Comparative expression analysis of three genes from the Arabidopsis vacuolar Na^＋/H^＋ antiporter （AtNHX） family in relation to abiotic stresses

Na~ /H~ antiporters (NHX) are ubiquitous transmembrane proteins that play a key role in salt tolerance of plants. In this study, the sequence of 3 Arabidopsis NHX gene (AtNHX2―4) were compared with other AtNHX members. Putative cis-elements analysis identified elements that have been associated with stress responses. The activities of the promoters AtNHX2―4 were studied in transgenic plants carrying corresponding promoter-β-glucuronidase (GUS) fusions. The AtNHX2 promoter-GUS analysis indicated that AtNHX2 was expressed in constitutive pattern with high GUS activity in roots and leaves. AtNHX2 promoter activity was not up-regulated by NaCl or abscisic acid (ABA), in contrast to the AtNHX1 promoter which was previously studied. The AtNHX3 and AtNHX4 promoters showed tissue-specific activities. Strong GUS activity was detected in roots and vascular bundles of the stele in plants carry-ing an AtNHX4 promoter-GUS fusion, and GUS activity increased under salt stress suggesting a func-tion related to salt tolerance. Transgenic plants carrying the AtNHX3 promoter-GUS fusion showed strong GUS activity in petals, stamens and tops of siliques, suggesting a possible role of AtNHX3 in flower and seed development. Results of histochemical analysis suggested that AtNHX2―4 are involved in divergent functions and are differentially regulated under abiotic stress. The structure of AtNHX4 was predicted to include 12 transmembrane regions and a NHX domain. Overexpression of AtNHX4 in Arabidopsis transgenic lines confers greater salt tolerance than in wild type plants. These results suggest that AtNHX4 may encode a putative vacuolar NHX that plays an important role in salt tolerance.     

Functional characterization and mapping of two MADS box genes from peach （Prunus persica）

Previously an AGAMOUS gene homologue PpMADS4 and a FRUITFULL gene homologue PpMADS6 were isolated from peach （Prunus persica）, and both genes were shown to express in the developing floral and fruits. To gain insight into their function, the two genes were constitutively expressed in Arabidopsis thaliana and their effects on plant growth and floral organ development were studied in this work. The transgenic plants all displayed early flowering and conversion of inflorescence to floral meristem. However, the two genes had different effects on the floral organ structures in A. thaliana. The transgenic plants overexpressing PpMADS4 displayed homeotic conversion of floral organs, and particularly the perianth abscission was inhibited. The plants overexpressing PpMADS6 showed early flowering, produced higher number of carpels, petals, and stamens than nontransgenic plants, and pod shatter was prevented; significantly, the transgenic plants yielded more than one siliques from a single flower. A SSR molecular marker was developed for PpMADS4, and it was then assigned into the G5 linkage group of Prunus sp. Both PpMADS4 and PpMADS6 genes were located at the same region in the G5 linkage group. Our results showed the potential application of these two MADS box genes for crop and fruit tree improvement.     

Functional characterization and mapping of two MADS box genes from peach (Prunus persica)

Previously an AGAMOUS gene homologue PpMADS4 and a FRUITFULL gene homologue PpMADS6 were isolated from peach (Prunus persica), and both genes were shown to express in the developing floral and fruits. To gain insight into their function, the two genes were constitutively expressed in Arabidopsis thaliana and their effects on plant growth and floral organ development were studied in this work. The transgenic plants all displayed early flowering and conversion of inflorescence to floral meristem. However, the two genes had different effects on the floral organ structures in A. thaliana. The transgenic plants overexpressing PpMADS4 displayed homeotic conversion of floral organs, and par- ticularly the perianth abscission was inhibited. The plants overexpressing PpMADS6 showed early flowering, produced higher number of carpels, petals, and stamens than nontransgenic plants, and pod shatter was prevented; significantly, the transgenic plants yielded more than one siliques from a single flower. A SSR molecular marker was developed for PpMADS4, and it was then assigned into the G5 linkage group of Prunus sp. Both PpMADS4 and PpMADS6 genes were located at the same region in the G5 linkage group. Our results showed the potential application of these two MADS box genes for crop and fruit tree improvement.     

