Constructional and Functional Evaluation of Two New Plant Expression Vectors—pBI121gus-6 and pBI1215+1

DOI: 10.7904/2068-4738-IX(17)-60

Reza MOHAMMADHASSAN1, Kasra ESFAHANI2*, Bahareh KASHEFI3

 1College of Agriculture, Damghan Branch, Islamic Azad University, Damghan, IRAN

2National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, IRAN

3College of Agriculture, Damghan Branch, Islamic Azad University, Damghan, IRAN

Corresponding Author: kasra13@gmail.com, Phone: +982144787451

Abstract. Binary vectors are widely used in Agrobacterium gene transfer in plants. Although new plant expression vectors have been designed, pBI121–based vectors are still more common. The availability of fewer numbers of restriction enzyme sites is the most important drawback in pBI121. In this article, two new vectors–based on pB121, pBI1215+1, and pBI121GUS–6–were introduced. The construction of new vectors was confirmed by PCR and digestion pattern analysis. Furthermore, the reporter gene (gus) was cloned in these vectors. The T–DNA transformation ability of the new vectors and pBI121 (as the control sample) to tobacco via Agrobacterium tumefaciens strain LBA4404 was evaluated. Transgenic plants were regenerated with BAP and NAA in a selective medium. Thereafter, plant DNA was extracted and successful gene transformation was confirmed by PCR analysis. The GUS assay also confirmed gene expression in transgenic leaves. The results indicate that the vectors are totally efficient in plant transformation.

Keyword: Agrobacterium–mediated transformation, binary vector, Plant expression vector, pBI121, pBI1215+1, pBI121GUS–6

Toxicity and repellency of three Algerian medicinal plants against pests of stored product: Ryzopertha dominica (Fabricius) (Coleoptera: Bostrichidae)

DOI: 10.7904/2068-4738-IX(17)-50

Kada RIGHI1*, Fatiha ASSIA RIGHI1, Ahlem BOUBKEUR1, Karima BOUNGAB1, Abdelkader Elouissi 1, Ali Cherif DJENDARA2

 1Laboratory of Research on Biologic systems and Geomantic. Department of Agronomy Sciences. Faculty of natural Science. University Stambouli Mustapha Mascara. ALGERIA.

2Laboratory of Physical Chemistry of macromolecules and Biological interfaces. University Stambouli Mustapha Mascara. ALGERIA.

Corresponding author righika@yahoo.fr. Tel. +213 772563022

Abstract. In order to find alternatives to chemical synthetic insecticides in the fight against pest of stored products, we evaluate the insecticidal and repellent activities of three essential oils by contact on the adults of R. dominica. Oils were extracted by hydrodistillation technique for 4 h using a Clevenger from the indigenous medicinal plants then the identification of the constituents was carried out on the basis of gas chromatography coupled with the GCMS spectrometry type Shimadzu TQ8030 coupled with a spectrometer. The different plants are S. molle, M. rotundifolia and S. calamintha. Our results of the contact–toxicity test of oils on substrate showed that the essential oil of M. rotundifolia has a highly significant insecticidal effect with a 100 % mortality at a concentration of 3 μL from the first h of treatment, Similarly the essential oil of S. calamintha has a highly significant biocidal effect with a 100 % mortality at a concentration of 5 μL even the first h of treatment also, the essential oil of S. molle that has an insecticidal effect with a mortality of 100 % at a concentration of 25 μL after 96 h of the treatment. The results of the study of the repulsive effect of the oils tested against the insect showed that the three essential oils possess a remarkable repelling effect just after 2 h of exposure with a PR (The percentage of repulsion) of 50 % for S. molle, 100 % for S. calamintha and M. rotundifolia. The toxicity test of dry powders against the adult beetle showed that the powders exerted their biocidal effect (1.2 g) with a mortality rate of 96, 90 and 75 % respectively for S. calamintha, M. rotundifolia and S. molle after 96 h of treatment. Biotechnological control through the use of essential oils extracted from these plants is tested as a bio insecticide has proved very effective in the protection of cereals against such insects that degrade quality and reduce yield and in addition threaten health of the consumer.

