Zentralbl. Mikrobiol. 146 (1991),137-141

Gustav Fischer Verlag lena

[Department of Microbiology, Haryana Agricultural University, Hisar, India]

Influence of Host, Moisture and Native Rhizobial Population on NoduleOccupancy in Chickpea (Cicer arietinum)

A. L. KHURANA, P. K. SHARMA and S. S. DUDEJA

Key words: Cicer arietinum, Rhizobium, nodule occupancy

Summary

Seven chickpea (Cicer arietinum L.) cultivars belonging to three nodulation groups low nodulating cultivarsL 550, H208, moderate nodulating cultivars, BG 209, Pant G 114 and C 235 and good nodulating cultivars K 850and H 75-35 were used in the prestent study. Nodule occupancy by an inoculant Cicer Rhizobium strain CM-I variedunder two sets of conditions. Moderate nodulating cultivars showed maximum acceptance of the Cicer Rhizobiumstrain CM- I, high nodulating and low nodulating cultivars were at par. The strain acceptability was primarilydependent on host and secondary role was played by moisture and native rhizobial population.

Zusammenfassung

Sieben Varietaten der Kichererbse (Cicer arietinum L.) konnten 3 Kniillchenbildungs-Gruppen zugeordnetwerden: Varietat L 550 und H 208 mit geringer, Varietat BG 209, Pant G 114 und C 235 mit mittIerer undVarietiiten K 850 und H 75-35 mit guter Kniillchenausbildung. Die Besetzung der Kn6llchen mit einem inokuliertenRhizobium-Stamm CM-I variierte entsprechend der Untersuchungsbedingungen.

Die Varietaten mit mittlerer Kniillchenbildung zeigten eine maximale Besetzung des Cicer Rhizobium-StammesCM-I und die mit geringer bzw. hoher Kniillchenbildung eine mittlere Annahme. Ober die Etablierung einesinokulierten Rhizobium-Stammes entscheidet in crster Linie die Wirtspflanze, aber auch Feuchtigkeit und nativeRhizobium-Population spielen dabei eine Rolle.

One of the most important and pressing problem in the ecology of Rhizobium is its ability toform nodules because certain native rhizobia are more competitive than the inoculant strains fornodule formation. A particular strain may be highly competetive due to many factors, some ofwhich are not yet fully understood. The attributes of possible relevance to the success of a strainincludes: strain, host, strain host interactions and environment. Highly effective strains variedgreatly in their degree of competetiveness (GAUR and LOWTHER 1982a; 1982b; MAY andBOHLOOL 1983; MOAWAD et a1. 1984; RAFIQUE UDDIN et a1. 1984). Success of an inoculant strainis affected by the preferential recognision of the strain by the host (CALDWELL and VEST 1968;MASTERSON and SHERWOOD 1974; JONES and HARDARSON 1979) and by soil type (MOAWAD andBOHLOOL 1984). SCHMIDT and ROBERT (1985) showed that the interactions which occur in therhizosphere during very early plant growth are critical in determining the outcome of competitionamong R. japonicum strains. Other factors affecting the competitiveness of a strain are pH(DOUGHRI and BOTTOMLEY 1984), root temperature (ROUGHLEY et a1. 1980) and environmentalconditions (DUDEJA and KHURANA 1988).

In view of this an experiment was conducted at two locations with different native rhizobialpopulations using 7 chickpea cultivars (Cicer arietinum L.) to ascertain whether it is nativerhizobial population or host selection or soil condition which determines the competitive ability ofan inoculant strain CM- 1 under field conditions.

138 A. L. KHURANA et al.

Materials and Methods

CultivarsSix cultivars of desi gram (H 75-35, BG 209, H 208, Pant G 114, K 850, C 235) and one kabuli cultivar L 550

were selected for present investigation.

Rhizobium strain

Cicer-Rhizobium strain CM-I resistant to streptomycin (200 Ilg . ml- 1) found efficient under field conditions

was used in competition study. The strain was procured from International Research Institute for Semi-Arid Tropics(ICRISAT) Patancheru, India.

Field experiment

Field experiment was carried out at Haryana Agricultural University Research farm, 200 m above sea level atlatitude of 29° 10' N and longitude 75° 46' E at two different fields with soil characteristics given in Table I. In one

Table. I. Characteristics of the two fields where experiments was conducted.

Characteristics

Soil typeSoil pH (H20 I: 2)Organic CElectrical conductivity (H20 I :2)Total NP (Olsen)Moisture

Field (FI)

Sandy loam (Less sand)7.60.42%0.29dsm- 1

0.065%10 ppmPresown irrigation(12-13%)

Field (F2)

Sandy loam (More sand)8.00.31 %0.18dsm- 1

0.007%7 ppmConserved moisture(6-7%)

field (F1), presown irrigation was given whereas in second field (F2), crop was sown on conserved moisture. Totalrainfall during the experiment was 13 mm. The native chickpea rhizobial population was 5.4 x 105 and 2.3 X 103

rhizobia g-I of soil in F1 and F2 fields respectively. The experiment was conducted in triplicate.

