The zinc finger protein 3 of Arabidopsis thaliana regulates vegetative growth and root hair development.

Introduction: Zinc finger protein 3 (ZFP3) and closely related C2H2 zinc finger proteins have been identified as regulators of abscisic acid signals and photomorphogenic responses during germination. Whether ZFP3 and related ZFP factors regulate plant development is, however, not known. Results: ZFP3 overexpression reduced plant growth, limited cell expansion in leaves, and compromised root hair development. The T-DNA insertion zfp3 mutant and transgenic lines with silenced ZFP1, ZFP3, ZFP4, and ZFP7 genes were similar to wild-type plants or had only minor differences in plant growth and morphology, probably due to functional redundancy. RNAseq transcript profiling identified ZFP3-controlled gene sets, including targets of ABA signaling with reduced transcript abundance. The largest gene set that was downregulated by ZFP3 encoded regulatory and structural proteins in cell wall biogenesis, cell differentiation, and root hair formation. Chromatin immunoprecipitation confirmed ZFP3 binding to several target promoters. Discussion: Our results suggest that ZFP3 and related ZnF proteins can modulate cellular differentiation and plant vegetative development by regulating the expression of genes implicated in cell wall biogenesis.

Double DJ-1 domain containing Arabidopsis DJ-1D is a robust macromolecule deglycase.

Plants, being sessile, are prone to genotoxin-induced macromolecule damage. Among the inevitable damaging agents are reactive carbonyls that induce glycation of DNA, RNA and proteins to result in the build-up of advanced glycated end-products. However, it is unclear how plants repair glycated macromolecules. DJ-1/PARK7 members are a highly conserved family of moonlighting proteins having double domains in higher plants and single domains in other phyla. Here we show that Arabidopsis DJ-1D offers robust tolerance to endogenous and exogenous stresses through its ability to repair glycated DNA, RNA and proteins. DJ-1D also reduced the formation of reactive carbonyls through its efficient methylglyoxalase activity. Strikingly, full-length double domain-containing DJ-1D suppressed the formation of advanced glycated end-products in yeast and plants. DJ-1D also efficiently repaired glycated nucleic acids and nucleotides in vitro and mitochondrial DNA in vivo under stress, indicating the existence of a new DNA repair pathway in plants. We propose that multi-stress responding plant DJ-1 members, often present in multiple copies among plants, probably contributed to the adaptation to a variety of endogenous and exogenous stresses.

Induction of drought tolerance in Pennisetum glaucum by ACC deaminase producing PGPR- Bacillus amyloliquefaciens through Antioxidant defense system.

Rhizobacteria from pearl millet were screened to produce 1-aminocyclopropane-1-carboxylate (ACC) deaminase and to evaluate its role in alleviating drought stress. Amongst 96 isolates, 28 were positive for ACC deaminase production, with MMR04 offering maximum activity of 2196.23 nmol of α-ketobutyrate produced mg-1 of protein h-1. The ACC deaminase producing rhizobacteria with multiple beneficial properties along with root colonization and non-pathogenic were selected [Bacillus amyloliquefaciens (MMR04), Bacillus subtilis (MMR18) and Stenotrophomonas maltophilia (MMR36)] to confirm the presence of ACC deaminase gene. A significant enhancement in seed germination (91.75%) and seedling vigor (1213.73) was noted upon seed treatment with MMR04 and hence further evaluated for its ability to induce drought stress. The seed treatment with MMR04 improved plant growth parameters and total chlorophyll and RWC in plants grown under severe drought stress (G5) conditions compared to control plants. In addition, MMR04 seed treatment enhanced proline, APX and SOD activity while decreased the MDA content up to 2.3 fold compared to untreated plants (G5). Gene expression studies revealed a significant decrease of 3.3 and 1.8 fold in the relative expression of drought-responsive (DREB-1E) and ethylene-responsive factor (ERF-1B) marker genes, respectively and an increase of 2.2 and 2.9 fold in the relative expression of APX1 and SOD1, respectively in MMR04 treated plants grown under G5 conditions over control. The results confirmed that ACC deaminase producing B. amyloliquefaciens MMR04 could defend the pearl millet plants against drought stress through an antioxidative system, thereby warranting its application in drought stress management.

RNA Blot Analysis for the Detection and Quantification of Plant MicroRNAs.

MicroRNAs (miRNAs) are a class of endogenously expressed non-coding, ~21 nt small RNAs involved in the regulation of gene expression in both plants and animals. Most miRNAs act as negative switches of gene expression targeting key genes. In plants, primary miRNAs (pri-miRNAs) transcripts are generated by RNA polymerase II, and they form varying lengths of stable stem-loop structures called pre-miRNAs. An endonuclease, Dicer-like1, processes the pre-miRNAs into miRNA-miRNA* duplexes. One of the strands from miRNA-miRNA* duplex is selected and loaded onto Argonaute 1 protein or its homologs to mediate the cleavage of target mRNAs. Although miRNAs are key signaling molecules, their detection is often carried out by less than optimal PCR-based methods instead of a sensitive northern blot analysis. We describe a simple, reliable, and extremely sensitive northern method that is ideal for the quantification of miRNA levels with very high sensitivity, literally from any plant tissue. Additionally, this method can be used to confirm the size, stability and the abundance of miRNAs and their precursors.