Keyword: Schinus molle, Mentha rotundifolia, Satureja calamintha, Ryzopertha dominica, cereals, toxicity.

 

Possibility for weed control by using of an organic product with herbicidal effect

DOI: 10.7904/2068-4738-IX(17)-40

Natalia Georgieva1*, Ivelina Nikolova1, Yordanka NAYDENOVA2

 1Department of Technology and ecology of forage crops, Institute of Forage Crops, Pleven, BULGARIA

2Department of biochemistry, Institute of Forage Crops, Pleven, BULGARIA

Corresponding author: e–mail: imnatalia@abv.bg

Abstract. The possibility for weed control in noncropped areas (stubbles) by using an organic product with herbicidal effect (Segador), alone and combined with low dose of synthetic herbicide (Roundup, 360 g/L glyphosate), was investigated. The individual application of Segador at a dose of 18 l/ha suppressed completely the annual grassy weeds and also showed a very good effect against broadleaf weeds and perennial grassy weeds (85–95 % efficacy), therefore could be recommended for organic farming. The combined application of Segador with Roundup at different doses significantly increased the efficacy of the herbicidal mixture. Its application depended on the predominant weed species in the cultivated area. The combination of Segador and Roundup, each one at a dose of 6 l/ha, provided good control of annual weeds and Sorghum halepense, but a certain resistance showed some perennial broadleaf species (Convolvulus arvensis, Rumex crispus and Cynodon dactylon) as the efficacy was in the limits 75–97 %. Increasing the dose of Segador up to 12 L/ha in combination with Roundup 6 L/ha suppressed Rumex crispus and Cynodon dactylon and increased the efficiency in Convolvulus arvensis to 95 %. Full control of annual and perennial weeds, provided the organic product Segador 18 L/ha + herbicide Roundup 6 L/ha, as the effect of the mixture was equalized with that of Roundup at a dose of 12 L/ha. The low dose of application of Roundup in combination with different doses of Segador minimized the use of synthetic herbicide and is recommended for conventional production.

Keyword: bioherbicide, weed control, efficacy, noncropped areas.

 

Characterization of the quality of the steamed yoghurts enriched by dates flesh and date powder variety H’loua

DOI: 10.7904/2068-4738-IX(17)-31

Ahmed HARIRI1*, Nawel OUIS2, Djilali BOUHADI1, Zouaoui BENATOUCHE1

 1Bioconversion Laboratory, Microbiology Engineering and Health Saftey, University Mustapha Stambouli of Mascara (UN 2901), BP. 763, Sidi Said, 29000, Mascara, ALGERIA.

2Laboratory pf Physical Chemistry of Macromolecules and Biological Interfaces, University Mustapha Stambouli of Mascara (UN 2901), BP. 763, Sidi Said, 29000, Mascara, ALGERIA.

Corresponding author*: haririahmed@yahoo.fr. Tel. 00213661721190

Abstract. The purpose of the present work was to investigate the effect of including date flesh and date powder in the steamed yoghurt processing on some quality during four weeks of storage. In order to obtain the powders, date variety of H’loua was washed, cut into small particles, and then dried at 90 °C during 2 h. Seven steamed yoghurts were manufactured: control prepared by 80 g/L sucrose, yoghurts manufactured by replacement of some quantity of sucrose by 10, 20 and 25 g/L of the date powder and yoghurts prepared by replacement of some quantity of sucrose by 60, 70 and equal quantity 80 g/L of date flesh. All samples were analyzed for some physicochemical, biochemical, microbiological and sensory characteristics at 1, 7, 15, 21 and 29 days of storage. The total or partial replacement of the sucrose by date flesh and date powder improves the nutritional quality of the yoghurts such as proteins, fat, ash, and dry mater, but the level of the total sugars was decreased. Results of microbiological analyze showed complete absence of the total and fecal coliforms, Staphylococcus aureus, faecal streptococci and Salmonella in all yoghurts prepared and throughout the storage period. Replacement of the sucrose by all types of date improves the sensory characteristics of the yoghurts included taste and texture.

