The document summarizes a seminar presentation on breeding for different flower forms in ornamental crops. It discusses the ABCDE flower development model and how genes in this model control formation of floral organs. It also covers conventional breeding techniques like hybridization and inheritance patterns for double flower traits. Mutation breeding methods like physical and chemical mutagens are explained, along with examples of mutants induced in different ornamental crops. Polyploidy induction using colchicine is also summarized, with a case study on inducing polyploidy in Gladiolus grandiflorus. The presentation covers various genetic modification techniques for creating novel flower shapes in ornamental plants.
Selfing and crossing techniques in crop plants and Breeders KitRajendragouda Patil
Studying about, Selfing and crossing techniques in crop plants and Breeders Kit,helps to students and researchers to select appropriate techniques for selfing and crossing in different crops.
Plant breeding methods of vegetatively propagated crops Roksana Aftab Ruhi
Vegetatively propagated crops are bred by intentionally crossing of closely or distantly related individual to produce new crop varieties or lines with desirable traits. Breeding of vegetative crops have successfully improved quality, yield, tolerance of crops to environmental pressure. Breeding helps in producing crops that are resistant to viruses, fungi and bacteria and helps in longer storage period for the harvested crop.
Tuberose - introduction and uses – varieties - soil and climate and planting systems - weed, nutrition and irrigation management – special horticultural practices - role of growth regulators- harvest index and yield
Pract no. 9 (a) floral biology of bananatusharamodugu
Botanical name : Musa paradisiaca L. (Fruit variety)
Musa sapientum (Vegetable variety)
Common Name : Banana, Kela
Chromosome number : 2n = 3x = 33
Banana is one of the oldest fruits and second largest growing fruit crop in the world. It is also known as “Adams Fig “and “Apple of Paradise.”
Importance :
It is widely used as a fresh fruit.
The central core of the pseudostem is used as a vegetable.
The banana pseudostem is also used for manufacturing paper and boards.
Differential Effects of Sucrose and Plant Growth Regulator on Shoot Multiplic...drboon
Explants from young leaves and stem nodes of Oxalis versicolour were used and cultured on MS medium supplemented with different concentration of 2,4-D. The best result showed that cluster of callus were formed and proliferated around the base of explants on MS medium supplemented with 0.1 mg/l 2,4-D. Callus transferred to MS medium supplemented with various concentrations of NAA and BA. After nine weeks, callus regenerated to be new shoots. The highest average length of stolon was from MS medium supplemented with 0.1mg/l NAA and 0.1 mg/l BA and number of plantlets was from MS medium supplemented with 4.0 mg/l NAA and 5.0 mg/l BA. Plantlets were cultured on MS medium supplemented with different concentrations of sucrose for ten weeks. It was found that all parameters: number of plantlets, bulbil sized, length of stolon, and number of nodes were significant difference (p≤0.05). Number of flowers and sized of flowers found only in MS medium supplemented with 9–10 % of sucrose.
Selfing and crossing techniques in crop plants and Breeders KitRajendragouda Patil
Studying about, Selfing and crossing techniques in crop plants and Breeders Kit,helps to students and researchers to select appropriate techniques for selfing and crossing in different crops.
Plant breeding methods of vegetatively propagated crops Roksana Aftab Ruhi
Vegetatively propagated crops are bred by intentionally crossing of closely or distantly related individual to produce new crop varieties or lines with desirable traits. Breeding of vegetative crops have successfully improved quality, yield, tolerance of crops to environmental pressure. Breeding helps in producing crops that are resistant to viruses, fungi and bacteria and helps in longer storage period for the harvested crop.
Tuberose - introduction and uses – varieties - soil and climate and planting systems - weed, nutrition and irrigation management – special horticultural practices - role of growth regulators- harvest index and yield
Pract no. 9 (a) floral biology of bananatusharamodugu
Botanical name : Musa paradisiaca L. (Fruit variety)
Musa sapientum (Vegetable variety)
Common Name : Banana, Kela
Chromosome number : 2n = 3x = 33
Banana is one of the oldest fruits and second largest growing fruit crop in the world. It is also known as “Adams Fig “and “Apple of Paradise.”
Importance :
It is widely used as a fresh fruit.
The central core of the pseudostem is used as a vegetable.
The banana pseudostem is also used for manufacturing paper and boards.
Differential Effects of Sucrose and Plant Growth Regulator on Shoot Multiplic...drboon
Explants from young leaves and stem nodes of Oxalis versicolour were used and cultured on MS medium supplemented with different concentration of 2,4-D. The best result showed that cluster of callus were formed and proliferated around the base of explants on MS medium supplemented with 0.1 mg/l 2,4-D. Callus transferred to MS medium supplemented with various concentrations of NAA and BA. After nine weeks, callus regenerated to be new shoots. The highest average length of stolon was from MS medium supplemented with 0.1mg/l NAA and 0.1 mg/l BA and number of plantlets was from MS medium supplemented with 4.0 mg/l NAA and 5.0 mg/l BA. Plantlets were cultured on MS medium supplemented with different concentrations of sucrose for ten weeks. It was found that all parameters: number of plantlets, bulbil sized, length of stolon, and number of nodes were significant difference (p≤0.05). Number of flowers and sized of flowers found only in MS medium supplemented with 9–10 % of sucrose.
The production of haploid plants exploiting the totipotency of microspore.
Androgenesis is the in vitro development of haploid plants originating from totipotent pollen grains through a series of cell division and differentiation.
The term genetic pollution was popularized by environmentalist Jeremy Rifkin in his book “The Biotech
Century”(1998).Genetic pollution accounts to the uncontrolled spread of genetic information (frequently
referring to transgenes) into the genomes of organisms in which such genes are not present in nature.
