Performance of grass pea (Lathyrus sativus L.) somaclones at Adet, northwest Ethiopia

37 Pages·2006·1.52 MB·English

by  Colin Hanbury

Lathyrus Lathyrism Newsletter 4 (2005)

Contents

Page
Editor's Comment
1 Colin Hanbury -Australia
Articles
2 Fatty acid composition of grass pea (Lathyrus sativus L.) Gurusamy Chinnasamy, Arya Kumar Bal,
seeds and David Bruce McKenzie- Canada
5 Performance of grass pea (Lathyrus sativus L.) somaclones D. Tsegaye, W. Tadesse and M. Bayable-
at Adet, northwest Ethiopia Ethiopia
7 Search for resistance to crenate broomrape (Orobanche J.C. Sillero, J.I. Cubero, M. Fernández-
crenata) in Lathyrus Aparicio and D. Rubiales- Spain
10 Characterization of grass pea (Lathyrus sativus L.) entries G. B. Polignano, P. Uggenti, G. Olita, V.
by means of agronomically useful traits Bisignano, V. Alba and P. Perrino- Italy
15 Model plant type in Khesari (Lathyrus sativus L.) suitable Vedna Kumari and Rajendra Prasad- India
for hill farming
18 Resilience of South Asian disabling conditions: a glimpse M. Miles- UK
of lathyrism among comparative histories
22 Considerations on the reintroduction of grass pea in China Hui-Min Yang and Xiao-Yan Zhang- China
27 Effects of drought on stomatal character, photosynthetic Hui-Min Yang, Xiao-Yan Zhang and Gen-
character and seed chemical composition in grass pea, and Xuan Wang- China
their relationships
28 Scope of growing lathyrus and lentil in relay cropping S. Gupta and M.K. Bhowmick- India
systems after rice in West Bengal, India
34 The same goal, a different approach: a new Belgian- Fernand Lambein and Seid Ahmed-
Ethiopian project Belgium/Ethiopia
THIS VOLUME IS STILL BEING COMPILED- THERE WILL BE MORE ARTICLES TO FOLLOW

ISSN 1832-8431 (Print) and ISSN 1832-844X (Online)

The Lathyrus Lathyrism Newsletter can be obtained on-line at

http://go.to/lathyrus
OR
http://www.clima.uwa.edu.au/lathyrus

All research articles are provided there in PDF format.
Lathyrus Lathyrism Newsletter 4 (2005)

Jointly supported by:
Third World Medical Research Foundation (TWMRF),
PO Box 9171, Portland, Oregon 97207, USA
http://www.twmrf.org
and
Centre for Legumes in Mediterranean Agriculture (CLIMA),
University of Western Australia, 35 Stirling Highway
Crawley 6009, Australia
http://www.clima.uwa.edu.au
Lathyrus Lathyrism Newsletter 4 (2005)

Editor’s comment

Dear Readers

Welcome to the Lathyrus Lathyrism Newsletter Vol. 4 and thank you all for your support of the newsletter. You will
see from the variety of articles that there is enthusiasm for Lathyrus related research in many countries, this bodes
well for the increased knowledge and cultivation of these useful legumes.

Recently, there has been a change in the editing process. In order to give authors an advantage of on-line publication
from now onward articles will be placed on the web page as soon as the editing process is finalised. This will enable
the maximum quick exposure of information. Then at the end of the year the volume will be closed off and compiled,
for those who require the printed version this is when it will be completed. Hence as you revisit the web page through
the year you will notice more articles appearing. Any suggestions for further improvements are welcome.

The Lathyrus Lathyrism Newsletter has been assigned ISSN 1832-8431 (Print) and ISSN 1832-844X (Online) and has
been placed on a number of directories of open access journals, ensuring wider exposure and listings in library
resources internationally. Citations of articles in the newsletter have been steadily increasing.

Thanks to the Third World Medical Research Foundation (TWMRF) and the Centre for Legumes in Mediterranean
Agriculture (CLIMA) for supporting the newsletter from 2000 until now.

