Combined transcriptome and proteome analysis revealed the molecular regulation mechanisms of zinc homeostasis and antioxidant machinery in tobacco in response to different zinc supplies.

Zinc (Zn) is an essential micronutrient for plants. Adequate regulation of Zn uptake, transport and distribution, and adaptation to Zn-deficiency stress or Zn-excess toxicity are crucial for plant growth and development. However, little has been done to understand the molecular responses of plants toward different Zn supply levels. In the present study, we investigated the growth and physiological responses of tobacco seedlings grown under Zn-completely deficient, Zn-limiting, Zn-normal, and Zn-4-fold sufficient conditions, respectively, and demonstrated that Zn deficiency/limitation caused oxidative stress and impaired growth of tobacco plants. Combined transcriptome and proteome analysis revealed up-regulation of genes/proteins associated with Zn uptake and distribution, including ZIPs, NAS3s, and HMA1s, and up-regulation of genes/proteins involved in regulation of oxidative stress, including SODs, APX1s, GPX6, and GSTs in tobacco seedlings in response to Zn deficiency/limitation, suggesting that tobacco possessed mechanisms to regulate Zn homeostasis primarily through up-regulation of the ZIPs-NAS3s module, and to alleviate Zn deficiency/limitation-induced oxidative stress through activation of the antioxidant machinery. Our results provide novel insights into the adaptive mechanisms of tobacco in response to different Zn supplies, and would lay a theoretical foundation for development of varieties of tobacco or its relatives with high tolerance to Zn-deficiency.

Expression and interaction of small heat shock proteins (sHsps) in rice in response to heat stress.

The inherent immobility of rice (Oryza sativa L.) limited their abilities to avoid heat stress and required them to contend with heat stress through innate defense abilities in which heat shock proteins played important roles. In this study, Hsp26.7, Hsp23.2, Hsp17.9A, Hsp17.4 and Hsp16.9A were up-regulated in Nipponbare during seedling and anthesis stages in response to heat stress. Subsequently, the expressing levels of these five sHsps in the heat-tolerant rice cultivar, Co39, were all significantly higher than that in the heat-susceptible rice cultivar, Azucena. This indicated that the expressive level of these five sHsps was positively related to the ability of rice plants to avoid heat stress. Thus, the expression level of these five sHsps can be regarded as bio-markers for screening rice cultivars with different abilities to avoid heat stress. Hsp18.1, Hsp17.9A, Hsp17.7 and Hsp16.9A, in the three rice cultivars under heat stress were found to be involved in one protein complex by Native-PAGE, and the interactions of Hsp18.1 and Hsp 17.7, Hsp18.1 and Hsp 17.9A, and Hsp17.7 and Hsp16.9A were further validated by yeast 2-hybridization. Pull down assay also confirmed the interaction between Hsp17.7 and Hsp16.9A in rice under heat stress. In conclusion, the up-regulation of the 5 sHsps is a key step for rice to tolerate heat stress, after that some sHsps assembled into a large hetero-oligomeric complex. In addition, through protein-protein interaction, Hsp101 regulated thiamine biosynthesis, and Hsp82 homology affected nitrogen metabolism, while Hsp81-1 were involved in the maintenance of sugar or starch synthesis in rice plants under heat stress. These results provide new insight into the regulatory mechanism of sHsps in rice.

Phosphoproteins regulated by heat stress in rice leaves.

BACKGROUND: High temperature is a critical abiotic stress that reduces crop yield and quality. Rice (Oryza sativa L.) plants remodel their proteomes in response to high temperature stress. Moreover, phosphorylation is the most common form of protein post-translational modification (PTM). However, the differential expression of phosphoproteins induced by heat in rice remains unexplored. METHODS: Phosphoprotein in the leaves of rice under heat stress were displayed using two-dimensional electrophoresis (2-DE) and Pro-Q Diamond dye. Differentially expressed phosphoproteins were identified by MALDI-TOF-TOF-MS/MS and confirmed by Western blotting. RESULTS: Ten heat-phosphoproteins were identified from twelve protein spots, including ribulose bisphos-phate carboxylase large chain, 2-Cys peroxiredoxin BAS1, putative mRNA binding protein, Os01g0791600 protein, OSJNBa0076N16.12 protein, putative H(+)-transporting ATP synthase, ATP synthase subunit beta and three putative uncharacterized proteins. The identification of ATP synthase subunit beta was further validated by Western-blotting. Four phosphorylation site predictors were also used to predict the phosphorylation sites and the specific kinases for these 10 phosphoproteins. CONCLUSION: Heat stress induced the dephosphorylation of RuBisCo and the phosphorylation of ATP-ß, which decreased the activities of RuBisCo and ATP synthase. The observed dephosphorylation of the mRNA binding protein and 2-Cys peroxiredoxin may be involved in the transduction of heat-stress signaling, but the functional importance of other phosphoproteins, such as H+-ATPase, remains unknown.

Differential regulation of proteins and phosphoproteins in rice under drought stress.

