1. Assignment
On
Advances in Plant Breeding System
GPB 903
Submitted To
Prof. (Dr.) Shailesh Marker
Department of GPB
Submitted by
Divya Singh
ID. No: 18PHCBC101
Department of BCBE
JIBB
Department of Biochemistry & Biochemical
Engineering, JIBB,
Sam Higginbottom University of Agriculture, Technology
& Sciences Prayagraj,
U.P., India
2019
2. (AI-Daej et al.,2019)
Biotechnology and related tools to support conventional breeding strategies, to
overcoming the limitations in crop production and improving quantity and
quality as well as climatic and disease stress tolerance of the crop advance
plant breeding is going on in this time. There is need to improve the
conventional breeding to advance plant breeding.
By various genomics Modern tools :
Molecular markers
Initially low density for QTL mapping, introgression of major genes into elite
germplasm with high-density markers, association mapping and
MAS/genomic selection
New statistical tools
–Mixed model methods
– Bayesian approaches to handle high-dimensional data sets
– New methods to deal with G x E
3. Other technologies
Better standardization of field sites (laser-tilled fields, GPS, better micro-
and macro-environmental measurements)
• High throughput phenotypic scoring
• DH lines
• GC/MS
• LC/MS
• HPLC
Biotechnological approaches such as RNA interference (RNAi),
microRNA, Clustered Regularly Interspaced Short Palindromic Repeats
(CRISPR)/CRISPR-associated system (CAS), transcription activatore like
effector nucleases (TALEN), zinc-finger nuclease are being used to
introduce the nutritionally important potential genes through genetic
engineering (introducing or editing the genome).
(Fentik et al., 2017)
11. Among the diverse wild gene pool of okra, it holds resistant source of gene for
many biotic stresses. However, solving the problem of diseases and pests
infestation in okra by traditional breeding approach is one of the value added
steps for resistant variety released in the recent past. Now, the advancement in
molecular and biotechnological techniques enhances the okra improvement
programmes by advancing in marker assisted selection and resistant gene
transfer.
Vitamins,
Minerals,
Calcium,
Carbohydrates,
Proteins
Dietary fibre
Edible oil (20 to 40%)
High constitute of linoleic acid (48%) which have health benefits for
patient suffering from cardiovascular problems.
(Chowdhury et al., 2019)
12. Recent advancement in the molecular biology leads more reliable
identification of cultivars with suitable traits, genetic diversity and
prediction of heterosis among germplasms.
DNA markers are extensively utilized now-adays by the researchers to
acquire quick, cost effective, reliable and efficient outcome irrespective of
environmental alteration.
Introduction of various molecular markers such as RAPD, SSR, EST-SSR
and ISSR is more effective way to exploit heterosis process when
congruent with phenological characters.
The transcriptome profiling of YVMV susceptible verites are- A. manihot
(HR), A. angulous (HR) , A. pungens (HR) , A. moschatus (MR)
13. These genes are governed to influence defense mechanisms against
diseases in okra species.
Okra enation leaf curl virus (OELCV) in cultivated lines of okra are
A. tetraphyllus (R), A. ficulneus (HR) A. crinitus (HR), A. angulous
(HR), A. pungens (HR), A. tuberculatus (R), A. moschatus (MR).
Advancement in the molecular technology
Marker assisted selection,
Genome wid sequencing,
Next generation sequencing
15. Evaluation of advance
lines:
Forty advance lines were
evaluated during kharif season
for yield, dark green fruit,
devoid of seed bulging and
reaction to viral disease like
YVMV and ELCV. Genotype
VRO-120 (Fruit yield/plant:
300g;) and VRO-125 (Fruit
yield/plant: 360g;) identi ed
as most promising genotypes
for fruit yield, fruit quality
and disease resistance during
eld evaluation of advance
lines.
(ICAR-Indian Institute of Vegetable Research 2019)
16. Molecular characterization of Okra Enation Leaf Curl Virus
RNA interference-based resistance in transgenic okra plants against
Okra Enation Leaf Curl Virus (OELCV) and its associated
betasatellite: The clone of Okra enation leaf curl beta was used as
template for PCR ampli cation of βC1 gene for further analysis.
