Conquering gene pools of pigeonpea [Cajanus cajan (L.) Millspaugh]
Nalini Mallikarjuna
Grain legumes ProgramInternational Crops Research Institute for the
Semi-Arid Tropics, Patancheru, India.
ICRISAT is a member of the CGIAR Consortium
ICRISAT Collaborators
ICRISAT is a member of the CGIAR Consortium
Pigeonpea Domestication Bottleneck/s
One founder species i.e. Cajanus cajanifolius
Indian origin
Diploid cultivated pigeonpea
Farmer’s selection
Plant breeding
Pigeonpea with a narrow genetic baseICRISAT is a member of the CGIAR Consortium
How to tackle the brewing storm?
Water Scarcity and water loggingICRISAT is a member of the CGIAR Consortium
Gene pools of pigeonpeaPrimary Gene PoolCajanus cajan and its land races.
P
S
T
Q
Secondary Gene PoolC. cajanifolius, C. lineatus, C. lanceolatus, C. laticepalus, C. albicans, C. reticulatus, C. sericeus, C. scarabaeoides, C . trinervius, C. acutifolius,.
Tertiary Gene PoolC. goensis, C. heynei, C. kerstingii, C. mollis,C. rugosus, C. volubilis, C. platycarpus, C. niveus, C. gandiflorus, C. crassicaulis, C. rugosus, C. elongates, C. villosus, C . confertiflorus, C. visidus, C. aromaticus, C. crassicaulis, C. lanuginosus, C. pubescens, C. cinereus, C. marmoratus, C. mareebensis.C. lanuginosus, C. pubescens.
Quaternary gene PoolFlemingia, Rhynchosia, Dunbaria, Erisema Paracalyx, Adenodolichos, Bolusafra, Carissoa, Chrysoscias, Baukea.
Fig. 1. Taken from Mallikarjuna et al., 2011
ICRISAT is a member of the CGIAR Consortium
Primary gene pool:
• >13,600 accessions
• Sixty-six cultivars based on germplasm and improved breeding lines developed by ICRISAT have been released in several countries of Asia (38), Africa (22), Australia (3) and USA (3). Short and medium duration cultivars have made a significant impact in Asian countries. However, long and medium duration varieties with fusarium wilt resistance and consumer preferred large, cream colored seeds have created impacts in eastern and southern Africa.
• Variation for days to flower, maturity, yield, harvest index, shelling ratio with limited amount of disease and pest resistance etc.
• High yielding varietiese and useful genetic stocks
ICRISAT is a member of the CGIAR Consortium
Secondary gene pool
•Compatible wild relatives
•Traits discovered:
• C. acutifolius: A5 CMS system, pod borer, pod fly, bruchid resistance, phytophthora blight, dwarf plant type, high seed weight, beige seed color.
• C. lanceolatus: A9 CMS system, bruchid resistance.
ICRISAT is a member of the CGIAR Consortium
Cajanus acutifolius
• A wild relative from secondary gene
pool.
• Native of Auatralia
• Has a repertoire of disease and
pest resistance traits.
• One way cross more successful
than the reciprocal.
ICRISAT is a member of the CGIAR Consortium
Helicoverpa armigera damage to pigeonpea
ICRISAT is a member of the CGIAR Consortium
And the result is…!!!
ICRISAT is a member of the CGIAR Consortium
How does this happen?
Happy Helicoverpa but sad peas!!!!
Publications: Mallikarjuna et al., 2007, Mallikarjuna et al., 2012a, Mallikarjuna et al., 2012b
ICRISAT is a member of the CGIAR Consortium
Pod borer resistant Cajanus acutifolius
Fig. A total of 166 C. platycarpus and 301 C. acutifolius interspecific derivative lines with low Helicoverpa armigera damage
0
50
100
150
200
250
300
11-10 11-20 20-30 30-40
Helicoverpa pod damage
C. platycarpus
C. acutifolius
ICRISAT is a member of the CGIAR Consortium
High seed weight lines
Fig. Helicoverpa armigera pod damage and high 100 seed wt. in a Cajanus acutifolius derived advanced progeny line
0
10
20
30
40
50
60
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 R-cont-icp-332
S-cont-
87
progenies
Pod
dam
age
(%)
0
2
4
6
8
10
12
14
16
18
20
(%) Dam
100 seed wt
ICRISAT is a member of the CGIAR Consortium
C. acutifolius lines with multiple disease resistance
ICPLNos FW SM PB SWT
ICPL2011146 12.5** 0** 8** 13.6**
ICPL2011147 11.1** 0** 2** 11.6**
ICPL2011148 0** 0** 1** 17.3**
ICPL2011150 0** 0** 2** 12.4**
ICPL2011160 14.3** 0** 2** 18.4**
ICPL2011164 12.5** 0** 7** 20**
ICPL2011168 14.3** 0** 3** 19.6**
ICPL2011169 16.7** 0** 3** 19.6**
ICPL2011183 11.1** 0** 2** 18.8**
ICPL2011170 11.1** 0** 3** 19.2**
ICPL2011161 0** 0** 2** 18.8**
ICPL2011203 12.5** 0** 1** 16.9**
Publications: Jadhav et al., 2012 a & b, Mallikarjuna et al., manuscripts under preparation
ICRISAT is a member of the CGIAR Consortium
A5 CMS systemA5 CMS system was developed utilizing C. acutifolius and with cultivated pigeonpea cytoplasm.
