RESUMEN
The possible involvement of polyamines in the chilling tolerance of spinach (Spinacia oleracea L.) was investigated focusing on photosynthesis. During chilling at 8/5C (day/night) for 6 d, S-adenosylmethionine decarboxylase (SAMDC) activity increased significantly in leaves in parallel with the increase in putrescine and spermidine (Spd) content in leaves and chloroplasts. Treatment of leaves with methylglyoxal-bis(guanylhydrazone) (MGBG), an SAMDC inhibitor, resulted in the deterioration of plant growth and photosynthesis under chilling conditions, which was reversed by the concomitant treatment with Spd through the roots. Plants treated with MGBG showed lower photochemical efficiency of PSII than either the control or plants treated with MGBG plus Spd during chilling and even after transfer to warm conditions, suggesting an increase of photoinhibition due to low Spd in chloroplasts. Indeed, MGBG-treated plants had much lower activities of thylakoid electron transport and enzymes in carbon metabolism as well as higher degrees of lipid peroxidation of thylakoid membranes compared to the control. These results indicate that the enhanced activity of SAMDC with a consequential rise of Spd in chloroplasts is crucial for the cold acclimation of the photosynthetic apparatus in spinach leaves.
Asunto(s)
Aclimatación/fisiología , Adenosilmetionina Descarboxilasa/metabolismo , Fotosíntesis/fisiología , Hojas de la Planta/enzimología , Spinacia oleracea/enzimología , Adenosilmetionina Descarboxilasa/genética , Carboxiliasas/genética , Carboxiliasas/metabolismo , Clorofila/metabolismo , Clorofila/efectos de la radiación , Cloroplastos/metabolismo , Frío , Transporte de Electrón/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Luz , Complejos de Proteína Captadores de Luz , Peroxidación de Lípido/efectos de los fármacos , Mitoguazona/farmacología , Fotosíntesis/efectos de la radiación , Proteínas del Complejo del Centro de Reacción Fotosintética/efectos de los fármacos , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/metabolismo , Poliaminas/metabolismo , Putrescina/metabolismo , ARN Mensajero/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Espermidina/metabolismo , Spinacia oleracea/crecimiento & desarrollo , Tilacoides/efectos de los fármacos , Tilacoides/metabolismo , Tilacoides/efectos de la radiaciónRESUMEN
Polyamines play pivotal roles in plant defense to environmental stresses. However, stress tolerance of genetically engineered plants for polyamine biosynthesis has been little examined so far. We cloned spermidine synthase cDNA from Cucurbita ficifolia and the gene was introduced to Arabidopsis thaliana under the control of the cauliflower mosaic virus 35S promoter. The transgene was stably integrated and actively transcribed in the transgenic plants. As compared with the wild-type plants, the T2 and T3 transgenic plants exhibited a significant increase in spermidine synthase activity and spermidine content in leaves together with enhanced tolerance to various stresses including chilling, freezing, salinity, hyperosmosis, drought, and paraquat toxicity. During exposure to chilling stress (5 degrees C), the transgenics displayed a remarkable increase in arginine decarboxylase activity and conjugated spermidine contents in leaves compared to the wild type. A cDNA microarray analysis revealed that several genes were more abundantly transcribed in the transgenics than in the wild type under chilling stress. These genes included those for stress-responsive transcription factors such as DREB and stress-protective proteins like rd29A. These results strongly suggest an important role for spermidine as a signaling regulator in stress signaling pathways, leading to build-up of stress tolerance mechanisms in plants under stress conditions.