The outbreak of the COVID-19 pandemic is partially due to the challenge of identifying asymptomatic and pre-symptomatic carriers of the virus, and thus highlights a strong motivation for diagnostics that can be rapidly deployed with high sensitivity. On the other hand, several concerned SARS-CoV-2 variants, including the Omicron, are required to be identified as soon as the samples are identified as ‘positive’. Unfortunately, a traditional PCR test does not allow their specific identification. Herein, for the first time, we have developed MOPCS (Methodologies of Photonic CRISPR Sensing), which combines an optical sensing technology-surface plasmon resonance (SPR), and the ‘gene scissors’ CRISPR technique to achieve both high sensitivity and specificity of viral variants’ measurement. MOPCS is a low-cost, CRISPR/Cas12a system-empowered SPR gene detecting platform that can analyze viral RNA, without the need for amplification, within 38 min from sample input to results output, and achieve a limit of detection of 15 fM. MOPCS achieves a highly sensitive analysis of SARS-CoV-2 and mutations appear in variants B.1.617.2 (Delta), B.1.1.529 (Omicron), and BA.1 (a subtype of Omicron). This platform was also used to analyze some recently collected patient samples from a local outbreak in China and identified by the Centers for Disease Control and Prevention. This innovative CRISPR-empowered SPR platform will further contribute to various fast, sensitive, and accurate detection of target nucleic acid sequences with single-base mutations.

In this paper, the influence of drought hardening on the growth, development, resistance physiology, leaf microstructure and stomatal behavior of potato seedlings under drought stress was studied, and the mechanism of drought hardening improvement of potato seedling drought resistance was elucidated. We found that drought stress had several adverse effects on potato seedlings, yet drought hardening alleviated the decrease in relative water content (RWC), net photosynthetic rate (P n) and chlorophyll content and inhibited the increase in relative electric conductivity and malondialdehyde (MDA) content. Compared with contrast seedlings, drought-hardened seedlings also had enhanced root vigor, increased antioxidant enzyme activity and higher levels of abscisic acid (ABA), proline (Pro), soluble sugars and polyamines (PAs) under drought stress. In addition, the stomatal density of potato seedling leaves increased significantly, while the leaf area, stomatal size and stomatal aperture decreased with drought hardening treatment. These changes led to reduced leaf transpiration rate (T r) and improved water utilization efficiency (WUE). The changes in leaf microstructure also had a positive effect on the drought resistance of the drought-hardened potato seedlings. So it can be concluded that through increasing the content of some endogenous hormones, osmotic regulatory substances and the activities of antioxidant enzymes, the resistance physiology of drought-hardened potato seedlings was enhanced.

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Coronavirus disease 2019 (COVID-19) is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The gold standard method for the diagnosis of SARS-CoV-2 depends on quantitative reverse transcription-polymerase chain reaction till now, which is time-consuming and requires expensive instrumentation, and the confirmation of variants relies on further sequencing techniques. Herein, we first proposed a robust technique-methodology of electrochemical CRISPR sensing with the advantages of rapid, highly sensitivity and specificity for the detection of SARS-CoV-2 variant. To enhance the sensing capability, gold electrodes are uniformly decorated with electro-deposited gold nanoparticles. Using DNA template identical to SARS-CoV-2 Delta spike gene sequence as model, our biosensor exhibits excellent analytical detection limit (50 fM) and high linearity (R2 = 0.987) over six orders of magnitude dynamic range from 100 fM to 10 nM without any nucleic-acid-amplification assays. The detection can be completed within 1 h with high stability and specificity which benefits from the CRISPR-Cas system. Furthermore, based on the wireless micro-electrochemical platform, the proposed biosensor reveals promising application ability in point-of-care testing.

Since the end of 2019, a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has deprived numerous lives worldwide, called COVID-19. Up to date, omicron is the latest variant of concern, and BA.5 is replacing the BA.2 variant to become the main subtype rampaging worldwide. These subtypes harbor an L452R mutation, which increases their transmissibility among vaccinated people. Current methods for identifying SARS-CoV-2 variants are mainly based on polymerase chain reaction (PCR) followed by gene sequencing, making time-consuming processes and expensive instrumentation indispensable. In this study, we developed a rapid and ultrasensitive electrochemical biosensor to achieve the goals of high sensitivity, the ability of distinguishing the variants, and the direct detection of RNAs from viruses simultaneously. We used electrodes made of MXene-AuNP (gold nanoparticle) composites for improved sensitivity and the CRISPR/Cas13a system for high specificity in detecting the single-base L452R mutation in RNAs and clinical samples. Our biosensor will be an excellent supplement to the RT-qPCR method enabling the early diagnosis and quick distinguishment of SARS-CoV-2 Omicron BA.5 and BA.2 variants and more potential variants that might arise in the future.

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WD40 proteins, also known as WD40 domain proteins, constitute a large gene family in eukaryotes and play multiple roles in cellular processes. However, systematic identification and analysis of WD40 proteins have not yet been reported in potato (Solanum tuberosum L.). In the present study, 178 potato WD40 (StWD40) genes were identified and their distribution on chromosomes, gene structure, and conserved motifs were assessed. According to their structural and phylogenetic protein features, these 178 StWD40 genes were classified into 14 clusters and 10 subfamilies. Collinearity analysis showed that segmental duplication events played a major role in the expansion of the StWD40 gene family. Synteny analysis indicated that 45 and 23 pairs of StWD40 genes were orthologous to Arabidopsis and wheat (Triticum aestivum), respectively, and that these gene pairs evolved under strong purifying selection. RNA-seq data from different tissues and abiotic stresses revealed tissue-specific expression and abiotic stress-responsive StWD40 genes in doubled monoploid potato (DM). Furthermore, we further analyzed the WD40 genes might be involved in anthocyanin biosynthesis and drought stress in tetraploid potato cultivars based on RNA-seq data. In addition, a protein interaction network of two homologs of Arabidopsis TTG1, which is involved in anthocyanin biosynthesis, was constructed to identify proteins that might be related to anthocyanin biosynthesis. The result showed that there were 112 pairs of proteins interacting with TTG1, with 27 being differentially expressed in pigmented tissues. This study indicates that WD40 proteins in potato might be related to anthocyanin biosynthesis and abiotic stress responses.

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