Symbiotic effectiveness of streptomycin-resistant Rhizobium spp. mutants of chickpea (Cicer arietinum L.)

Summary Antibiotic-resistant Rhizobium spp. strains have been used in ecological studies of legumerhizobia symbiosis. It has been suggested that in the course of acquiring resistance against high doses of antibiotics, rhizobia might lose their symbiotic effectiveness. Evidence both for and against this argument has been presented (Kremer and Peterson 1982; Materon and Hagedron 1983). This communication reports our experience with streptomycin-resistant (Str⁺) mutants of chickpea Rhizobium spp. strains. Parent strains were used as controls.Research paper No. 5233 from the Experiment Station, G.B.P.U.A.&T. Pantnagar, Nainital     

Differential responses of chickpea (Cicer arietinum L.) — Rhizobium combinations to saline soil conditions

Summary Chickpea cultivars (Cicer arietinum L.) and their symbiosis with specific strains of Rhizobium spp. were examined under salt stress. The growth of rhizobia declined with NaCl concentrations increasing from 0.01 to 2% (w : v). Two Rhizobium spp. strains (F-75 and KG 31) tolerated 1.5% NaCl. Of the 10 chickpea cultivars examined, only three (Pusa 312, Pusa 212, and Pusa 240) germinated at 1.5% NaCl. The chickpea — Rhizobium spp. symbiosis was examined in the field, with soil varying in salinity from electrical conductivity (EC) 4.5 to EC 5.2 dSm^-1, to identify combinations giving satisfactory yields. Significant interactions between strains and cultivars caused differential yields of nodules, dry matter, and grain. Four chickpea — Rhizobium spp. combinations, Pusa 240 and F-75 (660 kg ha^-1), Pusa 240 and IC 76 (440 kg ha^-1), Pusa 240 and KG 31 (390 kg ha^-1), and Pusa 312 and KG 31 (380 kg ha^-1), produced significantly higher grain yields in saline soil.     

Effect of inoculum rate on the performance of chickpea (Cicer arietinum L.) Rhizobium strains in the field

Summary The influence of three inoculum rates on the performance of three chickpea (Cicer arietinum L.) Rhizobium strains was examined in the field on a Mollisol soil. Increasing amounts of inoculum improved the performance of the strains. A normal dose (10⁴ cells per seed) applied at different intervals gave non-significant increases in nodulation, nitrogenase activity (acetylene reduction assay), nitrogen uptake and grain yield. A ten-fold increase in inoculum increased nodule number, shoot dry weight, nitrogenase activity (ARA) and grain yield, but increases over the control were significant only for nodule dry weight and nitrogen uptake by shoot and grain. The highest level of inoculum (100 × normal) significantly increased nodule dry weight, grain yield, total nitrogenase activity (ARA) and nitrogen uptake by shoot and grain. Strain TAL 620 was more effective than the other two. Combined nitrogen (60 kg N ha^–1) suppressed nodulation and nitrogenase activity (ARA).Research paper No. 4345 from the Experiment Station, G. B. P. U. A. & T., Pantnagar, Nainital, U. P.     

Displacement of native rhizobia nodulating chickpea (Cicer arietinum L.) by an inoculant strain through soil solarization

Summary Soil solarization greatly reduced the native chickpea Rhizobium population. With inoculation, it was possible to increase the population of the Rhizobium in solarized plots. In the 1st year, 47% nodulation was obtained with chickpea inoculant strain IC 59 when introduced with a cereal crop 2 weeks after the soil solarization and having a native Rhizobium count of <10 g^-1 soil, and only 13% when introduced 16 weeks after solarization at the time the chickpeas were sown, with 2.0×10² native rhizobia g^-1 soil. In the non-solarized plots inoculated with 5.6×10³ native rhizobia g^-1 soil, only 6% nodulation was obtained with the inoculant. In the succeeding year, non-inoculated chickpea was grown on the same plots without any solarization or Rhizobium inoculation. The treatment that showed good establishment of the inoculant strain in year 1 formed 68% inoculant nodules. Other treatments indicated a further reduction in inoculant success, from 1%–13% to 1%–9%. Soil solarization thus allowed an inoculant strain to successfully displace the high native population in the field and can serve as a research tool to compare strains in the field, irrespective of competitive ability. In year 1, Rhizobium inoculation of chickpea gave increased nodulation and increased plant growth 20 and 51 days after sowing, and increased dry matter, grain yield, and grain protein yield at maturity. These beneficial effects of inoculation on plant growth and yield were not measured in the 2nd year.Submitted as Journal Article No. JA 945 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, Andhra Pradesh 502 324, India     

