Primotrapa gen. nov., an extinct transitional genus bridging the evolutionary gap between Lythraceae and Trapoideae, from the early Miocene of North China.

BACKGROUND: Although Trapa is a well-defined genus of distinctive freshwater plants with accumulations of extensive morphological and embryological autapomorphies, its phylogenetic relationships have long been unclear. Formerly placed in the monotypic family Trapaceae, Trapa is now recognized as sister to Sonneratia within Lythraceae s.l., although both genera lack morphological synapomorphies. Thus, a split between the two taxa must have occurred in deep evolutionary time, which raises the possibility of finding transitional forms in the fossil record. RESULTS: Here we describe a new genus and species, Primotrapa weichangensis Y. Li et C.-S. Li (Lythraceae s.l.: Trapoideae), based on three-dimensionally preserved floral cups, fruits, and seeds from the early Miocene of Weichang County, Hebei Province, China. Primotrapa is characterized by a shallow, saucer-shaped floral cup, four distally barbellate sepals, four intersepal appendages alternating with the sepals at the rim of cup, a superior to basally inferior ovary, a fusiform or ovoid, one-seeded fruit with a ribbed surface, and a long persistent peduncle. Two fossil species of Hemitrapa are proposed as new combinations of Primotrapa, namely P. alpina (T. Su et Z.-K. Zhou) Y. Li et C.-S. Li comb. nov. and P. pomelii (Boulay) Y. Li et C.-S. Li comb. nov. Our phylogenetic analysis based on fifteen flower and fruit characters supports the placement of Primotrapa, Hemitrapa and Trapa in a monophyletic clade, which comprise subfamily Trapoideae. The phylogenetic analysis places Primotrapa at the base of Trapoideae. CONCLUSIONS: In view of its superior ovary, which is a plesiomorphic character of Lythraceae s.l., the newly recognized genus Primotrapa and its three species likely represent transitional forms that bridge the evolutionary gap between the basal taxa of Lythraceae s.l., i.e. Lythrum, and the highly derived taxon Trapa.

The Complete Chloroplast Genomes of Punica granatum and a Comparison with Other Species in Lythraceae.

Pomegranates (Punica granatum L.) are one of the most popular fruit trees cultivated in arid and semi-arid tropics and subtropics. In this study, we determined and characterized three complete chloroplast (cp) genomes of P. granatum cultivars with different phenotypes using the genome skimming approach. The complete cp genomes of three pomegranate cultivars displayed the typical quadripartite structure of angiosperms, and their length ranged from 156,638 to 156,639 bp. They encoded 113 unique genes and 17 are duplicated in the inverted regions. We analyzed the sequence diversity of pomegranate cp genomes coupled with two previous reports. The results showed that the sequence diversity is extremely low and no informative sites were detected, which suggests that cp genome sequences may be not be suitable for investigating the genetic diversity of pomegranate genotypes. Further, we analyzed the codon usage pattern and identified the potential RNA editing sites. A comparative cp genome analysis with other species within Lythraceae revealed that the gene content and organization are highly conserved. Based on a site-specific model, 11 genes with positively selected sites were detected, and most of them were photosynthesis-related genes and genetic system-related genes. Together with previously released cp genomes of the order Myrtales, we determined the taxonomic position of P. granatum based on the complete chloroplast genomes. Phylogenetic analysis suggested that P. granatum form a single clade with other species from Lythraceae with a high support value. The complete cp genomes provides valuable information for understanding the phylogenetic position of P. gramatum in the order Myrtales.

Targeted metabolite profiling to gain chemometric insight into Indian pomegranate cultivars and elite germplasm.

