β-Glucosidase treatment and infestation by the rice brown planthopper Nilaparvata lugens elicit similar signaling pathways in rice plants

β-Glucosidase has been reported to induce the production of herbivore-induced plant volatiles. However, how it works remains unclear. Here, we investigated the levels of salicylic acid （SA）, jasmonic acid （JA）, ethylene, and H2O2, all of which are known signaling molecules that play important roles in induced plant defense in rice plants treated with β-glucosidase, and compared these to levels in plants infested by the rice brown planthopper Nilaparvata lugens （Stal）. Results showed that wounding and treatment by β-glucosidase increased the levels of SA, ethylene, and H2O2, but not JA, in all plants compared to control plants. The signaling pathways activated by β-glucosidase treatment are similar to those activated by an infestation by N. lugens, although the magnitude and timing of the signals elicited by the two treatments are different. This may explain why both treatments have similar volatile profiles and are equally attractive to the parasitoid Anagrus nilaparvatae Pang et Wang.     

Nitric oxide:A potential key point of the signaling network leading to plant secondary metabolite biosynthesis

The endogenous signaling network of plants plays important roles in mediating the exogenous factor-induced biosynthesis of secondary metabolites. Nitric oxide (NO) has emerged as a key signaling molecule in plants recently. Numerous studies demonstrated that the main signaling molecules such as salicylic acid (SA), jasmonic acid (JA), reactive oxygen species (ROS), and NO were not only involved in regulating plant secondary metabolite biosynthesis but also interacted to form a complex signaling network by mutual inhibition and/or synergy. The recent progress in the signal network of plant secondary metabolite biosynthesis has been discussed in this paper. Furthermore, we propose a hypothetical model to show that NO might act as a potential molecular switch in the signaling network leading to plant secondary metabolite biosynthesis.     

Promoter of soybean early nodulin gene<Emphasis Type="Italic">enod2B</Emphasis> is induced by rhizobial Nod factors in transgenic rice

Nod factors, which are signaling molecules produced by Rhizobia, are the principal determinants of host specificity in Rhizobium-legume symbiosis. Nod factors can elicit a number of characteristic developmental responses in the roots of legumes, such as depolarization of the membrane potential in epidermal cells, specific expression of early nodulin genes and changes in the flux of calcium in root hairs, deformation of root hairs, cell division in the root cortex and formation of the nodule primordinm. Whether the rice plant can respond to signaling molecules (i.e. Nod factors) is an important question, as it could establish the potential for symbiotic nitrogen fixation in rice. The promoter of the soybean (Glycine max) early nodulin gene Gmenod2B fused to the β-glucuronidase (GUS) reporter gene was used as a molecular marker to explore whether Nod factors can be recognized by rice cells as signaling molecules. Transgenic rice plants harboring the chimeric gene Gmenod2BP-GUS were obtained via an Agrobacterium tumefaciens-mediated system. NodNGR factors produced by a broad-host-range Rhizobium strain NGR234(pA28) were used as probes to investigate the activity of the Gmenod2B promoter in rice. Our results showed that the early nodulin gene Gmenod2B promoter was induced by NodNGR factors in transgenic rice, and that it was specifically expressed in rice plant roots. Moreover, GUS gene expression driven by the Gmenod2B promoter in transgenic rice was regulated by nitrogen status. These findings indicated that rice possessed the ability to respond to Nod factor signals, and that this signal transduction system resulted in activation of the Gmenod2B promoter. Thus, we predict that the Nod-factor inducible nodulin expression system, which is similar to Rhizobium-legume symbiosis, may also exist in rice.     

