This document summarizes genetic engineering techniques used in floriculture to modify flower traits. It discusses how genes involved in pigment synthesis can be overexpressed or silenced to modify flower color. For example, introducing the F3'5'H gene produces blue carnations. It also describes how traits like vase life and disease resistance can be improved. Overall, the document outlines various genetic engineering approaches that have been applied to modify important commercial flower qualities.
seminar 1 on FLOWER COLOUR MODIFICATION by SK SAMIM AHAMMED, M.Sc(Ag), genetics and plant breeding, BIDHAN CHANDRA KRISHI VISWAVIDYALAYA, MOHANPUR, NADIA,WEST BENGAL,INDIA.
The presentation gives overview of production of secondary metabolites using callus culture as well as tissue culture techniques. Various batch and continuous culturing process are described on the basis of secondary metabolite to be synthesised.
Somaclonal Variation in Plant tissue culture - Variation in somaclones (somatic cells of plants)
Somaclonal variation # Basis of somaclonal variation # General feature of Somaclonal variations # Types and causes of somaclonal variation # Isolation procedure of somaclones via without in-vitro method and with in-vitro method with their limitations and advantages # Detection of isolated somaclonal variation # Application (with examples respectively related to crop improvement) # Advantages and disadvantages of somaclonal variations.
https://www.youtube.com/watch?v=IZwrkgADM3I
Also watch, Gametoclonal variation slides to understand, how to changes occur in gametoclones of plants.
https://www.slideshare.net/SharmasClasses/gametoclonal-variation
Meristem tip culture for the production of the virus free plantsArjun Rayamajhi
This presentation gives general idea on the meristem tip culture for the production of the virus free plants. The principles, methods and procedures of the meristem tip culture included. General idea on different in vitro culture techniques for virus elimination meristem tip culture viz. thermotherapy, cryotherapy,chemotherapy and electrotherapy are provided.
seminar 1 on FLOWER COLOUR MODIFICATION by SK SAMIM AHAMMED, M.Sc(Ag), genetics and plant breeding, BIDHAN CHANDRA KRISHI VISWAVIDYALAYA, MOHANPUR, NADIA,WEST BENGAL,INDIA.
The presentation gives overview of production of secondary metabolites using callus culture as well as tissue culture techniques. Various batch and continuous culturing process are described on the basis of secondary metabolite to be synthesised.
Somaclonal Variation in Plant tissue culture - Variation in somaclones (somatic cells of plants)
Somaclonal variation # Basis of somaclonal variation # General feature of Somaclonal variations # Types and causes of somaclonal variation # Isolation procedure of somaclones via without in-vitro method and with in-vitro method with their limitations and advantages # Detection of isolated somaclonal variation # Application (with examples respectively related to crop improvement) # Advantages and disadvantages of somaclonal variations.
https://www.youtube.com/watch?v=IZwrkgADM3I
Also watch, Gametoclonal variation slides to understand, how to changes occur in gametoclones of plants.
https://www.slideshare.net/SharmasClasses/gametoclonal-variation
Meristem tip culture for the production of the virus free plantsArjun Rayamajhi
This presentation gives general idea on the meristem tip culture for the production of the virus free plants. The principles, methods and procedures of the meristem tip culture included. General idea on different in vitro culture techniques for virus elimination meristem tip culture viz. thermotherapy, cryotherapy,chemotherapy and electrotherapy are provided.
Present status of Floriculture industry in Tamilnadu, India & World. All other aspects of floriculture industry including merits, demerits, oppurtunities, strategies to develop, potentials, scope etc..
Recent advances in neutraceutical and value addition in ornamental horticulture Aparna Veluru
nutraceuticles, edible flowers, flowers used as a medicine and different ornamental plants with medicinal importance, value addition of ornamental plants are mentioned
Plant phenolics are secondary metabolites that encompass several classes structurally diverse of natural products biogenetically arising from the shikimate-phenylpropanoids-flavonoids pathways. Plants need phenolic compounds for pigmentation, growth, reproduction, resistance to pathogens and for many other functions. Therefore, they represent adaptive characters that have been subjected to natural selection during evolution. Plants synthesize a greater array of secondary compounds than animals because they cannot rely on physical mobility to escape their predators and have therefore evolved a chemical defence against such predators. This article, after a short review of plant phenols and polyphenols as UV sunscreens, signal compounds, pigments, internal physiological regulators or chemical messengers, examines some findings in chemical ecology concerning the role of phenolics in the resistance mechanisms of plants against fungal pathogens and phytophagous insects.
