Abstract：
:Accurate and timely detection of transgene copy number in sugarcane is currently hampered by the requirement to use Southern blotting, needing relatively large amounts of genomic DNA and, therefore, the continued growth and maintenance of bulky plants in containment glasshouses. In addition, the sugarcane genome is both polyploid and aneuploid, complicating the identification of appropriate genes for use as references in the development of a high-throughput method. Using bioinformatic techniques followed by in vitro testing, two genes that appear to occur once per base genome of sugarcane were identified. Using these genes as reference genes, a high-throughput assay employing RT-qPCR was developed and tested using a group of sugarcane plants that contained unknown numbers of copies of the nptII gene encoding kanamycin resistance. Using this assay, transgene copy numbers from 3 to more than 50 were identified. In comparison, Southern blotting accurately identified the number of transgene copies for one line and by inference for another, but was not able to provide an accurate estimation for transgenic lines containing numerous copies of the nptII gene. Using the reference genes identified in this study, a high-throughput assay for the determination of transgene copy number was developed and tested for sugarcane. This method requires much less input DNA, can be performed much earlier in the production of transgenic sugarcane plants and allows much more efficient assessment of numerous potentially transgenic lines than Southern blotting.
journal_name
Plant Cell Repjournal_title
Plant cell reportsauthors
Casu RE,Selivanova A,Perroux JMdoi
10.1007/s00299-011-1150-7subject
Has Abstractpub_date
2012-01-01 00:00:00pages
167-77issue
1eissn
0721-7714issn
1432-203Xjournal_volume
31pub_type
杂志文章abstract：:The DNA-binding domain of the yeast transcriptional activator GAL4 was expressed in transgenic tobacco plants in order to attempt to obtain specific repression of reporter genes. Expression of the GUS and the NPTII gene was essentially the same regardless of the presence of a gene encoding a functional GAL4 DNA-bindin...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/BF00232920
更新日期：1995-10-01 00:00:00
abstract：:Growth inhibition towards Rhizopus nigricans, Fusarium oxysporum f. sp. radicis-lycopersici, Verticillium albo-atrum and Pythium ultimum was observed in vitro using a purified chitosanase from an actinomycete, Streptomyces sp, strain N174. The corresponding gene, with its own signal peptide, was inserted into pBI121.7...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/BF00193724
更新日期：1995-12-01 00:00:00
abstract：:A novel member of the AP2/ERF transcription factor family, SlERF5, was identified from a tomato mature leaf cDNA library screen. The complete DNA sequence of SlERF5 encodes a putative 244-amino acid DNA-binding protein which most likely acts as a transcriptional regulator and is a member of the ethylene responsive fac...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-011-1170-3
更新日期：2012-02-01 00:00:00
abstract：:The ability to germinate orchids from seeds in vitro presents a useful and viable method for the propagation of valuable germplasm, maintaining the genetic heterogeneity inherent in seeds. Given the ornamental and medicinal importance of many species within the genus Dendrobium, this review explores in vitro technique...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-015-1829-2
更新日期：2015-10-01 00:00:00
abstract：:Brassica juncea L. is a Zn/Cd accumulator. To determine the physiological basis of its metal accumulation phenotype, the functional properties and role of the metal efflux transporter BjCET2 were investigated using transgenic technology. Heterologous expression of BjCET2 in the double mutant yeast strain Deltazrc1Delt...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-009-0723-1
更新日期：2009-08-01 00:00:00
abstract：:A method for regenerating pigeonpea [Cajanus cajan (L.) Millsp.] plants has been developed using distal cotyledonary segments of mature seeds as explants. A large number of shoot buds were induced directly from explants of genotypes T-15-15 and GAUT-82-90 when cultured on six different basal media fortified with 22.2 ...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s002990050469
更新日期：1998-06-01 00:00:00
