Genetic inheritance of flower doubleness and fragrance

1. Genetic Inheritance of Flower
Doubleness And Fragrance
Submitted by,
Shreedhar Beese
Ph.D Scholar (ICAR-SRF)
UHF Solan
(Floriculture and Landscape
Architecture )

2. Introduction
Biotechnology In Floriculture
Genetic improvement of flower colour
Making deliberate crosses between two parents
Mutation
Polyploidy
Genetic Engineering of flower doubleness and Fragrance
 RNA interference technology
 Micro RNA
 Mads Box
Case studies
Limitations of plant gene transformation
Future Prospects
Conclusion
CONTENTS

3. Introduction
 Floriculture is considered to include the cut flowers, potted
plants, and ornamental bedding plants and garden plant
industries.
 Commercial floriculture is becoming important from the export
angle.
 commercial floriculture has higher potential per unit area than
most of the field crops.
 Government of India has identified floriculture as a sunrise
industry and accorded it 100% export oriented status.
 Indian floriculture industry has been shifting from traditional
flowers to cut flowers for export purposes

4.  About 255 thousand hectares area is under cultivation, and
the production of flowers are estimated to be 17.54 million
tonnes loose flowers and 543 million tonnes cut flowers.
 The country has exported 22,947.23 MT of floriculture
products to the world for the worth of Rs. 460.75 crores in
2014-15.
 The main areas of production and consumption of
floricultural products are in the United States and Europe,
 The highest consumption per head is in the Netherlands,
followed by Germany, Austria, and France.

5. Biotechnology in floriculture
oThe global flower industry thrives on novelty
oEngineered traits are valuable to either the consumer or the
producer
oFlower fragrance is one of the most important characteristics in
ornamental plants
oDevelopment of flower fragrance in ornamental plants is a major
breeding target
oPhenotype with unique forms of flower or, the double flower has
higher ornamental value than the single one
oKey transcriptional factors for the identification of floral organs
have been clarified by analysing model plants (Arabidopsis)

6. Why we need modification in fragrance
and doubleness?
• Modification in flower fragrance of a variety with desirable
agronomic or consumer characteristics
⚫Ex:Apetunia non-scented from preferable scented flowering
variety
• Aflower fragrance not occurring naturally in a particular crop
•Change in trend for Fragrance and doubleness flowers
•High price for flowers having fragrance

7. ABCDE MODEL OF FLOWER DEVELOPMENT
 This model developed on the basis of Arabidopsis
thaliana mutants. Most of the genes of ABCDE
model are MADS-box genes.
 Class A genes (APETALA1) controls sepal
development & together with class B genes (e.g.
PISTILLATA, and APETALA3), regulates the
formation of petals.
 Class B genes, together with class C genes (e.g.,
AGAMOUS), mediates stamen development.
 Class C genes determines the formation of carpel.
 The class D genes (e.g., SEEDSTICK, and
SHATTERPROOF) specify the identity of the
ovule
 Class E genes (e.g., SEPALLATA), expressed in
the entire floral meristem, & are necessary

9. Genetic Improvement Of Flower
Fragrance And Doubleness
⚫ Genetic Improvement: involves changing the plant’s genetic makeup
 Making deliberate crosses between two parents
 Conventional Hybridization
 Inter-specific Hybridization
 Mutation
 Polyploidy
o Genetic Modification
 RNAi or Gene silencing
 Chimeric REpressor gene-Silencing Technology (CRES-T)
 Micro RNA

10. HYBRIDIZATION
o Single, semi-double & double type of flower are genetically
controlled
o Based on gene governing, doubleness can be transferred into
new cultivar by hybridizing with suitable parent

12. Mutation Breeding
 The morphology of flowers &
inflorescences can be affected by
mutation
 Mutation induction led to changes in
flower size, petal shape, petal
numbers
 In Compositae, an increase in whorls
of ligulate florets as well as a
conversion from ligulate into tabular
florets was described