Role of biotechnology in enhancing fruit crop production and qualityankit gawri
It was evident that developed biotechnological approaches have the potential to enhance the yield, quality, and shelf-life of fruits and vegetables to meet the demands of the 21st century. However, the developed biotech approaches for fruits and vegetables were more of academic jargon than a commercial reality
Phytochemical Potential and Antimicrobial Activity of Andrographispaniculataiosrjce
The Herbal medicine today ensures safety in contrast to the synthetic preparations. Herbs the Nature’s
Physician, have been reported as an important source of medicine for years and years. Using of herbs for
curing diseases dated back to prehistory and people of all continents have this old tradition.Recently, wide
research proposals highlight the property of medico potential from phytalsources. My herb of interest is also the
above said, ofcourseAndrographispaniculata (Acanthaceae) is a medicinal plant used for the treatment of
various ailments, which has been documented in history of all civilizations. The present study is to learn the
phytochemical properties and the antimicrobial activity of the above using disc diffusion method
Plant pigments that are responsible for their yellow, bright red, and orange color are carotenoids that play a key role in plant health. Therefore, consumption of foods that have carotenoids provide various health benefits to the human body. Different vegetables and fruits in which carotenoids are present yams, carrots, sweet potatoes, watermelon, papaya, cantaloupe, spinach, mangos, kale, tomatoes, oranges, bell peppers, etc. There are over 600 known carotenoids in nature, found in plant cells, bacteria, and algae, and that belongs to a class of phytonutrients “plant chemicals” . Xanthophyll is the molecules known as hydrocarbons and found in orange and yellow fruits and vegetables, for example, pumpkin, cantaloupe, sweet potatoes, apricots and carrots. Association of word carrot with carotene will certainly relate the color of these pigments. Later class is oxygen containing molecules, found in dark leafy greens such as kale, spinach, and broccoli.Plant pigments, a generic term used to designate a large number of colored molecules, can be classified into tetrapyrroles e.g., chlorophyll and carotenoids e.g., ß carotene and xanthophyll . Of the pigments, carotenoids are fat soluble lipophilic natural pigments which are synthesized by plants and some microbes. These pigments not only play a key role in photosynthesis but are also responsible for the bright colors of various plants, fruits, flowers, and vegetables. Sheikh Ayyan Ahad "Plant Pigments as Drugs" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-6 | Issue-6 , October 2022, URL: https://www.ijtsrd.com/papers/ijtsrd52160.pdf Paper URL: https://www.ijtsrd.com/chemistry/other/52160/plant-pigments-as-drugs/sheikh-ayyan-ahad
Antimicrobial Drug Synthesis from Submerge Cultures of Pleurotus florida in D...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
hematic appreciation test is a psychological assessment tool used to measure an individual's appreciation and understanding of specific themes or topics. This test helps to evaluate an individual's ability to connect different ideas and concepts within a given theme, as well as their overall comprehension and interpretation skills. The results of the test can provide valuable insights into an individual's cognitive abilities, creativity, and critical thinking skills
The ability to recreate computational results with minimal effort and actionable metrics provides a solid foundation for scientific research and software development. When people can replicate an analysis at the touch of a button using open-source software, open data, and methods to assess and compare proposals, it significantly eases verification of results, engagement with a diverse range of contributors, and progress. However, we have yet to fully achieve this; there are still many sociotechnical frictions.
Inspired by David Donoho's vision, this talk aims to revisit the three crucial pillars of frictionless reproducibility (data sharing, code sharing, and competitive challenges) with the perspective of deep software variability.
Our observation is that multiple layers — hardware, operating systems, third-party libraries, software versions, input data, compile-time options, and parameters — are subject to variability that exacerbates frictions but is also essential for achieving robust, generalizable results and fostering innovation. I will first review the literature, providing evidence of how the complex variability interactions across these layers affect qualitative and quantitative software properties, thereby complicating the reproduction and replication of scientific studies in various fields.
I will then present some software engineering and AI techniques that can support the strategic exploration of variability spaces. These include the use of abstractions and models (e.g., feature models), sampling strategies (e.g., uniform, random), cost-effective measurements (e.g., incremental build of software configurations), and dimensionality reduction methods (e.g., transfer learning, feature selection, software debloating).
I will finally argue that deep variability is both the problem and solution of frictionless reproducibility, calling the software science community to develop new methods and tools to manage variability and foster reproducibility in software systems.
Exposé invité Journées Nationales du GDR GPL 2024
Richard's aventures in two entangled wonderlandsRichard 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.
ESR spectroscopy in liquid food and beverages.pptxPRIYANKA PATEL
With increasing population, people need to rely on packaged food stuffs. Packaging of food materials requires the preservation of food. There are various methods for the treatment of food to preserve them and irradiation treatment of food is one of them. It is the most common and the most harmless method for the food preservation as it does not alter the necessary micronutrients of food materials. Although irradiated food doesn’t cause any harm to the human health but still the quality assessment of food is required to provide consumers with necessary information about the food. ESR spectroscopy is the most sophisticated way to investigate the quality of the food and the free radicals induced during the processing of the food. ESR spin trapping technique is useful for the detection of highly unstable radicals in the food. The antioxidant capability of liquid food and beverages in mainly performed by spin trapping technique.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
ANAMOLOUS SECONDARY GROWTH IN DICOT ROOTS.pptxRASHMI M G
Abnormal or anomalous secondary growth in plants. It defines secondary growth as an increase in plant girth due to vascular cambium or cork cambium. Anomalous secondary growth does not follow the normal pattern of a single vascular cambium producing xylem internally and phloem externally.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Toxic effects of heavy metals : Lead and Arsenicsanjana502982
Heavy metals are naturally occuring metallic chemical elements that have relatively high density, and are toxic at even low concentrations. All toxic metals are termed as heavy metals irrespective of their atomic mass and density, eg. arsenic, lead, mercury, cadmium, thallium, chromium, etc.
