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Rapid Improvement
of
Quality Protein
Maize
Vinod Kumar Goyal & Lokesh Gour
DEPARTMENT OF PLANT BREEDING & GENETICSDEPARTMENT OF PLANT BREEDING & GENETICS
JAWAHARLAL NEHRU KRISHI VISHWA VIDYALAYAJAWAHARLAL NEHRU KRISHI VISHWA VIDYALAYA
JABALPUR , MADHYA PRADESH (INDIA)JABALPUR , MADHYA PRADESH (INDIA)
• Name of Crop: Maize
• Botanical Name: Zea mays L.
• Family: Poaceae
• Chromosome No.: 2n = 20
• Center of Origin: Central America (Mexico)
• Mode of pollination : Cross pollination
• Out crossing percentage: 95%
•India is the sixth largest producer of maize in the world contributing 2%
of the global production.
•In India maize is the third most important crop after rice and wheat
and accounts for 9% of the total food grain production in country.
•The important maize growing states are Karnataka, Andhra Pradesh,
Bihar, Punjab and Himachal Pradesh
Maize ?
• Primarily animal feed in
– East and Southeast Asia
• Primarily human food in
– Africa
– Central America
– South Asia
• 15-56% of total daily calories
– For several hundred million
people
– Including weaning children
– in 25 countries
Maize in developing countries
Nutritional
Limitations of Maize
Low availability of
niacin
Low lysine and
tryptophan
Need to improve
nutrition
– Essential amino acids
– Major seed protein (60%)
– Lysine level about 2%
• 4% recommended by
FAO
– For humans and animals
– Quality Protein Maize:
QPM
Quality Protein Maize
It is an improved variety of
maize which contains higher
amount of lysine and
tryptophan with lower amount
of leucine and isoleucine in
the endosperm than those
contained in normal maize.
Need Of Quality Protein Maize
 However, malnutrition still remains a widespread problem, and is
particularly severe in developing countries with low per capita income.
 Animal protein, of course being of higher quality, is scarce and expensive,
thereby unavailable to a vast sector of the population.
 Maize is a major cereal crop and plays very important role in human and
animal nutrition in a number of developed and developing countries
worldwide, derive their protein and calorie requirements from maize.
 With its high content of carbohydrates, fats, proteins, some of the important
vitamins and minerals, maize acquired a well-deserved reputation as a “poor
man’s nutria-cereal”.
 Normal maize varieties are deficient to two essential amino acids, lysine and
tryptophan (Azevedo and Arruda, 2010; Mbuya et al., 2011).
Discovery of QPM
?
T. B. Osborne
Proteins of the
maize kernel
deficient in
lysine and
tryptophan
(1914)
L. B. Mendel
Reported second mutant for change in
amino acid composition (1935) - fl2
Discovered first high lysine mutant
(1935) - o2
Mumm
Dr. Howard
Jones
Researchers at
Purdue University
rediscovered a
recessive mutant
called Opaque-2
(1964)
Edwin
Theodore
Mertz
Oliver E.
Nelson
Nutritional superiority of QPM: A
compression
Lysine content
QPM protein
contains
Biological
Value
Normal -1.3 g per 100 g endosperm protein.
 o2-3.3 to 4.0 g per 100 g of endosperm protein.
55% more tryptophan.
30% more lysine.
38% less leucine than that of normal maize.
Normal maize protein is 45%.
o2 maize is 80%..
Protein intake
utilization
Maize required
for nitrogen
equilibrium/ kg
of body wt.
Other
nutritional
benefits
Common maize - 37%
o2 maize protein -74%
Normal maize -24 g
QPM-8 g.
Higher tryptophan and lower leucine content,
Higher calcium and carbohydrate and carotene
Higher niacin
Cont.
Wild-type
Opaque 2
Opaque2 – a gene for improving quality of protein
in maize
 A natural spontaneous maize mutant with soft and opaque grain was
found in a maize fields in USA during the 1920s which was later
named as opaque2 (o2) maize by Singleton.
 The mutant was passed onto Mertz at Purdue University, USA, who,
in turn, reported that the o2 homozygous maize contained
substantially higher lysine (+69%) in the grain endosperm compared
to normal maize.