Genetically engineered (GE) plants contains genes which have been transferred from unrelated species. These
may come from bacteria, viruses,other plants or even animals. If these ‘foreign’ gene are the n transferred into
other organisms,this causes genetic contamination or pollution of the natural genepool.
Genetic pollution is usually associated with the gene flow from a genetically engineered
(GE)organism(orgeneticallymodifiedorganism-GMO)toanon-GMorganism."Geneticpollution"andcollateral
damage from GE field crops already have begun to wreak environmental havoc. Wind, rain, birds,bees, and
insect pollinators have begun carrying genetically-altered pollen into adjoining fields, polluting the
DNAofcrops oforganicand non-GEfarmers’
DEFINITION
The dispersal of contaminated or altered genes from genetically engineered organism to natural
organism.
"Uncontrolled spread of genetic information (frequently referring to transgenes) into the genomes of
organisms in which such genes are not present in nature”
Gene flow
Gene flow is the movement of genes from one population to another, conferring new traits – the biophysical
characteristics of the organism – to individuals of the recipient population. This happens by cross-pollination
(also called hybridisation), that is, the pollination of members of one population or genetic pool with that of
another.
Transgene flow
Transgene flow is the loss of potentially useful crop genetic diversity in the recipient population (whether
other crops, landraces or wild relatives). Outbreeding depression (the reduction of fitness from hybridisation)
can lead to a decrease in allelic diversity by extinction of members of a diverse gene pool that are less adapted
to survive because of the particular introgressedtransgenic trait .
The ethanol extracts of Ficus asperifolia, Mormordica charantia, Anacardium
occidentals and Psidium guajava were evaluated sole and in treatment combinations at 25, 50 and
75mg ml-1 concentration levels against the mycelial growth of Macrophomina phaseolina of
Cowpea. The pathogen was cultured on plates containing botanicals amended Potato Dextrose
Agar (PDA) in three replicates while only ethanol treated PDA tested plates served the control
experiment. The radial growths were recorded at 4th, 6th and 8th day after inoculation. Data
obtained were analysed using the SAS software program version 9.2. The extract of Mormordica
charantia was the most effective in the botanical treatments alone. The most significant inhibition
of Macrophomina phaseolina were observed from the combined treatments of Ficus asperifolia,
Mormordica charantia and Anacardium occidentals (3.11 cm), followed by Mormordica
charantia and Psidium guajava (3.29 cm), then combination of four extracts; Ficus asperifolia,
Mormordica charantia, Anacardium occidentals and Psidium guajava (3.53 cm), then
Mormordica charantia and Anacardium occidentals (3.84 cm). Other treatments, either alone or in
combination produced significant result compared to the control experiment (6.94 cm). However,
the efficacy of botanicals increased with concentration and also significantly correlated with time
and reduction in mycelia extension of the pathogen. More so, variability in the antifungicidal
potentials of the botanicals on Macrophomina phaseolina ranges from 15.93% to 34.06%
according to Eigen proportions. The treatment combinations of; Ficus asperifolia, Mormordica
charantia and Anacardium occidentals at 75mg ml-1 concentration level produced the most
inhibitory effect against Macrophomina phaseolina in vitro. However, the untreated plates did not
show inhibitory effect on the mycelial growth of the pathogen. Therefore, combined treatments of
botanicals could be a potential source in the practice of plant disease control.
Abstract
Potato is an important food and cash crop in Eastern Ethiopia; however, its productivity is low for a number of constraints. Shortage of quality planting material and poor tuber sprouting due to long dormancy period of improved varieties at planting are two of the factors known to affect production cycle and productivity of the crop in Eastern Ethiopia. Two separate experiments were conducted from November 2013 to June 2014, to assess the effect of Gibberellic acid and storage condition on seed tuber dormancy breakage of two potato varieties. The treatments in the first experiment consisted of two potato varieties (‘Bubu’ and ‘Bate’) and three levels of Gibberellic acid (GA3) (0, 10, and 20 ppm) kept under three storage methods: in diffused light store (DLS), in pit, and in farmyard manure (FYM) heap. The experiment was laid out as a randomised complete design with four replications and conducted in the horticulture laboratory of Haramaya University. The second experiment consisted of the same treatments laid out in the field to study the effects of the treatments on the subsequent growth, yield, and yield-related traits. The experiment was laid out in a randomised complete block design with three replications and conducted on a farmer’s field. The results of the experiments showed that genotypes, exogenous application of GA3, and storage conditions, as well as the interaction between them, significantly affected seed tuber dormancy period, sprouting characteristics, and subsequent tuber yield. Dormancy period, sprouting percent, sprout length, length of lateral axillary sprouts, and sprout vigour were significantly affected by the treatments. However, parameters such as days to 50% emergence, days to 50% flowering, and number and weight of very small and small tubers showed highest values for seed tubers, either treated with GA3 or not, and stored under FYM heap and pit storage conditions when compared with tuber treated and stored in DLS. In general, the study indicated that the interaction between genotypes, exogenous application of GA3, and storage conditions resulted in early dormancy termination, early emergence of shoots, and high marketable tuber yield.
