jun Zhu

Effects of osmotic stress on antioxidant enzymes activities in leaf discs of PSAG12-IPT modified gerbera

Leaf senescence is often caused by water deficit and the chimeric gene PSAG12-IPT is an auto-regulated gene delaying leaf senescence. Using in vitro leaf discs culture system, the changes of contents of chlorophylls, carotenoids, soluble protein and thiobarbituric acid reactive substance (TBARS) and antioxidant enzymes activities were investigated during leaf senescence of PSAGl2-IPT modified gerbera induced by osmotic stress compared with the control plant (wild type). Leaf discs were incubated in 20%, 40% (w/v) polyethylene glycol (PEG) 6 000 nutrient solution for 20 h under continuous light [130 μmol/(m2·s)]. The resultsshowed that the contents of chlorophylls, carotenoids and soluble protein were decreased by osmotic stress with the decrease beingmore pronounced at 40% PEG, but that, at the same PEG concentration the decrease in the transgenic plants was significantly lower than that in the control plant. The activities of superoxide dismutase (SOD), catalases (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and dehydroascorbate reductase (DHAR) were stimulated by PEG treatment. However, the increaseswere higher in PSAG12-IPT transgenic plants than in the control plants, particularly at 40% PEG treatment. Lipid peroxidation (TBARS content) was increased by PEG treatment with the increase being much lower in transgenic plant than in the control plant. It could be concluded that the increases in the activities of antioxidant enzymes including SOD, CAT, APX, GPX and DHAR were esponsible for the delay of leaf senescence induced by osmotic stress.

Effect of Nitrogen and Sulfur Supply on Glucosinolates in Brassica Campestris ssp. chinensis

Glucosinolates (GSs) are a group of plant secondary metabolites containing abundant nitrogen (N) and sulfur (S) mainly in Brassica and have the beneficial effects on human health including anti-carcinogenic, cholesterol-reducing and other pharmacological effects. The objective of this study was to investigate the effect of different concentrations of N (5, 10, and 20 mmol L^(-1), denoted by N5, Nl0 and N20) and S (0.5, 1, and 2 mmol L^(-1), denoted by S0.5, S1 and S2) on the yield and GSs in pakchoi (Brassica campestris L. ssp. chinensis var. communis) in hydroponics. Results showed that NI0 and N20 significantly enhanced the yield compared with N5, however, N20 had a negative effect relative to N10. Only with Nl0 and N20 low S supply (S0.5) reduced the yield. The concentrations of aliphatic GSs, aromatic GS and total GSs were enhanced by N5 and indoly1 GSs were enhanced by N20. S2 enhanced the concentration of individual GS and total GSs. The concentrations of indoly1 GSs were maximized in N20S2 treatment, whereas the highest concentrations of aliphatic GSs, aromatic GS and total GSs were found in N5S2 treatment. Effects of N and S on aliphatic GSs were higher than on indolyl GSs. The results suggest that the accumulation of aliphatic GSs and aromatic GS could be enhanced by low N and high S and restricted by high N while that of indolyl GSs could be enhanced by high N and high S.

Combined Effects of Excess Mn and Low pH on Oxidative Stress and Antioxidant Enzymes in Cucumber Roots

The effects of excess Mn on oxidative stress and antioxidant enzymes in cucumber roots under different pH (pH 4.5 and 6.5) were studied. The results indicated that lipid peroxidation was increasingly serious with the increased concentration of Mn, especially under low pH. As a result, the growth of cucumber roots and shoots was significantly inhibited. Excess Mn led to an elevation in activities of Mn-SOD and Fe-SOD, which played an important role in removing O2(superscript -). CAT was sensitive to excess Mn, and its activity significantly decreased under excess Mn and low pH, while activities of GPX, APX, DHAR and GR increased under low pH and to some extent under excess Mn, which indicated their important roles in scavenging reactive oxygen in tolerance to low pH and excess Mn of cucumber roots.

Isolation and expression of glucosinolate synthesis genes CYP83A1 and CYP83B1 in Pak Choi (Brassica rapa L. ssp. chinensis var. communis (N. Tsen & S.H. Lee) Hanelt).

CYP83A1 and CYP83B1 are two key synthesis genes in the glucosinolate biosynthesis pathway. CYP83A1 mainly metabolizes the aliphatic oximes to form aliphatic glucosinolate and CYP83B1 mostly catalyzes aromatic oximes to synthesis corresponding substrates for aromatic and indolic glucosinolates. In this study, two CYP83A1 genes named BcCYP83A1-1 (JQ289997), BcCYP83A1-2 (JQ289996) respectively and one CYP83B1 (BcCYP83B1, HM347235) gene were cloned from the leaves of pak choi (Brassica rapa L. ssp. chinensis var. communis (N. Tsen & S.H. Lee) Hanelt) “Hangzhou You Dong Er” cultivar. Their ORFs were 1506, 1509 and 1500 bp in length, encoding 501, 502 and 499 amino acids, respectively. The predicted amino acid sequences of CYP83A1-1, CYP83A1-2 and CYP83B1 shared high sequence identity of 87.65, 86.48 and 95.59% to the corresponding ones in Arabidopsis, and 98.80, 98.61 and 98.80% to the corresponding ones in Brassica pekinensis (Chinese cabbage), respectively. Quantitative real-time PCR analysis indicated that both CYP83A1 and CYP83B1 expressed in roots, leaves and petioles of pak choi, while the transcript abundances of CYP83A1 were higher in leaves than in petioles and roots, whereas CYP83B1 showed higher abundances in roots. The expression levels of glucosinolate biosynthetic genes were consistent with the glucosinolate profile accumulation in shoots of seven cultivars and three organs. The isolation and characterization of the glucosinolate synthesis genes in pak choi would promote the way for further development of agronomic traits via genetic engineering.

