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MONOSOMICS
Manjappa
Ph. D Scholar
Dept. of Genetics and Plant Breeding
1
Nutshell
• Introduction
• Methods of production
• Identification of monosomics
• Meiotic behavior of monosomics
• Breeding behavior
• Checking identity of monosomics
• Locating genes on polyploid crops
• Human disorder
2
Introduction
 Polyploids can tolerate not diploid (except Maize - diploid or
segmental polyploid)
History:
 Scattered work on monosomics initiated in early 1920’s
 Systematic work started by-
 Clausen & Cameron (1944)- Nicotiana tabacum
 Sears (1954)- Hexaploid wheat
 Endrizzi et al. (1963, 84, 85)- Gossypium hirsutum
3
Methods of Monosomic Production
1. From haploids:
4
2. Backcrosses of interspecific hybrids
5
Production of monosomics of D genome in bread wheat
6
3. From partially asynaptic plants
7
4. Irradiation treatment: Inflorescence is irradiated
 Non-disjunction of normal bivalents- production of gametes with
2x+1 and 2x-1
5. Spontaneous production:
 Occasional non-disjunction of bivalents during meiosis
8
Meiotic Nondisjunction Generates Aneuploidies
9
Description and identification of monosomics
• Generally polyploids don’t show drastic alterations in
morphology (bcz. addition of chromosome to diploid creates
greater disturbance)
• Chromosome count in mitosis & univalents in meiosis
N. Tabaccum x N. sylverstris
(Monosomic)
12I (T) + 11I (S) 12I (S)
12I (T) + 11II + 1I (S)
2n= 11II + 13I
Monosomic for Sylverstris genome
N. Tabaccum x N. sylverstris
(Monosomic)
11I (T) + 12I (S) 12I (S)
11I (T) + 12II + 1I (S)
2n= 12II + 11I
Not monosomic for Sylverstris
genome 10
Identification in Cotton
1. Cross monosomic with proginator
2. Study of meiosis in hybrids b/w monosomic
and translocated testers that are available in
cotton
11
12
Identification of monosomics & related phenomena in
wheat
a) Classification of monosomics in durum group (A & B
genome) & D genome
T. aestivum X T. durum
2n= 20II+1I (ABD) 2n= 14II (AB)
Meiosis: 14II + 6I
(AB) (D)
Monosomic in D genome
T. aestivum X T. durum
2n= 20II+1I (ABD) 2n= 14II (AB)
Meiosis: 13II + 8I
(AB) (D+A/B)
Monosomic in A/B genome
13
Distinction b/w monosomics of A & B genomes
• If telocentric chr. was involved in the formation of
heteromorphic bivalent, then the monosomic belongs to A
genome if not to B genome
Monosomics X Amphidiploid
(Ditelocentric) AADD
(2n= 20II + 1II telo) (2n= 14II)
F1: 34 + telo
14
C. Assessing chr. to homeologous groups in Wheat
(compensating nullisomic-tetrasomic lines
• It has been demo. that the presence of an extra chr. may separately
compensate phenotypically for the absence of each of the other
two chr. “This is an evidence of homeology among 3 chr.”
• Confirmed by-
1. Study at gametophytic level; in a mixture of pollen derived from
mono-trisomic plant, nulli-di pollen (n= 19+2) function as normal
wheat pollen (n=21) only when the missing chr. & extra chr. in
nulli-di pollen were homeologous each other
2. Loss of 5B Chr.; pairing b/w non-homologous chr.
3. Genetic evidence of homeology by using genes on same
chromosomes
Homology has extended to genera Triticae (Aegilops, Agropyron,
Haynaldia, Hordium, Secale etc.)
“Homeology proved the common ancestry & Genome didn't
differentiate very much after their initial divergence”
15
D. Diploidizing (5B) system in Bread wheat
• Due to presence of Ph1 gene on long arm of 5B chr.
