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Biochemistry And Molecular Biology Of Plants

Salicylic acid-induced nitric oxide enhances arsenic toxicity tolerance in maize plants by upregulating the ascorbate-glutathione cycle and glyoxalase system.

Glyphosate, the most commonly used herbicide in the world, controls a wide range of plant species, mainly because plants have little capacity to metabolize (detoxify) glyphosate. Massive glyphosate use has led to world-wide evolution of glyphosate-resistant (GR) weed species, including the economically damaging grass weed Echinochloa colona. An Australian population of E. colona has evolved resistance to glyphosate with unknown mechanisms that do not involve the glyphosate target enzyme 5-enolpyruvylshikimate-3-P synthase. GR and glyphosate-susceptible (S) lines were isolated from this population and used for resistance gene discovery. RNA sequencing analysis and phenotype/genotype validation experiments revealed that one aldo-keto reductase (AKR) contig had higher expression and higher resultant AKR activity in GR than S plants. Two full-length AKR (EcAKR4-1 and EcAKR4-2) complementary DNA transcripts were cloned with identical sequences between the GR and S plants but were upregulated in the GR plants. Rice (Oryza sativa) calli and seedlings overexpressing EcAKR4-1 and displaying increased AKR activity were resistant to glyphosate. EcAKR4-1 expressed in Escherichia coli can metabolize glyphosate to produce aminomethylphosphonic acid and glyoxylate. Consistent with these results, GR E. colona plants exhibited enhanced capacity for detoxifying glyphosate into aminomethylphosphonic acid and glyoxylate. Structural modeling predicted that glyphosate binds to EcAKR4-1 for oxidation, and metabolomics analysis of EcAKR4-1 transgenic rice seedlings revealed possible redox pathways involved in glyphosate metabolism. Our study provides direct experimental evidence of the evolution of a plant AKR that metabolizes glyphosate and thereby confers glyphosate resistance.

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Aldo-keto Reductase Metabolizes Glyphosate and Confers Glyphosate Resistance in Echinochloa colona.

Continual increases in the human population and growing concerns related to the energy crisis, food security, disease outbreaks, global warming, and other environmental issues require a sustainable solution from nature. One of the promising resources is cyanobacteria, also known as blue-green algae. They require simple ingredients to grow and possess a relatively simple genome. Cyanobacteria are known to produce a wide variety of bioactive compounds. In addition, cyanobacteria’s remarkable growth rate enables its potential use in a wide range of applications in the fields of bioenergy, biotechnology, natural products, medicine, agriculture, and the environment. In this review, we have summarized the potential applications of cyanobacteria in different areas of science and development, especially related to their use in producing biofuels and other valuable co-products. We have also discussed the challenges that hinder such development at an industrial level and ways to overcome such obstacles.

Cyanobacteria: Review of Current Potentials and Applications

The increasing incidence of mass developments of Cyanobacteria in fresh- and brackish water is a matter of growing concern due to the production of toxins that threaten human and livestock health. The toxins that are produced by freshwater Cyanobacteria comprise hepatotoxins (cyclic peptides such as microcystins and nodularin, as well as alkaloids such as cylindrospermopsin) and neurotoxins (alkaloids such as anatoxin-a, anatoxin-a(S) and saxitoxins). The variation in toxicity between and within species of Cyanobacteria has been recognised for a long time. However, the toxic and non-toxic genotypes within a species cannot be discriminated under the microscope, which has been a major obstacle in identifying those factors that influence toxin production both in the laboratory and in the field. During the last decade, major advances were achieved due to the elucidation and functional characterisation of genes, such as the gene cluster encoding the synthesis of the hepatotoxic heptapeptide, microcystin. Genetic techniques, in particular, have been used to explore (i) the genetic basis, biosynthesis pathways, and physiological regulation of toxin (microcystin) production, (ii) gene loss processes resulting in a patchy distribution of the microcystin synthetase gene cluster among genera and species, as well as (iii) the distribution and abundance of the microcystin genes in the environment. In recent years, experience in detecting microcystin genes directly in the field has increased enormously and robust protocols for the extraction of DNA and the subsequent detection of genes by PCR (polymerase chain reaction)based methods are now available. Due to the high sensitivity of PCR, it is possible to detect toxic genotypes long before a toxic cyanobacterial bloom may occur. Consequently, waterbodies that are at risk of toxic bloom formation can be identified early on in the growing season along with environmental factors that can potentially influence the abundance of toxin producing genotypes.

