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Coix seed oil (CSO) has a wide range of anticancer effects. However, the mechanism of action against pancreatic cancer (PC) and regulation of mitochondria in vitro is still unclear.This research investigated the possible mechanism of CSO induction of PC cell apoptosis and regulating mitochondrial functional damage. Proliferation of PC cells, mitochondrial membrane potential (MMP), qualitative and quantitative analysis of PC cell apoptosis, openness of mitochondrial permeability transition pore, related protein expression, generation of reactive oxygen species (ROS), and gene expression were determined by cell counting kit-8, JC-1 staining, acridine orange and ethidium bromide staining, flow cytometry, calcein-AM/cobalt staining, western blotting, dichlorofluorescein diacetate probe, and quantitative real-time reverse transcription-polymerase chain reaction, respectively. We confirmed that PTEN protein was involved in CSO-induced PANC-1 cell apoptosis and mitochondrial functional damage. CSO induced depolarization of MMP, increased opening of the mitochondrial permeability transition pore, increased ROS production, and further increased mitochondrial damage. Additionally, CSO downregulated expression of p-AKT and p-PI3K proteins; upregulated protein expression of cleaved caspase-9, Bax, cleaved caspase-3 and cytochrome c; and downregulated expression of Bcl-2 by upregulating the PTEN gene. The corresponding protein expression was consistent with the gene expression level. Furthermore, the loss of function of PTEN protein reduces the ability of CSO to induce apoptosis of PANC-1 cells and damage to mitochondrial function.CSO induces apoptosis of PANC-1 PC cells by modulating mitochondrial functional impairment and related apoptotic molecules via PTEN, which may be closely related to the PI3K/AKT signaling pathway.

Sanghuangporus sanghuang (SS) is a rare medicinal polypore fungus that grows solely on Morus trees. In this study, seven grains (oats, barley, millet, rice, buckwheat, corn, and coix seed) were used as solid substrates for SS fermentation and characterized in their nutrition, functional composition, and antioxidant activities. After fermentation, the nutrient compositions of crude protein (F1,41 = 111.1, P F1,41 = 595.7, P F1,41 = 51.4, P F1,41 = 227.3, P ΔT-AOC/ABTS+/DPPH/DNAp, that was correlated to part of changes in polyphenol, carotenoid, triterpenoids, and flavonoid contents. In summary, oats have the greatest potential for use as a fermentation substrate for health food development.

Coix lacryma-jobi L. is an Asian plant widely used in traditional medicine. Drought conditions can lead to an impairment of photosynthesis which can be softened by the exogenous application of proline and γ-aminobutyric acid (GABA) to maintain the electron transport chain (ETC) and avoid reactive oxygen species-induced oxidative damage by inducing antioxidative enzymes. The aim of this study was to evaluate the effects of water deficit on C. lacryma-jobi leaves under the application of proline and GABA. Chlorophyll (Chl) a fluorescence emission, antioxidative enzyme activity (catalase, superoxide dismutase, ascorbate peroxidase and guaiacol peroxidase) and proline and GABA contents in leaf tissues were performed in plants subjected to 12 days of drought. Chl a fluorescence indicated impairments on photosynthetic apparatus due to water deficit that resulted into increased activity of antioxidant enzymes. The application of proline but not GABA was more efficient in alleviating the effects of water deficit, as evidenced by the maintenance of the dynamic dissipation of the photosynthetic energy, besides the greater accumulation of proline under these conditions. Water deficit induces alterations in the ETC, which is softened by the application of proline to cope with the deleterious effects of drought in C. lacryma-jobi plant leaves.

Wound dressings are based on the creation of ideal environments for cell mobility, gas exchanges and to promote tissue healing and regeneration, besides controlling bacterial proliferation. Electrospinning is a simple, cheap and common technique used for such purposes and Ecovio®, a polymeric blend based on biocompatible polymers (poly(butylene adipate-co-terephthalate) (PBAT) and poly(lactic acid) (PLA)), is a good example of polymer with unique properties for wound dressing materials development. To ensure antimicrobial properties, Job’s tears (Coix lacrymajobi) extract was incorporated into the material. The obtained fiber mats were characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffractometry (DRX), optical microscopy (OM), and microbiological analysis. The interaction among the compounds of the extract and the polymeric matrix was inferred by the increase in the crystallinity degree and thermal stability of the fibers/extract. Also, a decrease in the diameter, roughness, and fiber homogeneity was observed. It happened due to the presence of extract in the fibers. Microbiological analysis (antimicrobial activity against S. aureus) indicated the effectiveness of the fibers/extract in preventing this bacteria growth, demonstrating the potential of this material for wound dressing purposes.

A traditional Chinese medicine formula based on the Coix seed and Lotus seed has been used as a general treatment of malnutrition, excessive fatigue, dysfunction of the spleen and stomach, and disorders of water transport in humans in China. However, there is limited information on its effects on the gut microbiota of piglets in vivo. In this study, the mix of Coix seed and Lotus were added the diet of forty weaned piglets (local piglets), and then evaluated it’s affected on the gut microbiota of piglets and on the relations within the gut bacterial community. The results indicated that this traditional Chinese medicine formula (LM) and the extract of the traditional Chinese medicine formula (LMT) downregulated pH of succus gastricus and raised pH of the ileum, and LMT obviously decreased the feed conversion ratio. Further study showed LMT and LM also significantly increased the thick and long of gastrointestinal villi. And then, 16S ribosomal DNA sequencing revealed that groups LMT and LM have higher relative abundance of the genus Lactobacillus in the colon, succus gastricus, and jejunum, which are beneficial bacteria sold as dietary supplements to aid digestion or to augment health. Meanwhile, the relative abundance levels of Prevotellaceae, Alloprevotella, and Prevotella in the colon and Clostridium in succus gastricus and jejunum were lower. These experiments highlight the usefulness of the traditional Chinese medicine formula based on the Coix seed and Lotus seed for decreasing pH in succus gastricus, for improving the structure of intestinal villi and gut microflora, and then for achieving improvements in pig production performance.

