Effect of plant growth promoting rhizobacteria,nematode parasitic fungi and root-nodule bacterium on root-knot nematodes Meloidogyne javanica and growth of chickpea

Effects of plant growth promoting rhizobacteria (Pseudomonas putida MTCC No. 3604 and Pseudomonas alcaligenes MTCC No. 493) and parasitic fungi (Pochonia chlamydosporia KIA and Paecilomyces lilacinus KIA) were studied, alone and together with Rhizobium sp. (charcoal commercial culture) on the growth of chickpea and multiplication of Meloidogyne javanica. Individually, P. putida 3604, P. alcaligenes 493 and Rhizobium caused a significant increase in the growth of chickpea in both nematode inoculated and uninoculated plants. Inoculation of Rhizobium with a parasitic fungus or with plant growth promoting rhizobaterium caused a greater increase in the growth of plants inoculated with nematodes than caused by either of them singly. Individually, P. lilacinus KIA caused a greater increase in the growth of nematode inoculated plants than caused by P. putida 3604 or P. alcaligenes 493. P. lilacinus KIA caused a greater reduction in galling and nematode multiplication followed by P. chlamydosporia KIA, P. putida 3604 and P. alcaligenes 493. Combined use of P. lilacinus KIA with Rhizobium was better in reducing galling and nematode multiplication than any other treatment. P. putida 3604 caused a greater colonization of root than P. alcaligenes 493 while P. lilacinus KIA was isolated from more nematodes than P. chlamydosporia KIA.     

Assessment of the Zn status of chickpea by plant analysis

Chickpea (Cicer arietinum L.) is extensively grown in areas where soils are deficient in zinc (Zn). To determine the response of chickpea to Zn nutrition and to diagnose Zn status in plant tissue, two glasshouse experiments were conducted using Zn-deficient siliceous sandy soil. In Experiment 1, two genotypes of desi chickpea (Dooen and Tyson) were grown at five Zn levels (0, 0.04, 0.2, 1.0 and 5.0 mg kg^-1 of soil). After 4 weeks, no difference in growth and no visible symptoms of Zn deficiency were detected. After 6–8 weeks of growth, chlorosis of younger leaves and stipules occured in the Zn₀ treatment, with shoot dry weight being only 24% of that recorded at the highest Zn level. Root growth increased from 0.52 g/plant when no Zn was applied to 1.04 g/plant in the treatment with 0.2 mg Zn kg^-1 of soil; no response to further increase of Zn fertilization occurred. Zinc concentration in the whole shoot increased significantly with increased in Zn application. The critical Zn concentration in the shoot tissue, associated with 90% of maximum growth, was 20 mg kg^-1 for both genotypes at flowering stage.In the second experiment, two genotypes of desi chickpea (Tyson and T-1587) were grown at three Zn levels (0, 0.5 and 2.5 mg kg^-1 of soil) under two moisture regimes (field capacity 12% w/w, and water stress 4% w/w). Shoot growth was influenced by both Zn supply and water stress. The effect of water stress was severe in the 0.5 and 2.5 mg Zn treatments where shoot dry matter was reduced 52 and 46%, respectively. T-1587 was less sensitive to Zn deficiency and produced higher shoot dry weight than Tyson in the Zn₀ treatment. Zinc concentration in shoots increased from 5 mg kg^-1 when no Zn was applied to 40 mg kg^-1 at the highest Zn level. The critical Zn concentration in shoots was 21 mg kg^-1.The results of the two experiments showed that the critical concentration for Zn did not differ amongst the three cultivars used and was not affected by soil moisture. Similar studies should be undertaken with a wider number of genotypes to discover if a critical concentration of 20–21 mg kg^-1 in the shoot can be used to diagose the Zn status of chickpea genotypes.     

Effects of nitrogen supply and high temperature on the growth and physiology of the chickpea

Chickpeas were grown with or without nitrate nitrogen feeding, or nodulated with Rhizobium leguminosarum. High [40°C day, 25°C night (HT)] and moderate [25°C day, 177°C night (LT)] temperature regimes were employed during growth. Growth rates, photosynthetic capacity and enzymes of carbon and nitrogen metabolism were monitored to assess the acclimatory capacity of the chickpea. Initial growth rates were stimulated by high temperatures, particularly in nitrate-fed and nodulated plants. Older HT plants had fewer laterals, smaller leaves, and fewer flowers were produced than in LT plants. There was some indication of an acclimation of photosynthesis to high temperatures and this was independent of nitrogen supply. Rubisco activity was increased by high growth temperatures. However, HT plants also had higher transpiration rates and lower water use efficiency than LT plants both in respective growth conditions and when compared in a common condition. High temperatures reduced shoot nitrate reductase activity but had little effect on root activity, which was the same if not greater than activity in LT roots. The amino acid, asparagine, was found at high concentrations in all treatments. Concentrations were maintained throughout growth in HT plants but declined with age in LT plants.     