Gemeda Mustefa
Mutation Induction for Improvement of Banana (Musa Spp). "Berangan Cv. Intan-...paperpublications3
Abstract: In vitro mutation induction by using gamma ray at 20, 30, 40 and 60Gy was used to generate variability in triploid banana "Berangan cv. Intan (AAA), so as to provide the opportunity to select plants with desirable characters such as early fruiting and short stature. Mutation frequency increased with increased dosage whereas, survival and capacity to regenerate decreased with increased doses. Time to initiation varied from 4-8 weeks for gamma-irradiated materials compared to 2-3 weeks in the control. It appeared that the higher the dose, the longer it took for shoot initiation. The exposure of shoot-tip meristem pieces to radiation doses produced wide variation in growth and morphogenetic performance. Mutagenic treatments induced 2 to 3- fold increases in variability in both quantitative and qualitative traits at different stages, in vitro; at nursery and field. For the field-grown plants; the proportion varied from 2.9% for the control plants to 16.8% for 60 Gy and 20.1% for 40 Gy-treatments, while for treatments at 20 and 30 Gy variations was at 18% and 19.2%, respectively. The frequency of variants was highest in 40 Gy followed by 30 Gy and 20 Gy, while it was very low in 60 Gy except for plant stature (dwarfism or stunted growth). Earliness to flowering variants were recorded at 20, 30 and 40 Gy at low frequencies (0.6%, 0.7% and 1.7%) respectively, while none was observed for 60 Gy treatments. As in vitro mutation induction could create genetic variability as well as many undesirable variants, it is highly desirable to integrate in vitro mutation with a selection system that can screen for large mutagen treated population. The useful variants recorded for earliness to flowering were selected for 30 and 40 Gy treatments. 40 Gy showed high frequency in earliness as compared with 20 and 30 Gy., which came to flowering as early as 6 – 6.5 months compared to 7-8 months for control. The useful dwarf, which considered as desirable traits, showed a balance of height and girth.
Mutation Induction for Improvement of Banana (Musa Spp.) Berangan Cv. Intan-AAApaperpublications3
Abstract: In vitro mutation induction by using gamma ray at 20, 30, 40 and 60Gy was used to generate variability in triploid banana "Berangan cv. Intan (AAA), so as to provide the opportunity to select plants with desirable characters such as early fruiting and short stature. Mutation frequency increased with increased dosage whereas, survival and capacity to regenerate decreased with increased doses. Time to initiation varied from 4-8 weeks for gamma-irradiated materials compared to 2-3 weeks in the control. It appeared that the higher the dose, the longer it took for shoot initiation. The exposure of shoot-tip meristem pieces to radiation doses produced wide variation in growth and morphogenetic performance. Mutagenic treatments induced 2 to 3- fold increases in variability in both quantitative and qualitative traits at different stages, in vitro; at nursery and field. For the field-grown plants; the proportion varied from 2.9% for the control plants to 16.8% for 60 Gy and 20.1% for 40 Gy-treatments, while for treatments at 20 and 30 Gy variations was at 18% and 19.2%, respectively. The frequency of variants was highest in 40 Gy followed by 30 Gy and 20 Gy, while it was very low in 60 Gy except for plant stature (dwarfism or stunted growth). Earliness to flowering variants were recorded at 20, 30 and 40 Gy at low frequencies (0.6%, 0.7% and 1.7%) respectively, while none was observed for 60 Gy treatments. As in vitro mutation induction could create genetic variability as well as many undesirable variants, it is highly desirable to integrate in vitro mutation with a selection system that can screen for large mutagen treated population. The useful variants recorded for earliness to flowering were selected for 30 and 40 Gy treatments. 40 Gy showed high frequency in earliness as compared with 20 and 30 Gy., which came to flowering as early as 6 – 6.5 months compared to 7-8 months for control. The useful dwarf, which considered as desirable traits, showed a balance of height and girth.
A field experiment was conducted on at M.lekhe district (Ethiopia) during 2002 and 2003 years to investigate the response of tomato to rates of Nitrogen (N) and Phosphorus (P) fertilizers. The treatment consisted of factorial combination of four Nitrogen fertilizers rates (50 kg, 100 and 150 urea/ha) and four P rates (100,150 and 200 DAP/ha) arranged in a Randomized Complete Block Design. Statistically significant and highest yield per plant was recorded at the highest rate of DAP (200 kg/ha). The significantly lowest yield was found at the zero level (with out DAP applied). The marketable yield in Q/ha of the rates is 939.96, 822.44, 731.1067 and 421.44 for 200, 150, 100 and 0 rates respectively. As the partiual budget analisis showed increasing rate of phosphorus and urea fertilizers increased profitability until 200 kg/ha and 150 kg/ha respectively.
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 .
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
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.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
3. UNIVERSITY OF HORTICULTURAL SCIENCES, BAGALKOT
2nd SEMINAR
Breeding for different flower forms in ornamental crops
Takhellambam Henny Chanu
UHS18PGD253
Department of FLA
4/22/2021 Department of FLA 3
4. OUTLINE OF PRESENTATION
➢ Introduction
➢ ABCDE Flower Development Model
➢ Conventional breeding
➢ Genetic modification
➢ Case studies
➢ Future thrust
➢ Conclusion
4/22/2021 Department of FLA 4
5. INTRODUCTION
▪ Flower shape - important
characteristics of ornamental
plants.
▪ Creation of new flower shapes -
major breeding target of any
breeder.