13. Carnation varieties co-developed by Kirin Agribio and
the JAEA using ion beams. The flower on the upper-
left corner is the parent(var. .Vital.) and the others are
mutants. Tanaka et al., 2010

14. Polyploidy
o Polyploidy breeding : Effective method to double the chromosome
number
o Genetic variations created can be further used in breeding
o Main consequences of induced polyploidy are increase in size and
shape of plants/leaves/ branches, flower parts, fruits & seeds
(Chopra, 2008)

15. Fig. Field performance of diploid and
tetraploid Gerbera jamesonii Bolus cv.
Sciella
a.Variation in plant characteristics
between diploid (2x) and tetraploid
(4x);
b. variation in stalk length between 2x &
4x;
c. variation in flower dia between 2x &
4x
Gantait et al., 2011

16. GENETIC MODIFICATION OF
ORNAMENTAL PLANTS
It offer new opportunities for breeders of ornamentalplants
Development of new ornamental varieties through gene transfer is
possible by this technique
Genetic engineering can introduce traits not be generated by conventional
breeding
Major traits amenable to manipulation by genetic modification include
flower color, fragrance, abiotic stress resistance, disease resistance, pest
resistance, manipulation of the form and architecture of plant and/or
flowers modification of flowering time, and post harvest lifeetc.
Ex: Chrysanthemum, Torenia: Fringed petal
Cyclamen, Petunia: Double flower

17. 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 DNAtechnology to alter plant characteristics.
Geneticmodificationcanbeusedtotransfernewspecifictraitsintotheplant

18. Gene transfer methods
Indirect Direct
Agrobacterium mediated
gene transfer
Most widely used
More economical
More efficient
Transformation success is 80-85%
 Particle bombardment or
micro projectile
 Direct DNA delivery by
Microinjection or PEG
mediated uptake
 Ultrasonication
 Electroporation
 Electroporotic uptake
Chandler and Brugliera, 2011

19. Gene transformation

20. RNA Interference Technology
 RNA interference (RNAi) is a
naturally occurring mechanism
that leads to the “silencing” of
genes
 In consequence, the respective
protein is not synthesized
 This technique can be used for
loss-of-function studies where a
gene is specifically silenced and
character is not expressed

21. Micro RNA
 A microRNA (miRNA) is a small non-coding RNA molecule (about 22nucleotides)
found in Eukaryotes, which functions in RNA silencing and posttranscriptional
regulation of gene expression
 miRNAs are involved in almost all biological and metabolic processes (Khraiweshet
al., 2012)
 miR156: Plant architecture (Jiao et al. 2010). miR319: Leaf & Petal
morphogenesis in Snapdragon (Carle et al., 2007)

22. MADS-BOX
 The MADS box is a conserved sequence motif found in genes which comprise the
MADS-box gene family
 The MADS box encodes the DNA-binding MADS domain
 The length of the MADS-box are in the range of 168 to 180 basepairs
 Origin:
 MCM1 from the budding yeast, Saccharomyces cerevisiae,
 AGAMOUS from the thale cress Arabidopsis thaliana,
 DEFICIENS from the snapdragon Antirrhinum majus
 SRF from the human Homo sapiens
In plants, MADS-box genes are involved in controlling all major aspects of
development, including male & female gametophyte development, embryo and
seed development, as well as root, flower and fruit development, floral organ
identity and flowering time determination

23. CASE STUDIES

24. Objectives:
1. Study of the effects of the gene silencing of C-class MADS-box genes by using a
VIGS system on flower phenotypes in petunia cultivars.
2. Comparison between Large petaloid stamens induced by silencing both pMADS3
and FBP6 with small petaloid stamens induced by silencing only pMADS3.