https://www.uibk.ac.at/limno/files/pdf/kurmayer_chritiansenhighres.pdf

The Genetic Basis of Toxin Production in Cyanobacteria

This study presents a phenol and lysozyme free protocol for genomic DNA isolation of cyanobacteria from culture, mats and soil. For an efficient and pure DNA isolation from cyanobacteria having tough cell wall, extra steps of glass beading and Sepharose 4B purification were added. The modified method gave a higher yield of DNA than the phenol: chloroform extraction method. Four parameters selected for purity testing of the isolated DNA were: (i) restriction digestion with Hind III, (ii) randomly amplified polymorphic DNA-PCR of axenic culture of cyanobacteria to assess phylogenetic relatedness, (iii) denaturing gradient gel electrophoretic (DGGE) analysis of cyanobacterial mat and soil to ascertain the applicability of the isolated DNA for community analysis, and (iv) sequencing of partial 16S rDNA of Hapalosiphon intricatus BHULCR1, Anabaena doliolum LCR1, Anabaena oryzae LCR2, Aulosira fertilissima LCR4, and Tolypothrix tenuis LCR7 and BLAST analysis to confirm their cyanobacterial identity. Data generated from above analyses lead us to conclude that the modified method in question is rapid, cost effective, health and time conscious and promising for genetic fingerprinting and community analysis of cyanobacteria from diverse habitats.

A rapid and cost-effective method of genomic DNA isolation from cyanobacterial culture, mat and soil suitable for genomic fingerprinting and community analysis

Present study deals with the identification of a novel aldo/keto reductase, AKR17A1 from Anabaena sp. PCC7120 and adds on as 17th family of AKR superfamily drawn from a wide variety of organisms. AKR17A1 shares many characteristics of a typical AKR such as— (i) conferring tolerance to multiple stresses like heat, UV-B, and cadmium, (ii) excellent activity towards known AKR substrates (isatin and 2-nitrobenzaldehyde), and (iii) obligate dependence on NADPH as a cofactor for enzyme activity. The most novel attribute of AKR17A1, first reported in this study, is its capability to metabolize butachlor, a persistent rice field herbicide that adversely affects agro-ecosystem and non-target organisms. The AKR17A1 catalyzed- degradation of butachlor resulted into formation of 1,2-benzene dicarboxylic acid and 2,6 bis (1,1, dimethylethyl) 4,-methyl phenol as the major products confirmed by GC-MS analysis.

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A Novel Aldo-Keto Reductase (AKR17A1) of Anabaena sp. PCC 7120 Degrades the Rice Field Herbicide Butachlor and Confers Tolerance to Abiotic Stresses in E. coli.

The effect of drought stress on cytosolic and membrane proteome of nitrogen fixing cyanobacterium, Anabaena PCC 7120 was investigated at different time interval using two-dimensional gel electrophoresis and mass spectrometry. > 300 and 140 protein spots in cytosolic and membrane gels respectively detected by proteomic analysis showed reproducible abundance within replications. Of these proteins, 69 and 86 protein spots in cytosolic and membrane proteome respectively displayed differential expression pattern. The survival strategy adopted by Anabaena PCC 7120 under dehydration as presumed by proteomic analysis can be summarized as 1) increased abundance of proteins: Mn catalase, OR, AhpC, SodA, SodB, GST and All1124 to mitigate oxidative damage, 2) minimization of energy expenditure by inhibition of photosynthesis through down-regulation of photosynthetic apparatus, antenna proteins, PSI, PSII and cytochrome b6f and respiration, 3) increased abundance of membrane porins (Omp85, OprB-I, All7614 and Alr4550) and TolC conceivably to support carbohydrate transport for enduring stress and recovery, 4) increase in phosphate transporter (PstS1) to improve phosphate assimilation required to recover cellular damage during rehydration and 5) increased abundance of K+ ATPase during prolonged dehydration and minor abundance of Na+/H+ ATPase during early rehydration to maintain cellular ion homeostasis. Dehydration, however, strongly impaired cytosolic proteins associated with nitrogen fixation, energy metabolism, amino acid and nucleic acid biosynthesis which were resumed after prolonged rewetting. Severely decreased abundance of novel proteins Alr1819, Alr2903, Alr3514, Alr2751 and All3324 serve as the marker for dehydration stress. The above results help in investigating the involvement of the proteins in protection and adaptation mechanisms associated with rejuvenation of the active state of Anabaena sp. PCC 7120.

