetic trees were constructed using the PAUP microcomputer program. Within B. rapa, accessions of pak choi, narinosa, and Chinese cabbage from East Asia constituted a group distinct from turnip and wild European populations, consistent with the hypothesis that B. rapa had two centers of domestication. A wild B. rapa accession from India was positioned in the tree between European types and East Asian types, suggesting an evolutionary pathway from Europe to India, then to South China. Cultivated B. oleracea morphotypes showed monophyletic origin with wild B. oleracea or B. alboglabra as possible ancestors. Various kales constitute a highly diverse group, and represent the primitive morphotypes of cultivated B. oleracea from which cabbage, broccoli, cauliflower, etc. probably have evolved. Cauliflower was found to be closely related to broccoli, whereas cabbage was closely related to leafy kales. A great diversity existed among the 13 collections of nine-chromosome wild brassicas related to B. oleracea, representing various taxonomic states from subspecies to species. Results from these studies suggested that two basic evolutionary pathways exist for the diploid species examined. One pathway gave rise to B. fruticulosa, B. nigra, and Sinapis arvensis, with B. adpressa or a close relative as the initial ancestor. Another pathway gave rise to B. oleracea and B. rapa, with Diplotaxis erucoides or a close relative as the initial ancestor. Raphanus sativus and Eruca sativus represented intermediate types between the two lineages, and might have been derived from introgression or hybridization between species belonging to different lineages. Molecular evidence for an ascending order of chromosome numbers in the evolution of Brassica and allied genera was obtained on the basis of RFLP data and phylogenetic analysis.     

Oxalate depletion from leafy vegetables using alginate entrapped banana oxalate oxidase

Summary The feasibility of depleting the high oxalate content of spinach (Spinacia oleracea), amaranthus (Amaranthus gangeticus), and paruppu keerai (Portulaca oleracea) using alginate entrapped banana oxalate oxidase is demonstrated for the first time. The relevance of this work to low-oxalate diet preparation for hyperoxaluric patients is discussed.     

Re-consideration of Peronospora farinosa infecting Spinacia oleracea as distinct species, Peronospora effusa

Downy mildew is probably the most widespread and potentially destructive global disease of spinach (Spinacia oleracea). The causal agent of downy mildew disease on various plants of Chenopodiaceae, including spinach, is regarded as a single species, Peronospora farinosa. In the present study, the ITS rDNA sequence and morphological data demonstrated that P. farinosa from S. oleracea is distinct from downy mildew of other chenopodiaceous hosts. Fifty-eight spinach specimens were collected or loaned from 17 countries of Asia, Europe, Oceania, North and South America, which all formed a distinct monophyletic group. No intercontinental genetic variation of the ITS rDNA within Peronospora accessions causing spinach downy mildew disease was found. Phylogenetic trees supported recognition of Peronospora from spinach as a separate species. Microscopic examination also revealed morphological differences between Peronospora specimens from Spinacia and P. farinosa s. lat. specimens from Atriplex, Bassia, Beta, and Chenopodium. Consequently, the name Peronospora effusa should be reinstated for the downy mildew fungus found on spinach. Here, a specimen of the original collections of Peronospora effusa is designated as lectotype.     

马齿苋内生菌橘青霉和波兰青霉中抗青枯菌的活性物质

为了挖掘农作物病害生物防治新资源,以药用植物马齿苋(Portulaca oleracea)为材料,通过培养基种植法分离和纯化其根、茎、叶中的内生菌,以青枯菌(Ralstonia solanacearum)的抑菌试验评价其活性,采用菌落形态观察和ITS序列分析鉴定菌种。结果表明,从马齿苋筛选出2种具有抑制青枯菌的内生菌橘青霉(Penicillium citrinum)和波兰青霉(P. polonicum),采用液相与四极杆飞行时间串联质谱(UPLC-QTOF-MS)鉴定2种内生菌的主要活性物质为橘霉素,其对青枯菌的抑制效果比链霉素更好。因此,这为植物青枯病的生物防治提供科学依据。     

Nonea dumanii sp. nov. (Boraginaceae) from the Taurus mountains (south Turkey)

The new species Nonea dumanii, endemic to the mountains of the western Taurus in south Turkey (C4 Antalya), is described an illustrated based on original collections by the authors. Karyological observations and analysis of ITS1 DNA sequences showed that the species is probably hexaploid with 2n = 6x = 60 and has phylogenetic affinity to the diploid N. monticola from the Paphlagonian mountains as well as to the tetraploid N. anchusoides from northwest Iran and southeast Turkey. From these allopatric species it is morphologically distinct in characters of the indumentum, flower and fruit. Polyploidy is supported as a major driving force for speciation in Nonea, especially in the group of Anatolian mountain species with primary base number x = 10.     

Relationship between ploidy level and genome size in strawberries

This study investigated the pattern of variation in nuclear DNA content at different ploidy levels in Fragaria (Strawberry, Rosaceae) using flow cytometry based on mean fluorescent intensity (MFI) reflected by propidium-iodide-stained nuclei. On average, MFI values were 237 for diploids F. vesca, F. viridis, and F. nubicola, 416.5 for tetraploid F. orientalis, 621.5 for hexaploid F. moschata, and 798 for octoploids F. × ananassa, F. virginiana, and F. chiloensis. Within diploids MFI ranged from 225.9 in F. vesca ssp. vesca to 255.4 in F. nubicola, and within octoploids varied from 766 in F. × ananassa to 808 in F. virginiana. The nuclear DNA variation was significant among diploid species (N = 21, P < 0.008), but not across octoploid species (N = 17, P>0.386). MFI values were also variable among different genotypes of a given species though not significant. The values of mean basic genome DNA (MFI divided by ploidy level) were 118.5, 104, 103.5, and 99.8, respectively, for diploids, tetraploid, hexaploid, and octoploid species. This indicates that relative genomic size decreases by increasing ploidy level, and that there is no direct proportional relationship between DNA content and ploidy levels in Fragaria, supporting the idea of genome downsizing during polyploidization in plants.     

Genome identification of the Triticum crassum complex (Poaceae) with the restriction patterns of repeated nucleotide sequences

Species of the Triticum crassum complex, tetraploid and hexaploid T. crassum, hexaploid T. juvenale, and hexaploid T. syriacum, share a similar morphology. Variation in the restriction profiles of nuclear repeated nucleotide sequences is employed in identification of genomes of these species. The data show that hexaploid T. crassum originated from hybridization of the tetraploid cytotype of T. crassum with T. tauschii. Triticum juvenale and T. syriacum originated from hybridization of tetraploid T. crassum with T. umbellulatum and T. searsii, respectively. Tetraploid T. crassum appears to be an ancient allotetraploid that originated from hybridization of primitive T. tauschii with an ancient species in the evolutionary lineage leading to the section Sitopsis of the genus Triticum.