Multiple shoot induction by benzyladenine and complete plant regeneration from seed explants of chickpea (Cicer arietinum L.)

The efficacy of benzyladenine (BA) to induce multiple shoots from seed explants of chickpea (Cicer arietinum L.) was assessed. Shoot differentiation was influenced by the type of seed explant, genotype and concentration of BA. Orientation of the explant also strongly influenced the shoot regeneration response. The optimum BA concentration for shoot/shoot bud regeneration was genotype dependent. Two types of BA-induced response were observed: (1) at less than 7.5 gm BA, direct shoot differentiation (2 to 4-cm-long shoots) was observed within 30 days; (2) at higher BA concentrations (75–100 m), shoot/shoot bud differentiation was achieved in 45–90 days. A high BA concentration inhibited subsequent rooting of shoots. Roots, however, could be easily induced on shoots derived from <12.5 m BA. Following transfer to soil, 80% of the regenerants developed into morphologically normal and fertile plants.Abbreviations BA Benzyladenine     

Studies on the expression of marker genes in chickpea

Conditions were established for the optimum transient expression of beta-glucuronidase and neomycin phosphotransferase II genes introduced into zygotic embryos of chickpea (Cicer arietinum L. 6153 and CM72) by accelerated tungsten particles. Plasmid DNA at a concentration of 12 microgram per milligram of tungsten particles when accelerated with an inflow of helium gas at 60 kilogram per square centimeter through a distance of 24 centimeter in a chamber maintained at a negative pressure of 71.12 centimeter of mercury, resulted in optimal transient expression of the beta-glucuronidase gene in chickpea embryos. However, the expression of the marker genes was 20-40% higher under a cauliflower mosaic virus promoter in comparison to the Win6 and actin promoters. When Agrobacterium tumefaciens was used to transfer marker genes into zygotic embryos and the resultant plants were analysed for activity of the beta-glucuronidase and neomycin phosphotransferase II genes, both of these genes were expressed in tumorous tissues. When a disarmed strain of Agrobacterium was used, normal shoots were regenerated in which the lower parts showed expression of both genes at a frequency of 20%. This revised version was published online in June 2006 with corrections to the Cover Date.     

不同生长调节剂对马蹄金愈伤组织诱导的影响

马蹄金是一种优良的地被兼观赏草坪植物。采用正交设计试验法 ,研究了四种不同生长调节剂对马蹄金子叶、叶片、叶柄和下胚轴愈伤组织诱导的影响。结果表明 :生长调节剂是诱导愈伤组织的关键 ,2 ,4 D对愈伤组织诱导具有显著的影响 ,适宜于马蹄金愈伤组织诱导的培养基及生长调节剂为MS +1 .0mg/L 2 ,4 D +0 .2mg/L 6 BA +0 .2mg/LKT +1 .0mg/Lα NAA。     

Ascochyta blight of chickpea reduced 38% by application of Aureobasidium pullulans (anamorphic Dothioraceae,Dothideales) to post-harvest debris

In 2004–2005, application of non-amended suspensions of Aureobasidium pullulans conidia to post-harvest chickpea debris resulted in 37.9% fewer Ascochyta blight lesions on chickpea test plants relative to controls. Analogous tests in 2006–2007 resulted in 38.4% fewer lesions. Ascospores released from debris were predominantly Davidiella sp. (anamorph, Cladosporium sp.), followed by Didymella rabiei (anamorph, Ascochyta rabiei, agent of Ascochyta blight).     

