The Function of DNA Demethylase Gene ROS1a Null Mutant on Seed Development in Rice (Oryza Sativa) Using the CRISPR/CAS9 System

The endosperm is the main nutrient source in cereals for humans, as it is a highly specialized storage organ for starch, lipids, and proteins, and plays an essential role in seed growth and development. Active DNA demethylation regulates plant developmental processes and is ensured by cytosine methylation (5-meC) DNA glycosylase enzymes. To find out the role of OsROS1a in seed development, the null mutant of OsROS1a was generated using the CRISPR/Cas9 system. The null mutant of OsROS1a was stable and heritable, which affects the major agronomic traits, particularly in rice seeds. The null mutant of OsROS1a showed longer and narrower grains, and seeds were deformed containing an underdeveloped and less-starch-producing endosperm with slightly irregularly shaped embryos. In contrast to the transparent grains of the wild type, the grains of the null mutant of OsROS1a were slightly opaque and rounded starch granules, with uneven shapes, sizes, and surfaces. A total of 723 differential expression genes (DEGs) were detected in the null mutant of OsROS1a by RNA-Seq, of which 290 were downregulated and 433 were upregulated. The gene ontology (GO) terms with the top 20 enrichment factors were visualized for cellular components, biological processes, and molecular functions. The key genes that are enriched for these GO terms include starch synthesis genes (OsSSIIa and OsSSIIIa) and cellulose synthesis genes (CESA2, CESA3, CESA6, and CESA8). Genes encoding polysaccharides and glutelin were found to be downregulated in the mutant endosperm. The glutelins were further verified by SDS-PAGE, suggesting that glutelin genes could be involved in the null mutant of OsROS1a seed phenotype and OsROS1a could have the key role in the regulation of glutelins. Furthermore, 378 differentially alternative splicing (AS) genes were identified in the null mutant of OsROS1a, suggesting that the OsROS1a gene has an impact on AS events. Our findings indicated that the function on rice endosperm development in the null mutant of OsROS1a could be influenced through regulating gene expression and AS, which could provide the base to properly understand the molecular mechanism related to the OsROS1a gene in the regulation of rice seed development.     

A Defect Tolerance Scheme for Nanotechnology Circuits

Lithography-based integrated circuit fabrication is rapidly approaching its limit in terms of feature size. The current alternative is nanotechnology-based fabrication, which relies on self-assembly of nanotubes or nanowires. Such a process is subject to a high defect rate, which can be tolerated using carefully crafted defect tolerance techniques. This paper presents an algorithm for reconfiguration-based defect tolerance in nanotechnology switches. The algorithm offers an average switch density improvement of 50% to 100% to most recently published techniques. The algorithm is also consistent in improving the yield through minimizing false rejects as the results show over a large sample. The improvement percentage varies depending on the manufactured switch size and the desired defect-free size with the improvement in efficiency directly proportional to the size of the switch.     

Molecular Finger Printing of Nutra-Coconut Oil with Improved Health Protective Phytoceuticals and its Efficacy as Frying Medium

Coconut oil is rich in medium chain triglycerides but lacks polyunsaturated fatty acids (PUFA) and bio‐active phytoceuticals. In the present work nutra‐coconut oil was prepared by blending coconut oil and flaxseed oil (70:30) and adding 3000 ppm of flaxseed cake concentrate using ethanol, methanol and 20 % aqueous ethanol. The concentrate prepared from flaxseed was from ethanol as it gave maximum yield. The different bio‐active molecules in flaxseed concentrate observed are polyphenols (39.04 %), tocopherols (4.37 %), ferulic acid (0.17 mg g^?1), p‐coumaric acid (2.24 mg g^?1), chlorogenic acid (16.11 mg g^?1), gallic acid (8.58 mg g^?1), sinapic acid (0.64 mg g^?1) and secoisolariresinol (30.13 mg g^?1). The nutra‐coconut oil was found to have polyphenols (2.86 %), tocopherols (442.96 ppm) and antiradical activity (94 %). The PUFA content was found to increase in nutra‐coconut oil significantly (p < 0.05) (2–22 %). The FT‐IR spectra of nutra‐coconut oil revealed that the peak at 3009 and 1651 cm^?1 was associated with the presence of unsaturated fatty acids. There was no significant (p > 0.05) difference observed in sensory attributes of snack food fried using coconut oil and nutra‐coconut oil indicating that the later could be used as a frying medium and useful for food processing industries.     

