This document summarizes a seminar on doubled haploids (DH). It defines a DH as an individual with a doubled set of chromosomes from a haploid cell. It discusses the history of DH development, including early work in the 1920s. It also covers methods for producing haploids, identifying haploids, doubling chromosomes, and applications of DHs in plant breeding like QTL mapping, backcrossing, hybrid sorting, and cultivar development. DHs allow fixing of traits in one or two generations, faster development of pure lines and cultivars compared to conventional methods.
Marker Assisted Selection in Crop BreedingPawan Chauhan
Marker Assisted Selection is a value addition to conventional methods of Crop Breeding. It has been gaining importance in plant breeding with new generation of plant breeders and to get accurate and fast desired result from plant breeding.
Multiple inbred founder lines are inter-mated for several generations prior to creating inbred lines, resulting in a diverse population whose genomes are fine scale mosaics of contributions from all founders.
SELECTION METHODS IN SELF-POLLINATED CROPS viz., mass selection, pureline sel...AMIT RANA Ph. D Scholar
MASS SELECTION
Mass selection is a method of breeding in which individual plants are selected on the basis of phenotype from a mixed population , their seeds are bulked and used to grow the next generation.
Selection cycle may be repeated one or more times to increase the frequency of favorable alleles - phenotypic recurrent selection.
PURELINE SELECTION
A pureline is the progeny of a single homozygous plant of a self-pollinated species. All the plants in a pureline have the same genotype and the phenotypic variation within a pureline is due to the environment alone and has no genetic basis. However, variation within a pureline is not heritable. Hence selection in a pureline is not effective. Johannsen (1903,1926), a Danish biologist, developed the concept of pureline theory working with Princess variety of French bean (Phaseolus vulgaris), which showed variation for seed size. From a commercial seed lot he selected seeds of different sizes and grew them separately. The progenies differed in seed size. Progenies from larger seeds produced larger seeds than those obtained from smaller seeds. This clearly showed that the variation in seed size in the commercial seed lot of princess variety had a genetic base. As a result selection for seed size was effective.
Introduction
PEDIGREE SELECTION
Pedigree selection is a widely used method of breeding self-pollinated species.
A key difference between pedigree selection and mass selection or pure-line selection is that hybridization is used to generate variability (for the base population), unlike the other methods in which production of genetic variation is not a feature.
The method was first described by H. H. Lowe in 1927.
Pedigree selection is a breeding method in which the breeder keeps records of the ancestry of the cultivar.
The base population is established by crossing selected parents, followed by handling an actively segregating population.
Documentation of the pedigree enables breeders to trace parent–progeny back to an individual F2 plant from any subsequent generation.
The breeder should develop an effective, easy to maintain system of record keeping.
Pedigree selection is applicable to breeding species that allow individual plants to be observed, described, and harvested separately.
Mutagenesis is the process by which the genetic information
of an organism is changed in a stable manner.
The term ‘mutation breeding’ has become popular as it
draws attention to deliberate efforts of breeders and
the specific techniques they have used in creating and
harnessing desired variation in developing elite breeding
lines and cultivated varieties.
Definitions, variety production release and notification in india and pakistsudha2555
Maintenance breeding definition of variety, cultivar, EDS, IDV, reference variety. Variety development, notification and release procedure in India and Pakistan
Within the last twenty years, molecular biology has revolutionized conventional breeding techniques in all areas. Biochemical and Molecular techniques have shortened the duration of breeding programs from years to months, weeks, or eliminated the need for them all together. The use of molecular markers in conventional breeding techniques has also improved the accuracy of crosses and allowed breeders to produce strains with combined traits that were impossible before the advent of DNA technology
Marker Assisted Selection in Crop BreedingPawan Chauhan
Marker Assisted Selection is a value addition to conventional methods of Crop Breeding. It has been gaining importance in plant breeding with new generation of plant breeders and to get accurate and fast desired result from plant breeding.
Multiple inbred founder lines are inter-mated for several generations prior to creating inbred lines, resulting in a diverse population whose genomes are fine scale mosaics of contributions from all founders.
