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In this work we used semi-isolated heart of the cockroach Nauphoeta cinerea for the investigation of the pharmacological effects of extracts (aqueous, 1:1, 1:2, 1:4 and 1:8) from Nephrolepis exaltata L. leaves, a popular ornamental fern considered to be safe. The use of insects in experimental studies has grown due to the easy handling, proliferation/growing assuring its rapid obtention, and absence of ethical issues. An aqueous extract 0.2 % was obtained after maceration of 1 g N. exaltata leaves powder with 20 mL of distilled water (1:20). Diluted extracts in water were obtained to have the following proportion 1:1, 1:2, 1:4 and 1:8. Experiments (n=4) consisted of 200 µL addition onto semi-isolated heart preparation of N. cinerea with concomitant heart beating counting. Aqueous, 1:1 and 1:2 extracts paralyzed completely the heart beatings of cockroachs (p<0.05 compared to saline control), but not 1:4 or 1:8, which showed only a slight decline (p>0.05 compared to saline control). A preliminary thin layer chromatography showed the presence of unidentified terpenoid in aqueous extract of N. exaltata. These pharmacological findings of N. exaltata can be exploited for future use as insecticide or as dose-dependently cholinergic agent.

Diseases are a major cause of post-harvest losses depending on season, region and management practices. Chemical control is the most used but with serious consequences for human health and the environment. This forces us to carry out more exhaustive studies on botanical products. The general objective of the present study was to evaluate the effect of citrus extracts for the control of pathogens that cause post-harvest diseases in tomato fruit. The product to be evaluated is of botanical origin from citrus extracts. Doses were evaluated (0, 666, 1000, 2000, 4000, 8000 ppm). The treatments were located at a temperature of 25°C±2 and 45% relative humidity (rH). The design used corresponded to a completely random design. The least significant difference was estimated by Tukey Multiple Range test at P=0.05. The statistical tests were performed through the SAS computer program. The results indicate that the pathogens detected and identified correspond to Alternaria tenuissima; Botrytis cinerea; Cladosporium fulvum; Colletotrichum coccodes; Fusarium oxysporum; Geotrichum candidum; Rhizopus stolonifer and Stemphylium macrosporoideum. Our conclusion is that the efficient doses correspond to 666, 2000 and 8000 ppm. With the application of citrus extracts, the damage percentage of tomato fruit was reduced in relation to the control treatments. Based on the results with the application of citrus extracts, the shelf life of the tomato was lengthened.

Phytosanitary inspectors play an important role in diagnosing diseases in foreign plant material. However, some deficiencies have been detected in the detectionc ausing the entrance of many microorganisms. Therefore, it was of great interest to detect the presence of Clavibacter michiganensis subsp. michiganensis (Cmm) in foreign tomato and chili seed in the agricultural area of Sinaloa, Mexico, besides the growth and cell density of Cmm was evaluated in different selective media under continuous illumination and photoperiod. The results indicate that seeed of 35 varieties of tomatoes was collected; while for Chili seed were 18. This study was supported by farmers (225) which represent 79% of all growers and 32 business engaged in the sale of agro-supplies, provided seeds of varieties and hybrids. Those growers are from six areas (Culiacan, El Tamarindo, Navolato, Culiacan, El dorado and Badiraguato). For detection of Cmm in tomato seed, from 35, only four was variability considering Immunochromatography and ELISA techniques; however, considering chemical and physiological test, the result was negative. Similar results were in 18 varietes of chili seed, where eight showed variability to detect Cmm, and negative by chemical and physiological test. According to the growth and cell density of Cmm, the optimal medium was YDC under pH stable and continuous light conditions. It is recommended to consider the fusion of diagnostic techniques in the emission of a result.

