scholarly journals A simple Agrobacterium-mediated stable transformation technique for the hornwort model Anthoceros agrestis

2021 ◽  
Author(s):  
Eftychios Frangedakis ◽  
Manuel Waller ◽  
Tomoaki Nishiyama ◽  
Hirokazu Tsukaya ◽  
Xia Xu ◽  
...  

We have developed a simple Agrobacterium-mediated method for the stable transformation of the hornwort Anthoceros agrestis, the fifth bryophyte species for which a genetic manipulation technique becomes available. High transformation efficiency was achieved by using thallus tissue grown under low-light conditions. We generated a total of 216 transgenic A. agrestis lines expressing the β-Glucuronidase (GUS), cyan, green, and yellow fluorescent proteins under the control of the CaMV 35S promoter and several endogenous promoters. Nuclear and plasma membrane localization with multiple color fluorescent proteins was also confirmed. The transformation technique described here should pave the way for detailed molecular and genetic studies of hornwort biology, providing much needed insight into the molecular mechanisms underlying symbiosis, carbon-concentrating mechanism, RNA editing, and land plant evolution in general.

2019 ◽  
Vol 61 (2) ◽  
pp. 265-275 ◽  
Author(s):  
Hidenori Matsui ◽  
Hidekazu Iwakawa ◽  
Gang-Su Hyon ◽  
Izumi Yotsui ◽  
Shinpei Katou ◽  
...  

Abstract The evolution of adaptive interactions with beneficial, neutral and detrimental microbes was one of the key features enabling plant terrestrialization. Extensive studies have revealed conserved and unique molecular mechanisms underlying plant–microbe interactions across different plant species; however, most insights gleaned to date have been limited to seed plants. The liverwort Marchantia polymorpha, a descendant of early diverging land plants, is gaining in popularity as an advantageous model system to understand land plant evolution. However, studying evolutionary molecular plant–microbe interactions in this model is hampered by the small number of pathogens known to infect M. polymorpha. Here, we describe four pathogenic fungal strains, Irpex lacteus Marchantia-infectious (MI)1, Phaeophlebiopsis peniophoroides MI2, Bjerkandera adusta MI3 and B. adusta MI4, isolated from diseased M. polymorpha. We demonstrate that salicylic acid (SA) treatment of M. polymorpha promotes infection of the I. lacteus MI1 that is likely to adopt a necrotrophic lifestyle, while this effect is suppressed by co-treatment with the bioactive jasmonate in M. polymorpha, dinor-cis-12-oxo-phytodienoic acid (dn-OPDA), suggesting that antagonistic interactions between SA and oxylipin pathways during plant–fungus interactions are ancient and were established already in liverworts.


Author(s):  
Anita Roth-Nebelsick ◽  
Tatiana Miranda ◽  
Martin Ebner ◽  
Wilfried Konrad ◽  
Christopher Traiser

AbstractTrees are the fundamental element of forest ecosystems, made possible by their mechanical qualities and their highly sophisticated conductive tissues. The evolution of trees, and thereby the evolution of forests, were ecologically transformative and affected climate and biogeochemical cycles fundamentally. Trees also offer a substantial amount of ecological niches for other organisms, such as epiphytes, creating a vast amount of habitats. During land plant evolution, a variety of different tree constructions evolved and their constructional principles are a subject of ongoing research. Understanding the “natural construction” of trees benefits strongly from methods and approaches from physics and engineering. Plant water transport is a good example for the ongoing demand for interdisciplinary efforts to unravel form-function relationships on vastly differing scales. Identification of the unique mechanism of water long-distance transport requires a solid basis of interfacial physics and thermodynamics. Studying tree functions by using theoretical approaches is, however, not a one-sided affair: The complex interrelationships between traits, functionality, trade-offs and phylogeny inspire engineers, physicists and architects until today.


Science ◽  
2021 ◽  
Vol 371 (6531) ◽  
pp. eaba6605 ◽  
Author(s):  
Pierre-Marc Delaux ◽  
Sebastian Schornack

During 450 million years of diversification on land, plants and microbes have evolved together. This is reflected in today’s continuum of associations, ranging from parasitism to mutualism. Through phylogenetics, cell biology, and reverse genetics extending beyond flowering plants into bryophytes, scientists have started to unravel the genetic basis and evolutionary trajectories of plant-microbe associations. Protection against pathogens and support of beneficial, symbiotic, microorganisms are sustained by a blend of conserved and clade-specific plant mechanisms evolving at different speeds. We propose that symbiosis consistently emerges from the co-option of protection mechanisms and general cell biology principles. Exploring and harnessing the diversity of molecular mechanisms used in nonflowering plant-microbe interactions may extend the possibilities for engineering symbiosis-competent and pathogen-resilient crops.


2021 ◽  
Author(s):  
Eftychios Frangedakis ◽  
Manuel Waller ◽  
Tomoaki Nishiyama ◽  
Hirokazu Tsukaya ◽  
Xia Xu ◽  
...  