 The increase in lysine content doubled the biological value of the o2
maize protein and this increase in protein quality is due to increase in
the ratio of non-zein to zein proteins.
Early efforts and experiences in using o2 cultivars
Initiation of Breeding programmes to develop inbred lines by using
Populations with Endosperm quality mutants, mainly o2
o2 utilization in breeding programmes
Resulted
Soft endosperm
Damaged kernels
Susceptibility to
pests and fungal
diseases
Inferior food processing
Reduced yields
Early efforts and experiences in using o2 cultivars
pleiotropic effects of this gene
Development of QPM involves three genetic systems, viz.
I. Recessive homozygous allele of the o2 gene
II. Modifiers for kernel hardness
III. Amino acid modifiers
Genetic systems and their role in enhancing the level of
limiting amino acids in QPM
• The o2 allele in homozygous condition reduces production of alpha-
zeins and triggers increase in the level of lysine and tryptophan.
• Involved in the synthesis of the
enzyme that is associated with
free lysine degradation.
Reduction in this enzyme leads
to a corresponding increase in
free lysine in the endosperm.
(I ) Recessive homozygous allele of the o2 gene
Reduced protein body size in o2
WT
o2
) Modifiers for kernel hardness
• Increased level of the gamma-zein is likely to contribute to the
recovery of hard endosperm.
• o2 modified (QPM) grains have double the amount of gamma-zeins
in the endosperm compared to the o2 mutants
• Two genes responsible for the grain hardness, mapped to the long
arm of chromosome 7 and one of them is located near the gamma-
zein gene ‘gzr 1’.
• Increased amount of granule-bound starch synthase I reflected in the
form of a change in starch structure, which was manifested as shorter
amylopectin branches and increased swelling of starch granules.
CM105 wild-type CM105 opaque 2
Starch
Starch
PB
) Amino acid modifiers
• It also affect the relative level of lysine and tryptophan content in
grain endosperm.
• Three genes associated with lysine level have been mapped to
locations on chromosome 2, 4 and 7.
• Apart from this, several major o2 modifier-QTLs located on
chromosomes 1, 7 and 9, have been recently mapped
A.Gradual increase in proportion of
soft to hard endosperms
Tucson (USA) Lopes and Larkin (1991)
B. Spatial expression of opaque-2
under different genetic backgrounds
 Conventional Breeding
 Marker Assisted Selection
Development of QPM genotypes through :
Rapid Improvement of QPM genotypes through
conventional breeding
Development of Acceptable QPM lines by combining the nutritional
advantages of o2 mutation with the o2 modifiers.
During the 1980s, CIMMYT took initiatives to convert a number of
non-QPM genotypes to QPM genotypes, they followed a ‘modified
backcrossing-cum-recurrent selection’.
During the conversion process, they also emphasized grain yield,
kernel modification, reduced ear rot incidence and other agronomic
traits.
In a short span of 5–6 years, CIMMYT could convert many normal
germplasm into QPM, which were as good as their non-QPM
counterparts for grain yield and other agronomic traits.
Two scientists of CIMMYT, Mexico, Dr. S. K. Vasal and Dr.
Evangelina Villegas, for a period of three decades led to development
of Quality Protein Maize (QPM) with hard kernel, good taste and other
consumer favouring characteristics.
QPM research and development spread from Mexico to Central and
South America, Africa, Europe and Asia.
Awarded 2000 “World food prize” for path breaking research.
Cont.
 1970 -India is one of the first few countries to focus on o2 maize and
released three o2 composites, namely Shakti, Rattan and Protina .
 1997- modified superior o2 composite ‘Shakti 1’.
 Later, India released eight QPM hybrids, seven of which were
developed from the QPM inbreds of CIMMYT as parental lines and
are of full season maturity.(HQPM1, HQPM5 ,HQPM 7 Vivek QPM
9)
 Breakthrough achievement in development of “Miracle Maize”
Cont.
Development of QPM hybrid through marker assisted
selection
In order to shorten the period required for development of QPM
hybrids through the conventional method of backcrossing, marker-
assisted selection (MAS) can be used.
Few molecular markers were already known within the o2 gene and
these makers were capable of detecting the o2 gene even in hetero-
zygous state.
