Harmonized biochemical modification of cell walls to get permission for entrance of Azospirillum sp. to rice roots.

Biological nitrogen-fixation is important in increasing crop efficiency. Azospirillum is a nitrogen-fixing microorganism that naturally coexists with grasses roots. The present study was undertaken to clarify the role of rice root cell walls in the acceptance of two Azospirillum species, alone or in combination with indole-3-acetic acid (IAA) and gibberellic acid (GA3) treatments. Rice seedlings were grown in Yoshida solution for 21 days and then inoculated with A. brasilense and A. irakens in the presence of 0, 0.57, and 1.14 mM of IAA or 0, 0.29, and 0.58 mM GA3 or a combination of 1.14 mM of IAA and 0.58 mM of GA3. The results showed that the amount of hydrogen peroxide, lipid peroxidation, total nitrogen and activity of ferulic acid peroxidase, NADPH oxidase, nitrate reductase, pectin methyl esterase, cellulase, mannanase, xylanase and pectinase were significantly increased in inoculated samples treated with or without phytohormones. The highest activity of these enzymes was observed in A. brasilense- inoculated rice roots in auxin+gibberellin treatment. In the latter, the activity of phenylalanine ammonia lyase and wall ferulic acid peroxidase enzymes, the content of cell wall polysaccharide, lignin, and total phenolic compounds were the least, compared to controls and also with those samples which were inoculated with A. irakens. The results indicate an active role of the wall and its enzymes in allowing bacteria to enter the roots. Understanding this mechanism can improve the methods of inoculating bacteria into plants and increase crop efficiency, which will result in reduced use of chemical fertilizers and their destructive environmental effects.

met1 DNA Methyltransferase Controls TERT Gene Expression: A New Insight to The Role of Telomerase in Development.

OBJECTIVE: DNA methylation systems are essential for proper embryo development. Methylation defects lead to developmental abnormalities. Furthermore, changes in telomerase gene expression can affect stability of chromosomes and produces abnormal growth. Therefore, defects in both methylation and telomerase gene expression can lead to developmental abnormalities. We hypothesized that mutation in the methylation systems may induce developmental abnormalities through changing telomerase gene expression. MATERIALS AND METHODS: In this experimental study, we used Arabidopsis thaliana (At) as a developmental model. DNA was extracted from seedlings leaves. The grown plants were screened using polymerase chain reaction (PCR) reactions. Total RNA was isolated from the mature leaves, stems and flowers of wild type and met1 mutants. For gene expression analysis, cDNA was synthesized and then quantitative reverse transcription PCR (qRT-PCR) was performed. RESULTS: Telomerase gene expression level in homozygous met1 mutant plants showed ~14 fold increase compared to normal plants. Furthermore, TERT expression in met1 heterozygous was~ 2 fold higher than the wild type plants. CONCLUSION: Our results suggested that TERT is a methyltransferase-regulated gene which may be involved in developmental abnormities causing by mutation in met1 methyltransferase system.

Silicon increases cell wall thickening and lignification in rice (Oryza sativa) root tip under excess Fe nutrition.

Iron (Fe) as a micronutrients and silicon (Si) as a cell wall element are important in plant cell wall extension and integrity. While the interaction of exogenous Si and excess Fe on root cell wall modifications is known, the effects of these nutritional parameters on the spatial changes in the activities of genes and/or enzymes involved in the lignification of root cell walls are not well studied. Thus, these parameters were investigated in the root apical part (AP) and basal part (BP) of rice (Oryza sativa L.) plants supplied with and without Si (1.5 mM) under normal (10 mg/L) and excess Fe (150 mg/L) nutrition for 7 days. Beside growth retardation, excess Fe increased the activities of phenylalanine ammonia lyase (PAL), superoxide dismutase and NADPH-oxidase and PAL and cell wall peroxidase (POD) genes expression, along with the increased phenols and H2O2 contents in the root AP. Furthermore, the increased thickening of endodermal, exodermal and metaxylem cell walls in the root AP by excess Fe was attributed to the enhanced POD activity. POD expression, endodermal and exodermal cell wall thickenings were not affected by excess Fe in the root BP. Si application under excess Fe exaggerated the effects of excess Fe on root cell wall thickening, increased POD activity but reduced H2O2 content in the root AP. Thus, Si application under excess Fe nutrition promotes earlier initiation of lignin polymerization closer to and toward the root tip and hence restricts the entry of excess Fe into the plant.