Evaluation of 15N-isotope dilution for measurement of nitrogen fixation in chickpea (Cicer arietinum L.)

Summary N accumulation, nodulation, and acetylene reduction activity were measured at frequent intervals during the growth of two chickpea genotypes, and N₂ fixation was estimated by an isotope-dilution method, using safflower as a non-N₂-fixing reference. Safflower was more efficient at N uptake than both the chickpea genotypes for at least the first 50 days and thus could not be used as an accurate reference control. We recommend that further work should employ non-nodulatiog genotypes of chickpea as reference plants and use slow-release forms of 15N fertilizer. Direct genotype comparison by isotope dilution estimated that genotype K 850 fixed 16–18 kg ha^–1 more N than G 130, and this difference was supported by the greater nodule mass and acetylene reduction activity in the K 850 cultivar. Inoculation with an ineffective chickpea Rhizobium sp. led to 69% nodulation on cultivar K 850 but only 33% on G 130. While nodule weight, N uptake, and acetylene reduction activity decreased with inoculation in K 850, the isotope dilutions were similar for both inoculation treatments. The lack of a significant effect on N₂ fixation was ascribed to the partial success of inoculant establishment.Published as Journal Article No. JA 692 of the International Crops Research Institute for the Semi Arid Tropics, Patancheru, A.P. 502324, India     

Influence of irrigation on the distribution and control of the nematode Meloidogyne javanica by the biocontrol bacterium Pasteuria penetrans in the field

Pasteuria penetrans, a bacterial parasite of plant-parasitic nematodes, is used to control root-knot nematode Meloidogyne spp. populations in vegetable crops. But its efficiency is variable, mostly because of the patchy distribution of the bacteria in arable fields. As the infective P. penetrans are non-motile bacteria in soil, abiotic soil factors can affect the bacteria–nematode relationships. An epidemiological study, conducted in a vegetable field, showed that abiotic factors such as irrigation, soil water holding capacity and texture, affected the efficiency of P. penetrans. A correspondence analysis between these abiotic factors and the density of P. penetrans spores in the soil, and the proportion of Meloidogyne javanica juveniles infected by the bacteria, revealed that irrigation affected directly the distribution of the spores in soil pores related to their passive transport by water flow. Laboratory experiments conducted on the passive transport of spores confirmed that intensive irrigation leached the spores down the soil profile and decreased the percentage of infected Meloidogyne juveniles.     

Comparison of some endogenous hormone levels in different parts of chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.)

The objective of this study was to detect plant hormone levels in different plant parts of chickpea, and to compare cultivars with different characteristics such as simple leaf vs. normal leaves, simple leaf vs. bipinnate leaf, kabuli type vs. desi types, double-podded vs. single-podded. It was found that there was a great variation in basic plant hormone levels among genotypes, and plant hormone concentrations varied according to leaf type, pod and seed characteristics in these genotypes. IAA level in leaves was the highest in ICC 6119 (bipinnate leaf) and followed by ICC 552 (double-pod) and Kusmen 99 (simple leaf), while it was highest level in pods of ICC 552 (double-pod). Zeatin concentration in leaves and pods was the highest level in ICC 6119 (bipinnate leaf). GA₃ concentration was the highest in leaves of ICC 6119 (bipinnate leaf). ICC 552 (double-pod) has the highest GA₃ in leaves. It was postulated that leaf type and pod characteristics may be related to hormones induced growth and development.     