Genome editing in Okra: CRISPR/Cas9 mediated genome editing work
in okra was undertaken to progress the work towards the development of
coat protein construct for plant transformation. The guide RNA construct
targeting replicase (rep) and coat protein (cp) gene of Okra Enation Leaf
Curl Virus (OELCV) was arti cially synthesized. The target region of
replicase and coat protein gene was identi ed manually from a conserved
sequence by multiple sequence alignment of major strains of OELCV to
achieve broad spectru resistance. Complementary gRNA oligonucleotide
pairs will be annealed and cloned into pORE-04 vector by Ligation.
In-plant transformation in okra: In dependent, in planta transformation
of okra in the cultivar Kashi Kranti was repeated. For Agrobacterium
mediated in plant transformation.
(ICAR-Indian Institute of Vegetable Research 2019)
17. Rice (Oryza sativa L., 2n = 2x = 24) is second most widely grown cereal
crop and the staple food for more than half of world’s population,
providing two thirds of calorie intake for more than three billion people in
Asia and one-third of calorie intake of nearly 1.5 billion people in Africa
and Latin America.
It is probably a descendent of wild grass that was most likely cultivated in
the foothills of the far Eastern Himalayas.
Some believes that the rice plant may have originated in southern India,
then spread to the north of the country and then onwards to China.
(Khan et al., 2015)
19. Blast disease is one of the major pathogens affecting crop productivity.
Breeding for resistance is economic way to produce verirty and tissue
culture used as a tool to decrease the period of breeding program.
Seven varieties(AI-Asha Type 1, AI-Asha Type 2) Sakha 104, Gz6903,
Giza 177 and Sakha 105) were used for tissue culture and crossing
through line X tester method.
Somaclonal variation and 12 F1 were evaluated and estimated the GCA
nad SCA under field condition. The MS with 2,4-D (2,4
Dichlorophenoxyacetic acid) 3 % sucrose were used for callus induction
and plant regeneration.
21. For callus induction, the varieties Sakha 104,Gz6903, Giza 177 and
Sakha 105 gave 100%, while, varieties AI-Asha Type 1, AI-Asha Type 2
gave 85%. In addition, 18 lines produced from AI-Asha Type 1 result
showed that 13 line were resistant to blast, while 6 line were susceptible.
As for SC1 family derived from Sakha 104, 11 Line were produced and
results showed that 6 lines were susceptible to blast and five line were
resistant to leaf blast.
22. Tissue culture in traditional plant breeding is one of the most effective
means for production new varieties of rice resistant to blast disease as well
as improving crop traits.
The most promising hybrid combinations were
I. Type 1 X Sakha 105,
II. AI- Ahsa Type2 X Giza 178,
III. AI-Ahsa Type2 X Sakha 105,
IV. Sakha 104 X Giza 177
V. Gz6903-1-2-2 X Sakha 105
for desirable traits and could be utilized in rice breeding program to improve
these traits.
23. (Kumar et al., 2019)
Embryo culture
Callus induction and Regeneration
Somatic Hybridization
Effect of EMS
EMS is considered as one of the most important chemical mutagenic
Agents and it is considered as an alkylating agent that has a mutagenic effect
on the DNA.
Genetic variability apart from conventional breeding can be easily discernible
in in-vitro regenerated somaclonal variants.
Tissue culture techniques through mutation induction can be used to increase
the speed or efficiency of breeding programs to get a new diversity of
germplasm. In vitro mutation induction can be done by using chemical
mutagens and also with use of growth regulators which have high activity.
24. (Kumar et al., 2019)
Phenotypic Characterization of EMS Induced Mutants in Wheat (Triticum aestivum L.)
Effect of UV light
Reported that UV-B light in agriculture systems constitutes a promising tool to
increase crop production and protection against pests. contributed to deeper
understanding of influence of UV - rays on plant growth, development and
metabolism.
Reduction in growth of wheat primary leaves was found to be due to an effect
of UV-B on the rate and duration of both cell division and elongation. In UV-
B-grown plants, the proportion of mitotically active cells was reduced, and the
duration of cell division increased.
Thus, the supply of cells into the elongation zone was reduced in UV-B-grown
plants, which, coupled with a reduction in the rate of elongation, resulted in the
observed reduction in leaf growth.