SELECTION METHODS IN SELF-POLLINATED CROPS viz., mass selection, pureline sel...AMIT RANA Ph. D Scholar
MASS SELECTION
Mass selection is a method of breeding in which individual plants are selected on the basis of phenotype from a mixed population , their seeds are bulked and used to grow the next generation.
Selection cycle may be repeated one or more times to increase the frequency of favorable alleles - phenotypic recurrent selection.
PURELINE SELECTION
A pureline is the progeny of a single homozygous plant of a self-pollinated species. All the plants in a pureline have the same genotype and the phenotypic variation within a pureline is due to the environment alone and has no genetic basis. However, variation within a pureline is not heritable. Hence selection in a pureline is not effective. Johannsen (1903,1926), a Danish biologist, developed the concept of pureline theory working with Princess variety of French bean (Phaseolus vulgaris), which showed variation for seed size. From a commercial seed lot he selected seeds of different sizes and grew them separately. The progenies differed in seed size. Progenies from larger seeds produced larger seeds than those obtained from smaller seeds. This clearly showed that the variation in seed size in the commercial seed lot of princess variety had a genetic base. As a result selection for seed size was effective.
Introduction
PEDIGREE SELECTION
Pedigree selection is a widely used method of breeding self-pollinated species.
A key difference between pedigree selection and mass selection or pure-line selection is that hybridization is used to generate variability (for the base population), unlike the other methods in which production of genetic variation is not a feature.
The method was first described by H. H. Lowe in 1927.
Pedigree selection is a breeding method in which the breeder keeps records of the ancestry of the cultivar.
The base population is established by crossing selected parents, followed by handling an actively segregating population.
Documentation of the pedigree enables breeders to trace parent–progeny back to an individual F2 plant from any subsequent generation.
The breeder should develop an effective, easy to maintain system of record keeping.
Pedigree selection is applicable to breeding species that allow individual plants to be observed, described, and harvested separately.
Mutagenesis is the process by which the genetic information
of an organism is changed in a stable manner.
The term ‘mutation breeding’ has become popular as it
draws attention to deliberate efforts of breeders and
the specific techniques they have used in creating and
harnessing desired variation in developing elite breeding
lines and cultivated varieties.
Definitions, variety production release and notification in india and pakistsudha2555
Maintenance breeding definition of variety, cultivar, EDS, IDV, reference variety. Variety development, notification and release procedure in India and Pakistan
Within the last twenty years, molecular biology has revolutionized conventional breeding techniques in all areas. Biochemical and Molecular techniques have shortened the duration of breeding programs from years to months, weeks, or eliminated the need for them all together. The use of molecular markers in conventional breeding techniques has also improved the accuracy of crosses and allowed breeders to produce strains with combined traits that were impossible before the advent of DNA technology
Rice is one of main food crops in the worlds so knowing about how is origin may important to the people engage in the agriculture extension or advisory or education
Yellow rust seminar by Priyanka (Phd Scholar Genetics and Plant Breeding CSK ...Priyanka Guleria
This seminar explains about the yellow rust disease of wheat: Its genetics and prevention methods as well as molecular techniques to combat yellow rust
When breeding diploid potatoes, tetraploid progeny can result from the union of 2n eggs and 2n pollen in 2x-2x crosses. Thirty-three crosses were made to examine tetraploid progeny frequency in 2x-2x crosses. All crosses were between S. tuberosum dihaploids and diploid self-compatible donors, M6 and DRH S6-10-4P17. Using chloroplast counting for ploidy determination, the frequency of tetraploid progeny was as high as 45% in one of the 33 crosses. Based upon single nucleotide polymorphism (SNP) genotyping, the tetraploid progeny were attributed to bilateral sexual polyploidization (BSP), which is caused by the union of 2n egg and 2n pollen. Dihaploids were identified that produce lower frequencies of 2n eggs. The results of this study suggest that S. tuberosum dihaploids with a high frequency of 2n eggs should be avoided in 2x - 2x crosses for diploid breeding programs.
Deployment of broad spectrum resistance against rice blast which includes gene pyramiding, deployment, transgenic approaches, marker assisted back cross breeding, pedigree by using major R genes and QTLs and phytoalexin genes.