Variation in glucosinolates in pak choi cultivars and various organs at different stages of vegetative growth during the harvest period.

Glucosinolates (GSs) play an important role in plant defense systems and human nutrition. We investigated the content and composition of GSs in the shoots and roots of seven cultivars of pak choi. We found that ‘Si Yue Man’ had the highest total and aliphatic GS contents in the shoots and the highest benzenic GS content in the roots, ‘Shanghai Qing’ contained the highest amounts of benzenic and total GS contents in the roots, while ‘Nanjing Zhong Gan Bai’ had the lowest benzenic, indole, and total GS contents in both the shoots and roots. Therefore, the ‘Si Yue Man’ cultivar appears to be a good candidate for future breeding. Variation between the shoots and roots was also examined, and a significant correlation among the total, aliphatic, and some individual GSs was found, which is of value in agricultural breeding. GS concentrations of the leaf, petiole, and root increased dramatically during the period of rapid growth of the dry matter of the plant 10 to 20 d after transplantation, reaching peak values on Day 20 and decreasing on Day 25. We conclude that the pak choi should be harvested and consumed from 20 to 25 d after transplantation to take advantages of the high GS content in the plant.

Cloning and functional analysis of a novel ascorbate peroxidase (APX) gene from Anthurium andraeanum

An 888-bp full-length ascorbate peroxidase (APX) complementary DNA (cDNA) gene was cloned from Anthurium andraeanum, and designated as AnAPX. It contains a 110-bp 5′-noncoding region, a 28-bp 3′-noncoding region, and a 750-bp open reading frame (ORF). This protein is hydrophilic with an aliphatic index of 81.64 and its structure consisting of α-helixes, β-turns, and random coils. The AnAPX protein showed 93%, 87%, 87%, 87%, and 86% similarities to the APX homologs from Zantedeschia aethiopica, Vitis pseudoreticulata, Gossypium hirsutum, Elaeis guineensis, and Zea mays, respectively. AnAPX gene transcript was measured non-significantly in roots, stems, leaves, spathes, and spadices by real-time polymerase chain reaction (RT-PCR) analysis. Interestingly, this gene expression was remarkably up-regulated in response to a cold stress under 6 °C, implying that AnAPX might play an important role in A. andraeanum tolerance to cold stress. To confirm this function we overexpressed AnAPX in tobacco plants by transformation with an AnAPX expression construct driven by CaMV 35S promoter. The transformed tobacco seedlings under 4 °C showed less electrolyte leakage (EL) and malondialdehyde (MDA) content than the control. The content of MDA was correlated with chilling tolerance in these transgenic plants. These results show that AnAPX can prevent the chilling challenged plant from cell membrane damage and ultimately enhance the plant cold tolerance.

Paraquat Resistance in Leaf Discs of PSAG12-IPT Modified 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 P(subscript SAG12)-IPT modified gerbera in 0, 25, 50 μmol L^(-1) paraquat (PQ) under continuous light intensity of 130 μmol 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 P(subscript SAG12)-IPT transformed gerbera leaves had higher antioxidative potential, thus causing the delay of senescence under oxidative stress induced by PQ.

Genome-Wide Identification and Analysis of Polygalacturonase Genes in Solanum lycopersicum

Polygalacturonase (PG), a large hydrolase family in plants, is involved in pectin disassembly of the cell wall in plants. The present study aims to characterize PG genes and investigate their expression patterns in Solanum lycopersicum. We identified 54 PG genes in the tomato genome and compared their amino acid sequences with their Arabidopsis counterpart. Subsequently, we renamed these PG genes according to their Arabidopsis homologs. Phylogenetic and evolutionary analysis revealed that these tomato PG genes could be classified into seven clades, and within each clade the exon/intron structures were conserved. Expression profiles analysis through quantitive real-time polymerase chain reaction (qRT-PCR) revealed that most SlPGs had specific or high expression patterns in at least one organ, and particularly five PG genes (SlPG14, SlPG15, SlPG49, SlPG70, and SlPG71) associated with fruit development. Promoter analysis showed that more than three cis-elements associated with plant hormone response, environmental stress response or specific organ/tissue development exhibited in each SlPG promoter regions. In conclusion, our results may provide new insights for the further study of PG gene function during plant development.