Germinating seeds of the mung bean, <i>Vigna radiata</i> (Fabaceae), as a model for the preliminary evaluation of cytotoxic effects of drugs

Cytotoxic properties of plant extracts and drugs being developed for cancer treatment are usually evaluated by a variety of in vivo and in vitro tests carried out in animal or plant based models. In the present study we have evaluated the possibility of using the germinating mung beans (Vigna radiata), for rapid and inexpensive screening of drugs exhibiting cytotoxic properties. Mung beans were allowed to germinate either in tap water or in different drug solutions, and parameters like percent germination, increase in radicle length, change in seedling weight and mitotic index of apical root meristems were determined at two time intervals coinciding with the time at which the radicle length in control group was 1.0 to 1.5 cm (time 0, T0) and 48 h later (T48). Methanol extract of Calotropis procera latex as well as drugs like podophyllotoxin, cyclophosphamide, cyproheptadine and aspirin produced a dose-dependent inhibitory effect on seed germination, seed weight gain, radicle growth and mitotic index in the radicle meristems. The inhibitory effect of some of the drugs tested was associated with reduction in water imbibition. Some of the drugs at higher concentrations allowed seed germination to take place but produced radicle decay and seedling weight loss. Our study shows that germinating V. radiata beans could be used as a convenient model for the preliminary screening of drugs exhibiting cytotoxic properties.     

Storage lipids and proteins of <i>Euterpe edulis</i> seeds

Comparative studies on fatty acid and protein composition of the endosperm and embryo of palmito (Euterpe edulis Martius) were conducted using gas-liquid chromatography and sodium dodecyl sulfate–polyacrylamide gel electrophoresis. On a dry weight basis, the embryo contained extremely lower amounts of lipids and proteins than did the endosperm, which was associated with the scarce lipid and protein bodies previously reported in axis and cotyledon. The fatty acid composition also exhibited differences between both tissues: (I) the fatty acid diversity was greater in embryo than in endosperm; (II) embryo and endosperm contained predominantly linoleic, palmitic, oleic and stearic acids even though the relative values were different for each tissue. As compared to other palm species, the higher fatty acid unsaturation in Euterpe edulis seed could be involved in the previously reported short longevity and recalcitrant behavior during storage. Proteins of both tissues were heterogeneous in molecular mass. Some proteins were tissue-specific, but other were common, among them a highly glycosylated protein which migrated at about 55 kDa. We hypothesize that the latter, also reported in all previously studied palm species, is one of the proteins characterizing the Arecaceae family.     

<i>Claroideoglomus etunicatum</i> improved the growth and saline– alkaline tolerance of <i>Potentilla anserina</i> by altering physiological and biochemical properties

To investigate the effects of arbuscular mycorrhizal (AM) fungi on the growth and saline–alkaline tolerance of Potentilla anserina L., the seedlings were inoculated with Claroideoglomus etunicatum (W.N. Becker & Gerd.) C. Walker & A. Schüßler in pot cultivation. After 90 days of culture, saline–alkaline stress was induced with NaCl and NaHCO₃ solution according to the main salt components in saline–alkaline soils. Based on the physiological response of P. anserina to the stress in the preliminary experiment, the solution concentrations of 0 mmol/L, 75 mmol/L, 150 mmol/L, 225 mmol/L and 300 mmol/L were treated with stress for 10 days, respectively. The mycorrhizal colonization rate, mycorrhizal dependence, chlorophyll content, malondialdehyde content, antioxidant enzyme activities, osmoregulation substances content and water status were measured. The results showed that with the increase of NaCl and NaHCO₃ stress concentration, mycorrhizal colonization rate, colonization intensity, arbuscular abundance and vesicle abundance decreased, and reached the lowest value at 300 mmol/L. Strong mycorrhizal dependence was observed after the symbiosis with AM fungus, and the dependence was higher under NaHCO₃ treatment. Under NaCl and NaHCO₃ stress, inoculation with AM fungus could increase chlorophyll content, decrease malondialdehyde content, increase activities of superoxide dismutase, peroxidase and catalase, increase contents of proline, soluble sugar and soluble protein, increase tissue relative water content and decrease water saturation deficit. It was concluded that salt–alkali stress inhibited the colonization of AM fungus, but the mycorrhiza still played a positive role in maintaining the normal growth of plants under salt–alkali stress.     