Clustering of Chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) Accessions

The variation in 20 morphological and agronomical traits has been evaluated in a set of chickpea genetic resources from four countries. Data indicated differences between accessions in leaf, flower, pod, and seed traits and characteristics, as well as in vegetation period. Multivariate analyses of these data segregated chickpeas into groups. Polymorphism in seed glutelines was absent, while variation in seed prolamines was very limited. DNA amplification patterns have been analyzed by semi-arbitrary primers and by specific microsatellite primers. Only some of semi-arbitrary primers generated usable DNA banding patterns, moreover interpretation of these patterns can be more or less difficult. On the contrary specific primers amplifying microsatellites at the specific loci generated unambiguous and reproducible differences between chickpeas.     

The extent of embryo and endosperm growth following interspecific hybridization between Cicer arietinum L. and related annual wild species

In vivo interspecific pollinations were performed and immature seed development investigated by histological methods in order to study crossability barrier(s) in Cicer L. species wide hybridization. Seven of the eight wild annual Cicer species, belonging to the secondary and tertiary gene pools, were used in reciprocal crosses with the cultivated chickpea. It was confirmed that the zygote was formed in all interspecific crosses. The embryos showed continued and retarded growth at different rate in various crosses, but eventually aborted at an early pro-embryo stage in all crosses except C. arietinum L. ×C. echinospermum Dav. Reciprocal cross differences were observed in early embryo growth rate and could have implications in obtaining hybrids. This study further emphasizes the necessity for developing appropriate and efficient in vitro procedures for rescuing immature globular hybrid pro-embryos, which will make the wild Cicer gene resources amenable to chickpea improvement.     

RAPD and ISSR fingerprinting in cultivated chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) and its wild progenitor <Emphasis Type="Italic">Cicer reticulatum</Emphasis> Ladizinsky

Detection of genetic relationships between 19 chickpea cultivars and five accessions of its wild progenitor Cicer reticulatum Ladizinsky were investigated by using RAPD and ISSR markers. On an average, six bands per primer were observed in RAPD analysis and 11 bands per primer in ISSR analysis. In RAPD, the wild accessions shared 77.8% polymorphic bands with chickpea cultivars, whereas they shared 79.6% polymorphic bands in ISSR analysis. In RAPD analysis 51.7% and 50.5% polymorphic bands were observed among wild accessions and chickpea cultivars, respectively. Similarly, 65.63% and 56.25% polymorphic bands were found in ISSR analysis. The dendrogram developed by pooling the data of RAPD and ISSR analysis revealed that the wild accessions and the ICCV lines showed similar pattern with the dendrogram of RAPD analysis. The ISSR analysis clearly indicated that even with six polymorphic primers, reliable estimation of genetic diversity could be obtained, while nearly 30 primers are required for RAPD. Moreover, RAPD can cause genotyping errors due to competition in the amplification of all RAPD fragments. The markers generated by ISSR and RAPD assays can provide practical information for the management of genetic resources. For the selection of good parental material in breeding programs the genetic data produced through ISSR can be used to correlate with the relationship measures based on pedigree data and morphological traits to minimize the individual inaccuracies in chickpea.     

Performance of chickpea (Cicer arietinum L.) to bio-fertilizer and nitrogen application in arid condition

Abstract

Field experiments were conducted during 2013–2014 at Tashkent, Uzbekistan to evaluate the performance of chickpea variety “Jakhongir” with the variable proportion of nitrogen (N) and bio-fertilizer inoculation in the moderate saline (5.6?±?0.6?dSm^?1) soil condition. The studied treatments were No control (non-fertilized), N1 mineral-N (50?kg?N?ha^?1), N2, mineral-N (75?kg?N?ha^?1), N3, mineral-N (100?kg?N?ha^?1) equivalent 0%, 50%, 75%, and 100% from recommended rate for chickpea, Rhizobium inoculation (Bio)?+?No control, Rhizobium inoculation (Bio)?+?N1, Rhizobium inoculation (Bio)?+?N2, and Rhizobium inoculation (Bio)?+?N3. Seed inoculation with Rhizobium was significantly superior over no inoculation treatments at all rate of N fertilization. The middle rate of N fertilization 75?kg?N?ha^?1 combined with biofertilizer inoculation had of superior effect on chickpea, producing 73.2% more yield (1.68?Mg ha^?1), oil, protein, and sugar content performed 16.4%; 15.0%, and 17.9% higher value, respectively, in comparison to control.     