▪ Modification in flower shape -
semidouble or double or in its
parts like petal or sepal or
serration is of high commercial
value
▪ Phenotype with unique flower
forms or, the double flower has
higher ornamental value than the
single
4/22/2021 Department of FLA
Figure 1: Forms of flower
5
6. Figure 2: Genetic analysis of flower development
4/22/2021 Department of FLA Suyama et al., 2010 6
7. ABC Flower Development Model
➢ George Haughn and Chris Somerville in 1988
➢ To explain how floral whorls develop
➢ Arabidopsis thaliana and Antirrhinum majus
4/22/2021 Department of FLA
Figure 3: ABC flower development model
Bowman et al. 1989 7
8. ABCDE Flower Development Model
➢ Arabidopsis thaliana and Snapdragon mutants
➢ Class A genes (APETALA1, APETALA2) controls sepal
development & together with class B genes, regulates the
formation of petals. Antirrhinum: LIPLESS 1 and 2
➢ Class B genes (e.g. PISTILLATA, and APETALA3), together
with class C genes, mediates stamen development. Antirrhinum:
DEFICIENS (DEF) and GLOBOSA (GLO)
➢ Class C genes (e.g., AGAMOUS), determines the formation of
carpel. Antirrhinum: PLENA (PLE)
➢ class D genes (e.g., SEEDSTICK, and SHATTERPROOF)
specify the identity of the ovule. Petunia: FBP7 and FBP11
➢ Class E genes (e.g., SEPALLATA), expressed in the entire floral
meristem and are necessary. (SEP1, SEP2, SEP3 and SEP4)
4/22/2021 Department of FLA Bowman et al. 1989 8
9. Figure 4: ABCDE flower development model
4/22/2021 Department of FLA
➢ Genes of ABCDE model are MADS-box genes.
Bowman et al. 1989 9
10. MADS-box
➢ Conserved sequence motif found in genes which comprise the MADS-box
gene family
➢ The MADS box encodes the DNA-binding MADS domain.
➢ MADS-domain proteins are generally transcription factors
➢ Length of the MADS-box - 168 to 180 base pairs
➢ Origin:
➢ MCM1 from the budding yeast, Saccharomyces cerevisiae
➢ AGAMOUS from the thale cress Arabidopsis thaliana
➢ DEFICIENS from the snapdragon Antirrhinum majus
➢ SRF (serum response factor) from the human Homo sapiens
➢ MADS-box genes - male and female gametophyte development, embryo and
seed development, root, flower and fruit development
➢ Floral homeotic MADS-box genes (AGAMOUS and DEFICIENS) – determine
floral organ identity according to the ABC model of flower development
4/22/2021 Department of FLA Bowman et al. 1989 10
11. HYBRIDIZATION
➢ Technique to induce variation in
floricultural crops
➢ Process of crossbreeding
between genetically dissimilar
parents to produce a hybrid.
➢ Crossing involves placing pollen
grains from one genotype, the
male parent, on to the stigma of
flowers of the other genotype,
the female parent
4/22/2021 Department of FLA
Figure 5: Hybridization technique
11
12. ➢ Unique shape is by crossing two different forms
➢ Single, double, semi-double depending upon genetic
constitutions
➢ Single, semi-double and double types of flower in ornamental
crops are genetically controlled either by a single gene or
multiple genes.
➢ Select suitable genotype as a parent to develop new cultivars
having required flower type
4/22/2021 Department of FLA
Gaurav et al. 2017
12
13. Table 1: Inheritance of double flower character in flower
crops
4/22/2021 Department of FLA Gaurav et al., 2017 13
15. MUTATION BREEDING
➢ Freisleben and Lein (1944)
➢ Mutation breeding
➢ An attractive method for creating genetic variability -
ornamental plant
➢ Ornamental plants - Ideal systems for mutagenesis
➢ Heterozygous and vegetatively propagated – detection,
selection and conservation – M1
➢ Mutation induction led to changes in floral morphology -
ornamental value
Schum and Preil, 1998
4/22/2021 Department of FLA 15
16. MUTAGENS
Physical mutagens
1. Ionizing radiation
(a)Particulate radiations:
alpha-rays, beta-rays, fast
neutrons and thermal
neutrons
(b) Non-particulate radiations:
X-rays and Gamma rays
2. Non-ionizing radiation:
ultraviolet radiation
Chemical mutagens
1. Alkylating agents: EMS,
MMS, sulphur mustard,
nitrogen mustard
2. Acridine dyes: Proflavin,
acridine orange, acridine
yellow and ethidium
bromide
3. Base Analogues: 5 Bromo
uracil, 5 chloro-uracil
4. De-amination agents: Nitrous
acid, Sodium azide
4/22/2021 Department of FLA Schum and Preil, 1998 16
17. ➢ Mutagenic treatment
➢ Acute – once in the shortest time
➢ Chronic – long period (weeks or months)
➢ Recurrent – two or more subsequent generations (M2, M3
or M4 after M1)
➢ Causes of mutation
➢ Genetic structure change
➢ Genome mutation
➢ Gene (point mutation) – alteration in nuclear DNA
➢ Specific sequence of nucleotide – new type of protein
4/22/2021 Department of FLA Schum and Preil, 1998 17
18. Figure 6: Types of point mutation
4/22/2021 Department of FLA Schum and Preil, 1998 18
19. Figure 7: Types of chromosomal mutation
4/22/2021 Department of FLA Schum and Preil, 1998 19
20. Euphytica, 2014, 199:317–324
Isolation of flower color and shape mutations by
gamma radiation of Chrysanthemum morifolium
Ramat cv. Youka
Tarek M. A., Soliman, Suhui Lv, Huifang Yang, Bo Hong, Nan Ma, Liangjun Zhao
College of Agronomy and Biotechnology, China
4/22/2021 Department of FLA 20
21. Materials and methods
➢ Location: College of Agronomy and Biotechnology, China Agricultural
University, Beijing, China
➢ Plant materials : C. morifolium Ramat cv. Youka (spoon shape)
➢ Explants - petals flower buds
➢ Sterilzation procedure - 75 % alcohol for 20–30 s and 0.1 % HgCl2 solution
➢ Buds were opened and the white petals 4 mm were excised and cultured on
MS basal medium (Murashige and Skoog)
➢ pH - 6.0
➢ Culture room - 16/8 h light/dark regimes at 26 ± 1 C, 2,000 lux provided by
cool white fluorescent tubes and 55–60 % relative humidity
➢ Callus - gamma radiation using 60Co of gamma chamber with doses of 0
(Control), 10, 15 and 20 Gy and dose rate 1.02 Gy/min.
