Breeding for quality improvement in Brassica has focused on developing varieties with low erucic acid (<2%) and low glucosinolate content (<30 μmol/g) through conventional and molecular breeding approaches. Promising varieties developed in India include BPRQ-2-1-5 and BPRQ-2-2-11 with low erucic acid (<2%), low glucosinolate (15.7 and 22.1 μmol/g respectively), and high yields of over 1800 kg/ha. Marker assisted selection has been used to introgress low glucosinolate genes from B. oleracea into B. rapa, reducing 5C aliphatic

1. PROSPECTS AND BREEDING QUALITY
IMPROVEMENT OF BRASSICA
Presented
By
Mohammad Jafar Tanin
Regd. No. J-14-M-368
Division of Plant Breeding and Genetics
Sher-e-Kashmir University of Agricultural Science & Technology,
Jammu

2. WHAT IS QUALITY?
Agriculture must now focus on a new paradigm that will not only produce more food,
but deliver better quality food as well
Quality refers to the suitability or fitness of an economic plant product in relation to
its end use.
Definition - According to our needs from the viewpoint of seeds, crop growth, crop
product, post-harvest technology, consumer preferences, cooking quality, keeping
quality , transportability, etc., (Gupta, 2001)
QUALITY TRAITS
A trait that defines some aspect of produce quality is called quality trait.
Each crop has a specific & often somewhat completely different set of quality traits.

3. Quality in Brassica?
Meal qualityOil quality

4. Why
quality in
Brassica
?1. Better human nutrition
2. Better animal nutrition

5. Enhancement of Nutritional Quality
Nutritional quality of oil and cake is of prime importance due to its direct and indirect impact on
human and animal health.
Brassica oil is nutritionally superior to most other edible oils due to lowest amount
of saturated fatty acids(SFA) and good proportion of mono- and polyunsaturated
fatty acids.
Oil seed Brassica meal is an inexpensive protein-rich food for animal.

7. Oil quality
 Saturated fatty acids (SFA): palmitic acid increases the LDL in blood that has a
significant role in cholesterol deposition.
 Mono unsaturated fatty acids(MUFA): Oleic acid being thermostable provides a
longer shelf life and are preferred for cooking and deep frying . It also reduces
the cholesterol . High eruic acid content has been reported to cause impaired
myocardial conductance and increased blood cholesterol.
 Poly unsaturated fatty acids(PUFA): Linoleic and Linolenic acids two essential
fatty acids which are not present in some other oils.
Our objective is to develop the varieties with
Increased oleic acid level: to provide longer shelf life
Zero erucic acid : to reduce the level of blood
cholesterol

8. Ideal fatty acid composition for safe human health
Fatty acid Per cent of the total fatty acid
Erucic acid >2
Palmitic acid 6
Oleic acid 65
Linoleic acid 20
Linlenic acid 9
Downey (1990)

9. Desaturation
ChainElongation
Lauric acid
C 12:0
Myristic acid
C 14:0
Palmitic acid
C 16:0
Stearic acid
C 18:0
Oleic acid
C18:1
Lenoleic acid
C18:2
Linolenic acid
C18:3
ssStearoyl-ACP
desaturase
Or
∆9 desaturase
Oleoyl-ACP desaturase
Or
∆12 desaturase
Eicosenoic acid
C20:1
Erucic acid
C22:1
Fatty acid Biosynthesis pathway in rapeseed mustard

10. Fatty acid composition (%) Source
Species Palmitic Oleic Linoleic Linolenic Erucic
B. napus 2.9 16.3 14.0 9.9 44.2 Stefansson, 1983
B. juncea 3.9 14.5 13.9 13.7 48.0 Munshi et al., 1990
B. rapa 3.5 13.1 13.7 14.4 51.3 Sukhija et al., 1984
B.
campestris
( zero
erucate)
3.5 64.5 22.0 10.0 <1 Stumpf and Pollard,
1983
B. napus
(Zero
erucate)
3.8 62.7 19.9 19.9 0.2 Stefansson, 1983
Soybean 19.8 31.3 43.2 4.2 - Singh et al., 1981
Sunflower 4.2 50.0 42.4 - - Sukhija et al., 1980
Groundnut 15.0 43.6 34.9 - - Randhawa, 1984
Safflower 5.0 6.1 74.9 - - Doulatabad et al., 1982
Variability for fatty acid profile in oilseed brassicas