25. INTRODUCTION
 Double flower formation: Mainly due to conversion of stamen and
carpel into petal and new inflorescence
 Double flowers enhances the commercial value of Petunia hybrida. As
ornamental plants, double flowers with large petaloid stamens and/or
new flowers at inner whorls are desired
 C-class genes along with B-class genes, specify stamen identity in whorl 3. A/C
to ABC model of floral organ identity (Coen and Meyerowitz,1991)
 Suppressing C-class genes in whorl 3 results in the conversion of stamen into
petal. C-class genes also specify carpel identity in whorl 4 and control floral
meristem determinacy, their suppression induces the indeterminate development
of flowers in whorl 4
 C-class genes belong to AG-clade of the large MADS-box gene family

26. Materials And Methods
Plant materials:
 VIGS treatments of each of the C-class MADS-box genes, pMADS3 and FBP6, and of
pMADS3 & FBP6 conducted in four petunia cultivars, ‘Cutie Blue’, ‘Fantasy Blue’, ‘Picobella
Blue’,and ‘Mambo Purple’
Plasmid construction:
 The tobacco rattle virus (TRV)-based VIGS system (suppression of the anthocyanin pathway
via chalcone synthase silencing as reporter as it produced white flower)
Vector: pTRV1 and pTRV2 VIGS
 PhCHS was amplified and cloned into the EcoR1 site of pTRV2 vector
 The non-conserved regions of petunia C-class genes, pMADS3 and FBP6, were amplified
using the primers and cloned into the SmaI site of pTRV2 PhCHS vector individually to
generate constructs for silencing pMADS3 and FBP6 separately and fused to generate a
construct for silencing pMADS3 and FBP6 simultaneously

27. Agro-inoculation of TRV vectors:
 Virus infection was carried out by
means of the Agrobacterium-
mediated infection of petunias
 Young leaves of 3-week old petunia
plants were inoculated
Quantitative RT-PCR of C- and A-class
MADS-box genes:
Quantitative RT-PCR (qRT-PCR) of C- and
A-class MADS-box genes in petals and
stamens of VIGS-untreated control flowers
and petaloid stamens of VIGS-induced
flowers was performed.

28. Results And Discussion
• In ‘Picobella Blue’ and ‘Mambo Purple’: No white flower was noted (Unknown genetic
background, Chen et al., 2004)
• In ‘CutieBlue’and ‘Fantasy Blue’: Completely white double flowers were
observed, indicating the strong and complete silencing
• In flowers inoculated with either pMADS3-VIGS orFBP6-VIGS,
morphologically significant but small conversions in whorls 3 & 4 were observed
• In flowers of pMADS3-VIGS inoculated petunias, anthers converted into small petaloid
tissues but filaments retained their original struc (Fig. 1c & d)
• In flowers of FBP6-VIGS inoculated petunias, the stamens were almost
unaffected
• In petunias inoculated with pMADS3/FBP6-VIGS, prominent double flowers with
highly ornamental appearance formed. Complete loss of stamen identity was observed.
Both anthers and filaments were completely converted into petaloid tissues

29. Fig. 1. Morphological changes in
flowers of P.hybrida cv ‘Cutie Blue’
inoculated with pTRV2-
PhCHS/pMADS3 (pMADS3-VIGS)
and pTRV2-PhCHS/pMADS3/FBP6
(pMADS3/FBP6-VIGS).
(a) VIGS-untreated control flower;
(b) Stamens and a carpel of non-
VIGS
flower;
(c) pMADS3-VIGS flower (white
and blue mixed color);
(d) Petaloid stamens and a carpelof
pMADS3-VIGS flower;
(e) pMADS3/FBP6-VIGS flower
(white);
(f) Petaloid stamens and a carpel of
pMADS3/FBP6-VIGS flower
(white).

30. Fig. 2. 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) stamens and carpels or converted
new flowers of
pMADS3/FBP6-VIGS flowers.

