Transcriptome Analysis for Abiotic Stresses in Rice (<em>Oryza sativa</em> L.)

Rice, a model monocot system, belongs to the family Poaceae and genus Oryza. Rice is the second largest produced cereal and staple food crop fulfilling the demand of half the world’s population. Though rice demand is growing exponentially, its production is severely affected by variable environmental changes. The various abiotic factors drastically reduce the rice plant growth and yield by affecting its different growth stages. To fulfill the growing demand of rice, it is imperative to understand its molecular responses during stresses and to develop new varieties to overcome the stresses. Earlier, the microarray experiments have been used for the identification of coexpressive gene networks during various conditions in crop plants. Though the microarray experiments provided very useful information, the unviability of genome-wide information did not provide complete information about the regulatory gene networks involved in the stress response. The advancement of molecular techniques provided breakthrough to understanding the complex regulatory gene networks and their signaling pathways during stresses. The high-throughput RNA sequencing data have opened the floodgate of transcriptome data in rice. Here we have summarized some of the transcriptome data for abiotic molecular responses in rice, which further help to understand their complex regulatory mechanism

Isolation, cloning and characterization of phlB gene from an Indian strain of Gram negative soil bacteria Pseudomonas fluorescens

412-419Phloroglucinol (PHL) or 2,4-diacetylphloroglucinol (DAPG) is a polyketide compound that exhibits broad spectrum anti-bacterial and antifungal properties against soil-borne plant pathogens. It is produced by gram negative bacterium Pseudomonas (fluorescent Pseudomonad) which renders them a potential biological control agent. The process of PHL production is accomplished by a well coordinated activity of four (phlABCD) genes arranged in an operon. All four genes are essential and indispensable for biosynthesis of DAPG from precursor malonyl CoA. Here, we report cloning and analysis of phlB gene from an Indian strain of Pseudomonas fluorescens. Further, we predicted the structure of PHLB protein and analyzed its intrinsic details. Stereo-chemical properties were analyzed by Ramachandran plot and secondary structure was retrieved by PDB sum. Our study provides insight into the structure of PHLB protein and revealed that though Pseudomonas PHLB lacks conserved motifs and domains it is indispensable for monoacetyl phloroglucinol (MAPG) synthesis

Exceptional molecular and coreceptor-requirement properties of molecular clones isolated from an Human Immunodeficiency Virus Type-1 subtype C infection

<p>Abstract</p> <p>Background</p> <p>The pathogenic significance of coreceptor switch in the viral infection of HIV-1 is not completely understood. This situation is more complex in subtype C infection where coreceptor switch is either absent or extremely rare. To gain insights into the mechanisms that underlie coreceptor requirement of subtype C, we screened several primary viral isolates and identified a clinical sample that demonstrated a potential to grow on standard T-cell lines with no detectable CCR5 expression. The subject was diagnosed with HIV-1 associated dementia in the absence of opportunistic infections of the brain. To isolate molecular clones from this virus, we devised a novel strategy based on anchor primers that target a sequence in the reverse transcriptase, highly conserved among diverse subtypes of HIV-1.</p> <p>Results</p> <p>Using this strategy, we isolated 8 full-length molecular clones from the donor. Two of the eight molecular clones, 03In94_D17 and 03In94_D24, (D17 and D24) generated replication-competent viruses. Phylogenetic analysis of the full-length viral sequences revealed that both clones were non-recombinant subtype C viruses. They contain intact open reading frames in all the viral proteins. Both the viral clones are endowed with several unique molecular and biological properties. The viral promoter of the clones is characterized by the presence of four NF-kB binding elements, a feature rarely seen in the subtype C HIV-1 LTR. Interestingly, we identified the coexistence of two different forms of Rev, a truncated form common to subtype C and a full-length form less common for this subtype, in both proviral and plasma virus compartments. An exceptional property of the viruses, atypical of subtype C, is their ability to use a wide range of coreceptors including CCR5, CXCR4, and several others tested. Sequence analysis of Env of D17 and D24 clones identified differences within the variable loops providing important clues for the expanded coreceptor use. The V1, V2 and V4 loops in both of the molecular clones are longer due to the insertion of several amino acid residues that generated potential N-linked glycosylation sites.</p> <p>Conclusion</p> <p>The exceptional biological and molecular properties of these clones make them invaluable tools to understand the unique pathogenic characteristics of subtype C.</p

Drought Induced Signaling in Rice: Delineating Canonical and Non-canonical Pathways

Drought induced stress is often a bottleneck of agricultural crop production. Invariably, field crops across all agro-ecological regions succumb to it with an yield penalty. Drought massively affects the growth and harvestable yield in crops and has become an imminent problem necessitating breeding of tolerant crops. It induces myriad changes of biochemical, molecular, and physiological nature that manifest into aberrant plant morphology. The response to drought in plants incites a signaling cascade that involves perception and translation of drought signal leading to concomitant modulation of gene expression and de novo osmolyte synthesis. The intricate patterns of expression of these genes vary from early induction to late responsive genes. While one class of genes codes for products imparting osmotolerance and protection to plants, the second class predominantly modulates target gene expression by an intricate signal transduction mechanism. This review summarizes both canonical and non-canonical cascades of drought stress response in plants, delineating the mechanism in rice (Oryza sativa) and emphasizes hydropenia induced lipid signaling