• Diploidizing system function even in hemizygous condition
(monosomic), but hemizygous inefficient (in 6x Festuca) &
hemizygous insufficient (in polyploid Hordeum)
16
Meiotic behavior
• Monosomics form bivalents and solitary univalents, rarely
trivalents
• Univalent shift; formation of more than one univalents due to
failure of association of one/more pairs homologous chr.,
which gives rise to other monosomics
• The solitary univalent either gets included in one of the two
nuclei may lag/divide at AI consequently at AII it may divide
of lag
• Behavior of univalent at meiosis determines the frequency of
microspore with different chr. constitutions
• Eg: Wheat (2x-1) & tobacco (3x-1) – 75%
• Oat (3x-1) – 84-91%
17
Breeding behaviour of Monosomics
• Breeding behavior is studied by examining the progeny
obtained by selfing them and crossing them separately as
male and female parent with normal's. This helps to calculate
the frequency of functional deficient gamete relative to
normal.
• Deficient gametes produced in higher frequency but function
at low frequency in pollen
• Eg: fig.
18
Fig. Monosomic selfing
19
Production of monosomic series in a new variety
• Wheat; monosomic series was initially produced in the variety
Chinese Spring
• For convenient use, one may like to use monosomic series in a
popular variety of his country
Chinese Spring (monosomic) X popular variety
F1 X popular variety
BC1 X
BC2
BC4-6
20
Checking the identity of monosomics
Monosomic X nullisomic
True identity: F1 20II
Mistaken identity: 19II + 1II
Monosomic X ditelocentric
True identity: F1 20II + 1 telo
Mistaken identity: 19II + 1heteromorph
21
Uses of Monosomics
• The preparation of linkage map in polyploid species has been
difficult due to the presence of duplicate genes or due to
polygenic inheritance. So a technique known as monosomic
analysis has been successfully used.
• Based on nature of gene, different types of monosomic
analysis are possible.
22
Different types of Monosomic analysis
1. Use of monosomic analysis to locate dominant genes to
chromosomes
1. Use of monosomic analysis to locate recessive genes to
chromosomes
2. Use of monosomic analysis in locating genes to chromosomes for
digenic trait
3. Use of monosomic analysis in locating genes through intervarietal
chromosomal substitutions
4. Use of monosomic analysis in locating genes on chromosome arms
23
Locating dominant gene by absence of
expression in the nullisomic
• Nullisomic for the chr. carrying the gene should express the
effect of absence
• Self each of 21 monosomics & observe for the absence of
gene expression
• Eg: 3D- Red kernel
4B- awn suppression
24
Locate dominant genes to chromosomes through F1
analysis
25
Locating recessive genes through analysis of F2
26
Monosomic analysis of genic male-sterility in
hexaploid wheat
• In most of the crops male sterility is controlled by recessive nuclear
gene ms
• Recently a novel genie male-sterility was reported by Singh (2002)
where the male-sterility was incomplete, therefore, it was
designated as p-mst (partial genie male sterility)
• In the present study, an attempt has been made to locate ms gene
on specific chromosome of partial genie male sterile (p-mst) strain.
• Material & methods
• The partial genie male-sterility strain of T. aestivum (2n=42) from
the Department of Genetics, IARI, New Delhi (full awning, single
gene dwarf, late maturing, resistant to stem and leaf rusts of wheat.
It produces 10 to 12% selfed seeds)
• The 21 aneuploid lines of cv. Chinese Spring used were originally
produced by Sears (1954) (awnless and susceptible to rusts)
27
Dalmir Singh and P.K. Biswas (2002), Wheat Information Service, Number 95: 1-4 Research article
Contd..
28
P-mst F
15: 1
4A F1
425
(Fertile)
151
(partial sterile)
6B F1
280
(Fertile)
101
(Partial sterile)
Result
• All the 21 monosomic F1 hybrids produced less number of
seeds per spikelet than disomic F1 hybrid
• A good fit to a ratio of 15 fertile: 1 sterile was obtained in the
F2's of the disomic cross (control) as well as in the 19 families
of the monosomic F2's. In crosses involving chromosomes 4A
and 6B expected digenic segregation was not observed.
