scholarly journals CRISPR-Cas12a Genome Editing at the Whole-Plant Level Using Two Compatible RNA Virus Vectors

2021 ◽  
Author(s):  
Mireia Uranga ◽  
Marta Vazquez-Vilar ◽  
Diego Orzáez ◽  
José-Antonio Daròs
2021 ◽  
Author(s):  
Mireia Uranga ◽  
Marta Vazquez-Vilar ◽  
Diego Orzaez ◽  
Jose-Antonio Daros

The use of viral vectors that can replicate and move systemically through the host plant to deliver bacterial clustered, regularly interspaced, short palindromic repeats (CRISPR) components enables genome editing at the whole-plant level and avoids the requirement for labor-intensive stable transformation. However, this approach usually relies on previously transformed plants that stably express a CRISPR-associated (Cas) nuclease. Here we describe successful DNA-free genome editing of Nicotiana benthamiana using two compatible RNA virus vectors, derived from tobacco etch virus (TEV; genus Potyvirus) and potato virus X (PVX; genus Potexvirus), which replicate in the same cells. The TEV and PVX vectors respectively express a Cas12a nuclease and the corresponding guide RNA. This novel two-virus vector system improves the toolbox for transformation-free virus-induced genome editing in plants and will advance efforts to breed more nutritious, resistant, and productive crops.


2021 ◽  
Author(s):  
Tingdong Li ◽  
Jiacheng Hu ◽  
Yu Sun ◽  
Boshu Li ◽  
Dingliang Zhang ◽  
...  

1997 ◽  
Vol 11 (2) ◽  
pp. 241-247 ◽  
Author(s):  
Christy L. Sprague ◽  
Edward W. Stoller ◽  
Loyd M. Wax

Five biotypes of common cocklebur that were not controlled with acetolactate synthase (ALS)-inhibiting herbicides were tested in greenhouse and laboratory studies to determine the magnitude of resistance and cross-resistance to four ALS-inhibiting herbicides. In vivo inhibition of ALS was also evaluated. Based on phytotoxicity, all five ALS-resistant biotypes of common cocklebur were > 390 times more resistant than the susceptible biotype to imazethapyr. However, only four of these biotypes were also resistant to another imidazolinone, imazaquin. Two biotypes were cross-resistant to the sulfonylurea, chlorimuron, and the triazolopyrimidine sulfonanilide, NAF-75. One biotype demonstrated intermediate susceptibility to imazaquin, chlorimuron, and NAF-75. In all cases, the resistance exhibited at the whole plant level was associated with an insensitive ALS.


2013 ◽  
Vol 765-767 ◽  
pp. 2971-2975 ◽  
Author(s):  
Xue Gong ◽  
Ming Li Liu ◽  
Li Jun Zhang ◽  
Wei Liu ◽  
Che Wang

Sucrose transporters (SUCs or SUTs) are considered as the important carriers and responsible for the loading, unloading and distribution of sucrose, but at present there is no report that SUCs are involved in sucrose distribution and metabolism under drought stress at the whole-plant level. AtSUC4, as the unique member of SUT4-clade inArabidopsis thaliana, may be important for plant stress tolerance. Here, by analyzing two homozygous mutation lines ofAtSUC4(Atsuc4-1andAtsuc4-2), we found drought stress induced higher sucrose, lower fructose and glucose contents in shoots, and lower sucrose, higher fructose and glucose contents in roots of these mutants compared with the wild-type (WT), leading to an imbalance of sucrose distribution, fructose and glucose (sucrose metabolites) accumulation changes at the whole-plant level. Thus we believe thatAtSUC4regulates sucrose distribution and metabolism in response to drought stress.


2018 ◽  
Vol 36 (3) ◽  
pp. 330-340 ◽  
Author(s):  
Adalberto Di Benedetto ◽  
Claudio Galmarini ◽  
Jorge Tognetti

ABSTRACT Climbing Epipremnum aureum plants develop larger leaves than unsupported, hanging plants. This effect may be regarded, in part, as a thigmomorphogenic response, but gravimorphogenetic effect may also be involved, since polar auxin transport is known to be negatively affected in plants with horizontal or hanging stems, which may result in an altered hormone balance at the whole plant level. The present work was aimed at studying how exogenous auxins and cytokinins may influence growth of E. aureum rooted cuttings under different training systems. Rooted cuttings of E. aureum were cultivated either climbing on an upright wooden board or creeping on the glasshouse bench or hanging from a basket. All leaves of each plant were sprayed to run-off at sunset with four indole-3-acetic acid (IAA) doses 7 days after transplanting and one week later, with four benzylaminopurine (BAP) concentrations, rendering 16 hormone combination treatments. The application of IAA or BAP at 50 mg L-1 to creeping and hanging plants significantly promoted growth but, in climbing plants, a negative effect was generally observed. Changes in net assimilation and photosynthetic rates, together with modified allometric coefficients, accounted for these responses. The higher growth promotion by exogenous growth regulators observed in creeping or hanging plants compared to climbing plants, may be interpreted mostly as a gravimorphogenetic response.


