RESUMEN
Chilling causes a significant decline in photosynthesis in tomato plants. Tomato tryptophan decarboxylase gene 1 (SlTDC1) is the first rate-limiting gene for melatonin (MT) biosynthesis and is involved in the regulation of photosynthesis under various abiotic stresses. However, it is not clear whether SlTDC1 participates in the photosynthesis of tomato under chilling stress. Here, we obtained SlTDC1 overexpression transgenic tomato seedlings, which showed higher SlTDC1 mRNA abundance and MT content compared with the wild type (WT). The results showed that the overexpression of SlTDC1 obviously alleviated the chilling damage to seedlings in terms of the lower electrolyte leakage rate and hydrogen peroxide content, compared with the WT after 2 d of chilling stress. Moreover, the overexpression of SlTDC1 notably increased photosynthesis under chilling stress, which was related to the higher chlorophyll content, normal chloroplast structure, and higher mRNA abundance and protein level of Rubisco and RCA, as well as the higher carbon metabolic capacity, compared to the WT. In addition, we found that SlTDC1-overexpressing seedlings showed higher Wk (damage degree of OEC on the PSII donor side), φEo (quantum yield for electron transport in the PSII reaction center), and PIABS (photosynthetic performance index) than WT seedlings after low-temperature stress, implying that the overexpression of SlTDC1 decreased the damage to the reaction center and donor-side and receptor-side electron transport of PSII and promoted PSI activity, as well as energy absorption and distribution, to relieve the photoinhibition induced by chilling stress. Our results support the notion that SlTDC1 plays a vital role in the regulation of photosynthesis under chilling stress.
Asunto(s)
Solanum lycopersicum , Solanum lycopersicum/genética , Dióxido de Carbono , Plantas Modificadas Genéticamente/metabolismo , Fotosíntesis/genética , Frío , ARN Mensajero/metabolismoRESUMEN
KEY MESSAGE: Rootstock provides more abscisic acid (ABA) content to scions to increase the chilling tolerance of seedlings. H2O2 is involved in ABA regulation of grafting-induced chilling tolerance of cucumber. Here we examined the role of ABA in the response of grafted cucumber to chilling stress. The data showed chilling induced an increase in leaf and root ABA content and there was a positive correlation between ABA content and the chilling tolerance of the varieties. The increase of ABA content and NCED mRNA abundance in the leaf of both Cs/Cs (self-root) and Cs/Cm (grafted with pumpkin as rootstock) showed a delay under aerial stress compared with those under whole plant and root-zone stress. Intriguingly, an increase in ABA in xylem was found under whole-plant and root-zone chilling stress but was not detected under aerial stress, implying the increases in ABA content in leaves were mainly from root ABA transportation. Compared to Cs/Cs, a higher ABA content and NCED mRNA abundance were observed in Cs/Cm, which showed that Cm could output more ABA than Cs. The removal of endogenous ABA decreased the difference in chilling tolerance induced by Cm, as evidenced by the observed similar oxidative stress levels and photosynthetic capacity between Cs/Cs and Cs/Cm after chilling stress. Moreover, we found that the H2O2 signal in grafted cucumber could respond to chilling stress earlier than the H2O2 signal in self-rooted cucumber. The inhibition of endogenous H2O2 decreased the chilling tolerance of grafted cucumber induced by ABA by reducing photosynthesis and the mRNA abundance of CBF1 and COR. Thus, our results indicate that H2O2, as the downstream signal, participated in the rootstock-induced chilling tolerance of grafted seedlings induced by ABA.
Asunto(s)
Cucumis sativus , Ácido Abscísico , Cucumis sativus/genética , Peróxido de Hidrógeno , Raíces de Plantas , ARN Mensajero/genética , Plantones/genéticaRESUMEN
Tomato (Solanum lycopersicum) is one of the most important greenhouse vegetables, with a large cultivated area across the world. However, in northern China, tomato plants often suffer from low-temperature stress in solar greenhouse cultivation, which affects plant growth and development and results in economic losses. We previously found that a chloroplast aldolase gene in tomato, SlFBA4, plays an important role in the Calvin-Benson cycle (CBC), and its expression level and activity can be significantly altered when subjected to low-temperature stress. To further study the function of SlFBA4 in the photosynthesis and chilling tolerance of tomato, we obtained transgenic tomato plants by the over-expression and RNA interference (RNAi) of SlFBA4. The over-expression of SlFBA4 led to higher fructose-1,6-bisphosphate aldolase activity, net photosynthetic rate (Pn) and activity of other enzymes in the CBC than wild type. Opposite results were observed in the RNAi lines. Moreover, an increase in thousand-seed weight, plant height, stem diameter and germination rate in optimal and sub-optimal temperatures was observed in the over-expression lines, while opposite effects were observed in the RNAi lines. Furthermore, over-expression of SlFBA4 increased Pn and enzyme activity and decreased malonaldehyde (MDA) content under chilling conditions. On the other hand, Pn and MDA content were more severely influenced by chilling stress in the RNAi lines. These results indicate that SlFBA4 plays an important role in tomato growth and tolerance to chilling stress.