Travis Hills' Endeavors in Minnesota: Fostering Environmental and Economic Pr...Travis Hills MN
Travis Hills of Minnesota developed a method to convert waste into high-value dry fertilizer, significantly enriching soil quality. By providing farmers with a valuable resource derived from waste, Travis Hills helps enhance farm profitability while promoting environmental stewardship. Travis Hills' sustainable practices lead to cost savings and increased revenue for farmers by improving resource efficiency and reducing waste.
Travis Hills' Endeavors in Minnesota: Fostering Environmental and Economic Pr...
Genetic engineering in floriculture
1. GENETIC ENGINEERING IN FLORICULTURE
Speaker : Zaman Mariya S.
Course No. : MBB 591
Major Sub. : Plant Biotechnology
Reg. No. : 04-1273-2010
ANAND AGRICULTURAL UNIVERSITY
Major Guide : Dr. G. C. Jadeja
Minor Guide : Dr. J. G. Talati
Date :15 /09/2012
Time : 14:00 Hrs
wel come
2. The Floriculture Industry……….
Biotechnology In Floriculture
Current Status Of Genetic Modification In
Floriculture
Transformation
Color Modification
Vase life
Flower Scent
Modified Plant Structure and Architecture
Disease Resistance
Case studies
Conclusion
Future Prospects
2
3. THE FLORICULTURE INDUSTRY
• Floriculture is considered to include the cut flowers, potted
plants, and ornamental bedding plants and garden plant
industries (Chandler, 2003).
• The worldwide production value of flowers and floricultural
plants is approx. 50 billion EURO.
• From the per capita consumption data provided by the Dutch
flower council (Flower Council of Holland, 2007), it can be
extrapolated that the global consumption value at consumer
level is somewhere between 100 to 150 billion EURO.
• The main areas of production and consumption of floricultural
products are in the United States and Europe, with a
significant industry in Japan. The area of production in China
is also increasing rapidly.
• The highest consumption per head is in the Netherlands,
Luxembourg, Germany, Austria, and France.
3
5. World Flower Production and Consumption
Production Consumption
China
Kenya
Columbia
Japan
Europe
USA
$40 billion
(Getu, 2009)
Market grow
20% annually
Japan
EU
USA
5
6. The global flower industry thrives on novelty.
Genetic engineering is providing a valuable means of
expanding the floriculture gene pool so promoting the
generation of new commercial varieties.
Engineered traits are valuable to either the consumer or
the producer.
At present, only consumer traits appear to provide a
return capable of supporting what is still a relatively
expensive molecular breeding tool.
The goal of genetic engineering is to improve the
characteristics of flowers such as, flower colour, vase life,
floral scent, flower morphology, disease as well as pest
resistance, flower productivity, timing and synchrony of
flowering.
Biotechnology In Floriculture
6
8. Genetic modification can be used to transfer
new
specific traits into the plant
Conventional Breeding
many gene and limited by genetic
incompability
Plant biotechnology
single gene with no specific to plant
species
Genetic engineering: Manipulation of plant genome
through recombinant DNA technology to alter plant
characteristics.
8
9. Gene transfer methods
Indirect Direct
Most widely used
More economical
More efficient
Transformation success is 80-85%
Agrobacterium mediated
gene transfer
Particle bombardment or
micro projectile
Direct DNA delivery by
Microinjection or
PEG mediated uptake
Ultrasonication
Electroporation
Chandler and Brugliera, 2011
9
10. Gene transformation
Bacterium mixed
with plant cells
GENE
identified and
isolated
Gene inserted
into Ti plasmid
Gene replication
Gold particles
coated with
DNA
Cells shot with gene gun and
DNA incorporated into plant cell
chromosome
Ti plasmid moves
into plant cell and
inserts DNA into
plant chromosome
Cells screened
for transgene
Transformed cells
selected with
selectable marker
Transgenic plant
regenerated from single
transformed cell 10
11. Color and color patterns
Flower color: most important trait, dictating consumer
attraction
Role of color:
• Attraction of pollinators
• Function in photosynthesis
• In human health as antioxidants and precursors of
vitamin A
• Protecting tissue against photooxidative damage
• Resistant to biotic and abiotic stress
• Symbiotic plant-microbe interaction
• Act as intermediary for other compounds
Color pattern: Differential accumulation of pigment(s)
11
14. Factors on flower color perception
pH of the vacuole:
pH of the vacuole is acidic
Small changes of pH have visible effects on flower color
Metal ions:
Metal complexing has a blueing effect on flower color
Co-pigmentation:
Co-pigmentation of anthocyanins with the colourless flavonols and
flavones is an important factor influencing pigmentation.
Co-pigmented flowers give a mauve colour, whereas in the absence
of flavonols maroon flowers are formed.
Co-occurrence of anthocyanins and yellow pigments
Mixtures of Ans and yellow flavonoids were found in the orange-
yellow or orange-red flowers of antirrhinum and bronze flower colour
of helichrysum.