BACKGROUND: Pomegranate fruit is an excellent source of bioactive polyphenolics, known to contribute significantly to human health. India is the largest producer of pomegranate in the world and produces the finest quality fruit with highly desirable consumer traits such as soft seeds, low acidity, and attractive fruit and aril color. Knowledge of the extent of variation in key metabolites (sugars, organic acids, phenolics, and anthocyanins) is key to selecting superior genotypes for germplasm improvement. Relevant information with respect to Indian genotypes is scarce. The present study therefore aims to evaluate quantitatively important metabolites in some cultivars and elite germplasm of pomegranate in India. RESULTS: Identification and quantification of primary and secondary metabolites such as sugars, organic acids, vitamin C, polyphenolics, and anthocyanins were conducted using a liquid chromatography - mass spectrometry (LC-MS) platform. Fructose and citric acid were the predominant sugar and organic acid, respectively. Wild genotypes had significantly higher concentrations of organic acids, antioxidant activity, and phenolics, namely punicalagin, ellagic acid, sinapic, and ferulic acid. CONCLUSION: Cyanidin and delphinidin derivatives of anthocyanins were more abundant in red aril commercial genotypes. Results suggest that wild-sour accessions represent a rich source of polyphenolics that can be utilized in future breeding programs to breed healthier varieties, food supplements, and pharmaceutical products. © 2019 Society of Chemical Industry.

Quantification of punicalagins in commercial preparations and pomegranate cultivars, by liquid chromatography-mass spectrometry.

BACKGROUND: Pomegranate (Punica granatum L.) - a delicious fruit once used in Ayurvedic medicine - is now largely known for the antioxidant properties of its juice, which has also been considered to have health benefits against diseases such as cancer and cardiovascular diseases. These beneficial effects are associated with the fruit's high content of polyphenolic compounds. High demand and lower production levels drive pomegranate prices up, which leads to the possibility of pomegranate products being adulterated, diluted or substituted. To ensure the presence of pomegranate in various preparations labeled as containing pomegranate, a simple method was developed to screen and quantify the specific punicalagins by mass spectrometry. RESULTS: The present method was used to analyze several pure and mixed beverages from the US market, and also to quantify punicalagins in the juice of 14 pomegranate cultivars. Punicalagins were detected in all cultivars, with higher concentrations in whole fruit juices compared with aril juices. Amongst the 20 commercial beverages, punicalagins were not detected in four preparations. CONCLUSION: The liquid chromatographic-mass spectrometric method presented herein enables an easy and rapid quantification of the specific punicalagins. The latter was detected in all cultivar samples, thus supporting that punicalagin is a suitable marker of these 14 pomegranate cultivars in commercial juices. Absence of the specific marker in four commercial preparations shows the necessity of having simple and rapid methods to evaluate the presence of pomegranate in preparations. © 2019 Society of Chemical Industry.

Diversity among Pomegranate Varieties in Chilling Tolerance and Transcriptome Responses to Cold Storage.

We found great variability in chilling tolerance among 84 pomegranate varieties from the Newe Ya'ar collection; among them, 'Ganesh' was chilling-sensitive, whereas 'Wonderful' was relatively chilling-tolerant. To evaluate the different molecular responses of these varieties to cold storage, we analyzed the transcriptomic changes in the inner membrane tissues of 'Ganesh' and 'Wonderful' fruit after 2 weeks of cold storage at 1 °C. By functional categorization of the differentially expressed transcripts using MapMan, we found that many transcripts related to various pathways, such as jasmonic acid biosynthesis and signaling, galactinol, raffinose, phenol, and phenylpropanoid biosynthesis, calcium and mitogen-activated protein kinase signaling, lipid metabolism, and various transcription factors and heat-shock proteins, have been massively upregulated in 'Wonderful' but not in 'Ganesh' fruit. Thus, it is suggested that these pathways most likely participate in imparting chilling tolerance in pomegranate fruit.

Total anthocyanin, flavonoid, polyphenol and tannin contents of seven pomegranate cultivars grown in Iran.