The founding processes of the first state of ancient China with a known written record, the Shang dynasty (3600–3046 cal BP), have been poorly understood. Recent discoveries of a host of archaeological sites dating to the proto-Shang culture (4000–3600 cal BP) have helped elucidate the transition to the Shang culture. Nevertheless, there are few investigations about the mode of subsistence and economy of the proto-Shang culture, and how this might have shaped the transition to statehood. In this present study, we analyzed the starch grains preserved in dental calculus and teeth surfaces from 16 samples from the site of Nancheng in order to gain a better understanding of the subsistence strategy and plant consumption of proto-Shang people. We also performed experiments to test how different cooking methods may lead to size changes in the starches of four Poaceae plants, in order to identify the processing methods used by the proto-Shang people. The results indicate that Triticum aestivum, Coix lacryma-jobi, Setaria italica and some yet-unidentified roots and tubers were consumed by these individuals. These data indicate a broader spectrum of plant consumption than that seen by previous archaeobotanical and stable isotope analyses. Such a broad spectrum of plant consumption provided a substantial economic base for proto-Shang people and might be one of the factors supporting the subsequent development of the Shang state culture.

Job’s tear grain (Coix lachryma-jobi L.) was hydro-thermally treated to study the effects of heat-moisture treatment on the milling, nutritional and certain physicochemical properties of the grain. The milling yield improved by 30% after processing due to hardening of the grain. This was proved by the texture profile analysis which showed the increased trend in hardness with the increase in the intensity of heat treatment on the grains. The total starch content of the grain was 75–82 g 100 g−1, with just 2–5 g 100 g−1 being amylose. Soaking before steaming has a greater effect on water absorption capacity and solubility power of the grain; it also improved cooking time. The pasting and thermal properties of native and steamed grains were similar but there was a difference in soak and steamed grains. The presence of non-starch components like protein and lipids also played a role in gelatinization and viscosity of the grain.

Pain is one of the most commonly reported symptoms in patients with advanced cancer, but is still less than optimally treated. The effect of traditional Chinese medicine in cancer pain treatment is nowadays getting more and more attention. To investigate the effect of a coix seed oil injection on cancer pain relief in a cancer center in a tertiary hospital in China. Patients in the treatment group received a coix seed oil injection for 2 weeks, while patients in the control group received equivalent 0.9% saline. The numeric rating scale was used to assess the pain level. The Quality of Life Questionnaire-Core 30 was used to assess life quality. The adverse drug reactions during the treatment process were observed. Patients in the coix seed treatment group had significantly superior efficacy on pain control over those in the control group. Coix seed therapy significantly improved patients’ scores reflecting by the Quality of Life Questionnaire-Core 30 (QLQ-C30) scale. In addition, the occurrence of adverse reactions such as constipation and nausea in the treatment group was significantly lower than that in the control group. The coix seed oil injection effectively reduced the pain level of cancer patients, significantly improved their life quality, and had no obvious adverse effects.

Background Cancer cachexia is a cancer-induced multifactorial debilitating syndrome directly accounting for 20% of cancer deaths without effective therapeutic approaches. It is extremely urgent to explore effective anti-cachexia drugs to ameliorate muscle and fat loss in cachexia patients. Methods Lewis lung carcinoma bearing C57BL/6 mice were applied as the animal model to examine the therapeutic effect of Coix seed oil (CSO) on cancer cachexia. The food intake and body weight change were monitored every 3 days throughout the experiment. The IL-6 and TNF-α levels in serum were detected by ELISA assay. Several key proteins involved in muscle wasting and fat lipolysis were tested by Western blot to identify the potential mechanism of CSO. Results Administration of CSO through gavage significantly prevented body weight loss and ameliorated systemic inflammation without affecting food intake and tumor size. The weight and histological morphology of gastrocnemius muscle and epididymal adipose tissue in CSO-treated mice were also improved. In mechanism, we found that CSO decreased the expression of MuRF1 and the ratio of phospho-p65 (Ser536) to p65 in muscle tissue. Meanwhile, cancer-induced activation of HSL and AMPK was also inhibited by CSO administration. Conclusion Coix seed oil exerts an anti-cachexia pharmaceutical effect by counteracting muscle and adipose tissue loss most likely through regulating NF-κB-MuRF1 and AMPK-HSL pathway.

Coix seed has traditionally been used in traditional Chinese medicine to fortify the spleen and inhibit dampness, and has shown anticancer effects in humans. However, it is not known whether coix seed improves post-weaning growth performance and productivity, and the mechanism of interaction between coix seed and gut microbiota remains unknown. In this study, we established four groups: (i) control, (ii) antibiotic-fed, (iii) coix seed powder-fed, and (iv) coix seed extract-fed. The feeding experiment was conducted for 4 weeks. Coix seed extract significantly increased average weight gain and reduced the feed/meat ratio in weaned pigs, in addition to reducing the pH of their gastric juice. Further assays demonstrated that coix seed promotes an increase in the density and length of the gastrointestinal villi. Next, 16s sequencing of gut microbiota showed that coix seed significantly increased the abundance of phylum Bacteroidetes and genus Lactobacillus (p

Nanostructure with an interior nanogap has received much attention in surface-enhanced Raman scattering (SERS). However, controllable synthesis of nanostructure with ultrasmall nanogap and innovative morphology still remains a challenge. Herein, we present a facile seed-mediated route to integrate uniform nanogap in novel Ag nano coix seeds and locate Raman molecules in the nanogap at room temperature. After 20-nm Ag nanoparticles (NPs) modified by Raman ligand 2-naphthalenethiol and coated by polymer shells as cores were obtained, outside Ag shells were formed by in situ reduction on the polymer surface. The SERS properties of these resulting Ag nano coix seeds were systematically explored. More importantly, the novel SERS active substrate exhibited ultrahigh homogeneity, reproducibility, stability, and even a reliable quantitative SERS analysis mediated by internal standard molecules.

Magnetic solid-phase extraction (m-SPE) is a promising sample preparation approach due to its convenience, speed, and simplicity. For the first time, a rapid and reliable m-SPE approach using magnetic multi-walled carbon nanotubes (m-MWCNTs) as the adsorbent was proposed for purification of type A trichothecenes including T-2 toxins (T2), HT-2 toxins (HT-2), diacetoxyscirpenol (DAS), and neosolaniol (NEO) in coix seed. The m-MWCNTs were synthesized by assembling the magnetic nanoparticles (Fe3O4) with MWCNTs by sonication through an aggregation wrap mechanism, and characterized by transmission electron microscope. Several key parameters affecting the performance of the procedure were extensively investigated including extraction solutions, desorption solvents, and m-MWCNT amounts. Under the optimal sample preparation conditions followed by analysis with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), high sensitivity (limit of quantification in the range of 0.3-1.5 μg kg(-1)), good linearity (R (2) 0.99), satisfactory recovery (73.6-90.6 %), and acceptable precision (≤2.5 %) were obtained. The analytical performance of the developed method has also been successfully evaluated in real coix seed samples. Graphical Abstract Flow chart of determination of type A trichothecenes in coix seed by magnetic solid-phase extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry.

