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Leaf Disk Transformation Using Agrobacterium tumefaciens-Expression of Heterologous Genes in Tobacco
Time of Update: 2021-01-20
The protocol presented here works well in our hands withNicotiana tabacum cultivar “petit havana” mutant SR1 ( 2 ) andAgrobacterium tumefaciens strain LBA4404 ( 3 ) harboring binary vectors conferring kanamycin resistance (100 mg/L).
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Production of Hairy Root Cultures and Transgenic Plants by Agrobacterium rhizogenes-Mediated Transformation
Time of Update: 2021-01-20
Agrobacterium rhizogenes -mediated transformation results in the development of hairy roots at the site of infection.
rhizogenes and the subsequent selection of hairy roots on hormone-free medium.
In addition, transgenic plants regenerated from hairy roots often show an altered phenotype due to the presence of therol genes.
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Proteomics and the Analysis of Nodulation
Time of Update: 2021-01-20
Application of proteomics is becoming increasingly important to understand the function of genes and their encoding proteins.
This method is also applicable for other tissue types.
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Fluorescence In Situ Hybridization to Localize Transgenes in Plant Chromosomes
Time of Update: 2021-01-20
The technique is sensitive enough to detect single transgene copies and can reveal information about the complexity of a transgene insertion site as well as identifying plants homozygous for the transgene.
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Hordeivirus Isolation and RNA Extraction
Time of Update: 2021-01-20
Hordeiviruses have a diverse host range; BSMV, PSLV, and ALBV primarily infect members of the graminae, and LRSV is known to infect several members of the dicots.
However, to date, PSLV and ALBV have not been shown to be seed-transmitted.
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Identification of Small Molecule Inhibitors of Tomato Bushy Stunt Virus Replication
Time of Update: 2021-01-20
The identification of small molecule inhibitors of plant virus RNA replication is useful to develop antiviral drugs and to dissect various steps in the replication process.
Moreover, small molecule inhibitors could be effective tools to study similarities in replication strategies of various RNA viruses.
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Growth of Ferns from Spores in Axenic Culture
Time of Update: 2021-01-20
In this chapter, a method by which many fern species can be successfully grown from spores in axenic culture will be described.
The method can be used for the production of mature sporophytes or to provide a controllable system for biosystematic studies of, or experimentation with, fern gametophytes ( 1 , 2 ).
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PCR Detection of Potato Cyst Nematode
Time of Update: 2021-01-20
It is important for growers to know which species is present in their land as this determines its subsequent use.
The two speciesGlobodera pallida andGlobodera rostochiensis can be differentiated using an allele-specificPCR .
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Plant Secondary Metabolism and Challenges in Modifying Its Operation: An Overview
Time of Update: 2021-01-20
Plants have metabolic pathways leading to tens of thousands of secondary products capable of effectively responding to stress situations imposed by biotic and abiotic factors.
Ontogeny and circadian clock-controlled gene expression are also important features of plant secondary metabolism, as are master regulatory transcription factors.
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Ginseng (Panax ginseng)
Time of Update: 2021-01-19
A. Meyer by introducing new genetic materials and altering metabolic pathways that regulating the production of secondary compounds.
ginseng , production of transgenic plants viaAgrobacterium tumefaciens is performed via somatic embryogenesis using cotyledon or embryogenic callus as explants.
tumefaciens mediated-genetic transformation inP.
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Selection of Media for Tissue and Cell Culture
Time of Update: 2021-01-19
This chapter deals with the selection of growth media for the induction of callus and initiation of liquid suspension cultures from plants.
Although the first attempts at initiating cultures of plant cells were made by the German botanist G.
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RescueMu Protocols for Maize Functional Genomics
Time of Update: 2021-01-19
RescueMu is a modifiedMu1 transposon transformed into maize to permit mutagenesis and subsequent recovery of mutant alleles by plasmid rescue.
Germinal insertions may result in a mutant phenotype, andRescueMu permits recovery of 5–25 kb of transposon-flanking genomicDNA without having to construct and screen genomic DNA libraries.
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Transformation Protocols for Broadleaved Trees
Time of Update: 2021-01-19
Plant genetic manipulation by transformation is the insertion ofDNA directly into the genome of a plant cell, and the regeneration of whole plants from such cells. This is brought about by human int
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Genome-Wide Identification of MicroRNAs in Medicago truncatula by High-Throughput Sequencing
Time of Update: 2021-01-19
Medicago truncatula has been used as a model plant to study functional genomics of legume plants.
These methodologies associated with identification and functional characterization of miRNAs may provide useful tools for readers to study miRNAs in plants in general andMedicago truncatula in particular.
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Luteovirus Isolation and RNA Extraction
Time of Update: 2021-01-19
Purification procedures are presented for isolating BYDV from a monocot and PLRV from a dicot.
A procedure has been described that was effective in purifying six different luteoviruses representing both subgroups, and from both monocot and dicot plant tissues ( 5 ).
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Pollen Electrotransformation in Tobacco
Time of Update: 2021-01-19
Numerous techniques have been developed to transfer genes into plants to create genetically engineered crops that can tolerate environmental stresses, and to improve productivity and quality. The sea
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Opium Poppy (Papaver somniferum)
Time of Update: 2021-01-19
The genetic transformation of opium poppy,Papaver somniferum , offers the opportunity to study the mechanisms involved in the regulation of benzylisoquinoline and morphinan alkaloid biosynthesis.
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Plant Cell Immobilization in Alginate and Polyurethane Foam
Time of Update: 2021-01-19
Plant cell culture has great potential as an alternative system for the production of phytochemicals normally extracted from whole plants, many of which have commercial value. Suspension cultures of
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Transposon Tagging Using Activator (Ac) in Maize
Time of Update: 2021-01-19
The propensity ofAc to move to linked sites provides an opportunity to overcome this limitation whenAc elements are distributed at regular intervals throughout the genome.
This chapter details the use of such a system in maize through simple genetic manipulations.
A detailed protocol is also provided to cloneDNA flankingAc insertions.
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Cytological Dissection of the Triticeae Chromosomes by the Gametocidal System
Time of Update: 2021-01-19
This Gc system is also effective in inducing chromosomal breakages in Triticeae chromosomes added to common wheat.
The induced aberrant chromosomes can be identified by chromosome banding and fluorescence in situ hybridization and can be established in common wheat as dissection lines.