Drought is the largest constraint on rice production in Asia. Protein phosphorylation has been recognized as an important mechanism for environmental stress signaling. However, the differential expression of proteins and phosphoproteins induced by drought in rice is still largely unknown. In this paper, we report the identification of differentially expressed proteins and phosphoproteins induced by drought in rice using proteomic approaches. Three drought-responsive proteins were identified. Late embryogenesis abundant (LEA)-like protein and chloroplast Cu-Zn superoxide dismutase (SOD) were up-regulated by drought whereas Rieske Fe-S precursor protein was down-regulated. Ten drought-responsive phosphoproteins were identified: NAD-malate dehydrogenase, OSJNBa0084K20.14 protein, abscisic acid- and stress-inducible protein, ribosomal protein, drought-induced S-like ribonuclease, ethylene-inducible protein, guanine nucleotide-binding protein beta subunit-like protein, r40c1 protein, OSJNBb0039L24.13 protein and germin-like protein 1. Seven of these phosphoproteins have not previously been reported to be involved in rice drought stress. These results provide new insight into the regulatory mechanism of drought-induced proteins and implicate several previously unrecognized proteins in response to drought stress.

[Effects of different irrigation treatments on photosynthesis of Tieguanyin tea plants].

By using chlorophyll fluorescence kinetics technique, the effects of different irrigation intervals, i.e., 5 d (T1), 10 d (T2), 15 d (T3), 20 d (T4), and 25 d (T5), on the photosynthesis of 2-year Tieguanyin tea plants were investigated in the field, with no irrigation as the control. The results showed that the leaf water potential and chlorophyll content decreased with increasing irrigation interval, while the net photosynthesis (P) increased first and decreased then, reaching its highest value (15.55, micromol x m(-2) x s(-1)) in treatment T2. The ratio of the variable to maximal fluorescence (F(v)/F(m)), the variable fluorescence quenching (deltaF(v)), and the variable fluorescence quenching rate (deltaF(v)/F(o)) all got the highest in treatment T2, being 0.844, 342.5, and 4.03, respectively. The initial fluorescence (F(o)) decreased with increasing irrigation interval, while a reverse changing trend of F(o) was observed in the control, demonstrating that PS II reaction center was damaged by drought stress. In conclusion, irrigating per 10 d was favorable to the photosynthetic electron transport and CO2 assimilation of the tea plants, which would enhance their photosynthesis efficiency.

[Allelopathic potential and physiological mechanism of Oryza sativa L. under phosphorus deficiency stress].

In this paper, allelopathic rice PI312777 (PI) and non-allelopahtic rice Lemont (Le) were grown in the nutrient solution with two levels of phosphorus (P) supply, and their allelopathic effects on Echinochloa crus-galli L. were studied. The results showed that under phosphorous deficiency stress, the inhibitory effect of PI on E. crus-galli root growth had an obvious increase. After growing in the nutrient solution with low P supply (0.5 mg x L(-1)) for 5, 10 and 15 days, the inhibitory rate of PI on E. crus-galli root dry matter increased by 5.64%, 3.89% and 12.13%, respectively, while that of Le did not show the same trend, compared with normal P supply. With low P supply for 5, 10 and 15 days, PI allelopathic effect on the leaf POD activity of E. crus-galli increased by 20.19%, 15.47% and 6.68%, IAAoase activity increased by 18.08%, 17.71% and 12.50%, while NR activity increased by 13.89%, 18.60% and 2.10%, respectively, suggesting that under P deficiency, PI enhanced its allelopathic potential through two pathways, i.e., to increase the weed POD and IAAoase activities to slow down its growth rate, and to decrease the weed NR activity to affect its N uptake.

[Effects of different mycorrhizal fungi on physiological metabolism of tobacco seedlings].

Tobacco seedlings were inoculated with VA mycorrhizal fungi in natural soil. The results showed that compared with the control, the contents of nitrogen, phosphorus, potassium and chlorophyll, nitrate reductase activity, and protein in leaves were higher, malondialdehyde(MDA) and hydrogen peroxide(H2O2) decreased, while the activities of superoxide dismutase(SOD), catalase(CAT), and peroxidase(POD) increased. Meanwhile, seedlings were inoculated with two strains of ectomycorrhizal fungi respectively, and the above physiological indices trended the same changes. Moreover, the effect of strain Calvatia lilacina was higher than that of VA mycorrhizal fungi.

[Effects of NaCl stress on seedling growth and IAA metabolism of sweet potato and its relation to salt-tolerance].

Seedlings of three sweet-potato (Ipomoea batatas) cultivars with different salt-tolerance were stressed with different concentrations of NaCl during the seedling stage. Plant growth, leaf area growth, and IAA content decreased with increasing NaCl concentration, while the activities of POD and IAA oxidase increased. The extent of plant growth, leaf area growth and IAA content decrease and the extent of POD and IAA oxidase activity increase were larger in sensitive cultivar than those in moderately tolerant and tolerant cultivars. In addition, the content of chlorogenic acid increased under NaCl stress. Chlorogenic acid content reached its peak value at 170 mmol.L-1 of NaCl in sensitive and moderately tolerant cultivars and 225 mmol.L-1 of NaCl in tolerant cultivar. Chlorogenic acid content decreased with increasing NaCl concentration, but it was higher than that of the control.