Yang ChuanPing

Effect of salt stress on activity of superoxide dismutase (SOD) in Ulmus pumila L.

AbstractsThe injury tolerance of cell plasma membrane and the correlative enzymes activities of plasma-membrane protection system in the Ulmus pumila leaves treated by nine concentrations (0.3%, 0.6%, 0.9%, 1.2%, 1.5%, 1.8%, 2.1%, 2.4%, 3.0%) of Na2CO3 and NaHCO3 mixtures were studied in a greenhouse of Northeast Forestry University, Harbin, China. The rate of electrolyte leakage (REL) and SOD (Superoxide dismutase) activity in leaves of different samples were determined. Results showed that the REL in leaves of U. pumila presented a slowly increasing trend at the salt concentrations less than 1.5%, which indicated that cell plasma membrane of U. pumila leaves had rather strong resistance to the injury of salt ion, and had a significant increase at the salt concentrations more than 1.5%. The SOD activities in leaves of U. pumila presented an increased trend at salt concentrations less than 1.5%, the growth of seedlings did not decline, and tress and leaves had no symptom of injury, while the salt concentrations exceeded 1.5%, SOD activities sharply decreased and REL increased promptly.

DNA extraction of birch leaves by improved CTAB method and optimization of its ISSR system

The basic method of DNA extraction (CTAB) was improved as the multi-times STE-CTAB extraction method and used to extract the DNA of birch leaved in this experiment. Results showed that the improved method is suitable not only for genomic DNA extraction of birch but also for that of other plants. The purity of genomic DNA extracted by the multi-times STE-CTAB extraction method is higher than that by one time STE-CTAB method, and it does not need the process of RNase. The factors of influencing ISSR system were explored based on the genomic DNA of birch extracted by the two methods. The optimal conditions for ISSR system were determined as follows: Mg2+ concentration is 1.5–3.0 mmol·L−1, dNTP concentration 0.10–0.25 mmol·L−1, the quantity of Taq polymerase 0.5–2.0 U, template DNA 30–100 ng, and the concentration of primer is 0.2–0.4 μmol·L−1, and the reaction program was as: initial denaturation for 5 min at 94°C, 30 cycles of denaturation for 30 s at 94°C, annealing for 30 s at 51 °C, extension for 30 s at 72°C, and a final 7 min extension at 72 °C.

Differential Expression of Endogenous Ferritin Genes and Iron Homeostasis Alteration in Transgenic Tobacco Overexpressing Soybean Ferritin Gene

For studying the effects of endogenous ferritin gene expressions (NtFer1, GenBank accession number ay083924; and NtFer2, GenBank accession number ay141105) on the iron homeostasis in transgenic tobacco (Nicotiana tabacum L.) plants expressing soybean (Glycine max Merr) ferritin gene (SoyFer1, GenBank accession number m64337), the transgenic tobacco has been produced by placing soybean ferritin cDNA cassette under the control of the CaMV 35S promoter. The exogenous gene expression was examined by both Northern- and Western-blot analyses. Comparison of endogenous ferritin gene expressions between nontransformant and transgenic tobacco plants showed that the expression of NtFer1 was increased in the leaves of transgenic tobacco plants, whereas the NtFer2 expression was unchanged. The iron concentration in the leaves of transgenic tobacco plants was about 1.5-folds higher than that in nontransformant. Enhanced growth of transgenic tobacco was observed at the early development stages, resulting in plant height and fresh weights significantly greater than those in the nontransformant. These results demonstrated that exogenous ferritin expression induced increased expression of at least one of the endogenous ferritin genes in transgenic tobacco plants by enhancing the ferric chelate reductase activity and iron transport ability of the root, and improved the rate of photosynthesis.

Construction and Analysis of a Subtracted cDNA Library of Betula platyphylla Female Inflorescence

Female inflorescence ofBetula platyphylla was sampled at an interval of each two days to analyze the background of gene expression in floral phase. On the basis of SMART strategy, the driver cDNA was obtained from total RNA of the last sample and the tester cDNA was from that of the others by RT-PCR which were subsequently used to construct a subtracted cDNA library. The result of the ESTs (expression sequence tags) blastX showed that the genes in the subtracted cDNA library could be mainly clustered into 5 groups related to metabolism, transportation and signal transduction, cell cycle, stress response, and regulation. The relationship between gene expression and development was also discussed.

Cloning and stress tolerance analysis of an LbGRP gene in Limonium bicolor

Glycine-rich RNA-binding proteins (GRP) are involved in post-transcriptional regulation, and play important roles in plant growth, development and stress response. In the present study, the full-length cDNA of a GRP gene (LbGRP, GenBank accession number: GQ398238) was cloned from a cDNA library of Limonium bicolor. To investigate the stress tolerance of LbGRP, the recombinant plasmid pYES2-LbGRP was constructed by inserting the LbGRP gene into the yeast expression vector pYES2. The recombinant plasmid pYES2-LbGRP was transformed into yeast Saccharomyces cerevisiae INVSc1, and the INVSc1 strain transformed with empty pYES2 was used as the control. The recombinant yeast INVSc1 harboring LbGRP (pYES2-LbGRP) and the control INVSc1 harboring empty pYES2 were treated with NaCl, KCl, NaHCO3, Na2CO3, drought and frezzing, respectively, and their survial rates were compared under these stress conditions. The results showed that the survival rates of the recombinant yeast INVScl (pYES2-LbGRP) were higher than that of the control strain under these stress conditions, indicating that the LbGRP is tolerant to NaCl, KCl, NaHCO3, Na2CO3, drought and frezzing stresses. These results suggested that LbGRP plays a role in stress tolerance of L. bicolor.Glycine-rich RNA-binding proteins (GRP) are involved in post-transcriptional regulation, and play important roles in plant growth, development and stress response. In the present study, the full-length cDNA of a GRP gene (LbGRP, GenBank accession number: GQ398238) was cloned from a cDNA library of Limonium bicolor. To investigate the stress tolerance of LbGRP, the recombinant plasmid pYES2-LbGRP was constructed by inserting the LbGRP gene into the yeast expression vector pYES2. The recombinant plasmid pYES2-LbGRP was transformed into yeast Saccharomyces cerevisiae INVSc1, and the INVSc1 strain transformed with empty pYES2 was used as the control. The recombinant yeast INVSc1 harboring LbGRP (pYES2-LbGRP) and the control INVSc1 harboring empty pYES2 were treated with NaCl, KCl, Na- HCO3, Na2CO3, drought and frezzing, respectively, and their survial rates were compared under these stress conditions. The results showed that the survival rates of the recombinant yeast INVScl (pYES2-LbGRP) were higher than that of the control strain under these stress conditions, indicating that the LbGRP is tolerant to NaCl, KCl, NaHCO3, Na2CO3, drought and frezzing stresses. These results suggested that LbGRP plays a role in stress tolerance of L. bicolor.