Asunto(s)
Adaptación Fisiológica , Cloroplastos/genética , Cloroplastos/metabolismo , Frío , Serpinas/genética , Solanum lycopersicum/fisiología , Biomarcadores , Regulación de la Expresión Génica de las Plantas , Fenotipo , Fotosíntesis/genética , Proteínas de Plantas/genética , Plantas Modificadas GenéticamenteRESUMEN
Melatonin (MT), as a signaling molecule, plays a vital role in regulating leaf senescence in plants. This study aimed to verify the antioxidant roles of MT in delaying dark- or age-induced leaf senescence of cucumber plants. The results showed that endogenous MT responds to darkness and overexpression of CsASMT, the key gene of MT synthesis, and delays leaf senescence stimulated by darkness, as manifested by significantly lower malonaldehyde (MDA) and reactive oxygen species (ROS) contents as well as higher activities and gene expression of antioxidant enzymes compared to the control. Moreover, MT suppressed both age- or dark-induced leaf senescence of cucumber, as evidenced by a decrease in senescence-related gene SAG20 and cell-death-related gene PDCD expression and ROS content and an increase in antioxidant capacity and chlorophyll biosynthesis compared with the H2O-treated seedlings. Meanwhile, the suppression of age-induced leaf senescence by melatonin was also reflected by the reduction in abscisic acid (ABA) biosynthesis and signaling pathways as well as the promotion of auxin (IAA) biosynthesis and signaling pathways in cucumber plants in the solar greenhouse. Combining the results of the two separate experiments, we demonstrated that MT acts as a powerful antioxidant to alleviate leaf senescence by activating the antioxidant system and IAA synthesis and signaling while inhibiting ABA synthesis and signaling in cucumber plants.
Asunto(s)
Cucumis sativus , Melatonina , Ácido Abscísico/metabolismo , Antioxidantes/metabolismo , Cucumis sativus/genética , Cucumis sativus/metabolismo , Regulación de la Expresión Génica de las Plantas , Ácidos Indolacéticos/metabolismo , Melatonina/metabolismo , Hojas de la Planta/metabolismo , Senescencia de la Planta , Plantas/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Grafting is widely applied to enhance the tolerance of some vegetables to biotic and abiotic stress. Salicylic acid (SA) is known to be involved in grafting-induced chilling tolerance in cucumber. Here, we revealed that grafting with pumpkin (Cucurbita moschata, Cm) as a rootstock improved chilling tolerance and increased the accumulation of SA, abscisic acid (ABA) and hydrogen peroxide (H2O2) in grafted cucumber (Cucumis sativus/Cucurbita moschata, Cs/Cm) leaves. Exogenous SA improved the chilling tolerance and increased the accumulation of ABA and H2O2 and the mRNA abundances of CBF1, COR47, NCED, and RBOH1. However, 2-aminoindan-2-phosphonic acid (AIP) and L-a-aminooxy-b-phenylpropionic acid (AOPP) (biosynthesis inhibitors of SA) reduced grafting-induced chilling tolerance, as well as the synthesis of ABA and H2O2, in cucumber leaves. ABA significantly increased endogenous H2O2 production and the resistance to chilling stress, as proven by the lower electrolyte leakage (EL) and chilling injury index (CI). However, application of the ABA biosynthesis inhibitors sodium tungstate (Na2WO4) and fluridone (Flu) abolished grafting or SA-induced H2O2 accumulation and chilling tolerance. SA-induced plant response to chilling stress was also eliminated by N,N'-dimethylthiourea (DMTU, an H2O2 scavenger). In addition, ABA-induced chilling tolerance was attenuated by DMTU and diphenyleneiodonium (DPI, an H2O2 inhibitor) chloride, but AIP and AOPP had little effect on the ABA-induced mitigation of chilling stress. Na2WO4 and Flu diminished grafting- or SA-induced H2O2 biosynthesis, but DMTU and DPI did not affect ABA production induced by SA under chilling stress. These results suggest that SA participated in grafting-induced chilling tolerance by stimulating the biosynthesis of ABA and H2O2. H2O2, as a downstream signaler of ABA, mediates SA-induced chilling tolerance in grafted cucumber plants.
