Summary
The CRISPR/Cas9 system has been demonstrated to efficiently induce targeted gene editing in a variety of organisms including plants. Recent work showed that CRISPR/Cas9‐induced gene mutations in Arabidopsis were mostly somatic mutations in the early generation, although some mutations could be stably inherited in later generations. However, it remains unclear whether this system will work similarly in crops such as rice. In this study, we tested in two rice subspecies 11 target genes for their amenability to CRISPR/Cas9‐induced editing and determined the patterns, specificity and heritability of the gene modifications. Analysis of the genotypes and frequency of edited genes in the first generation of transformed plants (T0) showed that the CRISPR/Cas9 system was highly efficient in rice, with target genes edited in nearly half of the transformed embryogenic cells before their first cell division. Homozygotes of edited target genes were readily found in T0 plants. The gene mutations were passed to the next generation (T1) following classic Mendelian law, without any detectable new mutation or reversion. Even with extensive searches including whole genome resequencing, we could not find any evidence of large‐scale off‐targeting in rice for any of the many targets tested in this study. By specifically sequencing the putative off‐target sites of a large number of T0 plants, low‐frequency mutations were found in only one off‐target site where the sequence had 1‐bp difference from the intended target. Overall, the data in this study point to the CRISPR/Cas9 system being a powerful tool in crop genome engineering.
Highly porous polypyrrole (PPy)-coated TiO 2 /ZnO nanofibrous mat has been successfully synthesized. The core TiO 2 / ZnO nanofibers have an average diameter of ca. 100 nm and the shell of ultrathin PPy layer has a thickness of ca. 7 nm. The NH 3 gas sensor using the as-prepared material exhibited a fast response over a wide dynamic range and high sensitivity with a detection limit of 60 ppb (S/N ¼ 3). Compared to conventional pristine PPy film, the improved performance in NH 3 detection can be attributed to the free access of NH 3 to PPy and a minimized gas diffusion resistance through the ultrathin PPy layer.
SummaryPlant sense potential microbial pathogen using pattern recognition receptors (PRRs) to recognize pathogen‐associated molecular patterns (PAMPs). The Lectin receptor‐like kinase genes (LecRKs) are involved in various cellular processes mediated by signal transduction pathways. In the present study, an L‐type lectin receptor kinase gene LecRK‐V was cloned from Haynaldia villosa, a diploid wheat relative which is highly resistant to powdery mildew. The expression of LecRK‐V was rapidly up‐regulated by Bgt inoculation and chitin treatment. Its transcript level was higher in the leaves than in roots, culms, spikes and callus. Single‐cell transient overexpression of LecRK‐V led to decreased haustorium index in wheat variety Yangmai158, which is powdery mildew susceptible. Stable transformation LecRK‐V into Yangmai158 significantly enhanced the powdery mildew resistance at both seedling and adult stages. At seedling stage, the transgenic line was highly resistance to 18 of the tested 23 Bgt isolates, hypersensitive responses (HR) were observed for 22 Bgt isolates, and more ROS at the Bgt infection sites was accumulated. These indicated that LecRK‐V confers broad‐spectrum resistance to powdery mildew, and ROS and SA pathways contribute to the enhanced powdery mildew resistance in wheat.
mRNAs containing premature termination codons (PTCs) are known to be degraded via nonsense-mediated mRNA decay (NMD). Unexpectedly, we found that mRNAs containing any type of PTCs (UAA, UAG, and UGA) are detained in the nucleus, whereas their wild-type counterparts are rapidly exported. This retention is strictly reading-frame dependent. Strikingly, our data indicate that translating ribosomes in the nucleus proofread the frame and detect the PTCs in the nucleus. Moreover, the shuttling NMD protein Upf1 specifically associates with PTC+mRNAs (PTC-containing mRNAs) in the nucleus and is required for nuclear retention of PTC+mRNAs. Together, our data lead to a working model that PTCs are recognized in the nucleus by translating ribosomes, resulting in recruitment of Upf1, which in turn functions in nuclear retention of PTC+mRNA. Nuclear PTC recognition adds a new layer of proofreading for mRNA and may be vital for ensuring the extraordinary fidelity required for protein production.
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