植物生理

The Furovirus (fungus-transmitted rod-shaped) group, whose type member is soil-borne wheat mosaic virus (SBWMV) (1), is a very heterogeneous group infecting a wide variety of hosts. The group consists of viruses that are naturally transmitted by soil-borne fungal vectors, generally of t ...

Agrobacterium-mediated transfer of cloned DNA copies of infectious viral genomes to plant tissues has been termed “agroinoculation” or “agroinfection” (for review, see ref. 1). The technique has been used successfully for a wide variety of virus groups, including the caulimoviruses a ...

Introduction of foreign genes into plant cells can be achieved by a variety of methods, including direct transfer into protoplasts using chemical (1–3; see also Chapter 7, this volume) and electrical methods (4–6). These methods can be used to study genes both transiently (1, 5,7) and when stably int ...

The cereals are one of the groups of crops more recalcitrant to transformation. Since, with the exception of rice (1), cereals have not been transformed by Agrobacterium, and highly regenerable protoplast systems are difficult to obtain, these species have remained untransformed for a much ...

The chloramphenicol acetyltransferase (cat) gene, isolated from transposon Tn9 of Escherichia coli (1), is a convenient genetic marker for studies of transformation. (For a detailed description of the CAT gene, see Chapter 1.) In plants, the CAT system is generally used in transient, rather t ...

The stable introduction of genes into plants through genetic engineering normally necessitates the use of a selectable marker, especially when the transformation frequency is low (e.g., 1.0 � 10−3 to 10−6). Marker genes enable quantification of both the transformation efficiency and ge ...

The study of how genes are organized within the genome is a major step in the characterization of cloned DNA sequences and is important in the analysis of plant transformation. In both cases, accurate information is required about the number of copies of a given DNA sequence and their location in the gen ...

Today the isolation and characterization of a gene of interest from any organism has become a standard procedure. One of the milestones that have facilitated this procedure in particular, has been the development of in vitro techniques for amplification of specific RNA or DNA sequences. By far ...

Eukaryotic messenger RNA (mRNA) can be separated from the other RNA species in a total RNA preparation by affinity chromatography by virtue of the presence of a polyadenylic acid “tail,” 20–25 bases long, at the 3′ end of the molecule (1). Oligo(dT)-cellulose is used routinely for homemade affinity c ...

Efficient extraction of high quality RNA from a variety of plant tissues is an important first step in many procedures, such as analysis of gene expression, cDNA library construction, and in vitro translation. The procedure described here, which is essentially the same as described in Draper et ...

Soon after the introduction of the Southern blotting procedure (1), an analogous technique, known as northern blotting, was developed for the analysis of RNA sequences (2). Both procedures rely on the annealing (hybridization) of complementary single-stranded nucleic acid molecul ...

Nuclear “run-on” (or “run-off”) transcription assays have been used to obtain quantitative information about the relative rates of transcription of different genes in nuclei isolated from a particular tissue or organ. This information can then be used in analyzing the factors that contr ...

RNase A/T1 mapping provides a sensitive and quantitative method of expression analysis of known gene sequences. It differs from primer-derived analyses such as primer extension and reverse transcriptase-PCR by the use of a probe that is colinear with the transcript under study. This prot ...

A number of protocols are available for the analysis of RNA transcripts: Northern analysis (see Chapter 18), S1 mapping, primer extension, RNase AT1 mapping (see Chapter 20), and the more recent reverse transcriptase/PCR amplification (see Chapters 22 and24). These protocols have been app ...

Class II gene in vitro transcription systems from animal cells (1–3) or fungi (4) have been essential for elucidating the mechanisms regulating gene expression. Crude protein extracts from Hela cell nuclei that allow accurate transcription of class II promoters in vitro have led to the inves ...

Reverse transcriptase-polymerase chain reaction (RT-PCR) is a highly sensitive method that permits the detection and quantification of RNA transcripts. It is particularly useful where transcripts are in very low abundance and where the amount of starting RNA is fairly limited, such as ...

As described in previous chapters (Chapters 15–23), techniques for the measurement of gene expression at the RNA level include RNA gel blot analysis, S1 mapping, primer extension, RNase protection, and, more recently, reverse transcription-polymerase chain reaction (RT-PCR) (1). This ...

Peanut or groundnut (Arachis hypogaea L.) is a member of the legume family, originated in South America, and today is largely cultivated in many tropical and subtropical areas worldwide. Peanut seeds are an important source of protein, carbohydrates, and oil for humans and animals. In addition to ...

The transfer of genetic material into soybean (Glycine max L. Merr.) plant tissues has been accomplished by several methods: electroporation (1), microprojectiles (2), and by the more widely used Agrobacterium-mediated T-DNA transfer (3,4). The transformation of soybean by electrop ...

Chlorogenic acid is a phenolic compound (Fig. 1). It is an antioxidant and a carcinogenic inhibitor; however, its presence in oilseeds and grains poses nutritional problems. Chlorogenic acid found in the range of 2 to 4 g/100 g of defatted sunflower meal is a matter of concern. Inclusion of high levels of s ...