The halophyte Distichlis palmeri (Vasey) is a plant resource with high potential to be harvested in the coastal areas of northwestern Mexico; enlarge the knowledge and domestication for its incursion into the agricultural sector, plays an important role for arid areas with saline intrusion problems. However, its productivity depends on the supplementary supply of nitrogen, as well as other essential macro and micronutrients. The microorganisms considered beneficial are an alternative to chemical fertilization, highlighting those Plant Growth Promoting Bacteria (PGPB). In the present study, the inoculation of the Bacillus amyloliquefaciens (B.a.) as a halobacterium PGPB was evaluated to know the response in seeds of Distichlis spicatai obtained from natural population from colorado river in Delta north of the Gulf of California. Wild seed was collected and germinated previously inoculated with B. a., and sowed in germinated beds. Later, seedlings were planted under field and salinity conditions in the coast of Hermosillo, Sonora. Three treatments were examined (T1: B.a., T2: Chemical fertilization, T3: Negative control), with four repetitions each treatment. Each repetition consisted of experimental plots of 5 x 5 m, with a separation of 1 m between them. The harvest was carried out 600 days after sowing. The results indicate that treatments inoculated with halobacteria B.a., showed significant results in crude protein, non-protein nitrogen, neutral detergent fiber and acid detergent fiber, as well as spike length and number of seeds. The results obtained suggests the feasibility of biofertilizers where biomass and seed production are significant compared to non-inoculated controls.

Aflatoxins, produced by Aspergillus spp., are strongly toxic and carcinogenic fungal secondary metabolites. Aflatoxin biosynthesis is a complex process and involves at least 30 genes clustered within an approximately 75 kB gene cluster. In this paper, we reviewed current status of the researches on the characterized structural genes involved in aflatoxin biosynthesis and their roles in aflatoxin-producing fungi, especially in A. flavus and A. parasiticus, which will improve our understanding of the mechanism of aflatoxin biosynthesis and regulation and provide reference for further study.

This study purposed to study the Preservative agents that are required to ensure that manufactured foods remain safe and unspoiled; work was conducted to evaluate the efficacy of essential oils from two eucalyptus species, Eucalyptus grandis and Eucalyptus crebra in food preservatives; to run this experiment flesh eucalyptus leaves collected from Ruhande Arboretum forest were submitted to hydrodistillation and yields(amount) of 0.38 and 0.34 % for Eucalyptus grandis and Eucalyptus crebra were obtained, respectively. Phaseolus vulgaris, Sorghum condatum, cooked Ipomoea batatas (sweet potatoes) and bread were the sample foods used to assess their preservative efficacy. Acanthoscelides obtectus and Stophilus oryzae were used as pests for Phaseolus vulgaris and Sorghum condatum respectively. For bread and cooked Ipomoea batatas, Rhizopus nigricans are used to assess the efficacy of these two essential oils to inhibit their growth; the obtained results revealed that those essential oils could act as insecticide in the storage of Phaseolus vulgaris and Sorghum condatum. Essential oil from Eucalyptus grandis protected these two foods against pests in the periods of 4 and 9 days, respectively while essential oil from Eucalyptus crebra protected them for the periods of 6 and 11days, respectively.

Many agriculturally important properties such as heterosis, inbreeding depression, phenotypic plasticity, and resistance for biotic and abiotic stresses are thought to be affected with epigenetic components. New discoveries related with epigenetics are likely to have a major impact on strategies for crop improvement in rice breeding. However, assessing the contribution of epigenetics to heritable variation in plant species still poses major challenges. Methylation of cytosine in DNA is one of the most important epigenetic mechanisms in plants. DNA methylation not only plays significant roles in the regulation of gene activity, but also it is related with genomic integrity. Although most of next generation sequencing (NGS) technologies do not require the use of target specific primer pairs to identify and study DNA cytosine methylation, validation studies of NGS uses selective primer pairs. Bisulfite sequencing technique is a gold method for DNA cytosine methylation studies. However, bisulfite sequencing requires the development of bisulfite primer pairs to selectively study DNA sequences of interest. In this study 9 bisulfite specific primer pairs were identified and validated. These primer pairs successfully amplified bisulfite converted and unconverted genomic DNA extracted from radicle and plumule of rice (Oryza sativa L.) seedlings. Results of the present study clearly revealed the occurrence of CG, CHG and CHH (H stands for C. T or A nucleotides) contents in studied DNA sequence targets were different indicating potential role of DNA cytosine methylation in these genes. Primer pairs reported in this study could be used to detect DNA methylation which is one of the most important epigenetic mechanisms affecting the development, differentiation or the response to biotic and abiotic stress in rice (Oryza sativa L.).