The upper-middle Huai River valley (HRV) is located in the transition zone in the middle of eastern China. Previous studies have shown that the past populations in the upper HRV obtained plant food through gathering and cultivation of both rice and millet during the middle Neolithic. However, for the middle HRV it is not very clear what methods were used by humans to obtain plant food in that time. In this paper, starch grain analysis was carried out on 17 stone tools and 29 cauldron pottery sherds unearthed at Shunshanji (8.5–7.0 kyr BP), which is known as the earliest Neolithic site in the middle HRV excavated so far. Here, ancient starches from Coix lacryma-jobi, Triticeae, Oryza sativa, Trichosanthes kirilowii and one unidentified specie were recovered. This study contributes to the limited knowledge of food strategies as observed in the middle HRV. It demonstrates that both gathering and cultivation of rice took place during the middle-Neolithic. Among the identified species, Coix lacryma-jobi appears to have been the main plant food at Shunshanji. Moreover, it is clear that Oryza sativa was not consumed as much as other plant species according to the occurrence frequency of different starch grains. Starch grains were also found on the used surfaces of grinding stone implements as well as pestles, which means that these stone tools were used for food processing. In addition, axes from Shunshanji may also have been used for food processing as well as wood working because starches were also found on the edge of axes. Information about subsistence strategies and tool use at Shunshanji will also be helpful to understanding the utilization of plants and agricultural development in the middle HRV during the early-middle Neolithic period.

Background It is well known that endoplasmic reticulum (ER) stress plays a huge role in development of metabolic diseases. Specially, ER stress-induced cellular dysfunction has a significant involvement in the pathogenesis of human chronic disorders. This study was designed to study to assess whether an ethanol extract of Coicis Semen (CSE) and coixol induces the ER stress in Chang liver cells. Methods Coicis Semen was mixed with 95% ethanol at a ratio of 1:10 (w/v) and freeze dried. Chang liver cells were seeded to 96-well plates and treated with or without CSE (100, 200, 300, 500, or 1000 μg/mL) or coixol (100, 200, 300, 500, 750, or 1000 μg/mL). cell viability was analyzed with MTT assay. Effects of CSE and coixol on expression of the genes for ER stress markers were determined with qRT-PCR and the expression of the protein levels of ER stress markers were determined with western blotting. Results The concentration causing 50% inhibition (IC50) for CSE and coixol was 250 and 350 μg/mL, respectively. The CSE and coixol increased the gene expression of BiP and CHOP in a dose-dependent manner. Furthermore, CSE and coixol dose-dependently increased the the expression of XBP1. Conclusions CSE or coixol may have cytotoxic effect to Chang liver cells and, may induce ER stress and stimulate the UPR via activation of the PERK and IRE1 pathways in normal liver cells. Electronic supplementary material The online version of this article (10.1186/s12906-018-2175-z) contains supplementary material, which is available to authorized users.

The seed of the Coix lacryma-jobi L. var. ma-yuen Stapf seed (CLMS) extract has been prescribed to alleviate obesity by practitioners of traditional Korean medicine. Here, we investigated the effect of CLMS extract on PPARγ 2 and c/EBPα and obesity responses in C57BL/6J obese mice fed on a high fat diet. The mouse body index, blood profile, and fat accumulation levels in the liver were measured. The protein expression levels of PPARγ 2 and c/EBPα in the mice livers were also measured to determine the molecular mode-of-action of the reducing effect of CLMS extract on mouse adipogenesis. The results showed that HFD-induced mouse obesity, fat accumulation, and serum cholesterol were alleviated by the CLMS extract addition. Moreover, PPARγ 2 and C/EBPα, proteins, those are related to the adipogenesis, were downregulated by the CLMS extract intake considerably. This study indicates that as a food additive, CLMS extract has a reducing effect on the high-fat diet induced fat accumulation and on body weight through the downregulation of adipogenesis.

The feasibility of near-infrared (NIR) spectroscopy for determining total dietary fiber (TDF) and mineral elements (mainly K, Mg, P, and S) in Coix seed was investigated. Partial least squares regression (PLSR) was applied to establish quantitative models. Norris derivative smoothing (NDS) was used as pretreatment method. A comparison of three variable selection methods, namely competitive adaptive reweighted sampling (CARS), genetic algorithms (GA), and random frog (RF), showed that CARS obtained the best performances of PLSR models with the effective wavelengths mainly concentrated on around 12,000–11,000 cm−1 and 6500–3600 cm−1. For the quantitative determination models of TDF, K, Mg, P, and S, the optimal root mean square error of prediction (RMSEP) values were 0.0923, 182.7224, 75.4987, 162.6993, and 36.6278; the r values were 0.95, 0.88, 0.80, 0.96, and 0.96; the residual predictive deviation (RPD) values were 2.68, 2.05, 1.70, 3.24, and 3.04, respectively. It is concluded that the NIR spectral technique has a potential to determine TDF, K, Mg, P, and S in Coix seed.

The feasibility of near-infrared (NIR) spectroscopy to determine fat, protein, and amino acids (AAs) in Coix seed was investigated. Partial least squares (PLS) regression was applied to establish quantitative model. Competitive adaptive reweighted sampling (CARS) and Norris derivative smoothing (NDS) were used to improve the model accuracy. For fat, protein, Asx, threonine, serine, Glx, alanine, leucine, proline, lysine, and histidine, NDS pretreatment improved the models’ performance and yielded better prediction results. Then, key variables were selected by CARS, and the PLS models could obtain the optimal results with good predictive ability and robustness. The models of protein, Asx, serine, Glx, alanine, valine, isoleucine, leucine, phenylalanine, and proline were fit for screening with the residual predictive deviation (RPD, the ratio between the standard deviation of the reference value and the root mean square error of prediction of the validation set) equal or greater than 2.50. The RPDs of fat and threonine were 1.61 and 2.00, and the other AAs’ were less than 1.50. It was concluded that the NIR spectral technique was suitable for determining fat, protein, and most of AAs in Coix seed. The technique might also give a rough estimate of the contents of glycine, cysteine, methionine, tyrosine, arginine, lysine, and histidine.