Impact of high dose Kr+ ion irradiation on the corrosion behavior and microstructure of Zircaloy-4

In order to investigate the ion irradiation effect on the corrosion behavior and microstructure of Zircaloy-4, the Zircaloy-4 film were prepared by electron beam deposition on the Zircaloy-4 specimen surface and irradiated by Kr ions using an accelerator at an energy of 300 keV with the dose from 1×1015 to 3×1016 ions/cm2. The post-irradiation corrosion tests were conducted to rank the corrosion resistance of the resulting specimens by potentiodynamic polarization curve measurements in a 0.5 mol/L H₂SO₄ water solution at room temperature. Transmission electron microscopy (TEM) was employed to examine the microstructural change in the surface. The potentiodynamic tests show that with the irradiation dose increasing, the passive current density, closely related to the surface corrosion resistance, decreases firstly and increases subsequently. The mechanism of the corrosion behavior transformation is due to the amorphous phase formation firstly and the amorphous phase destruction and the polycrystalline structure formation in the irradiated surface subsequently.     

ENVIRONMENTAL ISSUES IN THE AGRO BASED INDIAN PULP & PAPER INDUSTRY

INTRODUCTION The Indian Pulp & Paper Industry is more than a century old industry . The first paper mill was established in the year 1832 and today the number has grown to more than 400 in the organised sector which are located through out the country . In addition there are another 500 pulp & paper mills in the unorganized sector. The Indian pulp & paper industry can be categorised into large, medium and small depending on the scale of operation as indicated in Table -I Most of the large mills established initially were based on woody raw materials.     

Salt tolerance conferred by over-expression of OsNHX1 gene in Poplar 84K

OsNHX1 gene (Na^＋/H^＋ antiporter gene of Oryza sativa L.) was introduced into Poplar 84K with Agrobacterium tumefaciens-mediated transformation. PCR, Southern and Northern blot analysis showed that OsNHX1 gene was incorporated successfully into the genome of Poplar 84K and expressed in these transgenic plants. Salt tolerance test showed that three lines of transgenic plants grew normally in the presence of 2OO mmol/L NaCI, while the Na^＋ content in the leaves of the transgenic plants grown at 2OO mmol/L NaCl was significantly higher than that in plants grown at 0mmol/L NaCI. The osmotic potential in the transgenic plants with high salinity treatment was lower than that of control plants. Our results demonstrate the potential use of these transgenic plants for agricultural use in saline soils.     

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.     

Application of Chitosan Flocculant to Conditioning Sludge

The dewaterability of activated sludge conditioned by chitosan flocculant was studied. The effects of chitosan characteristics such as molecular weight, degree of deacetylation, and dose on the dewaterability were investigated. The sludge dewaterability is evaluated in terms of specific resistance to filtration, residual turbidity of supernatant, moisture content of cake, and settling rate. Sludge dehydrating behaviors conditioned with CTS, PAM and PAC flocculants were compared. The conditioning was also carried out with dual flocculants in two stages. It is found that the sludge conditioned with CTS has better dewaterability than that with PAC. The optimum conditions with chitosan are: dose 0.8 - 1.2 g per 100 g dry cake, molecular weight 300,000, and degree of deacetylation 70%. The conditioning in two stages with dual flocculants is found to be more effective than that with single flocculant.     

Settling characteristics of ultrafine iron ore slimes

The slurry settling characteristics are the most important to design a thickener in process industries. In this work, the iron ore slurry from the screw classifier overflow was used for the settling study. It was observed that the original slurry exhibited a low settling velocity and a turbid supernatant during the settling process. Commercial flocculating agents with anionic, cationic, and nonionic characters were used to improve the settling behavior of suspensions, which were added into the slurry at different ranges of slurry pH values, respectively. The settling results show that the use of flocculants increase the settling rate by several times. Compared with the cationic and nonionic flocculants, the anionic flocculant is more effective in enhancing the slurry settling rate. The small dose of the anionic flocculant is found to be more effective, but the other flocculants are less effective even at higher dosages. The simulation of an industrial thickener was carried out based on the laboratory settling data, and the appropriate design and selection parameters of the industrial thickener were estimated.     