29
conclusion
• Location of gene for mst trait on chromosome 4A confirms the
finding of Driscoll (1975) and Kleijer and Fossati (1976)
where ms genes of Pugsley and Probus mutants were located,
on chromosome 4A.
• Location of another gene for mst trait on chromosome 6B, is
in support of the findings reported by Sears (1954)
30
Locating hemizygous ineffective genes through analysis of F3
31
A - ss 73
- - ss 24
A – ss 3
Use of monosomic analysis in locating genes to
chromosomes for digenic trait
32
locating genes through inter-varietal chromosomal
substitutions
• If substitution leads to major
change in the morphology for the
character under investigation,
gene(s) for these characters are
present on these chr.
• Kuspira and Unrau (1957);
identified genes for awning,
earliness, lodging, plant height,
protein content and 1000 kernel
wt.
33
Locating genes on chromosome arms
34
`
35
Monosomics in diploids
• Chr. 4- Drosophila melanogastor
• X - human being
• Datura stramonium
• Nicotiana alata
• N. longsdorfii
• Solanum lycopersicon
• Zea mays
Most of the cases, monosomic condition was not transmitted to
progeny
36
Monosomics in diploid obtained by-
• Rare monosomics in the progeny of normal diploid
• Mutation treatment
• Progeny of aneuploids, haploid and polyploids
• Interspecific cross
• Progeny of specific genetic system (Maize)
37
Monosomics in human
Monosomy X (Turner Syndrome) is a
karyotypic condition caused by non-
disjunction of X chromosomes at Meiosis
I or II. Frequency is 1 in 5000 female births
Symptoms
Short stature
Fold of skin
Shield shaped thorax
Constriction of aorta
Widely shaped nipples
Poor breast development
Elbow deformity
Rudimentary ovaries
Brown spots
Small finger nails
Monosomics for all human
autosomes die in utero
38
39

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MONOSOMICS: Understanding Meiotic Behavior and Uses in Crop Improvement

  • 1. MONOSOMICS Manjappa Ph. D Scholar Dept. of Genetics and Plant Breeding 1
  • 2. Nutshell • Introduction • Methods of production • Identification of monosomics • Meiotic behavior of monosomics • Breeding behavior • Checking identity of monosomics • Locating genes on polyploid crops • Human disorder 2
  • 3. Introduction  Polyploids can tolerate not diploid (except Maize - diploid or segmental polyploid) History:  Scattered work on monosomics initiated in early 1920’s  Systematic work started by-  Clausen & Cameron (1944)- Nicotiana tabacum  Sears (1954)- Hexaploid wheat  Endrizzi et al. (1963, 84, 85)- Gossypium hirsutum 3
  • 4. Methods of Monosomic Production 1. From haploids: 4
  • 5. 2. Backcrosses of interspecific hybrids 5
  • 6. Production of monosomics of D genome in bread wheat 6
  • 7. 3. From partially asynaptic plants 7
  • 8. 4. Irradiation treatment: Inflorescence is irradiated  Non-disjunction of normal bivalents- production of gametes with 2x+1 and 2x-1 5. Spontaneous production:  Occasional non-disjunction of bivalents during meiosis 8
  • 10. Description and identification of monosomics • Generally polyploids don’t show drastic alterations in morphology (bcz. addition of chromosome to diploid creates greater disturbance) • Chromosome count in mitosis & univalents in meiosis N. Tabaccum x N. sylverstris (Monosomic) 12I (T) + 11I (S) 12I (S) 12I (T) + 11II + 1I (S) 2n= 11II + 13I Monosomic for Sylverstris genome N. Tabaccum x N. sylverstris (Monosomic) 11I (T) + 12I (S) 12I (S) 11I (T) + 12II + 1I (S) 2n= 12II + 11I Not monosomic for Sylverstris genome 10
  • 11. Identification in Cotton 1. Cross monosomic with proginator 2. Study of meiosis in hybrids b/w monosomic and translocated testers that are available in cotton 11
  • 12. 12