In this study, the isolation of an endophytic fungus from the leaves of the medicinal herb adlay (Coix lacryma-jobi L. var. ma-yuen Stapf) is reported for the first time. The fungus produced Triolein (trioleoylglycerol), a major constituent of triacylglycerols (TAGs) of adlay, in rice medium under shake-flask and bench-scale fermentation conditions. The fungus was identified as Gibberella moniliformis (Fusarium verticillioides) by its morphology and authenticated by ITS analysis (ITS1 and ITS2 regions and the intervening 5.8S rDNA region). Triolein was identified by HPLC-ELSD coupled with APCI-MS and confirmed through comparison with authentic standard. The concentration of triolein produced by G. moniliformis AH13 reached 2.536 ± 0.006 mg/g dry weight of mycelium. Moreover, the EtOAc extract of G. moniliformis AH13 showed strong antitumor activity against four types of tumor cells (A549, HCT116, MDA-MB-231, and SW1990). These results suggest that G. moniliformis AH13 in adlay has significant scientific and industrial potential to meet the pharmaceutical demands and sustainable energy requirements for TAGs in a cost-effective, easily accessible, and reproducible way and is also a potential novel source of natural antitumor bioactive agents.

In this study, we investigated whether natural products including coixol derived from Coix Lachryma-Jobi var. ma-yuen affect MUC5AC mucin gene expression, production and secretion from airway epithelial cells. Confluent NCI-H292 cells were pretreated with oleic acid, linoleic acid, glyceryl trilinoleate, beta-stigmasterol or coixol for 30 min and then stimulated with PMA (phorbol 12-myristate 13-acetate), EGF (epidermal growth factor) or TNF-α (tumor necrosis factor-α) for 24 h. The MUC5AC mucin gene expression, mucin protein production and secretion were measured by RT-PCR and ELISA. The results were as follows: (1) Oleic acid, linoleic acid, glyceryl trilinoleate, beta-stigmasterol and coixol inhibited the expression of MUC5AC mucin gene induced by PMA from NCI-H292 cells; (2) Oleic acid, linoleic acid, glyceryl trilinoleate, beta-stigmasterol and coixol also inhibited the production of MUC5AC mucin protein induced by the same inducers from NCI-H292 cells; (3) Coixol inhibited the expression of MUC5AC mucin gene and production of MUC5AC mucin protein, induced by EGF or TNF-α from NCI-H292 cells; (4) Coixol decreased PMA-induced MUC5AC mucin secretion from NCI-H292 cells. This result suggests that coixol, the characteristic component among the examined five natural products derived from C. Lachryma-Jobi var. ma-yuen, can regulate gene expression, production and secretion of mucin, by directly acting on airway epithelial cells.

To investigate the efficacy of dietary therapy of qi-yin-reinforcing porridge for the alleviation of clinical symptoms during FOLFOX chemotherapy.A single-case randomized controlled study was carried out. Fourteen patients with gastrointestinal tumors accepting FOLFOX chemotherapy were enrolled using self-crossover control design. On days 1-7 of chemotherapy, the dietary therapy experimental group and the placebo control group were given dietary therapy of qi-yin-reinforcing porridge and dextrin-prepared porridge which had the same appearance, smell, color and taste as the qi-yin-reinforcing porridge, respectively. Fourteen clinical symptoms, including debility, vomiting and nausea, etc. were observed.Ten patients completed the study and were estimable. The qi-yin-reinforcing porridge demonstrated certain efficacy in alleviating clinical symptoms of patients with gastrointestinal tumors during the period of chemotherapy. Better effect of alleviating debility was found in the dietary therapy experimental group than in the placebo control group, and the difference was statistically significant (Z=2.27, P=0.02). No statistically significant difference was found between the experimental group and the control group with respect to the effect of alleviating the other 13 clinical symptoms including anorexia, nausea, vomiting, dry stool, loose stool, etc. and body weight (P0.05).Dietary therapy of qi-yin-reinforcing porridge shows an additional benefit in alleviating debility of patients during chemotherapy and deserves clinical use and popularization.

Plant utilization is poorly known in South China as compared to North China. Fanchengdui, Zhuweicheng, Shinianshan, and Yinjiaping, which date back to 5-4 ka BP, are typical Neolithic sites located at the middle reaches of the Ganjiang River in Jiangxi Province. Due to the lack of systematic plant archaeological work, only rice straw and husk residues have been found in some archaeological sites in Jiangxi Province. Thirteen stone tools excavated from these sites were examined in this study. The results showed that among the plants utilized are Coix spp., Oryza spp., Vigna spp., plants from the tribe Triticeae, roots from some plants of Zingiberaceae, and other species of roots and tubers. More than 457 grains were retrieved from the 13 stone tools. The fact that Coix spp. were found in all stone tools suggests that these species were a major food source during the Neolithic era. In addition, 28 starch grains of Oryza spp. were found in 8 stone tools. The analysis also revealed that not only seeds of Gramineae but also those of Leguminosae, Zingiberaceae and other species of roots and tubers are among the plants utilized in the four Fanchengdui culture sites. These results indicate that ancient plant utilization was rather diverse, and this study elucidates the prehistoric agricultural systems of South China.

We isolated the entire sequence of the coding region of Waxy gene of a non-waxy accession of Job’s tears (Coix lacryma-jobi) by PCR-based methods. We also compared the entire sequences of the gene between two non-waxy accessions and three waxy cultivars and found a 275-bp deletion in the coding region (exons 10–11) of this gene specific to waxy cultivars. We showed by PCR genotyping that this deletion is commonly found in Japanese and Korean cultivars and confirmed that this deletion resulted in lack of Wx protein. We also confirmed that this polymorphism of the gene co-segregates with phenotypes in endosperm and pollen. These results suggest that this PCR-based marker will be useful in breeding of Job’s tears and that genetic information obtained in other grass species will be also useful in genetics and breeding of Job’s tears.