Effect of irrigation and biofertilizer on water use,nodulation, growth and yield of chickpea (Cicer arietinum L.)

The field experiment studied the effect of irrigation [irrigation 15 days before sowing (DBS), irrigation 15 DBS + rice straw mulch, irrigation 7 DBS and irrigation 7 DBS + one irrigation at flower initiation] and biofertilizers [no biofertilizer (control), Rhizobium inoculation and Rhizobium inoculation + phosphate-solubilizing bacteria (PSB)] on chickpea growth. In mulch treatment, paddy straw mulch was applied at 4 t ha^?1 one day after pre-sowing irrigation and was retained until sowing. Pre-sowing irrigation at 15 DBS showed 28.7 and 30.0% less plant stand than irrigation applied 15 DBS + straw mulch and irrigation applied 7 DBS, respectively. Nodulation was significantly higher with irrigation 15 DBS + mulch and irrigation 7 DBS than with irrigation applied 15 DBS. The grain yield was 16.6, 20.3 and 44.0% higher in irrigation 15 DBS + mulch, irrigation 7 DBS and irrigation 7 DBS + irrigation at flowering treatments, respectively, over irrigation at 15 DBS. Rhizobium inoculation significantly improved the number of nodules and nodule dry weight compared with no treatment. Grain yield was significantly higher with Rhizobium than in untreated plots. Water use efficiency was highest when irrigation was applied 7 DBS.     

New sources of resistance to race Ro1 of the Golden nematode (Globodera rostochiensis Woll.) among reputed duplicate germplasm accessions of Solanum tuberosum L. subsp. andigena (Juz. et Buk.) Hawkes in the VIR (Russian) and US Potato Genebanks

Cultivated Solanum tuberosum L. subsp. andigena is well known as a rich source of valuable traits for potato breeding, especially for resistance to diseases and pests. The golden potato cyst nematode, Globodera rostochiensis Woll., is considered to be one of todays most serious hindrances to potato production in Europe and North America. Thus, the breeding of new cultivars that have resistance to PCN is of great importance. The USPG (USA) and VIR (Russian) potato genebanks, as well as others, maintain many samples of primitive cultivated and wild potato species originating from Latin America. Many of these samples are assumed to be genetically duplicated because the material in both genebanks came from the same original source. A joint investigation of new genotypes of subsp. andigena forms resistant to potato cyst nematode (PCN) was carried out on samples of subsp. andigena at VIR with reputed duplicate samples at USPG. After careful screening, 14 samples which possessed resistance to PCN were identified. A high level of this resistance was transmitted to sexual progeny at a high frequency for all of the selections. Eleven of the accessions found to be resistant have reputed duplicates in USPG that were not previously known to be resistant. Thus, this research not only broadens the choice of parents available for resistance breeding, but identifies model materials for future research to test the parity of PCN resistance among reputed duplicate samples in the two genebanks.     

Assessment of genetic diversity, and phylogenetic relationships based on ribosomal DNA repeat unit length variation and Internal Transcribed Spacer (ITS) sequences in chickpea (Cicer arietinum) cultivars and its wild species

