Effects of Water Stress on the Protective Enzyme Activities and Lipid Peroxidation in Roots and Leaves of Summer Maize

A systematic study was conducted to determine the effects of water stress on the activities of protective enzymes and lipid peroxidation in maize. The results showed that, under water stress, the activities of superoxide dismutase （SOD）, catalase （CAT）, and peroxidase （POD） in leaves and roots increased sharply at prophase and metaphase growth stages, such as, male tetrad stage, but then declined towards the physiological maturity. The protective enzyme activities in roots were lower than those in leaves. The content of malondialdehyde （MDA） increased according to the severity of water stress. The content of MDA in roots was lower than that in leaves. The activities of protective enzymes and lipid peroxidation in roots were positively related to that in leaves with most of the correlation coefficients being significant. The content of soluble proteins in roots and leaves decreased with increasing drought stress. The ear characteristics deteriorated and the economic yields of maize decreased significantly under water stress. The main factors that caused reduction of yields were the decrease in the number of ear kernels and 100-kernel weight.     

Effects of Wounding and Exogenous Jasmonic Acid on the Peroxidation of Membrane Lipid in Pea Seedlings Leaves

The changes of malondialdehyde （MDA）, H2O2, and O2^7 content, or the activities of superoxide dismutase （SOD）, catalase （CAT）, ascrobate peroxidase （APX）, peroxidase （POD）, phenylalanine ammonia lyase （PAL）, and polyphenol oxidase （PPO） in pea seedlings （Pisum sativum L.） under wounding and treatment of exogenous jasmonic acid （JA） were investigated. The results showed that the activities of both phenylalanine ammonia lyase （PAL） and polyphenol oxidase （PPO） were significantly increased by wounding and application of JA. The metabolism of reaction oxidative species （ROS） was enhanced, especially O2^7 and H2O2 appeared to rapidly increase. The activities of antioxidant enzymes such as SOD, CAT, APX and POD were also increased. Treatment of JA of 1 or 10 μmol L^-1 could effectively induce plant defense response, and thus decrease the peroxidation of cell membrane lipid. However, high concentration of JA （100 μmol L^-1） resulted in unbalance of metabolism of ROS and promoted the peroxidation of cell membrane lipid. We thus suggested that JA, under the suitable concentration, could induce defense response of pea seedlings to wounding.     

Starch Accumulation and Enzyme Activities Associated with Starch Synthesis in Maize Kernels

The filling rate and the starch accumulation in developing maize kernel were analyzed. The changes of enzyme activities associated with sucrose metabolism and starch biosynthesis were investigated. The purpose is to discuss the enzymatic mechanisms responsible for starch synthesis. Two types of maize cultivars （Zea mays）, high starch maize （Feiyu 3） and normal maize （Yuyu 22）, were grown in a corn field. The factors involved in starch synthesis were performed during the growth period. The kernel filling rate, the sucrose content, the starch accumulating rates and the activities of SS （sucrose synthase）, GBSS （granule-bound starch synthase）, SBE （starch branching enzyme） of Feiyu 3, which has high starch content, were significantly higher than those of Yuyu 22, which has low starch content, after 10 DAP （days after pollination）. Correlation analysis indicated that ADPGPPase （ADP-glucose pyrophosphorylase） and DBE （starch debranching enzyme） were not correlated with the starch accumulating rates and the kernel filling rate, but the SS activity at the middle and late period were highly significantly correlated with the starch accumulating rates and the kernel filling rate. The GBSS activity was highly significantly correlated with the amylose accumulating rate, but not correlated with the kernel filling rate. The SBE activity was highly significantly correlated with the amylopectin accumulating rate and the kernel filling rate. It was not ADPGPPase and DBE, but SS was the rate-limiting factor of starch biosynthesis in developing maize kernels. GBSS had an important effect on amylose accumulation, and SBE had a significant effect on amylopectin accumulation.     