Asunto(s)
Cucumis sativus , Ácido Abscísico/farmacología , Peróxido de Hidrógeno/farmacología , Ácido Salicílico/farmacología , Productos Avanzados de Oxidación de Proteínas/farmacologíaRESUMEN
As an important gas signaling molecule, hydrogen sulfide (H2S) plays a crucial role in regulating cold tolerance. H2S cooperates with phytohormones such as abscisic acid, ethylene, and salicylic acid to regulate the plant stress response. However, the synergistic regulation of H2S and auxin in the plant response to cold stress has not been reported. This study showed that sodium hydrosulfide (NaHS, an H2S donor) treatment enhanced the cold stress tolerance of cucumber seedlings and increased the level of auxin. CsARF5, a cucumber auxin response factor (ARF) gene, was isolated, and its role in regulating H2S-mediated cold stress tolerance was described. Transgenic cucumber leaves overexpressing CsARF5 were obtained. Physiological analysis indicated that overexpression of CsARF5 enhanced the cold stress tolerance of cucumber and the regulation of the cold stress response by CsARF5 depends on H2S. In addition, molecular assays showed that CsARF5 modulated cold stress response by directly activating the expression of the dehydration-responsive element-binding (DREB)/C-repeat binding factor (CBF) gene CsDREB3, which was identified as a positive regulator of cold stress. Taken together, the above results suggest that CsARF5 plays an important role in H2S-mediated cold stress in cucumber. These results shed light on the molecular mechanism by which H2S regulates cold stress response by mediating auxin signaling; this will provide insights for further studies on the molecular mechanism by which H2S regulates cold stress. The aim of this study was to explore the molecular mechanism of H2S regulating cold tolerance of cucumber seedlings and provide a theoretical basis for the further study of cucumber cultivation and environmental adaptability technology in winter.
Asunto(s)
Cucumis sativus/crecimiento & desarrollo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Sulfuros/farmacología , Respuesta al Choque por Frío/efectos de los fármacos , Cucumis sativus/efectos de los fármacos , Cucumis sativus/genética , Cucumis sativus/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Sulfuro de Hidrógeno/metabolismo , Ácidos Indolacéticos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/efectos de los fármacos , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Plantas Modificadas Genéticamente/metabolismoRESUMEN
Hydrogen sulfide (H2S) plays a crucial role in regulating chilling tolerance. However, the role of hydrogen peroxide (H2O2) and auxin in H2S-induced signal transduction in the chilling stress response of plants was unclear. In this study, 1.0 mM exogenous H2O2 and 75 µM indole-3-acetic acid (IAA) significantly improved the chilling tolerance of cucumber seedlings, as demonstrated by the mild plant chilling injury symptoms, lower chilling injury index (CI), electrolyte leakage (EL), and malondialdehyde content (MDA) as well as higher levels of photosynthesis and cold-responsive genes under chilling stress. IAA-induced chilling tolerance was weakened by N, N'-dimethylthiourea (DMTU, a scavenger of H2O2), but the polar transport inhibitor of IAA (1-naphthylphthalamic acid, NPA) did not affect H2O2-induced mitigation of chilling stress. IAA significantly enhanced endogenous H2O2 synthesis, but H2O2 had minimal effects on endogenous IAA content in cucumber seedlings. In addition, the H2O2 scavenger DMTU, inhibitor of H2O2 synthesis (diphenyleneiodonium chloride, DPI), and IAA polar transport inhibitor NPA reduced H2S-induced chilling tolerance. Sodium hydrosulfide (NaHS) increased H2O2 and IAA levels, flavin monooxygenase (FMO) activity, and respiratory burst oxidase homolog (RBOH1) and FMO-like protein (YUCCA2) mRNA levels in cucumber seedlings. DMTU, DPI, and NPA diminished NaHS-induced H2O2 production, but DMTU and DPI did not affect IAA levels induced by NaHS during chilling stress. Taken together, the present data indicate that H2O2 as a downstream signal of IAA mediates H2S-induced chilling tolerance in cucumber seedlings.
Asunto(s)
Aclimatación , Cucumis sativus/metabolismo , Peróxido de Hidrógeno/metabolismo , Sulfuro de Hidrógeno/metabolismo , Ácidos Indolacéticos/metabolismo , Frío , Plantones/metabolismoRESUMEN
MAIN CONCLUSION: This report proves a cross talk between H2S and IAA in cold stress response, which has presented strong evidence that IAA acts as a downstream signal mediating the H2S-induced stress tolerance in cucumber seedlings. We evaluated changes in endogenous hydrogen sulfide (H2S) and indole-3-acetic acid (IAA) emission systems, and the interactive effect of exogenous H2S and IAA on chilling tolerance in cucumber seedlings. The results showed that chilling stress increased the activity and relative mRNA expression of L-/D-cysteine desulfhydrase (L-/D-CD), which in turn induced the accumulation of endogenous H2S. Similarly, the endogenous IAA system was triggered by chilling stress. We found that 1.0 mM sodium hydrosulfide (NaHS, an H2S donor) significantly enhanced the activity of flavin monooxygenase (FMO) and relative expression of FMO-like proteins (YUCCA2), which in turn elevated endogenous IAA levels in cucumber seedlings. However, IAA had little effects on activities of L-/D-CD and endogenous H2S levels. H2S-induced IAA production accompanied by increase in chilling tolerance, as shown by the decrease in stress-induced electrolyte leakage (EL) and reactive oxygen species (ROS) accumulation, and increase in gene expressions and enzyme activities of photosynthesis. 1-naphthylphthalamic acid (NPA, an IAA polar transport inhibitor) declined H2S-induced chilling tolerance and defense genes' expression. However, scavenging of H2S had a little effect on IAA-induced chilling tolerance. These results suggest that IAA acting as a downstream signaling molecule is involved in the H2S-induced chilling tolerance in cucumber seedlings.
