1) Safflower is an annual herbaceous plant cultivated for edible oil, herbal tea, and textile dye. It faces challenges like low seed yield and lack of disease resistance. 2) Genetic improvement strategies include developing ideotypes for high seed yield through heterosis, interspecific hybridization, and increasing oil content. Marker-assisted breeding is being used to select for high oleic acid and disease resistance traits. 3) Studies have identified microRNAs related to high oleic acid content and developed disease-resistant plants using tissue culture selection. Genetic transformation protocols allow incorporation of useful genes.

1. Presented by:
Yengkhom Linthoingambi Devi

2. • Scientific name: Carthamus tinctorius
• Common name: kardai, kusum
• Chromosome number: 2n=24
• Family: Compositae
• Morphology: Highly branched,
herbaceous, thristle-like annual plant.
plants with 30-150 cm tall with globular
flower head having yellow, orange and red flowers.
Uses: for edible oil
for herbal tea
for dyeing purpose in textiles

3. CURRENT CHALLENGES
• Lack of existent appropriate ideotypes for producing
seed and enhancing oil seed safflower.
• Manual harvesting problems due to spiny genotypes.
• The narrow genetic diversity for genotypes with high
acclimatization to different growing condition and
extreme temperatures.
• Low harvest index.
• Deficiencies in breeding lines containing tolerance to
different diseases and pests.

4. Genetic improvement strategies
1.Seed yield improvement
a) Ideotype change:
Induction of suitable change in different morphological, phenological, and agronomic
triats could significantly increase the seed yield.
b) Heterosis:
Production of hybrid cultivars with superior heterosis would be facilitated by using
cytoplasmic male sterility and genetic male sterility.
c) Interspecific hybridization:
Species with same chromosome number could produce fully fertile and
Hybrids in F1 andd F2 generations could transfer suitable traits to the cultivated
safflower.

5. Oil and meal improvement
• Increasing oil content has lead to
the release of cultivars like NARI-6,
NARI-NH-1, NARI-57.
• Different methods like gene
transfer, mutation, crossing with
low hull content and high oil
content genotypes.

7. Genetic analysis
• After 1990s importance of using markers for genetic diversity study is
being realized and started using it.
• Assessment of genetic diversity in safflower was mainly focused on
estimating genetic variation by combining molecular polymorphism
and phenotypic variation (Johnson et al., 2007)

8. Marker-assisted breeding
• Recently, sequence characterized amplicon region (SCARs) markers based on RAPD
polymorphism were developed for the closely linked recessive monogenic genes Li
(controlling very high linoleic acid content) and Ms (controlling nuclear male
sterility) (Hamdan et al., 2008).

9. Findings:
• Bulked segregant analysis involving a population of 162 individuals from a cross
between CLI (NMS) and CR 142 (high linoleic acid) led to construction of a linkage
map with five RAPD-SCAR markers.
• The SCAR markers flanked both loci at 15.7 cM from the Li locus and 3.7 cM from
the Ms locus. Recessive genetic male steriles are propagated through
heterozygotes (Msms) which can be identified only by progeny testing.
• Mapping of nuclear male sterility gene allows early identification of lines
carrying the male sterile allele precluding the need for progeny testing.
• Markers linked to high linoleic acid content will facilitate marker assisted
selection programme aimed at introgression of Li alleles into desirable agronomic
background.
• This study serves as a prelude for development of markers linked to
agronomically desirable traits in safflower for accelerating the breeding
programmes.

10. Genomics
• Safflower crop suffers from lack of well-developed genetic
resources. As of now, 285 nucleotide sequences and 41,000 ESTs are
reported in safflower through subtractive genomic library and
composite data base.
• Isolation of SSR markers is laborious, time-consuming and an
expensive process. However, with the availability of ESTs for
safflower it is possible to identify genic SSRs enabling the mapping
of genes of known function.
• The vast genomic resources from other compositae members viz.,
sunflower and lettuce provide a potentially valuable source for
mining SSR markers. The transfer success of Helianthus SSRs to
safflower was 13%.

12. They studied the molecular feature of high oleic acid traits in safflower by profiling the
miRNA population in developing safflower seeds expressing the ol alleles in comparison to
the wild type high linoleic safflower using deep sequencing technology.
The oleic acid content in the classic safflower oil is normally 10–15% while a natural
mutant (ol) accumulates elevated oleic acid up to 70% in seed oil.
∼16.5% of the unique small RNAs were overlapping in these two genotypes.
Target genes with conserved as well as novel functions were predicted for the conserved
miRNAs.
13 miRNAs are identified which differentially expressed between the HO and HL
safflower genotypes.
They identified two candidate novel miRNA family identified to be likely unique to the
developing safflower seed.

13. Tissue culture
• Safflower can be regenerated through organogenic and embryogenic pathways
through direct and callus mediated methods (Reviewed in Sujatha, 2007).
• Recently, the tissue culture technique has been extended for production of plants
resistant to Alternaria carthami (Vijaya Kumar et al., 2008).
•
• In this protocol, embryogenic and organogenic calli were subjected to selection
of shoots on medium supplemented with 40% fungal culture filtrate of A.
carthami (5 x 105 conidia/ml).
• Resistance in selected plants was increased to 100, 97.6 and 84% over that of the
control in R0, R1 and R2 generations, respectively.

14. RESULTS:
1.Alternaria carthami fungal spores sprayed on the leaves of FCF-tolerant plants
showed enhanced survival rate over control plants.
2.The number of leaf spot lesions per leaf was decreased from 3.4 to 0.9 and
their lesion length was also reduced from 2.9 to 0.7 mm in organogenic derived FCF-
tolerant plants over control.
3. In somatic embryo derived FCF-tolerant plants, the number of lesions was
decreased from 3.1 to 0.4 and the lesion size was also reduced to 2.7–0.5 mm when
compared to the control.
4. The study also examined antioxidant enzyme activity in FCF-tolerant plants.

15. Genetic transformation
• In safflower, Agrobacterium-mediated transformation protocols have
been reported for both Indian and American cultivars (Orlikowska et
al., 1995; Rohini and Rao, 2000).

16. Results :
• Efficient and reproducible protocol has been developed with a
transformation efficiency of 4.8% and 3.1% for S-317 (high oleic acid
content) and WT (high linoleic acid content) genotypes respectively.
• An improved safflower transformation T-DNA vector was developed,
including a secreted GFP to allow non-destructive assessment of
transgenic shoots.
• To overcome poor in vitro root formation for the first time a grafting
method was developed for safflower in which ~50% of transgenic shoots
develop into mature plants bearing viable transgenic T1 seed

17. Results revealed that Carthamus tinctorius has significant hypoglycemic
effect at 200mg/kg and 300mg/kg doses as compared to diabetic
control group. Insulin levels were significantly increased in
Glibenclamide treated as well as Carthamus tinctorius treated groups as
compared to diabetic control.
PHARMACEUTICAL

18. CONCLUSION
• The availability of extensive ESTs resources within the
Asteraceae members will facilitate comparative mapping
studies and in development of SSR markers.
• Identification of suitable markers for identification of
heterozygous maintainer in GMS and the fertility restorer
genes in CMS system will add new dimension for hybrid
breeding programmes.
• Development of mapping population through doubled haploid
technique would be relatively easy in safflower owing to the
propensity of safflower anthers for shoot regeneration