Tanaka et al., 2009
low pH to high pH
15. Protein (Enzyme)
Red Pigment
Springob et al., 2003
Genes contain regulatory region
and coding region
Regulatory region Coding region
The genes consist of DNA which is
made up of four chemical letters
The chromosome is
made up of genes
Chromosomes-23 pairs
A cell
15
16. Genes involved in pigment synthesis
Structural (enzyme) gene: Gene that codes for any
RNA or protein product other than a regulatory protein.
Enzyme Gen
e
Species
CHS (Chalcone synthase) Chs Antirrhinum, Chrysanthemum, Orchid, Rosa,
Dianthus
CHI (Chalcone-flavanone
isomerase)
Chi Antirrhinum, Petunia, Eustoma, Dianthus
F3H (Flavone 3-hydroxylase) F3h Antirrhinum, Calistephus, Chrysanthemum,
Dianthus, Orchid
F3’H (Flavonoid 3’ hydroxlase) F3’h Antirrhinum, Dianthus, Petunia
F3’5’H (Flavonoid 3’,5’-
hydroxlase)
F3’5’h Calistephus, Eustoma, Petunia
FLS (Flavonol synthase) Fls Petunia, Rosa
FNS (Flavone synthase) FnsII Antirrhinum, Gerbera
DFR (Dihydroflavonol-4-
reductase)
Dfr Antirrhinum, Calistephus, Gerbera, Orchid,
Dianthus, Petunia Vainstein, 2004
Regulatory Genes
Structural (Enzyme) Genes
17. Regulatory genes: Influence the type, intensity and pattern of
flavonoid
accumulation but do not encode flavonoid enzyme.
Two classes of regulatory genes are
identified:
TF with MYB domain
TF with MYC/bHLH motif
(Vainstein, 2004)
Plant Gene
Myb Myc
Petunia Rosea, mixta Delia
Gerbera Gmyc I
Perilla MybpI
Petunia Phmyb3, An2,
An4
An1
17
20. Colour modification done by:
Over expression of structural genes
Use of sense or antisense enzyme construct
Inhibition of key biosynthetic enzyme
Addition of an enzyme in a particular biosynthetic step
20
21. FLORIGENE scientists Isolated the blue gene in 1991 and
patented in 1992.
Petunia gene didn’t work in roses, so FLORIGENE
scientists used their techniques on carnations— a much
easier species to manipulate than roses.
In 1996, Florigene developed mauve-coloured
carnation, FLORIGENE Moondust and it was the
world's first genetically modified flower on sale.
In 1997, developed second genetically-modified
carnation, FLORIGENE Moonshadow with a richer and
true purple colour.
Successfully developed a range of transgenic violet
carnations by introduction of a F3′5′H gene together
with a petunia DFR gene into a DFR-deficient white
carnation.
Fukui et al., 2003
mooniquemoonpearlmoonvelvetmoonberry
21
22. • Why a natural rose could not have the true blue colour?
"Flavonoid 3', 5'- hydroxylase" is one of the key enzymes
involved in the flavonoid biosynthesis for blue colour
development deficient in rose.
Natural rose did not have delphinidin.
pH of cell sap is 4.0-4.5 (acidic).
Cell sap is govern by 7 genes and each gene contributes 0.5
pH Anthocyanidin
The price for a single blue rose is
about $22 to $33. Source: The
Japan Today
The transgenic rose variety
‘‘Applause’’ was commercially
released in Japan in 2009 (Tanaka
et al., 2009)
22
23. Blue Gene Technology
www.suntory.com and www.florigene.com.au
In April of 2005, Suntory Ltd. and Florigene Ltd. announced the
production of a blue rose by introducing three transformation constructs
simultaneously into roses:
To make a blue
roses:
(1) Turn off’ the
rose DFR
gene
(2) Insert pansy
gene to open the
‘blue’ door
(3) Insert iris DFR gene
to make blue
pigment
23
24. Silencing anthocyanin biosynthetic pathway by
a) Transcriptional down-regulation
b) By inactivating the key enzymes
CHS
gene
F3’5’H
gene
DFR
gene
More sensitive to ranges of stresses
Alternative targets, better
and increased fragrance
Zuker et al., 200224
25. Gerbera
(Courtney-Gutterson et al. 1994),
Genetic engineering for white flower
(Elomaa 1993),
ChrysanthemumPetunia
(Krol et al. 1988),
Rose
(Gutterson 1995),
Carnation
(Gutterson 1995).
25
26. Genetic engineering for red/
orange flowers
•Cyanidin and delphinidin derivatives occur
but no pelargonidin derivatives in petunia.
•Petunia DFR cannot able to reduce
dihydrokaempferol because it shows
substrate specificity which lead to the
synthesis of leucoanthocyanidins.
•Over expression of A1 gene + abundant
substrate –opens a new pathway leading
to the synthesis of novel brick red colour
petunia.
Maize A1 gene
encodes
dihydroquercetin
4 reductase
Meyer et al., 198726
27. Genetic engineering for yellow
flowers
• Aurones are bright yellow
flavonoids found in species
such as snapdragon, dahlia
etc..
• Aurone synthase, more specifically
aureusidin synthase (AS), was
purified from yellow snapdragon
petals and the cDNA encoding the
enzyme was cloned.