•First report by Mallikarjuna and Saxena (2005).
•The only system with cultivated pigeonpea cytoplasm
•Has post meiotic pollen abortion
•Plenty of restorers available
•Maintainers have been identified
•Handed it over to the breeders
ICRISAT is a member of the CGIAR Consortium
Cajanus cajan
Cajanus lanceolatus
Traits of interest transferred: A9 CMS system, pod borer and bruchid resistance
INTER-SPECIFIC CROSS THAT IS FERTILEIntrogression of traits
A9 CMS system
• A9 CMS system was recently discovered
by us.
• The beauty of the system is that it is developed on cultivated pigeonpea cytoplasm.
• It was possible to identify maintainers and restorers in pigeonpea cultivars
• Will be handed over to breeders shortly.
Tertiary gene pool
•Incompatible Cajanus species
•C. platycarpus: A7CMS, clasmogamy,pod borer, pod-fly, bruchid and Phytophthora blight resistance.
•C.volubilis: dwarf plant type, early maturity and high pod number.
ICRISAT is a member of the CGIAR Consortium
Helicoverpa resistant Cajanus platycarpus
Compatible species Incompatible species
Cultivated species wild species[100% A] [100% B]
F1
[50%A+50%B]
BC1
[75%A+25%B]Screen for Helicoverpa
BC2
[87.5%A+12.5%B]Screen for Helicoverpa
BC3
[93.75%A+6.25%B]Screen for Helicoverpa
Cultivated species Wild species[100%A] [100%B]
[or vice versa]
Growth regulator applications
Embryo rescue techniques F1
[50%A+50%B]
Growth regulator applications
Embryo rescue techniques BC1
[75%A+25%B]Screen for Helicoverpa
Growth regulator applications,
Embryo rescue techniquesBC2
[87.5%A+12.5%B]Screen for Helicoverpa
[ May require growth regulator
applications, embryo rescue techniques]
BC3
[93.75%A+ 6.25%B]Screen for Helicoverpa
Interspecific hybridization
Mature seed set from BC2
onwards
Embryo abortion in BC1
F
Embryo abortion in F1
FM
Embryo rescue in recurrent backcrosses
Publications: Mallikarjuna et al, 2007, Mallikarjuna et al., 2011; Mallikarjuna et al., 2012
F
Molecular analysisDArT analysis showed the presence of C. platycarpus genome ranging from 2.0 to 4.8%. The presence of non-parental alleles, presumably due to recombination, ranged from 2.6 to 10.4%. These two categories have contributed to the variation observed in the progenies.
Cajanus platycarpus derivatives are showing variation for plant growth, height, seed
weight, disease and pest resistance
C. platycarpus seeds
Fig. A total of 166 C. platycarpus and 301 C. acutifolius interspecific derivative lines with low Helicoverpa armigera damage
0
50
100
150
200
250
300
11-10 11-20 20-30 30-40
Helicoverpa pod damage
C. platycarpus
C. acutifolius
A7 CMS systemA7 CMS system was developed utilizing C. platycarpus, a species from tertiary gene pool and incompatible with pigeonpea
• First reported by Mallikarjuna et al., 2011
• Has both pre-meiotic and post- meiotic pollen abortion.
• Handed over to pigeonpea breeders in 2012
Meiosis
Premeiotic
Postmeiotic
Important traits of A7 CMS system
• Clasmogamous flowers
• Pre and post meiotic anther abortion
• Non-dehiscent anthers
Publications: Cherian et al., 2007, Mallikarjuna et al., 2012.
Clasmogamous or open flower is an important trait in the development of CMS systems in pigeonpea. A7 CMS system had open flowers accompanied by sterile and non-dehiscent anthers, an important trait in this CMS system
Bruchid resistance
0102030405060708090
ICP
W-
13
0
ICP
W-6
6
ICP
W-9
4
ICP
W-
15
67
1
ICP
W-6
4
ICP
-1
56
13
ICP
W-6
8
Co
ntr
ols
85
01
0
(So
lid b
ars
(%
) se
ed
da
ma
ge
0102030405060708090
Lin
e g
rap
h is
(%
) e
gg
s u
nh
atc
he
d
Fig. 4. Bruchid damage in w ild relatives of pigeonpea
020406080
100
237-
1
238-
1
A4-1
0--7
--1
A4-1
0--7
--2
A4-1
0--7
--4
A4-1
0--7
--7
A4-1
0-7-
-19
A4-1
0-7-
-20
7018
--40
-26-
7-1
Con
t. IC
PL-
8501
0 Solid
bar
s (%
) see
d da
mag
e
020406080100
Smal
l bar
s (%
) egg
den
sity
&
line
grap
h is
(%) e
ggs
unha
tche
d
Publication: Jadhav et al., 2012.