To convert normal maize hybrid into QPM hybrid, a promising hybrid,
viz. Vivek Maize Hybrid 9 (developed by Vivekananda Parvatiya Krishi
Anusandhan Sansthan, Almora) was selected for converting into QPM.
CML 180
VQL 1
CML 170CM 212
VQL 2
CM 145
Recipient
o2gene
Recipients
o2gene
Donor
inbred line
Donor
inbred line
Vivek QPM 9
(92.0%) (94.44%)
o2o2 quantitative
determination
Using SSR markers
phi57, phi112,
umc 1066
Determining lysine, tryptophan , protein
Biochemical analysis
Conversion of normal maize inbreds into QPM
Gupta et al. 2009.
The hybrid was released for commercial cultivation in Himalayan
states, Andhra Pradesh, Tamil Nadu, Karnataka and Maharashtra by
the Central Seed Sub-committee on Crop Standard and Notification
in the year 2000.
Cont….
Name Pedigree Year of
Release
Maturity
group
Centre’s
name
Shakti Composite 1970 Full season AICRP
Rattan Composite 1970 Full season AICRP
Protina Composite 1970 Full season AICRP
Shakti 1 Composite 1997 Full season DMR
Shaktiman 1 (CML 142 x CML 150) xCML 186 2001 Full season Dholi
Shaktiman 2 CML 176 x CML 186 2004 Full season Dholi
HQPM 1 HKI 193-1 x HKI 163 2005 Full season Uchani
Shaktiman 3 CML 161 x CML 163 2006 Full season Dholi
Shaktiman 4 CML 161 x CML 169 2006 Full season Dholi
HQPM 5 HKI 163 x HKI 161 2007 Full season Uchani
HQPM 7 HKI 193-1 x HKI 161 2008 Full season Uchani
Vivek QPM 9 VQL 1 x VQL 2 2008 Extra early Almora
QPM cultivars released for commercial cultivation
in India
(Dhillon and Prasanna, 2001 and Gupta et al. 2009
Name Protein content in
grain (%)
Tryptophan content in
protein (%)
Shaktiman1 9.6 1.01
Shaktiman 2 9.3 1.04
HQPM 1 9.36 0.94
Shaktiman 3 9.63 0.73
Shaktiman 4 9.98 0.93
HQPM 5 9.80 0.76
HQPM 7 9.4 0.72
Vivek QPM 9 8.46 0.83
Protein content in grain (%) and Tryptophan content
in protein (%)
Recently developed QPM varieties
 Following this approach, we have now developed another elite QPM
hybrid-FQH 38
 The QPM version of Vivek Maize Hybrid 21.
 The new QPM hybrid contains more than 72% increase in tryptophan
over Vivek Maize Hybrid 21
 Anand QPM 1- developed from Anand agriculture university.
 For a country like India, with diverse agroclimatic and soil situations,
need to develop a number of QPM hybrids of different maturity
groups, viz. early, medium and late (full season).
 However, the major constraints in adoption of the QPM hybrids in
these areas are the non-availability of hybrid seeds and lack of
incentives like premium price for the QPM over normal maize grains.
 There is also a need to create awareness among the consumers and
industry for its use in food and feed.
 We are developing a linkage between the seed producers, farmers and
the industry to bring about the much needed synergy in development
and utilization of QPM that will help in reducing protein
Conclusion
Anonymous 2008. The Gazette of India, Extraordinary. Part II. Section
3, subsection (ii), No. 1448, Serial No. 28, Ministry of Agriculture,
Department of Agriculture and Cooperation, New Delhi, 16
October.
Babu R. 2005. Two generation marker-aided backcrossing for rapid
conversion of normal maize lines to quality protein maize (QPM).
Theor. Appl. Genet. (111): 888–897.
Sharma R, Maloo S R and Joshi A. 2014. Genetic variability analysis in
diverse maize genotypes. Journal of plant breeding 5(3):545-551.
Some of Referances
Vasal SK, Villegas E, Bjarnason M, Gelaw B, and Goertz P. 1980.
Genetic Modifiers and Breeding Strategies in Developing Hard
Endosperm opaque-2 Materials, pp. 37–73 in Improvement of
Quality Traits of Maize for Grain and Silage Use, edited by
Pollmer WG and Phipps RH. Nijhoff M, London.
