植物学技术

For stable transformation of cereals through PEG-mediated DNA uptake into protoplasts, the two most critical requisites are the ability to isolate and culture protoplasts in large numbers, and the development of an efficient and reliable system for routine plant regeneration from pr ...

The plastid genome (or “plastome”) encodes a number of proteins associated with the structure and function of chloroplasts, as well as tRNAs and rRNAs associated with the plastid translational machinery (1,2). Although there have been numerous studies on the genetics of algal chloroplas ...

The prospect of utilizing plant tissue cultures as a means of generating and identifying novel genetic variants is one that has sparked the interest of researchers for many years. It has resulted in large numbers of research papers describing the selection and characterization of mutants, a ...

Agrobacterium-mediated transformation of plants is now applicable to many dicotyledonous and also several monocotyledonous plant species. It can be used to transform many different species based on various factors: the broad host range of Agrobacterium (1), the regeneration res ...

Crosses between species of the same or different genera are called wide crosses (WCs). They are useful in plant improvement for gene transfer and haploid production, and in general biology, for genome mapping, study of chromosome behavior, and phylogenetic relationships (1–8). Broadeni ...

Microspores have two developmental options when they are isolated from anthers and cultured in vitro. In a rich medium, they can develop into mature pollen grains that are fertile on pollination in vivo (1,2). Their development closely resembles pollen formed in vivo, whereas the in vitro cultu ...

The establishment of true breeding lines is a critical step for variety development in most crop plants. Traditionally, plant breeders have achieved homozygosity for all genes by using the time- and labor-intensive methods of self-fertilization or backcrossing. The phenomenon of par ...

Statistics and experimental design are important tools for the plant biotechnologist and should be used when planning and conducting experiments as well as during the analysis and interpretation of results. This chapter provides some basic concepts important to the statistical an ...

Plant tissue culture, or the aseptic culture of cells, tissues, organs, and their components under defined physical and chemical conditions in vitro, is an important tool in both basic and applied studies as well as in commercial application. It owes its origin to the ideas of the German scientist, H ...

Somatic embryogenesis (SE) provides a useful model to study embryo development in plants. In contrast to zygotic embryogenesis, SE can easily be observed, the culture conditions can be controlled, and large quantities of embryos can be easily obtained. In Coffea spp several model systems ha ...

Encapsulation-vitrification, which is a combination of encapsulation-dehydration and vitrification procedures, is a newly developed technique for cryopreservation of plant germoplasm. Here, we describe the protocol of this methodology, using grapevine (Vitis) as a model p ...

Callus and cell suspension can be used for long-term cell cultures maintenance. This chapter describes procedures for the induction of somatic embryos of garlic, keeping a regeneration capacity for more than 5 yr, as well as the maintenance of a tobacco suspension culture (NT-1 cells), for more t ...

Generation of somatic embryos in spruce is achieved through the execution of five steps designated as: (1) induction of embryogenic tissue, (2) maintenance of embryogenic tissue, (3) embryo development, (4) embryo maturation, and (5) conversion into plants. Depending on species and geno ...

An overview of the methods for assessing cell viability in in vitro cultures is presented. The protocols of four of the most commonly used assays are described in detail, so the readers may be able to determine which assay is suitable for their own projects using plant cell cultures.

A new type of bioreactor system for plant micropropagation is described that incorporates a number of features specifically designed to simplify its operation and reduce production costs. The BioMINT unit is a mid-sized (1.2 L) reactor that operates on the principle of temporary immersio ...

In cassava both direct and indirect somatic embryogenesis is described. Direct somatic embryogenesis starts with the culture of leaf explants on Murashige and Skoog (MS) medium supplemented with auxins. Somatic embryos undergo secondary somatic embryogenesis when cultured on t ...

This chapter presents a protocol that will show ways to obtain photoautotrophic coconut in vitro plants and outlines protocol for improving photosynthesis and field performance. This protocol involves reducing sucrose concentration from the growing medium while simultaneo ...

Procedures to isolate native light-harvesting chlorophyll-a/b complex (LHCIIb) and to reconstitute recombinant LHCIIb are described. Separation of trimeric from monomeric forms and free pigment by sucrose density-gradient ultracentrifugation can be applied to both nati ...

In this chapter we describe isolation of loosely stacked lamellar aggregates of LHCII that are capable of undergoing light-induced reversible structural changes, similar to those in granal thylakoid membranes (LHCII, the main chlorophyll a/b light-harvesting antenna complex of ...

Chorophylls and carotenoids are functionally important pigment molecules in photosynthetic organisms. Methods for the determination of chlorophylls a and b, β-carotene, neoxanthin, and the pigments that are involved in photoprotective cycles such as the xanthophylls are dis ...