31. Flowers inoculated with pMADS3/FBP6-VIGS in whorl 4, carpels converted into new flower
(Cultivar-dependent)
In 50% of the double flowers of ‘Mambo Purple’, a 2nd new flower arose instead of a carpel. This
process was repeated, generating 3rd new flowers. It exhibited avoluminous and decorative
appearance with a high commercial value.
Fig. 3. New flower formation in whorl 4 and from axil
of whorl 3 in a double flower of P. hybrida cv ‘Mambo
Purple’ inoculated with (pMADS3/FBP6-VIGS).
An opened double flower with asecond new flower in
whorl 4 An opened second new flower;
Fused corolla (left), a carpel (center), and petaloid
stamens (right) of the second flower;
An ectopic new flower emerging
from the axil of whorl 3;
An unconverted stamen (left) and petal-like tissues of
the ectopic new flower.

32. The surface areas of petaloid stamens in pMADS3/FBP6-VIGS plants were
more than 10 times as large as those in pMADS3- VIGS plants

33. Double flowers can be induced by virus-induced gene silencing (VIGS) of
two C-class MADS-box genes, pMADS3 and FBP6
Large petaloid stamens induced by pMADS3/FBP6-VIGS were compared
with small petaloid stamens induced by pMADS3-VIGS
New flower formation in the inner whorl of flowers silenced in both
pMADS3 and FBP6 gene is cultivar-dependent
They are valuable for future breeding of petunia cultivars bearing decorative
double flowers with large petaloid stamens and inner new secondary flowers

34. A miR172 target-deficientAP2-like gene correlates with the
double flower phenotype in roses
Case Study -2
 One of the well-known floral abnormalities in flowering plants is the double-flower phenotype, which
corresponds to flowers that develop extra petals, sometimes even containing entire flowers within
flowers. Because of their highly priced ornamental value, spontaneous double-flower variants have
been found and selected for in a wide range of ornamental species.
 double flower formation in roses was associated with a restriction of AGAMOUS expression domain
toward the centre of the meristem, leading to extra petals.
 An APETALA2-like gene (RcAP2L), a member of the Target Of EAT-type (TOE-type) subfamily, lies
within this interval.
 In the double flower rose, two alleles of RcAP2L are present, one of which harbours a transposable
element inserted into intron 8.

 This insertion leads to the creation of a miR172 resistant RcAP2L variant.
www.nature.com/scientificreports/

35. Figure 3. The transposable element
insertion is only observed in the
analysed double flower roses. PCR
to detect the TE were performed on
genomic DNA from different rose
cultivars exhibiting simple (b) or
double flowers deriving from R.
chinensis (a). The lower band (419
bp) corresponds to the combination
of primers that amplifies the wild
type allele, while the two higher ones
(770bp and 754 bp) correspond to
amplification of the left and right
borders of the transposable element
insertion, respectively.
www.nature.com/scientificreports/

36. Model showing how a miR172-resistant euAP2 could lead to double flower formation. (a) In wild- type flowers, euAP2 are expressed
in the first and second whorls where they can inhibit RcAG expression likely by recruiting cofactors and histone modifiers, such as
TOPLESS and HDA19. Sepals and petals are consequently formed. In the 3rd whorl, miR172 is expressed and inhibits euAP2 proteins
production, releasing the inhibition of RcAG. RcAG will then determine stamens and carpels identity and formation. (b) In double
flowers, the truncated version RcAP2L 172 mRNA (following TE insertion) is insensitive to miR172 inhibition. RcAP2L 172
expression is maintained in the meristem and down-regulates RcAG expression.

