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Yanling Yin, Kezhen Qin, Xuewei Song, Qihao Zhang, Yanhong Zhou, Xiaojian Xia, Jingquan Yu
BRASSINAZOLE RESISTANT 1 (BZR1), the critical regulator of brassinosteroid (BR) response, participates in various BR-mediated developmental processes. However, the roles of BZR1 in stress tolerance are less clear. Here, we found that BZR1-like protein in tomato controls BR response and is involved in thermotolerance by regulating the FERONIA (FER) homologs. The CRISPR-bzr1 mutant showed reduced growth and was not responsive to 24-epibrassinolide (EBR) as regard to the promotion of plant growth. Mutation in BZR1 impaired the induction of RESPIRATORY BURST OXIDASE HOMOLOG1 (RBOH1), production of H2O2 in the apoplast, and heat tolerance...
August 13, 2018: Plant & Cell Physiology
Yoko Ikeda, Ryuichi Nishihama, Shohei Yamaoka, Mario A Arteaga-Vazquez, Adolfo Aguilar-Cruz, Daniel Grimanelli, Romain Pogorelcnik, Robert A Martienssen, Katsuyuki T Yamato, Takayuki Kohchi, Takashi Hirayama, Olivier Mathieu
DNA methylation is an epigenetic mark that ensures silencing of transposable elements (TEs) and affects gene expression in many organisms. The function of different DNA methylation regulatory pathways has been largely characterized in the model plant Arabidopsis thaliana. However, far less is known about DNA methylation regulation and functions in basal land plants. Here we focus on the liverwort Marchantia polymorpha, an emerging model species that represents a basal lineage of land plants. We identified MpMET, the M...
August 9, 2018: Plant & Cell Physiology
Luan L Chu, Dipesh Dhakal, Hee J Shin, Hye J Jung, Tokutaro Yamaguchi, Jae K Sohng
Flavonoids are one of the predominant groups of plant polyphenols, and these compounds have significant effects on human health and nutrition. Sulfated flavonoids have more favorable attributes compared to their parent compounds such as increased solubility, stability, and bioavailability. In this research, we developed a microbial system to produce sulfated naringenin using Escherichia coli expressing a sulfotransferase (ST) from Arabidopsis thaliana (At2g03770). This wild-type strain was used as a model system for testing clustered regularly interspaced short palindromic repeats (CRISPR) interference (CRISPRi) metabolic engineering strategies...
2018: Frontiers in Microbiology
Rui Li, Lixing Zhang, Liu Wang, Lin Chen, Ruirui Zhao, Jiping Sheng, Lin Shen
Chilling stress is the main constraint in tomato (Solanum lycopersicum) production, as this is a chilling-sensitive horticultural crop. The highly conserved C-repeat binding factors (CBFs) are cold-response system components found in many species. In this study, we generated slcbf1 mutants using the CRISPR/Cas9 system and investigated the role of SlCBF1 in tomato plant chilling tolerances. The slcbf1 mutants exhibited more severe chilling injury symptoms with higher electrolyte leakage and malondialdehyde levels than wild type (WT) plants...
August 10, 2018: Journal of Agricultural and Food Chemistry
Shengchen Shan, Evgeny V Mavrodiev, Riqing Li, Zhengzhi Zhang, Bernard A Hauser, Pamela S Soltis, Douglas E Soltis, Bing Yang
Tragopogon (Asteraceae) is an excellent natural system for studies of recent polyploidy. Development of an efficient CRISPR/Cas9-based genome editing platform in Tragopogon will facilitate novel studies of the genetic consequences of polyploidy. Here, we report our initial results of developing CRISPR/Cas9 in Tragopogon. We have established a feasible tissue culture and transformation protocol for Tragopogon. Through protoplast transient assays, use of the TragCRISPR system (i.e. the CRISPR/Cas9 system adapted for Tragopogon) was capable of introducing site-specific mutations in Tragopogon protoplasts...
August 7, 2018: Molecular Ecology Resources
Michelle M C Buckner, Maria Laura Ciusa, Laura J V Piddock
Antimicrobial resistance (AMR) is a global problem hindering treatment of bacterial infections, rendering many aspects of modern medicine less effective. AMR genes (ARGs), are frequently located on plasmids, which are self-replicating elements of DNA. They are often transmissible between bacteria, and some have spread globally. Novel strategies to combat AMR are needed, and plasmid curing and anti-plasmid approaches could reduce ARG prevalence, and sensitize bacteria to antibiotics. We discuss the use of curing agents as laboratory tools including chemicals (e...