Keyword: Date flesh, date powder, H’loua, Quality, Sucrose, Yoghurt.

Antixenosis Resistance in Sugar Beet Varieties to Long Snout Weevil Lixus incanescens Boh. (Col.: Curculionidae)

DOI: 10.7904/2068-4738-IX(17)-24

Faramarz HARIRI MOGHADAM1*, Jafar KHALGHANI1, Saeid MOHARRAMIPOUR2, Babak GHARALI3, Mehrzad MOSTASHARI MOHASSES3 

1Department of Entomology, Science and Research Branch, Faculty of Agriculture, Islamic Azad University, Tehran, IRAN

2Department of Entomology, Faculty of Agriculture, Tarbiat Modares University, Tehran, IRAN

3Agriculture and Natural Resources Research and Education Center of Qazvin Province, Qazvin, IRAN

*Corresponding author: Fhmoghadam@gmail.com, Tel: +989127852004

Abstract. In plants, antixenosis resistance could reduce injury and yield loss; they can produce selection pressures on insect herbivores that lead to pest resistance. Traditionally, sugar beet long snout weevil, Lixus incanescens Boh. (Col.: Curculionidae), is one of the serious pests causing sugar beet losses. In the present study, antixenosis resistance was evaluated in 11 varieties of sugar beet including Pars, Torbat, Ekbatan, SBSI019, Rizofort, Puma, Dorothea, Brigita, Pecora, Nagano and Jolgehin a field trial from 2013 and 2014 years in Qazvin province, Iran. A randomized complete block design with their replications was used. The results showed that in both years, in the first 24 h experiment after release the lowest and highest densities of pest (No. adult/plant) were seen on Pars (2.99) and Dorothea (2.83) varieties, respectively, as opposed the lowest antixenosis resistance were found on Brigita (8.66) and Nagano (8.83) cultivar, whereas on other cultivars showed intermediate values. In Pars, Brigita and Nagano varieties the number of adult weevils (adult/plant) increased rapidly during the first 24 h, but after which there was a decline to the minimum on about the fifth day. The results of data analysis to investigate the mechanism of free choice antixenosis in the years 2013 and 2014 showed that there are no significant differences between experimental blocks and among cultivars tested. As a result, our findings showed that above mention variety can be used in integrated management plans of sugar beet long snout weevil, and with regionally adapted varieties and excellent seed quality we strive for excellent varietal performance in all growing regions of the Iran.

Keyword: Antixenosis, Lixus incanescens, Resistance, Sugar beet, Weevil.

Antioxidant and antibacterial activities of the essential oils of Ceratonia siliqua

DOI: 10.7904/2068-4738-IX(17)-13

Nawel OUIS1*, Ahmed HARIRI2

 1Laboratory of Physical Chemistry of Macromolecules and Biological Interfaces, Faculty of Science the Nature and Life, University Mustapha Stambouli of Mascara (UN 2901), BP. 763, Sidi Said, 29000, Mascara, ALGERIA

2Bioconversion Laboratory, Microbiology Engineering and Health Safety, Faculty of Science the Nature and Life, University Mustapha Stambouli of Mascara (UN 2901), BP. 763, Sidi Said, 29000, Mascara, ALGERIA

