Abstract:
KEY MESSAGE:The recovery of non-functional-enhanced green fluorescence protein can be used as indicator to facilitate the identification of mutants generated by CRISPR/Cas9. The CRISPR/Cas9 system is a powerful tool for genome editing and it has been employed to knock out genes of interest in multiple plant species. Identification of desired mutants from regenerated plants is necessary prior to functional study. Current screening methods work based on the purification of genomic DNA and it would be laborious and time consuming using these methods to screen mutants from a large population of seedlings. Here, we developed the non-functional enhanced green fluorescence protein (nEGFP) reporter gene by inserting a single guide RNA (sgRNA) and the protospacer adjacent motif in the 5' coding region of EGFP, and the activity of nEGFP could be recovered after successful targeted editing. Using the nEGFP as the reporter gene in Nicotiana tabacum, we found that over 94% of the plants exhibiting EGFP fluorescence were confirmed to be desired mutants. The use of this nEGFP reporter construct had limited negative effect on editing efficiency, and the expression of Cas9 and sgRNA was not affected. Moreover, this method was also applied in grape by targeting the phytoene desaturase gene (PDS), and the grape cells with EGFP signal were revealed to contain targeted mutations in VvPDS. Our results show that the nEGFP gene can be used as reporter to help screen mutants according to the recovered EGFP fluorescence during the application of CRISPR/Cas9 in plants.
journal_name
Plant Cell Repjournal_title
Plant cell reportsauthors
Ren C,Guo Y,Gathunga EK,Duan W,Li S,Liang Zdoi
10.1007/s00299-019-02465-3subject
Has Abstractpub_date
2019-12-01 00:00:00pages
1541-1549issue
12eissn
0721-7714issn
1432-203Xpii
10.1007/s00299-019-02465-3journal_volume
38pub_type
杂志文章abstract::Yucca valida is an important potential source of steroidal saponins closely related to Yucca schidigera, the species that is commercially exploited from the wild as a source of steroidal extracts. Neither of the species has been domesticated mainly because of their slow growth and long life span before harvesting. Her...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-006-0144-3
更新日期:2006-08-01 00:00:00
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-014-1670-z
更新日期:2014-12-01 00:00:00
abstract::Shoot cultures of Allium sativum L. were established from stem-tip and scape-tip. Various combinations of media, growth regulators were compared. The best establishment and proliferation of stem-tip and scape-tip explants were obtained on MS medium containing 0.5 mg l(-1) NAA and 0.5 mg l(-1) KT. The roots were produc...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00233301
更新日期:1994-11-01 00:00:00
abstract:: Identifying transformed plant lines carrying the antibiotic resistance marker gene, neomycin phosphotransferase II, requires a more definitive test than the ability of the plant to grow on kanamycin. Although a number of alternative assays have been described, most are cumbersome, time consuming and/or require the us...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s002990050014
更新日期:2000-01-01 00:00:00
abstract::Microcallus (more than 60 cells) formation was obtained from leaf mesophyll protoplasts of 6 species and varieties in the genus Actinidia Lindl. (kiwifruit). The best results were achieved by using liquid over agarose culture for A. arguta var. arguta, liquid and agarose disc type culture for A. arguta var. purpurea, ...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00231583
更新日期:1996-09-01 00:00:00
abstract::Murashige and Skoog's modified medium in 1% Difco Bacto-agar supplemented with sugar alcohols (sorbsitol, mannitol), growth regulators (1-naphthalenacetic acid, 2,4-dichlorophenoxyacetic acid, benzyladenine, kinetin) and sugars (fructose, glucose, sucrose) induced aposporous gametophytes from pinnae of Pteris vittata ...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00232608
更新日期:1991-10-01 00:00:00
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF01146782
更新日期:1997-03-01 00:00:00
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-011-1080-4
更新日期:2011-09-01 00:00:00
abstract::The ample supply of O2 proved to be of great importance for berberine production in cell suspension culture ofThalictrum minus, as the specific O2 consumption rate of berberine-producing cells was twice as high as that of non-producing cells. Furthermore, berberine yield increased with increases in the volumetric O2 t...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00778546
更新日期:1989-04-01 00:00:00
abstract::A DNA fragment encoding a 12-amino acid (aa) HIV-1 Tat transduction peptide fused to a 90-aa murine rotavirus NSP4 enterotoxin protein (Tat-NSP4(90)) was transferred to Solanum tuberosum by Agrobacterium tumefaciens-mediated transformation. The fusion gene was detected in the genomic DNA of transformed plant leaf tiss...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-003-0697-3
更新日期:2004-01-01 00:00:00
abstract::Expression of the green fluorescent protein (gfp) gene, under regulatory control of either the constitutive 35S promoter or the developmentally-regulated lectin promoter was monitored and quantified using a newly-developed automated tracking system. The automated system consisted of a computer-controlled two-dimension...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-006-0142-5
更新日期:2006-09-01 00:00:00
abstract:KEY MESSAGE:SnYSL3 encodes a plasma-localized transporter delivering various metal-nicotianamine complexes. The expression of SnYSL3 is up-regulated by excess Cd, suggesting an important role for SnYSL3 in response to Cd stress. The Yellow Stripe-Like (YSL) transporters have been proposed to participate in metal uptake...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-016-2079-7
更新日期:2017-02-01 00:00:00
abstract::Axillary buds from 5 genotypes of mulberry belonging to 4 species were cultured on modified MS basal medium. A total of 30 media combinations were tried for all the genotypes. The response of axillary buds and the requirement for growth regulators varied with genotype. In Morus indica BAP (0.25-0.5 mg/l), and in M. al...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00272107
更新日期:1990-05-01 00:00:00