The age-old battle between plants and viruses has many twists and turns. Plants acquired the RNAi factors to checkmate the viruses and the viruses encode VSRs to defeat RNAi for their own survival. Plants designed mechanisms to neutralize the toxic effects of VSRs and the viruses, in their turn, use host microRNAs to strengthen their infection processes. The infightings between these two entities will take different shapes with prolonged evolution and accordingly the researchers will dig these novel forms of duels not only to throw lights in the involved mechanisms but also to manipulate various antiviral strategies. Some of the research courses that might come up in the immediate future are discussed.

Ilex paraguariensis, also known as ‘Yerba mate’, occurs naturally in Argentina, Brazil and Paraguay and is also grown in these countries with different intensities. Leaves and branches of this plant are used in the preparation of a stimulant beverage that beside social importance has notorious health impact. However, the cultivated herbs present low productivity, due to deficiencies in cultivation and harvesting techniques, as well as due to the abiotic stresses that this species is subject to. The discovery and characterization of cold response mechanisms in plants such as Arabidopsis thaliana, began research in order to unravel the physiological and molecular mechanisms in response to cold in other plant species. In this work, we studied the physiological response observed in Ilex paraguariensis plants submitted to low temperatures (0°C), with or without a pre-moderate acclimatization treatment period of (8°C). Our results suggest the existence of an acclimation response in Ilex paraguariensis, similar to that described in other species of the same temperature.

The use of novel PGPR as bio inoculant is an alternative sustainable agricultural practice to improve soil health, grain quality, increase crop productivity, and conserve biodiversity. The aim of this study is to isolate, and characterized PGP bacteria colonizing tef rhizosphere during the seedling stage. For this concern, 426 samples of tef (Eragrostis tef) rhizosphere soils and roots were collected from East Shewa zone, Oromia regional state. 200 morphologically different bacterial pure colonies were isolated and screened for their PGP traits and biocontrol properties. Among these 40.5% isolates were positive for phosphate solubilization. 36% were positive for IAA production, 4.5% were positive for ammonia production, 19 % were positive for (EXPS), 15.5% were positive for protease production, 12.5% were positive for HCN productions, 9.5 % were positive for cellulase production, 4% were positive for amylase production, 3.5% were positive for chitinase production. For abiotic stress tolerance test, all of the isolates were grown well at 20oc and 30oc and neutral pH, 27% isolates were grown well at 4oc, 25.5% grew at 40oc, 25.5% were grown well on pH-9 and pH-11, 23.5% were tolerated pH-5, 3.5% grew at 50oc and 60oc, 13.5% were grown well on 5% NaCl (w/v), 3.5% were grown well on 10 and 15% NaCl (w/v), which indicated these isolates can survive in some extreme conditions. Totally 15 bacterial species having PGP traits, biocontrol properties, and abiotic stress tolerance ability were identified using the Biolog bacterial identification system. Among these, the majority of the identified PGPR have utilized carbohydrate, carboxylic acid, and amino acid, which are the main components of plant root exudates. The above results indicated that thus PGPR can be used as biofertilizers as well as biocontrol agents to replace agrochemicals to improve crop productivity. Hence, these species can be further formulated and used for greenhouse and field applications.

Stable reference genes are indispensable for ensuring the fidelity of determined gene expression levels. However, the expression levels of reference genes are unable to remain constant under all possible experimental conditions. Therefore, the stability determination of reference genes is necessary in an experimental system set. In the preset study, the stability of nine cucumber candidate reference genes (CsACT, CsUBQ, CsEF1α, CsCYP, CsαTU, CsCACS, CsTIP41, CsYSL8 and CsHEL) subjected to stresses from Phytophthora melonis(P. melonis) were determined using four different analysis methods, including Delta Ct, BestKeeper, NormFinder and GeNorm. The study results revealed that CsUBQ and CsCYP were the most stable genes suitable as internal control in cucumber plants under attack by P. melonis condition.

