PB 4.

Distant Hybridization

INTRODUCTION

• Distant hybridization refers to the crossing between:

1. Two different species of the same genus, or
2. Two different genera of the same family.

• Such crosses are also called distant crosses or wide crosses.

• Purpose: Wide crossing has been used in the genetic improvement of crop plants.
○ It is an effective method to transfer desirable genes into cultivated plants from
related species and genera.

• Types of Distant Hybridization:

1. Interspecific hybridization
2. Intergeneric hybridization

INTERSPECIFIC HYBRIDIZATION

Crossing or mating between two different species of the same genus is referred to as
interspecific hybridization.
○ It is also called intrageneric hybridization.

Main Features of Interspecific Hybridization:

1. When to use:
It is used when the desirable character is not found within the species of a crop.

2. Method effectiveness:
It is an effective method of transferring desirable genes into cultivated plants from their
related cultivated or wild species.

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 3. Success in propagation:
Interspecific hybridization is more successful in vegetatively propagated species (e.g.,
sugarcane and potato) than in seed-propagated species.

4. Types of crosses:
Interspecific hybridization gives rise to three types of crosses:
○ (a) Fully fertile
○ (b) Partially fertile
○ (c) Fully sterile

5. Leads to introgression:
Interspecific hybridization can lead to introgression, which refers to the transfer of
genes from one species into the genome of another species.

1. Fully Fertile Crosses

• Definition: Interspecific crosses are fully fertile between species with complete
chromosomal homology.
○ Chromosomes in such hybrids pair normally during meiosis, resulting in fully fertile
F₁ plants.
Examples in Crops:
1. Cotton:
○ There are four cultivated species of cotton:
Gossypium hirsutum, G. barbadense, G. arboreum, and G. herbaceum.
○ Crosses:
 G. hirsutum (2n = 52) × G. barbadense (2n = 52) → F₁ plants are fully fertile.
 G. arboreum (2n = 26) × G. herbaceum (2n = 26) → F₁ plants are fully fertile.

2. Partially Fertile Crosses

• Definition: Partially fertile crosses occur between species differing in chromosome
number but sharing some common chromosomes.
○ The resulting F₁ plants are partially fertile and partially sterile.
Examples in Crops:
1. Wheat:
○ Wheat has three species types:
 Diploid (2n = 14)
 Tetraploid (2n = 28)
 Hexaploid (2n = 42)
○ Cross Example:
 Triticum aestivum (2n = 42) × T. durum (2n = 28) → F₁ hybrids are partially
fertile.
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 fertile.
□ Chromosomes of A and B genomes are common.
□ Meiosis: 14 bivalents + 7 univalents → 2n = 35.

3. Fully Sterile Crosses

• Definition: Fully sterile crosses occur between species lacking chromosomal homology.
○ Chromosomes do not pair during meiosis, and F₁ plants are fully sterile.
○ Sterility can be overcome by chromosome doubling using colchicine treatment.
Examples in Crops:
1. Tobacco:
○ Clausen and Goodspeed (1928) crossed:
 Nicotiana sylvestris (2n = 24) × N. tomentosa (2n = 24) → F₁ was sterile.
 Colchicine treatment produced a tetraploid (2n = 48) → fertile plants
resembling N. tabacum.
○ Further cross:
 N. paniculata (2n = 24) × N. undulata (2n = 24) → F₁ was sterile.
 Colchicine treatment resulted in fertile amphidiploid (2n = 48) similar to N.
rustica.

INTERGENERIC HYBRIDIZATION

Intergeneric hybridization refers to crossing between two different genera of the same
family.
• Such crosses are rarely used in crop improvement due to various problems associated
with them.

Main Features:

1. Purpose: Used when desirable genes are not found in species of the same genus.
2. Limited use: Rarely used for specific character transfer into cultivated species.
3. Propagation: Generally applied in asexually propagated species.
4. Sterility:
○ F₁ hybrids between two genera are always sterile.
○ Fertility is restored by chromosome doubling using colchicine treatment.
5. Examples: Used by researchers to develop new crop species.