Redox metabolism and cell wall modifications as global and local targets respectively, of cyanide induced dormancy release of walnut kernels.

The HCN-induced seed dormancy release necessitates alterations in reactive oxygen species (ROS) metabolism and radicle cell wall loosening. Little is known about the interaction of ROS metabolism with cell wall hydrolytic enzymes during HCN-induced seed dormancy release. Thus dormant walnut (Juglans regia L.) kernels were exposed to HCN (4 h) and studied for redox metabolism and cell wall-modifying enzymes during 10 days of incubation (DI) i.e. before radicle emergence. HCN increased ROS especially in the embryonic axes (EAs) but decreased ROS-generating NADPH oxidase and ROS scavenging superoxide dismutase (SOD) and peroxidase (POX) with no effects on catalase (CAT), ascorbate peroxidase (APX) and cell wall-modifying enzymes activities in short term up to 2 DI. In long term roughly from 4 DI onwards, HCN-exposed EA displayed greater superoxide anions and enhanced activities of POX, APX, NADPH oxidase, cell wall peroxidase (CW-POX), ß- 1, 4-D glucanase, mannanase, polygacturonase and xylanase. Meanwhile HCN increased greater expression of POX and mannanase isoforms as revealed by in-gel activity assay. Except for higher activities of CAT, POX and APX, cotyledonary activities of CW-POX, mannanase and polygacturonase and to some extent ß- 1, 4-D glucanase remained unaffected by HCN. Thus short term ROS accumulation in HCN-treated EA is due to declined SOD and POX activities. In long term the enhanced activities of both NADPH oxidase: CW-POX couple and cell wall-modifying enzymes in EA bring about wall loosening in preparation for radicle emergence. Evidences for the simultaneous operation of both mechanisms are provided in walnut EAs during dormancy release.

The Impacts of TRR14-Overexpression on Arabidopsis thaliana Growth and Photosynthetic Parameters.

BACKGROUND: TRR14 protein is a small protein, a member of a multigene family in Arabidopsis which was found as the fi rst protein during screening seedlings for their resistant to the trehalose sugar. OBJECTIVES: A number of TRR14-overexpressing plants were subjected to the characterization in the present research, among which, the associated morphological features and changes accompany growth pattern and photosynthesis related parameters. MATERIALS AND METHODS: TRR14 gene was isolated from Arabidopsis Thaliana and cloned into the pBin-35S vector. Recombinant vector was transferred to the Arabidopsis (Col-0) via Agrobacterium tumefaciens using the Floral Dipping method. Seeds from the TRR14 overexpressed (TRR14) and the Col-0 wild-type (WT) plants were shown on soil under long day conditions. Several measurements were then performed including determination of the fresh and dry weights, leaf area, chlorophyll a and b (Chl a and Chl b) content, Chl a/b ratio, total chlorophyll and carotenoids content, soluble and insoluble sugars content, total and soluble protein content, the Hill reaction rate, chlorophyll fl uorescence, as well as photorespiration rate. Meanwhile, the chloroplastic proteins were investigated by SDS-PAGE analysis. RESULTS: TRR14 plants showed a signifi cant increase in fresh and dry weights, leaf area, and total and soluble protein content along with a signifi cant decrease in the insoluble sugar contents was observed in comparison to the WT plants. Chl a, Chl b, total chlorophyll content, Chl a/b ratio, carotenoids content, Hill reaction rate, and chlorophyll fl uorescence didn't show a signifi cant diff erence between TRR14 and WT plants. The SDS-PAGE gel electrophoresis of the chloroplastic proteins showed a thick band with a molecular mass of 25 kDa in TRR14-overexpressed plants, compared to the WT plants. Remarkably, photorespiration rate was decreased in TRR14 plants compared to WT plants. CONCLUSION: The increased biomass of TRR14 transformed plants might be due to its ability in reducing photorespiration through concentrating CO2 in the leaf's intercellular spaces.

The dynamics of the bacterial communities developed in maize silage.