➢ Three replications were applied to each dose and each replication consist of
100 explants.
4/22/2021 Department of FLA
Tarek et al., 2014 21
22. Figure 8: In vitro regeneration C. morifolium ‘Youka’ from ray florets. a) White flower
buds; b) Callus induction medium; c) Adventitious shoots formation after 4 weeks;
d) In vitro Roots formations after 25 days; e) Plantlets in hardening chamber
4/22/2021 Department of FLA Tarek et al., 2014 22
23. Table 2: In vitro callus survival (%) and number of shoots (Mean ± SE) of white
C. morifolium ‘‘Youka’’ as influenced by gamma ray doses
4/22/2021 Department of FLA
Gamma ray dose (Gy) Callus survival (%) No. of shoot
0 86.67 7.22 ± 0.11
10 62.43 7.67 ± 0.33
15 30.33 7.89 ± 0.29
20 17.23 3.00 ± 0.29
Tarek et al., 2014 23
24. 4/22/2021 Department of FLA
Table 3: Effect of in vitro treatment of C. morifolium ‘Youka’ with gamma
radiation on flowering characteristics of the generated plantlets
Character mean ±
SE
Treatments
Control 10 Gy 15 Gy 20 Gy
Flower no. per plant 4.22 ± 0.29 4.38 ± 0.12 4.05 ± 0.14 3.00 ± 0.00
Flower diameter
(cm)
6.12 ± 0.23 6.11 ± 0.15 5.15 ± 0.18 4.88 ± 0.06
Petal length (cm) 3.03 ± 0.20 3.14 ± 0.19 3.00 ± 0.01 3.01 ± 0.01
Petal width (cm) 0.83 ± 0.07 0.87 ± 0.12 0.09 ± 0.03 1.05 ± 0.06
Petiole length (cm) 4.50 ± 0.17 4.72 ± 0.18 6.52 ± 0.29 6.51 ± 0.30
Petiole diameter
(cm)
2.50 ± 0.11 1.89 ± 0.04 1.89 ± 0.04 1.85 ± 0.04
Tarek et al., 2014 24
25. Figure 9: Flower of tissue-raised plants of Chrysanthemum morifolium c.v Youka :
a. control, white colored with spoon shaped petals; b. M.1, white colored
with tubular petals; c. M.2, yellow colored with spoon shaped petals;
d. M.3, yellow colored with flat shaped petals
4/22/2021 Department of FLA Tarek et al., 2014 25
26. Figure 10: Flower shape mutants of carnation variety ‘Vital’ by ion
beam irradiation
4/22/2021 Department of FLA Okamura et al., 2003 26
27. Figure 11: Gamma irradiated Torenia hybrida flowers with erose petal
margins (control on the right, mutants treated with 0.0075
mM colchicine for 48 h and 30 Gy gamma radiation center
and left). Scale bar in cm.
4/22/2021 Department of FLA Suwansere et al., 2011 27
31. Table 4: Induced mutation for change in flower morphology in
flower crops
4/22/2021 Department of FLA 31
32. Limitation
➢ Random and unpredictable
➢ Frequency of desirable mutation is very low - 0.1% of the
total mutations.
➢ Useful mutants are rare and predominantly recessive
➢ Desirable mutation associate with undesirable side effects
➢ Mutants - strong negative pleiotropic effects on other traits
➢ Field trial and germplasm storage - expensive and require a
lot of space and careful management
➢ Screen large population to select desirable mutations
➢ Health risks: handling, chemical mutagens; radiations, fast
neutrons treatments
4/22/2021 Department of FLA 32
33. POLYPLOIDY
➢ Polyploidy – cells of an organism have more than two
paired sets of chromosome
➢ Doubling the chromosome number of a species
➢ Create variations in the species where the natural
variations are limited
➢ Genetic variations – breeding program
➢ Useful in several crops for breeding purpose
➢ Increased in size and shape of plants, their leaves,
branches, flower parts, fruits, and seeds
➢ Improved plant architecture - provide good material for the
breeding programme and for further development of
cultivars
Mata, 2009
4/22/2021 Department of FLA 33
34. Figure 13: Factors affecting plant in vitro artificial polyploidy induction system
4/22/2021 Department of FLA Niazian and Nalousi, 2020 34
35. COLCHICINE
➢ Alkoloid - Colchicum autumnale
➢ C22H25O6N
➢ Interferes with the development of spindle apparatus
➢ Sister chromatids of chromosomes are unable to migrate to the opposite poles –
anaphase
➢ Chromatids (=4) are included in the same restitution nucleus leading to
chromosome doubling
➢ Blakesle, Avery and Nebel in 1937.
4/22/2021 Department of FLA Mata, 2009 35
36. Figure 14: The phenomenon of giga after polyploidization in plants
4/22/2021 Niazian and Nalousi, 2020
Department of FLA 36
37. Folia Hort., 2018, 30(2): 307-319
Induction and identification of colchicine induced
polyploidy in Gladiolus grandiflorus ‘White
Prosperity’
Ayesha Manzoor, Touqeer Ahmad, Muhammad Ajmal
Department of Horticulture, PMAS Arid Agriculture University Rawalpindi,
Pakistan
4/22/2021 Department of FLA 37
38. MATERIAL AND METHODS
➢ Experimental site - Pakistan from September 2015 to April
2016.
➢ Plant material - Corms ‘White Prosperity - 2.6 cm
➢ The non-dormant corms were soaked in 0.1%, 0.2% and
0.3% colchicine solution, while control corms were soaked
in distilled water for 24 h
➢ Completely randomized design (CRD) having 4 treatments
and 3 replications consisting of 16 corms in each
replication.