Asunto(s)
Aclimatación/efectos de los fármacos , Cucumis sativus/fisiología , Sulfuro de Hidrógeno/farmacología , Ácidos Indolacéticos/farmacología , Transducción de Señal/efectos de los fármacos , Aclimatación/genética , Aclimatación/fisiología , Ácido Ascórbico/análisis , Frío , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Glutatión/análisis , Especies Reactivas de Oxígeno/metabolismo , Plantones/genética , Plantones/fisiología , Estrés Fisiológico/efectos de los fármacos , Sulfuros , TranscriptomaRESUMEN
Despite being a key enzyme of Cavin cycle, transketolase (TK) is believed to be related to abiotic resistance in higher plants. However, how TK affects chilling tolerance still remains largely unknown. Here, we describe the effect of overexpression of the Cucumis sativa TK gene (CsTK) on growth, photosynthesis, ROS metabolism and cell ultrastructure under chilling stress. Low temperature led to a decrease of the photosynthetic rate (Pn), the stomatal conductance (Gs), the actual photochemical efficiency (ΦPSII) and the sucrose content, whereas there was an increase of the intercellular CO2 concentration (Ci) and MDA content. These changes were alleviated in the CsTK plants after 5 days of chilling stress, however, inhibition of CsTK showed the opposite results. Furthermore, transgenic plants with overexpression of CsTK showed higher increase in leaf area and dry matter, higher activity of the enzymes and higher increase in the contents of metabolism substance involved in Calvin cycle and reactive oxygen scavenging system as well as lower ⢠OH and H2 O2 content, superoxide anion production rate compared with the control cucumber plants under chilling stress. At the end of the chilling stress, compared to wild-type (WT) which exhibited dramatically destroyed cell ultrastructure, expanded chloroplast, broken cell and chloroplast membranes as well as the disappeared grana lamella, the CsTK sense plants showed a more complete cell ultrastructure, whereas, the damage of the cell ultrastructure was aggravated in CsTK antisense plants. Taken together, these results imply that CsTK promoted chilling tolerance in cucumber plants mainly through increasing the capacity to assimilate carbon, alleviating oxidative damage and stabilizing cell structure.
Asunto(s)
Frío , Cucumis sativus/enzimología , Estrés Oxidativo , Fotosíntesis , Transcetolasa/genética , Cucumis sativus/genética , Proteínas de Plantas/genéticaRESUMEN
Elevated atmospheric CO2 alleviates moderate to severe drought stresses at physiological level in cucumber. To investigate the underlying metabolic mechanisms, cucumber seedlings were treated with two [CO2] and three water treatments combinations, and their leaves were analyzed using a non-targeted metabolomics approach. The results showed that elevated [CO2] changed 79 differential metabolites which were mainly associated with alanine, aspartate and glutamate metabolism; arginine and proline metabolism; TCA cycle; and glycerophospholipid metabolism under moderate drought stress. Moreover, elevated [CO2] promoted the accumulation of secondary metabolites; including isoferulic acid, m-coumaric acid and salicyluric acid. Under severe drought stress, elevated [CO2] changed 26 differential metabolites which mainly involved in alanine, aspartate and glutamate metabolism; pyruvate metabolism; arginine and proline metabolism; glyoxylate and dicarboxylate metabolism; cysteine and methionine metabolism; starch and sucrose metabolism; glycolysis or gluconeogenesis; and pyrimidine metabolism. In addition, elevated [CO2] accumulated carbohydrates, 1,2,3-trihydroxybenzene, pyrocatechol, glutamate, and l-gulonolactone, to allow adaption to severe drought. In conclusion, the metabolites and metabolic pathways associated with the alleviation of drought stresses by elevated [CO2] were different according to the level of drought stress. Our results may provide a theoretical basis for CO2 fertilization and application of exogenous metabolites to enhance drought tolerance of cucumber.
Asunto(s)
Presión Atmosférica , Dióxido de Carbono/metabolismo , Cucumis sativus/metabolismo , Sequías , Metabolómica , Hojas de la Planta/metabolismo , Plantones/metabolismo , Dióxido de Carbono/análisisRESUMEN
In northern China, low temperature is the most common abiotic stresses for tomato plants cultivated in solar-greenhouse in winter. We recently found that the expression and enzyme activity of fructose-1,6-bisphosphate aldolases (FBAs) in tomato, which are important enzymes in the Calvin-Benson cycle (CBC), were significantly altered in tomato seedlings subjected to heat/cold stresses. In order to study the role of FBA in photosynthesis and in regulating cold stress responses of tomato seedlings (Solanum lycopersicum), we transformed a tomato inbred line (FF) with RNA interference (RNAi) vector containing SlFBA7 reverse tandem repeat sequence. We found that the decreased SlFBA7 expression led to the decreased activities of FBA, as well as the activities of other main enzymes in the CBC. We also noticed a decrease in net photosynthetic rate, ribulose-1,5-bisphosphate and soluble sugar content, stem diameter, dry weight and seed size in RNAi SlFBA7 plants compared to wild-type. However, there are no changes in starch contents in the RNAi transgenic plants. RNAi SlFBA7 plants showed a decreased germination rate, and an increased levels of superoxide anions (O2·- ) and hydrogen peroxide (H2 O2 ) under low temperature (8/5°C) and low-light intensity (100 µmol m-2 s-1 photon flux density) growth conditions. These findings demonstrated the important role of SlFBA7 in regulating growth and chilling tolerance of tomato seedlings, and suggested that the catalytic activity of FBA in the CBC is sensitive to temperature.