BACKGROUND: Pomegranates are healthy fruits rich in bioactive phytochemicals and widely consumed in  the human diet. This study was designed to assess their physico-chemical properties and total polyphenolic compounds, along with antioxidant and ascorbic acid concentrations of the skin and juice of seven unstudied pomegranate cultivars grown in Iran. METHODS: Ten fruits of each of seven selected cultivars were picked. Methanolic extracts from the fruit skins as well as juices were subjected to an evaluation of their physico-chemical properties and phenolic content. RESULTS: From the pomegranate cultivars studied, the highest juice concentration was measured in ‘Zagh- -e-Zabol’ (72.9 ±2.55%). The highest total soluble solids (18.79 ±1.06 oBrix) of juice, as well as the highest ascorbic acid (16.54 ±2.09 mg/100 g) and total anthocyanin content of skin [11.20 ±2.4 mg CyE (cyanidin- -3-glucoside equivalent)/L] were measured in ‘Bajestan’. Total flavonoids were more concentrated in the skin of ‘Siah-e-Zabol’ [930 ±16 mg QuE (quercetin equivalent)/L]. In the juice of ‘Rabab-e-Pust Ghermez’ and ‘Malas-e-Saveh’ the highest concentrations of phenolic compounds were recorded [1082 ±12.8 mg GaE (gal- lic acid equivalent)/L] and tannins [6.55 ±0.5 mg TaE (tannic acid equivalent)/L], respectively. CONCLUSIONS: Significant differences were assessed between the skin and juice of the cultivars in terms of their physico-chemical properties and polyphenolic contents. The results showed higher levels of total tannins, phenolics and antioxidant activity in juice of pomegranate, whereas the skins possessed higher total flavo- noid, anthocyanin and ascorbic acid content. Further phytochemical analysis are therefore needed to identify the most representative phenylpropanoids by LC-MS (liquid chromatography coupled to mass spectrometry) and to develop novel and promising dietary supplements.

First reported chloroplast genome sequence of Punica granatum (cultivar Helow) from Jabal Al-Akhdar, Oman: phylogenetic comparative assortment with Lagerstroemia.

Pomegranate (Punica granatum L.) is one of the oldest known edible fruits. It has grown in popularity and is a profitable fruit crop due to its attractive features including a bright red appearance and its biological activities. Scientific exploration of the genetics and evolution of these beneficial traits has been hampered by limited genomic information. In this study, we sequenced the complete chloroplast (cp) genome of the native P. granatum (cultivar Helow) cultivated in the mountains of Jabal Al-Akhdar, Oman. The results revealed a P. granatum cp genome length of 158,630 bp, characterized by a relatively conserved structure containing 2 inverted repeat regions of 25,466 bp, an 18,686 bp small single copy regions, and an 89,015 bp large single copy region. The 86 protein-coding genes included 37 transfer RNA genes and 8 ribosomal RNA genes. Comparison of the P. granatum whole cp genome with seven Lagerstroemia species revealed an overall high degree of sequence similarity with divergence among intergenic spacers. The location, distribution, and divergence of repeat sequences and shared genes of the Punica and Lagerstroemia species were highly similar. Analyses of nucleotide substitution, insertion/deletions, and highly variable regions in these cp genomes identified potential plastid markers for taxonomic and phylogenetic studies in Myrtales. A phylogenetic study of the cp genomes and 76 shared coding regions generated similar cladograms. The complete cp genome of P. granatum will aid in taxonomical studies of the family Lythraceae.

Identification and characterization of evolutionarily conserved alternative splicing events in a mangrove genus Sonneratia.

Alternative splicing (AS), which produces multiple mRNA transcripts from a single gene, plays crucial roles in plant growth, development and environmental stress responses. Functional significances of conserved AS events among congeneric species have not been well characterized. In this study, we performed transcriptome sequencing to characterize AS events in four common species of Sonneratia, a mangrove genus excellently adaptive to intertidal zones. 7,248 to 12,623 AS events were identified in approximately 25% to 35% expressed genes in the roots of the four species. The frequency of AS events in Sonneratia was associated with genomic features, including gene expression level and intron/exon number and length. Among the four species, 1,355 evolutionarily conserved AS (ECAS) events were identified from 1,170 genes. Compared with non-ECAS events, ECAS events are of shorter length and less possibility to introduce premature stop codons (PTCs) and frameshifts. Functional annotations of the genes containing ECAS events showed that four of the 26 enriched Gene Ontology (GO) terms are involved in proton transport, signal transduction and carbon metabolism, and 60 genes from another three GO terms are implicated in responses to osmotic, oxidative and heat stresses, which may contribute to the adaptation of Sonneratia species to harsh intertidal environments.