Yadav VK. 2001. Quality protein maize. In Production Technology and
Value Addition, Directorate of Maize Research, New Delhi, p. 4.
Cont…

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Rapid improvement of quality protein maize

  • 1. Rapid Improvement of Quality Protein Maize Vinod Kumar Goyal & Lokesh Gour DEPARTMENT OF PLANT BREEDING & GENETICSDEPARTMENT OF PLANT BREEDING & GENETICS JAWAHARLAL NEHRU KRISHI VISHWA VIDYALAYAJAWAHARLAL NEHRU KRISHI VISHWA VIDYALAYA JABALPUR , MADHYA PRADESH (INDIA)JABALPUR , MADHYA PRADESH (INDIA)
  • 2. • Name of Crop: Maize • Botanical Name: Zea mays L. • Family: Poaceae • Chromosome No.: 2n = 20 • Center of Origin: Central America (Mexico) • Mode of pollination : Cross pollination • Out crossing percentage: 95% •India is the sixth largest producer of maize in the world contributing 2% of the global production. •In India maize is the third most important crop after rice and wheat and accounts for 9% of the total food grain production in country. •The important maize growing states are Karnataka, Andhra Pradesh, Bihar, Punjab and Himachal Pradesh Maize ?
  • 3. • Primarily animal feed in – East and Southeast Asia • Primarily human food in – Africa – Central America – South Asia • 15-56% of total daily calories – For several hundred million people – Including weaning children – in 25 countries Maize in developing countries
  • 4. Nutritional Limitations of Maize Low availability of niacin Low lysine and tryptophan Need to improve nutrition – Essential amino acids – Major seed protein (60%) – Lysine level about 2% • 4% recommended by FAO – For humans and animals – Quality Protein Maize: QPM
  • 5. Quality Protein Maize It is an improved variety of maize which contains higher amount of lysine and tryptophan with lower amount of leucine and isoleucine in the endosperm than those contained in normal maize.
  • 6. Need Of Quality Protein Maize  However, malnutrition still remains a widespread problem, and is particularly severe in developing countries with low per capita income.  Animal protein, of course being of higher quality, is scarce and expensive, thereby unavailable to a vast sector of the population.  Maize is a major cereal crop and plays very important role in human and animal nutrition in a number of developed and developing countries worldwide, derive their protein and calorie requirements from maize.  With its high content of carbohydrates, fats, proteins, some of the important vitamins and minerals, maize acquired a well-deserved reputation as a “poor man’s nutria-cereal”.  Normal maize varieties are deficient to two essential amino acids, lysine and tryptophan (Azevedo and Arruda, 2010; Mbuya et al., 2011).
  • 8. T. B. Osborne Proteins of the maize kernel deficient in lysine and tryptophan (1914) L. B. Mendel
  • 9. Reported second mutant for change in amino acid composition (1935) - fl2 Discovered first high lysine mutant (1935) - o2 Mumm Dr. Howard Jones
  • 10. Researchers at Purdue University rediscovered a recessive mutant called Opaque-2 (1964) Edwin Theodore Mertz Oliver E. Nelson
  • 11. Nutritional superiority of QPM: A compression Lysine content QPM protein contains Biological Value Normal -1.3 g per 100 g endosperm protein.  o2-3.3 to 4.0 g per 100 g of endosperm protein. 55% more tryptophan. 30% more lysine. 38% less leucine than that of normal maize. Normal maize protein is 45%. o2 maize is 80%..
  • 12. Protein intake utilization Maize required for nitrogen equilibrium/ kg of body wt. Other nutritional benefits Common maize - 37% o2 maize protein -74% Normal maize -24 g QPM-8 g. Higher tryptophan and lower leucine content, Higher calcium and carbohydrate and carotene Higher niacin Cont.
  • 14. Opaque2 – a gene for improving quality of protein in maize  A natural spontaneous maize mutant with soft and opaque grain was found in a maize fields in USA during the 1920s which was later named as opaque2 (o2) maize by Singleton.  The mutant was passed onto Mertz at Purdue University, USA, who, in turn, reported that the o2 homozygous maize contained substantially higher lysine (+69%) in the grain endosperm compared to normal maize.  The increase in lysine content doubled the biological value of the o2 maize protein and this increase in protein quality is due to increase in the ratio of non-zein to zein proteins.