37. Genotype Flower multiplicity
Ploidy level
WT 5′ of TE 3′ of TE
R. chinensis ‘Old Blush’ Double flower 2x 5.7 7.2 7.4
R. chinensis homozygous genome NA 2x 0 22.2 16.7
R. odorata ‘Hume’s Blush’ Double flower 2x 8.4 9.2 6.9
R. x hybrida ‘La France’ Double flower 3x 5.1 5.1 8.0
R. chinensis ‘Sanguinea’ Simple flower 2x 22.5 0 0
R. chinensis ‘Spontanea’ Simple flower 2x 13.2 0 0
R. wichurana Simple flower 2x 8.6 0 0
Identification of RcAP2Lalleles present in 7 re-sequenced genotypes. Number of 100 bp genomic reads
overlapping intra-gene (denoting the presence of a wild-type RcAP2L allele) or gene-TE (mutated allele,
RcAP2L) junctions, for genotypes with double or simple flowers. The read counts were normalized according to the
read library size, and expressed as reads per 100 million reads. Ploidy level
Raymond et al.,

38. Analysed the presence of this variant in a set of simple and double
flower roses demonstrate a correlation between the presence of this allele and
the double flower phenotype. These data suggest a role of this miR172 resistant
RcAP2L variant in regulating RcAGAMOUS expression and double flower
formation in Rosa sp.
Conclusion
www.nature.com/scientificreports/

39. Tinkering with the C-Function:A Molecular Frame for the Selection of
Double Flowers in Cultivated Roses
Case Study -3
Introduction : In ornamental plants, flower traits such as the floral architecture, petal
color and recurrent flowering are key characters that have been subjected to artificial
selection pressure during the early domesti- cation and the subsequent breeding process.
Flower forms with increased number of petals (termed double flowers) were retained for
their showy aspect in many domesticated plant families. In Rosaceae, for instance,
spontaneous double flower forms were kept and propagated for garden ornament (Prunus,
Rosa, Potentilla…). Rose species were domesticated several times independently. The two
major areas of rose domestication in the Antiquity were China and the peri-mediterranean
area (encompassing part of Europe and Middle East), where R. chinensis Jacq. and R. gallica
L. were bred and contributed predominantly to the subsequent selection process. In both
cases semi-double (8 to 40 petals) and double flowers.
www.plosone.org

40. Simplified genealogy of roses
Cultivated roses originate from two main
regions of domestication,
i.e. the peri-mediterranean areas
(Europe/Middle-East) and China. Double
flowers were selected independently in the
European and Chinese lineages.
‘Cardinal de Richelieu’ and ‘Old Blush’
represent examples of double and semi-
double flower varieties in the R. gallica and
R. chinensis lineages.
These two gene pools were kept separated
until the early nineteenth century, when
they were crossed to obtain triploid hybrids
and tetraploid modern varieties.
www.plosone.org

41. Figure 3. Floral organ numbers in
‘‘Malmaison’’ and ‘‘St Anne’s’’.
(A) Longitudinal sections of flower in
‘‘Malmaison’’ (left) and in ‘‘St Anne’s’’ (right).
(B) Floral organs number in ‘‘Malmaison’’ (dark
grey) and in ‘‘St Anne’s’’(light grey).
Histograms represent the means obtained from 5
flowers from each hybrid. Error bars represent
the standard deviation. The two rose varieties
differ in two floral characters: organ identity
reversions from petals in ‘‘Malmaison’’ to
stamens in ‘‘St Anne’s’’ and an overall decrease
in total organ number. Chimeras: staminoid
petals (see Figure S1).
(C) Bivariate plot of petal and stamen number
showing anti-correlation in ‘‘Malmaison’’
flowers, thus the lability of petal/stamen
boundary in this genotype. Each square
represents one flower. Correlation and
determination coefficients are R=20.84; R2
=
0.71.

42. Longitudinal sections of floral meristems and flowers during floral organogenesis. (A–J) Sections (stained with toluidine blue) of ‘‘Malmaison’’ (A–D,I) and ‘‘St
Anne’s’’ (E–H, J) were observed, from the floral meristem stage (stage 1;A, E)until carpel formation (stage 4, I,J). Scale bar equals 150 mm for A to H and 1 mm for Iand
J. (K) Analysis of floral organogenesis in ‘‘Malmaison’’ (top) and ‘‘St Anne’s’’ (bottom). Sepals, petals, stamens and carpels are labeled in yellow, green, blue and red
colors, respectively. The different whorls composition is displayed as follows: whorl 1 comprises 5 sepals; whorl 2 is composed of the first 10 petals; whorl 3 is
composed of stamens in ‘‘St Anne’s’’ and petals plus stamens in ‘‘Malmaison’’; whorl 4 is composed of carpels. Numbers 1 to 5 at the bottom define the flower
development stages. Note that ‘‘Malmaison’’ has an enlarged floral receptacle starting from stage 4 (I).