Effects of Terpinen-4-ol on Four Metabolic Enzymes and Polyphenol Oxidase （PPO） in Mythimna separta Walker

To study insecticidal mechanism of terpinen-4-ol, a main insecticidal composition in the essential oil of Sabina vulgaris, the 5th instar larvae of Mythimna separta, were investigated with terpinen-4-ol by topical application. The activities of phosphatase, glutathione S-transferase （GSTs）, cytochrome P450 （P450）, and polyphenol oxidase （PPO） of tested insects were determined in all poisoning stages, including exciting stage, convulsing stage, paralysis stage, and recover stage. The result showed that the activities of both acid phosphatase （ACP） and alkaline phosphatase （AKP） in treated insects were induced by terpinen-4-ol, but ACP was inhibited in paralysis stage. The activities of GSTs were inhibited in exciting stage, convulsing stage, and paralysis stage, but gradually recovered in recover stage. O-demethylase activity of cytochrome P450 was inhibited by terpinen-4-ol, and the inhibition rate in all poisoning stages were 26.27, 46.03, 80.24, and 90.22%, respectively. PPO activities were strongly inhibited by terpinen-4-ol both in vitro and in vivo. In conclusion, the activities of P450, GSTs, and PPO could have relation with toxicity of terpinen-4-ol against larvae of the Mythimna separta, but recover stage of the poisoning insects might be related to GSTs induced. As a new insecticide or synergist, terpinen- 4-ol has a potential value in field of insecticide resistance management.     

Effect of Dietary Alanyl-glutamine Supplementation on Growth Performance,Development of Intestinal Tract,Antioxidant Status and Plasma Non-specific Immunity of Young Mirror Carp（Cyprinus carpio L.）

Exogenous alanyl-glutamine（Aln-Gln） was evaluated for its effects on growth performance, intestinal structure and function, antioxidant status and non-specific immunity of young carp（Cyprinus carpio L.）. Six diets supplemented with 0, 2.5, 5.0, 7.5, 10.0, or 15.0 g · kg-1 of Aln-Gln were fed to fish for 12 weeks. Supplementation with 7.5, 10.0, or 15.0 g · kg-1 of Aln-Gln significantly increased weight gain rate（WGR）, protein efficiency ratio（PER）, but feed conservation rate（FCR） and survival were not affected（P〉0.05）. The intestinal fold height and number, digestive enzyme, Na＋, K＋-ATPase activities was found to be significantly high（P〈0.05） with increasing dietary Aln-Gln supplementation up to 7.5 g · kg-1, but there were no significant differences for Aln-Gln supplementation from 7.5 to 15.0 g · kg-1. The glutathione peroxidase（GPX） activity, glutathione（GSH）, superoxide dismutase（SOD） activity increased and malondialdehyde（MDA） levels decreased significantly（P〈0.05） in the intestine, hepatopancreas, plasma and muscles. The plasma complement-3（C3） and complement-4（C4） levels were significantly（P〈0.05） improved at 5.0 g · kg-1 level and decreased when over 7.5 g · kg-1. The plasma lysozyme（LSZ） activity increased significantly（P〈0.05） at 7.5, 10.0, or 15.0 g · kg-1 level. In summary, the results showed that Aln-Gln improved growth performance, development and function of the intestine, the activity of the antioxidant defense system and the plasma non-specific immunity of the carps. The optimal Aln-Gln level was 8.24 g · kg-1 diet for WGR based on broken-line regression model analysis.     

Study on Insecticidal Activities and Effect on Three Kinds of Enzymes by 5-Aminolevulinic Acid on Oxya chinensis

Insecticidal activities and effects on three enzymic activities caused by 5-aminolevulinic acid （ALA） on Oxya chinensis were studied. Fourth-instar nymphs of O. chinensis were treated with different doses ofALA （A1,250 mM; A2, 450 mM; A3,750 mM; A4, 1 000 mM）. Mortality and the activities of acetylcholinesterase （ACHE）, glutathione S-transferase （GSTs）, and glutathione peroxidase （GPx） were determinated. The mortality of O. chinensis rose with an increasing dose of ALA. The mortality of high-dose treatments A3 and A4 reached 66.19 and 80.21%, respectively. The value of LD50 was 3.61 （3.29-3.93） mg·g^-1 body weight （95% confidence interval）. Biochemical studies showed that the activities of AChE and GPx in the A4 treatment declined by 51.53 and 42.82% in the female, and 42.65 and 43.85% in the male compared to the control, respectively, and the degree of decline reached a significant level at P 〈 0.05. Meanwhile, the GSTs activities of O. chinensis enhanced with increasing dose of ALA. The GSTs activities of female and male O. chinensis in the A4 treatment remarkably increased by 171.05 and 97.42% compared to the control （P〈 0.05）. ALA had an obviously toxic effect on O. chinensis. Moreover, ALA caused the photoinactivation of AChE and GPx, which induced nerve transmission blocking and the capability to defend oxidation damage declining. Meanwhile, a high dose of ALA could activate GSTs, which caused a feedback inhibition of the insect to the phototoxic substance.     

