Characterization of Italian chickpea (Cicer arietinum L.) germplasm by multidisciplinary approach

Characterization of 15 chickpea (Cicer arietinum L.) accessions (6 cultivars, 3 selected lines and 6 landraces) was performed using morphological, agronomic, and technological traits, together with biochemical (seed storage proteins) and molecular markers. Field trials, performed in two different geographical areas of Southern Italy, one in a plain area near Tyrrhenian sea and one in an hilly internal area, revealed a high influence of field location on yield. A wide variation was also registered for technological traits and molecular markers. Microsatellite (SSR) markers grouped chickpea accessions in different clusters, providing useful information on genetic variation and relationships among them, with polymorphic fragments useful to discriminate among all accessions. On the contrary, seed storage protein pattern showed scarce variation resulting very similar among all the accessions considered.     

Endogenous organic acid variations in different chickpea (Cicer arietinum L.) genotypes

Abstract

Changes in cropping systems during the past century have led to selective pressure on weed flora. Species and ecotypes with characteristics enabling them to survive in high-input farmland have increased in numbers, at the cost of plants lacking these characters. Since the 1950s, the perennial weed species Cirsium arvense (L.) Scop. and Sonchus arvensis L. have mainly been controlled by the herbicide group synthetic auxins like MCPA. During recent decades, C. arvense seems to have become less susceptible to MCPA in both Europe and North America but the reasons are unclear.

To study the importance of selective pressure on weed ecotypes, both short- and long-term studies were carried out in Uppsala, Sweden. The first consisted of two growth-characteristic greenhouse experiments. The hypothesis was that ecotypes of C. arvense and S. arvensis from high-input farmland were different and displayed a more competitive growth pattern than did ecotypes from low-input farmland.

The second study was a field experiment with four ecotypes of C. arvense from low-input farmland to study if selective pressure was in force, over a period of six years. The four ecotypes had different growth characteristics and herbicide sensitivity and they were exposed to crop competition and MCPA treatments during the experimental period. The hypothesis was that ecotypes with a more competitive growth pattern and MCPA tolerance would survive to a greater extent than would other ecotypes. For C. arvense, the results from the growth-characteristic experiment showed that the growth pattern of ecotypes from high-input farmland differed, showing a more directly elongated growth pattern with fewer spines on the leaves compared with ecotypes from low-input farmland, which usually were of rosette-type. Results from the field experiment with C. arvense showed that after six years MCPA-sensitive and/or rosette-type ecotypes had almost disappeared while ecotypes with a more directly elongated growth pattern and less sensitive to MCPA survived to a much greater extent. The conclusion was therefore that when exposed to selective pressure like crop competition and herbicide treatments, ecotypes of C. arvense with a more directly elongated growth pattern and less sensitive to herbicide treatment survived to a greater extent compared with ecotypes missing these traits. Ecotypes from high-input farmland had generally fewer leaf spines than did ecotypes from low-input farmland. This may suggest a trade-off between spine formation and rapid competitive growth. In the growth-characteristic experiment with S. arvensis, no differences between ecotypes from high- and low-input farmland regarding growth characteristics or leaf spines could be detected. This might partly be due to a lower exposure of S. arvensis to selective pressure compared with C. arvense, since S. arvensis generally is less sensitive to MCPA.     

Interaction of chickpea (Cicer arietinum L.) legumin with oxidized linoleic acid.

Chickpea legumin has been purified and incubated under oxidizing conditions with linoleic acid to investigate the influence of this acid on the structure and nutritional quality of the protein. At the end of the incubation time, >30% of the linoleic acid was oxidized. The oxidized linoleic acid was highly detrimental to legumin, and the electrophoretic pattern of the protein was completely changed after the incubation period. Nevertheless, neither polymerization nor cleavage of the protein was observed as deduced from gel filtration chromatography, suggesting that the changes observed in native electrophoresis were probably due to oxidation of legumin. The incubation of legumin with linoleic acid also produced a diminution of the contents of methionine and histidine, by 81.3 and 24.3%, respectively. Finally, in vitro protein digestibility of chickpea legumin was also seriously affected by the incubation with linoleic acid, decreasing from 84.1 to 69.2%.     

Synthesis and quantitative structure-activity relationships of oxanilates as chemical hybridizing agents for wheat (Triticum aestivum L.)

