PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY

Understanding nutrient content and biomass accumulation is important for fertilization schemes, harvesting management, and sustainable plantation management to meet demands on timber and fuel. In this study, biomass and nutrient accumulations were studied for Prunus arborea trees of 1- to 5-years old. Medium-sized trees were sampled to collect biomass and representative samples were collected for nutrient (N/nitrogen, P/phosphorous, and K/potassium) analysis. The results indicated that power function was best fitted for a relationship between age and dry biomass of individual trees. While the polynomial pattern was best fitted for the relationship between ages and ratios of below- to above-ground biomasses with the lowest ratio of 21% in trees of between 3- and 4-years old. Among the four analyzed organs, N, P and K contents were highest in leaves and lowest in the stem. While that in branches and roots were similar. In leaves, N and K contents were 0.4–1.48%, while P contents were 0.10–0.36%. In stems, N and K contents were 0.15–0.61%, while P contents were 0.01–0.21%. It is concluded that low P contents may indicate the P deficiency in the site, leading to fertilization requirements. While N and K application may not be necessary. Remaining leaves in the site at logging may partly compensate soil nutrients from high leaf biomass (up to 26% total biomass) and the highest nutrient contents (up to 1.48%).

Lipases are the class of enzymes that hydrolyze the triglyceride and convert them to glycerol and fatty acids. This study provided the bioinformatics information about the Rhizobium leguminosarum Lipases. Physio-chemical properties were presented in details for different strains of Rhizobium leguminosarum. The total numbers of different strains of Rhizobium leguminosarum lipase’s residues were ranged from 504 (A0A072C2T8) to 208 (A0A072C300). Molecular Weight (MW) of different lipases were from 53657.02 (A0A072C2T8) to 22571.93 (A0A072C300). Rhizobium leguminosarum lipases are hydrophilic, soluble and cytoplasmic proteins. Numbers of Cys residues in different proteins were 3 to 5 however none of them forms the disulphide bond in the structure of lipases. The Isoelectric Points (pI) of the lipases were from 9.39 to 5.18 which represented the basic to acidic properties. Extinction coefficient (EC) of proteins were from 11460 (A0A072BXE4) to 46410 (A0A072C545) M-1.cm-1. The homology modelling of all 8 lipases were presented based on Swiss-model server. The evaluations of models were performed with PROCHECK program. These structures will provide valuable information for functional analysis of microbial lipases and can help in design the platform for engineering this enzyme.

Bacillus thuringiensis (Bt) is a ubiquitous Gram-positive bacterium that can produce different insecticidal proteins during the sporulation phase growth. The objective of this study was to examine the expression, including the effects of induction temperature, time and IPTG concentrations as well as investigate insecticidal activity of Cry8Db protein against Lepidiota signata Fabricious. The results showed that the cry8Db gene was expressed in Rosetta–gamy Escherichia coli strain at optimal temperature 28°C, 100 μM IPTG and for 4 h induction. SDS-PAGE and Western blot were applied to confirm the normal expression and transcription of the cry8Db gene which produced the polypeptide with a molecular mass of 73 kDa. Three stages of Lepidiota signata Fabricius larvae were examined in the bioassay to investigate their survival after 15 days. The protein exhibited high toxicity against Lepidiota signata Fabriciusin the three different larvae stages at the lowest mean lethal concentration of LC₅₀ =183.7 ng/mL, 270.8 ng/mL and 345.5 ng/mL, respectively. This is the first report demonstrating Cry8Db protein against Lepidiota signata Fabricius larvae. The Cry8Db protein may become a potential environmentally friendly marker for the biological management of Lepidiota signata Fabricius.

A field study was conducted during rabi season of 2015-16 and 2016-17 to study the effect of micronutrients and growth regulators on the pigment (chlorophyll and carotenoids) content, nitrate reductase activity and relative leaf water content of chilli. The treatments of the study comprised of five levels each of micronutrients including control (no micronutrient), two levels of ZnSO₄ (0.1% and 0.2%) and two levels of H₃BO₄ (0.1% and 0.2%) and growth regulators including control (no growth regulator), two levels of 28-Homobrassinolide (0.5 and 1 ppm) and two levels of   Putrescine (10 and 20ppm) which were applied as foliar spray to chilli plants.  The experimental finding revealed that application of micronutrients improved pigment (chlorophyll a, chlorophyll b, total chlorophyll and carotenoids) content of leaves as well as nitrate reductase activity and relative leave water content of chilli with highest value recorded with the application of the micronutrient 0.2% Zinc (ZnSO₄) followed by 0.2% Boron (H₃BO₄) and 0.1% Zn(ZnSO₄). Foliar application of 0.2% Zinc (ZnSO₄) also recorded highest fruit yield per plant. The application of growth regulators also improved these physiological parameters with highest value recorded from the treatment 0.5 ppm of 28-Homobrassinolide followed by 1 ppm of 28-Homobrassinolide and 20 ppm Putrescine. Maximum fruit yield per plant was also recorded from 0.5 ppm of 28-Homobrassinolide. The result further revealed that fresh fruit yield per plant was positively correlated with total chlorophyll content, nitrate reductase activity and relative leaf water content.