Genotypic Difference in the Responses of Seedling Growth and Cd Toxicity in Rice （Oryza sativa L.）

The experiment was carried out to study the genotypic difference in the responses of seed germination, growth and physiological characters of rice seedlings to Cd toxicity. The result showed that the germination was slightly stimulated under low Cd concentration （0.01-1.5 mM Cd）, while severely depressed under higher Cd concentration （2.0 mM）. Rice seedlings exposed to 0.01 mM Cd showed slight increases in plant height, root volume, biomass and chlorophyll concentration. These parameters were significantly reduced when Cd level in the medium was increased to 0.5 mM, and meanwhile corresponding increase in superoxide dismutase （SOD） and peroxidase （POD） activities, and MDA （malondialdehyde） content was observed. However, SOD and POD activities declined when plants were exposed to 1 mM Cd when compared with those under 0.5 mM Cd. Cadmium addition lowered Fe, Cu and Mn concentrations in roots and shoots. There was significant genotypic difference in the response of these parameters to Cd stress. Under Cd stress, Xiushui 110 had the least inhibition of growth and increase in MDA content, higher shoot Cd concentration, and greatest increase in POD and SOD activities, indicating its higher tolerance to Cd toxicity, while Bing 9914 had the greatest reduction of growth, and Zn, Cu, Fe, and Mn contents, but greatest increase in MDA content, and least increase in activities of antioxidative enzymes, indicating its sensitivity to Cd toxicity.     

Responses of Antioxidant Enzymes to Chilling Stress in Tobacco Seedlings

Chilling is one of the major abiotic stresses limiting yield and quality of many important crops. For better understanding of chilling stress responses in tobacco （Nicotiana tabacum）, growth rate and antioxidant enzymes of seedlings in 2 tobacco cultivars, viz., MSk326 （chilling sensitive variety） and Honghuadajinyuan （HHDJY, chilling tolerant variety） at chilling temperature （5℃） were studied. The results showed that the relative growth rate in chilling period to that in recovery period was significantly higher in roots than that in shoots for both cultivars, suggesting that shoots growth was more easily affected by chilling stress. Chilling stress increased peroxidase （POD） activity and reduced superoxide dismutase （SOD） activity in shoots of HHDJY, and catalase （CAT） activity was little affected. In the roots of HHDJY, chilling stress increased SOD and CAT activities, and had little effect on POD activity. For MSk326, chilling treatment increased SOD activity in shoots and declined CAT activity in roots. MDA concentration in both varieties was increased under the chilling stress, while it was decreased after seedlings were recovered growth for 4 d at normal temperature （25℃）. It showed that tobacco seedlings might have the capacity of recovering from chilling injury for a short term, The relationship between the growth rate and antioxidant enzyme activity was analyzed by stepwise regression. It was found that there was a close relationship between relative growth rate of tobacco seedlings and CAT activity under chilling stress condition and regression equations containing CAT could be used in predicting seedling growth rate of tobacco under chilling stress condition.     

Accumulation of Carbohydrate and Regulation of 14-3-3 Protein on Sucrose Phosphate Synthase （SPS）Activity in Two Tomato Species

To explore the differences of carbohydrate metabolism in two tomato species and discuss the possible regulation of 14-3-3 proteins on the sucrose phosphate synthase （SPS） activity, we determined the contents of soluble sugar and starch through high performance liquid chromatography （HPLC）. The activities of sugar-metabolizing enzymes were assayed in desalted extract, and the relative expression levels of related genes in sugar metabolism were determined though real-time RT-PCR. The results indicated that glucose and fructose were mainly accumulated during the maturation of the fruit because of the high acid invertase （AI） and neutral invertase （NI） in Micro-Tom （Solanum lycopersicum） fruit, while in Solanum chmielewskii fruit, SPS which went along with the change of sucrose content led to the rapid sucrose increase during the fruit ripening. TFT1 and TFT10, belonging to 14-3-3 protein in tomato, were likely to down-regulated SPS activity during young and intumescence period.     