Repeat unit length variation and internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA were used to assess genetic diversity, and phylogenetic relationships in chickpea (C. arietinum) cultivars, and its related wild species. Total genomic DNAs of 76 accessions of 10 Cicer species, belonging to three sections of the genus, were restricted with seven enzymes and the restriction fragments were hybridized to heterologous ribosomal clones of wheat pTa71 and Vicia faba probes Ver 6-5 and Ver18-6. A single repeat unit length class of 11.4 kb or 10.5 kb was recognized across Cicer accessions with pTa71. The intraspecific variation was negligible in those species where more than one accession was studied, except the four C. judaicum accessions, which were different from the rest. EcoRI and DraI digests gave two and one-two fragments, respectively. All the accessions produced three and three-five bands with BamHI and SacI, respectively. Both the accessions of C. yamashitae differed in their rDNA repeat unit length as well as restriction site variation. Maximum likelihood tree with rDNA RFLP recognized five clades which were more or less congruent with the previous data. Length of ITS-1 region was more variable (235–239 bp) than the ITS-2 region (212–213 bp). Cladistic analysis of ITS data revealed two major clades, clade I consisting of C. arietinum, C. reticulatum and C. echinospermum, and clade II comprised of C. judaicum, C. chorassanicum, C. bijugum and C. cuneatum. C. microphyllum grouped with the above four species. C. pinnatifidum was present as a separate branch. C. yamashitae emerged as the most distinct species.     

Pre-inoculation with arbuscular mycorrhizal fungi suppresses root knot nematode (<Emphasis Type="Italic">Meloidogyne incognita</Emphasis>) on cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>)

A pot experiment was conducted to evaluate the influence of pre-inoculation of cucumber plants with each of the three arbuscular mycorrhizal (AM) fungi Glomus intraradices, Glomus mosseae, and Glomus versiforme on reproduction of the root knot nematode Meloidogyne incognita. All three AM fungi tested significantly reduced the root galling index, which is the percentage of total roots forming galls. Numbers of galls per root system were significantly reduced only in the G. intraradices + M. incognita treatment. The number of eggs per root system was significantly decreased by AM fungus inoculation, no significant difference among the three AM fungal isolates. AM inoculation substantially decreased the number of females, the number of eggs g⁻¹ root and of the number of eggs per egg mass. The number of egg masses g⁻¹ root was greatly reduced by inoculation with G. mosseae or G. versiforme. By considering plant growth, nutrient uptake, and the suppression of M. incognita together, G. mosseae and G. versiforme were more effective than G. intraradices.     

Growth,nutrient uptake and yield parameters of chickpea (Cicer arietinum L.) enhance by Rhizobium and Azotobacter inoculations in saline soil

Abstract

A field experiment was conducted during two consecutive growing seasons (2013 and 2014) to evaluate the effects of inoculations with Rhizobium and Azotobacter on the growth and yield of two chickpea (Cicer arietinum L.) varieties under saline (5.8 dS m^?1) arid condition. The single treatment of either Rhizobium or Azotobacter exhibited to promote the growth of chickpea to some level, however, co-inoculation produced more effects and increased the shoot dry weight (30.3 and 26.4%), root dry weight (17.5 and 26.3%), nodule number (79.1 and 43.8 piece per plant), nitrogen content in roots (9.62 and 10.9%), in shoots (12.6 and 8.3%) and seed protein (7.1 and 4.3%) in both Flip06-102 and Uzbekistan-32 chickpea varieties compared to the control. Our studies showed that the highest yield response of 429 (27.9%) and 538 (23.9%) kg?ha^?1 over the control was revealed by the co-inoculation with Rhizobium and Azotobacter inoculants in Flip 06-102 and Uzbekistan-32, respectively. A new introduced Flip 06-102 chickpea variety was more salt tolerant and had higher root nodulation than the local Uzbekistan-32 chickpea variety. Nitrogen (N), phosphorus (P), and potassium (K) contents in the shoots and roots were significantly (p?Rhizobium plus Azotobacter could be applied to improve the vegetative growth and yield of chickpea and to alleviate the effects of salt stress.     

Synergistic effect of bio-inoculants on yield,nodulation and nutrient uptake of chickpea (Cicer arietinum L) under rainfed conditions

The use of efficient bio-inoculants in chickpea is the best way to increase crop productivity under rainfed conditions. To assess the combined effect of bio-inoculants on crop yield, field experiments were conducted during Rabi seasons at Research Station, Punjab Agricultural University, Ballowal Saunkhri, Punjab, India. The application of different bio-inoculants significantly improved number of pods, grain and straw yield of chickpea over the un-inoculated treatment. The combined application of Rhizobium + PSB?+?AM fungi?+?azotobactor inoculums as seed treatment with 75% of recommended phosphorus produced highest grain yield. The nodule count, nodule weight, per cent root colonization of AM fungi and different enzymes activities in soil were also highest in combined bio-inoculants treatment. The present study concluded that combined application of bio-inoculants (Rhizobium, PSB, AM fungi and azotobactor) can save 25% of recommended phosphorus by sustaining the crop yield and improving the soil health.     

