Oryza Sativa Cv Research Articles

Transcriptional and metabolic profiling of sulfur starvation response in two monocots

BackgroundSulfur (S) is a mineral nutrient essential for plant growth and development, which is incorporated into diverse molecules fundamental for primary and secondary metabolism, plant defense, signaling, and maintaining cellular homeostasis. Although, S starvation response is well documented in the dicot model Arabidopsis thaliana, it is not clear if the same transcriptional networks control the response also in the monocots.ResultsWe performed series of physiological, expression, and metabolite analyses in two model monocot species, one representing the C3 plants, Oryza sativa cv. kitaake, and second representing the C4 plants, Setaria viridis. Our comprehensive transcriptomic analysis revealed twice as many differentially expressed genes (DEGs) in S. viridis than in O. sativa under S-deficiency, consistent with a greater loss of sulfur and S-containing metabolites under these conditions. Surprisingly, most of the DEGs and enriched gene ontology terms were species-specific, with an intersect of only 58 common DEGs. The transcriptional networks were different in roots and shoots of both species, in particular no genes were down-regulated by S-deficiency in the roots of both species.ConclusionsOur analysis shows that S-deficiency seems to have different physiological consequences in the two monocot species and their nutrient homeostasis might be under distinct control mechanisms.

Cereals can trap endophytic bacteria with potential beneficial traits when grown ex-situ in harsh soils.

Microbial communities associated with plants growing in harsh conditions, including salinity and water deficiency, have developed adaptive features which permit them to grow and survive under extreme environmental conditions. In the present study, an ex-situ plant trapping method has been applied to collect the culturable microbial diversity associated with the soil from harsh and remote areas. Oryza sativa cv. Baldo and Triticum durum Primadur plants were used as recruiters, while the soil surrounding the roots of Oryza glaberrima plants from remote regions of Mali (West Africa) was used as substrate for their growth. The endophytic communities recruited by the two plant species belonged to Proteobacteria and Firmicutes, and the dominant genera were Bacillus, Kosakonia, and Enterobacter. These endophytes were characterized by analyzing some of the most common plant growth promoting traits. Halotolerant, inorganic phosphate-solubilizing and N-fixing strains were found, and some of them simultaneously showing these three traits. We verified that 'Baldo' recruited mostly halotolerant and P-solubilizers endophytes, while the endophytes selected by 'Primadur' were mainly N-fixers. The applied ex-situ plant trapping method allowed to isolate endophytes with potential beneficial traits that could be applied for the improvement of rice and wheat growth under adverse environmental conditions.

Assessment of Nutrient Minerals and Potentially Toxic Elements in a Bangladesh-Origin Rice Cultivar (Oryza sativa cv. BR11) and Associated Health

Rice contributes significantly to both food security and livelihoods in Bangladesh. Despite challenges such as an immense population growth rate and decreased arable land since gaining independence, Bangladesh has attained self-sufficiency in rice production by adopting modern rice varieties (MRVs). However, not all introduced MRVs have gained widespread acceptance among farmers due to concerns about productivity and profitability. Notably, BR11, an MRV, stands out as a prominently adopted rice cultivar during the Aman rice-growing season. This study aims to assess the levels of essential nutrient minerals (Na, K, Ca and Mg) and potentially toxic elements (PTEs) (Cd, Cu, Pb, Ni, Co, Fe, Mn, Zn, Cr) in the BR11 rice cultivar and also to evaluate associated health risks. The concentrations of Na, K, Ca and Mg (mg 100 g–1) in the sampled BR11 rice cultivar were found to be 4.4 ± 2.4, 108.5 ± 24.9, 3.4 ± 0.4 and 4.5 ± 1.1, respectively. Among the PTEs, Cd, Pb, Ni, Co, or Cr contents were below the detection limit, while Fe, Mn, Zn and Cu contents (mg kg–1) were 12.0 ± 4.0, 9.2 ± 1.5, 18.7 ± 2.7 and 2.9 ± 1.0. Comparisons of PTE contents in BR11 rice cultivars exposed to industrial activities with those cultivated in non-industrial zones did not reveal any statistically significant variations (p < 0.05). The estimated daily intake of nutrient minerals through the consumption of BR11 rice was significantly below the daily recommended dietary reference intakes. Likewise, the accumulation of PTEs in the BR11 rice cultivar was well below the maximum allowable concentrations recommended by WHO or FAO. Health risk assessments, involving the computation of non-carcinogenic and carcinogenic risks, did not indicate any concerning risk scenarios associated with the consumption of the BR11 rice cultivar. The data suggests that BR11 is a safe MRV for daily consumption, affirming its suitability for inclusion in the daily diet.