Examples of Intergeneric Hybridization

1. Wheat-Rye Cross

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 • First intergeneric cross in the Gramineae family:
○ Bread wheat (Triticum aestivum, 2n = 42) × Rye (Secale cereale, 2n = 14).
○ Carried out by Rimpau in 1890 (Sweden).
• Result:
○ F₁ plants were sterile, fertility restored through colchicine treatment.
○ Produced a fertile amphidiploid (2n = 56) named Triticale.
○ Significance:
 Combines the yield potential of wheat and the hardiness of rye.
 Triticale is grown commercially in Canada and Argentina.
 Research continues at CYMMIT, Mexico.

2. Radish-Cabbage Cross

• Cross:
○ Radish (Raphanus sativus) × Cabbage (Brassica oleracea) of the Cruciferae family.
○ Performed by Karpechenko in 1928 (Russia).
• Objective: To combine radish roots with cabbage leaves.
• Result:
○ F₁ plants were sterile.
○ Fertility restored by colchicine treatment → produced Raphanobrassica.
○ The hybrid had roots like cabbage and leaves like radish, which was a useless
combination.

1. Cross Incompatibility
• Meaning: Failure of two species to cross and produce hybrids.
• Cause: Reproductive barriers prevent fertilization.

2. Hybrid Inviability
• Meaning: Hybrids fail to survive or grow properly.
• Cause: Genetic differences lead to poor development and early death.

3. Hybrid Sterility
• Meaning: Hybrids are alive but cannot produce offspring.
• Cause: Chromosomes fail to pair correctly during meiosis, causing sterility.

4. Hybrid Breakdown
• Meaning: Hybrids may survive and reproduce, but future generations show reduced
fitness.
• Cause: Genetic incompatibility in later generations causes problems.

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ROLE OF WIDE CROSSING IN CROP IMPROVEMENT

• Wild species or wild genetic resources are potential sources of desirable genes for
various characters of crop plants.

Significance of Wide Crossing:

1. Wide crossing is an effective method to exploit desirable characters from wild species
for the improvement of cultivated crop plants.
2. Key Roles in Crop Improvement:
Wide crossing has played a significant role in:
○ (i) Improving crop plants for:
 Disease and insect resistance
 Quality
 Adaptation
 Yield
 Mode of reproduction
 Other specific characters
○ (ii) Developing commercial hybrids in some crops.
○ (iii) Creation of new crops.

1. Character Improvement

(i) Disease and Insect Resistance:

• Distant hybridization helps transfer disease resistance from wild species to cultivated
ones.
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 ones.
○ Examples:
 Resistance to rust
 Resistance to black arm in cotton
 Resistance to mosaic virus, wildfire, and Fusarium wilt
(ii) Improvement in Quality:
• Wild species improve quality traits in crops.
○ Examples:
 Protein content in rice, oats, and rye
 Fibre length in cotton
 Oil quality in oil palm

2. Hybrid Varieties
• Improved hybrid cultivars have been developed using wild species, especially in:
○ Sugarcane
○ Potato
○ Forage crops
• Examples include successful upland cotton varieties (MCU 2, MCU 5) and hybrids like
Pearl millet × Napier grass for superior fodder quality.

3. New Crop Species

• Distant hybridization and polyploidy have created new crop species.
○ Examples:
 Triticale: A crop from an intergeneric cross between Triticum aestivum and
Secale cereale.
 Combines the yield potential of wheat and the hardiness of rye.

Limitations of Wide Crossing

1. Crossing Problems:
○ Issues like cross incompatibility, hybrid inviability, sterility, and breakdown make
hybridization difficult.
2. Special Techniques Needed:
○ Methods like ploidy manipulation, grafting, and embryo culture are required,
making it complex.
3. Linked Undesirable Traits:
○ Desirable traits are often linked to undesirable characters, which limits their use.
4. Undesirable Hybrids:
Hybrids may show non-flowering, late maturity, or useless traits like
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 ○ Hybrids may show non-flowering, late maturity, or useless traits like
Raphanobrassica.
5. Recessive Gene Transfer:
○ Transferring traits controlled by recessive genes is very difficult in hybrids.
6. Complex Character Transfer:
○ Character transfer in distant hybridization is not as simple as in close crosses.

Synthetic Varieties And Composite Varieties

A synthetic variety is developed by:
○ Intermating in all possible combinations between a number of inbred lines or
individuals.
○ Mixing the seeds of F₁ crosses in equal quantity.
○ The resulting variety is maintained by open pollination.

