Small RNA and Degradome Deep Sequencing Reveal Regulatory Roles of MicroRNAs in Response to Sugarcane Mosaic Virus Infection on Two Contrasting Sugarcane Cultivars.

MicroRNAs (miRNAs) play an essential regulatory role in plant-virus interaction. However, few studies have focused on the roles of miRNAs and their targets after sugarcane mosaic virus (SCMV) infection in sugarcane. To address this issue, we conducted small RNA (sRNA) and degradome sequencing on two contrasting sugarcanes (SCMV-resistant 'Fuoguo1' [FG1] and susceptible 'Badila') infected by SCMV at five time points. A total of 1,578 miRNAs were profiled from 30 sRNA libraries, comprising 660 known miRNAs and 380 novel miRNAs. Differential expression analysis of miRNAs revealed that most were highly expressed during the SCMV exponential phase in Badila at 18 h postinfection, with expression profiles positively correlated with virus replication dynamics as observed through clustering. Analysis of degradome data indicated a higher number of differential miRNA targets in Badila compared to FG1 at 18 h postinfection. Gene ontology (GO) enrichment analysis significantly enriched the stimulus-response pathway, suggesting negative regulatory roles to SCMV resistance. Specifically, miR160 upregulated expression patterns and validated in Badila through quantitative real-time PCR (qRT-PCR) in the early stages of SCMV multiplication. Our research provides new insights into the dynamic response of plant miRNA and virus replication and contributes valuable information on the intricate interplay between miRNAs and SCMV infection dynamics. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

High-Quality Genome Sequence Resource for Fusarium andiyazi Causing Pokkah Boeng Disease of Sugarcane in China.

Sugarcane pokkah boeng disease (PBD) is emerging as a prevalent foliar disease in China. This airborne disease is caused by the Fusarium species complex. To investigate the diversity and evolution of Fusarium spp., we performed whole-genome sequencing of Fusarium andiyazi YN28 using a combination of Oxford Nanopore and Illumina technology. The F. andiyazi YN28 genome was sequenced, assembled, and annotated. A high-quality genome was assembled into 24 contigs with an N50 of 2.80 Mb. The genome assembly generated a total size of 44.1 Mb with a GC content of 47.64%. In total, 15,508 genes were predicted, including 794 genes related to the carbohydrate-active enzymes, 397 noncoding RNA, 155 genes associated with transporter classification, 4,550 genes linked to pathogen-host interactions, and 269 genes involved in effector proteins. Collectively, our results will provide insight into the host-pathogen interactions and will facilitate the breeding of new varieties of sugarcane resistant to PBD.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Characterization of CBL-CIPK signaling networks and their response to abiotic stress in sugarcane.

Calcineurin B-like proteins (CBLs) perceive calcium signals triggered by abiotic stress and interact with CBL-interacting protein kinases (CIPKs) to form a complex signal network. This study identified 21 SsCBL and 89 SsCIPK genes in Saccharum spontaneum, and 90 ScCBL and 367 ScCIPK genes in the sugarcane cultivar ZZ1. Phylogenetic analysis classified CBL genes into three groups and CIPK genes into twenty-five groups, with whole-genome duplication events promoting their expansion in sugarcane. RNA-seq analysis revealed their involvement in abiotic stress responses through ABA, JA, and SA pathways. Four ScCBLs and eight ScCIPKs were cloned from ZZ1. Three CBL-CIPK interactions were detected using a yeast two-hybrid system and Firefly luciferase complementation imaging, showing CBLs as membrane proteins and CIPKs as nuclear proteins. Spatial expression profiles indicate these genes are expressed in various tissues, with the highest expression in roots. Gene expression analyses suggested that CBL-CIPK signaling networks are involved in responses to drought, salt, and reactive oxygen species, possibly through Ca2+-induced hormone pathways. These findings establish three CBL-CIPK signaling networks responding to abiotic stress, providing a molecular basis for improving sugarcane stress resistance.

Production of porcine aminopeptidase N (pAPN) site-specific edited pigs.

Porcine viral diarrhea is an acute and highly contagious enteric disease in pigs which causes huge economic losses in pig industry worldwide. Transmissible gastroenteritis virus (TGEV) is main pathogens responsible for piglets viral diarrhea. Knockout the host cellular surface receptor for TGEV may be an effective way to accelerate the breeding of resistant pigs. In this study, we applied site-specific editing pAPN which is effective in swine testis (ST) cells. Site-specific editing of pAPN reduced TGEV proliferation in ST cells by 96%-99% at different time periods post-infection. Next, the site-specific editing of pAPN porcine fetal fibroblasts were produced, and then the cell colonies were used as donor cells to generate the site-specific editing of pAPN pigs. Our research findings will not only offer a more thorough understanding of the pathogenesis of piglet diarrhea and lay the foundation for breeding TGEV-resistant piglets, but also understanding the molecular mechanisms involved in coronaviral infections.