Bett, K.

Vargas-Palacios, A.G.

English

University of Saskatchewan's Research Archive

Expanding the 
genetic resources of 
lentils and Rhizobium
to increase nitrogen 
fixation (BNF) in the 
lentil crop
Ana G. Vargas Palacios
Plant Science-University of Saskatchewan
March 6th,2018.
Outline
• Biological Nitrogen Fixation (BNF) in lentils
• Genetic Resources
Rhizobial strains Lentil genotypes/species
Methodology
Preliminary Results
• Future work
BNF in lentils
• Canada is the largest exporter/producer of lentils
• Symbiosis is the most important route to obtain N
- inoculation
• %Ndfa (Nitrogen derived from atmosphere)
- lentil: 0-87% (0-192 Kg/ha)
Modern varieties: usually selected under high fertility 
conditions.
FAO,2016. www.fao.org/faostat/en
Herridge et al., 2009. Plant and Soil 311:1-18.
Hypotheses
• The study of a broad group of cultivated and wild 
accessions within Lens species, will allow the 
identification of superior species/genotypes that 
can contribute positive N-fixation related alleles to 
modern lentil varieties.
• Exploring the genetic diversity of Rhizobium from 
the center of origin of lentils and other main 
production areas allows the incorporation of more 
efficient strains to the Saskatchewan cropping 
system, increasing N available in lentils. 
Genetic Resources
Harun-on-Rashid et al., 2014. FEMS Microbiology Ecology 87:64-77
Podder et al., 2012. Plant Genetic Resources 11:26-35
▪ L. culinaris, L. orientalis, L. tomentosus
▪ L. odemensis, L. lamottei
▪ L. ervoides
▪ L. nigricans
▪ R. leguminosarum bv. viciae
▪ R. bangladeshense
▪ R. lentils
▪ R. binae
▪ New species
Lentils
Rhizobium strains
Methodology: strains
Riely et al., 2017. UC Davis
14 strains <-> CDC Maxim
BNF potential under local    
conditions
Controls: BASF 4035, Non 
inoculated
4 site-years: Sutherland, 
Rosthern (1-2017, 3-2018)
Soil analysis record for N content
RCBD: 8 repetitions
15 Rhizobium treatments
120 plots
Methodology: strains
OD measurements of 15 strains at 600 nm during 84 hours
Survival on 
seed 
surface
Seed 
inoculation and 
drying
Methodology: strains
At flowering (10 plants center row): 
Shoot area: N content, SDW (g)
Roots
-Presence and structure of nodules: 
number, position, color
-NDW
At maturity (10 plants+6 rows)
-yield traits: #pots/plant, 1000 SW
-yield
-Protein (seed)
Photos: Karsten Nielsen
Preliminary Results
OyaliB
NZLR1
GL17
Nodulation in 
the field
A
B
C
Strain Species Yield(Kg/ha) Protein (%)
NZLR24 R. leguminosarum 2587a 25.90a
NZLR1 R. leguminosarum 2409  b 25.38a
ALM1 R. leguminosarum 2377  bc 25.52a
BASF 4035 R. leguminosarum 2339  bcd 25.47a
GLR54 R. leguminosarum 2335  bcd 25.32a
PLR8_1a R. bangladeshense 2333  bcd 24.51  b
Non-inoculated 2328  bcd 25.21ab
LEN3 R. leguminosarum 2307  bcde 25.61a
TLR11 R. leguminosarum 2275  bcde 25.64a
GL13 R. leguminosarum 2248  bcde 25.38a
BL27(T) R. lentils 2245  bcde 25.55a
OyaliB R. leguminosarum 2214    cde 25.44a
GL17 R. leguminosarum 2211    cde 25.49a
BL195(T) R. binae 2180      de 25.57a
GLR28 R. leguminosarum 2145        e 25.77a
LSD,CV                                                                         174.9, 7.66                      0.74,2.94
Yield of CDC Maxim inoculated with 15 different Rhizobium strains. Sutherland, SK. 2017
Methodology: genotypes
36 genotypes 
7 Lens species 
Sand:Sunshine (1:1)
pasteurized 
N free nutrient solution 
Seed 
scarification/disinfection 
Inoculum 
BASF 4035-YMB media
Quality control
1×109 cells/ml
Inoculation
1 ml/plant
Somasegaran and Hoben. 1991. Handbook for Rizobia
+N     Rhizobium   -N
Methodology: genotypes
Split plot design (432 exp.units)
Methodology: genotypes
Evaluation:
Photosynthesis related parameters 
MultispepQ with PhotosynQ app
Chlorophyll Fluorescence
Relative Chlorophyll: SPAD
Leaf Temperature and differential from 
ambient temperature
At flowering: 
Shoot area: N content, SDW (g)
Roots
-Presence and structure of nodules: 
number, position, color
-Efficiency (N2fixed/nodule)
Nodulation in the GH
L. orientalis
IG 72611
L. culinaris
CDC Maxim
L. orientalis
IG 72643
Shoot and root dry weight of 36 genotypes of lentils 
inoculated with R. leguminosarum
Relative Chlorophyll and Nitrogen of 36 genotypes of lentils 
inoculated with R. leguminosarum
Number of nodules of 36 genotypes of lentils inoculated 
with R. leguminosarum
Nitrogen fixed (mg)/nodule of 36 genotypes of lentils 
inoculated with R. leguminosarum
Image-based analyses
Study of interspecific populations 
Genetic mapping of QTL’s associated with higher BNF ability 
in interspecific populations
What’s next?
Kirstin Bett
Bert Vandenberg
Diane Knight
Brendan Riely
Douglas Cook
Brent Barlow
Devini Da Silva
Eldon Siemens
Linda Gorim
Crystal Chan
Robert Stonehouse
Akiko Tomita
Karsten Nielsen
Pulse Field Crew


Expanding the genetic resources of lentils and Rhizobium to increase nitrogen fixation (BNF) in the lentil crop

https://harvest.usask.ca/bitstream/10388/8668/1/A.%20Palacios%20and%20K.%20Bett%2c%202018.pdf

Expanding the genetic resources of lentils and Rhizobium to increase nitrogen fixation (BNF) in the lentil crop

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