Fructooligosaccharide (FOS), a prebiotic was extracted from the grain of Coix lachryma-jobi Linn. (Job's tears) by hot water extraction at 60 °C for 1 h. The resulting dried powder extract was assayed for FOS content of 1-kestose (GF2), nystose (GF3) and 1-β-D-fructofuranosylnystose (GF4) using HPLC equipped with RI detector. Total FOS content of the extract was 24.98 ± 7.48% (g/100 g crude extract). The biological activity including antioxidant and cytotoxicity of the FOS-containing extract was determined. The antioxidant activity by DPPH free radical scavenging of FOS-containing extract was comparable to vitamin C (0.97 fold of vitamin C) with a slight lipid peroxidation inhibition activity. The extract exhibited no cytotoxic effect on normal human skin fibroblast. These results have confirmed not only the source of FOS from Job's tears extract but also its potential application as antioxidant in food or cosmetic products.

Among a natural population of Coix aquatica Roxb., a pentasomic (2n + 3; 2n = 15) was isolated and studied cytologically for the first time. The extra homologous chromosomes in the plant behaved normally, as they did not disturb the rest of the bivalents through meiosis except for the stickiness and rare occurrence of a few univalents and chromosome fragments. The majority of pollen mother cells (PMCs) showed five bivalents and a pentavalent at diakinesis. The different anaphase segregations led to the formation of hypo- and hyper-ploid male gametes from n-1 (n = 5) to n + 4 (n = 10) chromosomal constitutions. Although the chromosomal configurations towards the later stages were found to be sticky in most of the meiocytes, the meiosis was clean, giving rise to pollens that appeared to be normal. The plant yielded a few fertile seeds also. The origin of this higher polysomic is considered to be through the fusion of male and female super haploid gametes produced by the trisomics that arose from the 2n = 12 Coix taxon.

Tandemly arrayed genes (TAGs) account for about one-third of the duplicated genes in eukaryotic genomes. They provide raw genetic material for biological evolution, and play important roles in genome evolution. The 22-kDa prolamin genes in cereal genomes represent typical TAG organization, and provide the good material to investigate gene amplification of TAGs in closely related grass genomes. Here, we isolated and sequenced the Coix 22-kDa prolamin (coixin) gene cluster (283 kb), and carried out a comparative analysis with orthologous 22-kDa prolamin gene clusters from maize and sorghum. The 22-kDa prolamin gene clusters descended from orthologous ancestor genes, but underwent independent gene amplification paths after the separation of these species, therefore varied dramatically in sequence and organization. Our analysis indicated that the gene amplification model of 22-kDa prolamin gene clusters can be divided into three major stages. In the first stage, rare gene duplications occurred from the ancestor gene copy accidentally. In the second stage, rounds of gene amplification occurred by unequal crossing over to form tandem gene array(s). In the third stage, gene array was further diverged by other genomic activities, such as transposon insertions, segmental rearrangements, etc. Unlike their highly conserved sequences, the amplified 22-kDa prolamin genes diverged rapidly at their expression capacities and expression levels. Such processes had no apparent correlation to age or order of amplified genes within TAG cluster, suggesting a fast evolving nature of TAGs after gene amplification. These results provided insights into the amplification and evolution of TAG families in grasses.

Among the nine species of the genus Coix, C. lacrymajobi L. and C. gigantea Koen. have 2n = 20 small chromosomes and C. aquatica Roxb. 2n = 10 large chromosomes, and are widely distributed in South and South East Asia, the first is cultivated and spread to all warmer parts of the world. The other species have restricted localized distribution. C. aquatica originated from a hypothetical diploid ancestor with 2n = 10 small chromosomes, which became longer through accumulation of duplicate segments and heterochromatin and adapted to wet habitats. C. lacrymajobi and C. gigantea originated through the same two small chromosome diploid ancestors, one of which may be common to C. aquatica, through allopolyploidy and adapted to drier conditions. All the three species can survive equally well in wet and dry localities. Aneuploid and polyploid races occurred through chromosome nondisjunction and genome doubling in C. gigantea and C. aquatica, generation of alien-addition and substitution chromosome races appears to be the further course of evolution. Six morphological varieties were differentiated in C. lacrymajobi through accumulation of gene mutations, and chromosome structural changes gave rise to a completely euchromatic chromosome race. Two aneuploid species with 2n = 32 chromosomes, both large and small, must have originated through allopolyploidy, with C. aquatica (n = 6) race as one of the parents in both, C. lacrymajobi and C. gigantea (n = 10) the second parent in Madhya Pradesh form and West Bengal form respectively. From comparison of morphology and their distribution, C. puellarum Balansa, C. ouwehandii Koord. and C. poilanei Mimeur are speculated to have originated from the three established species, through chromosomal changes and gene mutations and adapted to restricted localized areas, and C. gasteenii Simon an allopolyploid form of C. lacrymajobi and C. gigantea. A schematic diagram is presented depicting the origin, evolution and interrelationships in the genus.

Charred residues adhering to sherds of ceramic cooking vessels, pots (Fu), excavated from the Kuahuqiao site (8000–7000 cal a BP) in Zhejiang Province were examined using the method of starch grain analysis. Ancient starch grains were recovered from charred residues and were classified into 8 genera. Remains from rice (Oryza spp.), job’s-tears (Coix chinesis T.), bean (Vigna spp.), acorn (Quercus spp.) as well as possibly buckeye tree (Aesculus chinensis B.) were identified. Other starch grains from unidentified plants were also recovered. Tracheids with bordered pits from softwood fibers were also found, indicating that some food might have been processed with wooden tools. The range of starch grains in the charred residues adhering to the ceramic pots reveals the diversity of ancient plant foods at the Kuahuqiao site, Zhejiang Province.