11. Sr.
No
Type Use
1 Zero erucic acid (<2%) Nutritionally superior
2 High linoleic acid (40-50%) Nutritionally superior
3 Oleic acid (upto 70%) Nutritionally superior
4 High petroselinic acid Polymers
5 Very low lenolenic acids (<3%) Prolonged shelf life, margarines
6 High erucic acid (40-50 %)
(>80%)
Industrial polymers, lubricants,
plastic industries
Cosmetics and pharmaceuticals.
7 High stearic acid (20-40 %) Margarines
Diversified usage of oil with modified fatty acid composition
Chauhan et al. (2002)

12. Nutritional quality improvement
Donors for zero erucic acid
B. napus : Bronowski and Duplo
B. juncea : Zem-1 and Zem-2
B.rapa : Span and Tower
Some of the low erucic/ high oleic acid strains of rapeseed- mustard registered at NBPGR, ICAR
are TERI(OE)M21, TERI(OE)R03, TERI(OE)R09
B. juncea : Several low erucic acid genotypes such as TERI(OE)M21, TERI(OE)M9901,
TERI(OE)M9902, LEB-15, LES-39, PBCM-8-2, YSRL-9-18-2, CRL-1359-19, PRQ-9701;
B.napus: low erucic acid genotypes TERI(OE)R03, TERI(OE)R05, TERI(OE)R15,
TERI(OE)R983, TERI(OE)R984; have been developed and tested under the All India
Coordinated Research Project on Rapeseed-Mustard (AICRPRM), ICAR.
Recently TERI- Unnat (INGR No. 98001)and B. juncea strain LES-39 have been released.
Developed through inter specific/inter generic
hybridization followed by pedigree method of
breeding

13. Meal quality
Brassica meal contains about 40% protein with a well balanced amino acids and is an
excellent source of proteins valued for animal nutrition (Miller et al. 1962).
Its feeding value is limited due to sulphur containing compound called glucosinolate.
The break down products adversely affect the iodine uptake by the thyroid gland
Glucosinolate
C –– R
SO3
- –– O –– N
C6H12O5 –– S
Myrosinase
H2O
R –– NCS
R --- NCS
isothiocyanate
R --- NC + R --- SH
nitrile
R -- SCN
thiocyanate
+ H2O + C6H12O6
Source: Zukalova and Vasak (2002); Kaushik and Agnihotri (1999)

14. Species/crop Varieties Glucosinolate
content (µmol/g)
Donor for zero/low
content
B. campestris TL 9001 26.79 Bronowski
Heera
B. juncea RL 1359 57.14
RSPR 03 112.68
Varuna 115.28
B. napus GSL 1 69.64
Source: Rani, 1994; AICRP(R&M), 1999, 2001
Our objective is to develop the varieties with < 30 µmol/g
defatted meal and zero erucic acid known as 00 cultivars. The
biochemical composition of presently cultivated varieties
containing high erucic acid and high glucosinolate does not
match with internationally accepted standards
Glucosinolates in oilseed brassicas

15. Fatty acid profile and glucosinolate content in low erucic acid and double low rapeseed-
mustard varieties released in India
Priyamedha et al, 2015

16. What is “Canola” or “00”?
Varieties having oil with < 2% erucic acid and glucosinolates < 30 µ moles/gm in defatted
seed meal are termed as “Canola” (00)
The first zero erucic acid strains of Brassica napus were
developed in 1961. This was followed by the development of
zero erucic acid strains of Brassica campestris.