Chemical hybridizing agents (CHAs) can facilitate two-line breeding in heterosis programs of crops. Twenty-seven oxanilates having different aromatic substitutions were synthesized and screened as CHAs on two genotypes of wheat, PBW 343 and HD 2733, during two Rabi (winter) seasons, 2000-01 and 2001-02. The oxanilates prepared by thermal condensation of anilines with diethyl oxalate or by acylation with ethoxycarbonyl methanoyl chloride were sprayed at 1000 and 1500 ppm at the premeiotic stage of wheat, when the length of the emerging spike of the first node was 7-8 mm. Pollen sterility and spikelet sterility were measured in each treatment. Ethyl oxanilates 5, 6, and 25,containing 4-F, 4-Br, and 4-CF(3) aromatic substituents, respectively, induced greater than 98% spikelet sterility, the desired level, at 1500 ppm. Quantitative structure-activity relationship analysis revealed a direct relationship between F(p) and molecular mass but an inverse relationship between MR, E(S), and R in influencing the bioactivity. Several F(1) hybrids were developed using 5, and at least one showed heterosis.     

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     

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.     

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     

Production of extensive chickpea (Cicer arietinum L.) protein hydrolysates with reduced antigenic activity.

Chickpea protein isolate was used as starting material for the production of hypoallergenic protein hydrolysates. Western blotting of the protein isolate showed that IgE in sensitized patient sera strongly bound to the basic polypeptidic chains and recognized the acidic ones of 11S globulin. During the hydrolysis process by the individual and/or sequential action of endo- and exoproteases, a high reduction of antigenic activity was observed. Results suggest that the presence of intact or partially hydrolyzed basic polypeptide chains of 11S globulin are responsible for the formation of IgE complexes in protein hydrolysates obtained by exoprotease treatment; however, the digestion of these polypeptide chains by individual action of endoprotease caused a high loss of antigenic activity. The most effective reduction of antigenicity, >90%, was observed in extensive hydrolyzed chickpea proteins obtained by sequential treatment with endo- and exopeptidases. This chickpea protein hydrolysate could be useful for the elaboration of specialized hypoallergenic food products.     

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.     

Tolerance of chickpea (Cicer arietinum L.) lines to root-knot nematode, Meloidogyne javanica (Treub) Chitwood

The root-knot nematode, Meloidogyne javanica (Treub) Chitwood is an important parasite of chickpea (Cicer arietinum L.). Four chickpea genotypes were evaluated for tolerance to M. javanica in naturally infested fields at three locations. Each genotype was evaluated for number of galls, gall size, root area covered with galls and number of egg masses produced. All the cultivars were susceptible or highly susceptible. Seed yield, weight of 100 undamaged seeds, total dry matter and plant height were compared with checks. Chickpea cultivar Annigeri and a local check were used as nematode susceptible checks in all locations. The four promising nematode tolerant genotypes produced significantly greater yield and total dry matter than the checks in fields naturally infested with M. javanica at three locations. These M. javanica tolerant lines represent new germplasm and they are available in the chickpea genebank at the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) bearing the identification numbers ICC 8932, ICC 11152, ICCV 90043 and ICCC 42.     

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     

Target region amplification polymorphism (TRAP) for assessing genetic diversity and marker-trait associations in chickpea (Cicer arietinum L.) germplasm

The effective utilization of crop diversity held in genebanks depends on our knowledge of useful traits and available markers associated with the target traits. Target region amplification polymorphism (TRAP) was used to evaluate the genetic diversity and underlying relationships among 263 accessions of chickpea landraces maintained by the USDA-ARS Western Regional Plant Introduction Station in Pullman, WA, USA. Two-hundred sixty-two TRAP markers were amplified by eight primer combinations. Altogether, 110 (42 %) markers were polymorphic, the other 152 (58 %) displayed no variation. These polymorphic markers revealed important differences among the accessions, with an estimated, mean pair-wise genetic distance of 25.82 %, ranging from 2.8 to 50.0 %. Genetic distance analysis divided the accessions into two major groups, with 113 and 150 accessions each, and substantial association between molecular diversity and geographic origin was evident. Bayesian analysis of population structure revealed two groups (K = 2) with evidence for six sub-groups. Additionally, the population structure of a subset of 110 lines was determined (K = 3) for testing marker-trait associations (MTAs). Phenotypic traits included the concentrations of protein and nine mineral elements in the seeds. Two MTAs were significant (p < 0.01) for concentrations of Ca and K, and three MTAs were significant for Cu and Ni concentrations. The results indicate that this population is useful for genome-wide association studies on other economic traits given the level of genetic diversity uncovered and the marker-trait associations in seed minerals discovered.     

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.     

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.     