July 30, 2018: FEMS Microbiology Reviews
Rui Wu, Miriam Lucke, Yun-Ting Jang, Wangsheng Zhu, Efthymia Symeonidi, Congmao Wang, Joffrey Fitz, Wanyan Xi, Rebecca Schwab, Detlef Weigel
Background: Our knowledge of natural genetic variation is increasing at an extremely rapid pace, affording an opportunity to come to a much richer understanding of how effects of specific genes are dependent on the genetic background. To achieve a systematic understanding of such GxG interactions, it is desirable to develop genome editing tools that can be rapidly deployed across many different genetic varieties. Results: We present an efficient CRISPR/Cas9 toolbox of super module (SM) vectors...
2018: Plant Methods
Fanqian Yin, Wenfu Liu, Jing Chai, Bin Lu, Robert W Murphy, Jing Luo
Polyploidization occurs widely in eukaryotes, and especially in plants. Polyploid plants and some fishes have been commercialized. Typically, severe genomic perturbations immediately follow polyploidization and little is known about how polyploid offspring survives the genetic and epigenetic changes. Investigations into this require the identification of genes related to polyploidization and the discrimination of dosage-balance from paternal and maternal copies, and regardless of the mechanism being either autopolyploidization or allopolyploidization...
2018: Frontiers in Genetics
Rizka Rahmana Putri, Liangbiao Chen
CRISPR activation (CRISPRa) system is the convenient tool for targeted-gene activation, it has been developed and combined with a lighting-based system that can control transcription initiation spatially and temporally by utilizing photoreceptor derived from plant Arabidopsis thaliana. A blue light photoreceptor the Cryptochrome 2 (CRY2), and its binding partner CIB1 will dimerize by exposure to the blue light and it has been applied to human cells. However, the application of a combination of these two systems to zebrafish cell is still not explored...
August 1, 2018: Gene
Supriya B Aglawe, Kalyani M Barbadikar, Satendra K Mangrauthia, M Sheshu Madhav
Crop improvement is a continuous process in agriculture which ensures ample supply of food, fodder and fiber to burgeoning world population. Despite tremendous success in plant breeding and transgenesis to improve the yield-related traits, there have been several limitations primarily with the specificity in genetic modifications and incompatibility of host species. Because of this, new breeding techniques (NBTs) are gaining worldwide attention for crop improvement programs. Among the NBTs, genome editing (GE) using site-directed nucleases (SDNs) is an important and potential technique that overcomes limitations associated with classical breeding and transgenesis...
August 2018: 3 Biotech
Deepa Jaganathan, Karthikeyan Ramasamy, Gothandapani Sellamuthu, Shilpha Jayabalan, Gayatri Venkataraman
The availability of genome sequences for several crops and advances in genome editing approaches has opened up possibilities to breed for almost any given desirable trait. Advancements in genome editing technologies such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) has made it possible for molecular biologists to more precisely target any gene of interest. However, these methodologies are expensive and time-consuming as they involve complicated steps that require protein engineering...
2018: Frontiers in Plant Science
Ewen Callaway
No abstract text is available yet for this article.
August 2018: Nature
Penghui Xu, Hang Su, Wanli Chen, Pingli Lu
The CRISPR/Cas9 system has been widely used for generating targeted mutations in various species. In Arabidopsis, it largely relies on the edited cells where the Cas9 protein performs its activity to obtain heritable and stable mutated lines. Here, we designed an improved CRISPR/Cas9 system, named as the MSC (meiocyte-specific CRISPR/Cas9) system, in which the Cas9 expression is driven by an experimentally approved meiocyte-specific promoter ( AtDMC1 promoter). Two endogenous genes, including vegetative gene AtDET2 and reproductive gene AtDMC1 , were targeted...
2018: Frontiers in Plant Science
Yanglei Yi, Zhibo Li, Chunxu Song, Oscar P Kuipers
Bacillus subtilis HS3 and Bacillus mycoides EC18 are two rhizosphere-associated bacteria with plant growth-promoting activity. The CRISPR-Cas9 system was implemented to study various aspects of plant-microbe interaction mechanisms of these two environmental isolates. The results show that fengycin and surfactin are involved in the antifungal activity of B. subtilis HS3. Moreover, this strain emits several other volatile organic compounds (VOCs) than 2,3-butanediol, contributing to plant growth promotion. CLSM (Confocal laser scanning microscopy) observations of the GFP-labelled strain showed that HS3 selectively colonizes root hairs of grass (Lolium perenne) in a hydroponic system...