The Sublethal Effects of Chlorantraniliprole on Helicoverpa armigera (Lepidoptera: Noctuidae)

Sublethal effects of chlorantraniliprole to Helicoverpa armigera (Hübner) larvae were evaluated through exposure of third instar larvae to the insecticide incorporated into an artificial diet. When larvae were fed on the insecticide-containing diet for 7d, the LC10 , LC20 , LC40 , and LC50 values were 3.790, 7.977, 21.577, and 33.121 μg active ingredient L-1, respectively. Chlorantraniliprole at sublethal concentrations significantly reduced the larval body mass, emergence ratio, adult longevity and egg hatching rate in both the parental and offspring generations. The pupation and copulation rate in the parental generation and the pupal mass in the offspring also strongly decreased. Reproduction was seriously disturbed in both the parental and offspring groups even when only one of the partners was exposed to chlorantraniliprole at larval stages. However, at the lowest concentration of exposure (LC10 ), offspring fecundity was strongly reduced only when both partners were exposed. The net reproductive rate (R0 ), intrinsic rate of increase (rm ), and gross reproduction rate (GRR) at LC20 , LC40 concentrations were significantly lower than that of the control. Post-exposure effects also included an extended larval developmental time and increased male proportion in both generations. The doubling time (Dt) at LC20 , LC40 concentrations as well as gross reproduction rate (GRR) at LC10 concentration were also significantly increased. Chlorantraniliprole might have significant effects on H. armigera population dynamics even at sublethal concentrations on both parental and offspring generations.     

Paraquat Resistance in Leaf Discs of PAAG12-IPTModified Gerbera Is Related to the Activities of Superoxide Dismutase, Catalase, and Dehydroascorbate Reductase

In this paper, using in vitro leaf disc culture system, the changes of contents of chlorophylls, carotenoids, soluble protein, thiobarbituric acid reactive substance （TBARS）, and activities of antioxidant enzymes were investigated during the incubation of leaf discs of PSAG12-IPT modified gerbera in 0, 25, 50 μmol L^-1 paraquat （PQ） under continuous light intensity of 130 0tool m-2 s-1, compared with the control plant （wild type）. The results showed that PQ treatment significantly decreased the contents of chlorophylls, carotenoids, and soluble protein, therefore, promoted leaf senescence. However, the decreases in the leaf discs of modified gerbera were considerably smaller. The activities of superoxide dismutase （SOD）, catalase （CAT）, and dehydroascorbate reductase （DHAR） were significantly increased by PQ treatment and with the increasing of PQ concentration, particularly in the modified plants. The activities of ascorbate peroxidase （APX） and guaiacol peroxidase （GPX） could not be detected in the leaf discs of PQ treatments, which suggested that they were labile to the oxidative stress induced by PQ. As a product of lipid peroxidation, TBARS significantly increased in content with the increase of PQ concentration, while its concentration in the modified plants was significantly lower than that of control plants. Therefore, it could be concluded that the chimeric gene PSAG12-IPTtransfonnexi gerbera leaves had higher antioxidative potential, thus causing the delay of senescence under oxidative stress induced by PQ.     

Antioxidant Defense System in Wheat （Triticum aestivum L.） Seedlings under Heat Stress and Revival Conditions

The present investigation was carried out to investigate the effect of heat stress and revival on some antioxidative enzymes and metabolites in leaves of the wheat （Triticum aestivum L.） seedlings of heat susceptible （cv. WH 147 and HS 277） and heat tolerant （cv. WH 1021 and HW 2045） cultivars. Seven days old seedlings grown at 25 ℃ were exposed to 40 ℃ for 6 h and these seedlings were again brought to 25 ℃. The observations were recorded in the leaves of control, stressed and revived seedlings on 2nd and 4th day of revival. For the selection ofthermo-tolerant cultivars, screening of the thirty-six cultivars was done based on wilting of primary leaf and values of chlorophyll fluorescence. The MDA （malondialdehyde） and H2O2 concentration in leaves of wheat seedlings increased at the high temperature. There was enhancement in the activities of antioxidative enzymes, viz. CAT （catalase）, POX （peroxidase）, GR （glutathione reductase） and APX （ascorbate peroxidase） in leaves of the tolerant and susceptible cultivars under heat stress, however, higher percent increase was observed in tolerant cultivars. Heat stress increased the SOD （superoxide dismutase） activity in tolerant cultivars but activity declined in susceptible cultivars. On revival, the activities of the CAT, POX and GR declined in comparison to stressed seedlings but remained higher as compared to control. Ascorbate peroxidase activity remained higher on 2nd day and 4th day of revival in all the cultivars.     