  • 13. Identification of monosomics & related phenomena in wheat a) Classification of monosomics in durum group (A & B genome) & D genome T. aestivum X T. durum 2n= 20II+1I (ABD) 2n= 14II (AB) Meiosis: 14II + 6I (AB) (D) Monosomic in D genome T. aestivum X T. durum 2n= 20II+1I (ABD) 2n= 14II (AB) Meiosis: 13II + 8I (AB) (D+A/B) Monosomic in A/B genome 13
  • 14. Distinction b/w monosomics of A & B genomes • If telocentric chr. was involved in the formation of heteromorphic bivalent, then the monosomic belongs to A genome if not to B genome Monosomics X Amphidiploid (Ditelocentric) AADD (2n= 20II + 1II telo) (2n= 14II) F1: 34 + telo 14
  • 15. C. Assessing chr. to homeologous groups in Wheat (compensating nullisomic-tetrasomic lines • It has been demo. that the presence of an extra chr. may separately compensate phenotypically for the absence of each of the other two chr. “This is an evidence of homeology among 3 chr.” • Confirmed by- 1. Study at gametophytic level; in a mixture of pollen derived from mono-trisomic plant, nulli-di pollen (n= 19+2) function as normal wheat pollen (n=21) only when the missing chr. & extra chr. in nulli-di pollen were homeologous each other 2. Loss of 5B Chr.; pairing b/w non-homologous chr. 3. Genetic evidence of homeology by using genes on same chromosomes Homology has extended to genera Triticae (Aegilops, Agropyron, Haynaldia, Hordium, Secale etc.) “Homeology proved the common ancestry & Genome didn't differentiate very much after their initial divergence” 15
  • 16. D. Diploidizing (5B) system in Bread wheat • Due to presence of Ph1 gene on long arm of 5B chr. • Diploidizing system function even in hemizygous condition (monosomic), but hemizygous inefficient (in 6x Festuca) & hemizygous insufficient (in polyploid Hordeum) 16
  • 17. Meiotic behavior • Monosomics form bivalents and solitary univalents, rarely trivalents • Univalent shift; formation of more than one univalents due to failure of association of one/more pairs homologous chr., which gives rise to other monosomics • The solitary univalent either gets included in one of the two nuclei may lag/divide at AI consequently at AII it may divide of lag • Behavior of univalent at meiosis determines the frequency of microspore with different chr. constitutions • Eg: Wheat (2x-1) & tobacco (3x-1) – 75% • Oat (3x-1) – 84-91% 17
  • 18. Breeding behaviour of Monosomics • Breeding behavior is studied by examining the progeny obtained by selfing them and crossing them separately as male and female parent with normal's. This helps to calculate the frequency of functional deficient gamete relative to normal. • Deficient gametes produced in higher frequency but function at low frequency in pollen • Eg: fig. 18
  • 20. Production of monosomic series in a new variety • Wheat; monosomic series was initially produced in the variety Chinese Spring • For convenient use, one may like to use monosomic series in a popular variety of his country Chinese Spring (monosomic) X popular variety F1 X popular variety BC1 X BC2 BC4-6 20
  • 21. Checking the identity of monosomics Monosomic X nullisomic True identity: F1 20II Mistaken identity: 19II + 1II Monosomic X ditelocentric True identity: F1 20II + 1 telo Mistaken identity: 19II + 1heteromorph 21
  • 22. Uses of Monosomics • The preparation of linkage map in polyploid species has been difficult due to the presence of duplicate genes or due to polygenic inheritance. So a technique known as monosomic analysis has been successfully used. • Based on nature of gene, different types of monosomic analysis are possible. 22
  • 23. Different types of Monosomic analysis 1. Use of monosomic analysis to locate dominant genes to chromosomes 1. Use of monosomic analysis to locate recessive genes to chromosomes 2. Use of monosomic analysis in locating genes to chromosomes for digenic trait 3. Use of monosomic analysis in locating genes through intervarietal chromosomal substitutions 4. Use of monosomic analysis in locating genes on chromosome arms 23