C. platycarpus lines with multi disease resistance ICPL NO FW SM PB SWT
ASHA 40.8 60.64 31.3 9.67
ICPL2011007 100.0** 0.00** 1.00** 8.2**
ICPL2011013 100.0** 0.00** 1.00** 7.7**
ICPL2011014 100.0** 0.00** 1.00** 7.3**
ICPL2011015 100.00** 0.00** 1.00** 8.2**
ICPL2011016 83.3** 16.7** 1.00** 7.6**
ICPL2011018 100.0** 0.00** 3.00** 7.6**
ICPL2011024 0.00** 100.0** 2.00** 12.0**
ICPL2011025 100.0** 0.00** 3.00** 8.7**
ICPL2011029 100.0** 0.00** 3.00** 8.7**
ICPL2011033 85.7** 0.00** 2.00** 8.4**
ICPL2011037 88.9** 0.00** 4.00** 8.7**
ICPL2011041 83.3** 16.7** 3.00** 8.5**
ICPL2011044 100.0** 0.00** 7.00** 7.8**
ICPL2011047 100.0** 0.00** 3.00** 11.1**
ICPL2011061 100.0** 0.00** 2.00** 8.1**
ICPL2011072 0.00** 100.0** 3.00** 8.6**
ICPL2011074 0.00** 0.00** 5.00** 7.8**
ICPL2011088 88.9** 0.00** 2.00** 6.6**
ICPL2011090 100.0** 0.00** 2.00** 7.8**
ICPL2011124 75.0** 12.5** 1.00** 8.2**
ICPL2011127 100.0** 0.00** 2.00** 7.3**
ICPL2011128 75.0** 0.00** 4.00** 8.00**
ICPL2011129 100.0** 0.00** 4.00** 7.9**
ICPL2011132 100.0** 0.00** 7.0** 12.4**
ICPL2011133 100.0** 0.00** 2.0** 8**
C. volubilis
First plant: Hybrid
Second plant: Cajanus cajan
Third plant: C. volubilis
SSR analysis of C. volubilis hybrid
Dwarf plant type and early maturity in F2 onwards
Srikanth et al., manuscript under preparation
Gene pools of pigeonpeaPrimary Gene PoolCajanus cajan and its land races.
P
S
T
Q
Secondary Gene PoolC. cajanifolius, C. lineatus, C. lanceolatus, C. laticepalus, C. albicans, C. reticulatus, C. sericeus, C. scarabaeoides, C . trinervius, C. acutifolius,.
Tertiary Gene PoolC. goensis, C. heynei, C. kerstingii, C. mollis,C. rugosus, C. volubilis, C. platycarpus, C. niveus, C. gandiflorus, C. crassicaulis, C. rugosus, C. elongates, C. villosus, C . confertiflorus, C. visidus, C. aromaticus, C. crassicaulis, C. lanuginosus, C. pubescens, C. cinereus, C. marmoratus, C. mareebensis.C. lanuginosus, C. pubescens.
Quaternary gene PoolFlemingia, Rhynchosia, Dunbaria, Erisema Paracalyx, Adenodolichos, Bolusafra, Carissoa, Chrysoscias, Baukea.
Fig. 1. Taken from Mallikarjuna et al., 2011
Quaternary gene poolRecent Success :Succeeded in crossing pigeonpea with Rhynchosia species and hybrids were obtained.
Crossing
pigeonpea
with
R. bracteata
SSR marker showing polymorphism
Genomic in-situ hybridization
Summary
• Species from secondary, tertiary and quarternary gene pool crossed successfully with pigeonpea
SummaryDiversification of CMS baseCMS system Wild species
A1 C. sericeus
A2 C. scarabaeoides
A3 C. volubilis
A4 C. cajanifolius
A5 C. acutifolius (Mallikarjuna & Saxena, 2005)
A6 C. lineatus
A7 C. platycarpus (Mallikarjuna et al., 2011)
A8 C. reticulatus
A9 C. lanceolatus (Srikanth et al, manuscript under preparation)
Summary
• Disease and pest resistant stable pre-breeding lines obtained from secondary gene pool species.
• Successfully crossed tertiary gene pool species and developed stable disease and pest pre-breeding lines.
• Dwarf and early plant type.
• High seed weight and number.
Legume Cell Biology Staff
• Deepak R Jadhav• V. Balakrishna• S. Satyanarayaya• K. Avinash• K. Lakshmi• A. Amruthaand the students Thank you Cell Biology staff for your help and
understanding and sharing the passion to pre-breed ICRISAT mandate legumes.
End of the pigeonpea story from me!!
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ground-breaking research??
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