Asunto(s)
Aldehído-Liasas/metabolismo , Solanum lycopersicum/enzimología , Aldehído-Liasas/genética , Frío , Regulación de la Expresión Génica de las Plantas , Solanum lycopersicum/genética , Solanum lycopersicum/crecimiento & desarrollo , Solanum lycopersicum/fisiología , Fotosíntesis , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Interferencia de ARN , Plantones/enzimología , Plantones/genética , Plantones/crecimiento & desarrollo , Plantones/fisiología , Semillas/enzimología , Semillas/genética , Semillas/crecimiento & desarrollo , Semillas/fisiología , Estrés FisiológicoRESUMEN
Rubisco activase (RCA) is an important enzyme that can catalyze the carboxylation and oxygenation activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), which is involved in the photosynthetic carbon reduction cycle. Here, we studied the effects of changes in RCA activity on photosynthesis, growth and development, as well as the low temperature and weak light tolerance of RCA overexpressing transgenic cucumber (Cucumis sativus) plants. CsRCA overexpression increased the plant height, leaf area and dry matter, and decreased the root/top ratio in transgenic cucumber plants compared with the wild-type (WT) plants. Low temperature and low light stress led to decreases in the CsRCA expression and protein levels, the photosynthetic rate (Pn) and the stomatal conductance (Gs), but an increase in the intercellular CO2 (Ci) concentration in cucumber leaves. The actual photochemical efficiency and maximal photochemical efficiency of photosystem II in cucumber seedlings also declined, but the initial fluorescence increased during low temperature and weak light stress. Transgenic plants showed a lower decrease in the CsRCA expression level and actual and maximal photochemical efficiencies, as well as increases in the Ci and initial fluorescence relative to the WT plants. Low temperature and low light stress resulted in a significant increase in the malondialdehyde (MDA) content; however, this increase was reduced in transgenic plants compared with that in WT plants. Thus, the overexpression of CsRCA may promote the growth and low temperature and low light tolerance of cucumber plants in solar greenhouses.
Asunto(s)
Aclimatación , Cucumis sativus/enzimología , Fotosíntesis , Proteínas de Plantas/metabolismo , Metabolismo de los Hidratos de Carbono , Frío , Cucumis sativus/genética , Cucumis sativus/crecimiento & desarrollo , Cucumis sativus/efectos de la radiación , Luz , Hojas de la Planta/enzimología , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Ribulosa-Bifosfato Carboxilasa/metabolismoRESUMEN
KEY MESSAGE: Four CsTK antisense transgenic cucumber plants were obtained. Decreased TK activity decreased the photosynthetic rate, seed germination rate, growth yield, and the tolerance to low temperature and weak light stress. Transketolase (TK, EC 2.2.1.1) is a key enzyme in the photosynthetic carbon reduction cycle (Calvin cycle). A cDNA fragment (526 bp) encoding transketolase was cloned from cucumber plants (Cucumis sativa L. cv 'Jinyou 3') by RT-PCR. The antisense expression [(PBI-CsTK(-)] vector containing the CsTK gene fragment was constructed. The resulting plasmid was introduced into the cucumber inbred lines '08-1' using the agrobacterium-mediated method, and four antisense transgenic cucumber plants were obtained. Decreased CsTK expression either unaltered or slightly increased the mRNA abundance and activities of the other main enzymes in the Calvin cycle, however, it decreased the TK activity and net photosynthetic rate (Pn) in antisense transgenic cucumber leaves. Antisense plants showed decreases in the growth, ratio of female flowers and yield compared with the wild-type (WT) plants. The decrease in Pn, stomatal conductance (Gs), transpiration rate (Tr), photochemical efficiency (Fv/Fm) and actual photochemical efficiency of PSII (ΦPSII) and the increase in electrolyte leakage (EL) were greater in antisense transgenic plants than in WT plants under low temperature (5 °C) and low light intensity (100 µmol m(-2) s(-1)).