Although laurel wilt disease was first reported in the United States in 2002 from redbay trees (Persea borbonia) around Savannah, Georgia it has rapidly spread throughout the southeastern coastal plain including Georgia and Florida. In the current study, transects were used to assess the spread and impact of the disease on two native bay trees redbay (P. borbonia) and swampbay (P. palustris) from north Florida in a semi-naturalized ecological preserve. Although tree size and mortality rates have been reported previously, this study provides the first size-based static life tables for both species. While a significantly higher percent (76%) of swampbay trees exhibited signs of laurel wilt disease compared to redbay trees (62%); redbay had more of its canopy damaged by the disease (41% vs. 32% for redbay vs. swampbay respectively); this resulted in a significantly smaller stem diameter for P. borbonia compared to swampbay, both species are experiencing significant declines due to the disease. Both species exhibited a Type III survivorship curve in which the vast majority of individuals were in the smallest size class (average stem diameter was only 2.5 and 3.6 cm for redbay and swampbay respectively). Although traditionally, population age (or size) structure that is heavily biased toward younger or smaller size classes suggests that the population is likely to expand in the future, for these bay trees high mortality rate due to beetle/fungal infestation of larger size classes is responsible for this trend; the smallest size classes are largely free from beetle infestation and laurel wilt disease because the stem diameter is likely insufficient to support beetle development. Results from this study suggest that swampbay is also highly susceptible to laurel wilt disease and its populations are likely to exhibit a similar (albeit slower) decline in Florida’s wetland and mesic ecosystems.

The circadian clock is an endogenous molecular oscillator with a period of about 24 hours, which regulates the physiology and developmental processes of almost all higher plants. Pseudo-response regulators (PRRs) are an important part of the central clock oscillator, together with other clock genes, constituting interlinked transcriptional feedback loops, which partly influence plant growth and development. In this study, a circadian clock-related gene MsPRR7 was cloned from Medicago sativa (alfalfa) by homologous cloning. The full length MsPRR7 gene was 2648 bp in length, with an open reading frame of 2385 bp encoding a protein of 795amino acids. Phylogenetic analysis showed that the MsPRR7 was closely related to PRR7 from the PRR family of Arabidopsis thaliana. Subcellular localization analysis found that MsPRR7 was located in the nucleus. Quantitative reverse-transcription polymerase chain reactions (qRT-PCR) demonstrated that expression of MsPRR7 gene transcripts in leaves was affected by circadian rhythms, and that its expression level increased with an extension of illumination time, reaching a peak around 8–10 hours. These results will provide the experimental basis for further study of the regulation of PRR family genes in alfalfa.

Pumpkins (Cucurbita pepo) are grown all around the world for a variety of reasons ranging from agricultural purposes to commercial and ornamental sales. The pathogens causing the rot of pumpkin in the world include fungi, bacteria, and viruses. The study was aim to identify fungal pathogens of pumpkin rot during storage, as well as control measures of the diseases using wood ash, mango leaf and rice chaff. Three hundred and sixty-six (366) fruits of pumpkins were studied in Pela, Gaya and Kulinyi districts of Hong Local Government Area of Adamawa State. The diseased samples (fruits) were randomly purchased. Of all the districts visited, Kulinyi has the highest percentage of disease samples (43.82%) while the least is Gaya district with 21.35%. Potato Dextrose Agar (PDA) was used for the isolation of pathogens and these gave Fusarium solani, Aspergillus niger, Aspergillus flavus, and Phytophthora capsici. All the fungal isolates exhibited different degree of pathogenic effect on the pumpkin fruits. The pathogens are susceptible to treatment both In-vitro and In-vivo control trials with wood ash and mango leaf at p ≤ 0.05. Inhibition improved with increased in concentration of the wood ash and mango leaf. Rice chaff treatment equally proved worthwhile with significant inhibition compared to the control at p ≤ 0.05.