Effect of Priming on Seed Germination Characteristics of Spring Rice (Oryza sativa cv. Hardinath Hybrid-1) in Chandrapur, Rautahat, NEPAL

To determine the effects of different priming on Germination and Seedling Characteristics of Spring rice (Oryza Sativa L.), an experiment was carried out at Chandrapur municipality-4, Rautahat during March of 2022 with Seven different treatments viz. T1 Potash (3% Solution), T2 Urea (3% Solution), T3 DAP (3% Solution), T4 Halo (1Mole Nacl/Litre), T5 Zinc (3% Solution), T6 Hydro and T7 Control. The experiment was laid out in simple CRD design with three replications. Hardinath Hybrid-1 Variety was used for the experiment. Observations on Germination %, Germination Energy (%), Speed of Germination (%), Vigor Index, Root and Shoot Length, Fresh Weight of Seedlings and Dry Weight of Seedlings were considered. Data was entered and tabulated using MS-Excel, while Analysis of variation and mean separation was done using R-Studio. Biometrical Observation like germination Energy was found highest under the treatment DAP (82.67). The germination percentage (93.00) and vigor index (161.4) was found highest in DAP (3%) Priming and speed of germination (86.84) was found highest in Potash Priming. Similarly, root length (6.14cm) and shoot length (6.23cm) was found to be highest in DAP treatment followed by Halo treatment with root length (5.30cm) and shoot length (5.44 cm). At 10th and 15th Day, there is significant effect on root length while there are no significant differences/effect on Shoot length, fresh weight and Dry Weight. Among these priming treatments, the highest root length was found in priming of DAP; 7.84 cm and 8.07 cm.

Rice straw-derived smoke water promotes rice root growth under phosphorus deficiency by modulating oxidative stress and photosynthetic gene expression

Plant-derived smoke has been shown to promote plant growth and seed germination, but its roles and mechanisms in response to nutrient deficiency stress remain unclear. Plants respond to phosphorus (P) deficiency by undergoing morphological, physiological, and transcriptional changes in order to improve nutrient uptake efficiency. Here, we showed that rice straw-derived smoke water could promote root growth in rice (Oryza sativa cv. Nipponbare) grown under P-sufficient and P-deficient conditions. Transcriptome analysis of the root tissues identified 1309 genes up-regulated and 1311 genes down-regulated by smoke water under P-deficient conditions. The GO terms ‘glutathione transferase activity’ and ‘photosynthesis—light reaction’ were found to be significantly enriched among the genes that were up- and down-regulated by smoke water, respectively. Biochemical analysis showed that smoke water reduced P-deficient-induced accumulation of H2O2 and malondialdehyde (MDA), a lipid peroxidation marker, reduced sucrose contents, but increased Fe accumulation. Furthermore, smoke water suppressed the expression of strigolactone biosynthesis genes, which were strongly induced by P deficiency as an adaptive strategy to improve root P uptake. These results revealed a potential mechanism by which smoke water promotes root growth and interacts with P deficiency-induced transcriptional regulation to mitigate P deficiency stress in rice.

Spatially resolved gene regulatory networks in Asian rice (Oryza sativa cv. Nipponbare) leaves.