Cloning and characterization of phloroglucinol biosynthetic gene phlC from an Indian strain of Pseudomonas fluorescens

Phloroglucinol or 2,4-diacetyl phloroglucinol (DAPG) is a polyketide compound produced by gram negative soil bacteria Pseudomonas. It shows broad spectrum antibacterial and antifungal properties against soil-borne plant pathogens. In Pseudomonas spp., genes for biosynthesis of 2,4-DAPG are localized in phlABCD operon. All the four genes in phlABCD operon are indispensable and DAPG synthesis is attenuated even in the absence of one of the genes. In the present study, we identified and cloned phlC gene from an Indian strain of Pseudomonas and analyzed its sequence. The structural details ofthe PHLC protein was generated by three-dimensional homology modelling. Additionally, stereo-chemical properties of PHLC were analyzed by Ramachandran plot analysis and the generated model was validated by PDBsum. Our results demonstrate that the cloned PHLC protein contains structural features typical of a condensing enzyme involved inpolyketide synthesis

Cloning and structural elucidation of a brassinosteroids biosynthetic gene (Atdwarf4) and genetic transformation of Indian mustard (Brassica juncea L.)

320-330Phytohormones play critical roles in plant growth and development. Brassinosteroids (BRs) are essential group of phytohormones required for optimum growth of plants and their deficiency causes distinctive dwarf phenotypes in plants. Homeostasis of BRs in plants is maintained by DWARF4 enzyme that mediates multiple 22α-hydroxylation steps in brassinosteroid biosynthesis. Arabidopsis plants over-expressing DWARF4 show increase in inflorescence, number of branches and siliques; thereby increased number of seeds/plant. This suggests that engineering DWARF4 biosynthesis in Brassica plant can be strategized to enhance yield in mustard. In the present study (i) we cloned dwarf4 gene from Arabidopsis using gene specific PCR strategy, (ii) elucidated the three-dimensional structure of DWARF4 protein at molecular level where it revealed presence of four beta sheets and 20 alpha-helices, and (iii) transformed mustard cultivar Pusa Jaikisan with an objective to develop transgenic mustard with enhanced number of siliques. We obtained several putative transgenics with an average transformation efficiency of 3.3%. Molecular characterization with nptII specific primers confirmed presence of transgene in six putative transgenic plants

Immune Activations and Viral Tissue Compartmentalization During Progressive HIV-1 Infection of Humanized Mice

Human immunodeficiency virus type one (HIV-1) tissue compartments are established soon after viral infection. However, the timing in which virus gains a permanent foothold in tissue and the cellular factors that control early viral-immune events are incompletely understood. These are critical events in studies of HIV-1 pathogenesis and in the development of viral reservoirs after antiretroviral therapy. Moreover, factors affecting the permanence of viral-tissue interactions underlie barriers designed to eliminate HIV-1 infection. To this end we investigated the temporal and spatial viral and host factors during HIV-1 seeding of tissue compartments. Two humanized NOD.Cg-Prkdcscid IL2rgtm1Wjl/SzJ mouse models were employed. In the first, immune deficient mice were reconstituted with human CD34+ cord blood hematopoietic stem cells (HSC) (hu-HSC) and in the second mice were transplanted with adult mature human peripheral lymphocytes (hu-PBL). Both, in measure, reflect relationships between immune activation and viral infection as seen in an infected human host. Following humanization both mice models were infected with HIV-1ADA at 104 50% tissue culture infective doses. Viral nucleic acids and protein and immune cell profiles were assayed in brain, lung, spleen, liver, kidney, lymph nodes, bone marrow, and gut from 3 to 42 days. Peripheral CD4+ T cell loss began at 3 days together with detection of HIV-1 RNA in both mouse models after initiation of HIV-1 infection. HIV-1 was observed in all tested tissues at days 3 and 14 in hu- PBL and HSC mice, respectively. Immune impairment was most prominent in hu-PBL mice. T cell maturation and inflammation factors were linked directly to viral tissue seeding in both mouse models. We conclude that early viral tissue compartmentalization provides a roadmap for investigations into HIV-1 elimination

Multipolymer microsphere delivery of SARS-CoV-2 antigens

Effective antigen delivery facilitates antiviral vaccine success defined by effective immune protective responses against viral exposures. To improve severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) antigen delivery, a controlled biodegradable, stable, biocompatible, and nontoxic polymeric microsphere system was developed for chemically inactivated viral proteins. SARS-CoV-2 proteins encapsulated in polymeric microspheres induced robust antiviral immunity. The viral antigen-loaded microsphere system can preclude the need for repeat administrations, highlighting its potential as an effective vaccine. Statement of significance Successful SARS-CoV-2 vaccines were developed and quickly approved by the US Food and Drug Administration (FDA). However, each of the vaccines requires boosting as new variants arise. We posit that injectable biodegradable polymers represent a means for the sustained release of emerging viral antigens. The approach offers a means to reduce immunization frequency by predicting viral genomic variability. This strategy could lead to longer-lasting antiviral protective immunity. The current proof-of-concept multipolymer study for SARS-CoV-2 achieve these metrics. [PDF also includes a graphical abstract that can not be displayed here.