Sampling strategies and screening of chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) germplasm for salt tolerance

Six hundred accessions of chickpea (Cicer arietinum L.) landraces and its wild relatives from 28 different countries, available at Australian Temperate and Field Crops Collection (ATFCC) were screened for tolerance to salt under greenhouse conditions using three sampling strategies; (1) random sampling of 200 accessions from different countries, (2) restricted random sampling of 200 accessions from geographical regions with salinity problems and high diversity (Middle East and West & South Asia) and (3) as for strategy 1 but with a reduced representation of accessions from the geographical regions used in strategy 2. Degree of salt tolerance was based on necrosis scores and shoot biomass reduction relative to unstressed controls at harvest after subjecting stressed plants to salt treatment from 21 to 42 days after sowing. There was a wide variation in salinity tolerance determined by both measures. For sampling strategies 1, 2 and 3 respectively; 24, 28 and 14% of accessions were salt tolerant. Accessions from the middle east and south Asian (regions with salinity problem, a long history of chickpea cultivation and high diversity) gave a higher probability (P < 0.01) of getting salt tolerant accessions.     

Effect of water flow on the mobility of the root-knot nematode Meloidogyne incognita in columns filled with glass beads, sand or andisol

In the present study, the migration of nematodes was studied in columns filled with three materials of different textures and chemical properties. The role of soil pores that enable root-knot nematode (Meloidogyne incognita) second stage juveniles (J2) to escape rapid water flow in soil was demonstrated using columns filled with glass beads, sand or andisol that maintained a constant water flow. Under a constant flow flux of 36 cm h⁻¹, living J2, dead J2 or anion bromine tracer (Br⁻) was injected in the middle of the column and then drainage water equivalent to two pore volumes (PV) was collected. The passive transport of the anion tracer in water flow could be explained by a convection dispersion equation. The dead J2 showed a pattern similar to that of Br⁻. However, the living J2 resisted movement in the water flow and remained in the column even at the highest water flow rate of 93.3 cm h⁻¹ in glass beads. The mobility of living J2 was affected by the filling materials; the number of J2 passing through the column was much lower in the andisol-filled column than in the other two columns but the total number of J2 in drainage water was 5% or less of the number injected for all columns. We suggest that J2 were affected not only by soil water flow but also by soil pore structure and have the ability of withstanding or avoiding movement in soil water flow.     

Growth,yield and water use efficiency of chickpea (Cicer arietinum): response to biochar and phosphorus fertilizer application

Field experiments were conducted during summer (2013/2014) and winter (2014) in two different soil types to evaluate the effect of biochar and P fertilizer application on growth, yield, and water use efficiency of chickpea. Soil types include Rhodic Ferralsols (clay) in Thohoyandou and Leptic Cambisols (loamy sand) in Nelspruit, South Africa. Treatments consisted of a factorial combination of four biochar levels (0, 5, 10 and 20 t ha^?1) and two phosphorus fertilizer levels (0 and 90 kg ha^?1) arranged in a randomized complete block design and replicated three times. Biochar application at 5 t ha^?1 significantly increased biomass, grain yield and water use efficiency of biomass production (WUE_b) in the clay soil compared to 10 and 20 t ha^?1. However, the increase was attributed to the addition of P fertilizer. Biochar application had no effect on yield components in the loamy sand soil, but P fertilizer addition increased number of seeds/pod in the loamy sand soil and number of pods/plant in the clay soil. Biochar and P fertilizer application on growth and yield of chickpea varied in soil types and seasons, as the effect was more prominent in the clay soil than the loamy sand soil during the summer sowing.