abstract：:Plating rice anthers on a semisolid induction medium containing 250 or 500 mg/l colchicine for 24 or 48 h-incubations followed by transfer to colchicine-free medium and standard anther culture procedures resulted in overall 1.5- to 2.5- fold increases in doubled haploid green plant productions compared to control anth...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/BF00231961
更新日期：1994-05-01 00:00:00
abstract：: Salvia miltiorrhiza contains two groups of biologically active secondary metabolites termed phenolic compounds (e.g. rosmarinic acid) and tanshinones (e.g. cryptotanshinone). Their roles in plant defense responses were examined using a simplified system consisting of a yeast elicitor and a Ti C58 transformed S. milti...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s002999900166
更新日期：2000-06-01 00:00:00
abstract：KEY MESSAGE:We demonstrate the role of DREB1A transcription factor in better root and shoot partitioning and higher transpiration efficiency in transgenic chickpea under drought stress Chickpea (Cicer arietinum L.) is mostly exposed to terminal drought stress which adversely influences its yield. Development of cultiva...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-014-1699-z
更新日期：2015-02-01 00:00:00
abstract：:Reliable regeneration protocols for Eucalyptus camaldulensis using leaf explants from in vitro-grown plants have been developed. Out of the 24 clones tested 13 were regenerated and of these, 6 showed regeneration from more than 60% of the explants. Identical protocols were also successful in the regeneration of some c...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s002990050321
更新日期：1997-09-01 00:00:00
abstract：:We developed procedures for slow-growth storage of Cedrus atlantica and Cedrus libani microcuttings of juvenile and adult origin, noting factors favouring the extension of subculture intervals. Microcuttings could be stored effectively up to 6 months at 4 degrees C and reduced light intensity, provided that they were ...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-006-0129-2
更新日期：2006-07-01 00:00:00
abstract：:A protocol for Agrobacterium-mediated transformation with mannose selection was developed for cotyledon petiole, hypocotyl and leaf explants of tomato (Lycopersicon esculentum L. Mill). More than 400 transgenic plants from three tomato varieties were selected with 1% mannose in combination with 0.1-0.5% glucose. Avera...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-004-0809-8
更新日期：2004-10-01 00:00:00
abstract：:A biolistic transformation procedure was used to transform embryogenic Pinus radiata tissue with constructs containing the Zea mays UBI1 (ubiquitin)-promoter followed by the P. radiata CAD (cinnamyl alcohol dehydrogenase) cDNA in sense or anti-sense orientation or in the form of an inverted-repeat. The effect of the d...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-004-0911-y
更新日期：2005-05-01 00:00:00
abstract：:Optimal protoplast yields from cotyledons (2.0×10(6) protoplasts/ 0.5 g tissue) and from true leaves (5.0×10(6) protoplasts/g tissue) of two Cucumis sativus genotypes were obtained following a 16 h digestion with, respectively, 1.25% pectinase+0.5% Cellulysin and 0.5 % pectinase+ 1.0% Cellulysin. Enzyme solutions were...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/BF00231549
更新日期：1990-07-01 00:00:00
abstract：KEY MESSAGE:Two lines of Taxus × media hairy roots harbouring or not the TXS transgene demonstrated diverse gene expression and taxane yield during cultivation in PFD-supported two liquid-phase culture system. Two lines of Taxus × media hairy roots were subjected to single or twice-repeated supplementation with methyl ...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-018-2351-0
更新日期：2019-01-01 00:00:00
abstract：:The use of antibiotic or herbicide resistant genes as selection markers for production of transgenic plants and their continuous presence in the final transgenics has been a serious problem for their public acceptance and commercialization. MAT (multi-auto-transformation) vector system has been one of the different st...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-010-0974-x
更新日期：2011-04-01 00:00:00
abstract：:Pyrolysis mass spectrometry (PyMS) is a rapid, simple, high-resolution analytical method based on thermal degradation of complex material in a vacuum and has been widely applied to the discrimination of closely related microbial strains. Leaf samples of six species and one variety of higher plants (Rosa multiflora, R....