Research Program Genetic Gains (RPGG) Review Meeting 2021: From Discovery to ...ICRISAT
Chickpea (Cicer arietinum) is the second most widely grown legume crop after soybean, accounting for a substantial proportion of human dietary nitrogen intake and playing a crucial role in food security in developing countries. We report the∼ 738-Mb draft whole genome shotgun sequence of CDC Frontier, a kabuli chickpea variety, which contains an estimated 28,269 genes. Resequencing and analysis of 90 cultivated and wild genotypes from ten countries identifies targets of both breeding-associated genetic sweeps and breeding-associated balancing selection. Candidate genes for disease resistance and agronomic traits are highlighted, including traits that distinguish the two main market classes of cultivated chickpea—desi and kabuli.
Screening of Maize Genotypes/Hybrids for Identifying Resistant Sources agains...Premier Publishers
The research included field screening of maize genotypes/hybrids for resistance to banded leaf and sheath blight (BLSB). The field screening experiment was done in a sick plot maintained at National Maize Research Program (NMRP) of Nepal Agricultural Research Council, Rampur, Chitwan. Fifteen maize genotypes/hybrids were evaluated in the field and replicated three times in randomized complete block design (RCBD). First disease incidence was observed on 60 days after sowing (DAS). Highest disease severity (81.33±1.67) was found in RL-36/RL-197. RML-76/RL-105 and RAMPUR HYBRID-2 had lowest disease severity (66.00±2.00). Also, lowest area under disease progress curve (AUDPC) value (631.67±18.33) was found in RML-76/RL-105, which was at par with P-3533 (660.00±26.67). Cob weight (r = 0.64), cob diameter (r = 0.51) and thousand grain weight (r = 0.50) were negatively correlated with total AUDPC. Total AUDPC value contributed 41.6 percent loss in cob weight, 26.2 percent loss in cob diameter and 25.5 percent loss in thousand grain weight.
Banoth Madhu: Marker Assisted Breeding in Rice. Advances in genetic studies have developed molecular techniques, thereby allowing marker-assisted breeding (MAB) for improved eating and cooking qualities in rice. MAB has gained the attention of rice breeders for the advantages it can offer that conventional breeding cannot.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
The increased availability of biomedical data, particularly in the public domain, offers the opportunity to better understand human health and to develop effective therapeutics for a wide range of unmet medical needs. However, data scientists remain stymied by the fact that data remain hard to find and to productively reuse because data and their metadata i) are wholly inaccessible, ii) are in non-standard or incompatible representations, iii) do not conform to community standards, and iv) have unclear or highly restricted terms and conditions that preclude legitimate reuse. These limitations require a rethink on data can be made machine and AI-ready - the key motivation behind the FAIR Guiding Principles. Concurrently, while recent efforts have explored the use of deep learning to fuse disparate data into predictive models for a wide range of biomedical applications, these models often fail even when the correct answer is already known, and fail to explain individual predictions in terms that data scientists can appreciate. These limitations suggest that new methods to produce practical artificial intelligence are still needed.
In this talk, I will discuss our work in (1) building an integrative knowledge infrastructure to prepare FAIR and "AI-ready" data and services along with (2) neurosymbolic AI methods to improve the quality of predictions and to generate plausible explanations. Attention is given to standards, platforms, and methods to wrangle knowledge into simple, but effective semantic and latent representations, and to make these available into standards-compliant and discoverable interfaces that can be used in model building, validation, and explanation. Our work, and those of others in the field, creates a baseline for building trustworthy and easy to deploy AI models in biomedicine.
Bio
Dr. Michel Dumontier is the Distinguished Professor of Data Science at Maastricht University, founder and executive director of the Institute of Data Science, and co-founder of the FAIR (Findable, Accessible, Interoperable and Reusable) data principles. His research explores socio-technological approaches for responsible discovery science, which includes collaborative multi-modal knowledge graphs, privacy-preserving distributed data mining, and AI methods for drug discovery and personalized medicine. His work is supported through the Dutch National Research Agenda, the Netherlands Organisation for Scientific Research, Horizon Europe, the European Open Science Cloud, the US National Institutes of Health, and a Marie-Curie Innovative Training Network. He is the editor-in-chief for the journal Data Science and is internationally recognized for his contributions in bioinformatics, biomedical informatics, and semantic technologies including ontologies and linked data.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
5. What is a doubled haploid plant?
Each cell contains 2 sets of genetic
information which are
(but not exactly) identical most.