Pomegranate Cultivars: Identification of the New IgE-Binding Protein Pommaclein and Analysis of Antioxidant Variability.

The consumption of pomegranate is increasing as it is considered a health-promoting food. Nevertheless, it can trigger allergic reactions, sometimes severe. The LTP Pun g 1 is the only pomegranate allergen so far reported. Based on preliminary clinical observations, the main aim of this study was the investigation of still unknown allergens contained in this fruit. Pommaclein, a homologue of peamaclein, the peach allergen Pru p 7, was isolated, identified by protein sequencing, and characterized as an IgE-binding protein by different test systems. RP-HPLC protein profiles revealed significant variations of LTP and pommaclein content in the red pulp of selected cultivars and accessions. Conversely, the mesocarp appeared free of proteins and much richer in antioxidants. In conclusion, a new allergen has been identified, and it could contribute to improving allergy diagnosis. The study highlights that pomegranate mesocarp could represent a rich and safe source of nutraceuticals also for allergic subjects.

Avaliação da biocompatibilidade e atividade antimicrobiana in vitro da matriz de Hidrogel associada ao extrato glicólico do Punica granatum L. (romã) / Evaluation of biocompatibility and antimicrobial action in vitro of a hydrogel scaffolds Hydromatrix associated to glycolic extract of Punica granatum (pomegranate)

A matriz de hidrogel é um biomaterial de nanofibra peptídica tridimensional que induz o crescimento, migração e proliferação celular, assim favorecendo a regeneração tecidual. O objetivo deste trabalho foi avaliar a biocompatibilidade e a atividade antimicrobiana da matriz de hidrogel associado ao extrato romã e ao antimicrobiano ciprofloxacino. Para isso, foram realizadas análises microbiológicas sobre Enterococcus faecalis (ATCC 4083) em cultura planctônica, por meio d ensaio de microdiluição em caldo e em biofilme pelo teste de MTT. Para os ensaios da biocompatibilidade, foi utilizada cultura de macrófagos (RAW 264.7). A citotoxicidade foi avaliada pelo ensaio de MTT e a genotoxicidade foi realizado pelo teste de micronúcleo. A análise estatística foi realizada pelos testes ANOVA e Tukey, adotando o nível de significância 5%. Os resultados demonstraram que a matriz de hidrogel associado ao extrato de romã não apresentou atividade antimicrobiana para a cultura planctônica como para o biofilme de E. faecalis. Porém, não foi citotóxico e genotóxico para RAW 264.7. Por outro lado, o antimicrobiano ciprofloxacino apresentou atividade antimicrobiana sobre E. faecalis,além de ter apresentado efeitos citotóxico e genotóxico para os macrófagos. Com isso, foi possível concluir que o extrato de romã associado ou não a matriz de hidrogel apresenta ausência de citotóxico, genotóxico e efeito antimicrobiano

The hydrogel matrix is a three-dimensional peptide nanofiber biomaterial that induces cell growth, migration and proliferation, thus favoring tissue regeneration. The objective of this work was to evaluate the biocompatibility and antimicrobial activity of the hydrogel matrix associated with the pomegranate extract and the antimicrobial ciprofloxacin. For this, microbiological analyzes on Enterococcus faecalis (ATCC 4083) were carried out in planktonic culture, by means of the microdilution test in broth and in biofilm by the MTT test. For the biocompatibility assays, macrophage culture (RAW 264.7) was used. Cytotoxicity was assessed by the MTT assay and genotoxicity was performed by the micronucleus test. Statistical analysis was performed by the ANOVA and Tukey tests, adopting the significance level 5%. The results showed that the hydrogel matrix associated with the pomegranate extract did not show antimicrobial activity for the planktonic culture as for the E. faecalis biofilm. However, it was not cytotoxic and genotoxic for RAW 264.7. On the other hand, the antimicrobial ciprofloxacin showed antimicrobial activity on E. faecalis, besides having cytotoxic and genotoxic effects for the macrophages. With this, it was possible to conclude that the pomegranate extract associated or not with the hydrogel matrix shows absence of cytotoxic, genotoxic and antimicrobial effect

Divergence times, historical biogeography, and shifts in speciation rates of Myrtales.