Construction and analysis of a cDNA library from yellow-fruit ginseng ( Panax ginseng C.A.Meyer.) leaf tissue

The total RNA was isolated from yellow-fruit ginseng (Panax ginseng C.A. Meyer) leaf tissue. A cDNA library of panax ginseng leaves was constructed by using pDNR-LIB vector according to the SMART cDNA library construction kit protocol. We obtained 378 high quality sequences (GenBank accession number: ES672876-ES673253). ESTs were annotated, analyzed by BlastX and functional classified based on gene ontology, the results showed that 221 ESTs showed significant similarities to gene sequences in Nr database and were known genes, 21 ESTs were non-significance and unknown function genes, and 136 ESTs were considered novel genes. Most of the ESTs appeared to be related to physiological and cellular processes.

Cloning and sequence analysis of gene encoding plasma aquaporin of Tamarix albiflonum

Plant aquaporins are water-selected-channels in plants and are involved in seed germination, cell elongation, stoma movement, fertilization and so on. Some plant aquaporins also play an important role in drought stress response. In this paper, the gene encoding the Tamarix albiflonum Aquaporin (AQP) was amplified by 5′rapid amplification of cDNA end (RACE) on the basis of the sequence information obtained from the expressed sequence tag of the subtractive hybridization library constructed under PEG6000 stress. The cDNA of the T. albiflonum AQP gene is 1,043 bp long, encoding a protein of 287 amino acids with a predicted molecular mass of 30.9 kDa, has 6 transmembrane regions, and possessing the major intrinsic protein (MIP) family signal consensus sequence SGXHXNPAVT and the higher plant plasma membrane intrinsic protein (PIP) highly conservative sequence GGGANXXXXGY and TGI/TNPARSL /FGAA I/VI/VF/YN. A comparative molecular analysis of the nucleotide sequence in National Center for Biotechnology Information (NCBI) databases showed that it shared 95% homology with the gene of Arabidopsis thaliana (MIP-C), with a theoretical isoelectric point 8.84.

Study on the provenance test of Dahurian Larch selection of best provenance

In this paper, it is studied that the growth characters of the progeny of 16 provenance oflarix gmelinii within the all 13 experimental sites by means of the analysis of variance, provenance stability, productivity index, rank correlation, synthetic index selection and PCA.The provenances were divided into three patterns: The first type have low and steady productivity; The second type have high but unsteady productivity; The third type is between the productivity of the first and second type. Through the analysis on the feasibility and the reliability of the provenance early selection, the best provenances which lie in the southeast of the Xiaoxingan Mt. are regarded as the best and suitable for a large afforestation areas ofLarix gmelinii. Then in the north of Xingan Mt., the local or the northwest provenances of the Xiaoxingan Mt. should be used. The genetic gain was calculated on the basis of the provenance heritability and the utilization of the best provenance were evaluated in this paper.

Construction and analysis of a forward and reverse subtractive cDNA library from leaves and stem of Polygonum sibiricum Laxm.under salt stress

Using cDNAs prepared from the leaves and stems of Polygonum sibiricum Laxm. treated with NaHCO3 stress for 48 h as testers and cDNAs from unstressed P. sibiricum leaves and stems as drivers library, suppression subtractive hybridization (SSH) was employed to construct a cDNA subtracted library, which contained 2 282 valid sequences including 598 ESTs in the stems forward SSH library and 490 ESTs in the stem reverse SSH library, 627 ESTs in the leaf forward SSH library and 567 in the leaf reverse SSH library. According to the functional catalogue of MIPs and the comparison of the reverse and forward SSH libraries of the stem and leaf, the responses to NaHCO3 stress were different between leaf and stem, except for the same trend in cell rescue defense and transport facilitation. The trend in the metabolism, energy, photosynthesis, protein synthesis, transcription, and signal transduction was opposite. RT-PCR analysis demonstrated that the expression of 12 putative stress related genes in the NaHCO3-treated leaves and stems was different from that in the untreated leaves and stems. This indicated that different mechanisms might be responsible for reactions of leaf and stem in P. sibiricum. The results from this study are useful in understanding the molecular mechanism of saline-alkali tolerance in P. sibiricum.

Molecular characterization and fermentative hydrogen production of a wild anaerobe in clostridium genus

Anaerobic process of biohydrogen production is developed in this paper. The isolation and identification of high efficient biohydrogen production anaerobic bacteria are the important foundations for the fermented biohydrogen production process by anaerobic digesting organic wastewater. Taking the physiological and biochemical traits, the morphological characteristics and 16S rDNA sequence into consideration, the isolate Rennanqilyf33 is a new species.