Involvement of Jasmonate-signaling pathway in the herbivore-induced rice plant defense

The expression patterns of eight defense- related genes in the herbivore-infested and jasmonate- treated (jasmonic acid, JA and its derivative MeJA) rice leaves were analyzed using RT-PCR. The results showed that Spodoptera litura Fabricius (Lepidoptera: Noctuidae) herbi-vory induced the expression of lipoxygenase (LOX) and al-lene oxide synthase (AOS) genes that are involved in the jasmonate-signaling pathway. Moreover, S. litura damage resulted in the expression of farnesyl pyrophosphate syn-thase (FPS), Bowman-birk proteinase inhibitor (BBPI), phenylalanine ammonia-lyase (PAL) and other rice defense- related genes that were also induced by aqueous JA treat-ment or gaseous MeJA treatment. These indicated that in rice leaves, the JA-related signaling pathway was involved in the S. litura-induced chemical defense. Mechanical damage and brown planthopper (BPH), Nilaparvata lugens (St錶) (Homoptera: Delphacidae) damage induced the expression of LOX gene, but both treatments did not induce the expression of AOS gene. However, BPH damage induced the expression of acidic pathogen-related protein 1 (PR-1a), Chitinase (PR-3), and PAL genes, which is involved in the salicylate- signaling pathway. It was suggested that salicylate-related signaling pathway or other pathways, rather than jas-monate-signaling pathway was involved in the BPH-induced rice plant defense.     

Obtaining transgenic rice resistant to rice fungal blast disease by controlled cell death strategy

The strategy of the two-component system,composed of Barnase and Barstar which encode RNase and a specific inhibitor to the RNase respectively, is adopted to obtain transgenic rice resistant to rice fungal blast disease. In this study, two chimeric promoters, induced by rice blast fungus pathogen (Magnaporthe grisea), are fused with Barnase respectively to construct two plant expression vectors, pWBNBS and pPBNBS together with the Barstar driven by CaMV 35S promoter. The resistance of the transgenic rice lines to rice blast fungus disease and rice blight disease are evaluated. The results show that (1) the expression of Barnase is induced in rice leaves when inoculated with the spores of Magnaporthe grisea; (2) the induced expression level of Barnase surpasses the level of Barstar, which elicits a similar hypersensitive response (HR) in the leaves, and the transgenic plant shows high resistance to the rice fungal blast disease; and (3) transgenic rice plants also show obvious resistance to rice bacterial blight disease. Taken together, these results suggest that the transgenic rice plants harboring this two-component system acquire relatively broad spectrum resistance against pathogens, especially high resistance to rice fungal pathogen.     

Nitric oxide involved in signal transduction of Jasmonic acid-induced stomatal closure of Vicia faba L.

Nitric oxide (NO) and Jasmonic acid (JA) are two key signaling molecules involved in many and diverse biological pathways in plants. Growing evidence suggested that NO signaling interacts with JA signaling. In this work, Our experiment showed that NO exists in guard cell of Vicia faba L., and NO is involved in signal transduction of JAinduced stomata closuring: ( i ) JA enhances NO synthesis in guard cell; ( ii ) both JA and NO induced stomatal closure, and had dose response to their effects; ( iU ) there are synergetic correlation between JA and lower NO concentration in regulation of stomatal movement; (iV) JA-induced stomatal closure was largely prevented by 2-phenyl-4,4,5,5-tetramethylimidazoline-l-oxyl-3-oxide (PTIO), a specific NO scavenger. An inhibitor of NO synthase (NOS) in mammalian cells, N^G-nitro-L-Arg-methyl eater (L-NAME) also inhibits plant NOS, repressing JA-induced NO generation and JA-induced stomatal closure. We presumed that NO mainly comes from NOS after JA treatment.     

Over-expression of an Arabi-dopsisd δ-OAT gene enhances salt and drought tolerance in transgenic rice

δ-OAT, ornithine-δ-aminotransferase, is the key enzyme involved in proline biosynthesis. In this study the Arabidopsis δ-OAT gene was transferred into rice (Oryza sativa L. ssp japonica cv. Zhongzuo 321), whose successful integration was demonstrated by PCR and Southern blot analysis. The over-expression of the gene in transgenic rice was also confirmed. Biochemical analysis showed that, under salt or drought stress conditions, proline contents in the leaves and roots in transgenic rice plants were 5- to 15-fold of those in non-transgenic controls. Under stress conditions, germinating rate of transgenic lines is higher than that of controls. Although the growth of rice plants tested were more and more retarded with the increasing of NaCI concentration, the transgenic plants grow faster compared to the controls under the same stress condition. Meanwhile, the resistance to KCl and MgSO4 stresses was also found enhanced in transgenie rice. Furthermore, the over-expression of δ-OAT also improved the yield of transgenic plants under stress conditions. The average yield per plant of transgenic lines increases about 12%--41% more than that of control line sunder 0.1 mol/L NaCI stress. These data indicated that the over-expression of δ-OAT, with the accumulation of proline, resulted in the enhancement of salt and drought tolerance and an increase of rice yield, which is of significance in agriculture.     