Author(s):  
Manutsawan Manokieng ◽  
◽  
Arunothai Jampeetong ◽  

Abstract The effects of supplemental cations on growth, nitrogen, and mineral accumulation were assessed in Canna indica L. Similar sized 45 days-old plants were grown on a nutrient solution modified from Hoagland and Arnon (1950). The different cations were added to generate 6 treatments (n=4): (i) control (no cation added), (ii) 2.5 mM K+, (iii) 2.5 mM Ca2+, (iv) 75 mM Na+, (v) 1.25 mM K+ + 1.25 mM Ca2+ and (vi) 2.5 mM Ca2+ + 75 mM Na+, respectively. An experiment was carried out in the greenhouse for 49 days. The study found that supplemental K+ and K++ Ca2+ increased plant growth and total biomass. The highest SER was found in plants receiving supplemental K+. In contrast, SERs, leaf areas, and total biomass decreased in Na+ or Na++Ca2+ supplemented plants. The accumulated NO3- concentration (at the whole plant level) was also highest in the plants with supplemental K+ and K++Ca2+. The total nitrogen accumulation was higher in the K+, Ca2+, and K++Ca2+ supplemented plants than in the control plants. The results suggest that supplemental cations particularly K+ can enhance plant growth and nitrogen accumulation in C. indica. Therefore, cation supplementation could be an alternative technique to stimulate plant growth and improve nitrate removal in constructed wetlands. Keywords: Constructed wetland, Nitrate removal, Potassium, Tropical wetland plants


2021 ◽  
Vol 12 ◽  
Author(s):  
Laura Siles ◽  
Kirsty L. Hassall ◽  
Cristina Sanchis Gritsch ◽  
Peter J. Eastmond ◽  
Smita Kurup

Seed yield is a complex trait for many crop species including oilseed rape (OSR) (Brassica napus), the second most important oilseed crop worldwide. Studies have focused on the contribution of distinct factors in seed yield such as environmental cues, agronomical practices, growth conditions, or specific phenotypic traits at the whole plant level, such as number of pods in a plant. However, how female reproductive traits contribute to whole plant level traits, and hence to seed yield, has been largely ignored. Here, we describe the combined contribution of 33 phenotypic traits within a B. napus diversity set population and their trade-offs at the whole plant and organ level, along with their interaction with plant level traits. Our results revealed that both Winter OSR (WOSR) and Spring OSR (SOSR); the two more economically important OSR groups in terms of oil production; share a common dominant reproductive strategy for seed yield. In this strategy, the main inflorescence is the principal source of seed yield, producing a good number of ovules, a large number of long pods with a concomitantly high number of seeds per pod. Moreover, we observed that WOSR opted for additional reproductive strategies than SOSR, presenting more plasticity to maximise seed yield. Overall, we conclude that OSR adopts a key strategy to ensure maximal seed yield and propose an ideal ideotype highlighting crucial phenotypic traits that could be potential targets for breeding.


2016 ◽  
Author(s):  
Χρυσάνθη Καλλονιάτη

Symbiotic nitrogen fixation in legumes takes place in specialized organs called nodules,which become the main source of assimilated nitrogen for the whole plant. Symbiotic nitro‐gen fixation requires exquisite integration of plant and bacterial metabolism and involvesglobal changes in gene expression and metabolite accumulation in both rhizobia and thehost plant. In order to study the metabolic changes mediated by symbiotic nitrogen fixationon a whole‐plant level, metabolite levels were profiled by gas chromatography–mass spec‐trometry in nodules and non‐symbiotic organs of Lotus japonicus plants uninoculated or in‐oculated with M. loti wt,  ΔnifA or  ΔnifH fix‐ strains. Furthermore, transcriptomic andbiochemical approaches were combined to study sulfur metabolism in nodules, its link tosymbiotic nitrogen fixation, and the effect of nodules on whole‐plant sulfur partitioning andmetabolism. It is well established that nitrogen and sulfur (S) metabolism are tightly en‐twined and sulfur is required for symbiotic nitrogen fixation, however, little is known aboutthe molecular and biochemical mechanisms governing sulfur uptake and assimilation duringsymbiotic nitrogen fixation. Transcript profiling in Lotus japonicus was combined with quan‐tification of S‐metabolite contents and APR activity in nodules and in non‐symbiotic organsof plants uninoculated or inoculated with M. loti wt, ΔnifA or ΔnifH fix‐ strains. Moreover,sulfate uptake and its distribution into different plant organs were analyzed and 35S‐flux intodifferent S‐pools was monitored. Metabolite profiling revealed that symbiotic nitrogen fixa‐tion results in dramatic changes of many aspects of primary and secondary metabolism innodules which leads to global reprogramming of metabolism of the model legume on awhole‐plant level. Moreover, our data revealed that nitrogen fixing nodules represent athiol‐rich organ. Their high APR activity and 35S‐flux into cysteine and its metabolites in com‐bination with the transcriptional up‐regulation of several genes involved in sulfur assimila‐tion highlight the function of nodules as a new site of sulfur assimilation. The higher thiolcontent observed in non‐symbiotic organs of nitrogen fixing plants in comparison touninoculated plants cannot be attributed to local biosynthesis, indicating that nodules couldserve as a novel source of reduced sulfur for the plant, which triggers whole‐plant repro‐gramming of sulfur metabolism. Interestingly, the changes in metabolite profiling and theenhanced thiol biosynthesis in nodules and their impact on the whole‐plant sulfur, carbonand nitrogen economy are dampened in fix‐ plants, which in most respects metabolically re‐sembled uninoculated plants, indicating a strong interaction between nitrogen fixation andsulfur and carbon metabolism.


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