Nutritional responses of rats to diets based on chickpea ( Cicer arietinum L.) seed meal or its protein fractions

The aim of this study was to isolate the protein fractions from chickpea, var. IAC-Marrocos, as well as to evaluate its in vivo nutritional protein quality. Among the proteins, albumins showed better nutritional value in the in vivo assays and amino acid contents, despite their higher trypsin inhibitor contents. Trypsin inhibitors were found to be heat labile in all samples, but the digestibility results for unheated and heated flour and albumins suggest that their contents are not very decisive. The PER values for casein (not supplemented) were very similar to those of heated flour and unheated or heated albumin and total globulins. The albumin and glutelin fractions showed the best results for PDCAAS, however, lower than those of casein. Despite the high digestibility of the globulin the very low essential amino acid content lowered its PDCAAS, and it had the lowest values.     

Stimulation of nodulation and plant growth of chickpea (Cicer arietinum L.) by Pseudomonas spp. antagonistic to fungal pathogens

Two strains of Pseudomonas MRS23 and CRP55b showed antagonistic activity towards the pathogenic fungi Aspergillus sp., Fusarium oxysporum f. sp. ciceri, Pythium aphanidermatum and Rhizoctonia solani under culture conditions. Larger growth inhibition zones were obtained on nutrient agar (NA) and King's B media in comparison to potato dextrose agar and pigment production media. Both the strains produced siderophore in agar plates as well as in liquid cultures. Fungal inhibition zones were reduced in size and abolished in iron-supplemented NA medium by Pseudomonas strains MRS23 and CRP55b, respectively, indicating that some other metabolites along with siderophores are involved in growth inhibition of fungi by strain MRS23, whereas CRP55b produced only siderophores. Only Pseudomonas strain MRS23 was found to produce hydrocyanic acid (HCN). Seed bacterization with Pseudomonas strains of two chickpea (Cicer arietinum L.) cultivars, H8618 and C235, showed root-stunting effects at 5 days, whereas this inhibitory effect was overcome at 10 days of seedling growth in cv. H8618. Coinoculation of chickpea with Pseudomonas strains MRS23 and CRP55b, and Mesorhizobium sp. Cicer strain Ca181 resulted in the formation of 68.2-115.4% more nodules at 80 and 100 days after planting as compared to single inoculation with the Mesorhizobium strain under sterile conditions. The shoot dry weight ratios of coinoculated treatments at different stages of plant growth varied from 1.18 to 1.35 times that of Mesorhizobium-inoculated and 3.25 to 4.06 times those in uninoculated controls. The plant N contents were also increased significantly on coinoculation. Coinoculation effects of HCN-producing strain MRS23 were significantly lower than those of non-HCN-producing strain CRP55b in terms of shoot dry weight and shoot N. The results demonstrated the potential benefits of using rhizosphere bacteria as coinocula in nodule promotion and plant growth in chickpea.     

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.     

Yield enhancement and biofortification of chickpea (Cicer arietinum L.) grain with iron and zinc through foliar application of ferrous sulfate and urea

Abstract

This study investigated effects of iron (Fe) and nitrogen (N) foliar application on Fe and zinc (Zn) content in chickpea grain, grain yield, and protein content. Application of FeSO₄ at 0.5% at flowering?+?pod formation stages resulted in the highest Fe (73.50 and 75.34?mg Fe kg^?1 grain in first and second year) and Zn (35.08 and 34.21?mg Zn kg^?1 grain) content in grain followed by the application of FeSO₄ at 0.5% at flowering stage alone (68.27 and 69.97?mg Fe kg^?1 grain and 32.44 and 32.27?mg Zn kg^?1 grain) and control (54.63 and 55.69?mg Fe kg^?1 grain and 29.48 and 29.07?mg Zn kg^?1 grain). Urea spray at 2% at flowering as well as at flowering?+?pod formation stages also improved the Zn and Fe content in the grain. Combined use of Fe and urea improved the grain Fe and Zn content over sole application of Fe.     

Evaluation of plant growth promoting activities of microbial strains and their effect on growth and yield of chickpea (Cicer arietinum L.) in India

The aims of our study were to enhance growth, yield and disease control of chickpea by various combinations of microbial strains (Mesorhizobium, Azotobacter chroococcum, Pseudomonas aeruginosa and Trichoderma harzianum). Pseudomonas and Trichoderma showed positive IAA (indole-3-acetic acid) production, phosphate solubilisation and antagonistic activities against Fusarium oxysporum and Rhizoctonia solani as compared to other strains. In two year investigations, tetra-inoculants have shown significant growth attributes, yield and phytopathogen growth inhibition followed by tri-inoculants than control. Therefore, tetra-inoculants (Mesorhizobium-Azotobacter-Pseudomonas-Trichoderma) may be used as efficient biofertilizer and bio-control agent for chickpea production (Cicer arietinum L.) in eastern Uttar Pradesh.