Obtaining High Pest-resistant Tobacco Plants Carrying B.t. insecticidal Gene

To increase the expression level of CryIA(c) gene in transgenic plants, a plant expression vector pBinMoBc carrying the CryIA(c) gene under control of chimeric OM promoter and Ω factor was constructed. As a control, pBinoBc carrying the CryIA(c) gene with the CaMV 35S promoter was also constructed. The vectors were transferred into tobacco plants respectively via Agrobacterium-mediated transformation. ELISA assay showed that the expression level of the CryIA(c) gene in pBinMoBc transgenic tobacco plants was 2.44-times that in pBinoBc transgenic tobacco plants, and it could be up to 0.255% of total soluble proteins. Bioassay showed that pBinMoBc transgenic tobacco plants had more notable insecticidal effect than pBinoBc transgenic tobacco plants. The above results showed that the chimeric OM promoter was a stronger promoter than CaMV 35S promoter that was widely used in plant genetic engineering, and this is very useful in pest-resistant plant genetic engineering.     

Impact of gamma radiation and salinity on growth and K^＋/Na^＋ balance in two populations of Medicago sativa （L.） cultivar Gabbs

The perennial Medicago sativa cv. Gabes is widely grown on saline soils in Tunisian oases. Morphological and physiological analyses of two populations （Mareth and Gannouch） were conducted in order to study the effect of salinity and gamma radiation （350 Gy） interaction on two populations of this species. It has been shown that the two irradiated populations of Medicago sativa are fairly tolerant to salt at growth phase compared to the non-irradiated. Exposure to gamma irradiation （350 Gy）, alone or in combination with salt stress, increased significantly （p 〈 0.001） shoot number, stem height and chlorophyll b pigment especially for the Gannouch population, while no change occurred for the Mareth population. The presence of salt to 9 g/1 affected significantly the root biomass and induced a reduction of shoot development of both control and irradiated alfalfa populations. For all treatments, plants of two populations remained able to produce and to allocate dry matter to the different organs. The survey of Na^＋/K^＋ ratio showed that the growth of the aerial organs of two non-irradiated populations was at least determined by a selectivity in favour of the K^＋ ions （r^2 = 0.97 and r^2 = 0.59 for Mareth and Gannouch non-irradiated populations, respectively）. However, the rather weak correlation detected for the irradiated populations, particularly for the irradiated Gannouch, seems to be the consequence of the effect of irradiation that improved potassium availability, essential element for growth and development. Results also showed that the two irradiated populations, especially the Mareth, accumulated Na^＋ ions in its photosynthetic organs. This accumulation was associated with an improvement of foliar water content at a level of salinity around 5 g/1. Such a mechanism reflects probably an inclusive behaviour of the plants and a good aptitude to use the dominant ions （Na^＋） for the osmotic adjustment. However, the non-irradiated populations are unable to adjust their internal os     

SRO1 regulates heavy metal mercury stress response in Arabidopsis thaliana

Heavy metals in the environment are harmful limiting factors for the normal growth and development of plants. Here, we isolated and identified an Arabidopsis thaliana T-DNA insertion mutant, named srol-1, which showed a hyper-sensitive response to HgCl2. The SRO1 protein contains a WWE domain that mediates proteinprotein interactions. Under HgCl2 treatment, when compared with the wild-type plants, the growth of srol-1 was repressed dramatically and the number of true leaves was reduced and etiolated. The electrolyte leakage rates showed that cell membrane integrity in srol-1 was damaged more severely than in the wild type. DAB （3,5-diaminobenzidine） staining and confocal microscopy showed that Hg2＋ stress induced more hydrogen peroxide accumulation in srol-1 than in the wild type. The qRT-PCR results indicated that the expression of some abiotic stress-induced genes, such as L-ascorbate peroxidase （APX1）, was reduced under oxidative or Hg2＋ stress. Transgenic plants containing a GFP：：SRO1 fusion protein showed that SRO1 was localized in the nucleus of the cells. SRO1 was shown to be expressed in various tissues, and was most highly expressed in the vigorous tissues. Our results suggest thatSRO1 may play an important role in the stress response of A. thaliana to heavy metals.     

Accumulation and distribution of arsenic and cadmium by tea plants

It is important to research the rules about accumulation and distribution of arsenic and cadmium by tea plants, which will give us some scientific ideas about how to control the contents of arsenic and cadmium in tea. In this study, by field investigation and pot trial, we found that mobility of arsenic and cadmium in tea plants was low. Most arsenic and cadmium absorbed were fixed in feeding roots and only small amount was transported to the above-ground parts. Distribution of arsenic and cadmium, based on their concentrations of unit dry matter, in tea plants grown on un-contaminated soil was in the order： feeding roots〉stems=main roots〉old leaves〉young leaves. When tea plants were grown on polluted soils simulated by adding salts of these two metals, feeding roots possibly acted as a buffer and defense, and arsenic and cadmium were transported less to the aboveground parts. The concentration of cadmium in soil significantly and negatively correlated with chlorophyll content, photosynthetic rate, transpiration rate and biomass production of tea plants.     