Phytoene synthase (PSY), which is encoded by the phytoene synthase 1 (PSY1) gene, is the first rate-limiting enzyme in the plant carotenoid biosynthetic pathway. In order to examine the genetic diversity and evolution pattern of PSY1 within the Andropogoneae, sequences of 76 accessions from 5 species (maize, teosinte, tripsacum, coix, and sorghum) of the Andropogoneae were tested, along with 4 accessions of rice (Oryza sativa L.) included as outliers. Both the number and the order of exons and introns were relatively conserved across the species tested. Three domains were identified in the coding sequence, including signal peptide (SP), PSY, and highly conserved squalene synthase (SQS) domain. Although no positive selection signal was detected at an overall coding level among all species tested, the SP domain and the region upstream of the SQS–PSY domain appear to have undergone rapid evolution, as evidenced by a high d N/d S ratio (>1.0). At the nucleotide level, positive selection and balancing selection were detected only among the yellow maize germplasm and the white maize germplasm, respectively. The phylogenetic tree based on full-length sequences of PSY1-like regions supported the monophyletic theory of the Andropogoneae and the closest relationship between Zea and Tripsacum among the Andropogoneae. Coix, which was theorized to have a closer relationship with maize due to similarities in morphology and chromosome number, has been shown in this study to have diverged relatively early from the other Andropogoneae, including maize.

Graminoid molecular evolution was investigated by chloroplast genome (plastome) scale analyses. A complete plastome from Coix lacryma-jobi (Poaceae) and a draft plastome from Joinvillea plicata (Joinvilleaceae) were sequenced and analyzed. The draft plastome included conserved protein-coding loci routinely analyzed in previous studies plus one additional locus of demonstrated phylogenetic utility. The methodological approach was to directly sequence overlapping amplicons from known plastome regions. Over 100 pairs of amplification and sequencing primers were designed and positioned to flank overlapping 1,200-base pair fragments around the entire plastome. Newly determined sequences were analyzed with published plastomes from representatives of Panicoideae, Ehrhartoideae, and Pooideae. Considerable variation was found for studies within the family and even within Andropogoneae. Readily interpreted mutation patterns were observed, such as small inversions in hairpin-loop regions and indels, which were common in intergenic spacers. Maximum or near-maximum bootstrap support was observed in all analyses resolving relationships between subfamilies. However, the addition of characters from noncoding regions increased the number of parsimony-informative characters and lengthened short internal branches (Andropogoneae), better defining intergeneric relationships. Thus, characters in complete plastomes can be used over a wide scope of phylogenetic studies.

Knowledge about the composition and structure of centromeres is critical for understanding how centromeres perform their functional roles. Here, we report the sequences of one centromere-associated bacterial artificial chromosome clone from a Coix lacryma-jobi library. Two Ty3/gypsy-class retrotransposons, centromeric retrotransposon of C. lacryma-jobi (CRC) and peri-centromeric retrotransposon of C. lacryma-jobi, and a (peri)centromere-specific tandem repeat with a unit length of 153 bp were identified. The CRC is highly homologous to centromere-specific retrotransposons reported in grass species. An 80-bp DNA region in the 153-bp satellite repeat was found to be conserved to centromeric satellite repeats from maize, rice, and pearl millet. Fluorescence in situ hybridization showed that the three repetitive sequences were located in (peri-)centromeric regions of both C. lacryma-jobi and Coix aquatica. However, the 153-bp satellite repeat was only detected on 20 out of the 30 chromosomes in C. aquatica. Immunostaining with an antibody against rice CENH3 indicates that the 153-bp satellite repeat and CRC might be both the major components for functional centromeres, but not all the 153-bp satellite repeats or CRC sequences are associated with CENH3. The evolution of centromeric repeats of C. lacryma-jobi during the polyploidization was discussed.

Prolamins, the seed storage proteins of maize, sorghum and coix were also found in sugarcane. Prolamins are grouped into structurally distinct classes termed the α-, β-, γ- and δ-prolamins. Orthologues for almost all of the α-, β-, γ- and δ-prolamins classes were identified in sugarcane. In maize, there are two molecular weight classes of α-prolamins, the 22 and 19 kD α-zeins. Sugarcane also possesses both the 22 kD and the 19 kD α-prolamins, which we denote as caneins, whereas sorghum and coix contain only the 22 kD α-prolamin (α-kafirin and α-coixin, respectively). Amino acid sequence alignments of the 22 and 19 kD α-prolamins from these plants revealed that both the 19 kD α-zein and the 19 kD α-canein lack around 20 amino acids at the sixth α-helix domain. We postulate that the 19 kD α-prolamins originated from a deletion of the sixth α-helix of a 22 kD counterpart in the saccharum lineage. Saccharum and sorghum diverged around five to nine million years ago (Mya), when only the 22 kD α-prolamins existed. The 19 kD α-canein must therefore have emerged after this time. Sorghum possesses a 19 kD α-prolamin similar to that of sugarcane and maize, but it contains the sixth α-helix domain lacking in the 19 kD α-zein and the 19 kD α-canein. This sorghum α-prolamin that we called 19 kD-like α-kafirin must be the ancestor of the 19 kD α-canein. The 19 kD-like α-kafirin could also be the ancestor of the 19 kD α-zein but it is also possible that the genes encoding the 19 kD α-zein and the 19 kD α-canein have evolved separately in these close groups.

Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) has long been used as a traditional Chinese medicine for dysfunctions of the endocrine system and inflammation conditions. However, the effect of adlay seed on the endocrine system has not yet been reported. In the present study, the effects and the mechanisms of methanolic extract of adlay bran (ABM) on progesterone synthesis in rat granulosa cell were studied. ABM was further partitioned with different solvents including water, 1-butanol, ethyl acetate and n-hexane. Four subfractions named ABM-Wa (water fraction), ABM-Bu (1-butanol fraction), ABM-EA (ethyl acetate fraction) and ABM-Hex (n-hexane fraction) were obtained. ABM-Bu was further fractionated using Diaion HP-20 resin column chromatography with gradient elution. Granulosa cells were prepared from pregnant mare serum gonadotropin-primed immature female rats and challenged with different reagents including human chorionic gonadotropin (hCG 0.5 IU/ml), forskolin (10 microM), 8-bromo-adenosine-3',5'-cyclic monophosphate (8-Br-cAMP, 1 mM), A23187 (10 microM), phorbol 12-myristate 13-acetate (PMA, 0.01 microM), 25-OH-cholesterol (0.1-10 microM) and pregnenolone (0.1-10 microM) in the presence or absence of ABM-Bu (100 microg/ml). The functions of steroidogenic enzyme including protein expression of the steroidogenic acute regulatory protein (StAR) and cytochrome P450 side-chain cleavage enzyme (P450scc) protein were investigated. Expressions of both P450scc and StAR mRNA have also been explored. We found that ABM decreased progesterone production via an inhibition on (1) the cAMP-PKA and PKC signal transduction pathway, (2) P450scc and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) enzyme activity, (3) P450scc and StAR protein and mRNA expressions and (4) the phosphorylation of ERK1/2 in rat granulosa cells.