17. Rapeseed oil modification
Modified by changing the fatty acid composition to improve the edible qualities by
following the conventional and advanced breeding techniques
The most impressive example of conventional genetic manipulation is canola-a new
oil seed crop which contains <2 % erucic acid and < 30 µmol/g glucosinolates in
defatted meal obtained through crossing mutant with intermediate to low erucic acid
content.
Recent molecular analysis showed that this canola plant contains single point
mutation encoding isoforms of the enzyme β-ketoacyl CoA synthase
This fatty acid complex mediates the formation of erucic acid from lenoleic acid
Our modern canola/rapeseed varieties
are the outcome of the alteration of just
two nucleotides in a genome that
contains over I.2 billion nucleotidesThis demonstrates The power of genetics

18. Desaturation
ChainElongation
Lauric acid
C 12:0
Myristic acid
C 14:0
Palmitic acid
C 16:0
Stearic acid
C 18:0
Oleic acid
C18:1
Lenoleic acid
C18:2
Linolenic acid
C18:3
ssStearoyl-ACP
desaturase
Or
∆9 desaturase
Oleoyl-ACP desaturase
Or
∆12 desaturase
Eicosenoic acid
C20:1
Erucic acid
C22:1
Fatty acid biosynthesis pathway in canola
“canola block”

19. The Fathers of Canola
Dr. Baldur Stefansson
In 1974, Dr. Baldur Stefansson,
Professor at university of Manitoba,
developed first double low napus
variety; Tower
Dr. Keith Downey
Dr. Keith Downey, Emeritus scientist at Agri-food Canada
showed that pathway for elongation of oleate to erucate
was controlled by only a few genes and thus can be
manipulated by classical mutation breeding

20. In addition to the low erucic acid and low
glucosinolate, yellow seed coat colour is another
desired characteristic and during recent years the
concept of 'OO' is being expanded to 'OOO' to include
this as one of the major breeding objectives
Advantages of yellow coloured seed coat
Better meal quality due to low crude fibre content.
Discoloring of oil and lecithin is avoided, and light coloured end product are
obtained even without dehulling of seeds.
Yellow seeds contains normally 1-2 per cent extra oil in the embryo because the
yellow seeds possess a thinner seed coat and higher oil content in the embryo.

21. Low erucic acid and/or low glucosinolate rapeseed-mustard germplasm registered
in India
Priyamedha et al, 2015

22. Breeding
Approach
Variety Trait
Development
(%)
Reference
Selection
Horsham
Erucic acid 0.69
Burton et al. (1999)Oleic acid 54.73
Beulah Erucic acid 0.47
Oleic acid 44.72
Backcross
Swarna x Rajat Erucic acid <2
Chauhan et al. (2004
BJ 1058 x Rajat Glucosinolate <30 μ mole/g
Mutation
Abasin-95 Erucic acid 2.98 Shah et al. (2001)
Glucosinolate 25.0
MAS Varuna×Heera Erucic acid FAE1 gene
Bhatia and Alok et al.,
2014
Genetic
Transformation
Maplus
Erucic acid
<2
Zebarjadi et al., 2006
Some examples of breeding approaches using for quality improvement
in Brassica

23. Variety Trait Development
(%)
Reference
VH 486
High oleic acid 73
Sivaraman et al., 2004
Low erucic acid 0
Tammi High oleic acid 78 Jung et al., 2011
HO 1036 High oleic acid 71.71 Guan et al., 2012
N2-3591 High oleic acid >75 Nabloussi et al.,
PHOP-2-2
High oleic acid 70.1
Sachan et al., 2007
Low erucic acid <2
FFRP4-4
High oleic acid 85
Peng et al., 2010
Low erucic acid Very low
BPRQ-2-2-11
Low erucic acid 0.95
Supriya et al., 2014
Low Glucosinolate
22.1 μmol/g
Selected examples of some of important research on development of
high oleic acid and low golucosinolate and erucic acid

