CRISPR/Cas9-Mediated Gene Editing in Grain Crops

The development of reliable and efficient techniques for making precise targeted changes in the genome of living organisms has been a long-standing objective of researchers throughout the world. In plants, different methods, each with several different variations, have been developed for this purpose, though many of them are hampered either by providing only temporary modification of gene function or unpredictable off-target results. The recent discovery of clustered regularly interspaced short palindromic repeats (CRISPRs) and the CRISPR-associated 9 (Cas9) nucleases started a new era in genome editing. Basically, the CRISPR/Cas system is a natural immune response of prokaryotes to resist foreign genetic elements entering via plasmids and phages. Through this naturally occurring gene editing system, bacteria create DNA segments known as CRISPR arrays that allow them to “remember” foreign genetic material for protection against it and other similar sequences in the future. This system has now been adopted by researchers in laboratory to create a short guide RNA that binds to specific target sequences of DNA in eukaryotic genome, and the Cas9 enzyme cuts the DNA at the targeted location. Once cut, the cell’s endogenous DNA repair machinery is used to add, delete, or replace pieces of genetic material. Though CRISPR/Cas9 technology has been recently developed, it has started to be regularly used for gene editing in plants as well as animals to good success. It has been proved as an efficient transgene-free technique. A simple search on PubMed (NCBI) shows that among all plants, 80 different studies published since 2013 involved CRISPR/Cas9-mediated genome editing in rice. Of these, 20, 13, and 24 papers have been published in 2019, 2018, and 2017, respectively. Furthermore, 20 different studies published since 2014 utilized CRISPR/Cas9 system for gene editing in wheat, where five of these studies were published in 2019 and seven were published in 2018. Genomes of other grain crops edited through this technique include maize, sorghum, barley, etc. This indicates the high utility of this technique for gene editing in grain crops. Here we emphasize on CRISPR/Cas9-mediated gene editing in rice, wheat, and maize

Temperature dependence of Mott transition in VO_2 and programmable critical temperature sensor

The temperature dependence of the Mott metal-insulator transition (MIT) is studied with a VO_2-based two-terminal device. When a constant voltage is applied to the device, an abrupt current jump is observed with temperature. With increasing applied voltages, the transition temperature of the MIT current jump decreases. We find a monoclinic and electronically correlated metal (MCM) phase between the abrupt current jump and the structural phase transition (SPT). After the transition from insulator to metal, a linear increase in current (or conductivity) is shown with temperature until the current becomes a constant maximum value above T_{SPT}=68^oC. The SPT is confirmed by micro-Raman spectroscopy measurements. Optical microscopy analysis reveals the absence of the local current path in micro scale in the VO_2 device. The current uniformly flows throughout the surface of the VO_2 film when the MIT occurs. This device can be used as a programmable critical temperature sensor.Comment: 4 pages, 3 figure

Development of a Semirigid Pavement Incorporating Ultrarapid Hardening Cement and Chemical Admixtures for Cement Grouts

Mechanical tests were carried out to evaluate the influence and effects of fluidity and compressive strength of cement grout on semirigid asphalt pavement. An open graded asphalt skeleton was designed in order to achieve target porosity in the range of 18~22%. In addition, four types of cement grout mixtures were produced with varying mix proportions with ultrarapid hardening cement and chemical admixtures, that is, accelerating and retarding agents. For the semirigid pavement specimens, mechanical experiments to measure properties such as porosity, flexural strength, Marshall stability, and wheel tracking resistance were carried out. The test results demonstrated that the flow time (fluidity) of cement grout is the most significant factor that determines the mechanical properties of semirigid asphalt specimens under constant condition of the open graded asphalt skeleton. For the semirigid pavement mixing proportion in the current study, it is recommended that the porosity of the open graded asphalt skeleton and flow time of cement grout should be 20% and within 12 seconds, respectively

Performance Analysis of CFRP Composite Strips Confined RC Columns under Axial Compression

In an attempt to mitigate the high cost of FRP composite strengthening, an experimental investigation was carried out that sought to achieve efficient and most favorable FRP strengthening using CFRP composite strips. 50 mm wide CFRP composite strips were used in two different spacings (20 mm and 40 mm) to confine columns. The test results of the column confined with smaller spacing (20 mm) showed significant restraint of axial deformation of the column and enhanced the strength capacity to a maximum of 99.20% compared to that of reference column. In contrast, the column confined by strips with larger spacing (40 mm) failed by crushing of concrete alone, which occurred even before the CFRP strips reached their ultimate strain. In addition, the embodied energy that exists in the CFRP strips could not be utilized effectively. The stress and strength enhancement ratio of this present study was compared with the previous research that has been conducted on columns confined with full wrapping. From the obtained results, it is recommended that CFRP strips with a spacing of 20 mm be used to improve the strength capacity of the RC column; in addition, this wrapping technique provides economic benefits compared to a column confined with full wrapping

A potential role of coumestrol in soybean leaf senescence and its interaction with phytohormones

Coumestrol is a natural organic compound synthesized in soy leaves and functions as a phytoalexin. The coumestrol levels in plants are reported to increase upon insect attack. This study investigates the correlation between coumestrol, senescence, and the effect of phytohormones on the coumestrol levels in soybean leaves. Our analysis involving high-performance liquid chromatography and 2-D gel electrophoresis indicated a significant difference in the biochemical composition of soybean leaves at various young and mature growth stages. Eight chemical compounds were specifically detected in young leaves (V1) only, whereas three different coumestans isotrifoliol, coumestrol, and phaseol were detected only in mature, yellow leaves of the R6 and R7 growth stage. MALDI-TOF-MS analysis was used to identify two proteins 3,9 -dihydroxypterocarpan 6A-monooxygenase (CYP93A1) and isoflavone reductase homolog 2 (IFR2) only in mature leaves, which are key components of the coumestrol biosynthetic pathway. This indicates that senescence in soybean is linked to the accumulation of coumestrol. Following the external application of coumestrol, the detached V1-stage young leaves turned yellow and showed an interesting development of roots at the base of the midrib. Additionally, the application of phytohormones, including SA, methyl jasmonate (MeJA), and ethephon alone and in various combinations induced yellowing within 5 days of the application with a concomitant significant increase in endogenous coumestrol accumulation. This was also accompanied by a significant increase in the expression of genes CYP81E28 (Gm08G089500), CYP81E22 (Gm16G149300), GmIFS1, and GmIFS2. These results indicate that various coumestans, especially coumestrol, accumulate during leaf maturity, or senescence in soybean