*Corresponding author: nawel_chim@yahoo.fr

Abstract. The present study investigates the antioxidant and antibacterial activities of the essential oils obtained by hydrodistillation of the pulp and the seeds of Ceratonia siliqua. Dried pods (pulp or seeds) were subject to the hydrodistillation for 3 h using a Clevenger–type apparatus. The two essential oils obtained were characterized by its physicochemical indications. Theses oils were used for the determination of the antioxidant activity by free radical scavenging activity, reducing power and liver lipid peroxidation methods and for the evaluation of the antibacterial properties against referenced tested bacteria. The oil of the pulp presents a high scavenging power of free DPPH radical 89.0 ± 1.2 % at concentration 1000 µg/mL and IC50 7.8 µg/mL compared to the seeds 79.0 ± 1.6 %, IC50 31.25 µg/mL. A significant activity for reducing iron and protection against lipid peroxidation induced by Fe+2 were obtained by the oil of the pulp. The essential oils of carob showed a good antibacterial activity against tested referenced pathogenic bacteria Staphylococcus aureus ATCC 25923, Pseudomonas aerugenosa ATCC 25853 and Escerichia coli ATCC 25922.

Keyword: Antibacterial, Antioxidant, Carob, Essential oils, Pulp, Seeds.

Investigation of the induced antibiosis resistance by zinc element in different cultivars of sugar beet to long snout weevil, Lixus incanescens (Col: Curculionidae)

DOI: 10.7904/2068-4738-IX(17)-5

Faramarz HARIRI MOGHADAM1*, Jafar KHALGHANI1, Saeid MOHARRAMIPOUR2, Babak GHARALI3 and Mehrzad MOSTASHARI MOHASSES3

 1Department of Entomology, Science and Research Branch, Faculty of Agriculture, Islamic Azad University, Tehran, IRAN

2Department of Entomology, Faculty of Agriculture, Tarbiat Modares University, Tehran, IRAN

3Agriculture and Natural Resources Research and Education Center of Qazvin Province, Qazvin, IRAN

*Corresponding author: Fhmoghadam@gmail.com, Tel: +989127852004

Abstract. Sugar beet weevil, Lixus incanescens Boh., is one of most dangerous pests in sugar beet. Host plant resistance is known as an efficient strategy for pest management. Thus, this study was conducted to investigate the antibiosis resistance induced by zinc element in different cultivars of sugar beet to long snout weevil, L. incanescens. Seeds of eleven commercial monogerm cultivars of sugar beet including Pars, Torbat, Ekbatan, SBSI019, Rizofort, Puma, Dorothea, Brigita, Pecora, Nagano and Jolgeh, were studied. Field was divided to two parts; one part was treated with zinc fertilizer and other part did not treat with fertilizer. Antibiosis parameters were tested. Results clearly did not show that cultivars have antibiosis resistance under non–using fertilizer during 2013 and 2014 years. Rizofort cultivar showed lower susceptibility compared with other cultivars when using zinc fertilizer, because of lower adult weight, female longevity and period length of immature stage during the both years. Thus, Rizofort cultivar was relatively resistant to L. incanescens when using zinc fertilizer during the both years, which can be useful in the development of IPM programs for this insect in sugar beet fields

Keyword: Lixus incanescens, antibiosis, sugar beet, zinc fertilizer.

Study of genetic diversity in local rose varieties (Rosa spp.) using molecular markers

DOI: 10.7904/2068–4738–VIII(16)–148

Abbas SAIDI1*, Yazdan EGHBALNEGAD1 and Zahra HAJIBARAT1

1Department of Plant Sciences and Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, G.C, Tehran, IRAN

Corresponding author: abbas.saidi@gmail.com, Phone: +982129901964

Abstract. This study was undertaken to evaluate genetic diversity in a germplasm consisting of rose varieties. Genetic distances were estimated using three different molecular marker techniques including: start codon targeted (SCoT), conserved DNA–Derived Polymorphism (CDDP) and directly amplified minisatellite DNA (DAMD). According to the results, the average polymorphism information content was 0.37, 0.36, and 0.36 for SCoT, CDDP and DAMD markers, respectively indicating that the studied marker types were equal in terms of assessing diversity. Cluster analysis using SCoT and CDDP divided the varieties to four distinct clusters whereas DAMD markers data, grouped the varieties into three clusters. There was a positive significant correlation (r=0.92, p<0.01) between similarity matrix obtained by SCoT and CDDP. Results suggested that the efficiency of SCOT, CDDP and DAMD markers had a relatively same efficiency in fingerprinting of varieties. This is the first time that the efficiency of the three molecular markers have been compared with each other in a set of rose samples. The results showed that the studied markers had an appropriate polymorphism and thus were suitable for the study of genetic diversity in rose.