Transcriptome profiles in plants are heterogenous at every level of morphological organization. Even within organs, cells of the same type can have different patterns of gene expression depending on where they are positioned within tissues. This heterogeneity is associated with non-uniform distribution of biological processes within organs. The regulatory mechanisms that establish and sustain the spatial heterogeneity are unknown. Here, we identify regulatory modules that support functional specialization of different parts of Oryza sativa cv. Nipponbare leaves by leveraging transcriptome data, transcription factor binding motifs and global gene regulatory network prediction algorithms. We generated a global gene regulatory network in which we identified six regulatory modules that were active in different parts of the leaf. The regulatory modules were enriched for genes involved in spatially relevant biological processes, such as cell wall deposition, environmental sensing and photosynthesis. Strikingly, more than 86.9% of genes in the network were regulated by members of only five transcription factor families. We also generated targeted regulatory networks for the large MYB and bZIP/bHLH families to identify interactions that were masked in the global prediction. This analysis will provide a baseline for future single cell and array-based spatial transcriptome studies and for studying responses to environmental stress and demonstrates the extent to which seven coarse spatial transcriptome analysis can provide insight into the regulatory mechanisms supporting functional specialization within leaves.

Determination of genome size variation among varieties of Ilex cornuta (Aquifoliaceae) by fow cytometry

Ilex cornuta Lindl. & Paxton is a commercially important horticultural species worldwide, and extensive cultivation and hybridization have produced many varieties. Despite the considerable breeding, selection, widespread cultivation and domestication, which may have a significant role in the composition of genomes, there are no other previous reports of intraspecific genome size variation in the different cultivars or hybrids of this species. In the present work, genome size of 12 varieties of I. cornuta was assessed and analyzed through high-resolution flow cytometry (FCM). Nuclear DNA was analyzed using nuclei isolated from young leaves, which used propidium iodide (PI) staining, with rice (Oryza sativa cv. Nipponbare) as internal reference. As a result, statistically significant differences in genome size were detected among all diploid I. cornuta varieties considered. The estimated genome size (2C value) of I. cornuta varieties ranged from 1.47 to 1.80 pg, with 1.22-fold variation and an average size of 1.65 pg. The domestication and interspecific hybridization induced variation of genome size in I. cornuta, and the genome size of hybrids exhibited a wider range of variation compared with that of cultivars. In summary, flow cytometry is a useful tool to analyze the genome size of I. cornuta. The first report of the genome sizes of varieties of this species would provide useful data for further research on I. cornuta, and enrich the C value database of Ilex L. What’s more, our findings could be the foundation in the future of I. cornuta genome sequencing and breeding programs.

Insight into the physiological and biochemical mechanisms of biostimulating effect of Ascophyllum nodosum and Moringa oleifera extracts to minimize cadmium-induced oxidative stress in rice.

Cadmium (Cd) is a serious threat for environmental sustainability as it can be taken up quickly by plants and transported to the food chain of living organisms. It alters plants' metabolic and physiological activities and causes yield loss, thereby, enhancing plant tolerance to Cd stress is of utmost essential. Therefore, an experiment was executed to investigate the potential role of Ascophyllum nodosum extract (ANE) and moringa (Moringa oleifera) leaf extract (MLE) to confer Cd tolerance in rice (Oryza sativa cv. BRRI dhan89). Thirty-five-day-old seedling was subjected to Cd stress (50mgkg-1 CdCl2) alone and in a combination of ANE (0.25%) or MLE (0.5%) in a semi-controlled net house. Exposure to Cd resulted in accelerated production of reactive oxygen species, enhanced lipid peroxidation, and disrupted antioxidant defense and glyoxalase system, thus retarded plant growth, biomass production, and yield attributes of rice. On the contrary, the supplementation of ANE or MLE enhanced the contents of ascorbate and glutathione, and the activities of antioxidant enzymes such as ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase, glutathione reductase, glutathione peroxidase, and catalase. Moreover, supplementation of ANE and MLE enhanced the activities of glyoxalase I and glyoxalase II which prevented the overgeneration of methylglyoxal in Cd stressed rice plants. Thus, because of ANE and MLE addition Cd-induced rice plants showed a noticeable declination in membrane lipid peroxidation, hydrogen peroxide generation, and electrolyte leakage, whereas improved water balance. Furthermore, the growth and yield attributes of Cd-affected rice plants were improved with the supplementation of ANE and MLE. All the studied parameters indicates the potential role of ANE and MLE in mitigating Cd stress in rice plants through improving the physiological attributes, modulating antioxidant defense and glyoxalase system.