2013 ◽  
Vol 2013 ◽  
pp. 1-14 ◽  
Author(s):  
Maria S. Krasnikova ◽  
Denis V. Goryunov ◽  
Alexey V. Troitsky ◽  
Andrey G. Solovyev ◽  
Lydmila V. Ozerova ◽  
...  

PCR-based approach was used as a phylogenetic profiling tool to probe genomic DNA samples from representatives of evolutionary distant moss taxa, namely, classes Bryopsida, Tetraphidopsida, Polytrichopsida, Andreaeopsida, and Sphagnopsida. We found relatives of allPhyscomitrella patensmiR390 and TAS3-like loci in these plant taxa excluding Sphagnopsida. Importantly, cloning and sequencing ofMarchantia polymorphagenomic DNA showed miR390 and TAS3-like sequences which were also found among genomic reads ofM. polymorphaat NCBI database. Our data suggest that the ancient plant miR390-dependent TAS molecular machinery firstly evolved to target AP2-like mRNAs in Marchantiophyta and only then both ARF- and AP2-specific mRNAs in mosses. The presented analysis shows that moss TAS3 families may undergone losses of tasiAP2 sites during evolution toward ferns and seed plants. These data confirm that miR390-guided genes coding for ARF- and AP2-specific ta-siRNAs have been gradually changed during land plant evolution.


mBio ◽  
2018 ◽  
Vol 9 (5) ◽  
Author(s):  
Travis J. Wiles ◽  
Elena S. Wall ◽  
Brandon H. Schlomann ◽  
Edouard A. Hay ◽  
Raghuveer Parthasarathy ◽  
...  

ABSTRACTCorrelating the presence of bacteria and the genes they carry with aspects of plant and animal biology is rapidly outpacing the functional characterization of naturally occurring symbioses. A major barrier to mechanistic studies is the lack of tools for the efficient genetic manipulation of wild and diverse bacterial isolates. To address the need for improved molecular tools, we used a collection of proteobacterial isolates native to the zebrafish intestinal microbiota as a testbed to construct a series of modernized vectors that expedite genetic knock-in and knockout procedures across lineages. The innovations that we introduce enhance the flexibility of conventional genetic techniques, making it easier to manipulate many different bacterial isolates with a single set of tools. We developed alternative strategies for domestication-free conjugation, designed plasmids with customizable features, and streamlined allelic exchange using visual markers of homologous recombination. We demonstrate the potential of these tools through a comparative study of bacterial behavior within the zebrafish intestine. Live imaging of fluorescently tagged isolates revealed a spectrum of distinct population structures that differ in their biogeography and dominant growth mode (i.e., planktonic versus aggregated). Most striking, we observed divergent genotype-phenotype relationships: several isolates that are predicted by genomic analysis andin vitroassays to be capable of flagellar motility do not display this trait within living hosts. Together, the tools generated in this work provide a new resource for the functional characterization of wild and diverse bacterial lineages that will help speed the research pipeline from sequencing-based correlations to mechanistic underpinnings.IMPORTANCEA great challenge in microbiota research is the immense diversity of symbiotic bacteria with the capacity to impact the lives of plants and animals. Moving beyond correlative DNA sequencing-based studies to define the cellular and molecular mechanisms by which symbiotic bacteria influence the biology of their hosts is stalling because genetic manipulation of new and uncharacterized bacterial isolates remains slow and difficult with current genetic tools. Moreover, developing tools de novo is an arduous and time-consuming task and thus represents a significant barrier to progress. To address this problem, we developed a suite of engineering vectors that streamline conventional genetic techniques by improving postconjugation counterselection, modularity, and allelic exchange. Our modernized tools and step-by-step protocols will empower researchers to investigate the inner workings of both established and newly emerging models of bacterial symbiosis.


2017 ◽  
Vol 372 (1730) ◽  
pp. 20160380 ◽  
Author(s):  
Marcel Dann ◽  
Dario Leister

Although some elements of the photosynthetic light reactions might appear to be ideal, the overall efficiency of light conversion to biomass has not been optimized during evolution. Because crop plants are depleted of genetic diversity for photosynthesis, efforts to enhance its efficiency with respect to light conversion to yield must generate new variation. In principle, three sources of natural variation are available: (i) rare diversity within extant higher plant species, (ii) photosynthetic variants from algae, and (iii) reconstruction of no longer extant types of plant photosynthesis. Here, we argue for a novel approach that outsources crop photosynthesis to a cyanobacterium that is amenable to adaptive evolution. This system offers numerous advantages, including a short generation time, virtually unlimited population sizes and high mutation rates, together with a versatile toolbox for genetic manipulation. On such a synthetic bacterial platform, 10 000 years of (crop) plant evolution can be recapitulated within weeks. Limitations of this system arise from its unicellular nature, which cannot reproduce all aspects of crop photosynthesis. But successful establishment of such a bacterial host for crop photosynthesis promises not only to enhance the performance of eukaryotic photosynthesis but will also reveal novel facets of the molecular basis of photosynthetic flexibility. This article is part of the themed issue ‘Enhancing photosynthesis in crop plants: targets for improvement’.


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