The Effects of Arbuscular Mycorrhizal Fungi on Reactive Oxyradical Scavenging System of Tomato Under Salt Tolerance

The effects of arbuscular mycorrhizal fungi （AMF）, Glomus mosseae, on oxygen radical scavenging system of tomato under salt stress were studied in potted culture experiments. The response of tomato （Lycopersieon eseulentum L.） cultivar Zhongza 9 seedlings with AMF inoculation and control to salt stress （0, 0.5 and 1.0% NaCl solution, respectively） was investigated. The results showed that the salt stress significantly reduced the dry matter content of roots, stems and leaves, and also the leaf area as compared with the control treatment. However, arbuscular mycorrhizal-inoculated （AM） significantly improved the dry matter and the leaf area in the salt-stressed plants. The effect of AMF on dry matter was more pronounced in aerial bromass than in root biomass which might be due to AM colonization. The activities of SOD, POD, ASA-POD, and CAT in leaves and roots of mycorrhizal and non-mycorrhizal treatment of tomato plants were increased and had different rules under different NaCl concentrations （solution of 0, 0.5 and 1% NaCl）, but all enzymes had a rise in the beginning of treatment under salt stress conditions. The AMF did not change the rule of tomato itself under salt stress, but AMF increased these enzyme activities in different levels. The AMF treatment significantly increased SOD, POD and ASA-POD activities in leaves and roots, whereas it had little effects on CAT in root. O2- production rate and MDA content in leaves increased continuously, which showed a positive line correlation with salt stress concentration. O2- production rate and MDA content in tomato plants significantly decreased by AM treatment compared with nonmycorrhizal treatment. In conclusion, AM could alleviate the growth limitations imposed by saline conditions, and thereby play a very important role in promoting plant growth under salt stress in tomato.     

Coronatine Induces an Accumulation of Anthocyanin and Starch in Purple-fleshed Sweetpotato （ Ipomoea batatas Lam.）

[Objective] The objective of this research was to examine the effects of COR on anthocyanin and starch content in storage roots of two PFS genotypes, and to explore the relationships between anthocyanin synthesis and starch accumula- tion. [Method] A field experiment was carried out to determine the changes in yielc components, yield, contents of anthocyanin and starch, activities of phenylalanine ammonia-lyase （PAL） and adenosine 5-diphosphate glucose pyrophosphorylase （AG- Pase） in two genotypes of PFS （Ipomoea batatas L., var. ＇Ayamurasaki＇ and ＇Jishu18＇）. [Result] The application of COR significantly increased starch and antho- cyanin content in storage roots of Jishu18 across developmental stages by inducing the activities of PAL and AGPase, and finally enhanced yield by promoting fresh weight of storage roots. Ayamurasaki was insensitive to treatment with COR al- though its PAL activity temporally increased. The starch and anthocyanin content of Aya, and the anthocyanin content of Jishu18 increased progressively across devel- opmental stages with or without COR application, but the starch content of Jishu18 increased initially, then decreased before increasing again without application of COR. Treatment with COR reduced downward trend of starch accumulation in Jishu18. Thus, the effect of COR on accumulation of anthocyanin and starch in storage roots of PFS differs according to genotypes. [Conclusion] The application of 0.05 μmol/L COR may increase starch and anthocyanin content in PFS genotypes with lower starch and anthocyanin content in storage roots.     