Unique and Shared Proteome Responses of Rice Plants (Oryza sativa) to Individual Abiotic Stresses.

Food safety of staple crops such as rice is of global concern and is at the top of the policy agenda worldwide. Abiotic stresses are one of the main limitations to optimizing yields for sustainability, food security and food safety. We analyzed proteome changes in Oryza sativa cv. Nipponbare in response to five adverse abiotic treatments, including three levels of drought (mild, moderate, and severe), soil salinization, and non-optimal temperatures. All treatments had modest, negative effects on plant growth, enabling us to identify proteins that were common to all stresses, or unique to one. More than 75% of the total of differentially abundant proteins in response to abiotic stresses were specific to individual stresses, while fewer than 5% of stress-induced proteins were shared across all abiotic constraints. Stress-specific and non-specific stress-responsive proteins identified were categorized in terms of core biological processes, molecular functions, and cellular localization.

Studies on genetic variability, heritability and genetic advance of EMS induced mutant population in paddy (Oryza sativa) cv. BPT 5204

Genetic variability in plant population is crucial for crop improvement. Induced mutation is one of the potential tools for generating more variability which has been successfully exploited for improving several traits in many crop plants including rice. This research was conducted during kharif 2021 at Centurion University of Technology and Management, Odisha. This study was aimed at determining the selection criteria for M2 generation of rice variety BPT 5204 through genetic variability, heritability and genetic advance analysis. Analysis of variance indicated that the mean sum of squares due to genotypes was significant for all the characters (P greater than 0.05 and P greater than 0.01) which revealed that there was considerable genetic variability amongst the material under study. The phenotypic coefficient of variation was significantly higher in magnitude than as usual of genotypic coefficient of variation for all the traits under each analysis. High heritability (above 60%) was observed for plant height (75.93%), number of tillers/hill (64.20%), productive tillers/hill (81.54%), sterile grains/plant (92.28%), panicle length (69.49%), flag leaf length (63.29%), flag leaf width (89.77%) and grain yield/hill (68.68%). Genetic advance as per cent of mean was high for traits number of tillers per hill, productive tillers/hill, sterile grains/plant, panicle length, flag leaf length and flag leaf width.

CRISPR/Cas9-Mediated Genome Editing of Soluble Starch Synthesis Enzyme in Rice for Low Glycemic Index

In the present study, the selected rice (Oryza sativa cv. indica) was genetically modified to increase the amylose content in the seeds. The ‘CRISPR-Cas9’ genome-editing tool was used to knock out three isoforms of soluble starch synthase (SSS) viz. SSSII-1, SSSII-2 and SSSII-3. A genetic transformation vector designed with appropriate gRNAs, Cas9, and antibiotic resistance was used to create SSS knockout mutants to enrich the content of amylose. Putative rice mutants were developed with high amylose content in the seeds of up to 63% as compared to 23% in the wild types (control). Rice with a low Glycemic Index (GI) value and high amylose content rice is preferred to avoid a sudden rise in glucose in the bloodstream. The frequencies of bi-allelic or homozygous transgenic lines of SSSII-1, SSSII-2, and SSSII-3 in the first generation were tested via the Mendelian fashion of segregated bi-allelic lines in the T1 generation of the putative rice mutants. The T1 generation segregation showed a frame-shift mutation. A molecular characterization of the putative mutants successfully demonstrated the development of a Cas9-free rice mutant with a higher amount of amylose in the rice.

