with

Agrobacterium rhizogenes ATCC 15834 wild type strain was transformed with the binary vector pBI121 using the heat shock method. The transformed Agrobacterium was then tested for virulence through tobacco leaf explant transformation. Compared to the non-transformed Agrobacterium, the transformed Agrobacterium showed reduced virulence, producing significantly lower number of hairy roots in tobacco leaf explants. Although the transformed Agrobacterium showed reduced virulence, it was able to transfer the T-DNA of the binary vector into the plant genome, resulting in stable GUS expression in the generated hairy roots. This indicated that in addition to the transfer DNA (T-DNA) from its root inducing (Ri) plasmid, the transformed Agrobacterium is also capable of transferring the binary vector T-DNA and allowing the integration of a foreign gene. Results also showed that hairy root generation efficiency of the transformed Agrobacterium varied with the concentration of the selection agent (kanamycin). Hairy root generation efficiency (hairy roots·explant-1) progressively increased with decreasing concentrations of kanamycin; and the efficiency was highest in the absence of kanamycin. Generated hairy roots showed very strong to tiny GUS expression even those that grew under the highest concentration of the kanamycin (50 mg·L-1). This indicated that co-transformation and efficient transgene expression does not always occur.

Vochysia haenkeana extract (Vh-E) was assessed against the neuromuscular blockade induced by Bothrops jararaca venom on chick biventer cervicis (BC) preparation. Pre- and post-venom incubation treatments (Pre-vit and Post-vit) were analysed here. Contractures ACh (110 µM) and KCl (20 mM) were evoked before and after addition of venom without stimulation. Vh-E (600 µg/mL) under Pre-vit was more efficient to neutralize the neuromuscular blockade by venom (40 µg/mL) [72.5±4.6% (venom) vs. 45.2±14% (Vh-E) of blockade, p<0.05, n=4]. Vh-E (600 µg/mL) did not cause significant changes under Post-vit [72.5±4.6% (venom) vs. 63.4±8.2% (Vh-E) of blockade, n=4]. The Pre-vit inhibited the blockade of the contracture to ACh (106±17% of response; n=4) while the Post-vit was able to attenuate the effect of the venom on this contracture (55±5% of response; n=4); related to those contractures to KCl both of treatments with Vh-E attenuated the blocker effect of the venom (62.5±7.7% and 55±5% of response for Pre-vit and Post-vit, respectively; n=4). In conclusion, Vh-E neutralizes partially the neuromuscular blockade in Pre-vit, an effect that can be related to preserved function of “extrinsic” post-synaptic receptors, by measured contractures in response to ACh. The myotoxicity of the venom was significantly reduced by Vh-E in both, Pre-vit and Post-vit, by measured contractures in response to KCl.

Hepatitis B virus (HBV) is one of the world’s major infectious diseases with 350 million people who are chronic carriers of HBV [1]. Significant minorities go on to develop liver cirrhosis or hepatocellular carcinoma and over 1 million die annually from HBV-diseased liver. Janahi E. at faculty of science, Bahrain University, Bahrain has submitted the following information [2], on HBV-genome organization as part of his Ph.D. degree (2007) in Imperial College, England. HBV genomic organization has 4 Open Reading Frames (ORFs) i.e. Pre-S/S Gene, Pre-C/C ORF, P ORF and X ORF. Regulatory Elements has 4 promoters (pre S2, pre S1, C promoters and X promoters), Pregenomic RNA, Enhancers (Enh 1 and Enh 2) where they are involved in cccDNA formation, Glococorticoid-Responsive Element which is located in X ORF and P ORF overlapping, Polyadenylation Signal (Direct Repeat 1 (DR1) and Direct Repeat 2 (DR2)), Epsilon-Stem Loop and Post-Transcriptional Regulatory Element. HBV genotype D is prevalent in our Middle East area. The HBV genome is a partially relaxed-circular dsDNA molecule consisting of a full length strand (minus strand) with a single unique nick and a complementary (positive strand) of variable length. HBV is considered as a para-retrovirus because its replication involves the reverse transcription of an intermediate-RNA function, of pre-genomic RNA (pgRNA). Replication of HBV genome starts with the encapsidation of the pgRNA and encodes HBV polymerase into an immature nucleocapsid formed by the viral core antigen.