谷物干燥机烘干温度对种子发芽率的影响分析

针对谷物干燥机烘干后的种子存在着发芽率降低的问题,本研究对影响种子发芽率的诸多因素进行了归纳,对温度参数进行检测,试验前后的种子发芽率,温度参数对烘干种子发芽率的影响进行了研究,提出了一种谷物干燥机烘干温度分析方法,利用合理布置的比对温度传感器,对谷物干燥机内的实时温度进行了监测。研究结果表明,烘干后种子发芽率降低的主要原因是谷物干燥机的机配温度传感器存在误差,影响烘干温度的准确性;谷物干燥机热风道结构的设计,温度最高位置会导致相邻谷物温度超差;谷物干燥机内温度分布不均匀导致局部谷物温度超差及机配温度传感器在谷物干燥机内的布置不合理,未反应机内最高温度。     

Cloning and characterization of 66 kDa streptavidin-binding peptides (SBP) of <i>Pisum sativum</i> L. embryo specific to var. Alaska

The aim of the current research was to clone and to characterize the partial 66 kDa streptavidin-binding peptide (SBP) found in the germinated embryos of Pisum sativum L. var. Alaska. The pea (P. sativum var. Alaska) embryos possess prominent 66 kDa SBPs that gradually disappeared after few hours of germination in germinated embryos, but not in the cotyledons. The total RNA was isolated from embryos of P. sativum but could not be isolated from the cotyledons. The partial nucleotides sequences of 66 kDa SBPs of embryonic stalk (P. sativum var. Alaska) were cloned and identified using pMOSBlue vector. 66 kDa (SBP) gene from the embryos of P. sativum var. Alaska possesses 327 bp having an open reading frame (ORF) region in a part of the gene that encoded for 108 amino acids. Alignment showed similarity among 66 kDa SBPs P. sativum var. Alaska, with P. sativum seed biotinylated protein (SBP65) and P. sativum sbp65a mRNA with DNA distance matrix between 0.0094 to 1.2676. MALDI-TOF mass spectrometry analysis of 66 kDa (SBP) proteins showed it had similar short peptides to 19 proteins found in different organisms, especially Convicilin precursor, and the seed biotinylated protein in P. sativum. The alignment results of both nucleotide sequences and amino acid residues either from cloning or MALDI-TOF-MS showed differences with related species, especially P. sativum. No mRNA was found in the cotyledons during seeds germination, which means no metabolic activities and this part may act only as food reservoirs for growing newly embryos.     

基于分段烘干工艺的玉米种子多用烘干系统设计

为提高玉米果穗加工性能及烘干设备的利用率,减少能源浪费,设计了一套采用分段烘干工艺的多用烘干系统.根据玉米种子水分的不同,采用不同的热风循环方式,使同批次的干、湿仓果穗的含水量基本一致.对烘干机理进行了分析研究,进行了烘干系统主要部件的设计.试验表明,分段烘干工艺比一次烘干工艺降低能耗及生产成本20%～30%,平均降水...     

The chloroplast genome comparative characteristic of artificial breeding tree,a case about <i>Broussonetia kazinoki</i> × <i>Broussonetia papyrifera</i>