We examine the eudicot order Myrtales, a clade with strong Gondwanan representation for most of its families. Although previous phylogenetic studies greatly improved our understanding of intergeneric and interspecific relationships within the order, our understanding of inter-familial relationships still remains unresolved; hence, we also lack a robust time-calibrated chronogram to address hypotheses (e.g., biogeography and diversification rates) that have implicit time assumptions. Six loci (rbcL, ndhF, matK, matR, 18S, and 26S) were amplified and sequenced for 102 taxa across Myrtales for phylogenetic reconstruction and ten fossil priors were utilized to produce a chronogram in BEAST. Combretaceae is identified as the sister clade to all remaining families with moderate support, and within the latter clade, two strongly supported groups are seen: (1) Onagraceae+Lythraceae, and (2) Melastomataceae+the Crypteroniaceae, Alzateaceae, Penaeaceae clade along with Myrtaceae+Vochysiaceae. Divergence time estimates suggest Myrtales diverged from Geraniales ∼124Mya during the Aptian of the Early Cretaceous. The crown date for Myrtales is estimated at ∼116Mya (Albian-Aptian). BioGeoBEARS showed significant improvement in the likelihood score when the "jump dispersal" parameter was added. South America and/or Africa are implicated as important ancestral areas in all deeper nodes. BAMM analyses indicate that the best configuration included three significant shifts in diversification rates within Myrtales: near the crown of Melastomataceae (∼67-64Mya), along the stem of subfamily Myrtoideae (Myrtaceae; ∼75Mya), and along the stem of tribe Combreteae (Combretaceae; ∼50-45Mya). Issues with conducting diversification analyses more generally are examined in the context of scale, taxon sampling, and larger sets of phylogenetic trees.

Inhibitory Effects of Two Varieties of Tunisian Pomegranate (Punica granatum L.) Extracts on Gastrointestinal Transit in Rat.

The present study was undertaken to determine whether total and methanol juice extracts of two Tunisian Pomegranate (Punica granatum L.) varieties (Garsi and Gabsi) protect against diarrhea as well as their effects on gastrointestinal transit (GIT) in healthy rats. In this respect, male Wistar rats were used and divided into control- and pomegranate-treated groups. The antidiarrheal activity was evaluated using the castor oil-induced diarrhea method and the GIT was assessed using charcoal meal. Our results showed that total and methanol P. granatum juice extracts produced a significant dose-dependent protection against castor oil-induced diarrhea. Pomegranate extracts and juice also decreased the GIT significantly and dose dependently. Importantly, the Garsi variety appeared to be more effective than the Gabsi variety on these two parameters. These findings suggest that pomegranate extracts have a potent antidiarrheal property in rats confirming their efficiency in the Tunisian traditional medicine.

Genetic diversity and association mapping of bacterial blight and other horticulturally important traits with microsatellite markers in pomegranate from India.

This genetic diversity study aimed to estimate the population structure and explore the use of association mapping strategies to identify linked markers for bacterial resistance, growth and fruit quality in pomegranate collections from India. In total, 88 accessions including 37 cultivated types were investigated. A total of 112 alleles were amplified by use of 44 publicly available microsatellites for estimating molecular genetic diversity and population structure. Neighbor-joining analysis, model-based population structure and principal component analysis corroborated the genetic relationships among wild-type and cultivated pomegranate collections from India. Our study placed all 88 germplasm into four clusters. We identified a cultivated clade of pomegranates in close proximity to Daru types of wild-type pomegranates that grow naturally near the foothills of the Himalayas. Admixture analysis sorted various lineages of cultivated pomegranates to their respective ancestral forms. We identified four linked markers for fruit weight, titratable acidity and bacterial blight severity. PGCT001 was found associated with both fruit weight and bacterial blight, and the association with fruit weight during both seasons analyzed was significant after Bonferroni correction. This research demonstrates effectiveness of microsatellites to resolve population structure among the wild and cultivar collection of pomegranates and future use for association mapping studies.

Antioxidant and lipase inhibitory activities and essential oil composition of pomegranate peel extracts.