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-003-0714-6
更新日期：2004-02-01 00:00:00
abstract：KEY MESSAGE:Sustainability and safety aspects of plant cell cultures as food are presented. Applicability of dairy side streams as carbon source and use of natural growth enhancers in cultivation are shown. Biotechnologically produced cellular products are currently emerging to replace and add into the portfolio of agr...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-020-02592-2
更新日期：2020-12-01 00:00:00
abstract：:In this paper we describe a simple method to improve the plating efficiency in plant cell cultures.Two-stage plating is used; in the first stage the cells are inoculated at high density in 0.2% agarized culture medium for ten days to facilitate growth; under this condition, each cell produces a single micro-colony tra...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/BF00571305
更新日期：1985-05-01 00:00:00
abstract：:Sweet cherry shows S-RNase-based gametophytic self-incompatibility, which prevents self- and cross-fertilization between genetically related individuals. The specificity of the self-incompatible reaction is determined by two genes located in the S-locus. These encode a pistil-expressed ribonuclease (S-RNase) that inhi...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-010-0857-1
更新日期：2010-07-01 00:00:00
abstract：:In the present paper, attempts were made to explore the possibility of employing ultraviolet (UV) irradiation in citrus asymmetric fusion for transfer of limited amount of favorable traits from a desirable cultivar to a target one. Exposure of Satsuma mandarin (Citrus unshiu Marc.) embryogenic protoplasts to UV at an ...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-007-0350-7
更新日期：2007-08-01 00:00:00
abstract：:Seeds are heterogeneous storage reserves with wide array of storage compounds that include various soluble carbohydrates, starch polymer, storage proteins and lipids. These stored reserves comprise 70% of the world's caloric intake in the form of food and animal feed produced through sustainable agriculture, which con...
journal_title：Plant cell reports
pub_type： 社论
doi：10.1007/s00299-017-2135-y
更新日期：2017-05-01 00:00:00
abstract：:Isolated ovules from the maize homozygous recessive brown midrib (bm3) were in vitro pollinated by pollen carrying the dominant allele - and the purple embryo marker (PEM). The colour characters of the embryos and kernels corresponded to the results of control pollination and confirmed the process of double fertilizat...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/BF00267652
更新日期：1981-08-01 00:00:00
abstract：KEY MESSAGE:Wheat MAPK gene TaMPK14 is N starvation response and is crucial in modulating plant low-N stress tolerance. Improving plant N use efficiency (NUE) contributes largely to the sustainable crop production worldwide. In this study, TaMPK14, a mitogen-activated protein kinase (MAPK) family gene in T. aestivum, w...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-020-02564-6
更新日期：2020-10-01 00:00:00
abstract：:We developed an efficient procedure for transformation and regeneration of L. esculentum cv. Moneymaker from cotyledon explants. The effect of two parameters on the transformation frequency was investigated in detail. The use of feeder layers during cocultivation proved to be critical. In addition, it was found that A...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/BF00232816
更新日期：1993-09-01 00:00:00
abstract：:Stem sections from shoot cultures maintained in vitro were used to produce transgenic plants of the potato, Solanum tuberosum L. cv. 'Russet Burbank'. Stem internode pieces inoculated with Agrobacterium tumefaciens containing coat protein genes from potato virus X and potato virus Y, produced shoots with a frequency o...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/BF00233028
更新日期：1991-05-01 00:00:00
abstract：:Quantitative real-time polymerase chain reaction (PCR) assays were designed that enabled the zygosity of transgenes in soybean [Glycine max (L.) Merrill] and peanut (Arachis hypogaea L.) to be determined. The two zygosity assays, based on TaqMan technology that uses a fluorogenic probe which hybridizes to a PCR target...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s002990100326
更新日期：2001-07-01 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：KEY MESSAGE:A novel rice constitutive promoter (P OsCon1 ) was isolated. The molecular mechanism of the promoter activity was investigated. P OsCon1 could be used as an alternative constitutive promoter for crop transgenic engineering. Monocot constitutive promoter is an important resource for crop transgenic engineeri...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s00299-014-1644-1
更新日期：2014-10-01 00:00:00
abstract：: A high frequency of embryogenesis and transformation from all parts of flowers of two lines of Medicago truncatula R-108-1 and Jemalong J5 were obtained. Using this flower system, we obtained transgenic plants expressing promoter-uidA gene fusions as well as the gfp living cell color reporter gene. Moreover, this met...
journal_title：Plant cell reports
pub_type： 杂志文章
doi：10.1007/s002999900168
更新日期：2000-06-01 00:00:00