24. Variety Trait Development (%) Reference
Xiangyou High oleic acid 71 Xiao et al., 2009
(BLN1239)×(RI 25) High oleic acid 89
Stoutjesdijk et al.,
2000
Maplus Low erucic acid 1.1
Zebarjadi et al.,
2006
BPRQ-2-1-5
Low erucic acid <2
Supriya et al., 2014Low
Glucosinolate
15.7 μmol/g
Brink
Low erucic acid <2 Jonsson et al., 1977Oro
Span
(RI 16) ×(Sowball 76)
Low
Glucosinolate
20 μmol/g Hirani et al., 2013
(Rl1359) ×(NUDH-YJ-
04)
Low
Glucosinolate
69.39 μmol/g Jonsson et al., 1977

25. The present study aimed at development and evaluation of double low quality lines (seed
oil with < 2% erucic acid and glucosinolate content < 30 μmoles/gram defatted seed meal)
in Indian mustard. A total of 11 double low lines were selected from among a pool of 1200
lines from the cross (Varuna×NDUHYJ-3) in F7 generation. Amongst the selected lines, 2
lines BPRQ-2-1-5 and BPRQ-2-2-11 were found to be highly promising in terms of oil
quality and yield performance.
BPRQ-2-1-5 : (1884 kg/ha yield),( 15.7 μmol/g glucosinolate) and (1.1% erucic acid)
BPRQ-2-2-11: (1851 kg/ha yield), (22.1 μmol/g glucosinolate) and (0.95% erucic acid)

26. Schematic diagram of the breeding methodology adopted in the study

27. A resynthesized B. napus line, from a cross between a high glucosinolate content B.
rapa double haploid (DH) line, RI 16 and B. oleracea white cauliflower accession
Snowball 76, was backcrossed with B. rapa, RI 16. Marker assisted selection for non-
functional gene was performed in each backcross generation. Reductoin in 5C aliphatic
glucosinolates was observed in BC3F2 progenies of the recurrent parent that carried the
GSL-ELONG- gene. Replacement of the functional locus in the A-genome by non-
functional counterpart in the C-genome reduced the content of 5C aliphatic
glucosinolates in B. rapa seeds with 20μmol/g.

29. A zero erucic acid C22:1 line of Brassica juncea VH486, adapted to the
agronomic conditions of Northern India, has been modified for its fatty acid
composition in the seed oil with antisense constructs using the sequence of
fad2 gene of B. rapa. Construct pASfad2.1 contained 315 to 1251 bp and
construct pASfad2.2 contained 1 to 1251 bp fragment of the fad2 gene, both in
antisense orientation, driven by a truncated napin promoter. Analysis of the
levels of linoleic acid C18:2 in the BC1 seeds of single-copy transgenic
showed that the construct pASfad2.2 gave better suppression of the fad2 gene
as compared to the construct pASfad2.1. These had ca. 73% C18:1 and 8 to
9% each of C18:2 and C18:3 -linolenic acid fractions in comparison to ca.
53% C18:1, 24% C18:2 and 16% C18:3 in the parental line VH486.

31.  From the review of this topic it can be inferred that zero erucic, low
linolenic, high oleic fatty acid and double zero genotypes are available and
some double zero varieties are also under cultivation in western countries.
There for more emphasis is required for improvement of oil and meal quality
in Rapeseed-mustard through extensive breeding work.
Erucic acid in oil and glucosinolate in cake are two nutritionally toxic
undesirable factors in rapeseed-mustard.
Back cross method is most widely used for quality
improvement in rapeseed-mustard
Conclusion

32.  Identification of diverse source of low glucosinolates, low erucic acid and
their utilization in the breeding programmes.
 Development of inexpensive and rapid method for mass screening of fatty
acids and glucosinolates to facilitate early generation selection.
 Use of biotechnological tools like somatic cell fusion, somaclonal variation,
marker-assisted breeding, and transformation in the development of plants
with novel desirable quality traits.
 Mapping and sequencing Brassica genomes will facilitate the isolation of
specific genes to aid in crop improvement.
FUTURE STRATEGIES

33. Thank
you