Coix is a genus in the grass family placed in the tribe Maydeae. It is closely related to maize and is also used as a crop plant. Since many valuable traits have been identified recently in Coix, it is considered to be a valuable genetic resource, particularly for maize improvement. In this study, a Coix genetic linkage map was constructed using an F2 population of 131 individuals. Eighty AFLP and 10 RFLP markers were mapped, covering a total length of 1339.5 cM with an average interval of 14.88 cM. The map consisted of 10 linkage groups, were consistent with the chromosome numbers observed cytogenetically. Both AFLP and RFLP markers were used first for genetic analysis in Coix. AFLP markers were generated by two restriction enzyme combinations, EcoRI/MseI and PstI/MseI. A total of 1349 bands were amplified, of which 140 were polymorphic. The polymorphism detection efficiency of the two enzyme combinations was compared, and utility of AFLP markers to construct the linkage map was discussed. Ten RFLP markers detected by three different probes were distributed on eight different linkage groups. The results provide a foundation to map and isolate important genes in Coix, and to investigate its genomic architecture, possible origins, and relationship with maize at the DNA level.

During the summer of 2012, some leaves of Coix lacryma-jobi (adlay or Job's tears) with leaf spot symptoms were collected from surrounding rice fields of Amol, north Iran. Four single spore isolates were obtained from diseased adlay leaves and the pathogenicity of each isolate to adlay plants was confirmed by inoculation tests based on Koch’s postulates. The pathogen was identified as Bipolaris coicis, based on both morphological characteristics and sequencing of the internal transcribed spacer region of the rDNA. This is the first report of Bipolaris coicis causing leaf blight on adlay in Iran.

Human growth hormone was expressed in transgenic tobacco seeds using the monocot tissue-specific promoter from sorghum γ-kafirin seed storage protein gene. During tobacco seed ripening, the expressed hormone was directed to the endoplasmic reticulum by a signal peptide from a Coix prolamin and was secreted into the apoplastic space, where it accounted for 0.16% of total soluble seed protein. The expressed hormone has the same amino acid sequence and receptor-binding properties as the native mature hormone.

Dihydrodipicolinate synthase (DHPS) is the main enzyme of a specific branch of the aspartate pathway leading to lysine biosynthesis in higher plants. We have cloned and characterized the DHPS-encoding DapA gene from the maize-related grass Coix lacryma-jobi. The DapA open reading frame is interrupted by two introns and encodes the 326 amino acid-long Coix DHPS protein, which is 95% identical to the maize DHPS protein. Coix DNA gel blot analysis with maize DHPS cDNA as a probe showed a single strongly hybridizing band along with faint bands. RNA gel blot analysis showed that DHPS transcripts are present in coleoptiles, embryos, endosperms, and roots but are almost undetectable in blades of young leaves of both Coix and maize. The 5′-flanking region of the DapA gene contains a TGACTC GCN4-like element located 372 bp upstream the putative translation start codon. Steady-state levels of DHPS mRNA were slightly reduced in the endosperms and embryos of the maize lysine-rich opaque2 mutants when compared with those in normal kernels. Selective binding assay with the maize Opaque2 protein (O2) showed that the GCN4-like element is not an O2 binding site, suggesting that the DHPS gene is not under the control of O2.

Polymorphism for peroxidase, esterase and acid phosphatase Isozymes in pollen grains of major Indian maize land races known as ‘Sikkim primitives’ were studied and compared with primitive races of maize of Mexican origin and five species of teosinte, Coix lacryma-jobi, Chionachne koenigii and Sorghum bicolor. The isozyme patterns and the resulting dendrograms revealed similarities in primitive Indian maize landraces from Northeastern Himalayan region and Nal-Tel an ancient indigenous race of Mexico. Further, the Asiatic taxa of Maydeae, C. lacryma-jobi and Ch. koenigii differed widely from each other and also from the Zea spp. Presence of greater diversity among the Indian maize collections was also evident from the present analysis.

The seed storage proteins of Coix, sorghum and maize are codified by homologous genes which are coordinately expressed in the endosperm in a temporal-specific fashion. Opaque2 (O2), a bZIP protein originally isolated from maize, has been described as a transcription activator of α- and β-prolamin genes. The isolation and characterization of cDNA and genomic clones encoding the Opaque2 homologue from Coix are reported here. The coding region of the Coix O2 gene is interrupted by five introns and codifies a polypeptide of 408 amino acids. Comparison of the deduced amino acid sequence with two different sequences of maize O2 protein showed that the Coix O2 protein is similar to the maize O2 isolated from W22 maize inbred line. The Coix O2 protein has the same binding specificity and expression pattern of the maize O2. The O2 proteins together with OHP1, OsBZIPPA, SPA, CPRF2 and RITA1 were assigned to one of the five bZIP plant families in an updated classification of plant bZIP according to bZIP domain similarity.

Six populations of Coix lacryma-jobi with distinct characters were studied to evaluate the presence of autonomous apomixis and pseudogamy, and to determine its implications and effects in the calculations of the rate of breeding. For this reason, two types of genetic markers were used-morphological markers and isozymatic markers.

Among a natural population of Coix gigantea a pentasomic (2n+3; 2n=23) and a hexasomic (2n+4; 2n=24) were isolated and studied cytologically for the first time. The extra homologous chromosomes in the pentasomic plant did not disturb the behaviour of the rest of the bivalents through meiosis except for the rare occurrence of a few univalents. The majority of PMCs at diakinesis showed nine bivalents and a pentavalent. The various anaphase segregations led to the formation of sub- and super-haploid male gametes with from n−1 (n=9) to n+4 (n=14) chromosomal constitutions. The four extra homologous chromosomes in the hexasomic plant, however, disturbed meiosis leading to sticky chromosomal configurations and sterility. At diakinesis, the majority of the PMCs showed nine bivalents and a hexavalent.