For example, one gene set may carry a
gene for disease resistance when the
other set does not.
Doubled haploid plant has
cells containing 2 gene sets
which are exactly identical.
If one gene set has the disease
resistance gene the other gene
also having resistance.
6. History
• Blakeslee et al. (1922) - Datura stramonium
• Guha and Maheswari (1964) - Anther culture technique
for the production of haploids in the laboratory
• Niizeki and Oono (1968) - Production of rice haplpoids
• wide crossing
Kasha and Kao, (1970) - Barley
Burk et al., (1979) - Tobacco
Doubled haploid methodologies have now been applied to
over 250 species - Forster and Thomas, 2007
7. 1) Haploid production
- In vitro versus in vivo
- Maternal versus Paternal
2) Haploid identification
- Markers: Morphological or molecular
- Cytological / flowcytometry
3) Genome doubling
- Colchicine
- Other
Technologies of DH production
10. Haploid identification
Figure . Flow cytometric analysis of the ploidy level. The x-axis of the
histogram represents the intensity of DNA fluorescence in relative units; the y-
axis represents the number of nuclei counted per histogram channel. (A) A
representative peak set for diploid or doubled haploid material. (B) Peaks
corresponding to a typical haploid individual.
Payam et al., 2007
16. Stages of application of anti mitotic in DH production
Anther treatment
Microspore treatment
Haploid embryo treatment
Young haploid seedling
treatment
Young haploid Root tip
treatment
17. Fig. The interaction between the colchicine concentration in the pre-
treatment medium and the duration of colchicine pre-treatment on the
ELSs production in the ETH-M82 genotype.
Payam et al., 2007
18. Fig; Haploid embryos in cotyledonary stage (a and b)
normal regenerated plant (c) normal doubled
haploid plant (d)
Haploid embryos treated with colchicine and inoculated in
colchicine free NLN-13 medium for regeneration
Payam et al., 2011
19. Table : Effects of colchicine concentration and duration
treatment on regeneration and recovery of doubled
haploid plants of oilseed rape.
Colchicine
treatment
concentration
(mg/L)
Colchicine
treatment
duration (h)
Number of
regenerated
plants
Number
of doubled
haploid plants
0 (control) 0 53 0
125 12 49 10
24 37 9
36 18 8
250 12 41 13
24 42 27
36 15 9
500 12 17 7
24 14 8
36 9 6
1000 12 0 0
24 0 0
36 0 0
20. Table :percentage rates of doubled haploid (DH) plants
derived from individual treatments
Genotype Total no.
of plants
tested
mean Colchicine
in vivo
Colchicine
In vitro
Oryzalin
In vitro
Trifluralin
In vitro
SL-3/04 560 55.53 71.09 76.62 86.02 80.24
OP-41/1 615 39.34 55.65 77.29 69.80 88.07
SL-2/04 534 31.88 39.94 68.38 43.78 88.67
Mean 1709 42.25 55.56 74.10 66.53 85.66
Miroslav et al., 2008
22. Genetics of doubled haploid populations
• Only two types of genotypes – pair of alleles – A ,a
- Frequency – ½ AA and ½ aa
- Diploid – ¼ AA,1/2Aa,1/4aa
• Probability of getting desired genotype is (1/2)n
- Diploid – (¼)n
Kunzel et al., 2000
23. • Selfing in autogymous spp.