Asunto(s)
Cucumis sativus/fisiología , Transcetolasa/metabolismo , Biomasa , Carbohidratos/análisis , Frío , Cucumis sativus/enzimología , Cucumis sativus/genética , Cucumis sativus/efectos de la radiación , Flores/enzimología , Flores/genética , Flores/fisiología , Flores/efectos de la radiación , Luz , Fotosíntesis , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Hojas de la Planta/efectos de la radiación , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Plantones/enzimología , Plantones/genética , Plantones/fisiología , Plantones/efectos de la radiación , Almidón/análisis , Estrés Fisiológico , Transcetolasa/genéticaRESUMEN
Grafting with pumpkin as rootstock could improve chilling tolerance of cucumber; however, the underlying mechanism of grafting-induced chilling tolerance remains unclear. Here, we analyzed the difference of physiological and transcriptional level between own-rooted (Cs/Cs) and hetero-grafted (Cs/Cm) cucumber seedlings under chilling stress. The results showed that grafting with pumpkin significantly alleviated the chilling injury as evidenced by slightly symptoms, lower contents of electrolyte leakage (EL), malondialdehyde (MDA), hydrogen peroxide (H2O2), and superoxide anion (O2-) and higher relative water content in Cs/Cm seedlings compared with Cs/Cs seedlings under chilling stress. RNA-seq data showed that grafting induced more DGEs at 8 °C/5 °C compared with 25 °C/18 °C. In accordance with the increase of the activities of antioxidant enzymes (SOD, POD, CAT, APX), grafting upregulated the expression of the regulated redox-related genes such as GST, SOD, and APX. Moreover, grafting increased the expression of genes participated in central carbon metabolism to promote the conversion and decomposition of sugar, which provided more energy for the growth of Cs/Cm seedlings under chilling stress. In addition, grafting regulated the genes involved in the intracellular signal transduction pathways such as calcium signal (CAML, CML, and CDPK) and inositol phospholipid signal (PLC), as well as changed the gene expression of plant hormone signal transduction pathways (ARF, GAI, ABF, and PYR/PYL). These results provide a physiological and transcriptional basis for the molecular mechanism of grafting-induced chilling tolerance of cucumber seedlings.
Asunto(s)
Cucumis sativus , Cucumis sativus/genética , Cucumis sativus/metabolismo , Peróxido de Hidrógeno/metabolismo , Estrés Fisiológico/genética , Perfilación de la Expresión Génica , Superóxido Dismutasa/metabolismo , Plantones/metabolismoRESUMEN
Aiming at solving the problems of soil environment deterioration and the decline of both yield and quality caused by excessive application of chemical fertilizer, we investigated the effects of rotted corn straw on the soil environment of root zone, yield and quality of cucumber with 'Jinyou 35' cucumber as the experimental material. There were three treatments, namely, combined application of rotted corn straw and chemical fertilizer (T1, the total nitrogen fertilizer application were 450 kg N·hm-2, of which 9000 kg·hm-2 rotted corn straw was used as the subsoil fertilizer, and the rest was supplemented with chemical fertilizer), pure chemical fertilizer (T2, the total nitrogen fertilizer application was the same as T1) and no fertilization (control). The results showed that the content of soil organic matter in root zone soil in T1 treatment was much higher, but no difference between T2 treatment and the control, after two continuous plantings in one year. The concentrations of soil alkaline nitrogen, available phosphorus, available potassium of T1 and T2 in cucumber root zone were higher than that in the control. T1 treatment had lower bulk density, but markedly higher porosity and respiratory rate than T2 treatment and the control in root zone soil. The electric conductivity of T1 treatment was higher than that of the control, but significantly lower than T2 treatment. There was no significant difference in pH among the three treatments. The quantity of bacteria and actinomycetes in cucumber rhizosphere soil were the highest in T1, and the lowest in the control. However, the highest quantity of fungi was found in T2. The enzyme activities of rhizosphere soil in T1 treatment were markedly higher than those of the control, whereas those of T2 treatment were significantly lower or had no significant difference relative to the control. The cucumber root dry weight and root activity of T1 were significantly higher than that of the control. The yield of T1 treatment increased by 10.1%, and fruit quality improved obviously. The root activity of T2 treatment was also significantly higher than that in the control. There was no significant difference in root dry weight and yield between T2 treatment and the control. Furthermore, T2 treatment revealed a decrease in fruit quality relative to T1 treatment. These results suggested that the combined application of rotted corn straw and chemical fertilizer could improve soil environment, promote root growth, enhance root activity and improve yield and quality of cucumber in solar-greenhouse, which could be popularized and applied in protected cucumber production.
