Disruption of Abcc6 Transporter in Zebrafish Causes Ocular Calcification and Cardiac Fibrosis

Pseudoxanthoma elasticum (PXE), caused by ABCC6/MRP6 mutation, is a heritable multisystem disorder in humans. The progressive clinical manifestations of PXE are accompanied by ectopic mineralization in various connective tissues. However, the pathomechanisms underlying the PXE multisystem disorder remains obscure, and effective treatment is currently available. In this study, we generated zebrafish abcc6a mutants using the transcription activator-like effector nuclease (TALEN) technique. In young adult zebrafish, abcc6a is expressed in the eyes, heart, intestine, and other tissues. abcc6a mutants exhibit extensive calcification in the ocular sclera and Bruch’s membrane, recapitulating part of the PXE manifestations. Mutations in abcc6a upregulate extracellular matrix (ECM) genes, leading to fibrotic heart with reduced cardiomyocyte number. We found that abcc6a mutation reduced levels of both vitamin K and pyrophosphate (PPi) in the serum and diverse tissues. Vitamin K administration increased the gamma-glutamyl carboxylated form of matrix gla protein (cMGP), alleviating ectopic calcification and fibrosis in vertebrae, eyes, and hearts. Our findings contribute to a comprehensive understanding of PXE pathophysiology from zebrafish models

Tillage and straw mulching impacts on grain yield and water use efficiency of spring maize in Northern Huang–Huai–Hai Valley

A two-year field experiment (2012–2013) was conducted to investigate the effects of two tillage methods and five maize straw mulching patterns on the yield, water consumption, and water use efficiency (WUE) of spring maize (Zea mays L.) in the northern Huang–Huai–Hai valley of China. Compared to rotary tillage, subsoil tillage resulted in decreases in water consumption by 6.3–7.8% and increases in maize yield by 644.5–673.9 kg ha− 1, soil water content by 2.9–3.0%, and WUE by 12.7–15.2%. Chopped straw mulching led to higher yield, soil water content, and WUE as well as lower water consumption than prostrate whole straw mulching. Mulching with 50% chopped straw had the largest positive effects on maize yield, soil water content, and WUE among the five mulching treatments. Tillage had greater influence on maize yield than straw mulching, whereas straw mulching had greater influence on soil water content, water consumption, and WUE than tillage. These results suggest that 50% chopped straw mulching with subsoil tillage is beneficial in spring maize production aiming at high yield and high WUE in the Huang–Huai–Hai valley

Systematic genome editing of the genes on zebrafish Chromosome 1 by CRISPR/Cas9

Genome editing by the well-established CRISPR/Cas9 technology has greatly facilitated our understanding of many biological processes. However, a complete whole-genome knockout for any species or model organism has rarely been achieved. Here, we performed a systematic knockout of all the genes (1333) on Chromosome 1 in zebrafish, successfully mutated 1029 genes, and generated 1039 germline-transmissible alleles corresponding to 636 genes. Meanwhile, by high-throughput bioinformatics analysis, we found that sequence features play pivotal roles in effective gRNA targeting at specific genes of interest, while the success rate of gene targeting positively correlates with GC content of the target sites. Moreover, we found that nearly one-fourth of all mutants are related to human diseases, and several representative CRISPR/Cas9-generated mutants are described here. Furthermore, we tried to identify the underlying mechanisms leading to distinct phenotypes between genetic mutants and antisense morpholino-mediated knockdown embryos. Altogether, this work has generated the first chromosome-wide collection of zebrafish genetic mutants by the CRISPR/Cas9 technology, which will serve as a valuable resource for the community, and our bioinformatics analysis also provides some useful guidance to design gene-specific gRNAs for successful gene editing