Evaluation of Cadmium Bioavailability in Soils Using Diffusive Gradients in Thin Film Technique and Traditional Methods

The developed technique of diffusive gradients in thin film( DGT) has been suggested as a promising tool for evaluation of cadmium( Cd) availability in soils,but there exists considerable debate on its suitability. In this study,Cd bioavailability in soils was systemically investigated by comparing this technique with seven traditional methods, including soil solution concentration and six commonly used extraction methods( HAc,EDTA,Na c,NH4Ac,CaCl2 and MgCl2). Two typical plants( wheat and maize) were examined for Cd uptake. Maize was more sensitive to increasing exposure to Cd in soils than wheat when the added amounts of Cd ranged from 3. 5 to 5. 0 mg · kg-1,accompanied by the significant decreases of shoot and root biomasses. Cd concentrations in shoots and roots of two plants increased continuously with increasing levels of Cd in soils. Cd uptake was higher in wheat than that in maize. The bioavailable concentrations of Cd measured by all methods increased nearly linearly with increasing addition of Cd in soils. Positive correlations were observed between various bioavailable indicators of Cd in soils and Cd concentrations in two plants. The extractable Cd by six chemical extractants was considerably higher for maize than for wheat,while the DGT-measured and soil solution concentrations of Cd were lower for maize than for wheat,following the same trend as plant Cd uptake. The results imply that DGT measurement can effectively predict the bioavailable levels of Cd in soil solutions and that it is an ideal tool for prediction of Cd bioavailability in soils.     

A five-fold pig bacterial artificial chromosome library: a resource for positional cloning and physical mapping

A pig BAC library was constructed with genomic DNA from a male Erhualian pig. After partial digestion with Hind III or BamH I the fragments obtained were cloned into the pBeloBAC11 vector. The library consists of 184320 clones which stored in 480 pieces 384-well plates (20 plates per superpool). A two-step 4-dimension PCR screening system was established to screen the positive clones. An average insert size of 128 kb was estimated from 105 randomly isolated clones, which indicates that the library is more than five times of genomic coverage. For the demonstration of the probability to pick out any unique genes or DNA markers from the library, 10 single-copy genes were screened out and the positive clones were yielded between 1 and 8 with an average of 3.6. Positive superpools were obtained for 32 microsatellite markers selected from different regions of pig genome. The number of positive superpools for each marker varies from 1 to 9 with an average of 4.78. This BAC library provides an additional resource for pig physical mapping and gene identification.     

Host selection of Helicoverpa armigera and H. assulta and its inheritance

The difference in host selection between the polyphagous Helicoverpa armigera (Hübner) and the oligophagous H. assulta Quenée was examined, with cotton (Gossypium hirsutum), tomato (Lycopersicon esculentum) (hosts of H. armigera), tobacco (Nicotiana tobaccum) (host of both H. armigera and H. assulta), and bush redpepper (Capsicum frutescens) (host of H. assulta) as testing plants. A multiple-choice test was used with caged plant cuttings for adult oviposition and with leaf discs for larval feeding. A no-choice test was run for evaluating larval growth rate. The results indicated that the relationship between larval performance and adult preference of H. assulta was more conspicuous than that of H. armigera. Reciprocal hybridization between H. armigera and H. assulta followed by backcrossing of the hybrids (F1) with H. armigera was also carried out for genetic study on host selection of these two insect species. A two-choice test with cotton and bush redpepper leaf discs showed that H. armigera larvae preferred to feed on cotton, and H. assulta larvae to bush redpepper; feeding preferences of the two F1 lines were intermediate between those of their parents, but close to that of their female parent; preference indexes of backcross lines also showed that both maternal factor and chromosomal inheritance were involved in feeding selection of two Helicoverpa species.     

