RESUMEN
Thermal stress induces a wide array of morphological and physiological changes in potato affecting its development and economic yield. Response to thermal stress in plants is mostly regulated by heat shock factors (hsfs). The current study aimed at improving heat tolerance by transforming potato plant with heat shock factor, HsfA1d, using Agrobacterium. Gateway cloning strategy was adopted for isolation of HsfA1d from Arabidopsis thaliana and cloning into plant expression vector. The target gene was introduced into potato by infecting internodal explants with Agrobacterium strain GV3101 carrying pGWB402Ω-HsfA1d construct. Upon exposure to heat stress, the wild-type plants turned yellowish, whereas no phenotypic effect on transgenic plants was observed. Expression of HsfA1d in transgenic plants was increased by 5.8-fold under thermal stress compared to room temperature. Transgenic plants exhibited 6-fold increase in the expression of downstream HSP70 under thermal stress compared to wild-type plants. Both chlorophyll a and b were significantly decreased in wild-type plants while no such decrease was recorded in transgenic plants under thermal stress. Heat stress was found to have no significant effect on carotenoid pigments of both wild-type and transgenic plants. Significantly lower electrolyte leakage from transgenic plants was witnessed compared to wild type upon exposure to thermal stress. Transgenic plants accumulated significantly higher proline content compared to wild-type plants under heat stress. It is concluded that HsfA1d plays a vital role in plant thermotolerance and hence can be effectively used to enhance the resistance of crop plants against heat stress.
Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas/genética , Factores de Transcripción del Choque Térmico/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Genes de Plantas , Factores de Transcripción del Choque Térmico/metabolismo , Respuesta al Choque Térmico/genética , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/genética , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Termotolerancia/genética , Termotolerancia/fisiología , Factores de Transcripción/metabolismoRESUMEN
Medicinal and aromatic plants contribute to major portion of the flora. The plant materials obtained from these plants are used in the pharmaceuticals, cosmetics, and drug industries. Tamarix dioica is locally used in the management of splenic and hepatic inflammation as well as diuretic and carminative. It also possesses cytotoxic, antimicrobial, antifungal and anti-inflammatory activity. The present study investigates the anti-inflammatory, anti-pyretic and analgesic activities of the crude methanolic extract from Tamarix dioica. Anti-inflammatory activity was measured by Carrageenan Induced Paw Edema and Xylene Induced Ear Edema methods. Pyrexia induction with Brewer's yeast assay was used to determine antipyretic activity and analgesic activity was estimated by acetic acid induced writhing and hot plate methods. The data indicated that anti-inflammatory, anti-pyretic and analgesic activities of the crude methanolic extract from Tamarix dioica was dose and time dependent when measured by different assays. Exposure of model animal to increasing concentrations of the plant extract for longer period increased their anti-inflammatory, anti-pyretic and analgesic activities. Significantly highest anti-inflammatory, anti-pyretic and analgesic activities were noted at highest doses of the crude methanolic extract for longer exposure compared with their respective controls.
Asunto(s)
Analgésicos/farmacología , Antiinflamatorios/farmacología , Antipiréticos/farmacología , Tamaricaceae/química , Ácido Acético/farmacología , Analgésicos/química , Animales , Antiinfecciosos/química , Antiinfecciosos/farmacología , Antiinflamatorios/química , Antifúngicos/química , Antifúngicos/farmacología , Antipiréticos/química , Modelos Animales de Enfermedad , Edema/tratamiento farmacológico , Edema/microbiología , Femenino , Inflamación/tratamiento farmacológico , Hígado/efectos de los fármacos , Masculino , Metanol/química , Fitoterapia/métodos , Extractos Vegetales/química , Extractos Vegetales/farmacología , Bazo/efectos de los fármacosRESUMEN
A high molecular weight serine protease has been purified to electrophoretic homogeneity from the seeds of Caesalpinia bonducella Flem. (Caesalpiniaceae) by the combination of size exclusion and ion exchange chromatography. About 524 fold purification was achieved with an overall recovery of 6.8%. The specific activity was found to be 86 U/mg/min at pH 8.0. The calculated K(m) and V(max) were 1.66 mg/mL and 496.68 units/min per mg of protein, respectively. The molecular mass was estimated to be about 63 kDa by sodium dodecyl sulfate PAGE. The enzyme showed optimum activity at pH 8.0 and exhibited its highest activity at 40 degrees C. The enzyme was strongly inhibited by 2mM phenylmethylsulfonyl fluoride (PMSF), suggesting the presence of a serine residue at the active site. PMSF showed a pure competitive type of inhibition with the serine protease enzyme. It was observed that enzyme activity was enhanced in the presence of dications and was active against a variety of modified substrates and natural proteins.