Please consider summarising your recent research for the newsletter. Your contribution will be useful not only as
science but also toward the greater stability of cropping systems in regions with increasing pressure on natural
resources. Most research submissions should be approximately 1500 words and can include a small number of tables
or figures, with electronic copies preferred. Introduction, Methods, Results followed by Discussion is the preferred
layout for research summaries, although this can be altered as necessary. Abstracts of complete work are also
welcome, if they have been published elsewhere then full acknowledgment will be made.

I hope you find the articles contained interesting and useful to your work.

Colin Hanbury

Editor contact details:

Dr Colin Hanbury
Department of Agriculture, Western Australia
3 Baron-Hay Court
South Perth 6151
Australia

E-mail: [email protected]

1
Lathyrus Lathyrism Newsletter 4 (2005)

Fatty acid composition of grass pea (Lathyrus sativus L.) seeds

Gurusamy Chinnasamy1*, Arya Kumar Bal1, and David Bruce McKenzie2

1. Department of Biology, Memorial University of Newfoundland, St. John’s, Newfoundland and Labrador,
Canada A1B 3X9.
2. Atlantic Cool Climate Crop Research Centre, Agriculture and Agri-Food Canada, 308 Brookfield Road, St.
John’s, Newfoundland and Labrador, Canada A1E 5Y7.

*Author for correspondence. Present address: Imaging Program, Lawson Health Research Institute, St.
Joseph’s Health Care, 268 Grosvenor Street, London, Ontario, Canada N6A 4V2
Email: [email protected] OR [email protected]

Introduction represent a potential source of several important
Grass pea or chickling vetch (Lathyrus sativus L.) is a nutrients for human and animal nutrition. Therefore, it
well-established, commercially available, tropical is necessary to analyze grass pea seeds for their
semi-arid crop. Pods of this crop are flat, dorsally nutrient composition. The present investigation
broad with two ridges, short and 3-5 cm in length. proposes to determine the fatty acid composition of
Each pod contains 2-7 seeds. Mature seeds are different lipid classes in mature seeds of Indian grass
rhomboid or triangular in shape, dull whitish grey pea.
brown and variously mottled (4). Grass pea is a
relatively productive crop compared to other pulses in Material and Methods
regions characterized by poor soil (7). It is very well Seed materials
adapted to adverse climatic conditions and requires Mature seeds of grass pea were procured from a local
very little management for crop production. Moreover, market in Kolkatta (Calcutta), West Bengal, India.
its deep taproot and nitrogen-fixing ability make this
crop an ideal choice for sustainable agriculture. Grass Extraction and estimation of total lipids
pea seeds are a major source of protein for large Total lipids were determined by the gravimetric
sections of the population in Bangladesh, China, method (2). One gram of dried grass pea seeds in each
Ethiopia and India (13, 15). It is also grown to a lesser of three replicates was powdered using a 700S Waring
extent in the Middle East, southern Europe and some blender (Waring Products Co., USA) and
parts of South America. In India grass pea is homogenized in 10 ml of 50 mM Tris-HCl buffer
considered one of the most economical pulses for containing 0.5 M NaCl at pH 7.2. The homogenate
fodder and green manure in rice fields during the cool was combined with a mixture of chloroform and
winter period (1). methanol in a ratio of 1.25:2.25 (v/v) to extract lipids.
The chloroform layer (supernatant) was separated by
Lathyrus species contain very high protein, but a centrifugation at 5000 g for 20 min and allowed to
neurotoxin, 3-(-N-oxalyl)-L-2,3-diamino propionic stand overnight after combining with a mixture of
acid (ODAP), is present in wild and most cultivated chloroform and distilled water in a ratio of 1:1 (v/v).
forms that if consumed in sufficient amounts can The chloroform layer was collected in a pre-weighed
cause the irreversible crippling disease known as vial for evaporation under nitrogen gas. The vial with
lathyrism (10, 17). This toxin to a considerable extent the lipid residue was weighed again to estimate the
has been bred out of some cultivars although lathyrism amount of total lipids.
in Asia from consuming grass pea is common.
Because of its drought tolerance, grass pea has been Separation of lipids and analysis of fatty acid
judged to have good potential as a future new pulse composition
crop for low rainfall areas of the Canadian prairies (12). Total lipids were fractionated into 5 lipid classes
It acts as a ground cover alternative to summer fallow, [phospholipids (PL), monoglycerides (MG),
helping to prevent wind and water erosion, as well as diglycerides (DG), free fatty acids (FFA) and
adding nitrogen to the soil (11). triglycerides (TG)] by thin layer chromatography and
the constituent fatty acids in each lipid class were
The nutritional health and well being of humans are separated and estimated using gas chromatography as
entirely dependent on plant foods. Plants are critical described in Chinnasamy et al. (6). The measurement
components of the dietary food chain in that they of each fatty acid was calculated as a relative weight
provide almost all essential minerals and organic percentage to 10 selected fatty acids [C14:0 (myristic
nutrients to humans either directly, or indirectly when acid), C14:1 (myristoleic acid), C16:0 (palmitic acid),
plants are consumed by animals, which are then C16:1 (palmitoleic acid), C18:0 (stearic acid), C18:1
consumed by humans (9). Grass pea seeds may (oleic acid), C18:2 (linoleic acid), C18:3 (linolenic
2
Lathyrus Lathyrism Newsletter 4 (2005)