Competitiveness

Competitive ability of strain CM-l under two fields F1 and F2 was determined by antibiotic resistance marker(200 Ilg . ml- I of streptomycin). After 45 d of planting, 12 plants from 6 random locations from each replicate wereuprooted. All the nodules were detached and pooled. At least 50 nodules were tested for the presence or absence ofinoculant strain by streaking the nodule sap on yeast extract mannitol agar plates containing 200 Ilg' ml- 1 ofstreptomycin or without streptomycin as control plates. Plates were incubated at 28 ± 2 °C for 7 d. The streaksshowing Rhizobium like colonies were taken as positive, indicating thereby that nodules were formed by theinoculant strain.

Other observations like nodule number, nodule dry weight were also recorded at 45 d sampling. Grain yield perplot was recorded after harvesting the crop at maturity.

Results

Depending upon the number of nodules formed by native rhizobia, the seven chickpeacultivars used in the present investigation were grouped into three categories. First group showingnodules upto 10 nodules per plant (poor nodulation) included cultivars L 550 and H 208. Secondgroup included three cultivars BG 209, Pant G 114 and C 235, these showed 11 - 20 nodules perplant (moderate nodulation). Third group included two high nodulating (40 nodules per plant)cultivars K 850 and H 75-35.

In field F j Cicer Rhizobium strain CM-l formed nodules to a maximum of 28 % in case of cvPant G 114 (Table 2). Nodule occupancy decreased from 28 to 6 % in cv K 850. However in field

Nodule Occupancy in Chickpea 139

Table. 2. Nodule occupancy of strain CM-I in chickpea cultivars under two field condition.

Cultivars % nodule formed by strain CM-I

Field (Fl) Field (F2)

Low NodulatingL550 10.7 2.5H208 10.9 5.8

Moderate NodulatingBG209 20.7 18.6Pant 114 28.0 15.6C235 15.7 10.7

High NodulatingK850 6.0 8.7H75-35 8.7 7.5

Table. 3. Overall nodule occupany by strain CM-I in different chickpea cultivars.

Field % nodule formed by strain CM-l

Low nodulating Moderate nodulating Good nodulating Mean

Field Fl 10.8 21.5 7.4 14.4Field F2 4.2 15.0 8.1 10.0Mean 7.5 18.3 7.8

Table. 4. Grain yield of different chickpea cultivars under two sets of condition.

Variety

H75-35BG 209Pant G-114L550K850-3/27H 208C 235

C.D. at 5%

Grain yield (q/ha)

Field (FI) Field (FI)

Control Inoculated % increase Control Inoculated % increase

24.30 24.56 1.06 11.92 13.32 11.7431.30 33.19 5.93 14.42 18.59 28.9135.00 37.44 6.97 17.49 18.49 6.2826.27 29.83 13.55 9.71 7.7526.71 23.95 10.27 12.21 18.2837.78 38.10 1.08 13.32 13.32 028.40 33.70 19.7 12.00 14.00 16.2

Fl (field) F2 (field)

Inoculation NS 1.92Cultivars 6.35 3.21Inoculation x cultivars NS 2.15

F2 the maximum competitiveness of 18.6 % was recorded with cv BG 209 followed by Pant G 114and least by cv L 550. The acceptance of strain CM-l by all the seven chickpea cultivars wasvariable and in general the competitiveness of strain CM-l was more in field F1 as compared tofield F2 . Moderate nodulating cultivars BG 209, Pant G 114 and C 235 formed maximum numberof nodules by CM-l to an extent of 21.5 and 15 % under F1 and F2 field conditions respectivelyand followed by low nodulating cultivars L 550 and H 208 (Table 3). The good nodulating

140 A. L. KHURANA et aI.

cultivars K 850 and H 75-35 showed comparatively less number of nodule formation by theinoculant strain. On overall mean basis low and high nodulating cultivars behaved similarly.Competitiveness of CM-l on overall mean basis was 14.4% in field F1, 10.0% in field F2.

Inoculation with CM-l strain increased grain yield significantly under F2 field conditions andincrease was non-significant under F1 field conditions (Table 4). The increase in grain yield indifferent cultivars ranged between 1.0-18.7 and 6.3-28.9 % under two sets of conditions.Maximum grain yield was recorded in cv H 208 (38.1 q . ha-1) in field F1 and in cv Pant G 114and BG 209 in field F2• No correlation between competitiveness and increase in grain yield wasobserved in any of the cultivars under both conditions.

Discussion

The results indicates specific host effect in determining the relative success in nodulation byrhizobial strain. All the seven chickpea cultivars differed considerably in their selection for theinoculated strain under two sets of conditions.