Directorate of Wheat Research Karnal (Haryana)
25. Traits Studied
Phenology
Days to heading
Days to Maturity
Colour/pigmentation
Height
Floral mutants
Spike length
Symmetry
Threshability
Floral parts
Yield Traits
Color
Size
Shape
Directorate of Wheat Research Karnal (Haryana)
26. Open floret, deformed glume, twisted awns etc..
Spikelet symmetry variations
ABCDE model of flower development
Directorate of Wheat Research Karnal (Haryana)
27. Threshability: Easy and Hard
Free threshing Q gene mutants in wheat Q gene copy no variations
Range: 2 -18 cm
Directorate of Wheat Research Karnal (Haryana)
28. Shrivelled and unfilled grains
Beak Shaped Grain mutant
Directorate of Wheat Research Karnal (Haryana)
29. EFFECT OF EMS ON ANTIOXIDANT ENZYME ACTIVITY AND
OXIDATIVE STRESS OF TRITICUM AESTIVUM L. VAR HD-2894
Ethyl methano sulphonate (Ems), a chemical mutagen is widely used to induce a
large number of functional variation in wheat plant. The objective of present study
was to determine the activity of enzymatic antioxidants and Lipid peroxidation
(LPO) after EMS exposure in a wheat plant in M1, M2 and M3 generation of
Triticum aestivum L. var. HD 2894
Presoaked seeds were treated with EMS for 6 hour with different concentration like
0%, 0.1%, 0.2%, 0.3%, 0.4% and 0.5%. Some biochemical parameter such as
Catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR) and Lipid
peroxidation (LPO) were studied.
One the conclusion by the EMS treatment in each generation in M1 – CAT
decrease M2- CAT increased M3- CAT decreased the M2 generation, APX
decreased in M1, in M2- APX in Increased by giving different treatments, M3
generation it show stability , GR activity in M1 generation is decreased but in M2 it
is increased and in M3 increased, LPO in M1 generation it decreased in M2
increased and in M3 also increased it seen in deferent absorbance at different
concentration.
Dubey S. J.A. et al., (2017)
42. Identified altered phenotypes for a large number of traits
Role in breeding for wheat improvement (High Tiller and Bold grain
mutants)
Understanding the functional basis of genes
Openness of spikelet’s and male sterility could possibly find a role in
hybrid wheat development
Dubey S. J.A. et al., (2017)
43. Genomics can complement the breeding for quality improvement in
sorghum and millet. Good progress has been made in improving grain
quality traits in sorghum through use of biotechnology for biofortification
which can support crop improvement for better nutrition. Integrating
marker-assisted breeding (MAB) with classical breeding enables the
possibility to track the introgression of nutritional quality-associated QTLs
and genes into popular cultivars from various germplasm sources.
Nutritional quality traitsdprotein content, amino acid composition, fat and
starch contents, and density of micronutrients. Identification of genes
influencing sorghum grain composition would help manipulate grain
texture and quality to accommodate existing end-use markets and promote
new product development.
(Parthasarathy Rao et al., 2006; Ashok Kumar et al.,
2011, 2013)
45. Key discoveries in the genetic control of grain composition through sorghum
mutations include an amylose QTL which is 12 kb away from the waxy locus,
the primary gene regulating amylose production and improved starch
digestibility, sugary which has increased sucrose content, and high-lysine,
which increases lysine content and protein digestibility .
Recent phenotyping developments have resulted in high-throughput
phenotyping platforms that utilize proximal sensing to measure multiple
physiological traits.
Determining food mineral composition is essential in order to understand its
nutritional value showed that atomic absorption spectroscopy was effective in
detecting several metals at parts per million level of concentration. However,
mineral profiling techniques at tissue level are time-consuming, hence the
growing demand for development of modern analytical techniques to help
increase plant yields and quality.
(Rhodes et al., 2017; Kotla et al., 2016)
46. Nuclear magnetic resonance spectroscopy and mass spectrometry (MS) coupled
with gas chromatography (GS/ MS) or liquid chromatography (LC/MS) are being
used to identify plant metabolites such as amino acids, lipids, and carbohydrates
(primary metabolites) and polyphenols, alkaloids, terpenes, polyketides, and
hormones (secondary metabolites) in sorghum.