Lentil (Lens culinaris) is an important cool season food legume, encounters numerous biotic and abiotic stresses as it contains asset of genes/proteins which helps this crop to overcome abiotic stresses and identification of stress inducible genes/proteins are an important area of research. Plant productivity is adversely affected by nature’s wrath in the form of various biotic and abiotic stresses. They cause losses worth hundreds of million dollars each year due to reduction in crop productivity and crop failure. In fact, they threaten the sustainability of agricultural industry. In the present study, primers for two abiotic stress tolerant genes namely Inositol and Alcohol dehydrogenase (ADH) were designed by Fast-PCR and validated by Primer-BLAST online tool. Stress treatment at four water potentials (-3.0, -5.0, -7.5 and -10.0 bar) was given followed by RNA isolation. Inositol and ADH were identified through PCR followed by gel elution and cloning in pGEM-T easy and pTZ57R/T vectors respectively. Confirmed clones were sequenced and obtained 1042 bp (Inositol) & 1343 bp (ADH) and analysed through bioinformatics software’s viz. BioEdit, ORF Finder, Mega5, Expasy Protparam tool. Further, biochemical studies in PEG-6000 induced stress showed increased MDA and proline content whereas decreased carbohydrate and protein content as compared to water treated control.

Grafting Malus doumeri, an apple tree, is becoming important as plantation areas are increasing in recent years. The effects of rootstock ages and scion sources on success rate, shoot number, and growth of M. doumeri were conducted in North Vietnam by using the sub-side grafting technique. The results indicated that at both three and five months of growth, the ages of rootstock significantly affected shoot number and shoot length. While at five months of growth success rate of grafting was higher in using rootstock of 2-years old. The ratio was 88.9% in 2-year-old rootstocks and that was 65.1% in the 1-year-old rootstock. At five months of growth, shoot number was 1.6, and shoot length was 55.2 cm in using 2-year-old rootstocks. Scion sources did not significantly affect the success rate at both three and five months of growth. Meanwhile, they did significantly affect shoot number and shoot length. The shoot number reduced between three and five months of growth due to competitions for light and nutrients, and ice rain. It is concluded that to graft M. doumeri trees, rootstocks of 2-years old should be used by sub-side technique. While looking for suitability between rootstock and scion source/cultivars should be taken care of for higher success rate and better growth of shoots on the scion.

The aim of this study is to conduct an ethnobotanical survey, to determine chemical composition and to evaluate antibacterial activities of essential oils of Thymus algeriensis from Imouzzer, Ammi visnaga and Myrtus communis from Taounate (Morocco). The obtained yields of Essential oils from Thymus algeriensis, Ammi visnaga and Myrtus communis were respectively 2.25%, 0.61% and 0.47%.

Essential oil of T. algeriensis was dominated by carvacrol (34.34%), δ-3-carene (20.18%), γ-terpinolene (15.09%), cis-sabinene (10.35%), germacrene D (5.15%), camphene (4.14%) and α-pinene (2.98%). Terminalol (43.35%), linalool butyrate (37.86%), limonene (5.49%), isoamyl methyl-2-butyrate (5.12%) were the major constituents of the essential oil of Ammi visnaga, while the essential oil of M. communis was dominated by δ-carene (32.9%), α-pinene (23.92%), neryl butyrate (9.95%), linalyl acetate (9.38%), α-terpineol (4.71%), methyl eugenol (4.54%), 1,8-cineole (2.41%) and α-guaiene (2.22%).

The antimicrobial activity of these essential oils was evaluated against four bacterial strains; Bacillus cereus, Staphylococcus aureus, Escherichia coli and Salmonella typhi.

Escherichia coli, Bacillus cereus and Staphylococcus aureus were inhibited from the concentration of 0.7 µl with respective inhibition zones of 1.2; 2 and 1.7 mm.

The essential oil of Thymus algeriensis showed strong inhibitory activity against Gram-positive and Gram-negative bacteria, followed by Ammi visnaga and Myrtus communis respectively. This bioactivity is mainly due to the presence of linalool and its derivatives, known for their effectiveness against microbial agents.