• Key Features:
• Relevance:
○ Synthetic varieties are ideal for cross-pollinated crops like maize, pearl millet, and
alfalfa.
• Base Material:
○ Derived from inbreds, clones, and open-pollinated varieties.
○ Typically involves 5-8 inbred lines with good general combining ability (GCA).
• Genetic Concept:
○ Synthetic varieties exploit heterosis (hybrid vigor).
○ Only partial heterosis is retained due to continuous open pollination.
○ They rely more on additive gene action.
• Genetic Constitution:
○ Initially, synthetic varieties consist of heterozygotes.
○ Over generations, due to selfing, they include a mix of homozygotes and
heterozygotes, making them heterogeneous.
• Adaptation:
○ Synthetic varieties are polymorphic and stable, adapting well to environmental
variations.
○ They provide stable yields under fluctuating conditions.
• Disease Resistance:
○ Due to their genetic diversity, synthetic varieties show better resistance to plant
diseases.

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 diseases.
• Reconstitution:
○ Can be safely grown for 4-5 years without a drop in yield potential.
○ After five years, it’s recommended to reconstitute the variety to maintain
productivity.
• Yield Level:
○ Higher than open-pollinated varieties, but lower than single or double-cross
hybrids.
○ Advantage: Seeds are much cheaper compared to hybrids.
• Designation:
○ Generations of synthetic varieties are labeled as Syn₀, Syn₁, Syn₂, and so on.

Steps in Development of Synthetic Variety

1. Isolation of Inbreds:
○ Materials such as inbred lines, clones, or open-pollinated varieties are selected.
○ Recurrent selection methods are used to improve the inbreds.
○ Inbred lines with one generation of selfing can be used for synthetic variety
development.
○ If inbred lines or clones are the parents, the variety can be reconstituted.
○ Short-term inbreds cannot be reconstituted because they are heterozygous for many
genes.

1. Evaluation of Inbreds for GCA (General Combining Ability):

○ Inbred lines are tested for their ability to combine well with others.
○ There are three methods for evaluating inbreds:
1. Top cross method:
□ Inbreds are crossed with a common tester. Progeny is evaluated for yield
and performance.
2. Poly cross method:
□ Selected inbreds intermated by open pollination and progeny tested.
3. Single cross method:
□ All possible single crosses are made between inbreds, evaluated for yield
and general combining ability.
○ The first two methods are used to evaluate large numbers of inbreds, while the
single cross method evaluates only a small number due to time limitations.

1. Intermating of Good General Combining Inbreds:

○ Selected inbreds with superior GCA are crossed in all possible combinations.
If there are n inbreds, the total crosses are given by the formula: n(n−1)/2n(n-1)/2
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 ○ If there are n inbreds, the total crosses are given by the formula: n(n−1)/2n(n-1)/2
Example: For 6 inbred lines → 15 crosses.
○ Seeds of each cross are mixed in equal quantity to develop a synthetic variety.

1. Mixing of F₁ Seeds:
○ The seeds of all F₁ crosses are mixed in equal amounts.
○ This variety is maintained by open pollination for one or two generations:
 Generations are labeled as Syn₁, Syn₂, and so on.
○ After stabilization, the synthetic variety is distributed to farmers for cultivation.

Merits and Demerits of Synthetic Varieties

Merits
1. Exploitation of Heterosis:
○ Synthetic varieties use heterosis (hybrid vigor) even in species where producing
hybrid seeds is difficult.
2. Cost-Effective:
○ Seeds of synthetic varieties are cheaper than single or double cross hybrids.
○ Small farmers can afford these seeds.
3. Adaptability:
○ They are more adaptable to environmental changes due to their broad genetic base
and variability.
4. Genetic Variability:
○ Synthetic varieties show vast genetic variability, giving them better disease
resistance.
5. Seed Reuse:
○ Farmers do not need to purchase fresh seed every year. They can use their own-
saved seeds for 4-5 years.

Demerits
1. Less Uniform and Attractive:
○ Produce is less uniform and less attractive compared to hybrids due to genetic
variability.
2. Lower Yield:
○ Yield is generally poorer than single or double cross hybrids because only partial
heterosis is exploited.
3. Limited to Cross-Pollinated Crops:
○ Synthetic varieties are used only for cross-pollinated species, while hybrids can be
developed for both cross and self-pollinated species.

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 Composite Varieties
Composite varieties are developed by mixing genotypes from several sources. These
genotypes are selected based on similar traits like maturity, height, and seed size, and
the variety is maintained by open pollination.

