Plant Breeding and Plant Improvement –

Conventional breeding methods are the most widely used for crop improvement. But in certain
situations these methods have to be supplemented with plant tissue culture techniques, either to
increase their efficiency or to achieve an objective not possible through conventional methods.

Embryo culture is now routinely used in recovery of hybrid plants from distant crosses. Some
examples are recovery' of hybrids from Hordeum vulgare x Agropyron repens and H. vulgare x
Triticum sp. In the case of Triticale, a rare hybrid between Triticum and Secale develops viable
seeds. But most of the tetraploid and hexaploid wheat carry two dominant genes, Kr1 and Kr2,
which prevent seed development in crosses with Secale.

The hybrid seeds are minute, poorly developed and show very poor germination. By embryo
culture, 50-70% hybrid seedlings have been obtained. Hybrid seedlings from T. aestivum x H.
vulgare are not obtained. But in embryo culture when H. vulgare or T. aestivum (used as male) is
crossed with H. bulbosum (used as female), the chromosome complement of H. bulbosum is
eliminated from the developing embryo.

Most of the seedlings obtained from such crosses are haploid, having only one set of
chromosomes, either from the H. vulgare or the T. aestivum parent. Embryo culture is also useful
for propagation of orchids, shortening the breeding cycle and overcoming seed dormancy.

In meristem culture the shoot apical meristem along with some surrounding tissue is grown in
vitro. This is used for clonal propagation and recovery of virus free plants and is potentially
useful in germplasm exchange and long term storage of germplasm through freeze preservation.

Anther and pollen culture has potential application in a plant breeding and plant improvement
program for the production of haploid as well as homozygous diploid plants. Year round rapid
clonal propagation using plant tissue culture techniques has highlighted possibilities for new
plant improvement techniques. Protoplast culture and somatic hybridization is a promising line
for plant breeding and plant improvement techniques.

Another important approach is the mutation of tissue culture cells to produce a mutant line from
which plants can be raised. Production of a mutant line is highly desirable for plant breeding.
Callus cells, produced either from a vegetative cell or reproductive tissues, can be subjected to a
range of mutagenic chemicals, e.g. N-nitroso-N-methyl urea, or ionizing radiations, e.g. gamma
rays.,

The hope is that such treatment will effect permanent changes in the DNA pattern of some cells.
Plants could be raised from the treated cultures and any mutant whole plants could be selected
from the population either by physical differences or by metabolic/biochemical differences.
Biochemical mutants could be selected for disease resistance, resistance to phytotoxin,
improvement of nutritional quality, adaptation of plants to stress conditions, e.g. saline soils, and
to increase the biosynthesis of plant products used for medicinal or industrial purposes.
Modern plant breeding
Modern plant breeding uses techniques of molecular biology to select, or in the case of genetic
modification, to insert, desirable traits into plants.

Modern facilities in molecular biology has converted classical plant breeding to molecular plant
breeding

Steps of Plant Breeding

The following are the major activities of plant breeding;

1. Creation of variation
2. Selection
3. Evaluation
4. Release
5. Multiplication
6. Distribution of the new variety

Marker assisted selection

Sometimes many different genes can influence a desirable trait in plant breeding. The use of
tools such as molecular markers or DNA fingerprinting can map thousands of genes. This allows
plant breeders to screen large populations of plants for those that possess the trait of interest. The
screening is based on the presence or absence of a certain gene as determined by laboratory
procedures, rather than on the visual identification of the expressed trait in the plant.

Reverse Breeding and Doubled Haploids (DH)

A method for efficiently producing homozygous plants from a heterozygous starting plant, which
has all desirable traits. This starting plant is induced to produce doubled haploid from haploid
cells, and later on creating homozygous/doubled haploid plants from those cells. While in natural
offspring genetic recombination occurs and traits can be unlinked from each other, in doubled
haploid cells and in the resulting DH plants recombination is no longer an issue. There, a
recombination between two corresponding chromosomes does not lead to un-linkage of alleles or
traits, since it just leads to recombination with its identical copy. Thus, traits on one chromosome
stay linked. Selecting those offspring having the desired set of chromosomes and crossing them
will result in a final F1 hybrid plant, having exactly the same set of chromosomes, genes and
traits as the starting hybrid plant. The homozygous parental lines can reconstitute the original
heterozygous plant by crossing, if desired even in a large quantity. An individual heterozygous
plant can be converted into a heterozygous variety (F1 hybrid) without the necessity of
vegetative propagation but as the result of the cross of two homozygous/doubled haploid lines
derived from the originally selected plant. patent
Genetic modification

Genetic modification of plants is achieved by adding a specific gene or genes to a plant, or by

knocking down a gene with RNAi, to produce a desirable phenotype. The plants resulting from
adding a gene are often referred to as transgenic plants. If for genetic modification genes of the
species or of a crossable plant are used under control of their native promoter, then they are
called cisgenic plants. Genetic modification can produce a plant with the desired trait or traits
faster than classical breeding because the majority of the plant's genome is not altered.