In sorghum, high-performance liquid chromatography (HPLC) is frequently
utilized in phytonutrient phenotyping of phenolic compounds and flavonoids.
marker-trait associations (MTAs) have been found, whereby six genes involved in
synthesis pathways of starch (Sh2, Bt2, SssI, Ae1, and Wx) or grain storage
proteins (o2) showed strong association with different grain quality traits, two yield
component traits, and yield.
(Bean et al., 2016)
47. Fig- Positions and Descriptions of QTLs Affecting Grain Fe and Zn
Concentration in the (ICMB 841-P3 863B-P2)-Derived RIL Population
Across the Two Environments at ICRISAT
(Bean et al., 2016)
48. Additionally, both gas and liquid chromatography can be simultaneously
applied, as has, for example, been performed in rice seeds where their
antioxidant properties were compared with the content of metabolites .
Furthermore, the technique of ultraperformance liquid chromatography
provides better peak separation and higher reproducibility of retention time
when compared with HPLC and, when coupled with quad timeof- flight
(Q-TOF), its sensitivity increase.
(Fatima et al., 2015, Guo et al., 2011)
50. Recent advances in sequencing techniques provide an opportunity for the
precise detection of mutagenesis-induced sequence variations at a large
scale in the genome.
Next-generation sequencing based mutation mapping approaches including
Mutmap, MutChromeSeq, and whole-genome sequencing-based mapping
which has enormous potential to accelerate the mutation breeding in
tomato.
The demand for tomatoes is increasing day by day but as its production is
affected by many diseases and stresses (biotic and abiotic stress).
Genetic engineering techniques can also play an important role in the
improvement and development of disease resistant cultivars. So, now days
it is becoming obvious that improvement of this crop is a critical task to
overcome the limitation of tomato production. (Fentik et al., 2017)
51. Mutation breeding - Mutation breeding now days is used as an important
tool to develop a variety in a very short time by the breeders. When the
mutation is induced by using various chemicals for crop improvement is
called mutation breeding.
About 6000 Micro-Tom mutant lines were generated using gamma-rays
irradiation. From this, around 24 morphological mutant lines and eight brix
mutant lines were screened and QTL analysis revealed the involvement of
two loci in the brix mutants which can be used to identify brix-regulating
genes in tomato. Similarly, fast neutron and other physical mutagens can be
explored to induce beneficial variability in tomato for enhanced quality and
yield.
Zinc finger nucleases (ZFNs)-based genome editing is one of the primitive
technologies which makes it possible to perform precise site-specific
mutations. The ZFNs form dimmers which recognize a specific target site
and make a double-strand DNA break.
Chaudhary J. et al.,(2019)
57. Schematic representation of
atypical mutmap strategy
exploring next generation
sequencing to identify the
causal mutations resulted
into altered phenotype in
plants.
Chaudhary J. et al.,(2019)
59. Generalised flowchart showing steps involved in the MutChromSeq
approach which can be conveniently used for the mapping of casual
mutation in plant species with larger genome size.
To selecchromosome for the sequencing, first mapping of confirmed
mutant need to be done with markers known to be linked with each
chromosome.
Subsequently, chromosomes labelled with fluorescence dye can be sorted
with flowcytometric techniques and sequenced selectively with high
throughput techniques.
Later analysis of sequencing data from the mutant line can be used to
locate the casual mutation precisely similarly as described in Figure 1
Chaudhary J. et al.,(2019)
60. Conclusion
It is now rightly said that the population of the country is increasing day by
day and to meet the requirement of the whole country breeder need to
develop the high yielding varieties which is not possible with the
traditional breeding methods. Use of molecular marker techniques are
establishing biotechnological approaches (RNAi), microRNA,
(CRISPR)/CRISPR- (CAS), (TALEN), zinc-finger nuclease are being used
to introduce the nutritionally important potential genes through genetic
engineering (introducing or editing the genome) therefore, the combined
application of traditional breeding and plant biotechnology methods
including selection based on molecular markers marker-Assisted Selection
might be valuable tools for Advanced plant breeding.
(Fentik et al., 2017)
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in Tomato: Advances, Applicability and Challenges Department of Biology, K.) National
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