  • 15. Early efforts and experiences in using o2 cultivars Initiation of Breeding programmes to develop inbred lines by using Populations with Endosperm quality mutants, mainly o2
  • 16. o2 utilization in breeding programmes Resulted Soft endosperm Damaged kernels Susceptibility to pests and fungal diseases Inferior food processing Reduced yields Early efforts and experiences in using o2 cultivars pleiotropic effects of this gene
  • 17. Development of QPM involves three genetic systems, viz. I. Recessive homozygous allele of the o2 gene II. Modifiers for kernel hardness III. Amino acid modifiers Genetic systems and their role in enhancing the level of limiting amino acids in QPM
  • 18. • The o2 allele in homozygous condition reduces production of alpha- zeins and triggers increase in the level of lysine and tryptophan. • Involved in the synthesis of the enzyme that is associated with free lysine degradation. Reduction in this enzyme leads to a corresponding increase in free lysine in the endosperm. (I ) Recessive homozygous allele of the o2 gene Reduced protein body size in o2 WT o2
  • 19. ) Modifiers for kernel hardness • Increased level of the gamma-zein is likely to contribute to the recovery of hard endosperm. • o2 modified (QPM) grains have double the amount of gamma-zeins in the endosperm compared to the o2 mutants • Two genes responsible for the grain hardness, mapped to the long arm of chromosome 7 and one of them is located near the gamma- zein gene ‘gzr 1’.
  • 20. • Increased amount of granule-bound starch synthase I reflected in the form of a change in starch structure, which was manifested as shorter amylopectin branches and increased swelling of starch granules. CM105 wild-type CM105 opaque 2 Starch Starch PB
  • 21. ) Amino acid modifiers • It also affect the relative level of lysine and tryptophan content in grain endosperm. • Three genes associated with lysine level have been mapped to locations on chromosome 2, 4 and 7. • Apart from this, several major o2 modifier-QTLs located on chromosomes 1, 7 and 9, have been recently mapped
  • 22. A.Gradual increase in proportion of soft to hard endosperms Tucson (USA) Lopes and Larkin (1991) B. Spatial expression of opaque-2 under different genetic backgrounds
  • 23.  Conventional Breeding  Marker Assisted Selection Development of QPM genotypes through :
  • 24. Rapid Improvement of QPM genotypes through conventional breeding Development of Acceptable QPM lines by combining the nutritional advantages of o2 mutation with the o2 modifiers. During the 1980s, CIMMYT took initiatives to convert a number of non-QPM genotypes to QPM genotypes, they followed a ‘modified backcrossing-cum-recurrent selection’. During the conversion process, they also emphasized grain yield, kernel modification, reduced ear rot incidence and other agronomic traits. In a short span of 5–6 years, CIMMYT could convert many normal germplasm into QPM, which were as good as their non-QPM counterparts for grain yield and other agronomic traits.
  • 25. Two scientists of CIMMYT, Mexico, Dr. S. K. Vasal and Dr. Evangelina Villegas, for a period of three decades led to development of Quality Protein Maize (QPM) with hard kernel, good taste and other consumer favouring characteristics. QPM research and development spread from Mexico to Central and South America, Africa, Europe and Asia. Awarded 2000 “World food prize” for path breaking research. Cont.
  • 26.  1970 -India is one of the first few countries to focus on o2 maize and released three o2 composites, namely Shakti, Rattan and Protina .  1997- modified superior o2 composite ‘Shakti 1’.  Later, India released eight QPM hybrids, seven of which were developed from the QPM inbreds of CIMMYT as parental lines and are of full season maturity.(HQPM1, HQPM5 ,HQPM 7 Vivek QPM 9)  Breakthrough achievement in development of “Miracle Maize” Cont.
  • 27. Development of QPM hybrid through marker assisted selection In order to shorten the period required for development of QPM hybrids through the conventional method of backcrossing, marker- assisted selection (MAS) can be used. Few molecular markers were already known within the o2 gene and these makers were capable of detecting the o2 gene even in hetero- zygous state. To convert normal maize hybrid into QPM hybrid, a promising hybrid, viz. Vivek Maize Hybrid 9 (developed by Vivekananda Parvatiya Krishi Anusandhan Sansthan, Almora) was selected for converting into QPM.