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Cucumis sativus , Suelo , Suelo/química , Agricultura/métodos , Zea mays , Fertilizantes , Nitrógeno/análisisRESUMEN
Fulvic acid (FA) participates in the regulation of drought stress tolerance in plants, but the underlying mechanisms remain unclear. We carried out an experiment with cucumber cultivar 'Jinyou 35' as the test material and the polyethylene glycol (PEG-6000) being used to simulate drought stress. The concentration effect of FA on drought alleviation of cucumber as well as the effect of FA on photosynthetic enzymes activities, chloroplast ultrastructure, fluorescence parameters, water use efficiency, yield and quality of cucumber plants were studied through spraying FA with different concentrations (0, 100, 300, 500, 700 and 900 mg·L-1). The results showed that FA with different concentrations significantly promoted the relative water content and leaf area and decreased drought injure index as well as the electrolyte leakage and malondialdehyde (MDA) content, compared with the control (0 mg·L-1) under drought stress. The mitigative effect of FA increased first and then decreased with the increases of FA concentration, with 700 mg·L-1 FA showing the best effect. FA significantly enhanced the chlorophyll content, Rubiasco and Rubisco activase (RCA) activities and gene expression, net photosynthesis (Pn), maximal photochemical efficiency of PSâ ¡in darkness, actual photochemical efficiency, absorbed light energy per unit area, captured light energy per unit area, quantum yield of electron transport and PSâ activity, decreased the increase of K point and maintained chloroplast ultrastructure. The experiment in solar-greenhouse showed that FA obviously increased water use efficiency, promoted dry matter accumulation as well as the contents of Vc, soluble sugar, soluble protein and free amino acid, and decreased tannin content. Our results suggested that FA could improve the yield and quality of cucumber in solar greenhouse under drought stress.
Asunto(s)
Cucumis sativus , Sequías , Benzopiranos , Cucumis sativus/fisiología , Fotosíntesis , Hojas de la Planta/fisiología , AguaRESUMEN
To explore whether there is an interaction between melatonin (MT) and calcium (Ca2+) in regulating heat tolerance of plants, we analyzed the response of endogenous MT and Ca2+ to heat stress, and examined the effect of MT and Ca2+ on the reactive oxygen (ROS) accumulation, antioxidant system, and transcripts of heat shock factor (HSF) and heat shock proteins (HSPs) of cucumber seedlings under high temperature stress. Seedlings were foliar sprayed with 100 µmol·L-1 MT, 10 mmol·L-1 CaCl2, 3 mmol·L-1 ethylene glycol tetraacetic acid (EGTA, Ca2+ chelating agent) +100 µmol·L-1 MT, 0.05 mmol·L-1 chlorpromazine (calmodulin antagonist, CPZ) +100 µmol·L-1 MT, 100 µmol·L-1 p-chlorophenylalanine (p-CPA, inhibitor of MT) +10 mmol·L-1 CaCl2 or deionized water (H2O), respectively. The results showed that both endogenous MT and Ca2+ in cucumber seedlings were induced by high temperature stress. The seedlings treated with exogenous MT showed significant increases in the mRNA expression of calmodulin (CaM), calcium-dependent protein kinase (CDPK5), calcineurin B-like protein (CBL3) and CBL interacting protein kinase (CIPK2) compared with the control at normal temperature. The mRNA levels of tryptophane decarboxylase (TDC), 5-hydroxytryptamine-N-acetyltransferase (SNAT) and N-acetyl-5-hydroxytryptamine methyltransferase (ASMT), key genes of MT biosynthesis and endogenous MT content were also induced by Ca2+ in cucumber seedlings. Exogenous MT and CaCl2 alleviated the heat-induced oxidative damage through increasing antioxidant ability, reducing the accumulation of reactive oxygen species (ROS), and upregulating the mRNA abundances of HSF7, HSP70.1 and HSP70.11, as evidenced by mild thermal damage symptoms, lower heat injury index and electrolyte leakage under heat stress. The positive effect of MT-induced antioxidant capacity and mRNA expression of HSPs was removed by adding EGTA and CPZ in stressed seedlings. Similarly, the mitigating role of Ca2+ in the peroxidation damage to high temperature stress was reversed by p-CPA. These results suggested that both MT and Ca2+ could induce heat tolerance of cucumber seedlings, which had crosstalk in the process of heat stress signal transduction.
Asunto(s)
Cucumis sativus , Melatonina , Cucumis sativus/genética , Melatonina/farmacología , Calcio , Antioxidantes/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Calmodulina/genética , Calmodulina/metabolismo , Calmodulina/farmacología , Ácido Egtácico/farmacología , Cloruro de Calcio/metabolismo , Cloruro de Calcio/farmacología , Temperatura , Estrés Fisiológico , Plantones/fisiología , ARN Mensajero/metabolismo , ARN Mensajero/farmacologíaRESUMEN
Melatonin (MT) and nitric oxide (NO) are two multifunctional signaling molecules that are involved in the response of plants to abiotic stresses. However, how MT and NO synergize in response to cold stress affecting plants is still not clear. In this study, we found that endogenous MT accumulation under cold stress was positively correlated with cold tolerance in different varieties of cucumber seedlings. The data presented here also provide evidence that endogenous NO is involved in the response to cold stress. About 100 µM MT significantly increased the nitrate reductase (NR) activity, NR-relative messenger RNA (mRNA) expression, and endogenous NO accumulation in cucumber seedlings. However, 75 µM sodium nitroprusside (SNP, a NO donor) showed no significant effect on the relative mRNA expression of tryptophan decarboxylase (TDC), tryptamine-5-hydroxylase (T5H), serotonin-N-acetyltransferase (SNAT), or acetylserotonin O-methyltransferase (ASMT), the key genes for MT synthesis and endogenous MT levels. Compared with H2O treatment, both MT and SNP decreased electrolyte leakage (EL), malondialdehyde (MDA), and reactive oxygen species (ROS) accumulation by activating the antioxidant system and consequently mitigated cold damage in cucumber seedlings. MT and SNP also enhanced photosynthetic carbon assimilation, which was mainly attributed to an increase in the activity and mRNA expression of the key enzymes in the Calvin-Benson cycle. Simultaneously, MT- and SNP-induced photoprotection for both photosystem II (PSII) and photosystem I (PSI) in cucumber seedlings, by stimulating the PsbA (D1) protein repair pathway and ferredoxin-mediated NADP+ photoreduction, respectively. Moreover, exogenous MT and SNP markedly upregulated the expression of chilling response genes, such as inducer of CBF expression (ICE1), C-repeat-binding factor (CBF1), and cold-responsive (COR47). MT-induced cold tolerance was suppressed by 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO, a specific scavenger of NO). However, p-chlorophenylalanine (p-CPA, a MT synthesis inhibitor) did not affect NO-induced cold tolerance. Thus, novel results suggest that NO acts as a downstream signal in the MT-induced plant tolerance to cold stress.