• A pale yellow petunia expressing a
Lotus japonica PKR gene.
27
•Chalcone and aurones contribute to yellow
colours
• The most common chalcone, THC, is
yellow but is spontaneously
isomerized to naringenin in vitro and
rapidly isomerized in vivo by CHI.
• Discovery of chalcone 2′-
glucosyltransferase (C2′GT)
enzyme - stabilizes the
chemically unstable chalcone.
• Recent activity: (Okuhara et al.
2004)
Lack of CHI activity + presence
of UDP-glucose: THC 2’
glucosyltransferase (C2’GT)
activity = yellow carnation
28. Plant species Original
colours
Gene sources Methods Produced
flower colors
References
Cyclamen persicum Purple Endogenous F3’5’H Antisense Red to pink Boase et al. (2010)
Gentiana sp. Blue Endogenous CHS Antisense White Nishihara et al.
(2006)
Blue Endogenous F3’5’H RNAi Lilac to purple blue Nakatsuka et al.
(2008b)
Nierembergia sp. Violet Endogenous F3’5’H Antisense Pale blue Uyema et al. (20006)
Ostespermum
hybrida
Magenta Endogenous F3’5’H RNAi reddish Seitz et al. (2007)
Gerbera hybrida DFR Over expression
Petunia hybrida Blue Mazus psonilum CHS Dominant negative Pale blue Hanumappa et al.
(2007)
Red Lotus japonicus PKR Over expression Variegated red Shimada et al. (2006)
Rosa hybrida Red to pale cyanic Viola sp. F3’5’H Over expression Bluish Katsumoto et al.
(2007)
Torenia hybrida Blue Endogenous ANS RNAi Whitish to pale blue Nakamura et al.
(2006)
Blue Endogenous ANS Antisense Pale blue Nakamura et al.
(2006)
Blue Antirrhinum majus
AS
Over expression yellow Ono et al. (2006)
Flower colour modifications by regulating flavonoid biosynthesis
28
29. value of a cut flower =Post harvest longevity
Senescence highly controlled process requires active gene
expression & protein synthesis for programmed cell death.
Increased respiration and ethylene production induction of
catabolic enzymes resulting in decreased proteins
Genetic engineering for
longer vase life
Aging of
petals
Inhibiting or
by blocking
perception
of ethylene
29
30. Florigene has developed carnation flowers with
enhanced vase life using antisense RNA technology.
Down regulated
petal ACC
synthase
Control STS Transgenic
30
31. A high level of resistance in osmotin, pr-1 and/or
chitinase genes to a major carnation pathogen
(fusarium oxysporum f. Sp. Dianthi ) was
demonstrated in greenhouse tests. (Zuker et al.,
2005)
Genetic Engineering for biotic stress
Transformation of chrysanthemum cultivar 'shuho-
no-chikara' was modified by delta-endotoxin gene
cry1ab (mcbt) from bacillus thuringiensis
biological activity against lepidopteran insects into
chrysanthemum.
Protection of flower crops against coat protein
viruses (William R. Woodson 1991)
Transgenic chrysanthemum showing resistance
against chrysanthemum stunt viroid (csvd) and
TSWV.
31
32. Genetic engineering for improved shape,
size
ABC MODEL
32
The combination of three genes that
give rise to the flower parts.
A sepals
A + B petals
B + C stamens
C carpels
• The ABC model (Coen and
Meyerowitz 1991) and its modified
version (Theißen 2001) are known to
be applicable to a broad range of
plants (Kim et al. 2005).
33. 33
Genetic engineering for improved shape,
size
Constitutive expression of Antirrhinum majus
B genes DEF And GLO in transgenic torenia
resulted in the conversion of sepals to petals.
Constitutive expression of the C gene from
Rosa rugosa in torenia resulted in a carpeloid
structure in place of sepals (Kitahara et al.
2004)
Studies on homeotic mutants have clarified
many important aspects of genetic control on
flower development.
Deficiencies genes and AGAMOUS genes
isolated from Antirrhinum majus increased
interest in novel flower shapes through
molecular manipulation.
Transformant Wild type
34. Genetic engineering
for floral scent
may enhance the value of cut flowers to consumers…
Fragrance numerous
volatile aromatic organic
substances present in the
flower.
Such as,
hydrocarbons,
alcohols,
aldehydes,
ketones, esters,
ethers
Manipulation of fragrance in
flowers chemicals contributing to
the fragrance of roses, their
pathways of synthesis and
enzymes controlling these
pathways to be identified.34
35. Genetic engineering to modify
plant architecture
Control of plant height is of great importance in floriculture
35
• A petunia plant transformed with
Arabidopsis gai-1 (right).
• Genetic modification may replace
chemical growth retardants in
future.
36. Limitation of Plant gene transformation
• Improvement of traits controlled by many gene.
• Improvement of traits controlled by gene that has not
been identified.
• Requires high technology knowledge and equipment.
• Expensive cost.
• Government regulation.
36
37. o Flower colour changed from purple to almost white by the
down-regulation of the CHS gene
Surfinia Purple Mini
Tsuda et al., 2004
Surfinia Pure White
37Japan
1
Case studies
38. Flowers of transgenic Surfinia Purple Mini plant harboring
antisense DFR gene
Expression of DFR gene change the expression of the flavonol
synthase and flavone synthase gene
Contd…..