acid), C18:4 and C20:4 (arachidonic acid)]. The from C8 to C22 in total lipids of Canadian grass pea
double bond index (DBI) was calculated (14) using the seeds. The relative weight percentages of 10 major
formula: fatty acids in 5 lipid classes isolated from mature
seeds of Indian grass pea are summarized in Table 1.
DBI=Σ (% of fatty acid content × no. of double bonds)
100 C18:2, C18:1, C18:0 and C16:0 were the major fatty
acids present in PL, MG, FFA and TG separated from
Among 10 major fatty acids, sum of all saturated fatty grass pea seeds. DG contained higher quantities of
acids, unsaturated fatty acids with one double bond C18:0, C16:0 and C18:2 compared to other fatty acids.
and unsaturated fatty acids with more than one double PL and TG showed higher overall DBI than FFA, MG
bond gave total saturated fatty acids (TSFA), total and DG. The content of TSFA was higher in DG than
monounsaturated fatty acids (TMUFA) and total in other lipid classes. High amounts of TMUFA were
polyunsaturated fatty acids (TPUFA) respectively. observed in FFA and TG. PL registered high quantity
Unsaturated to saturated ratio (USR) was calculated of TPUFA. TG, PL and FFA showed higher USR
by dividing total unsaturated fatty acids by total compared to MG and DG. Although lipids constitute a
saturated fatty acids. minor portion of many leguminous seeds, their
profiles indicate the desirable nature of fatty acid
Statistical analysis constituents present (3). In the present study, grass pea
For all sets of data, one way analysis of variance was seeds exhibited a high amount of total unsaturated
performed using the SPSS computer package (16). fatty acids (56.37% – 59.98%) and a low amount of
Means were compared by Duncan’s multiple total saturated fatty acids (40.01 – 43.65%) in all lipid
comparison test at P = 0.05. For the purpose of classes except MG and DG, which contained 31.49 –
statistical analysis, data in percentage were 47.29 % total unsaturated fatty acids and 52.72 –
transformed to arcsine values (18). 68.53% total saturated fatty acids. Overall, in the
present work, Indian grass pea seeds showed higher
Results and Discussion total unsaturated fatty acids than total saturated fatty
Total lipid content of mature grass pea seeds was acids that are in agreement with fatty acid composition
20.93 ± 0.27 mg/g dry weight. The nutritive value of of beach pea and Canadian grass pea seeds (3-5).
seeds is determined by not only quantity but also by Therefore, grass pea seeds may be important for
quality of lipids they contain. Thus, fatty acids present nutritional health and may serve as a valuable
in lipids are playing important role in deciding shelf nutritional source. Further in depth study is necessary
life, nutrition and flavor of food products (8). Chavan to elucidate the nutritional quality and importance of
et al. (3) reported the presence of 20 fatty acids varying grass pea.

Table 1. Composition of major fatty acids (relative weight percentage1) in phospholipids (PL), monoglycerides
(MG), diglycerides (DG), free fatty acids (FFA) and triglycerides (TG) isolated from total lipids of mature
seeds of Indian grass pea. Values are means (± SE) of three replications.