Systemic defense signaling in tomato

The wound-inducible expression of proteinase inhibitors （PIs） genes in tomato provides a powerful model system to elucidate the signal transduction pathway of systemic defense response. An increasing body of evidence indicates that systemin and jasmonic acid （JA） work in the same signaling pathway to activate the expression of Pls and other defense-related genes. However, little is known about how systemin and JA interact to regulate cell to cell communication over long distances. Genetic analysis of the systemin/JA signaling pathway in tomato plants provides a unique opportunity to dissect the mechanism by which peptide and oxylipin signals interact to coordinate systemic expression of defense-related genes. Previously, it has been proposed that systemin is the long-distance mobile signal for systemic expression of defense related genes. However, recent genetic approach provided new evidence that jasmonic acid, rather than systemin, functions as the systemic wound signal, and that the peptide systemin works to regulate the biosynthesis of JA.     

Cloning of two plastid division ftsZ genes from Nicotiana tabacum and their expression in E.coli

Two cDNAs of plastid division gene NtFtsZ1-1 and NtFtsZ1-2 are isolated from Nicotiana tabacum by RT-PCR and rapid amplification cDNA ends (RACE) method. Analysis of the deduced amino acid sequences encoded by NtFtsZ1-1 and NtFtsZ1-2 indicate that these two proteins possess the typical conservative motifs and GTP binding sites existing in all FtsZ proteins. The existence of putative plastid transit peptide in their N-terminal suggests that there are at least two transit-peptide containing FtsZ proteins in higher plants. Phylogenetic analysis based on amino acid sequences of FtsZ proteins also supports this interference. These two NtFtsZ genes demonstrate a similar expression pattern during the plant development, detected by Northern blot. Expression of NtFtsZ1-1 and NtFtsZ1-2 in E.coli interrupts the normal division process of host cells. These results suggest the diverse functions of FtsZ proteins in higher plants.     

Responses of rice growth to copper stress under free-air CO2 enrichment （FACE）

The responses of rice to the second degree contamination of copper were studied by pot experiments under free-air CO2 enrichment （FACE） with 570 μmol ·mol^-1 of CO2. The results showed that the content of copper in rice leaves was reduced with the CO2 concentration reaching 570 μmol· mol^-1 and this happened more significantly under the second degree contamination of copper. Under FACE, activities of superoxide dismutase （SOD） enzyme in rice leaves treated by copper contamination were induced, whereas the contents of glutathione （GSH） and glutathione disulfide （GSSG） had no significant difference from the control. In the presence of ambient CO2, activities of SOD enzyme treated by copper pollution were suppressed during the whole rice growth, however, the contents of GSH and GSSG were induced at tillering and jointing stages, and then restored to the control levels in later growth under the second degree contamination of copper. With the rice growing, the content of malondialdehyde （MDA） rises continuously, but there had been no significant difference between the treatments at the same growth stage. Further studies are needed on the response mechanism of rice to Cu stress under elevated CO2.     