July 26, 2018: Environmental Microbiology
Kai Cai, Jun Yin, Hongmin Chao, Yaping Ren, Liping Jin, Yangrong Cao, Deqiang Duanmu, Zhongming Zhang
During the establishment of rhizobia-legume symbiosis, the cytokinin receptor LHK1 (Lotus Histidine Kinase 1) is essential for nodule formation. However, the mechanism by which cytokinin signaling regulates symbiosis remains largely unknown. In this study, an LHK1-interacting protein, LjCZF1, was identified and further characterized. LjCZF1 is a C3HC4-type RING finger protein that is highly conserved in plants. LjCZF1 specifically interacted with LHK1 in yeast two-hybrid, in vitro pull-down and co-immunoprecipitation assays conducted in tobacco...
July 26, 2018: Journal of Integrative Plant Biology
Douglas A Higgins, Megan K Young, Mary Tremaine, Maria Sardi, Jenna M Fletcher, Margaret Agnew, Lisa Liu, Quinn Dickinson, David Peris, Russell L Wrobel, Chris Todd Hittinger, Audrey P Gasch, Steven W Singer, Blake A Simmons, Robert Landick, Michael P Thelen, Trey K Sato
Imidazolium ionic liquids (IILs) have a range of biotechnological applications, including as pretreatment solvents that extract cellulose from plant biomass for microbial fermentation into sustainable bioenergy. However, residual levels of IILs, such as 1-ethyl-3-methyl imidazolium chloride ([C2 C1 im]Cl), are toxic to biofuel-producing microbes, including the yeast Saccharomyces cerevisiae. S. cerevisiae strains isolated from diverse ecological niches differ in genomic sequence and in phenotypes potentially beneficial for industrial applications, including tolerance to inhibitory compounds present in hydrolyzed plant feedstocks...
July 25, 2018: Genetics
William A Clayton, Nick W Albert, Amali H Thrimawithana, Tony K McGhie, Simon C Deroles, Kathy E Schwinn, Ben A Warren, Andrew R G McLachlan, Bowman L John, Brian R Jordan, Kevin M Davies
Damaging UVB radiation is a major abiotic stress facing land plants. In angiosperms the UV RESISTANCE LOCUS8 (UVR8) photoreceptor coordinates UVB responses, including inducing biosynthesis of protective flavonoids. We characterised the UVB responses of Marchantia polymorpha (marchantia), the model species for the liverwort group of basal plants. Physiological, chemical, and transcriptomic analyses were conducted on wild-type marchantia exposed to three different UVB regimes. CRISPR/Cas9 was used to obtain plant lines with mutations for components of the UVB signal pathway or the flavonoid biosynthetic pathway, and transgenics over-expressing the marchantia UVR8 sequence were generated...
July 25, 2018: Plant Journal: for Cell and Molecular Biology
Ayako Nishizawa-Yokoi, Nobutoshi Yamaguchi
To control the expression of their target genes, plant transcription factors bind to specific DNA sequences (called cis-elements) adjacent to the genes they regulate, thereby promoting or blocking the recruitment of RNA polymerase. Functional analysis of cis-elements is therefore essential for understanding transcriptional regulation, which underlies developmental programs and environmental responses. Using transgenic promoters containing mutations in their cis-elements, the roles of cis-elements in both transcriptional activity and transcription factor binding can be analyzed...
2018: Methods in Molecular Biology
Shigeo S Sugano, Ryuichi Nishihama
Plant transcription factors (TFs) belong to a wide variety of gene families. The systematic and rapid establishment of knockout lines of TF genes is critical for functional genetics. Genome engineering techniques for dissecting out the molecular function of TFs have been dramatically improved by CRISPR/Cas9-based genome editing technology. In the CRISPR/Cas9 system, Cas9 functions as a Cas9-gRNA ribonucleoprotein complex and uses its DNA endonuclease activity to induce the cleavage of the genome, which is targeted by gRNA...
2018: Methods in Molecular Biology
Ze Hong Lee, Nobutoshi Yamaguchi, Toshiro Ito
Gene editing enables the modification of genetic information in a targeted manner. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is a gene editing approach that has been widely used in both the animal and plant studies. Plant transcription factors play critical roles in plant development. Recent studies have used the CRISPER/Cas9 system to disrupt transcription factors and cis-elements in plants. In this chapter, we describe a protocol of using the CRISPR/Cas9 system to generate transcription factor mutants in the model plant species Arabidopsis thaliana...
2018: Methods in Molecular Biology
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