C
38Japan Tsuda et al., 2004
39. Vector construction
Figure 2. Schematic representation of the expression cassettes in T-DNA
of binary vectors constructed for colour modification.
39Japan
40. Generation of orange petunia expression of rose DFR gene
+ Suppression of F3H gene by antisense or RNAi method
Generation of red petunia expression of rose DFR gene
T-DNA copy number analysis of transgenic plants show that
three most stable lines, two plants had a single copy insert,
and the other one had two.
Contd…..
40Japan Tsuda et al., 2004
41. Modification of co-pigment by Suppressing flavonol biosynthesis resulted in
darker and slightly redder colour.
A flower of a transgenic plant expressing rose FLS gene has a paler flower
Surfinia Violet Mini
transgenic petunia
expressing torenia
FNSII gene
Creeping character did not change
41Japan Tsuda et al., 2004
42. Engineering of the Rose Flavonoid Biosynthetic
Pathway Successfully Generated Blue-Hued Flowers
Accumulating Delphinidin
Katsumoto et. al (2007)Japan
o Rosa hybrida lacks violet to blue colour due to the absence of
flavonoid 3’,5’-hydoxylase (F3’5’H) enzyme which produces
delphinidin-based anthocyanins.
o Other factors such as co-pigments and vacuolar pH also affect
flower colour.
o Expression of the viola F3’5’H gene accumulates (~95% high)
delphinidin a novel bluish flower colour.
42
2
43. • For more exclusive and dominant accumulation of
delphinidin irrespective of the hosts, the endogenous
dihydroflavonol 4-reductase (DFR) gene was down-
regulated and overexpressed the Iris3hollandica DFR
gene in addition to the viola F3’5’H gene in a rose
cultivar.
• The resultant roses exclusively accumulated delphinidin
in the petals, and the flowers had blue hues not achieved
by hybridization breeding.
• Moreover, the ability for exclusive accumulation of
delphinidin was inherited by the next generations.
43 Katsumoto et. al (2007)Japan
45. WKS77 WKS82 WKS100
WKS116 WKS124 WKS140
Rose Varieties transformed with pSPB130 and their flower colour changed are shown
Schematic representation of T-DNA region of binary vectors constructed for colour
modification for the constitutive over expression of the viola F3’5’H BP40 gene
and the torenia 5AT gene in rose.
45
46. o Correlation of delphinidin content and petal colours in transgenic Lavande.
o Pure red and blue have hue values of 360° and 290° in the hue angle.
46
Japan Katsumoto et. al (2007)
47. Host flower
violet-coloured
transgenic flower
98% delphinidin
98% delphinidin Madam Violet Seiryu
The vector pSPB919 is to down-regulate the endogenous rose DFR gene using RNA
interference (RNAi) and to over express the iris DFR and the viola F3’5’H genes.
48. • Northern blot analysis of LA/919-4-10.
• The expected sizes of the transcripts of
viola F3’5’H BP40 (1.8 kb) and iris DFR (1.7
kb) genes & smaller size was detected for
rose DFR mRNA (A).
• A rose DFR probe detected about 23 bp
small sized RNA, which was supposed to be
a degraded endogenous rose DFR transcript
with RNAi (B).
• Delphinidin contents was confirmed
in all transgenic (KmR) progeny of
LA/919-4-10.
• The flowers of F1 and F2 progeny
contained exclusively delphinidin.
48
(si RNA)
50. •Expressions of anthocyanin
biosynthetic genes in
transgenic gentian plants.
•The membranes were
hybridised with DIG-labelled
probes for GtCHS, GtF3’5’H
and 5/3’AT, respectively.
•WT indicates untransformed
wild-type gentian cv. ‘Albireo’.
Ethidium bromide-stained
ribosomal RNA bands (rRNA)
are shown as a control.
•Accumulation of anthocyanidin and flavone derivatives
in transgenic gentian plants.
(B) Flavone conc. in the petal
of an untransformed
control plant (WT) and
transgenic plants were
determined by measuring
the absorbance at 330 nm
using HPLC analysis with
apigenin and luteolin as
the standards.
• Data are the average ±SD
of five replicate flowers.
(A) Anthocyanidin conc. in the
petal of an untransformed
control (WT) and transgenic
plants were determined.
• Data are the average ±SD of
five replicate flowers.
50
RNA gel blot analysis
51. Mercuri et. al (2001)Italy
GFP detection in Eustoma
(Lisianthus) flower petals. (A)
GFP detection in
Osteospermum ligules
flower petals. (A)
GFP = Green Fluorescent Protein
51
4
52. Block schemes of the gfp-expression constructs used in this work.
53. GFP detection in leaves and roots of transgenic
plants. (A, B) Leaf trichomes from Osteospermum
transfected with vector alone. (C, D) Leaf
trichomes from Osteospermum transfected with
mGFP5. (E, F) Roots from Osteospermum
transfected with vector alone (left) or mGFP5
(right). (A, C, E) were illuminated with white
light; (B, D, F) were illuminated with UV light.53
GFP detection in flower and leaf extracts
PAGE analysis of plant extracts
Western blot analysis
A B
C D
E F
54. (A) Phenotypes of transgenic Torenia flowers.