Broussonetia kazinoki × Broussonetia papyrifera (ZJGS) is a hybrid species in Moraceae family, which has a very complicated hybrid origin. The excellent characteristics of fast growth, strong soil and water conservation ability, high leaf protein content and stem fiber content in ZJGS make it both ecological benefits in the mining area and economically valuable. This study aims to further understand ZJGS and other Moraceae taxa through the ZJGS chloroplast (cp) genome structure and the comparison with 12 closely related Moraceae species. Among the 13 Moraceae species, the cp genome length of seven Broussonetia species (ranges from 160,239 bp to 162,594 bp) is larger than that of six Morus species (ranges from 158,459 bp to 159,265 bp). Among the 77 shared protein-coding genes (PCGs) in Moraceae species, the obvious positive selection of Ka/Ks ratios acted on petD and rpl16 genes of B. kazinoki and B. papyrifera, respectively. Phylogenetic analysis based on shared PCGs from 28 species shows that ZJGS is closely related to maternal B. kazinoki. These findings provide data support for the origin of ZJGS hybridization and provide genomic resources for future ZJGS resource development and molecular breeding.     

Expression analysis of <i>OsSERK</i>, <i>OsLEC1</i> and <i>OsWOX4</i> genes in rice (<i>Oryza</i> sativa L.) callus during somatic embryo development

Somatic embryogenesis is an asexual reproduction process that occurs in many plant species, including rice. This process contains several totipotency markers such as Somatic Embryogenesis Receptor-like Kinase (SERK), Leafy Cotyledon1 (LEC1) and WUSCHEL-Related Homeobox4 (WOX4) and also a helpful model for embryo development and clones and transformations. Here, we report the gene expression during somatic embryo development correlates with regeneration frequency in 14 Javanica rice (pigmented and non-pigmented) using modifified N6 media supplemented with Kinetin (2.0 mg/L) and NAA (1.0 mg/L). Although there have been advances in understanding the genetic basis of somatic embryogenesis in other varieties, rice is still unexplored, especially during somatic embryo development. Moreover, for the formation of callus induction from immature embryos, 2,4-D (2.0 mg/L, 3.0 mg/L) was used. This study analysed the gene expression of OsSERK, OsWOX4 and OsLEC1 genes through RT-PCR analysis. Higher expression of the OsLEC1 gene indicates that their function may correlate in the in vitro with the high response of rice after transfer to regeneration media. This study found that rice varieties of pigmented rice (MS Pendek and Gogoniti II) and non-pigmented rice (Pandan Ungu) showed high regeneration frequency, showing higher OsLEC1 expression than other varieties because OsLEC1 promotes the maturation of somatic embryos in plant regeneration on day 14. However, the contrast with Genjah nganjuk may be effective because of other regulatory genes. RT-PCR analysis showed OsSERK had less expression level than OsLEC1 and OsWOX4 in the varieties, which correlate with the percentage of plant regeneration, but not for Gogoniti II. In conclusion, the higher percentage of plant regeneration correlates with the higher expression level of OsLEC1 at day 14 of media regeneration of rice.     

Water loss at normal enamel histological points during air drying at room temperature

This in vitro study aimed to quantify water loss at histological points in ground sections of normal enamel during air drying at room temperature (25°C) and relative humidity of 50%. From each of 10 ground sections of erupted permanent human normal enamel, three histological points (n = 30) located at 100, 300 and 500 μm from enamel surface and along a transversal following prisms paths were characterized regarding the mineral, organic and water volumes. Water loss during air drying was from 0 to 48 h. Drying occurred with both falling and constant‐drying rates, and drying stabilization times (T_eq) ranged from 0.5 to 11 h with a mean 0.26 (±0.12)% weight loss. In some samples (n = 5; 15 points), T_eq increased as a function of the distance from the enamel surface, and drying occurred at an apparent diffusion rate of 3.47 × 10^?8 cm² s^?1. Our data provide evidence of air drying resulting in air replacing enamel's loosely bound water in prisms sheaths following a unidirectional water diffusion rate of 3.47 × 10^?8 cm² s^?1 (from the original enamel surface inward), not necessarily resulting in water evaporating directly into air, with important implications for transport processes and optical and mechanical properties.     