Behavioral and olfactory responses of grasshopper hatchlings,<Emphasis Type="Italic">Melanoplus sanguinipes</Emphasis>, to plant odours and volatile compounds

Behavior and olfactory responses of grasshopper hatchlings, Melanoplus sanguinipes (F.), to odours from plant foliage and volatile compounds were tested using a glass Y-tube olfactometer and electroantennogram (EAG)techniques respectively. In single choice trials, newly hatched hoppers were much more sensitive to the odour from intact leaves and chopped foliage of ryegrass and wheat than other plants. Chopped sorghum leaves, but not stem-cut sorghum,were also significantly attractive. The orientation responses of grasshopper hatchlings to these plants were highly consistent with those of last instar hoppers and adults. When ryegrass was employed as the control, the odour from stem-cut alfalfa was more attractive. There was no significant difference in hopper orientation responses to the odours from chopped seedlings of sorghum, alfalfa, wheat or ryegrass.However, significantly more hoppers preferred the chopped ryegrass control to chopped Louisanna sage. Measurement of the EAG response of first instar hoppers to these plant odours showed that the odour of Louisanna sage elicited the greatest response amplitudes. In olfactory tests using different volatile components, Z-3-hexenol, E-3-hexenol,Z-hex-3-enyl acetate, E-2-hexenal and hexenal gave greater EAG responses than geraniol and 1-octen-3-ol. These results are also consistent with comparable data from adults. Newly hatched grasshoppers had similar EAG response profiles to plant materials and chemicals to those of adults, although the absolute EAG values of young hoppers were much lower than those of adults. Therefore, newly hatched hoppers were able to distinguish plants from an air control, and even host plants from non-host plants, and the feeding experience of hoppers probably has little influence on their subsequent ability as adults to identify and locate food plants.     

Characterization of variation induced by low-energy N+ and cloning of differentially expressed cDNA of a mutant in Arabidopsis thaliana

Using Arabidopsis thaliana as experimental materials, the variations induced by low-energy N+ have been investigated. Germination rate of the treated seeds is lower than that of the control, and it decreases with the intensification of the radiation. The phenotypic variations have been observed in M2 plants irradiated with higher doses, such as chlorisis, semilethality, plant morphology, and changes of blooming habit and fertility. In random amplified polymorphic DNA (RAPD) analysis on M2 seedlings, some differences including band deletions or additions are found in treated plants compared to the control and the differences are associated with the radiation doses. One of the M1 plants from the seeds irradiated with the dose of 80×1015 N+/cm2 is a dwarf variant. Its stable M6 generation, mutant T80II, is used to construct subtractive cDNA library and to clone differentially expressed cDNA. A 721 bp cDNA fragment is partly homologous with GRF7 gene.     

Design and implementation of binary tree data structure based on DNA computing

The designing,encodings and an instance of simulation of a binary tree for DNA computer were proposed,which utilizes the method of biology to complete inserting and deleting of the binary tree. Firstly,DNA encodings for storage and all elements of the binary tree were completely given out. Then, the implementations of all biooperations in DNA computer were described. Finally, to prove the feasibility of this method, an actual binary tree with detailed nucleotide encodings was introduced. The process of an algorithm implemented on this binary tree was demonstrated. Based on this method, more other data structures in DNA computer can be developed.     

Genetic diversity and construction of core collection in Chinese wheat genetic resources

Genetic diversity among 5029 accessions representing a proposed Chinese wheat core collection was analyzed using 78 pairs of fluorescent microsatellite （SSR） primers mapped to 21 chromosomes. A stepwise hierarchical sampling strategy with priority based on 4×10^5 SSR data-points was used to construct a core collection from the 23090 initial collections. The core collection consisted of 1160 accessions, including 762 landraces, 348 modern varieties and 50 introduced varieties. The core accounts for 23.1% of the 5029 candidate core accessions and 5% of the 23090 initial collections, but retains 94.9% of alleles from the candidate collections and captures 91.5% of the genetic variation in the initial collections. These data indicate that it is possible to maintain genetic diversity in a core collection while retaining fewer accessions than the accepted standard, i.e., 10% of the initial collections captured more than 70% of their genetic diversity. Estimated genetic representation of the core constructed by preferred sampling （91.5%） is much higher than that by random sampling （79.8%）. Both mean genetic richness and genetic diversity indices of the landraces were higher than those of the modern varieties in the core. Structure and principal coordinate analysis revealed that the landraces and the modern varieties were two relatively independent subpopulaUons. Strong genetic differentiation associated with ecological environments has occurred in the landraces, but was relatively weak in the modern culUvars. In addition, a mini-core collection was constructed, which consisted of 231 accessions with an estimated 70% representation of the genetic variation from the initial collections. The mini-core has been distributed to various research and breeding institutes for detailed phenotyping and breeding of genetic introgression lines.