• (Upper) Flower color under white light.
• (Lower) Cellular fluorescence from adaxial side of petal in each line.
(B) Expression analysis of each transgenic line by RT-PCR/Southern blotting
Coexpression of the Am4CGT and AmAS1 genes was sufficient for the accumulation
of aureusidin 6-O-glucoside in transgenic flowers (Torenia hybrida).
54Japan Ono et. al (2006)
A
B
5
56. Production of picotee-type flowers in Japanese
gentian by CRES-T
A B
Wild type flower‐Solid color suppression of pigment
production generates picotee
type flower
Nakatsuka et al., 2011Japan
CRES-TChimeric repressor gene-silencing technology
56
6
57. • (CRES-T) is an efficient gene suppression system which
worked successfully in Japanese gentian.
• A chimeric repressor of the anthocyanin biosynthetic
regulator gene GtMYB3, under the control of the
Arabidopsis actin2 promoter, was introduced into blue-
flowered gentian.
• Of 12 transgenic lines, 2 exhibited a picotee flower
phenotype with a lack of pigmentation in the lower part of
the petal.
• HPLC analysis showed that the petals of these lines
contained less anthocyanin and more flavone than the
wild-type.
• The expressions of ‘late’ flavonoid biosynthetic genes,
including F3H, F35H, DFR and ANS, were strongly
suppressed in petals of these transgenic plants.
57
58. • Expression of flavonoid biosynthetic genes in transgenic
gentian plants.
• The expression levels of GtMYB3-SRDX and endogenous
flavonoid biosynthetic genes were determined by semi-
quantitative RT-PCR analysis in wild-type and GtMYB3-
SRDX expressed transgenic gentian clone nos. 7 & 11
• Schematic representation of pSMABR-AtACT2pro-
GtMYB3-SRDX.
• Bar herbicide bialaphos resistance gene as a
selectable marker.
• NOSp promoter of nopaline synthase (NOS) gene
from A. tumefaciens.
• rbcSt terminator of RuBisCO small subunit 2B gene
from Arabidopsis.
• NOSt terminator of NOS gene.
• LB left border; RB right border.
semi-quantitative RT-PCR analysis
58
59. Flavonoid analysis in the flowers of transgenic gentian plants by HPLC
A & D- wild type
B & E- transgenic
gentian clones no. 7
C & F- transgenic
gentian clones no. 11
Anthocyanins
Flavones
59
61. Nine genes encoding putative cysteine proteases
Protein content (A) and protease activity
(B) in wild-type Petunia (MD) & 35S:etr1-
1 transgenic (etr-44568) petunia corollas
during flower development.
PhCP2 to PhCP10
61 Jones et al, 2005USA
• In this study, 35S:etr1-1 transgenic
petunias have been used to see
how ethylene regulates flower
senescence.
• To compare the senescence
programmes in ethylene-sensitive
(MD) and ethylene-insensitive (etr-
44568) flowers, a comparative
analysis was conducted of age-
related changes in total protein,
protease activity, and the expression
of nine cysteine protease genes in
the petals of MD and etr-44568
petunias.
62. ETR1-1 - Makes plants ethylene insensitive
Long Lasting Flowers
Etr1-1
62
Control Etr1-1Control Etr1-1
63. Generation of phenylpropanoid pathway-derived
volatiles in transgenic plants
• Rose alcohol acetyltransferase produces phenylethyl acetate and
benzyl acetate in petunia flowers.
• Esters are important volatiles contributing to the aroma of numerous
flowers and fruits.
• Acetate esters such as geranyl acetate, phenylethyl acetate and
benzyl acetate are generated as a result of the action of alcohol
acetyltransferases (AATs).
• To study the function of rose alcohol acetyltransferase (RhAAT), they
generated transgenic petunia plants expressing the rose gene under
the control of a CaMV-35S promoter.
• Phenylethyl alcohol and benzyl alcohol produce the corresponding
acetate esters, not generated by control flowers.
• The level of benzyl acetate is three times more than phenylethyl
acetate in different transgenic lines of petunia.
USA Pichersky et al.,200663
8
64. Molecular analyses of
transgenic petunia plants
expressing RhAAT
RNA blot
GC–MS analysis of volatile compounds
benzyl acetate
phenylethyl acetate
64
USA Pichersky et al.,2006
65. Transgenic rose lines harboring an antimicrobial
protein gene, Ace-AMP1, demonstrate enhanced
resistance to powdery mildew
• An antimicrobial protein gene, Ace-AMP1,
was introduced into Rosa hybrida via
Agrobacterium-mediated transformation.
• PCR analysis for both Ace-AMP1 and
neomycin phosphotransferase (npt II)
genes, showed that 62% of these plants
were positive for both transgenes.
• These lines were further confirmed for
stable integration of Ace-AMP1 and npt II
genes by Southern blotting.
• Transcription of the Ace-AMP1 transgene
in various transgenic rose lines was
determined using Northern blotting.