<i>Trypanosoma rangeli:</i> growth in mammalian cells <i>in vitro</i> and action of a repositioned drug (17-AAG) and a natural extract (<i>Artemisia sp</i>. essential oil)

Trypanosoma rangeli and T. cruzi are both parasitic unicellular species that infect humans. Unlike T. cruzi, the causative agent of Chagas disease, T. rangeli is an infective and non-pathogenic parasite for humans, but pathogenic for vectors from the Rhodnius genus. Because both species can coexist in different hosts and overlap their infective cycles but very little is known about the infection of T. rangeli in mammalian cells, we decided to characterize both the development of this parasite in cell culture and the effect of therapeutic agents with potential trypanocidal action on it. We found that T. rangeli exhibits a cycle of infection in Vero cells similar to that for T. cruzi and that the repurposed drug, 17-AAG, and the natural extract Artemisia sp. essential oil produce a toxic effect on epimastigotes showing a trypanocidal action from the fifth day of culture. Both treatments also affected the infection of trypomastigotes and reduced the capacity of replication of amastigotes of T. rangeli. Since T. cruzi / T. rangeli coinfection cases have been reported, the finding of drugs with potential activity against both species could be significant in the future. Furthermore, studies of susceptibility of both species to drugs could also help to know the different mechanisms of pathogenicity in humans displayed by T. cruzi that are absent in T. rangeli     

Brief Note : Improved <i>in vitro</i> embryo development of stenospermic grape by putrescine

The goal of this study was to determine the effect of putrescine, added to the culture medium, on the in vitro development of stenospermic grape (Vitis vinifera L) embryos. The cross breedings of Perlón x G.C88552 and Perlón x Argentina were used. 0 (control), 2 and 4 mM of putrescine were added to the immature seed’s culture medium. In Perlón x Argentina, 2mM of putrescine statistically increased the percentage of total embryos, direct germination, polyembryos and normal plants. In Perlón x G.C88552, only 2 mM of putrescine increased all the variables considered, eventually tripling the percentage of normal plants obtained. The results suggest that the endogenous concentration of putrescine may be a growth limiting factor. Adding putrescine to the culture medium of immature grape seeds is a legitimate resource to significantly increase the results of this technique.     

Genome-wide identification of <i>U-box</i> gene family and expression analysis under abiotic stresses in <i>Salvia miltiorrhiza</i>

Plant U-box (PUB) E3 ubiquitin ligases play important roles in hormone signaling pathways and in response to different abiotic stresses, but little is known about U-box genes in Danshen (root of Salvia miltiorrhiza Bunge). Here, we identified and characterized 70 SmPUB genes based on its genome sequence. Phylogenetic analysis of U-box genes from S. miltiorrhiza and Arabidopsis suggested that they can be clustered into seven subgroups (I–VII). Typical U-box domains were found in all identified SmPUB genes through the analysis of conserved motifs. Moreover, qRT-PCR was applied to analyze the relative expression levels of U-box genes in S. miltiorrhiza roots and leaves under PEG-induced water deficit and salt stresses. Results revealed that the SmPUB genes exhibited stronger response to drought than to salt stress. To the best of our knowledge, this report is the first to perform genome-wide identification and analysis of the U-box gene family in S. miltiorrhiza, and the results provide valuable information for better understanding of the function of U-box in S. miltiorrhiza.     

Antiproliferative effect of extracts of <i>Sida rhombifolia</i> L. (Malvaceae) on the <i>Allium cepa</i> cell cycle

Field collected roots of four populations of Sida rhombifolia were used for preparing aqueous decoctions at two concentrations: 4g/L; and 16g/L. Afterwards, we used three groups of six onion (Allium cepa) bulbs for testing each population. Slides were made with all bulbs through the smashing technique. Cells in all phases of the cell cycle of A. cepa were analyzed. The mitotic index (% of cells in mitosis) was calculated, and the statistical analysis through the χ2 test was carried out at 5% probability. The results showed that the aqueous extracts of S. rhombifolia have antiproliferative activity at high concentrations. Practically no chromosomal aberrations were induced by treatments.     