Phytochemicals from Curcuma amada extract is used to cure atopic dermatitis (Eczema). Atopic dermatitis is caused by Staphylococcus aureus. “Molecular docking of the phytochemicals with the enzyme was studied using “Biovia Discovery Studio”. High positive values of “-CDOCKER energy and -CDOCKER interaction energy” suggested that curcumin and ar-turmerone effectively deactivate the L - lactate dehydrogenase enzyme thereby interrupting the life cycle of the organism.

Phytochemicals from Murraya koenigii plant extract can cure the disease Eczema. Bacteria that causing Eczema is Staphylococcus aureus. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that Alpha pinene and beta-caryophyllene can deactivate the enzyme Quinone oxidoreductase that interrupting the life cycle of Staphylococcus aureus.

Phytochemicals from Beet root plant extract can cure Jaundice. It is caused by Leptospira interrogans. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that caffeice acid and myricetin can effectively deactivate the sphongomyelin phosphodiesterase enzyme thereby interrupting the life cycle of the organism.

Phytochemicals from Pandanus odorifer plant extract can cure Boils. It is caused by Staphylococcus aureus. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that Octadecanedioic acid can effectively deactivate the shikimate dehydrogenase enzyme thereby interrupting the life cycle of the organism.

Phytochemicals from Psidium guajava plant extract can cure Ulcer. It is caused by Helicobacter pylori. This objective of the study is to identify the phytochemical of Psidium guajava capable of curing Ulcer. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that p-cymene can effectively deactivate the dihydrofolate reductase enzyme thereby interrupting the life cycle of the organism.

Phytochemicals from that Celastrus paniculatus plant extract can be used to cure skin diseases. It is caused by Staphylococcus aureus. “Molecular docking method” of the phytochemicals with the enzyme was studied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that the phytochemicals alkaloids, stearic acid, betacyperonethe (benzene), and polyalcohol (ethanol) can effectively deactivate the L-lactate dehydrogenase enzyme thereby interrupting the life cycle of Staphylococcus aureus.

Phytochemicals from Kewda flower (Pandanus odorifer) extract is traditionally employed for curing ulcer. The ulcer is caused by Helicobacter pylori. Biovia Discovery Studio was used to study the molecular docking of the phytochemicals with the enzyme. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that “Heptane and 1,2,3-benzene triol” can effectively deactivate the Sikhimate dehydrogenase enzyme thereby discontinuing the life cycle of  Helicobacter pylori.

Phytochemicals from Salvia rosmarinus plant extract can cure cough. It is caused by Bordetella pertussis. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that Rosmarinic Acid can effectively deactivate the enzymes of Bordetella pertussis thereby interrupting the life cycle of the organism.

Phytochemicals from Pavonia odorate plant extract can cure Athletes Foot disease. It is caused by Epidermophyton Floccosum. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy ”suggested that palmitic acid and caproic acid can effectively deactivate the glycerol kinase epidermophyton enzyme thereby interrupting the life cycle of the organism.

Phytochemicals from Beet root plant extract can cure Hepatitis. It is caused by Hepatitis A virus. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that Myricetin can effectively deactivate the Triacylglycerol lipase enzyme thereby interrupting the life cycle of the organism.

Phytochemicals from Nigella sativa plant extract can cure Eczema. It is caused by Staphylococcus aureus. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that Carvacrol can effectively deactivate the shikimate dehydrogenase enzyme thereby interrupting the life cycle of the organism.

The phytochemicals or secondary metabolites from extract of Citrus reticulata may cure dysentery, primarily which is caused by Entamoeba hystolytica. To restrict the activity of Entamoeba hystolytica, several bio molecules can be deployed, of which the phytochemicals can be the best alternative. Molecular docking-based screening of a few phytochemicals revealed that few of phytochemicals effectively associate with the active site of the Glyceraldehydes- 3 – Phosphate and hence bears diagnostic and therapeutic potentials against Entamoeba hystolytica causing dysentry. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” indicated that the Anthranalic acid may effectively inhibit the glyceraldehydes-3-phosphate dehydrogenase enzyme thereby interrupting the life cycle of the organism.

Phytochemicals are plant derived biomolecules which are believed to protect cells from damage. It has been reported that Moringa oleifera L. (Moringaceae) is n economically important plant, distributed across tropics and subtropics whose extract is used to cure malaria. The plant extract contains different phytochemicals. Malaria is caused by Protozoan Parasite Plasmodium. L-Lactate dehydrogenase is one of its vital enzymes. In silico approach using Biovia’s Discovery Studio was used to see the molecular docking between phytochemicals derived from Moringa oleifera L. and the enzyme of interest. -CDOCKER energy and –CDOCKER interaction energy represents the strength of the interaction. Particularly, the phytochemicals hexadecanoic acid and anthraquinone can effectively deactivate the L-Lactate dehydrogenase enzyme and thus can interfere with the life cycle of Plasmodium.

