This document discusses efforts to introduce beneficial traits from wild relatives of pigeonpea into cultivated varieties. It describes the primary, secondary, tertiary, and quaternary gene pools of pigeonpea and traits discovered in secondary and tertiary gene pool species, including disease resistance from Cajanus acutifolius and a new A5 CMS system. It details the development of introgression lines with pod borer resistance from C. platycarpus and C. acutifolius using backcrossing and embryo rescue techniques.

1. Conquering gene pools of pigeonpea
[Cajanus cajan (L.) Millspaugh]
Nalini Mallikarjuna
Grain legumes Program
International Crops Research Institute for the
Semi-Arid Tropics, Patancheru, India.
ICRISAT is a member of the CGIAR Consortium

2. ICRISAT Collaborators
ICRISAT is a member of the CGIAR
Consortium

3. 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 base
ICRISAT is a member of the CGIAR
Consortium

4. How to tackle the brewing storm?
Water Scarcity and water loggingICRISAT is a member of the CGIAR Consortium

5. Gene pools of pigeonpea
Primary Gene Pool
Cajanus cajan and its land
races.
P
S
T
Q
Secondary Gene Pool
C. cajanifolius, C. lineatus, C. lanceolatus,
C. laticepalus, C. albicans, C. reticulatus,
C. sericeus, C. scarabaeoides, C .
trinervius, C. acutifolius,.
Tertiary Gene Pool
C. 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 Pool
Flemingia, 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

6. 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

7. 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

8. 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

9. Helicoverpa armigera damage to pigeonpea
ICRISAT is a member of the CGIAR Consortium

10. And the result is…!!!
ICRISAT is a member of the CGIAR Consortium

11. 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

12. Pod borer resistant Cajanus acutifolius
Fig.A total of 166 C.platycarpus and 301C.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

13. 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
Poddamage(%)
0
2
4
6
8
10
12
14
16
18
20
(%) Dam
100 seed wt
ICRISAT is a member of the CGIAR Consortium

14. 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

15. A5 CMS system
A5 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
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16. Cajanus cajan
Cajanus lanceolatus
Traits of interest transferred: A9 CMS
system, pod borer and bruchid resistance
INTER-SPECIFIC CROSS THAT IS FERTILE
Introgression of traits

17. 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.

18. 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

19. Helicoverpa resistant Cajanus platycarpus

20. 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 techniques
BC2
[87.5%A+12.5%B]
Screen for Helicoverpa
↓
[ May require growth regulator
applications, embryo rescue
techniques]
Interspecific
hybridization

21. 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

22. Molecular analysis
DArT 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.

23. Cajanus platycarpus derivatives are showing
variation for plant growth, height, seed
weight, disease and pest resistance
C. platycarpus seeds

24. Fig. A total of 166 C. platycarpus and 301C. 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

25. A7 CMS system
A7 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

26. Meiosis
Premeiotic
Postmeiotic

27. 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.

28. 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

29. Bruchid resistance
0
10
20
30
40
50
60
70
80
90
ICPW-
130
ICPW-66
ICPW-94
ICPW-
15671
ICPW-64
ICP-
15613
ICPW-68
Controls
85010
(Solidbars(%)seeddamage
0
10
20
30
40
50
60
70
80
90
Linegraphis(%)eggsunhatched
Fig. 4. Bruchid damage in w ild relatives of pigeonpea
0
20
40
60
80
100
237-1
238-1
A4-10--7--1
A4-10--7--2
A4-10--7--4
A4-10--7--7
A4-10-7--19
A4-10-7--20
7018--40-26-7-1
Cont.ICPL-
85010
Solidbars(%)seeddamage
0
20
40
60
80
100
Smallbars(%)eggdensity&
linegraphis(%)eggs
unhatched
Publication: Jadhav et al., 2012.

30. 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**

31. C. volubilis
First plant: Hybrid
Second plant: Cajanus cajan
Third plant: C. volubilis

32. SSR analysis of C. volubilis hybrid

33. Dwarf plant type and early maturity in F2
onwards
Srikanth et al., manuscript under preparation

34. Gene pools of pigeonpea
Primary Gene Pool
Cajanus cajan and its land
races.
P
S
T
Q
Secondary Gene Pool
C. cajanifolius, C. lineatus, C. lanceolatus,
C. laticepalus, C. albicans, C. reticulatus,
C. sericeus, C. scarabaeoides, C .
trinervius, C. acutifolius,.
Tertiary Gene Pool
C. 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 Pool
Flemingia, Rhynchosia,
Dunbaria, Erisema Paracalyx,
Adenodolichos, Bolusafra,
Carissoa, Chrysoscias, Baukea.
Fig. 1. Taken from Mallikarjuna et al., 2011

35. Quaternary gene pool
Recent Success :
Succeeded in crossing pigeonpea
with Rhynchosia species and
hybrids were obtained.

36. Crossing
pigeonpea
with
R. bracteata

37. SSR marker showing
polymorphism

38. Genomic in-situ hybridization

39. Summary
• Species from secondary, tertiary
and quarternary gene pool
crossed successfully with
pigeonpea

40. Summary
Diversification of CMS base
CMS 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)

41. 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.

42. Legume Cell Biology Staff
• Deepak R Jadhav
• V. Balakrishna
• S. Satyanarayaya
• K. Avinash
• K. Lakshmi
• A. Amrutha
and the students
Thank you Cell Biology staff for your help and
understanding and sharing the passion to pre-
breed ICRISAT mandate legumes.

43. End of the pigeonpea story from
me!!
Would you like to listen to the
grandoise groundnut
ground-breaking research??