RF plasma-polymerization of acetylene: Correlation between plasma diagnostics and deposit characteristics

Clay-polymer nanocomposites are developed to obtain better mechanical, electrical or permeability properties controlled by the interaction energy between polymer chains and filler dispersed in polymeric blend. The dispersion of clay particles in polymers was achieved by chemical pre-treatment of the individual clay layers. The technique of acetylene plasma deposition is proposed as an alternative route to alter hydrophilic character of the clay. Since the clay is a complex material, plasma-polymer, coated silicon wafer was used as reference and characterized by different surface techniques. Different plasma parameters were chosen to lead to the most hydrophobic coating. Optical Emission Spectroscopy (OES) was also used to establish the relationship between the plasma chemistry and the properties of the deposit layer.     

Test Set Compression Through Alternation Between Deterministic and Pseudorandom Test Patterns

This paper presents a new reseeding technique that reduces the storage required for the seeds as well as the test application time by alternating between ATPG and reseeding to optimize the seed selection. The technique avoids loading a new seed into the PRPG whenever the PRPG can be placed in a state that generates test patterns without explicitly loading a seed. The ATPG process is tuned to target only undetected faults as the PRPG goes through its natural sequence which is maximally used to generate useful test patterns. The test application procedure is slightly modified to enable higher flexibility and more reduction in tester storage and test time. The results of applying the technique show up to 90% reduction in tester storage and 80% reduction in test time compared to classic reseeding. They also show 70% improvement in defect coverage when the technique is emulated on test chips with real defects.     

Characterisation and comparison of the micromixing efficiency in torus and batch stirred reactors

The mixing at a molecular scale (micromixing) plays an important role on selectivity, yield and quality of final products of a large range of competing fast chemical reactions. In this study, we have compared, by the use of iodide–iodate reaction tests, the micromixing in two reactors, the first one is the standard batch stirred reactor and the second is the torus reactor. Various conditions of agitation and feed locations were used for this study. A comparative analysis of the micromixedness ratio (α) in the two reactors was carried out on the basis of the local rate of specific energy dissipation.     

Agarose Hydrogel Beads: An Effective Approach to Improve the Catalytic Activity,Stability and Reusability of Fungal Amyloglucosidase of GH15 Family

This study deals with the immobilization of amyloglucosidase within agarose using method of entrapment. Enzyme was produced from Aspergillus fumigatus KIBGE-IB33 and then partially purified using 40% ammonium sulphate saturation. Using 40 gl^?1 concentration of agarose and adjusting 3.0 mm size of hydrogels, maximum entrapment yield (78%) was obtained. The kinetic behavior was slightly changed after immobilization as reaction time and reaction temperature increases from 5.0 min (soluble) to 10.0 min (immobilized) and 60 °C (soluble) to 65 °C (immobilized), respectively while, pH optima remained same (pH 5.0). Substrate saturation kinetics revealed that K_m was increased from 1.47 to 4.215 mg ml^?1 while, the value of V_max decreased from 947 to 611 kU mg^?1 for soluble and entrapped amyloglucosidase, respectively. The stability profile of amyloglucosidase also significantly improved after entrapment in agarose hydrogels at 50, 60 and 70 °C for 120 min with retention of 77, 59 and 25% residual activity, respectively. Furthermore, the t_1/2 of soluble and immobilized amyloglucosidase at 60 °C was 167 and 375 min respectively. Due to increase in reusability for various subsequent cycles of entrapped amyloglucosidase, about 8.73 mg ml^?1 increase in glucose production was observed as compared to soluble enzyme.

Graphical Abstract

Open image in new window