Manzoor et al., 2018
4/22/2021 Department of FLA 38
39. Table 5: Reproductive parameters of control and colchicine treated plants
of gladiolus ‘White Prosperity’
Manzoor et al., 2018
4/22/2021 Department of FLA 39
40. Figure 15: Impact of different concentrations of colchicine on floret
diameter in gladiolus ‘White Prosperity’: floret diameter was
less in control (a) and at 0.1% (b) but it was increased at 0.2%
(c) and was maximum at 0.3%
Manzoor et al., 2018
4/22/2021 Department of FLA 40
41. Figure 16: Morphological variation in flower petals of gladiolus ‘White
Prosperity’: petals had smooth edges and triangular shape in control plants (a)
pointed outgrowth appeared on petal surface in treated plants along with
elongated petal shape and serrated margins at 0.1% colchicine (b), flower
produced oval shaped petals whose one end had ruffled edges at 0.2% colchicine
(c) petals with pointed outgrowth were also produced at 0.3% colchicine (d)
Manzoor et al., 2018
4/22/2021 Department of FLA 41
42. Figure 17: Flowers of the diploid (right) and tetraploid (left) regenerated
from sectioning of PLBs of P. schilleriana
4/22/2021 Department of FLA
Chen et al., 2008
42
43. GENETIC MODIFICATION
➢ Development of new ornamental varieties through gene
transfer
➢ Development of new varieties - hybridization or
mutagenesis is very difficult
➢ Introduce traits which can’t be generated by conventional
breeding.
➢ Major traits - flower color, fragrance, abiotic stress
resistance, disease resistance, pest resistance, manipulation
of the form and architecture of plants and/or flowers,
modification of flowering time, and post-harvest life etc.
4/22/2021 Department of FLA 43
44. RNAi (RNA INTERFERENCE)
➢ RNA interference (RNAi) is a sequence specific gene
silencing phenomenon caused by the presence of double
stranded RNA.
➢ Used as a knockdown technology.
➢ Analyze gene function in various organism
➢ RNAi targets include RNA from viruses and transposons.
4/22/2021 Department of FLA Singh et al. 44
46. Timeline
Anderw Fire Craig Mello
1968
Crick & Orgel proposed
that RNA was the first
information molecule.
1990
Jorgenson with petunia
discovered gene silencing
1990-92
The human genome was
initiated & sequence
information began to increase
exponentially
2009
Ghidiya et al., discovered
different types of silencing
RNA
2009-2011
Invention related to various
drug discovery & therapettics.
1998
Fire & Mello coined the term
RNAi for gene silencing
mechanism performed by
dsRNA molecules in C.
elegans
2001
Hammond’s group
discovered RISC in
Drosophila melanogaster
1995
Guo & Kemphues discovered
that either sense or antisense
RNA could lead to gene
silencing while working on C.
elegans
2006
Fire & Mello won Nobel Prize
for discovering RNAi
mechanism
1972
Noller proposed the role of rRNA
in translation of mRNA into protein
molecules.
http://www.rnaiweb.com/RNAi/RNAi_Timeline)
4/22/2021 Department of FLA 46
47. Other name of RNAi
➢Co-suppression
➢PTGS
➢Gene Silencing
➢Quelling
4/22/2021 Department of FLA 47
48. 4/22/2021 Department of FLA
Figure 18: RNAi Affecting Gene Expression
Bernstein et al., 2000 48
49. RNA
Coding RNA Non Coding
RNAs
mRNA
Constituent RNAs Regulatory RNAs
rRNAs
tRNAs
siRNAs miRNAs snRNAs
snoRNAs
Types of RNA involved in RNAi
4/22/2021 Department of FLA 49
50. COMPONENTS OF GENE SILENCING
• 1.Enzymes
• Dicer
• Drosha
• 2.RISC
• 3.RNA
• siRNA
• miRNA
• RdRp
4/22/2021 Department of FLA 50
51. DICER
➢ Endoribonuclease Dicer or helicase with
RNase motif
➢ Enzyme involve in the initiation of RNAi
➢ Rnase III family
➢ Recognise and cleaves double-stranded RNA
(dsRNA) and pre-microRNA (pre-miRNA)
into short double-stranded RNA fragments
called small interfering RNA and microRNA,
respectively.
➢ Dicer family proteins are ATP dependent
nucleases.
➢ Dicer facilitates the activation of the RNA-
induced silencing complex (RISC), which is
essential for RNA interference.
4/22/2021 Department of FLA
Bernstein et al., 2000 51
52. DICER’S DOMAINS
DICER’S DOMAINS
➢ Drosha :
➢ Core nuclease that executes the initiation step
of microRNA (miRNA) processing in
the nucleus
➢ Cleaves pri-miRNA, to form pre miRNA, which
is later processed by Dicer
Bernstein et al., 2000
Figure : One molecule of
the Dicer protein
from Giardia intestinalis;
RNase III domains are
colored green, the PAZ
domain yellow, the
platform domain red, and
the connector helix blue
➢ Both Drosha and Dicer can function as master
regulators of miRNA processing
4/22/2021 Department of FLA 52
53. RNA-induced silencing complex (RISC)
➢ Drosophila – Hammond
➢ Multi-protein complex –ribonucleoprotein
➢ Recognise and incorporate with one strand
of a single-stranded RNA (ssRNA)
fragment, such as microRNA (miRNA), or
double-stranded small interfering RNA
(siRNA)
➢ The single strand acts as a template for
RISC to
recognize complementary messenger RNA
(mRNA) transcript
➢ Activate Argonaute (a protein within
RISC) and cleave the mRNA.