Fatty acids PL MG DG FFA TG
C14:0 0.65 ± 0.82b 4.51 ± 2.78b 2.01 ± 0.24d 2.09 ± 1.61c 1.06 ± 0.85c
C14:1 0.66 ± 0.53b 0.77 ± 1.41b 0.91 ± 1.63de 0.49 ± 0.09c 0.82 ± 1.03c
C16:0 24.88 ± 1.67a 26.62 ± 2.89a 31.21 ± 0.75a 26.07 ± 3.60ab 24.57 ± 5.25ab
C16:1 0.57 ± 0.40b 2.77 ± 2.53b 1.10 ± 0.48de 2.66 ± 2.61c 2.26 ± 1.17c
C18:0 16.24 ± 4.90a 21.59 ± 1.82a 35.31 ± 0.92a 15.49 ± 1.15b 14.38 ± 1.96b
C18:1 26.20 ± 6.64a 23.44 ± 2.18a 9.30 ± 0.12c 35.99 ± 8.32a 33.91 ± 6.64a
C18:2 28.65 ± 11.13a 17.84 ± 8.39a 17.55 ± 1.86b 14.31 ± 2.85b 19.17 ± 3.73ab
C18:3 1.16 ± 0.61b 1.57 ± 2.27b 0.75 ± 0.69de 0.81 ± 0.26c 2.55 ± 1.90c
C18:4 0.64 ± 0.91b 0.23 ± 0.19b 0.73 ± 0.99e 1.93 ± 1.61c 0.75 ± 1.35c
C20:4 0.36 ± 1.32b 0.67 ± 0.70b 1.15 ± 0.89de 0.18 ± 0.04c 0.52 ± 1.03c
DBI2 0.92 ± 0.18 0.71 ± 0.20 0.56 ± 0.08 0.79 ± 0.02 0.88 ± 0.04
TSFA3 41.77 52.72 68.53 43.65 40.01
TMUFA4 27.43 26.98 11.31 39.14 36.99
TPUFA5 30.81 20.31 20.18 17.23 22.99
USR6 1.39 0.90 0.46 1.29 1.50

1The value of each fatty acid was calculated as a relative a-e Means in the same column followed by different letters
weight percentage to 10 selected fatty acids. are significantly different using Duncan’s multiple
2 Double bond index. comparison test at P = 0.05.
3 Total saturated fatty acids.
4 Total monounsaturated fatty acids.
5 Total polyunsaturated fatty acids.
6 Unsaturated/saturated ratio.
3
Lathyrus Lathyrism Newsletter 4 (2005)

Acknowledgements
We wish to thank Dr. P. J. Davis, Rhonda and Dona
for the help in fatty acid analysis. This work was
supported by the Dean of Science Grant to A. K. Bal.