Key Features of Composite Varieties

1. Base Material:
○ Genotypes with similar maturity, seed size, color, and height are mixed.
○ Seeds are maintained by open pollination.
○ Farmers can use these seeds for 3-4 years.
2. Genetic Concept:
○ Composite varieties make use of heterosis (hybrid vigor) in cross-pollinated crops.
3. Adaptation:
○ Composite varieties are more stable and highly adaptable to environmental
changes.
4. Reconstitution:
○ Composite varieties are derived from heterozygous genotypes, so they cannot be
reconstituted due to changes in gene frequency over time.

Steps in Development of Composite Varieties

1. Selection of Base Material:

○ Use open-pollinated varieties or heterozygous sources with similar traits like
maturity, height, seed size, and seed color.
2. Intermating Selected Genotypes:
○ Selected genotypes are crossed in all possible combinations.
○ Formula: n(n−1)/2n(n-1)/2 → For 10 genotypes, 45 single crosses are made.
3. Evaluation of Crosses:
○ Single crosses are evaluated in replicated trials for yield performance.
○ Generations F₂, F₃, and F₄ are checked for yield and little to no inbreeding
depression.
4. Mixing Parental Seeds:
○ Seeds of superior crosses are mixed in equal quantity.
○ Open pollination is carried out for one or two generations to stabilize the variety.

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 ○ Composite varieties exploit additive gene action and minimize inbreeding
depression.

Somaclonal Variations
.
Genetic variations in plants produced during plant tissue culture, detected as genetic or
phenotypic traits.

Basic Features:
• Often involve chromosomal changes such as altered leaf shape, growth rate, and
fertility.
• Heritable mutations persist even after planting in the field.

Mechanism:
1. Genetic (Heritable) Variations:
○ Pre-existing in somatic cells.
○ Caused by mutations and other DNA changes.
○ Occur at high frequency.
2. Epigenetic (Non-heritable) Variations:
○ Generated during tissue culture.
○ Temporary changes in phenotype.
○ Occur at low frequency.

Steps Involved:
1. Callus tissue formation
2. Organogenesis (development of organs).
3. Regeneration and hardening of plants.
4. Selfing and selection of somaclonal variants.

Causes:
1. Physiological Causes:
○ Exposure to plant growth regulators.
○ Culture conditions.
2. Genetic Causes:
○ Chromosome Number Changes:
 Aneuploidy, polyploidy, translocation, inversion.
Chromosome Structure Changes:

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 ○ Chromosome Structure Changes:
 Deletion, duplication, inversion, translocation.
○ Gene Mutations:
 Transition, transversion, insertion, deletion.
○ DNA Sequence:
 Changes detected by restriction enzymes.
 Altered proteins or DNA methylation.
3. Biochemical Causes:
○ Changes in carbon metabolism, starch biosynthesis, nitrogen metabolism, and
antibiotic resistance.

Detection and Isolation of Variants:

1. Morphological Analysis:
○ Qualitative traits (plant height, flowering date).
○ Quantitative traits (yield, seed content).
2. Cytological Studies:
○ Staining root/leaf tips to observe chromosome morphology.
3. DNA Content Detection:
○ Measuring DNA content using Cytophotometry.
4. Gel Electrophoresis:
○ Detect changes in enzymes, proteins, pigments, or alkaloids.
5. Disease Resistance Detection:
○ Using pathogens/toxins as selection agents.
6. Herbicide Resistance Detection:
○ Adding herbicide to cultures to select resistant plants.
7. Environmental Stress Tolerance Detection:
○ Salt, waterlogging, drought, temperature, and mineral toxicity resistance.

Advantages:
1. Crop Improvement:
○ Adds new genetic variations.
○ Improves resistance to toxins, herbicides, and mineral stress.
2. Improved Production:
○ Increased production of secondary metabolites.
3. Tree Breeding:
○ Suitable for tree species improvement.
4. Specific Examples:
○ Sugarcane: Higher yield, disease resistance.
○ Potatoes: Yield improvement.
○ Geraniums: Better scent varieties.
○ Paulownia: Leaf variegation.

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Disadvantages:
1. Not suitable where clonal uniformity is needed.
2. Results can be undesirable or genetically unstable.
3. Requires extensive field trials.
4. Ineffective for complex traits like yield or quality.
5. May cause pleiotropic effects (multiple unintended changes).

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