USA Li et al., (2003)65
Fig. a) Powdery mildew development
on leaflets & b) on whole leaves
Control
Control
Transgenic
Transgenic
9
66. 66
PCR analysis of transgenic plants of
Rosa hybrida cv. Carefree Beauty
Tnos Ace-AMP1 Penh-35S Pnos Npt II Tg7
EcoR 1
The binary vector pFAJ3033. The Ace-AMP1 gene is
driven by CaMV 35S with a duplicate enhancer
region, and is terminated by (NOS).
Southern blot hybridization
analysis of transgenic and control
rose plants. A) Probed with a 0.35-
kb fragment of the Ace-AMP1gene.
B) Probed with a 0.7-kb fragment
of npt II gene
Northern blot analysis whereby a 0.35-kb fragment
of the Ace-AMP1 gene was used as a probe
A)
B)
67. Conclusions
• Genetic engineering overcomes almost all the limitations
of traditional breeding approaches.
• Recent developments in plant molecular biology provide
opportunities to use techniques of genetic engineering
for improvement of flower crops for modify flower colour,
improve vase life, floral morphology, scent and disease
resistance.
• Spectral difference in flower colour is mainly determined
by the ratio of different classes of pigments and other
factors and knowledge of flower coloration at the
biochemical and molecular level has made it possible to
develop novel color.
67
68. • Recently genetic engineering has demonstrated the best
examples such as ‘Moon’ series of transgenic carnations
and transgenic blue rose marketed in North America,
Australia and Japan.
• Ethylene sensitivity regulates the timing of flower
senescence and incorporation of etr1-1 gene delayed
senescence in petunia flowers.
• Recent advances in the isolation of scent biosynthetic
genes have provided the basis and created the
opportunity for the biotechnological manipulation of floral
scent.
• Ace-AMP1 is a good candidate gene for genetic
improvement of disease resistance in roses. 68
Conclusions
69. Future prospects and new avenues
69
• New genes should be isolated that will have utility in
floriculture, and new transformation methods for flower
crops should be further optimized.
• A genetic map of rose, which is commercially the most
valuable cut flower, has now been developed.
Identification of quantitative trait locus (QTL) from this
map, in conjunction with genetic modification, will assist
breeders to improve productivity, disease and insect
resistance.
• Information on expression of regulatory genes encoding
transcription factors should be generated which have
effects on flower colour, fragrance and disease resistance.
• The manipulation of colour in the yellow and orange range
will become increasingly feasible as more is learnt about
the carotenoid biosynthesis pathway.
• More research efforts are needed to modify flower shape
Traditionally, Breeding of ornamental plants has been based mainly on continuous crossing & selection, Several decades have combined for commercially important characteristics into an elite genotypes.
They show antibacterial, antifungal, antitumor and anti-inflammatory properties.
A higher vacuolar pH (around 4.0 rather than3.0) is necessary for the blue flower colours
Fe3+, one Mg2+ and two Ca2+ ions
Anthocyanins are a red colour under low pH environments and a blue colour under neutral or alkaline pH environments
Vacuolar pH is generally regulated by vacuolar ATPase and pyrophosphase in plant cells.
Anthocyanins can be used as pH indicators because their color changes with pH. Anthocyanins are pink in acidic solutions (pH < 7), purple in neutral solutions (pH ~ 7), greenish yellow in alkaline solutions (pH > 7), and colourless in very alkaline solutions where the pigment is completely reduced.
The development of the blue carnation was not the primary goal of the research team; no, They wanted to make a blue rose but, used
stopping the production of red pigment, allowing the production of blue pigment. They were able to stop the production of the red pigment by silencing dihydroflavonol reductase (DFR), the red producing gene.
a delphindin gene from a pansy was inserted to allow blue pigment production. Finally, another DFR gene, this time from an iris, was
inserted, to actually produce the blue pigment.
Metabolic engineering is a form of genetic engineering aimed at changing the way living things metabolize, or rearrange the nutrients they take in into different chemicals and thus make useful fragrances.
Petunia flower colours are mainly due to flavonoids. Flower colour modified by the suppression of flavonoid biosynthetic genes
They showed various phenotypes.
The percentage of delphinidin in the petals of individual transgenic plants was plotted against the flower colour represented by the hue value in degrees (hue angle).
For each plant, 5 mg total RNA was subjected to RNA gel blot analysis.
CaMV35S: Cauliflower Mosaic Virus 35S promoter. E35S: enhanced CaMV35S promoter. Nos-ter: nopaline synthase terminator. Signal peptide: signal sequence of the basic chitinase of A. thaliana. Hsp70
intron: maize heat-shock protein 70 intron. ST-LS1 intron: intron of the potato ST-LS1 gene. GmHsp 17.9 leader: soybean heat-heat-shock protein 17.9 gene leader sequence. 2xE35S-pro: CaMV35S promoter with a duplicated transcriptional enhancer. pA 35S: CaMV35S poly adenylation signal. The translation start (ATG) and stop codons (TAA and TGA), the chromophore aminoacid mutations (V163A, I167T, S175G, S65T and S65C), and the HDEL endoplasmic reticulum retention signal are indicated.
Phenotypic and expression analyses of transgenic Torenia Flowers