References
1. Allen ON, Allen EK. 1981. The leguminosae. 10. Hanbury C, White C, Mullan BP, Siddique KHM.
University of Wisconsin Press, Madison, WI. 2000. A review of the potential of Lathyrus
2. Bligh EG, Dyer WJ. 1959. A rapid method of sativus L. and L. cicera L. grain for use as animal
total lipid extraction and purification. Can J feed. Lathyrus Lathyrism Newsletter 1, 34.
Biochem Physiol 37, 911-917. 11. Henkes R. 1995. The remaking of grasspea. The
3. Chavan UD, Shahidi F, Bal AK, McKenzie DB. Furrow 100, 25-26.
1999. Physico-chemical properties and nutrient 12. Kiehn FA, Reimer M. 1992. Alternative crops for
composition of beach pea (Lathyrus maritimus the prairies. Agriculture Canada Publication.
L.). Food Chem 66, 43-50. 1887/E, Ottawa, Ontario, Canada.
4. Chavan UD. 1998. Chemical and biochemical 13. Kuo YH, Khan JK, Lambein F. 1994.
components of beach pea (Lathyrus maritimus Biosynthesis of the neurotoxin β-ODAP in
L.). Ph.D. Thesis. Memorial University of developing seeds of Lathyrus sativus. Phytochem
Newfoundland, St. John’s, Newfoundland and 35, 911-913.
Labrador, Canada. 14. Skoczowski A, Filek M, Dubert F. 1994. The
5. Chinnasamy G, Bal AK, McKenzie DB. 2004. long-term effect of cold on the metabolism of
Fatty acid and elemental composition of mature winter wheat seedlings. II. Composition of fatty
seeds of beach pea [Lathyrus maritimus (L.) acids of phospholipids. J Therm Biol 19, 171-176.
Bigel.]. Can J Plant Sci 84, 65-69. 15. Spencer PS, Roy DN, Ludolph A, Dwivedi MP,
6. Chinnasamy G, Davis PJ, Bal AK. 2003. Seasonal Roy DN, Hugon J, Schaumburg HH. 1986.
changes in oleosomic lipids and fatty acids of Lathyrism: evidence for the role of the
perennial root nodules of beach pea. J Plant neuroexcitatory amino acid BOAA. Lancet 2,
Physiol 160, 355-365. 1066-1067.
7. Duke JA, Reed CF, Weder JKP. 1981. Lathyrus 16. SPSS Inc. 1990. SPSS/PC + StatisticsTM 4.0 for
sativus L. In Handbook of legumes of world the IBM PC/XT/AT and PS/2. SPSS Inc.,
economic importance. Duke JA (Ed.). Plenum Chicago, IL.
Press, New York, NY. pp. 107-110 and 345-349. 17. White C, Hanbury C, Siddique KHM. 2001. The
8. Gaydou EM, Rasoarahona J, Bianchini JP. 1983. nutritional value of Lathyrus cicera and Lupinus
A micromethod for the estimation of oil content angustifolius grain for sheep. Lathyrus Lathyrism
and fatty acid composition in seeds with special Newsletter 2, 49-50.
reference to cyclopropenoic acids. J Sci Food 18. Zar JH. 1996. Biostatistical analysis. Third
Agric 34, 1130-1136. edition. Prentice Hall, New Jersey, NJ.
9. Grusak MA, DellaPenna D. 1999. Improving the
nutrient composition of plants to enhance human
nutrition and health. Ann Rev Plant Physiol Plant
Mol Biol 50, 133-161.

4
Lathyrus Lathyrism Newsletter 4 (2005)

Performance of grass pea (Lathyrus sativus L.) somaclones
at Adet, northwest Ethiopia

D. Tsegaye*, W. Tadesse and M. Bayable

Adet Agricultural Research Center, P.O Box 08, Bahir Dar, Ethiopia.

*Email: [email protected]

Introduction Material and Methods
Grass pea (Lathyrus sativus L.) is one of the important Eleven low neurotoxin grass pea somaclones
crops of economic significance in Ethiopia. It is the introduced from ICARDA and a local check were
fifth most important pulse crop in Ethiopia after faba grown for two consecutive years (2000/01 and
bean, field pea, chickpea and haricot bean. It is the 2001/02) at Adet Agricultural Research Center, which
cheapest source of protein in the diets of most people. is located at 37029’E and 11016’N latitude, with an
Estimates of the total cultivated area and production of altitude of 2240 m above sea level in the Amhara
grass pea in Ethiopia was reported to be 83,522
National Regional State, Ethiopia. The soil is vertisol
hectares and 92,339 tonnes, respectively. Of this total
with pH 6.0. The weather variables during the testing
sum, northwest Ethiopia exceeds other regions, having
periods were generally conducive for normal growth
39,983 hectares (47.9 %) and 40,840 tonnes of
of grass pea.
production (44.2 %) (3).

The genotypes were planted in a randomized complete
Grass pea is a highly popular food and feed legume in
block design with two replications at a seeding rate of
the farming system due to tolerance of drought,
40 kg ha-1. A spacing of 60 cm between plots and 20
flooding and disease and its importance in
cm between rows were used, with a plot size of 0.8 m
ameliorating soil fertility (2). It is commonly grown as
x 4 m. The materials were evaluated for stand percent,
a double crop after the cereals tef or barley. Despite its
days to flowering, days to maturity, plant height (cm),
importance, the presence of the neurotoxin β-N-oxalyl
number of pods per plant, number of seeds per pod,
L-a, β-diaminopropanoic acid (ODAP) is a
100 seed weight (g) and grain yield (kg ha-1). ODAP
discouraging factor for grass pea production.
content analysis on seed was done at Addis Ababa
Irreversible crippling can occur, if the seeds are
University, Department of Chemistry, using Rao
consumed as a major part of the diet for an extended
Method for the 2001/02 cropping season only.
period (1). To date no single improved variety has been
Analysis of variance was computed for each season
developed and released in Ethiopia due to the
using an MSTAT computer program (7).
inconsistency of ODAP content of the promising grass

pea genotypes across environments.