abstract::Leaf blade tissue of maize inbred lines B73 and Mo17 was analyzed for intraspecific genetic variability in the heat shock response. The maize inbreds were characterized for acquired thermal tolerance and patterns of heat shock protein synthesis. The leakage conductivity assay of membrane stability during stress indica...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00233096
更新日期:1992-10-01 00:00:00
abstract::To study the developmental transition of chloroplasts from C(3) to C(4) photosynthesis in the terrestrial single-cell C(4) species Bienertia sinuspersici, a regeneration protocol was developed. Stem explant material developed callus either with or without red nodular structures (RNS) when cultured on Murashige-Skoog (...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-011-1067-1
更新日期:2011-08-01 00:00:00
abstract::Protoplasts isolated from cell suspensions of albinoMedicago borealis andM. sativa were fused chemically, using two methods, and electrically. Although a small scale method of chemical fusion gave the highest fusion frequency, electrofusion was the superior technique on the basis of throughput of green somatic hybrid ...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00735772
更新日期:1989-05-01 00:00:00
abstract::The present protocol was aimed at establishing a routine transformation procedure via Agrobacterium tumefaciens for an important Oncidium orchid cultivar. An expression vector containing hptII and gusA genes driven by the cauliflower mosaic virus (CaMV) 35S promoter was successfully introduced into the Oncidium orchid...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-003-0614-9
更新日期:2003-06-01 00:00:00
abstract::Processes involved in transformation of regenerable soybean (Glycine max (L.) Merrill) immature zygotic cotyledons were studied by assaying the transient expression of the β-glucuronidase gene driven by the 35S promoter and terminated at the 3' end by the soybean 7S storage protein gene. The plasmid containing the chi...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00234510
更新日期:1994-07-01 00:00:00
abstract::Three different regeneration systems, viz. direct regeneration of adventitious shoot buds from explant, regeneration through callus cultures and somatic embryos were compared to see their effect on transfer of neomycin phosphotransferase (nptII) and beta-glucuronidase (GUS) reporter gene (gus) to Morus alba clone M5, ...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-006-0217-3
更新日期:2007-02-01 00:00:00
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00232455
更新日期:1996-04-01 00:00:00
abstract::In Materials and method section, a sweetpotato variety "Taizhong-6" (China national number 2013003) should be renamed as Ayamurasaki". ...
journal_title:Plant cell reports
pub_type: 杂志文章,已发布勘误
doi:10.1007/s00299-019-02487-x
更新日期:2020-01-01 00:00:00
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-014-1725-1
更新日期:2015-03-01 00:00:00
abstract::The aim of this study was the improvement of embryo production in wheat anther culture. Three butanol alcohols, n-butanol, sec-butanol and tert-butanol, were evaluated for their effect on microspore embryogenesis in two spring cultivars of wheat, Pavon and Caramba. Application of n-butanol, at 0.1 and 0.2% (v/v) in th...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-007-0500-y
更新日期:2008-05-01 00:00:00
abstract::Rice plants (Oryza sativa L., Chinsurah Boro II var. Indica) were regenerated from protoplasts isolated from microspore derived cell suspensions. A simple procedure for the establishment of such cell suspension cultures from embryogenic microcallus derived from cultured isolated microspores of Indica-type rice is desc...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00232295
更新日期:1990-09-01 00:00:00
abstract::A series of RAPD markers generated by a single 10-mer primer were analyzed by hybridization to amplified and genomic DNA and by sequencing in two Brassica species. Primer B18 produced different profiles of nine major bands each in both Brassica nigra (B genome) and B. napus (AC genomes). Cloning and sequencing of five...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00232727
更新日期:1995-07-01 00:00:00
abstract:KEY MESSAGE:Two translation-related proteins are identified as FMT-interacting proteins. However, FMT, unlike mutants of other CLU genes in fly and human, has no clear impact on the accumulation of mitochondrial proteins. Organelle distribution is critical for effective metabolism and stress response and is controlled ...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-020-02634-9
更新日期:2021-02-01 00:00:00
abstract::Scanning electron microscopy has been used to investigate the morphological changes occurring during the development of alfalfa somatic embryos. Embryos were initiated from callus, transferred to suspension culture and matured on solid agar medium. This developmental pattern was compared to that of zygotic embryos dev...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00235082
更新日期:1992-06-01 00:00:00
abstract::An in vitro propagation technique based on axillary bud proliferation has been developed for matureSapium sebiferum trees. Nodal segments cultured on Murashige and Skoog (MS) medium supplemented with benzyl adenine (1-10 μM and α-naphthaleneacetic acid (0-0.5 μM showed axillary bud proliferation. Shoots proliferated i...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF01275506
更新日期:1997-06-01 00:00:00
abstract::Low initiation frequency is one of the main barriers in applying somatic embryogenesis to the clonal production of Pinus species. Factors affecting initiation, including basal medium, plant growth regulators, and Phytagel concentration, have been investigated in loblolly pine (Pinus taeda L.). BM1 basal medium proved ...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s002990050396
更新日期:1998-02-01 00:00:00
abstract::In vitro adventitious shoot formation was examined in petioles and laminae ofPaulownia elongata, P. fortunei, andP. 'Henan 1'. Fully expanded, dark green, thick, older leaves were healthier and exhibited greater callus and shoot production than did young leaves. The ten clones used varied greatly in adventitious shoot...
journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF01088288
更新日期:1997-02-01 00:00:00