Growth,physiological characteristics and ion distribution of NaCl stressed Alhagi sparsifolia seedlings

Alhagi sparsifolia is a leguminous perennial desert species that is plays an important role in dune stabilization and revegetation of degraded desert ecosystems. We investigated the effects of three different levels of salinity (50, 150, 250 mmol/L NaCl) on the growth, shoot photosynthetic parameters and salt distribution amongst different plant organs in one-year-old A. sparsifolia seedlings in a pot experiment over a 50 d period. The minimum (predawn) and maximum (midday) water potentials of A. sparsifolia seedlings decreased with the increase of external NaCl concentrations as a consequence of the osmotic or water deficit effect of saline solutions outside the roots. Salinity also reduced gas exchange parameters in A. sparsifolia, with seedlings subjected to salinity having lower photosynthesis rates and reduced stomatal conductances compared to the control. The reductions in photosynthetic rates in high salinity treatments of the A. sparsifolia seedlings were mainly caused by stomatal limitation. Consequently plants growing at greater external NaCl concentrations had significantly lower biomass accumulation compared to the control grown at 50 mmol/L. However, plants exposed to higher salinity were able to maintain growth throughout the experiment but allocated a greater proportion of biomass belowground. Plants exposed to higher external salinity levels had increased concentrations of Na+ and Cl- ions in shoots and roots, suggesting that A. sparsifolia seedlings were utilizing Na+ and Cl- as osmolytes to increase the cellular osmolality and decrease their water potential. We observed the greatest NaCl concentrations in the plants treated with 150 mmol/L NaCl indicating that there may be a threshold level of NaCl that can be tolerated by the plants. In conclusion our results indicate that A. sparsifolia seedlings are moderately salt tolerant. Photosynthetic gas exchange parameters were reduced by greater external salinity but the seedlings maintained substantial photosynthetic rates even under high salinity s     

Comparative Studies on Preparation of Large Plant DNA Fragments by Pulse Field Gel Electrophoresis

The preparation of large plant DNA fragments is extremely important to the construction of large insert DNA libraries (YAC, BAC, PAC and TAC). Although several techniques have been developed in each step of large plant DNA fragments preparation, the whole processing remains complicated and difficult. Based on authors‘ research experience and the recent worldwide development in this field, the following aspects are discussed in this paper: techniques of plant high molecular weight (HMW) DNA purification by pre-electrophoresis, the optimal conditions for the partial digestion of the HMW DNA by HindⅢ, the isolation effects of of large plant DNA fragments (100-400 kb) with different parameters of pulse field gel electrophoresis (PFGE), and the recovery of large DNA fragments. Through comparative studies, the advantages and disadvantages of each technique are discussed and some recommendations are proposed for preparing high quality large plant DNA fragments. The suggested techniques have been used in preparing the large DNA fragments of maize, rice, moss, laver, sea tangle and peach, and similar resuits are obtained among all the materials. This paper only reports the results using maize as material.     

Neurobiological disruptions induced in brains of the rats fed with mercury contaminated rice collected from experimental fields in Guizhou Province, China

Guizhou Province, located in the southwestern China, is an important mercury （Hg） production center. High levels of Hg were found in local environmental and biomass samples. The neurobiological disruptions induced in brains of rats fed with Hg polluted rice collected from two experimental plots in Guizhou Province were studied. The plots are close to the Hg mine of Wanshang area and a chemical plant （where Hg is used as a catalyst for the production of acetaldehyde） of Qingzhen area. Rats were fed for 30 d before toxicological end points were tested. The results showed that the accumulations of Hg in rat＇s brain of Qingzhen chemical plant group and Wanshang mine group were significantly higher than those of Shanghai group （control group） （P〈0.01）. The neural transmitters, including acetylcholine （Ach）, acetylcholine enzyme （AchE）, nitric oxide and nitricoxide synthase （NOS）, in rat＇s brain were significantly affected. Oxidative stress was observed in the exposure groups. The expression levels of the immediately early gene （lEG）, c-fos and c-FOS proteins were induced by the Hg polluted rice. It was noted that the lEG （c-fos） participated in the neurotoxic process induced by the Hg polluted rice. lEG response to Hg might be dependent on interactions between neural transmitter and oxidative damage, c.fos could be used as an effective index of detecting and assessing the neurotoxicity of Hg. The results suggested that it should be alert to mental health problem in human beings eating Hg-polluted food. More efforts are necessary to protect the local ecosystem and human health in the Hg polluted areas such as Wangshan and Qingzhen in Guizhou Province of China.