44. Model for selection of double roses.
In wild-type roses (a):
the petal/stamen boundary is very
stable, as all wild species have 5 petals.
In cultivated roses, the petals/stamens
boundary is labile within the flowers.
Breeders have tinkered with this
instability of petals/ stamens boundary
by acting on expression domain of the
rose ortholog of AGAMOUS, all along
breeding history to select either for semi-
double flowers (b) or double flowers (c).
www.plosone.org

46. Genetic Engineering For
Floral Scent
may enhancethevalueofcutflowerstoconsumers…
Fragrance numerous volatile
aromatic organic substances
present in the flower.
Such as,
hydrocarbons,
alcohols,
aldehydes,
ketones, esters,
ethers
Manipulation of fragrance inflowers
chemicals contributing to the fragrance of
roses, their pathways of synthesis and
enzymes controlling these pathways to be
identified.

47. Floral Scent Modification
• Secondarymetabolites
Volatile, low-molecular-weight, givethe flowers their unique,characteristic
Fragrances
• Typesof scent compounds:
Class Precursor Types Examples
Terpenoid
Phenylpropanoids
(benzoids )
Fatty acid
derivative
s

48. Genesresponsibleforscentproduction
Flower crop Genes responsible floral volatiles Reference
Clarkia breweri
(S)-linalool synthase (LIS)gene Dudarevaet al, 1996
Isoeugenol-O-methyltransferase (IEMT) Wanget al, 1997
Benzylalcohol acetyl-transferase(BEAT) Dudarevaet al, 1998
Salicylicacidcarboxyl methyl transferase
(SAMT)
Rosset al,1999
Benzoicacidcarboxyl methyl transferase
(BAMT)
Picherskyand Dudareva,
2000
Benzylalcohol benzyltransferase (BEBT) Dudarevaet al, 2000
Petunia hybrida
P. axillaris
Benzoicacid/salicylic acidand carbonyl
methyl transferase(BSMT)
Negreet al, 2003
Benzylalcohol /phenyl ethanolbenzyl
transferase (BPBT)
Boatright et al,2004

49. Rosa hybrida
Germacrene D synthase Gueterman et al, 2002
Geraniol/citronellol acetyl transferase Shalit et al, 2003
Benzyl alcohol /phenyl ethanol benzyl
transferase (BPBT)
Boatright et al, 2004
Orcinol-O-methyl transferase (OOMT)
R. chinensis Phloroglucinol-O-methyl transferase
(POMT)
Lavid et al, 2002
Antirrhinum majus
(Snapdragon)
Mycene synthase
Ocimene synthase
Dudareva et al, 2000
Stephanotis floribunda
(Madagascar Jasmine)
Salicylic acid carboxyl methyl
transferase (SAMT)
Pott et al, 2002
Arabidopsis thaliana (S)-linalool synthase (LIS)
Caryophyllene methyl transferase
(BSMT)
Chen et al, 2003
Vanda Mimi Palmer Linalool synthase (LIS), acetyl-CoA
acetyltransferase (ACA), 1-
deoxy-D-xylulose 5-phosphate synthase
(DXPS), 3-hydroxy-3-methylglutaryl-
coenzyme A reductase(HMGR)

50. ODORANT1 regulatesfragrance in petuniaflowers cv.Mitchell
• Petunia hybrida : volatile benzenoids ODORANT1(ODO1)
•
• Flowers fragrant in the evening and atnight
• Transcript levels of ODO1 before the onset of volatileemission
decreased when volatile emission declined
ODO1 transgenic P. hybrida Mitchell benzenoid levels synthesis of
precursors from shikimatepathway

