植物生理

Plant single cell sampling and analysis allows the determination of solute concentrations in individual cells and tissues. This is particularly important when studying mineral nutrition, where the cell- and tissue-specific distribution of individual mineral nutrients incr ...

Xylem and phloem are essential for the exchange of solutes and signals among organs of land plants. The synergy of both enables the transport and ultimately the partitioning of water, nutrients, metabolic products and signals among the organs of plants. The collection and analysis of xylem sap a ...

The transport tubes of the phloem are essential for higher plants. They not only provide the route for the distribution of assimilates produced during photosynthesis from source to sink organs but also (re-) distribute mineral nutrients. Additionally, the phloem is essential for sending ...

Noninvasive microelectrode ion flux measurements (the MIFE™ technique) allow the concurrent quantification of net fluxes of several ions with high spatial (several μm) and temporal (ca 5 s) resolution. The MIFE technique has become a popular tool for studying the adaptive responses of pl ...

Standard procedures for the tracing of ion fluxes into roots of plants are described here, with emphasis on cations, especially potassium (K+). We focus in particular on the measurement of unidirectional influx by use of radiotracers and provide a brief introduction to compartmental anal ...

Herbicide resistance screening is a method that can be used not only to determine presence of the enzyme, phosphinothricin acetyltransferase, encoded by either the Bar or the Pat gene in transgenic maize, but also to assess the inheritance ratio of those genes in a segregating population. Herb ...

The analysis of transgene inheritance is an important step in the molecular and genetic characterization of transgenes. In this manuscript, two approaches to characterize the inheritance of transgenes are described. The first approach is based on the expression of the transgene phen ...

Backcross breeding enables breeders to transfer a desired trait such as a transgene from one variety (donor parent, DP) into the favored genetic background of another (recurrent parent, RP). If the trait of interest is produced by a dominant gene, this process involves four rounds of backcrossi ...

Transformation with Agrobacterium tumefaciens is the preferred method for delivery of transgenes into a wide range of plant species including maize. Optimized protocols for the Agrobacterium-mediated transformation of freshly isolated immature embryos and embryogenic T ...

Biolistic gun-mediated transformation is one of the two most effective and popular methods for introducing genes into maize. In this chapter, we describe a detailed protocol for genetic transformation of the maize genotype, Hi II. The protocol uses 0.6-μm gold particles as microcarriers a ...

Transient expression is the nonstable expression of a transgene whereby the transgene does not integrate into the host’s genomic DNA. Transient expression assays have 20 years of history in plant molecular biology research, being used to answer a variety of questions. The method described ...

The first step of any maize transformation project is to select gene expression elements that will make up an effective construct. When designing a gene construct, one must have a full understanding of the different expression elements that are currently available and of the strategies that h ...

The use of fluorescent proteins to localize gene products in living cells has revolutionized cell biology. Although maize has excellent genetics resources, the use of fluorescent proteins in maize cell biology has not been well developed. To date, protein localization in this species has m ...

There has been rapid progress in recent years in extending gene transfer capabilities to include plant species that fall outside the normal host range of Agrobacterium. Methods that allow direct DNA delivery into plant cells have contributed significantly to this expanded capability. ...

RNAi can be an effective means to regulate endogenous gene expression in maize and as such represents an important reverse genetics tool. This approach involves designing a transgenic construct that creates a double stranded RNA (dsRNA) upon transcription, and introducing the transg ...

The transgenic RescueMu lines were designed for and successfully used in our maize gene discovery project. The pBluescript-containing RescueMu transposon can be readily recovered by a procedure called plasmid rescue. Plasmid rescue is a technique for recovering bacterial plasm ...

Efficient methods to characterize transgenic plants are important to quickly understand the state of the transformant. Determining transgene copy number is an important step in transformant characterization and can differentiate between complex and simple transformati ...

This method allows high-throughput qualitative screening to identify seeds containing a transgene product in endosperm tissue. It is particularly useful for determining genetic segregation ratios or identifying seeds to be advanced in a breeding program. Tissue printing is used ...

Sensitive and reproducible genotyping tools are fundamental in interpreting and substantiating genetic data. In cases where alternative assays like PCR are not applicable, a sensitive genomic Southern protocol is needed. Our maize gene discovery work using the RescueMu transge ...

There are thousands of porphyrins and metalloporphyrins, and hundreds of new derivatives appear each year. This variety arises because the cyclic conjugated tetrapyrrole nucleus (Figure 1) can have different substituents at the eight β-pyrrole positions, at the four meso carbon ato ...