The calyces of Hibiscus sabdariffa have been used by many communities as herbal tea. Their anthocyanin contents have been reported as the key component in anti-obesity studies. This present work reported results of anthocyanin content of calyces in two varieties of H. sabdariffa collected from Sabak Bernam, Selangor, Malaysia. The samples have been authenticated in the Herbarium, Institute of Bioscience, University Putra Malaysia prior to the study. The samples were processed and the ground dry raw material and its aqueous extract were analyzed using Fourier Transform Infrared (FTIR) and Two-Dimensional Infrared (2DIR). The short hybrid calyces (FT11-15A) raw material spectrum showed more than 80% similarity with long wild variety calyces (FT11-15B) when using “Compare” in analysis. The differences of both samples were obviously shown in their aqueous extract spectra. The peak at 1672 cm-1 and 841 cm-1 showed that tri-substituted double bond in FT11-15B aqueous extract was not present in FT11-15A aqueous extract spectra, whereby a double peak was assigned at 1221 cm-1 referred to anti symmetry stretching of aromatic and vinyl =C-O-C- with other =C-O- and 1192 cm-1 is assigned In-plane δ C-H in FT11-15A aqueous extract. The peak at 1071 cm-1 assigned as bonding C-H in plane bending of phenyl of both samples was the only peak comparable with standard delphinidin and cyanidin which are used for qualification and quantification of sample content. Aqueous extract spectra of both samples showed higher number of peaks detected compared with raw material spectra, which was attributed to the higher solubility of anthocyanins in water. The 2DIR correlation spectroscopy is advantageous in enhancing the qualitative analysis of herbal products. The anthocyanin content in both varieties of H. sabdariffa in descending amount is delphinidin-3-O-sambubioside (DS), cyanidin-3-O-sambubioside (CS), delphenidin-3-O-glucoside (DG) and lastly cyanidin-3-O-glucoside (CG). FT11-15A has more content of DS and DG of raw material and CG of water extract plus TFA than FT11-15B, whereby, FT11-15B has more content of CS, CG of raw material and DS, DG, CS of water extract plus TFA than FT11-15A.

Standardized method of seed treatment is of prime importance in the production of groundnut. The study was to carry out control trial using bark extract (aqueous and ethanol) and oil (seed) of mahogany (Khaya senegalensis) on seven (7) isolated fungi from two groundnut varieties (peruvian and valencia). The result shows that both mahogany bark and seed extracts are capable of inhibiting mycelial growth of all the isolates. There was no significant variation between the aqueous and ethanol bark extracts in-vitro, however the in-vivo test shows a significant difference between the aqueous and the ethanol bark extract in which the ethanol extract reduced growth of the pathogens more than the aqueous. For all the pathogens except Rhizopus stolonifer there was no growth between 50% to 100% concentration of the Khaya senegalensis oil in-vitro, however in-vivo control at 50% produced scanty to moderate growth for all the pathogens except Rhizopus stolonifer on peruvian, while there was full coverage on the seeds of valencia variety with Aspergillus niger and Rhizopus stolonifer having total coverage though Pseudaiiescheria boydii and Cylindrocarpon lichenicola were effectively inhibited and showed no growth at the 50% and 100%. Further research to focus on the quantifying the chemical constituents and formulation are suggested.

Cowpea plants naturally infected with cowpea mosaic comovirus (CPMV) showed different mosaic, mottle, dwarfing, and vain clearing symptoms. Diseased plants were ollected from certain locations of Alexandria and El-Beheira governorates during the growing seasons from 2011 to 2012. CPMV was detected in infected sap at 8 to 24 days after inoculation by DBIA, indirect ELISA and tissue blot immunoassay (TBIA). Chlorotic local lesions were observed on Chenopodium amaranticolor in infectivity test. By using indirect ELISA and DBIA, CPMV were detected in infected plant sap of serial dilutions up to 1: 400. The incidence of CPMV in 21 day old cowpea seedlings grown from infected seeds was determined by ELISA and positive detection of virus antigen reached 65%. Nitrocellulose membrane and canson paper could be used as solid carriers in TBIA and DBIA for detection of CPMV in infected plant tissues. Results revealed that both faces of nitrocellulose membrane and canson paper could be used as solid carriers in TBIA for detection of CPMV in infected plant tissues. According to reverse transcription polymerase chain reaction (RT-PCR) assay of CPMV infected plant; the amplified product was approximately 800bp of partial coat protein gene. The nucleotide sequences accession number were LN606585 and LN606586. The phylogenetic tree was generated using sequences of CPMV isolates with the other CPMV records from GenBank.