Whole-Genome Sequencing of Rice Mutant Library Members Induced by N-Methyl-N-Nitrosourea Mutagenesis of Fertilized Egg Cells

Although targeted genome editing technology has become a powerful reverse genetic approach for accelerating functional genomics, conventional mutant libraries induced by chemical mutagens remain valuable for plant studies. Plants containing chemically induced mutations are simple yet effective genetic tools that can be grown without regard for biosafety issues. Whole-genome sequencing of mutant individuals reduces the effort required for mutant screening, thereby increasing their utility. In this study, we sequenced members of a mutant library of Oryza sativa cv. Nipponbare derived from treating single fertilized egg cells with N-methyl-N-nitrosourea (MNU). By whole-genome sequencing 266 M1 plants in this mutant library, we identified a total of 0.66 million induced point mutations. This result represented one mutation in every 146-kb of genome sequence in the 373 Mb assembled rice genome. These point mutations were uniformly distributed throughout the rice genome, and over 70,000 point mutations were located within coding sequences. Although this mutant library was a small population, nonsynonymous mutations were found in nearly 61% of all annotated rice genes, and 8.6% (3248 genes) had point mutations with large effects on gene function, such as gaining a stop codon or losing a start codon. WGS showed MNU-mutagenesis using rice fertilized egg cells induces mutations efficiently and is suitable for constructing mutant libraries for an in silico mutant screening system. Expanding this mutant library and its database will provide a useful in silico screening tool that facilitates functional genomics studies with a special emphasis on rice.

Effects of Storage Temperature on Indica-Japonica Hybrid Rice Metabolites, Analyzed Using Liquid Chromatography and Mass Spectrometry.

The Yongyou series of indica-japonica hybrid rice has excellent production potential and storage performance. However, little is known about the underlying mechanism of its storage resistance. In this study, Yongyou 1540 rice (Oryza sativa cv. yongyou 1540) was stored at different temperatures, and the storability was validated though measuring nutritional components and apparent change. In addition, a broad-targeted metabolomic approach coupled with liquid chromatography-mass spectrometry was applied to analyze the metabolite changes. The study found that under high temperature storage conditions (35 °C), Yongyou 1540 was not significantly worse in terms of fatty acid value, whiteness value, and changes in electron microscope profile. A total of 19 key differential metabolites were screened, and lipid metabolites related to palmitoleic acid were found to affect the aging of rice. At the same time, two substances, guanosine 3′,5′-cyclophosphate and pipecolic acid, were beneficial to enhance the resistance of rice under harsh storage conditions, thereby delaying the deterioration of its quality and maintaining its quality. Significant regulation of galactose metabolism, alanine, aspartate and glutamate metabolism, butyrate metabolism, and arginine and proline metabolism pathways were probably responsible for the good storage capacity of Yongyou 1540.

The first long-read nuclear genome assembly of Oryza australiensis, a wild rice from northern Australia

Oryza australiensis is a wild rice native to monsoonal northern Australia. The International Oryza Map Alignment Project emphasises its significance as the sole representative of the EE genome clade. Assembly of the O. australiensis genome has previously been challenging due to its high Long Terminal Repeat (LTR) retrotransposon (RT) content. Oxford Nanopore long reads were combined with Illumina short reads to generate a high-quality ~ 858 Mbp genome assembly within 850 contigs with 46× long read coverage. Reference-guided scaffolding increased genome contiguity, placing 88.2% of contigs into 12 pseudomolecules. After alignment to the Oryza sativa cv. Nipponbare genome, we observed several structural variations. PacBio Iso-Seq data were generated for five distinct tissues to improve the functional annotation of 34,587 protein-coding genes and 42,329 transcripts. We also report SNV numbers for three additional O. australiensis genotypes based on Illumina re-sequencing. Although genetic similarity reflected geographical separation, the density of SNVs also correlated with our previous report on variations in salinity tolerance. This genome re-confirms the genetic remoteness of the O. australiensis lineage within the O. officinalis genome complex. Assembly of a high-quality genome for O. australiensis provides an important resource for the discovery of critical genes involved in development and stress tolerance.

Facilitation of auxin biosynthesis and metabolism by salt stress in rice plants

To investigate the effects of salt stress on the auxin in rice plants, we analyzed indole-3-acetic acid (IAA) and its metabolites in hydroponically grown rice plant seedlings (Oryza sativa cv. Nipponbare) and in the culture medium using LC-ESI/MS/MS. We found that the IAA level in the culture medium was increased 39-fold due to salt stress treatment, while those in the shoots and roots were increased by less than 2-fold. The culture medium also contained high amounts of oxidized metabolites such as OxIAA, DiOxIAA, DiOxIAA-Glu, and DiOxIAA-Asp. The total amount of IAA and its metabolites in the shoots, roots, and culture medium was significantly increased with salt treatment. A salt stress-responsive increase in the IAA metabolites was observed in 9 tested cultivars of rice. The results indicate that salt stress facilitates the metabolic turnover of IAA in rice plants and may open new insight into the role of auxin.