The development of biotechnology and its application
Results and Discussion
in grass pea has resulted in somaclones with
The result of agronomic performance of the tested
neurotoxin ODAP content of less than 0.1% (100 mg
ODAP\100gm seed) in India (5). As a result the genotypes is presented in Table 1. Genotypes showed
significant difference (P<0.05) in days to maturity,
government of India released one of the somaclones,
Bio L212 (Ratan), for cultivation (8). Low ODAP lines 100 seed weight and grain yield. The grain yield level
are also available at the International Center for ranged from 3705 to 5233 kg ha-1. ILAT-LS-K-290
Agricultural Research in Dry land Areas (ICARDA) was the best yielder (5233 kg ha-1) and early in
(6). A study was carried out to test the agronomic maturity; whereas the local check was later to mature,
performances and the neurotoxin ODAP content of lower in 100 seed weight and grain yield (3705 kg ha-
those lines developed by somacloning technique at 1) than the other tested lines. ILAT-LS-K-289 and
ICARDA. ILAT-LS-K-444 were promising, both with 0.104 %
ODAP (Table 1). ILAT-LS-K-288 and ILAT-LS-K-
33 were also promising with low ODAP contents of
0.119 and 0.125 % seed. The local variety scored the
5
Lathyrus Lathyrism Newsletter 4 (2005)

highest ODAP content, 0.252 %. The results clearly will be included in the regional as well as the national
indicate that the somaclone technique is promising in breeding programme for testing under different
developing grass pea varieties with ODAP content environmental conditions and for possible release.
presumed at a safe level for human consumption (<
0.2 %) (4) in the future. Therefore, the promising lines

Table 1. Agronomic performance of grass pea somaclones at Adet, northwest Ethiopia (2000/01 and 2001/02).

Variety Stand Days to Days to Plant Pods/ Seeds/ 100 seed Seed Seed
% flowering maturity height plant pod weight (g) yield ODAP
(cm) (kg/ha) (%)*
ILAT –LS-K-290 81 64 142 115 71 3 7.3 5233 0.168
ILAT –LS-K-30 81 66 143 111 59 4 8.7 4317 0.216
ILAT –LS-K-104 83 66 145 121 69 4 10.2 5064 0.140
ILAT –LS-K-33 85 64 143 119 63 3 8.9 4261 0.125
ILAT –LS-K-299 84 67 147 112 56 3 7.2 4300 0.206
ILAT –LS-K-289 82 65 143 121 72 4 7.5 4807 0.104
ILAT –LS-K-444 82 64 144 114 64 4 7.6 4464 0.104
ILAT –LS-K-387 78 64 145 122 74 3 7.9 4405 0.211
ILAT –LS-K-190 78 64 142 112 64 4 7.6 4468 0.202
ILAT –LS-K-288 82 66 145 104 56 4 8.5 4209 0.119
ILAT –LS-K-390 85 57 143 104 56 4 7.4 4206 0.168
Local variety 86 54 149 126 67 4 6.5 3705 0.259
Mean 82 64 114 115 64 4 7.9 4462 -
S.E. 6.69 1.44 1.18 8.38 18.2 0.32 0.53 504 -
LSD (5%) 19.6 4.2 3.4 24.6 53.4 0.95 1.55 1478 -
CV (%) 11.5 3.2 1.2 10.3 40.3 12.9 9.4 16.0 -
*2001/02 only