Dehydration and rehydration - induced temporal changes in cytosolic and membrane proteome of the nitrogen fixing cyanobacterium Anabaena sp. PCC 7120

The present study is an attempt to know the temporal and spatial distribution of geochemical components in the lignite deposit of Kapurdi, Barmer Basin (Rajasthan). Lignite samples have been subjected to proximate, ultimate and elemental analyses, and determination of mineral carbon (MINC%). Besides, various minerals and functional groups have been analyzed through X-ray diffraction (XRD) and Fourier transforms infrared spectroscopy (FTIR). Selected environmentally sensitive and potential hazardous elements like Cu, Cd, Zn, Ni, Cr, Pb, Mn, Al, Fe and Co are determined using atomic absorption spectrophotometer (AAS). To know the association of minerals with organic matter, scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) has also been carried out. The concentration of Co, Ni, Cd, Pb, Na, and K is high when compared with world average and is main concern for the environmental and health hazards. The elements like Fe, Ca, Mg, Zn, and Pb have shown increasing trend from top to bottom of the lignite seam with some fluctuations in the values in few bands whereas others do not follow a definite trend of variation along the seam profile.

Study of minerals and selected environmentally sensitive elements in Kapurdi lignites of Barmer Basin, Rajasthan, western India: implications to environment

The Rasa Bay (North Adriatic, Croatia) has been receiving various pollutants by inflowing streams laden with untreated municipal and coalmine effluents for decades The locality was a regional center of coalmining (Rasa coal), coal combustion, and metal processing industries for more than two centuries As local soil and stream water were found to be contaminated with sulfur and potentially toxic trace elements (PTEs) as a consequence of weathering of Rasa coal and its waste, some clean-up measures are highly required Therefore, the aim of this study was to test the remediating potential of selected microorganisms and synthetic zeolites in the case of soil and coal-mine water, respectively, for the first time By employing bacterial cultures of Ralstonia sp, we examined removal of sulfur and selected PTEs (As, Ba, Co, Cr, Cu, Ni, Pb, Rb, Se, Sr, U, V, and Zn) from soil The removal of sulfur was up to 60%, arsenic up to 80%, while Se, Ba, and V up to 60%, and U up to 20% By applying synthetic zeolites on water from the Rasa coalmine and a local stream, the significant removal values were found for Sr (up to 999%) and Ba (up to 992%) only Removal values were quite irregular (insignificant) in the cases of Fe, Ni, Zn, and Se, which were up to 80%, 50%, 30%, and 20%, respectively Although promising, the results call for further research on this topic

Use of Bacteria and Synthetic Zeolites in Remediation of Soil and Water Polluted with Superhigh-Organic-Sulfur Raša Coal (Raša Bay, North Adriatic, Croatia)

Shweta Rai

Papers

A rapid and cost-effective method of genomic DNA isolation from cyanobacterial culture, mat and soil suitable for genomic fingerprinting and community analysis

A Novel Aldo-Keto Reductase (AKR17A1) of Anabaena sp. PCC 7120 Degrades the Rice Field Herbicide Butachlor and Confers Tolerance to Abiotic Stresses in E. coli.

Dehydration and rehydration - induced temporal changes in cytosolic and membrane proteome of the nitrogen fixing cyanobacterium Anabaena sp. PCC 7120

Study of minerals and selected environmentally sensitive elements in Kapurdi lignites of Barmer Basin, Rajasthan, western India: implications to environment

Use of Bacteria and Synthetic Zeolites in Remediation of Soil and Water Polluted with Superhigh-Organic-Sulfur Raša Coal (Raša Bay, North Adriatic, Croatia)