4/22/2021 Department of FLA 53
54. Argonaute
➢ Active part of the RISC
➢ Binds different classes of small non-
coding RNAs, including miRNAs
and siRNAs and cleave the target
mRNA strand
➢ Endonuclease activity
➢ Responsible for selection of the
guide strand and destruction of the
passenger strand of the siRNA
substrate.
➢ Play a role in both triggering and
amplifying the silencing effect
4/22/2021 Department of FLA 54
55. RNA dependent RNA polymerase(RdRPs)
➢ RNA replicase
➢ Enzyme that catalyzes the replication of RNA from an RNA
template
➢ Transgenic plants show an accumulation of aberrant transgenic
RNAs, which is recognized by RdRPs and used as templates and
synthesize antisense RNAs to form dsRNAs
➢ dsRNAs formed are finally the targets for sequence specific
RNA degradation.
4/22/2021 Department of FLA Bernstein et al., 2000 55
56. miRNA (micro RNA)
➢ Non-coding RNA molecule
(containing about 22 nucleotides)
➢ RNA silencing and post-
transcriptional regulation of gene
expression
➢ miRNAs function via base-
pairing with complementary
sequences within mRNA molecules
➢ mRNA molecules are silenced
➢ (1) Cleavage of the mRNA strand
into two pieces, (2) Destabilization
of the mRNA through shortening of
its poly(A) tail, and
➢ (3) Less efficient translation of the
mRNA into proteins by ribosomes
4/22/2021 Department of FLA
Figure 19: miRNA biogenesis
Zhang et al., 2002 56
58. siRNA
➢ Small interfering - an integral role in the phenomenon of RNA
interference (RNAi), a form of post transcriptional gene silencing
➢ 21-25 nt fragments
➢ It interferes with the expression of specific gene
➢ Bind to the complementary portion of the target mRNA and tag
it for degradation preventing translation
➢ A single base pair difference between the siRNA template and
the target mRNA is enough to block the process.
4/22/2021 Department of FLA Zhang et al., 2002 58
60. Scientia Horticulturae, 2014, 178: 1–7
Double flower formation induced by silencing of C-
class MADS-box genes and its variation among
petunia Cultivars
Siti Hajar Noor, Koichiro Ushijima, Ayaka Murata, Kaori Yoshida, Miki
Tanabe, Tomoki Tanigawa, Yasutaka Kubo, Ryohei Nakano
Graduate School of Environmental and Life Science, Okayama University, Japan
4/22/2021 Department of FLA 60
61. MATERIALS AND METHODS
➢Plant materials - Petunia (P. hybrida) seeds of cultivars ‘Fantasy Blue’,
‘Pico-bella Blue’, ‘Cutie Blue’ and ‘Mambo Purple’
➢Plasmid construction - Tobacco rattle virus (TRV)-based VIGS system
➢- pTRV1 and pTRV2 VIGS vectors
➢A cDNA fragment of petunia chalcone synthase, PhCHS - pTRV2 vector
to form pTRV2 PhCHS
➢Petunia C-class genes, pMADS3 and FBP6 - SmaI site of pTRV2 PhCHS
vector individually to generate constructs for silencing pMADS3 and
FBP6 separately
➢Silencing pMADS3 and FBP6 simultaneously, pMADS3 and FBP6
fragments were fused and cloned into the SmaI site of pTRV2 PhCHS
vector.
4/22/2021 Department of FLA Noor et al., 2014 61
62. Figure 20: Morphological changes in flowers of P. hybrida cv ‘Cutie Blue’
inoculated with pTRV2-PhCHS/pMADS3 and pTRV2-PhCHS/pMADS3/FBP6 (a,
b) VIGS-untreated control flower, stamens and a carpel (c, d) pMADS3-VIGS
flower, petaloid stamens and a carpel (e, f) pMADS3/FBP6-VIGS flower, petaloid
stamens and a carpel Scale bars = 1 cm.
4/22/2021 Department of FLA Noor et al., 2014 62
63. Figure 21: Morphological changes in flowers of P. hybrida cv ‘Fantasy Blue’,
‘Picobella Blue’, and ‘Mambo Purple’ inoculated with pTRV2-
PhCHS/pMADS3/FBP6 (pMADS3/FBP6-VIGS). (a–c) ‘Fantasy Blue’; (d–f)
‘Picobella Blue’; (g–i) ‘Mambo Purple’; (a, d and g) VIGS-untreated control
flowers; (b, e and h) pMADS3/FBP6-VIGS flowers; (c, f and i) stamensand carpels
or converted new flowers of pMADS3/FBP6-VIGS flowers. Scale bars = 1 cm.
4/22/2021 Department of FLA Noor et al., 2014 63
64. Figure 22: New flower formation in whorl 4 and from axil of whorl 3 in a double
flower of P. hybrida cv ‘Mambo Purple’ inoculated with pTRV2-PhCHS/pMADS3/FBP6
(pMADS3/FBP6-VIGS). (a) An opened double flower with a second new flower in
whorl 4 and an ectopic new flower emerging from the axil of whorl 3; (b) an opened
second new flower;(c) fused corolla (left), a carpel (center), and petaloid stamens (right)
of the second flower; (d) an ectopic new flower emerging from the axil of whorl 3; (e) an
unconverted stamen (left) and petal-like tissues of the ectopic new flower.
4/22/2021 Department of FLA Noor et al., 2014 64
65. Figure 23: Average surface areas and average cell sizes of P. hybrida cv
‘Cutie Blue’ inoculated with pMADS3-VIGS) and
pMADS3/FBP6-VIGS
4/22/2021 Department of FLA Noor et al., 2014 65
66. Figure 24: Modified ray florets of a transgenic chrysanthemum with
reduced chrysanthemum-AGAMOUS gene expression. The pistil of each
ray floret was changed to several corolla-like tissues (secondary corolla)
and a pistil-like tissue.