References
1. Campbell CG. 1997. Grass pea (Lathyrus sativus 5. Mehta S.L. 1997. Plant biotechnology for
L.). Promoting the Conservation and use of removal of ODAP from Lathyrus. In: R.
underutilized and neglected crops. 18. Institute of Teklehaimanot and F. Lambein (eds) Lathyrus
Plant Genetic and Crop Plant Research, and Lathyrism: A Decade of Progress. University
Gatersleben/International Plant Genetic of Ghent, pp 103.
Resources Institutes, Rome, Italy. 6. Moneim, AM. Saxena MC, El-Saleh A, Nakkoul
2. Campbell CG, Tiwari KR. 1997. Breeding grass H. 1997. The status of breeding grass pea
pea for reduced seed levels of neurotoxin (Lathyrus sativus) for improved yield and quality
(ODAP). In: R. Teklehaimanot and F. Lambein at ICARDA. In: R. Teklehaimanot and F.
(eds) Lathyrus and Lathyrism: A Decade of Lambein (eds) Lathyrus and Lathyrism: A Decade
Progress. University of Ghent, pp. 85-86 of Progress. University of Ghent, pp. 81-82.
3. Dahiya BS. 1976. Seed morphology as an 7. MSTATC. 1989. A micro-computer statistical
indicator for low neurotoxin in Lathyrus sativus. program for experimental design, data
Qual Plant-Pl Fds Hum Nut 25, 391-394. management and data analysis. Michigan State
4. Central Statistics Authority, Ethiopia (CSA). University. Crop and Soil Science, Agricultural
2001/02. Agricultural Sample Survey. Addis Economics and Institute of International
Ababa, pp. 27. Agriculture, Michigan, USA.
8. Santha IM, Mehta SL. 2001. Development of low
ODAP somaclones of Lathyrus sativus. Lathyrus
Lathyrism Newsletter 2, 42-45.

6
Lathyrus Lathyrism Newsletter 4 (2005)

Search for resistance to crenate broomrape (Orobanche crenata) in Lathyrus

J.C. Sillero1*, J.I. Cubero2, M. Fernández-Aparicio3 and D. Rubiales3

1. CIFA, Dep. Mejora y Agronomía, Apdo. 3092, E-14080 Córdoba, Spain.
2. ETSIAM-UCO, Dep. Genética, Apdo. 3048, E-14080 Córdoba, Spain.
3. CSIC, Instituto de Agricultura Sostenible. Apdo.4084, E-14080 Córdoba, Spain.

* Email: [email protected]

Updated from original publication: Sillero JC, Cubero JI and Rubiales D. 2001.
Resistance to broomrape (Orobanche crenata) in Lathyrus.
7th International Parasitic Weed Symposium, (eds. A. Fer et al.), pp. 224-227,
Faculté des Sciences,Nantes, France.

Introduction Material and Methods
Grass pea has been widely cultivated in South Asia Fourteen accessions belonging to 10 different species of
and the Mediterranean area since antiquity for food the genus Lathyrus (Table 1) were screened for
and feed uses(4). Its cultivation was in continuous resistance to broomrape under field conditions in
recession in last decades, but there is an increasing 1995/96 in at Córdoba (Spain). Accessions were kindly
interest in its reintroduction in the cropping systems in provided by IPK (Germany) and USDA (USA). Each
marginal areas due to its adaptability to unfavourable accession was sown in a 1m row, surrounded by four
environments (4) and its beneficial effects on rows of a faba bean susceptible check (cv. Prothabon).
subsequent crops. Recently, plant breeders have been The sowing took place on 26 November 1995, in a field
working to improve its utilities (2) as well as to reduce heavily infested with O. crenata seeds. Hand weeding
anti-nutritional factors responsible for lathyrism (1). An was done when required, but no herbicides were applied.
additional priority for breeding L. sativus and L. The intensity of broomrape attack was evaluated at crop
cicera is resistance to crenate broomrape (Orobanche maturity (from early June), by counting the final number
crenata) (4). of emerged and non-emerged broomrape shoots per
plant. Data were expressed as a percentage of the mean
O. crenata is a holoparasitic weed that seriously of its four surrounding rows of Prothabon (=100%). The
attacks legume crops, such as faba beans, lentils, peas, most resistant lines were studied in a second field
chickpeas, grass peas and vetches (5,6,9). Different season, with field design and evaluation criteria
control methods have been proposed to avoid this similar to those described above.
serious problem, but none has been completely
effective and the development of resistant cultivars is The resistance found in two of the most resistant
a major need (8,9). Breeders are actively working on accessions was confirmed in pots and petri dishes
this matter and resistance to O. crenata has been experiments. The faba bean cv. Prothabon and a pea
found both in cultivated and wild legume species (7,9). variety (cv. Messire) were included as susceptible
The genus Lathyrus can be satisfactorily grown in checks. For the pot experiment five day old plants
marginal areas, so it is necessary to develop varieties were transplanted into 1 litre plastic pots filled with
resistant to broomrape to prevent this parasitic weed vermiculite, previously mixed with 25 mg of
problem in infested areas. broomrape seeds (about 8000 seeds). Each genotype
was represented by 12 plants, 1 plant/pot. When the
The purpose of this study was to search for sources of plants were mature, 90 to 120 days after sowing, the
resistance to O. crenata in different species of the plants were extracted, the roots were washed in water
genus Lathyrus. and the number of broomrape tubercles was counted.
The petri dish experiment was carried out using the
procedure described by Sauerborn (10). One month
after transplanting the seedlings, 500 seeds per dish
were studied and classified to determine the
percentages of germination and the number of
tubercles per plant.