Capsicum spp. is one of the most important economical horticulture crops due to its high consumption either by fresh vegetable or dried spice. Molecular genetic markers offer a number of applications in the genetic improvement of crop plants, which plays an important role in the areas of plant classification and breeding programs. The polygenetic characters of rare species, which are difficult to analyze by traditional methods can, be analyzed easily and classified by using molecular markers. In our study, genetic relationships of twenty-two paprika species were examined to estimate their genetic variations/similarities and to detect the polymorphism present within and among the paprika species by using RAPD-PCR markers. The results revealed that the maximum similarities among the 16 ICBD lines were 100%. The ICBD 03 had 76% similarity compared with other ICBD lines. The CC01 had comparatively low similarity with ICBD forms (30%), followed by EC01 (28%), EC02 (33%), CC02 (35%), and Kt.Pl-19 (60%). The similarity between EC01 and EC02 were 54%. Kt.Pl-19 showed different similarities compared to CC01 (41%), CC02, EC01 (38%), EC02 (29%) and ICBD 03 (40%). The different combinations were tried to optimize the RAPD-PCR profile, which helped to assessing the polymorphism/similarities within and among the Paprika germoplasms were studied.

Natural dyes have become a viable alternative to expensive and rare organic sensitizers because of their low cost, easy attainability, abundance of supply of raw materials and environmental friendliness. Chlorophyll, the most abundant pigment, can be extracted from plant leaves with simple and inexpensive methods, but it’s difficult to use as a Dye-Sensitized Solar Cells (DSSC) sensitizer due to the absence of OH and COOH groups. The opposite is true for xanthophylls, a particular class of carotenoids that contain free hydroxyl groups and thus may be considered as potential DSSC sensitizers. In this work we describe a new and inexpensive method of chlorophyll extraction from leaves based on the use of a basic solvent that provides the creation of COOH groups, allowing chlorophyll binding on the TiO2 layer. This modified chlorophyll dye showed a higher DSSC efficiency level (0.72%) compared to xanthophylls, which had lower efficiency.

Vegetable production by grafting is a technique which it has made possible to resume agricultural soils which previously could not be produced due to stress generated by various abiotic factors, like a lack of water, stress by high or low temperatures, and or heavy metal contamination, among them. It has been documented and defined a number of graftings which they are tolerant to different factors; however, when it comes to auscultating information related to understand the molecular responses and observe what are the biochemical changes and physiological responses of grafted plants, it is dispersed. The current paper attempts to provide basic information documented on technique, addressing the molecular, biochemical and physiological responses, and thus get a clear perspective on the use of grafts, making this practice be used with most frequently by all its advantages.

Psidium guajava (guava) is an important crop and economic resource in many tropical countries and Brazil stands out as one of its major commercial producers. The guava crop has been severely attacked by the plant parasitic nematode Meloidogyne enterolobii which has caused drastic reduction of productivity and, in some cases, even total loss of cultivated areas. The use of resistant rootstocks that are tolerant to these soil pathogens can be a low-cost solution to this pathogen, which has established itself as one of the major constraints for the cultivation of guava in Northeast Brazil. The objective of this work was to evaluate the reaction of P. guajava and P. guineense to M. enterolobii infection under greenhouse conditions, with an infection period of 60 days. 12 P. guineense, and 4 P. guajava were evaluated. The host response to the nematode infection was evaluated according to the following parameters: gill index (GI), reproductive factor (RF), and reduction of reproductive factor (RRF). Considering the RRF criteria, the P. guineense genotypes, AR1, AR3, AR4 and AR10 were considered resistant. The RF index was considered inadequate to evaluate resistance in the 60 days period, due the slow development of the parasite in P.guineense.

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.

The armored scale Aulacaspis alisiana, is a serious invasive pest of the Japanese silver tree, Neolitsea sericea, causing serious damage to the tree in Japan. However there are currently no control approaches available for it, complicated by shortage of information on the pest. We studied life history strategies of A. alisiana on N. sericea in Fukuoka Prefecture with a view to providing a basis for formulating sustainable control based on an understanding of the behavior of the pest and potential role of its natural enemies. We established that A. alisiana had three overlapped generations in Fukuoka, with generation times ranging between 65 and 71 days. The adults were relatively fecund, with each female producing between 60 and 67 eggs, with high hatchability, >78%. The pest settled on the lower side of leaves, and although it generally preferred younger leaves, it did not attack newly emerged leaves. Natural enemy groups comprising ants, spiders and beetles (coccinelids) played an important role in regulation of the pest population, with natural mortality of about 30%. They could thus form a critical component of an integrated management approach for the pest in Fukuoka.

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.

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.

Peaches, Prunus persica were planted as grafted saplings in an avocado orchard previously infested with Armillaria mellea (Vahl) P.Kumm. Trees were planted in large or small holes with or without fresh yardwaste chips added as an amendment and with or without a Trichoderma biocontrol product sprayed into the hole. Trees were monitored for six years -- growth and mortality was tabulated. Six years later 40% of the trees had died from the disease. Trees planted in a large hole were more likely to survive than in a smaller hole (P=0.07) and trees in large holes with fresh organic matter added were the most likely to survive (P=0.04). Trichoderma sprays in the planting hole did not increase survival rates. While growth was initially retarded by adding fresh yardwaste to the hole, in later years none of the treatments affected growth rates.

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.