Characterization and biological activities of melanin pigment from root endophytic fungus, Phoma sp. RDSE17.

Melanins are high molecular weight hydrophobic pigments which have gained popularity for their role in virulence against different pathogens. In the present study, we isolated and characterized the melanin pigment produced by a dark septate endophyte fungus Phoma sp. RDSE17, which was associated with the roots of an indigenous Oryza sativa cv. 'Chakhao amubi' in Manipur, Northeast India. The biological properties of purified melanin from the fungus were evaluated for their antioxidant, antimicrobial and anticancerous activities. The pigment was extracted from Phoma sp. by alkaline-acid hydrolysis method and confirmed as melanin through physico-chemical tests and spectral (UV, FTIR, and EPR) analysis. The analyses of the elemental composition indicated that the pigment possessed a low percentage of nitrogen (N) contents, and therefore, would not fall under DOPA class of melanin. Exposure of the fungus to melanin pathway inhibitors revealed a positive melanin inhibition by tricyclazole, but not by kojic acid. Thus, the melanin from Phoma sp. may be a member of the DHN family. Moreover, the purified melanin showed high DPPH (1, 1-Diphenyl-2-picrylhydrazyl) free radical-scavenging activity with an EC50 of 69µg/mL and inhibited human lung cancer cell (A549 cells) proliferation at 80µg/mL. The present study demonstrates that melanin from Phoma sp. RDSE17 could be employed as a potential biological (antioxidant) and antimicrobial agent for inhibiting the growth of humans and phytopathogens.

Antioxidation, Anti-Inflammation, and Regulation of SRD5A Gene Expression of Oryza sativa cv. Bue Bang 3 CMU Husk and Bran Extracts as Androgenetic Alopecia Molecular Treatment Substances.

Androgenetic alopecia (AGA), a hair loss disorder, is a genetic predisposition to sensitive androgens, inflammation, and oxidative stress. Unfortunately, current treatments with synthetic medicines contain a restricted mechanism along with side effects, whereas the bioactive constituents of plant extracts are multifunctional, with fewer side effects. The massive amounts of rice husk and bran are agricultural wastes that may cause pollution and environmental problems. Owing to these rationales, the local rice variety, Bue Bang 3 CMU (BB3CMU), which is grown in northern Thailand, was evaluated for the valuable utilization of rice by-products, husk (BB3CMU-H) and bran (BB3CMU-RB) extracts, for AGA treatment regarding antioxidant, anti-inflammatory, anti-androgenic activities, and the characterization of bioactive compounds. Our study verified that BB3CMU-H had the highest level of polyphenols, contributing to its greater antioxidant activity. Conversely, BB3CMU-RB was the predominant source of tocopherols, resulting in better anti-androgenic activities regarding the downregulation of steroid 5α-reductase genes (SRD5A). Notably, anti-inflammation via the attenuation of nitric oxide productions was observed in BB3CMU-H (0.06 ± 0.13 μM) and BB3CMU-RB (0.13 ± 0.01 μM), which were significantly comparable to diclofenac sodium salt (0.13 ± 0.19 μM). Therefore, the combination of BB3CMU-H and BB3CMU-RB could be utilized in cosmeceutical and pharmaceutical applications for AGA patients.