<i>Agrobacterium rhizogenes</i> vs auxinic induction for <i>in vitro</i> rhizogenesis of <i>Prosopis chilensis</i> and <i>Nothofagus alpina</i>

The induction and improvement of in vitro rhizogenesis of microshoots of Prosopis chilensis (Mol.) Stuntz and Nothofagus alpina (Poep. et Endl. Oerst.) were compared using Agrobacterium rhizogenes (Ar) versus indole-3-butyric acid (IBA) in the culture media. Microshoots of P. chilensis (1-2 cm length), coming from in vitro grown seedlings, were cultivated in a modified Broadleaved Tree Medium (BTMm) containing half salt concentration of macronutrients and 0.05 mg.L^-1 benzilaminopurine (BAP). After 30 days, microshoots with 2-4 leaves were selected and cultured in BTMm-agar in presence or abscense of Ar and in combination with IBA. For N. alpina, the apical shoots with the first 2 true leaves, from 5 weeks old seedlings, were cultured in the abovementioned medium, but with 0.15 mg.L^-1 of BAP. After 2 months, microshoots with 2-3 leaves were selected and cultured in BTMm-agar, supplemented with 5 mg.L^-1 IBA or in liquid BTMm on perlite and, in the presence or absence of A. rhizogenes (Ar) and in combination with 3 mg.L^-1 IBA. Rooting in P. chilensis reached 100.0% when Ar infection was produced in the presence of IBA, increasing both, the number and dry weight of roots. In N. alpina, 90.0% of rooting efficiency was obtained when Ar infection was produced in liquid culture and in the absence of auxin.     

Isolation and molecular characterization of a <i>cax</i> gene from <i>Capsella bursa-pastoris</i>

A new cation exchangers (CAXs) gene was cloned and characterized from Capsella bursapastoris by rapid amplification of cDNA ends (RACE). The full-length cDNA sequence of cax from C. bursa-pastoris (designated as Cbcax51) was 1754 bp containing a 1398 bp open reading frame encoding a polypeptide of 466 amino-acid residues with a calculated molecular mass of 50.5 kDa and an isoelectric point of 5.69. The predicted CbCAX51 contained an IMP dehydrogenase/GMP reductase domain, two Na⁺/Ca²⁺ exchanger protein domains. Comparative and bioinformatics analyses revealed that CbCAX51 showed extensive homology with CAX from other plant species. The expression analysis by different treatments indicated that Cbcax51 could be activated by cold triggering and was related to the cold acclimation process, but its expression is regulated negatively by drought and not affected by ABA or salt.     

Differential metabolome landscape of <i>Kadsura coccinea</i> fruit tissues and potential valorization of the peel and seed tissues

Kadsura coccinea (Lem.) is a woody wine plant with a peculiar fruit enriched in important health-promoting compounds. The non-editable part of the fruit, i.e., the seed and peel, represents more than 60% of the fruit and is considered a biowaste. This significantly restricts the development of the K. coccinea fruit industry. Clarifying the metabolic components of the different fruit parts can help to improve the utilization rate and valorization of K. coccinea. Herein, we evaluated K. coccinea fruit peel, pulp, and seed using widely-targeted metabolomics and quantified a set of 736 bioactive compounds from 11 major metabolite classes. The most prominent metabolite classes included lipids, amino acids, flavonoids, and lignans. Furthermore, our results emphasized a significant accumulation of flavonoids in pulp tissues, while alkaloids and lignans were abundant in peel and seed tissues, respectively. A total of 183 metabolites were differentially accumulated among the three tissues. Procyanidin C2, rutinoside, 2-hydroxyoleanolic acid, 5-hydroxymethyluracil, nootkatol, isoquercitrin, isohyperoside, quercetin-7-O-glucoside, hyperin, and rutin showed elevated accumulation in the peel. In the seed, kadsuralignan G, kadcoccilactone A, kadsuralignan H, lysoPE 20:5, iso-schisandrin ethyl alcohol, and kadangustin were significantly enriched. Our results highlight the diverse metabolome composition of K. coccinea fruit parts, which can be further exploited for its valorization in various industries.