4/22/2021 Department of FLA Aida et al., 2008 66
67. Figure 25: Flower phenotype of wild type and dp mutant
(Insertion of an En/Spm‐related transposable element into a floral homeotic
gene DUPLICATED causes a double flower phenotype in the Japanese morning
glory)
4/22/2021 Department of FLA Nitasaka et al., 2003 67
68. Figure 26: Transgenic gerbera with MADS box genes. (a) 35S-gaga2
plant. (b) 35S-antisense gglo1 plant
4/22/2021 Department of FLA Yu et al., 1999
➢ Ray florets in transformants with antisense gerbera AGAMOUS formed
corolla-like organs in the third whorl, and all floret types formed carpelloid-
and pappus-like organs in the fourth whorl; however, they maintained floral
determinacy
68
69. CRES-T
➢ Chimeric REpressor gene-Silencing Technology
➢ Unique gene-silencing method
➢ Recently developed for the functional analysis of plant
transcription factors and for the genetic manipulation of plant
traits.
➢ A transcription factor is converted to a strong chimeric repressor
by fusion with a transcriptional repression domain, SRDX
➢ The chimeric repressor dominantly represses the expression of
target genes, even in the presence of redundant endogenous
transcription factors
➢ Result in a loss-of-function phenotype of the transcription factor
4/22/2021 Department of FLA Narumi et al., 2011 69
70. Plant Biotechnology, 2011, 28: 131–140
Arabidopsis chimeric TCP3 repressor produces novel
floral traits in Torenia fournieri and Chrysanthemum
morifolium
Takako Narumi, Ryutaro Aida, Tomotsugu Koyama, Hiroyasu Yamaguchi,
Katsutomo Sasaki, Masahito Shikata, Masayoshi Nakayama, Masaru
Ohme-Takagi2, Norihiro Ohtsubo
National Institute of Floricultural Science, Tsukuba, Ibaraki, Japan
4/22/2021 Department of FLA 70
71. MATERIALS AND METHODS
➢ Plant material - Torenia fournieri Lind. ‘Crown Violet’ and
Chrysanthemum morifolium Ramat. ‘Sei-Marin’ (Seikoen, Hiroshima,
Japan)
➢ Generation of transgenic plants - TCP3-SRDX, TCP3-mSRDX, and
TCP3-ox genes - introduce into the destination vector pBCKK
➢ Each transgene vector introduce into Torenia and chrysanthemum by
Agrobacterium-mediated transformation
➢ Transgenic plants regenerated - grown in the contained greenhouse
under natural light.
➢ Anatomical observation of floral organs - Fresh tissues were prepared
after the opening of flower buds and examined by scanning electron
microscope without fixing in order to observe petal and stigma
surfaces.
4/22/2021 Department of FLA Narumi et al., 2011 71
72. Figure 27: Phenotypic comparison of torenia flowers and leaves.
Narumi et al., 2011
4/22/2021 Department of FLA 72
73. 4/22/2021 Department of FLA
Figure 28: Comparison of petal color and shape in transgenic torenia
plants.
Narumi et al., 2011 73
74. Figure 29: Phenotypic comparisons of chrysanthemum flowers and leaves.
Narumi et al., 2011
4/22/2021 Department of FLA 74
75. 4/22/2021 Department of FLA
Figure 30: Comparison of corolla size, petal shape, and disk floret maturity
in transgenic chrysanthemum plants. . Bars = 1 cm.
Narumi et al., 2011 75
76. Figure 31: Morphological changes in the epidermal cells of TCP3-SRDX torenia petals
4/22/2021 Department of FLA Narumi et al., 2011 76
77. Figure 32: Expression of the TCP3-SRDX transgene in torenia and chrysanthemum. (A)
Total RNAs were prepared from petals of wild-type and TCP3-SRDX torenias
with type I, type II and type III phenotypes. (B) Total RNAs were prepared
from the leaves of wild-type and TCP3-SRDX chrysanthemums with type I and
type II phenotypes.
4/22/2021 Department of FLA Narumi et al., 2011 77
78. CONCLUSION
➢ Suppress endogenous TCP related activities in torenia and
chrysanthemum, without isolating and identifying their
sequences or target genes.
➢ TCP3-SRDX - chimeric repressors of development- and
differentiation-related transcription factors may have the ability
to modify petal colors and patterns in addition to petal size and
shape
➢ Modify cellular identity or positional information within the
petal.
4/22/2021 Department of FLA 78
79. Gene transfer methods
Indirect
➢ Agrobacterium
mediated gene transfer
➢ Most widely used
➢ More economical
➢ More efficient
➢ Transformation success
is 80-85%
Direct
➢ Particle bombardment
or micro projectile
➢ Direct DNA delivery
by Microinjection
➢ Ultrasonication
➢ Electroporation
4/22/2021 Chandler and Brugliera, 2011
Department of FLA 79
81. Future thrust
➢ Identification of flower shape mutant and development of
in-vitro protocol for their regeneration
➢ Most of the genetic modification for flower form is done in
Arabidopsis & Snapdragon only, it has be done in
commercially important crop
➢ Only few GM crop is released for commercial purpose,
i.e., Rose & Carnation (flower colour). Effort is to be done
to developed GM crop with modified flower architecture
4/22/2021 Department of FLA 81
82. CONCLUSION
➢ Creation of new flower shapes in ornamental plants is a
major breeding target as increase its commercial value.
➢ Flower shape including double flower can be developed by
different breeding techniques - hybridization, mutation,
polyploidy
➢ RNAi and CRES-T
4/22/2021 Department of FLA 82