Differences in biochemical, physiological and molecular response mechanisms of rice, weedy rice and barnyardgrass subjected to drought

ABSTRACT The drought stress tolerance may differ among species and cultivars. The drought effect on plants depends on the impact on the plant’s physiological, biochemical and molecular processes, as well as on its ability to adapt under these conditions. This study aimed to evaluate the physiological and biochemical responses, and determine the expression of the genes OsAPX2, OsHSP24.15, OsHSP71.10 and OsHSP85.88 under drought conditions in rice, weedy rice and barnyardgrass. A greenhouse experiment was performed in a complete randomized design, with four replications and two factors: water conditions (well-watered and water deficit) and plant species [rice (Oryza sativa cv. Puitá), weedy rice (Oryza spp.) and barnyardgrass (Echinochloa spp.)]. Under drought conditions, the rice and weedy rice plants showed more cellular damage than the barnyardgrass, and the three species showed a reduced photosynthetic rate. C3 plants (rice and weedy rice) increased the damage to lipids and proteins at 5 days of drought. However, for C4 plants (barnyardgrass), the drought conditions did not affect the biochemical parameters. The expression of the OsHSP85.88 gene increased in the three plants exposed to water deficit.

INTERACTION OF FOLIAR NUTRIENTS AND HERBICIDES INTO PRODUCTION

A field trial was carried out at the fields of Baghdad University - Jadriya for the first location during the summer season of 2018, while the second location was in Mishkab Rice Research Station, Al-Najaf al-ashraf governorate during the summer season of 2019. This study was aimed investigate rice (Oryza sativa cv. Anber) response to foliar application of nutrition with the use of different herbicides. The design was RCBD wihin using split plot arrangement four replicates. The nutrition treatments; (Humic acide, Seaweed, Nano fertilizer, Normal fertilization and Without fertilizer) were in main plots. Herbicides treatments (Ronstar, Oscar, Rainbow, Super flag, weed free and weedy) occupied the sub plots.The results indicated that Ronstar herbicide was superior in giving the lowest average weeds density value 6.55 and 9.60 plant m-2 respectively, and highest average of grain yield was 2.53 and 3.42 t.h-1, respectively. The normal fertilization treatment was superior gave the highest average grain yield value 3.07 and 4.13 t h-1. As for the effects of interaction, the Ronstar combination with normal fertilization gave highest grain yield value 3.55 and 4.59 t h-1, respectively. Concluded from this trial that the Ronstar herbicide is the best among the herbicides used with different fertilization treatments by achieving the highest averages with most of the studied charactristics.

Structural analysis of regulatory stress elements in OsCIPK14 the rice homologue and functional validation in wheat

A major mechanism of stress tolerance in plants is to increase the cytosolic concentration of Ca2+ ion, a secondary messenger that mediates cellular responsiveness to the extracellular environment. The involvement of different types of Ca2+-sensing protein kinases such as CIPK in stress responses has been elucidated in different plant models. In this study, the upstream regulatory sequence of a CIPK homologue from an Egyptian rice cultivar (Oryza sativa cv Giza182) was structurally and functionally characterized and tested for its ability to drive the expression of β- glucuronidase reporter gene (uidA) in wheat. To that end, a 1275-bp DNA fragment, 49 nucleotides upstream of the putative transcription start site (TSS) of the OsCIPK14 coding sequence, referred in this study as URS-CIPK14, was cloned, sequenced and analyzed for the presence of stress-responsiveness cis-acting elements. Subsequently, the URS-CIPK14 was fused to the uidA gene with and without the Act-1 intron, cloned in pAB6 and transformed into wheat mature embryo for transient expression. Our results showed that URS-OsCIPK14 fused with the Act-1 intron was induced by NaCl and ABA, whereas URS-OsCIPK14 drove expression of uidA constitutively in the absence of Act-1 intron. Furthermore, rice wide-genome screening using URS-CIPK14 sequence obtained in this study identified another homologue on chromosome 11. Unlike URS-CIPK14, which is associated with chromosome 12, this homologue located at about 1015 nucleotides upstream from the putative TSS of OsCIPK15 coding sequence. Interestingly, the 5’-UTS in CIPK14 is longer than CIPK15 and can code for a putative short ORF that contains two Casein kinase II phosphorylation motifs (SXXD). The significance of this ORF is not known at this time and needs to be elucidated in future studies. We conclude that data from this work suggest that URS-CIPK14 can be used as a promoter candidate for high expression level under both constitutive and inducible conditions.