• Vegetative propagation
• Bud pollination
• Late or early pollination methods
26. Difference between DH method and conventional method.
Particulars DH method Conventional method
Time required for
developing pure line
1 year or 1 crop
season
3-5 year
Time required for
cultivar development
2-3 year 7-8 year
Fixation of heterosis possible Not possible
Expenditures More than
Conventional method
Less than DH method
Identification of
recessive mutation
Very easy Difficult
Singh., 2007
27. Applications of DHs in Plant Breeding
Mapping Quantitative Trait Loci (QTL)
Backcross breeding
Bulked segregant analysis (BSA)
Hybrid sorting
Genetic maps
Genetic studies
Elite crossing
Cultivar development
Fixation of heterosis
28. QTL MAPPING
Type of
Population
Strength Weakness
F2:3
- Speed of production
- d and a estimates
- Heterogeneous families
RIL - Homogeneous families
- Power of QTL detection
- Slow production
DH - Speed of producing
homogeneous families
- Power of QTL detection
- Laborious production process
- Lower recombination (>RIL)
BC - Speed of production - Heterogeneous families
29. QTLs controlling six traits determining root morphology and
distribution in a IR6429 X Azucena doubled-haploid rice population
Yadav et al., 1997
34. Phenotypic performance of parents and the DH population
for biomass yield (BY), straw yield (SY) and grain yield(GY) in
two growth seasons
Liu et al., 2006
44. Bulked segregant analysis (BSA)
• BSA is dependent on accurate phenotyping and the DH
population has particular advantage in that they are true
breeding and can be tested repeatedly.
• DH populations are commonly used in bulked segregant
analysis, which is a popular method in marker assisted
breeding. This method has been applied to rapeseed and
barley.
45. A linkage map of RAPD markers in a DH population drived
from 'Quantum‘ X 'China A' cross.
Samizadeh et al., 2007
46. PCR profiles produced by RAPD analysis in B. napus with PL
18 (A) and PL 2 (B) primers.
Samizadeh et al., 2007
47. Analysis of variance for pod length marker
Samizadeh et al., 2007
Marker MS R2 Mean ± SE
Long Short
PL2 2202.4 14.60 105.5 ± 22.01 92.10 ±10.03
PL9 712.42 4.70 101.6 ± 20.33 94.10 ±13.27
PL 14 863.62 6.00 102.2 ± 20.22 93.90 ±13.56
PL 17 522.39 3.40 101.7 ± 18.98 95.14 ± 16.12
PL 18 1468.3 9.70 105.2 ± 19.46 93.94 ±12.24
PL 26 1027.4 7.80 100.6 ± 18.98 89.94 ± 8.19
PL 2 × PL 18 1129.6 22.40 112.2 ± 21.86 89.23 ± 5.23
48. Hybrid sorting
• Hybrid sorting - Selection of superior plants among hapliods
derived from F1 through anther culture
• Selection of recombinant superior gametse
• Superior over pedigree & bulk method
- frequency of superior gametes – higher than the
corresponding F2 generations
- Reduces the time required to release a variety
• Successful in china & Japan
49. Varieties bred through hybrid sorting
crop Varieties developed Attributes
Rice Tanfong 1, xin xion,
late keng 76,
shanyou 63
Good quality, high
yield
Wheat Yunhua 1, yunhua 2 Rust resistance,cold
resistance,lodging
resistant
Tobacco Tanyu 1, Tanyu 2,
Tanyu 3
Disease resistant,
Mild smoking
50. SELECTION OF SALT TOLERANCE GENOTYPES FROM
DOUBLED HAPLOIDS IN RICE
Dang et al., 2004
51. Table : Relative response of anther culture lines and
the tolerance checks at salinity of 6dS/m
and 15dS/m.
52. Rice anther culture to obtain DHs with multiple resistance
Bambang et al., 2010
• Released varieties: Way Rarem, Jatiluhar.
• Accessions tolerant to aluminum toxicity( Al): Dupa, Krowal
• Accessions tolerant to shade: Dodakan, ITA-247
Results
• 5 lines – tolerant to Al toxicity, and shade and tolerant to
4 races of blast.
• 11 lines – tolerant to Al toxicity, and tolerant to 4 races
of blast.
• 1 lines – tolerant to shade and tolerant to 4 races of
blast.
• 2 lines – sensitive to Al toxicity, and shade but tolerant
to 4 races of blast.