  • 28. CML 180 VQL 1 CML 170CM 212 VQL 2 CM 145 Recipient o2gene Recipients o2gene Donor inbred line Donor inbred line Vivek QPM 9 (92.0%) (94.44%) o2o2 quantitative determination Using SSR markers phi57, phi112, umc 1066 Determining lysine, tryptophan , protein Biochemical analysis Conversion of normal maize inbreds into QPM Gupta et al. 2009.
  • 29. The hybrid was released for commercial cultivation in Himalayan states, Andhra Pradesh, Tamil Nadu, Karnataka and Maharashtra by the Central Seed Sub-committee on Crop Standard and Notification in the year 2000. Cont….
  • 30. Name Pedigree Year of Release Maturity group Centre’s name Shakti Composite 1970 Full season AICRP Rattan Composite 1970 Full season AICRP Protina Composite 1970 Full season AICRP Shakti 1 Composite 1997 Full season DMR Shaktiman 1 (CML 142 x CML 150) xCML 186 2001 Full season Dholi Shaktiman 2 CML 176 x CML 186 2004 Full season Dholi HQPM 1 HKI 193-1 x HKI 163 2005 Full season Uchani Shaktiman 3 CML 161 x CML 163 2006 Full season Dholi Shaktiman 4 CML 161 x CML 169 2006 Full season Dholi HQPM 5 HKI 163 x HKI 161 2007 Full season Uchani HQPM 7 HKI 193-1 x HKI 161 2008 Full season Uchani Vivek QPM 9 VQL 1 x VQL 2 2008 Extra early Almora QPM cultivars released for commercial cultivation in India (Dhillon and Prasanna, 2001 and Gupta et al. 2009
  • 31. Name Protein content in grain (%) Tryptophan content in protein (%) Shaktiman1 9.6 1.01 Shaktiman 2 9.3 1.04 HQPM 1 9.36 0.94 Shaktiman 3 9.63 0.73 Shaktiman 4 9.98 0.93 HQPM 5 9.80 0.76 HQPM 7 9.4 0.72 Vivek QPM 9 8.46 0.83 Protein content in grain (%) and Tryptophan content in protein (%)
  • 32. Recently developed QPM varieties  Following this approach, we have now developed another elite QPM hybrid-FQH 38  The QPM version of Vivek Maize Hybrid 21.  The new QPM hybrid contains more than 72% increase in tryptophan over Vivek Maize Hybrid 21  Anand QPM 1- developed from Anand agriculture university.
  • 33.  For a country like India, with diverse agroclimatic and soil situations, need to develop a number of QPM hybrids of different maturity groups, viz. early, medium and late (full season).  However, the major constraints in adoption of the QPM hybrids in these areas are the non-availability of hybrid seeds and lack of incentives like premium price for the QPM over normal maize grains.  There is also a need to create awareness among the consumers and industry for its use in food and feed.  We are developing a linkage between the seed producers, farmers and the industry to bring about the much needed synergy in development and utilization of QPM that will help in reducing protein Conclusion
  • 34. Anonymous 2008. The Gazette of India, Extraordinary. Part II. Section 3, subsection (ii), No. 1448, Serial No. 28, Ministry of Agriculture, Department of Agriculture and Cooperation, New Delhi, 16 October. Babu R. 2005. Two generation marker-aided backcrossing for rapid conversion of normal maize lines to quality protein maize (QPM). Theor. Appl. Genet. (111): 888–897. Sharma R, Maloo S R and Joshi A. 2014. Genetic variability analysis in diverse maize genotypes. Journal of plant breeding 5(3):545-551. Some of Referances
  • 35. Vasal SK, Villegas E, Bjarnason M, Gelaw B, and Goertz P. 1980. Genetic Modifiers and Breeding Strategies in Developing Hard Endosperm opaque-2 Materials, pp. 37–73 in Improvement of Quality Traits of Maize for Grain and Silage Use, edited by Pollmer WG and Phipps RH. Nijhoff M, London. Yadav VK. 2001. Quality protein maize. In Production Technology and Value Addition, Directorate of Maize Research, New Delhi, p. 4. Cont…