RESUMEN
Chilling adversely affects the photosynthesis of thermophilic plants, which further leads to a decline in growth and yield. The role of melatonin (MT) in the stress response of plants has been investigated, while the mechanisms by which MT regulates the chilling tolerance of chilling-sensitive cucumber remain unclear. This study demonstrated that MT positively regulated the chilling tolerance of cucumber seedlings and that 1.0 µmolâ L-1 was the optimum concentration, of which the chilling injury index, electrolyte leakage (EL), and malondialdehyde (MDA) were the lowest, while growth was the highest among all treatments. MT triggered the activity and expression of antioxidant enzymes, which in turn decreased hydrogen peroxide (H2O2) and superoxide anion (O2 â -) accumulation caused by chilling stress. Meanwhile, MT attenuated the chilling-induced decrease, in the net photosynthetic rate (Pn) and promoted photoprotection for both photosystem II (PSII) and photosystem I (PSI), regarding the higher maximum quantum efficiency of PSII (Fv/Fm), actual photochemical efficiency (ΦPSII), the content of active P700 (ΔI/I0), and photosynthetic electron transport. The proteome analysis and western blot data revealed that MT upregulated the protein levels of PSI reaction center subunits (PsaD, PsaE, PsaF, PsaH, and PsaN), PSII-associated protein PsbA (D1), and ribulose-1,5-bisphosphate carboxylase or oxygenase large subunit (RBCL) and Rubisco activase (RCA). These results suggest that MT enhances the chilling tolerance of cucumber through the activation of antioxidant enzymes and the induction of key PSI-, PSII-related and carbon assimilation genes, which finally alleviates damage to the photosynthetic apparatus and decreases oxidative damage to cucumber seedlings under chilling stress.
RESUMEN
Salicylic acid (SA) has been proven to be a multifunctional signaling molecule that participates in the response of plants to abiotic stresses. In this study, we used cold-sensitive cucumber and cold-tolerant pumpkin as experimental materials to examine the roles of SA in root-shoot communication responses to aerial or/and root-zone chilling stress in own-root and hetero-root grafted cucumber and pumpkin plants. The results showed that pumpkin (Cm) rootstock enhanced the chilling tolerance of grafted cucumber, as evidenced by the observed lower levels of electrolyte leakage (EL), malondialdehyde (MDA), and higher photosynthetic rate (Pn) and gene expression of Rubisco activase (RCA). However, cucumber (Cs) rootstock decreased the chilling tolerance of grafted pumpkins. Cs/Cm plants showed an increase in the mRNA expression of C-repeat-binding factor (CBF1), an inducer of CBF expression (ICE1), and cold-responsive (COR47) genes and CBF1 protein levels in leaves under 5/25 and 5/5°C stresses, or in roots under 25/5 and 5/5°C stresses, respectively, compared with the Cs/Cs. Chilling stress increased the endogenous SA content and the activity of phenylalanine ammonia-lyase (PAL), and the increase in SA content and activity of PAL in Cs/Cm plants was much higher than in Cs/Cs plants. Transcription profiling analysis revealed the key genes of SA biosynthesis, PAL, ICS, and SABP2 were upregulated, while SAMT, the key gene of SA degradation, was downregulated in Cs/Cm leaves, compared with Cs/Cs leaves under chilling stress. The accumulation of SA in the Cs/Cm leaves was mainly attributed to an increase in SA biosynthesis in leaves and that in transport from roots under aerial and root-zone chilling stress, respectively. In addition, exogenous SA significantly upregulated the expression level of cold-responsive (COR) genes, enhanced actual photochemical efficiency (Φ PSII), maximum photochemical efficiency (F v/F m), and Pn, while decreased EL, MDA, and CI in grafted cucumber. These results suggest that SA is involved in rootstock-scion communication and grafting-induced chilling tolerance by upregulating the expression of COR genes in cucumber plants under chilling stress.