Leaf Responses of Micropropagated Apple Plants to Water Stress： Changes in Endogenous Hormones and Their Influence on Carbohydrate Metabolism

The changes in the concentrations of endogenous hormones and their influence on carbohydrate metabolism in leaves of micropropagated Fuji apple plants were studied under water deficiency stress. The results showed that water stress induced a rapid increase in the concentration of abscisic acid （ABA） and led to a decrease in concentrations of both zeatin and gibberellins （GAs）. The concentration of indole-3-acetic acid （IAA） changed in an independent manner, which was not correlated with the different levels of water stress. With regard to the carbohydrates, the contents of sorbitol and sucrose increased, whereas the content of starch decreased. The increase in the concentration of ABA was significantly correlated with both the increase in the activity of aldose-6-phosphate reductase （A6PR） and the decrease in the activity of sorbitol dehydrogenase （SDH）, indicating that ABA played a regulatory role in sorbitol metabolism. The concentration of ABA was positively correlated to the activity of sucrose-phosphate synthase （SPS） but negatively correlated to the activities of acid invertase （AI） and ADP-glucose-pyrophosphorylase （ADPGppase） in water-stressed plants, which indicated that ABA promoted sucrose synthesis and inhibited sucrose degradation and starch synthesis at the same time. Under conditions of water stress, the decrease in the level of zeatin was accompanied by a decrease in the activities of SDH and ADPGPPase. GAs concentration showed positive correlation with ADPGPPase activity. IAA showed no significant correlation with any of the enzymes tested in this study. The results of this study suggested that ABA might be one of the key factors regulating the distribution of carbohydrates under water stress. The metabolism of sorbitol and starch under conditions of water stress might be regulated by the combined action of many plant hormones.     

Effect of Exogenous MJA Treatment of Tea Plants on the Growth of Geometrid Larvae

The effect of tea plant Camellia sinensis induced by exogenous methyl jasmonate （MJA） on lipoxygenase （LOX）, polyphenol oxidase （PPO） and proteinase inhibitor （PI） activity in the leaves of tea plants, as well as the growth and midgut proteinase activity of the geometrid Ectropis obliqua larvae were studied. MJA significantly induced LOX, PPO and PI activity in leaves of tea plants. When geometrid larvae have fed on leaves of tea plants treated with MJA, the activities of the high alkaline trypsin-like enzyme and chymotrypsin-like enzyme in their midgut were significantly inhibited, but the activities of the low alkaline trypsin-like enzyme in their midgut were unaffected, leading to imbalance between different types of proteinase activity in the midgut of the larvae and in turn, the growth were inhibited. These chains of response may be an important mechanism of the direct resistance induced by MJA-treatment of tea plant on geometrid larvae.     

Low Root Zone Temperature Exacerbates the Ion Imbalance and Photosynthesis Inhibition and Induces Antioxidant Responses in Tomato Plants Under Salinity

The combined effects of salinity with low root zone temperature （RZT） on plant growth and photosynthesis were studied in tomato （Solanum lycopersicum） plants. The plants were exposed to two different root zone temperatures （28/20℃, 12/8℃, day/night temperature） in combination with two NaC1 levels （0 and 100 mmol L-l）. After 2 wk of treatment, K＋ and Na~ concentration, leaf photosynthetic gas exchange, chlorophyll fluorescence and leaf antioxidant enzyme activities were measured. Salinity significantly decreased plant biomass, net photosynthesis rate, actual quantum yield of photosynthesis and concentration of K＋, but remarkably increased the concentration of Na＋. These effects were more pronounced when the salinity treatments were combined with the treatment of low RZT conditions. Either salinity or low RZT individually did not affect maximal efficiency of PSII photochemistry （Fv/Fm）, while a combination of these two stresses decreased Fv/Fm considerably, indicating that the photo-damage occurred under such conditions. Non-photochemical quenching was increased by salt stress in accompany with the enhancement of the de-epoxidation state of the xanthophyll cycle, in contrast, this was not the case with low RZT applied individually. Salinity stress individually increased the activities of SOD, APX, GPOD and GR, and decreased the activities of DHAR. Due to the interactive effects of salinity with low RZT, these five enzyme activities increased sharply in the combined stressed plants. These results indicate that low RZT exacerbates the ion imbalance, PSII damage and photosynthesis inhibition in tomato plants under salinity. In response to the oxidative stress under salinity in combination with low RZT, the activities of antioxidant enzymes SOD, APX, GPOD, DHAR and GR were clearly enhanced in tomato plants.