Induction of drought tolerance in tomato upon the application of ACC deaminase producing plant growth promoting rhizobacterium Bacillus subtilis Rhizo SF 48.

A total of ten 1-aminocyclopropane-1-carboxylate (ACC) deaminase producing PGPR isolates were selected and evaluated for the induction of drought stress tolerance in tomato. Among the selected PGPR, maximum seed (laboratory) and plant growth promotion (greenhouse) was observed in tomato seeds bacterized with Bacillus subtilis Rhizo SF 48. The genomic study confirmed the presence of ACC deaminase gene in Rhizo SF 48 and the obtained sequence was deposited to the NCBI database with the Accession No. MK652706. The tomato plants grown upon treatment with Rhizo SF 48 significantly enhanced plant growth even after exposing to different levels of drought stress as compared to stress induced control plants. About 7.5% and 38% increase in RWC were observed in Rhizo SF 48 treated tomato plants grown under well-watered and stress conditions (S4) compared to their control plants, respectively. An increase of 0.76, 0.23 and 0.78 fold in proline, SOD and APX activity and a decrease of 0.3 fold in MDA and H2O2 contents were observed in Rhizo SF 48 treated plants compared to control plants grown under S4 conditions. The histo-chemical studies showed lower accumulations of H2O2 and superoxide anion in the leaves of Rhizo SF 48 treated plants under drought stress, which was in confirmation with the quantification results of H2O2 and SOD. The qRT-PCR studies on drought (Le25) and ethylene responsive factor (SlERF84) marker genes showed that a significant decrease of 0.75 and 0.81 folds, respectively in Le25 and SlERF84 accumulation was observed in Rhizo SF 48 treated plants compared to untreated plants grown under S4 conditions. From the results, it can be attributed that ACC deaminase producing Rhizo SF 48 was able to protect tomato plants against oxidative damage caused due to drought stress and enhanced plant growth promotion. It can be concluded that ACC deaminase producing Rhizo SF 48 can serve as a useful bio-inoculant for sustainable tomato production in arid and semi-arid regions with water deficit.

Major Domestication-Related Phenotypes in Indica Rice Are Due to Loss of miRNA-Mediated Laccase Silencing.

Domestication of rice (Oryza sativa) included conversion of perennial wild species with few seeds to short plants that produced abundant seeds. Most domestication-associated changes were due to variations in transcription factors and other key proteins such as enzymes. Here, we show that multiple yield-related traits associated with indica rice domestication are linked to micro (mi) RNA-mediated regulation. Analysis of small (s) RNA data sets from cultivated indica rice lines, a few landraces, and two wild relatives of rice revealed the presence of abundant 22-nucleotide (nt) reads in wild relatives that mapped to miR397 precursors. miR397 was expressed at very high levels in wild relatives and at negligible levels in high-yielding cultivated lines. In its genera-specific form of 22-nt, miR397 targeted mRNAs encoding laccases that decayed and induced robust secondary cascade silencing in wild species that required RNA-dependent RNA polymerase 6. In wild species of rice, reduced expression of laccases resulted in low lignification. As expected, overexpression of miR397 induced de-domestication phenotypes. At least 26 uncharacterized QTLs previously implicated in rice yield overlapped with laccases and miR397 genes. These results suggest that miRNAs contribute to rice domestication-associated phenotypes.

Novel Organotin(IV)-Schiff Base Complexes: Synthesis, Characterization, Antimicrobial Activity, and DNA Interaction Studies.

Four organotin(IV) complexes with 2-(2-hydroxybenzylideneamino)isoindoline-1,3-dione (L(1)), and 4-(4-hydroxy-3-methoxybenzylideneamino-N-(pyrimidin-2-yl)benzenesulfonamide (L(2)) were synthesized and well characterized by analytical and spectral studies. The synthesized compounds were tested for antimicrobial activity by disc diffusion method. The DNA binding of the complexes 1 and 3 with CT-DNA has been performed with absorption spectroscopy, which showed that both the complexes are avid binders of CT-DNA. Also the nuclease activity of complexes 1 and 3 with plasmid DNA (pUC19) was studied using agarose gel electrophoresis. The complex 1 can act as effective DNA cleaving agent when compared to complex 3 resulting in the nicked form of DNA under physiological conditions. The gel was run both in the absence and presence of the oxidizing agent.