Keyword: Fingerprinting, PIC, molecular marker, genetic distance, correlation.

Alkaline protease producing Bacillus isolation and identification from Iran

DOI: 10.7904/2068–4738–VIII(16)–140

Maryam ARAMESH, Hatef AJOUDANIFAR*

Department of Microbiology, Damghan Branch, Islamic Azad University, Damghan. IRAN

*Corresponding Author: Email:ajoudanifar@gmail.com, Tel: +989124023012

Abstract. Although several microorganisms are known to produce proteases, Bacillus strains are often preferred as major sources for commercial alkaline proteases due to their exceptional ability to secrete large amounts of highly active enzymes. The present study aimed at isolation and molecular identification of alkaline protease producing Bacillus spp. from soil samples of Damghan County in Iran. This research was conducted in the laboratory complexes in the school of pharmacy of Islamic Azad University Damghan Branch. Soil samples were collected from 3 different regions. Streaking plate was done and 9 colonies were selected on the basis of appearance. To identify isolates, biochemical tests were performed. 16SrRNA gene duplicated with PCR and sent to Sinagene Company for sequencing. All isolates were oxidase–negative and catalase–positive. The ONPG tests were negative. 16SrRNA analysis showed that the isolated bacteria belonged to genus Bacillus. The results confirmed that four bacterial isolates had alkaline protease production capacity. It appears that after complementary examinations, the isolated Bacillus strains can be used for production of alkaline protease.

Keyword: alkaline protease, Bacillus, enzyme, biochemical tests.

Effect of Intravenous Infusion of Hypertonic Glucose on the Treatment of Cows with Postpartum Anestrus

DOI: 10.7904/2068–4738–VIII(16)–133

Seyed Fayaz MIRASI OSKOEE1, Hossein HAMALI*2, Reza ASADPOUR2

1Graduated student, Faculty of Veterinary Medicine, The University of Tabriz, IRAN

2Department of Clinical sciences, Faculty of Veterinary Medicine, The University of Tabriz, IRAN

*Corresponding Author: E–mail: hhamali@yahoo.com; Phone: +98 914 114 5575

Abstract. The objective of this study was to evaluate the efficacy of intravenous infusion of hypertonic glucose for recycling of the cows with postpartum anestrus. One hundred true anestrus Holstein–Frisian cows with ≥ 60 days in milk (DIM) according to the ultrasound examinations were selected from a dairy herd located on the suburb of Tabriz. Anestrus cows randomly were allocated into two groups: A&B (n=50). In group A (Gluco–Select synch), cows received 500 mL hypertonic glucose for 3 consecutive days (–3 to 0) IV, 0.021 mg bucerelin acetate on the day 0, IM followed by 25 mg dinoprost tromethamine IM on the day 7. In group B (Select synch), the same protocol was carried out on the cows except that the glucose hypertonic solution replaced by normal saline as placebo. Thereafter all cows were detected for estrus signs for the next 5 days and inseminated based on visual signs. Twenty seven out of 50 cows (54 %) in group A and 8 out of 50 cows (16 %) in group B showed the estrus signs respectively. Conception rates were recorded as A=44.5 %, B=37.5 % for the first insemination and A=B=100 % for the second insemination of cows. The number of insemination per pregnancy was calculated as 1.55 and 1.62 for groups A and B respectively. In the case of estrus rate, the difference between two groups was statistically significant (P≤0/01), but in the cases of conception rate or the number of insemination per pregnancy rate the differences were not statistically significant.

Keyword: Anestrus, Cow, Glucose, Hypertonic, Postpartum.