51. Volatile benzenoids emission by Mitchell (M), RNAi lines (1,3,12,35)
Verdonk et al, 2005

52. PAP1enhances phenylpropanoidandterpenoidproductionin Rosahybrida
cv.PariserCharme
• ArabidopsisPRODUCTIONOFANTHOCY
ANINPIGMENT1(P
AP1) Rose
• PAP1-transgenicrose lines phenylpropanoid (color andscent) when
compared with control flowers(GUS)
• PAP1- lines6.5 times terpenoid (scent)
Development of plant from somatic embryo
GUSTransgeniccontrol
GUSTransgeniccontrol following
X-Gluc staining
PAP1Transgenicline
i, ii,iii
iv, v,vi
iv, v,vi

53. Thelevels of emission ((e), µgper flower per 24h) and internal pools ((p), µgper flower) of volatile
compounds produced by flowers of PAP1-transgeniclines 6, 11and 12when comparedwith control
Zviet al, 2012

54. Clarkia
breweri
benzyl alcohol acetyltransferase
(BEAT)
Eustoma
grandiflorum
No benzyl acetate
No fragrance
BEA
Tcatalyzing the synthesis benzyl acetate which constitutes up to 40%of C.breweri’s
total scent output (Dudareva et al,
1998)
benzyl alcohol acetyltransferase
(BEAT)
Alcohol
substrate 5-7 times higher levels of
benzyl acetate
Fragrant
Aroma enhancement in transgenic Lisianthus using the Clarkia Beat gene

55. Control and transformedadult
flowering plants
Levelof benzyl acetate in control and
transgenic lines after feeding with BAor
water- (A)Leaf (B)Flower
Aranovich et al, 2007

56. Linalool synthase
(LIS)
Linalool glucoside
Luckeret al, 2001
Linalool
Lavyet al, 2002
Strawberryalcohol
acyltransferase
(SAAT)
Isoamyl alcohol
Acetyl esteracc.
Volatile unaltered
Beekwilder et al,
2004
Alcohol
acetyltransferase
Acetate ester acc.
Alcohol
substrate
Guterman et al,
2006

57. Commercialization Hurdles- For GE Ornamentals
Crops
• Ornamentalshavemuch smaller market value than foodcrops
• Highcost of analysis,risk assessmentand regulatoryapproval
- Supercarnations, color-modified Torenia
• Regulatoryapproval for field testingtakesmonths or years
• Molecular characterization require PCR-basedidentificationtest for
which fee of 30,000EURO
Chandler, 2013

58. www.burpee.com
Marigold- FrenchVanillaHybrid
Burpee SeedCompany., USA-creamy-
white fully doubleflowers

59. Cv. Y
ellow
Baby
Sensation Cinderella Double
Pink
Super
Gold
Flower Single Single Single Double Single
Flower
colour
Yellow Pink Lavender
pink
Pink Dark
yellow
Bulb
size
10cm 10cm 10cm 12cm 12cm
Stem
length
35cm 45cm 35cm 65cm 65cm
Ludwig & Co.,Holland, 2013
www.floraculture.au
Heavilyscentedandboldlycoloured tuberose

60. 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.
• 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.

61. FUTURE PROSPECTS AND NEW
AVENUES
• 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 doubleness and fragrance.
• More research efforts are needed to modify flower fragrance
and doubleness

62.  GEisbreeding tools that future generations canusetotackle
environmental challenges
 Nogeneticbarriers
 Creation of geneticvariation
 Novelty through geneticengineering
 Speedof improvement
 Altered plant byproduct, form and scent
 Knowledge at the biochemical and molecular level has made it
possible to develop novel colour which are otherwise absent in
nature.
 Transgenic floricultural crops, only few crops.indicating
development of commercial crops by GE is still very
challenging.
Conclusion

63. THANK YOU…