53. Linkage maps of F2 populations derived from the
cross between two temperate japonica cultivars
‘Koshihikari’ and ‘Akihikari’
Masumi Yamagishi et al.,2010
54. Linkage maps of DH populations derived from the cross between
two temperate japonica cultivars ‘Koshihikari’ and ‘Akihikari’
Masumi Yamagishi, et al
55. Genetic studies - mutation genetics
• Genetic ratios and mutation rates can be read directly
from haploid populations.
• A small doubled haploid (DH) population was used to
demonstrate that a dwarfing gene in barley is located
chromosome 5.
• In another study the segregation of a range of markers
has been analyzed in barley.
56. Means ± standard errors and ranges for erucic acid (% of total fatty
acids) in seeds of the M2 and M3 generations of six doubled haploid
mutant lines of Brassica carinata Barro et al.,2002
Line Plants analysed Generation
M1 M2
Control 10 42.8 ± 0.7 43.5 ± 0.5
40.2–44.4 41.4–44.7
BC2.6.1 8 48.5 ± 0.3 50.6 ± 0.4
47.8–48.7 49.7–51.5
BC2.8.1 6 48.3 ± 0.4 49.6 ± 0.2
48.1–48.9 49.4–49.9
BC3.3.2 8 48.1–48.9 50.3 ± 0.6
47.8–49.8 49.2–51.1
BC5.2.2 10 48.7 ± 0.4 49.7 ± 0.3
48.3–50.1 49.1–50.7
BC6.19 8 49.5 ± 0.6 48.5 ± 0.2
48.8–50.2 48.0–48.8
BC1.5 10 48.7 ± 0.6 48.7 ± 0.1
48.4–49.6 48.5–48.9
57. Dendrogram showing the relationship among 102 DH
wheat based on gliadins bands.
Ojaghi and Akhundova., 2010
59. Figure . Dendrogram showing the relationship among 102 doubled
haploid wheat based on RAPD
markers
60. Doubled haploid varieties
Crop Varieties Country
Rice Xin-Xin, Hua-Hau-Zao china
Wheat Jing Hua 1,3,5 china
Barley Mingo Canada
Chickpea Quantum, Q2, Duplo, Mingo
Cabbage Orange queen Japan
Brocolli Three man Japan
Tobacco Dan-yu1,2,3 China
Hot pepper Haihua 19 China
Sweet pepper Haihua 29 China
Raina ,1997
61. Elite crossing:
• Traditional breeding methods are slow and take
10–15 years for cultivar development.
• Another disadvantage is inefficiency of selection in
early generations because of heterozygosity.
• These two disadvantages can be over come by DHs,
and more elite crosses can be evaluated and
selected within less time.
• Improved genetic gain can be achieved with DHs .
62. Obtained DH lines from spring and winter
wheat hybrids
Grauda et al.,2010
63. Application of Doubled haploid in Cultivar
development
Singh., 2007
Name of crop No of cultivar released Developed by
Barley 115 Anther culture
Rape seed 47 Anther culture
Wheat 21 Anther culture
Melon 9 irradiation
Capsicum annum 8 Anther culture
Rice 8 Anther culture
Asparagus 7 irradiation
Tobacco 6 Anther culture
Egg plant 5 Anther culture
65. Double haploid rice varieties in India.
AICRIP and PH-43
has performed well (2000- 05)
CRAC 2224 -1041 (early duration )
PHB71, PA6201, DRRH1, KRH2, Pusa RH 10,
CRHR 4 and Rajlaxmi
2000 DH lines developed
69. Speed in Line Development
Founder line 1 x Founder line 2
F1
Selfing
Inbred line
DH
Inbred line
70. Advantages
of Doubled Haploid Techniques
• No risk of herterozygosity - Based on gamete selection
• Develop immediate homozygosity, shorten the time to cultivar release -
additive & additive x additive variances
• Provide greater efficiency of selection in plant breeding
cytogenetics
• Production of aneuploids & determine the basic chromosome number
• Improve the precision of genetic and mapping studies
• Accelerate gene pyramiding
• Improve efficacy and efficiency in screening for resistance
71. Some Drawbacks
with Doubled Haploid Production
GENERAL:
– More expensive: expertise, facilities
– Restriction on number of crosses
• SPECIFIC:
– Mutagenic treatment
– Genotype dependent haploid induction
– Low haploid regeneration frequency