A series of chromosomal variants has been isolated from an open-pollinated progeny of interspecific hybrids between aneuploids of Coix gigantea (2n=18–24) and Coix aquatica (2n=10). The interspecific hybrids (2n=14, 15 and 16) produced several types of gametes not only with different chromosome numbers but also comprised of varied permutations and combinations of gigantea and aquatica chromosomes. This was evident when the open-pollinated progeny obtained from these hybrids was screened chromosomally. Two such open-pollinated experimental progenies were studied in two successive years (1983 and 1984) and plants with from 2n=10 to 2n=21 chromosomes were isolated. Chromosomal configurations at diakinesis in all the variants revealed frequent pairing between the gigantea and aquatica chromosomes. This indicated that the two species are phylogenetically closely related. Restoration of ‘pure’ parental species from the F1 hybids and chromosomal variants through genomic segregation and spontaneous back-cross are unique and noteworthy features.

Five cloned histone H3 and H4 genes from maize have specific 5′ non-transcribed regions. Blot hybridization of each 5′ region to DNA from different maize inbred lines showed that the H3 and H4 multigenic families are organized into subfamilies. Each subfamily has a specific environment and contains a different (from 4–16) number of gene copies. H3 and H4 subfamilies with similar environments as those found in maize were shown to exist in the genomes of more or less related plants, including perennial teosinte, sorgho, sugar cane and Coix. Such observations may contribute to establishing phylogenetic relationships at a molecular level between different plants and thus highlight some of the evolutionary mechanisms of the genomes of higher plants.

Fronds of a fern of theAsplenium acrobryum complex were traditionally used as a source of salt in the inland areas of Papua New Guinea. All previously published reports of the use of fern salt in the botanical and ethnobotanical literature had erroneously identified the species involved asAsplenium nidus, one of the common bird’s-nest ferns. A chemical analysis of the salts contained in the ash of the salt fern as well as those species related toAsplenium nidus revealed that the bulk of the inorganic component was potassium, calcium and chloride. These results are comparable to those published for the salt-yielding grass,Coix gigantean. There is no obvious chemical reason why onlyAsplenium acrobryum should be used for salt production in preference toAsplenium nidus and related species. In view of the low levels of consumption of such salt it is unlikely that there will be any cases of potassium toxicity attributable to the use of these plants.

Job’s-tears—a cultigen of great antiquity—is grown in South Asia mainly by the natives of various ethnic groups of Mongolian origin. The plant is used as a minor cereal and fodder. The northeastern region of India is a centre of variability for the genus Coix. It is considered that this plant was introduced here either by the pestoral Aryan invaders, who grew it on the slopes of the Himalayas or during Mongolian conquests when the crop got distributed from the eastern Himalayas to lower subtropical terrains. During plant explorations conducted since 1970 in the northeastern region of India, much variability has been recorded for this crop. The various soft-shelled races now grown in this tract by the tribes are the result of conscious folk domestication and must have been selected for easy hulling and good kernel type. Based on this, and on information on domestication, cultivation and economic usages of this plant among the natives, some findings are presented in this paper.

Callus was obtained from segments of immature inflorescence of Coix lacryma-jobi cultured on N6 medium containing 1–2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 3–5% sucrose. Plantlets were regenerated when embryogenic calluses were transferred onto MS medium with 0.5 mg/l kinetin and 0.01 mg/l naphthaleneacetic acid (NAA). Regenerated plants had the diploid chromosome number (2n=20).

Meiotic behaviour of a monosomic-trisomic (2n−1+1=20) of Coix gigantea (Poaceae) showed that, although there are a few segregational irregularities, it is essentially mostly normal. The plant is healthy and semifertile, and through the formation of viable male and female gametes of variable chromosomal constitution it is expected to produce simple and complex aneuploids in its progeny. The origin is believed to have been through nondisjunction of two bivalents, a large and a small in a diploid.

Antibody systems were produced in rabbits by immunization with antigen systems from seed ofZea mays, Z. mexicana andTripsacum dactyloides. Antigen systems were generally ranked in the following order using Zeaantibody systems: Zea > Tripsacum > Elyonurus ≈ Bothriochloa > Coix ≈ Manisuris > Andro-pogon ≫ Triticum. Tripsacum antibody systems also ranked these antigen systems in a similar order except that Tripsacum > Zea, and Manisuris ≈ Bothriochloa ≈ Elyonurus. Some Tripsacum species produced anomalous results. Serology suggests that Zea and Tripsacum should be placed together in the subtribe Tripsacinae of the tribe Andropogoneae: the tribe Maydeae is probably an unnatural assemblage. Tripsacum and Zea have probably evolved from ancestors with affinities to the subtribes Rottboellinae and Bothriochloeae of the tribe Andropogoneae. The high degree of serological correspondence shown by Elyonurus to Zea suggests a close common ancestry, but Manisuris appears no more similar to Tripsacum than do other genera of the Rottboellinae and Bothriochloeae. By polyacrylamide gel electrophoresis, no differences were found between maize and teosinte from Mexico and north Guatemala. Teosinte from south Guatemala consistently lacked bands present in both maize and Mexican teosinte but shared no greater similarity to Tripsacum and cannot therefore be considered as tripsacoid. The high degree of band homology between maize and Mexican teosinte supports a parental relationship and it is suggested that Mexican teosinte represents the germ plasm from which maize was domesticated. Neither electro-phoretic nor serological results supported the hypothesized hybrid (viz. Zea x Manisuris) origin of Tripsacum.

Desynapsis leading to total sterility was observed in one plant of Job's tears, Coix lacrymajobi. L., during summer months, while the other neighbouring plants were quite normal. Pachytene chromosome pairing in the desynaptic was apparently normal but a diakinesis and metaphase I several univalents were found. Perennial suckers of this plant in the rainy season showed regular meiosis and good fertility. It is probable that desynapsis was caused by an interaction of genotype and environment, in which the desynaptic gene became activated by the rapid increase in summer temperatures whilst its effect on chromosome synapsis was completely lost in the cooler periods.

Two cases of reversion to diploidy were observed in autotetraploid Job's tears (Coix lacryma-jobi L. 4n = 40) out of a total of 1,112 plants examined over a period of 7 years. One of these was a trisomie (2n = 21) and the other a disomic (2n = 20), derived from apomictic development ofn+1 andn maternal gametes of the tetraploid, respectively. In some respects both these derivatives differed from the original diploid that gave rise to the tetraploid through colchicine treatment. The potentialities of such reversions in the evolution of new diploid races are discussed.