  • 24. Locating dominant gene by absence of expression in the nullisomic • Nullisomic for the chr. carrying the gene should express the effect of absence • Self each of 21 monosomics & observe for the absence of gene expression • Eg: 3D- Red kernel 4B- awn suppression 24
  • 25. Locate dominant genes to chromosomes through F1 analysis 25
  • 26. Locating recessive genes through analysis of F2 26
  • 27. Monosomic analysis of genic male-sterility in hexaploid wheat • In most of the crops male sterility is controlled by recessive nuclear gene ms • Recently a novel genie male-sterility was reported by Singh (2002) where the male-sterility was incomplete, therefore, it was designated as p-mst (partial genie male sterility) • In the present study, an attempt has been made to locate ms gene on specific chromosome of partial genie male sterile (p-mst) strain. • Material & methods • The partial genie male-sterility strain of T. aestivum (2n=42) from the Department of Genetics, IARI, New Delhi (full awning, single gene dwarf, late maturing, resistant to stem and leaf rusts of wheat. It produces 10 to 12% selfed seeds) • The 21 aneuploid lines of cv. Chinese Spring used were originally produced by Sears (1954) (awnless and susceptible to rusts) 27 Dalmir Singh and P.K. Biswas (2002), Wheat Information Service, Number 95: 1-4 Research article
  • 28. Contd.. 28 P-mst F 15: 1 4A F1 425 (Fertile) 151 (partial sterile) 6B F1 280 (Fertile) 101 (Partial sterile)
  • 29. Result • All the 21 monosomic F1 hybrids produced less number of seeds per spikelet than disomic F1 hybrid • A good fit to a ratio of 15 fertile: 1 sterile was obtained in the F2's of the disomic cross (control) as well as in the 19 families of the monosomic F2's. In crosses involving chromosomes 4A and 6B expected digenic segregation was not observed. 29
  • 30. conclusion • Location of gene for mst trait on chromosome 4A confirms the finding of Driscoll (1975) and Kleijer and Fossati (1976) where ms genes of Pugsley and Probus mutants were located, on chromosome 4A. • Location of another gene for mst trait on chromosome 6B, is in support of the findings reported by Sears (1954) 30
  • 31. Locating hemizygous ineffective genes through analysis of F3 31 A - ss 73 - - ss 24 A – ss 3
  • 32. Use of monosomic analysis in locating genes to chromosomes for digenic trait 32
  • 33. locating genes through inter-varietal chromosomal substitutions • If substitution leads to major change in the morphology for the character under investigation, gene(s) for these characters are present on these chr. • Kuspira and Unrau (1957); identified genes for awning, earliness, lodging, plant height, protein content and 1000 kernel wt. 33
  • 34. Locating genes on chromosome arms 34
  • 35. ` 35
  • 36. Monosomics in diploids • Chr. 4- Drosophila melanogastor • X - human being • Datura stramonium • Nicotiana alata • N. longsdorfii • Solanum lycopersicon • Zea mays Most of the cases, monosomic condition was not transmitted to progeny 36
  • 37. Monosomics in diploid obtained by- • Rare monosomics in the progeny of normal diploid • Mutation treatment • Progeny of aneuploids, haploid and polyploids • Interspecific cross • Progeny of specific genetic system (Maize) 37
  • 38. Monosomics in human Monosomy X (Turner Syndrome) is a karyotypic condition caused by non- disjunction of X chromosomes at Meiosis I or II. Frequency is 1 in 5000 female births Symptoms Short stature Fold of skin Shield shaped thorax Constriction of aorta Widely shaped nipples Poor breast development Elbow deformity Rudimentary ovaries Brown spots Small finger nails Monosomics for all human autosomes die in utero 38
  • 39. 39