Goosegrass (Eleusine indica L. Gaertn.) is a troublesome weed in turfgrass systems throughout the world. The development of herbicide resistant ecotypes has occurred to multiple modes of action. Goosegrass is a prolific seed producer (~50,000 per plant), fast growing and diverse weed. Such growing attributes make it essential to have a better understanding of the genetic diversity of various ecotypes. The objectives of this study were to determine if morphologically distinct goosegrass ecotypes collected in Florida were phenotypically distinct and genetically different. Phenotypically, the goosegrass ecotypes can be classified as follows; dwarf, intermediate 1 (int_I), intermediate 2 (int_II) and wild. The dwarf had the least seedheads followed by the wild ecotype; 5 and 17 respectively, while int_I and int_II had highest number of seedheads; 22 and 34 respectively. The dwarf ecotype had lowest height of 6 cm and the wild ecotype had highest height of 36 cm. Dwarf and int_II ecotypes had shortest internode length of 0.2 cm and 1 cm, respectively, while the wild ecotype had longest internode length of 7 cm. The dwarf ecotype had lowest number of racemes per plant of 1, while the wild ecotype had highest number of racemes per plant of 7. Total biomass was lowest for the dwarf and int_II ecotype; 0.7 g and 1.5 g, respectively, and total biomass was highest for the wild ecotype at 5 g. Gene sequencing of two rice (Oryza) gene sequences (accession AP014964 (gene A) and AP014965 (gene B)) and subsequent phylogenetic analysis suggest the ecotypes are genetically different. Three single nucleotide polymorphisms (SNP) of interest were discovered indicating allelic differences between ecotypes.

The use of enzyme linked immunosorbent assay (ELISA) for the detection of plant viruses is well documented. It proved to be a very valuable detection tools for the plant viruses. The efficiency of the ELISA technique was for practical purpose independent of the ratio of antibodies to antigen. This avoids the necessity of making specific enzyme conjugates for each antigen to be tested and eliminates the extreme specificity, thus allowing for quantitative evaluation of strain relationships. The advantages of indirect ELISA are sample. It needs only to be macerated and added to the plate. The crude antiserum could be used, although it should be cross absorbed before to prevent spurious host reaction. Single commercially available second antibody conjugate is utilized, thus eliminating the problems of preparing and storing many different conjugated antisera. Blotting technique has become widely used for specific identification of nucleic acid and proteins. This dot assay was modified to detect protein by spotting the antigen on a nitrocellulose membrane and incubating the membrane in test antibody followed by incubation in peroxidase-conjugated second antibody to the first antibody, and by development in 4-chloro-1-naphthol. The above procedure termed dot blot immunobinding assay (DBIA). The technique of tissue blotting on nitrocellulose membrane was described for detection of plant viruses in infected plants. Tissue blots were made by pressing with a firm and gentile force, the freshly cut tissue surface on nitrocellulose membranes. The possibility of using both sides of the nitrocellulose membrane (NCM) by tissue blot immuno assay (TBIA) for the detection plant viruses. In an effort to reduce the cost of virus assays, different types of regular paper were evaluated as possible replacements for the commonly used nitrocellulose membrane (NCM) as the solid phase in the tissue-blot immunoassay (TBIA) were used. Comparisons between different serological methods were demonstrated by many investigators Dot immunobinding was eight times more sensitive for detection of PVX and four times more sensitive for detection of PVS and PVY than DAS-ELISA.

Physicochemical and pharmacological studies indicated that Filicium decipiens seeds contained various specialized metabolites, including saponins. The aim of this work is to reveal the nephrotoxicity of FDS, a saponin isolated from Filicium decipiens seeds on male Wistar rats histopathological and biochemical parameters. Rats were submitted to oral ingestion of FDS (6.0 mg/kg) and crude extract (120.0 mg/kg) and were observed high levels of urea and creatinine in blood analyses of all animals followed by an acute renal failure by glomerular retraction. In the present study, FDS and crude extract when administered in Wistar rats induced an increase of serum levels of Urea and Creatinine, biochemical markers of kidney function. Table 1 shows Urea concentration at Test group with FDS (54.3 ± 1.80 mg/ml) and Test group with crude extract (49.7 ± 2.00 mg/ml), were 47% and 34.7% higher, respectively, when compared to control group (36.9 ± 2.00 mg/ml), and Creatinine at the test group with FDS (2.1 ± 0.03 mg/ml) and test group with crude extract (1.6 ± 0.09 mg/ml) presented a value 3.5 and 2.8 times higher, respectively, than control (0.6 ± 0.08 mg/ml). Based on these results, our data demonstrate a significant effect in renal function of rats treated with F. decipiens saponin.