7
Lathyrus Lathyrism Newsletter 4 (2005)

Table 1. Emerged and non-emerged broomrape attacks in selected accessions of Lathyrus, under field
conditions, during the growing seasons 1995/96 and 1996/97 in Córdoba (Spain)1.

1995/96 season 1996/97 season
Emerged Non-emerged Emerged Non-emerged
Accession Code Species shoots (%) shoots (%) shoots (%) shoots (%)
Lat-340 PI 255365 Lathyrus annuus 246 380
Lat-323 LAT 150/88 Lathyrus aphaca 73 91
Lat-341 PI 227529 Lathyrus aphaca 145 727
Lat-321 LAT 201/89 Lathyrus cicera 66 205
Lat-343 PI 208307 Lathyrus cicera 90 141
Lat-322 LAT 165/86 Lathyrus clymenum 0 0 0 0
Lat-344 PI 283488 Lathyrus clymenum 0 0 0 11
Lat-342 PI 229794 Lathyrus choranthus 0 123 16 34
Lat-345 PI 358859 Lathyrus gorgoni 34 129
Lat-348 PI 358864 Lathyrus inconspicuus 74 175
Lat-320 LAT 340/92 Lathyrus ochrus 0 0 0 0
Lat-352 PI 271361 Lathyrus ochrus 0 0 0 0
Lat-354 PI 165528 Lathyrus sativus 130 746
Lat-356 PI 269921 Lathyrus szowitsii 160 139
1 Emerged and non-emerged broomrapes referred to the susceptible faba bean check, cv. Prothabon (=100%).

Table 2. Established broomrape in pot and petri dishes and germination of broomrape seeds in petri dishes in
two selected accessions of Lathyrus and two susceptible checks.

Pot experiment Petri dishes experiment
No. of established % No. of tubercles/
Line Species broomrapes/ plant germination plant
Messire Pisum sativum 9.4 a 53.2 a 26.4 a
Protabon Vicia faba 7.6 a 37.6 b 13.3 b
Lat-320 Lathyrus ochrus 0.0 b 0.2 c 0.0 c
Lat-322 Lathyrus clymenum 0.0 b 0.1 c 0.0 c
Data with the same letter per column are not significantly different (P<0.05, Duncan test).

Results

In none of the accessions of L. clymenum and L. Results of in vitro experiments showed that no
ochrus studied was there any broomrape emergence broomrape was installed in any of the accessions, in
(Table 1), although in one accession of L. clymenum a neither pots nor in petri dishes (Table 2), which
few non-emerged tubercles were found in the second suggests a barrier to the broomrape establishment.
field season. Low broomrape emergence but high However, when the germination of the broomrape
levels of non-emerged tubercles were recorded in both seeds was recorded, almost no germination was found
seasons for the L. choranthus accession studied. All in any of the accessions, so the low germination of the
the accessions of the species L. annuus, L. aphaca, L. broomrape seeds seems to be the main barrier to the
cicera, L. gorgoni, L. inconspicuus, L. sativus and L. broomrape attack.
szowitsii were moderately to highly susceptible, all
with more than 34% emerged broomrape shoots.
8