The chemical composition of essential oil, antioxidant and pancreatic lipase inhibitory activities of various solvent extracts obtained from pomegranate peelTunisian cultivar was evaluated. Gas chromatography/mass spectrometry was used to determine the composition of the PP essential oil. Nine-teen components were identified and the main compounds were the camphor (60.32%) and the benzaldehyde (20.98%). The phenolic and flavonoids content varied from 0 to 290.10 mg Gallic acid equivalent and from 5.2 to 20.43 mg catechin equivalent/g dried extract. The antioxidant activity of various solvent extracts from pomegranate peel was also investigated using various in vitro assays as the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical method, ß-carotene bleaching and reducing power assays.Methanol and ethanol extracts showed the most potent antioxidant activity in all assays tested followed by water and acetone extracts. The inhibitory effect of the pomegranate peelextracts on porcine pancreatic lipase was evaluated and the results showed that ethanol and methanol extracts markedly reduced lipase activity. Generally, the highestlipase activity inhibitory (100%) was observed at a concentration of 1 mg/ml after 30 min of incubation. LC-MS analysis of ethanol extract showed the presence of four components which are cholorogenic acid, mannogalloylhexoside, gallic acid and ellagic acid. Our findings demonstrate that the ethanol extract from pomegranate peel might be a good candidate for furtherinvestigations of new bioactive substances.

Studying genetic variability of pomegranate (Punica granatum L.) based on chloroplast DNA and barcode genes.

Chloroplast DNA has been used extensively to analyze plant phylogenies at different taxonomic levels because of its size, organization and sequence conservation. In the present research, two chloroplastic regions, petA­psaJ, trnC­trnD and four DNA barcodes (trnH­psbA, ITS, rbcL, matK), were used to introduce suitable regions for the assessment of genetic diversity among P. granatum L. genotypes. Analysis of psbE­petL in petA­psaJ region revealed 1,300 nucleotides with 4.29 % genetic diversity among genotypes, while trnC­petN in trnC­trnD region showed 1.8 % genetic diversity. Therefore, despite the results obtained from the study of other plants, the trnC­trnD region had a low potential for the evaluation of diversity among pomegranate genotypes. Analysis of DNA barcodes in pomegranate showed that trnH­psbA (genetic diversity 2.91 %) provides the highest intra-species variation, followed by ITS (genetic diversity 0.44 %). Eighteen genotypes from different geographical origins of Iran were used to investigate psbE­petL and trnH­psbA potential as novel barcodes to determine genetic polymorphism and characterize pomegranate genotypes. The results suggested that two regions, psbE­petL and trnH­psbA, were more suitable for determining intra-species relationships of pomegranate.

Chemical composition and antioxidant, anti-inflammatory, and antiproliferation activities of pomegranate (Punica granatum) flowers.

The chemical composition, antioxidant (DPPH and ABTS assays), anti-inflammatory (5-LOX), and cytotoxic (MCF-7) activities from flowers of seven pomegranate varieties (Punica granatum) were investigated. The highest phenolics (330.9±11.3 mg gallic acid equivalent/g dry weight (dw)), flavonoids (29.5±0.8 mg quercetin equivalent/g dw), tannins (30.6±0.6 mg catechin equivalent/g dw), and anthocyanins (0.70±0.03 mg cyanidin-3-glucoside equivalent/g dw) content were determined in the Chetoui (CH) variety. It was found that Garsi (GR) (IC50=4.9±0.2 mg/L by ABTS assay) and Zaghwani (ZG) (IC50=3.9±0.2 mg/L by ABTS assay) varieties exhibited the highest antioxidant activity. For the anti-inflammatory activity, all varieties were active; the ZH variety was the strongest (2.5±0.1 mg/L). The CH, ES, and RA pomegranate varieties were not active against human breast cancer cells MCF-7, whereas inhibition was more evident with extracts from ZH and GR varieties (IC50=33.00±2.64 and 35.00±4.58 mg/L, respectively). Statistical analysis showed that the variety factor influenced significantly (P<.01) the chemical composition and biological activities of pomegranate flowers.