Such variations has also been reported elsewhere (CALDWELL and VEST 1968; MASTERSONand SHERWOOD 1974; JONES and HARDARSON 1979). On the basis of strain acceptance it may beinferred that high nodulating cultivars exerted least selective pressure for strain acceptance fromnative population. Whereas moderate nodulating cultivars exerted maximum selective pressurefor strain acceptance. High nodulating cultivars formed maximum number of nodules but becauseof least selective pressure it could accept the inoculated strain to a lesser extent than that ofmoderately nodulating cultivars. Likewise low nodulating cultivars being very specific for strainacceptance, formed low number of nodules and could accept inoculated strain to a lesser extentthan that of moderate nodulating cultivars.

Nodule occupancy by strain CM-l decreased under field F2 as compared to field F1conditions. This was probably due to formation of less number of infection thread due to lowmoisture as reported by WORRAL and ROUGHLEY 1976 and thereby leading to the formation oflesser number of nodules. Although native population was high under field F1 conditions than thatof field F2 conditions yet inoculated strain formed more nodules under field F1 conditionsindicating that soil moisture may be one of the limiting factor for nodule occupancy. Similar strainacceptance under field F I and F2 by different host showed that host is primarily controlling thenodule occupancy and native rhizobial population and moisture are the secondary factors. Thepresent study suggest that compatible genotypes and Rhizobium strain must be screened underdifferent sets of conditions.

References

CALDWELL, B. E., VEST, G.: Nodulation interactions between soybean (Glycine max.) genotypes and serogroups ofRhizobium japonicum. Crop Sci. 8 (1968), 680-682.

DUDEJA, S. S., KHURANA, A. L.: Survival and competitiveness of Bradyrhizobium sp. in the rhizosphere ofpigeonpea (Cajanus cajan). BioI. Fertil. Soils 7 (1988), 63-67.

DUGHARI, M. H., BOTTOMLEY, P. J.: Soil acidity and the competition of an indigenous population of Rhizobiumtrifolii in nodules of different cultivars of Trifolium subterraneum L. Soil BioI. Biochem. 16 (1984), 405-411.

GAUR, Y. D., LOWTHER, W. L.: Competitiveness and persistence of introduced rhizobia on over sown clover:influence of strain, inoculation rate and lime pelleting. Soil BioI. Biochem. 14 (l982a), 99-102.

GAUR, Y. D., LOWTHER, W. L.: Competitiveness and persistence of strains of Rhizobium trifolii in relation toinoculation level and lime peUeting on white clover sown into cultivated soil. N. J. Agric Res. 25 (l982a),277-280.

JONES, D. G., HARDARSON, G.: Variation within and between white clover varieties in their preference for strains ofRhizobium trifotii. Ann. Appl. BioI. 92 (1979),221-228.

MASTERSON, C. L., SHERWOOD, M. T.: Selection of Rhizobium trifolii strains by white and subteranean clovers. Ir.J. Agric Res. 20 (1974), 827-841.

MAY, S. N., BOHLOOL, B. B.: Competition among Rhizobium leguminosarum strains for nodulation of lentils (Lensesculenta). Appl. Environ. Microbiol. 44 (1983), 960-965.

Nodule Occupancy in Chickpea 141

MOAWAD, H. A., BOHLOOL, B. B.: Competition among Rhizobium spp. for nodulation of Leucaena leucocephalain two tropical soils App!. Environment Microbio!. 48 (1984), 5-9.

MOAWAD, H. A., ELLIS, W. R., and SCHMIDT, E. L.: Rhizosphere response as factor in competition among threeserogroups of indigenous Rhizobium japonicum for nodulation of field grown soybeans. App!. Environ.Microbio!. 47 (1984), 607-612.

RAFIQUE UDDIN, M., LAUGHLIN, W. M. C., AHMED, M. H.: Competition between inoculum and native rhizobiafor nodulation of cowpea (Vigna unquiculata L. Walp): Use of a dark nodule strain. Plant. Soil 81 (1984),305-307.

ROUGHLEY, R. J., BROMFIELD, E. S. P., PULVER, E. L., and DAY, J. M.: Competition between species ofRhizobium for nodulation of Glycine max. Soil BioI. Biochem. 12 (1980), 467-470.

SCHMIDT, E. L., and ROBERT, F. M.: Recent advances in the ecology of Rhizobium. In: Nitrogen Fixation ResearchProgress (Eds.: EVANS, H. J., BOTTOMLEY, P. J., NEWTON, W. E.) Martinus Nijhoff Publishers 1985,379-385.

WORALL, V. S., ROUGHLEY, R. J.: The effect of moisture stress on infection of Trifolium subterraneum L. byRhizobium trifolii. Dang. J. Exp. Bot. 27 (1976). 1233-1241.

Author's address: Dr. S. S. DUDEJA, A. L. KHURANA, P. K. SHARMA, Dep. of Microbiology, Haryana AgriculturalUniversity, Hisar 125004, India.