To genetically modify a plant, a genetic construct must be designed so that the gene to be added
or removed will be expressed by the plant. To do this, a promoter to drive transcription and a
termination sequence to stop transcription of the new gene, and the gene or genes of interest
must be introduced to the plant. A marker for the selection of transformed plants is also included.
In the laboratory, antibiotic resistance is a commonly used marker: Plants that have been
successfully transformed will grow on media containing antibiotics; plants that have not been
transformed will die. In some instances markers for selection are removed by backcrossing with
the parent plant prior to commercial release.

The construct can be inserted in the plant genome by genetic recombination using the bacteria
Agrobacterium tumefaciens or A. rhizogenes, or by direct methods like the gene gun or
microinjection. Using plant viruses to insert genetic constructs into plants is also a possibility,
but the technique is limited by the host range of the virus. For example, Cauliflower mosaic virus
(CaMV) only infects cauliflower and related species. Another limitation of viral vectors is that
the virus is not usually passed on the progeny, so every plant has to be inoculated.

The majority of commercially released transgenic plants are currently limited to plants that have
introduced resistance to insect pests and herbicides. Insect resistance is achieved through
incorporation of a gene from Bacillus thuringiensis (Bt) that encodes a protein that is toxic to
some insects. For example, the cotton bollworm, a common cotton pest, feeds on Bt cotton it will
ingest the toxin and die. Herbicides usually work by binding to certain plant enzymes and
inhibiting their action. The enzymes that the herbicide inhibits are known as the herbicides
target site. Herbicide resistance can be engineered into crops by expressing a version of target
site protein that is not inhibited by the herbicide. This is the method used to produce glyphosate
resistant crop plants (See Glyphosate)

Genetic modification of plants that can produce pharmaceuticals (and industrial chemicals),
sometimes called pharmacrops, is a rather radical new area of plant breeding.
Domestication to Crop Improvement: Genetic Resources for Sorghum
and Saccharum (Andropogoneae)

1. Sally L. Dillon2,
2. Frances M. Shapter1,
3. Robert J. Henry1,*,
4. Giovanni Cordeiro1,
5. Liz Izquierdo1 and
6. L. Slade Lee1

Oxford Journals,Life Sciences, Annals of Botany Volume100, Issue5 Pp. 975-989.

The role of genomics in improving domesticated S. bicolor

Sorghum bicolor, a diploid, has a relatively small genome (735 Mbp), which although larger than
rice (389 Mbp) is smaller than the other important cereals (wheat 16 900 Mbp, maize 2600
Mbp). The last genome duplication event for the S. bicolor genome seems to have occurred
much earlier than the divergence of the major cereal crops from a common ancestor (Paterson et
al., 2004). Completion of the whole genome sequencing project in 2007 will exponentially
increase the sequence data available for Sorghum and will provide valuable information on
cereal domestication in the African continent, an event that appears to have occurred
independently of other continents though by similar reinforced selective pressures (Paterson et
al., 2004). In a way, the sorghum genome sequencing will close a biographic triangle into the
knowledge of the polymorphism shared before the divergence of these important grasses and
ultimately in the understanding of the evolution in cereals crops between Africa, America and
Asia (Kresovich et al., 2005). The tenets of colinearity and microlinearity of grass genomes
mean that our knowledge of other cereals and their evolutionary ties will also greatly improve.
Due to their economic and scientific value, cereal genomes have been studied over the last 15
years using highly advanced technologies. The similarity at the DNA level makes it possible to
use comparative genetics to look for particular genes of unknown sequence between the genomes
with the aim of using that information to develop new varieties or discovering new genes that
could have a potential impact on traits that are of global importance (e.g. food quality, drought
resistance).

The genetic diversity existing within and between Australian Sorghum species was recently
evaluated using simple sequence repeats (SSRs) (Dillon et al., 2005). SSRs were sourced from
the cultivated S. bicolor (Brown et al., 1996; Taramino et al., 1997; Kong et al., 2000) to
determine diversity in these closely related taxa. This method has successfully evaluated
diversity in the related species of many crop groups (e.g. Peakall et al., 1998; Hernández et al.,
2001; Chen et al., 2002; Scott et al., 2003; González-Martínez et al., 2004; Sudupak, 2004). This
evaluation of the Australian species has shown significantly higher levels of genetic diversity
both between (inter-) and within (intra-) species compared with the intra-specific diversity of S.
bicolor varieties. The relatively high transfer rate of S. bicolor-derived SSRs to the wild species
and their high level of diversity suggests that these SSRs are an efficient, highly informative
source of molecular markers for the undomesticated Sorghum species.