Bioactive substance or Phytochemicals from Pavonia odorata plant extract can cure dermatophytosis. It is caused by Trichophyton rubrum. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that hexahydrofarnesyl acetone can effectively deactivate the laccase enzyme thereby interrupting the life cycle of Trichophyton rubrum.

The G1/01 strain is a superior variant selected from the mutation induction with Ethyl Methane Sulfonate (EMS) mutagen. This strain has a higher level of spiciness and higher capsaicin content compared to its original type. Following variations that emerge until the fourth generation of G1/01 mutant offspring, it is important to identify the appearance of mutants that have certain desired trait. In this study, morphological, physiological and molecular identification with Random Amplified Polymorphic DNA (RAPD) was conducted to identify mutants that have high spiciness and produce fruit in large quantities. Fifteen fourth generation G1/01 plants were planted. The morphology characters includes, stems, leaves, flowers and fruit were observed with refer to the Descriptors for Capsicum (Capsicum spp.) By the International Plant Genetic Resources Institute (IPGRI), Asian Vegetable Research and Development Center (AVRDC) and Tropical Agricultural Research and Training Center (CATIE) (1995). Observation of capsaicinoid content to predict the spiciness of the fruit is done by spectrophotometric methods. Furthermore, molecular distances based on Random Amplified Polymorphic DNA (RAPD) data are used to estimate the magnitude of the molecular changes occurred. Five high pungency mutants derived from G1/01 mutant lines were selected  i.e. plants no1, 10, 11, 12, and 14 based on the  high levels of capsaicinoid. Of the five plants, plant number 12 is a plant that produces fruit with high capsaicin (8.556 mg / g) so that it has a high spiciness level of up to 136.896 Schoville Units (SHU). Plants with lower spiciness (99.200 SHU) is plant no. 10, but has more fruit. Molecularly, plant no. 12 is in one cluster with several other plants so that it does not show high specificity when compared to plant no 10. Thus both mutants are superior mutant candidates with high spiciness.

Papaya production is affected by sexual variability in the seed. Conventional methods of propagation involve the maintenance of large land scale crops to supply the consumer's demand. Micropropagation techniques using temporary immersion of twin vessels (TIS) bioreactors are an alternative for papaya production. The objective of the study is to establish a disinfection protocol for the introduction of axillary buds and to evaluate different immersion times and inoculum density in the TIS to obtain a higher multiplication rate. Disinfection of the plant material was performed with 70% ethanol for 1 minute, 2% sodium hypochlorite for 5, 6 and 8 min, and sterile water washes. Percentage of contamination and oxidation was evaluated for 21 days. The introduction stage, Murashige & Skoog (MS) medium was used with 1 mg.L^-1 of gibberellic acid and 2 mg.L^-1 of kinetin. Similarly for the multiplication stage, MS medium was supplemented with 0.5 mg.L^-1 of benzyl adenine, 0.5 mg.L^-1 of indoleacetic acid, and 0.3 mg.L^-1 of gibberellic acid. The multiplication rate, number of leaves, height, and diameter of the stem was evaluated for 21 days. The plant material was transferred to the TIS to evaluate the immersion time (1 and 2 min) and density of the inoculum (4 and 8) in 200 mL of liquid multiplication medium, with an immersion frequency of 6 h, photoperiod of 12 h of lighting per day and 28°C. In the disinfection stage, 100% survival was achieved without oxidation using 2% sodium hypochlorite for 6 and 8 min. Survival rate was 96.4% and 83.33% for the introduction and multiplication stage in semi-solid medium, respectively. For the latter, 1.88 ± 0.02 leaves were observed, 1.12 ± 0.04 cm in height, diameter of 0.43 ± 0.01 cm, and a multiplication rate of 1.20 ± 0.02. In the TIS, 6.03 ± 0.04 leaves were observed, 1.65 ± 0.10 cm in height, diameter 0.73 ± 0.02, and a multiplication rate of 5.05 ± 0.06 using 2 min of immersion every 6 h with an inoculum density of eight. Research established that high multiplication was obtained by increasing immersion of plant material in liquid culture medium. This study will contribute to the strengthening of the productive sector, offering seedlings with the suitable sex (hermaphrodite), and high genetic and phytosanitary quality.

The screening of new components from medicinal plants needs several screenings. In this review, we report our results obtained with the use of medicinal plants of the North-West of Morocco and their biological properties. The region possesses a distinguish climate and floristic diversity, including medicinal plants that have been used to treat several diseases. The secondary metabolites of these plants possess important charges of phenolic, flavonoid and terpenoid compounds. Moreover, these medicinal plants showed remarkable biological properties such as antibacterial, antiparasitic, antioxidant and anticancer effects. The findings suggest that Medicinal plants of the North-West constitute a veritable source for drugs development.