A novel strategy for identification of 47 pomegranate (Punica granatum) cultivars using RAPD markers.

DNA marker can be used for precise plant cultivar identification. However, DNA markers have often not been used effectively for the identification of plant cultivars due to a lack of an effective analysis strategy. We used a novel strategy for effective identification of plant individuals based on a new way of recording DNA fingerprints of the genotyped plants; a cultivar identification diagram can be manually generated and used as key reference information for quick identification of plant and/or seed samples. Forty-seven pomegranate varieties popularly cultivated in various provinces of China were subjected to RAPD marker analysis. Using the cultivar identification diagram strategy, they were clearly separated by the fingerprints of 11 RAPD primers. The utility and accuracy of the cultivar identification diagram analysis results were confirmed by the identification of three randomly chosen groups of cultivars among the 47 varieties.

Biochemical analysis of two varieties of water chestnuts (Trapa sp.).

In this study, two varieties (Green and red) of water chestnuts (Trapa sp.) have been selected for their biochemical analysis as well as nutrient composition using standard methods. The proximate composition of green water chestnuts revealed moisture 62.5, ash 1.04, crude fiber 2.13%, total soluble sugar 0.92%, reducing sugar 0.33%, non-reducing sugar 0.59%, starch 8.7%, lipid 0.84%. One hundred gram of green variety contained water soluble protein 0.275 mg, beta-Carotene 60 microg, vitamin-C 1.1 mg and total phenol 0.5 mg. The minerals contents of green variety were potassium 5.22%, sodium 0.64%, calcium 0.25%, phosphorus 6.77%, sulpher 0.38%, and iron, copper, manganese and zinc 200, 430, 90 and 600 ppm, respectively. The red variety contained moisture 62.7%, ash 1.30%, crude fiber 2.27%, total soluble sugar 0.90%, reducing sugar 0.30%, non-reducing sugar 0.60%, starch 8.2%, lipid 0.83%. The red variety contained water soluble protein 0.251 mg, beta-Carotene 92 microg, vitamin-C 0.9 mg and total phenol 0.60 mg per 100 g. The red variety contained potassium 5.32%, sodium 0.59%, calcium 0.26% phosphorus 6.77%, sulpher 0.32%, Iron 200 ppm, copper 450 ppm, manganese 110 ppm and zinc 650 ppm. The free amino acids, glutamic acid, tryptophan, tyrosine, alanine, lysine and leucine were commonly found in both varieties. In addition, green and red variety contained cysteine, arginine and proline and glutamine and asparagines, respectively. Thus, the present study sheds light on the nutrient contents of the two varieties of water chestnuts and suggests that water chestnuts may play a crucial role in human nutrition.

Lythrum and Peplis from the Late Cretaceous and Cenozoic of North America and Eurasia: new evidence suggesting early diversification within the Lythraceae.

PREMISE OF THE STUDY: To fully understand the evolution of today's angiosperms, the fossil record of plant families and genera must be used to determine their time of origin and phytogeographic history. As within many angiosperm families, the interrelationships of extant Lythraceae are hard to resolve without sufficient data from the geological past. Here we establish the earliest fossil occurrences of Lythraceae and start resolving the interrelationships and evolution of two of its genera, Lythrum and Peplis. METHODS: We studied several Cretaceous and Cenozoic palynofloras from the northern and southern hemispheres. Using the single-grain technique, we screened the treated samples for Lythrum- and Peplis-type pollen. The same individual pollen grains were observed under both the light- and scanning electron microscope, allowing a high taxonomic resolution to be achieved. KEY RESULTS: Fossil Lythraceae pollen grains are rare in palynological samples. Nevertheless, we were able to identify Lythrum and Peplis pollen from Late Cretaceous sediments and thereby extend the fossil record of the two genera by ca 70 million years. CONCLUSIONS: The appearance of Lythrum and Peplis in North America and Peplis in Asia at approximately the same interval in the mid Late Cretaceous points to an already wide geographical distribution by then. These findings add vital information for the time of origin of the Lythraceae and suggest a higher diversity within the family. They also indicate that the distribution of particular genera during the Cretaceous was wider than previously thought.