Ensilage provides an effective means of conserving summer-grown green forage to supply as winter feed to ruminants. The fermentation process involved in the ensilage process relies on lactic acid bacteria (LAB). Here, 16S ribosomal DNA amplicon pyrosequencing was used to follow the dynamic behaviour of the LAB community during the ensilage of maize biomass, with a view to identify the key species involved in the process. The biomass used for ensilage was a single-cross maize hybrid, harvested at the milk-line stage. The crop was grown at three contrasting locations. Aspects of the physico-chemical composition of the material and the LAB species present were sampled at 0, 3, 6, 14, 21 and 32 days after ensilage was initiated. In all three cases, members of the Leuconostocaceae family dominated the epiphytic bacterial community, notably Leuconostoc and Weissella, but some variation was noted in the abundance of certain Leuconostocaceae and Lactobacillaceae species, as well as that of some Acetobacteraceae, Enterobacteriaceae and Moraxellaceae species. The constellation of the microbiome which developed during the ensilage process differed markedly from that of the epiphytic one, with Lactobacillaceae, particularly Lactobacillus and Pediococcus spp. dominating. The abundance of heterofermentative Leuconostocaceae spp. in the epiphytic community and the extent of the transition from hetero- to homo-fermentation during the initial ensilage period are important factors in determining silage quality.

Short versus long term effects of cyanide on sugar metabolism and transport in dormant walnut kernels.

Tree seed dormancy release by cold stratification accompanies with the embryo increased gluconeogenesis competence. Cyanide also breaks seed dormancy however, integrated information about its effects on carbon metabolism is lacking. Accordingly, the impacts of HCN on germination, lipid gluconeogenesis and sugar transport capacity of walnut (Juglans regia L.) kernels were investigated during 10-days period prior to radicle protrusion. HCN increased walnut kernel germination and within four days of kernel incubation, hastened the decline of starch, reducing and non-reducing sugars and led to greater activities of alkaline invertase and glucose-6-phosphate dehydrogenase. From four days of kernel incubation onwards, starch and non-reducing sugars accumulated only in the HCN treated axes. Cyanide also increased the activities of phosphoenolpyruvate carboxykinase and glyoxysomal succinate oxidase and led to greater acid invertase activity during the aforementioned period. The expressions of both sucrose transporter (JrSUT1) and H+-ATPase (JrAHA1) genes especially in cotyledons and H+-ATPase activity in kernels were significantly enhanced by exposure to cyanide. Thus in short-term HCN led to prevalence of carbohydrate catabolic events such as oxidative pentose phosphate pathway and possibly glycolysis in dormant walnut kernels. Long-term effects however, are increased gluconeogenesis and enhanced sugar transport capacity of kernels as a prerequisite for germination.

Trehalose mediated growth inhibition of Arabidopsis seedlings is due to trehalose-6-phosphate accumulation.

Trehalose-6-phosphate (T6P) is required for carbon utilization during Arabidopsis development, and its absence is embryo lethal. Here we show that T6P accumulation inhibits seedling growth. Wild-type seedlings grown on 100 mm trehalose rapidly accumulate T6P and stop growing, but seedlings expressing Escherichia coli trehalose phosphate hydrolase develop normally on such medium. T6P accumulation likely results from much-reduced T6P dephosphorylation when trehalose levels are high. Metabolizable sugars added to trehalose medium rescue T6P inhibition of growth. In addition, Suc feeding leads to a progressive increase in T6P concentrations, suggesting that T6P control over carbon utilization is related to available carbon for growth. Expression analysis of genes from the Arabidopsis trehalose metabolism further supports this: Suc rapidly induces expression of trehalose phosphate synthase homolog AtTPS5 to high levels. In contrast, T6P accumulation after feeding trehalose in the absence of available carbon induces repression of genes encoding T6P synthases and expression of T6P phosphatases. To identify processes controlled by T6P, we clustered expression profile data from seedlings with altered T6P content. T6P levels correlate with expression of a specific set of genes, including the S6 ribosomal kinase ATPK19, independently of carbon status. Interestingly, Suc addition represses 15 of these genes, one of which is AtKIN11, encoding a Sucrose Non Fermenting 1 (SNF1)-related kinase known to play a role in Suc utilization.