Effect of plant growth regulators on plant regeneration of Dioscorea remotiflora (Kunth) through nodal explants

Dioscorea remotiflora (Kunth) is an important wild plant that produces tuberous roots used as a source of food in the Western part of Mexico. Lack of planting material and inefficiency of traditional methods of propagation are the main constraints for implementing large-scale cultivation. In contrast, tissue culture techniques allow increasing multiplication and rapid production of plant material. In this regard, leaves or nodal segments were incubated on MS, B5 and WPM culture media with different PGRs in order to obtain an efficient micropropagation protocol. Leaves explants were unable to inducing shoots or callus. However, nodal segments produced axillary shoots and/or callus in all culture media. MS containing 2.33???M KIN was the most suitable to inducing shoots; an average of 6.6 shoots per segment for 100?% explants was obtained, which displayed also the greater number of nodes (5.0) and leaves (7.9) per segment. A decrease on shoot proliferation was observed combining BA or KIN with 2,4-D or NAA. However, small brownish callus were induced on 100?% of segments using 2.33???M KIN with 5.37???M 2,4-D or 9.30???M KIN plus 2.69???M NAA. In contrast, by adding 2.69???M NAA, 66.4?% of the nodal segments formed shoots and produced also yellowish friable callus on the base of the shoots. Shoots were easily rooted with 8.28???M IBA (96.9?%), displaying the greatest root and shoot biomass, but maximum number of tuberous roots, and root or tuberous root biomass was produced increasing IBA (20.7???M).     

Biochemical,physiological and molecular characteristics of chickpea rhizobacteria from Kurdistan province,Iran

Kurdistan province of Iran is one of the main places for producing chickpea, and there is no published research on root-nodulating bacteria of this crop. Plant samples were collected and a total of 73 Rhizobium strains were isolated from root nodules. Nodulation test was done on chickpea plants. Phenotypic characteristics of the 16 representative strains were determined based on the standard bacteriological methods. Total soluble cell protein patterns by electrophoresis approach (SDS-PAGE) showed heterogeneity among the tested rhizobia strains. Based on the phenotypic features, Rhizobium strains of three groups belong to different species of the genus Mesorhizobioum including M. ciceri and M. mediterraneum and Mesorhizobium sp. The PCR technique was employed for amplification of 16S rDNA and atpD genes. For further characterisation, amplified fragment of 16S rDNA gene from a representative strain (AK21) using primers 41F and 1488R was subjected to sequencing. Sequences were aligned by BLAST software at NCBI GenBank and results showed 99% similarity with M. mediterraneum strain BKBCF3q.     

Purification and characterization of an N-acetyl-D-galactosamine-specific lectin from seeds of chickpea (Cicer arietinum L.)

A novel lectin (CAA-II) was isolated and purified from the seeds of Cicer arietinum by ammonium sulphate fractionation and affinity chromatography on an N-acetyl-D-galactosamine-linked agarose column. The lectin is composed of four identical subunits of 30 kDa and the molecular mass of the native lectin was estimated to be 120 kDa by gel filtration chromatography and confirmed by mass spectrometry. The lectin showed agglutination activity against rabbit erythrocytes (trypsin-treated and untreated) as well as against human erythrocytes. Haemagglutination inhibition assays showed that the lectin is a galactose-specific protein having a high affinity for N-acetyl-D-galactosamine. The molecular weight, haemagglutination pattern, carbohydrate specificity and N-terminal amino acid sequence indicated that the lectin is clearly distinct from the previously reported chickpea lectin CAA-I.     

植物生长调节剂对银柴胡细胞生长特性的影响

利用细胞悬浮培养技术,研究不同激素对银柴胡细胞生长特性及过氧化氢酶和硝酸还原酶活性的影响。结果表明,第4种激素配比的培养液M₄(MS+6-BA 1.0 mg·L^-1+2,4-D 1.0 mg·L^-1+NAA 1.0 mg·L^-1)的细胞鲜重、干重,细胞数目均明显高于其他处理;M₄处理可有效增强银柴胡细胞硝酸还原酶的活性,提高氮素利用率;M₂(MS+6-BA 1.0mg·L^-1+NAA0.5 mg·L^-1)处理有利于增强银柴胡细胞过氧化氢酶活性。     

Strains of Mesorhizobium amorphae and Mesorhizobium tianshanense, carrying symbiotic genes of common chickpea endosymbiotic species, constitute a novel biovar (ciceri) capable of nodulating Cicer arietinum

AIMS: To identify several strains of Mesorhizobium amorphae and Mesorhizobium tianshanense nodulating Cicer arietinum in Spain and Portugal, and to study the symbiotic genes carried by these strains. METHODS AND RESULTS: The sequences of 16S-23S intergenic spacer (ITS), 16S rRNA gene and symbiotic genes nodC and nifH were analysed. According to their 16S rRNA gene and ITS sequences, the strains from this study were identified as M. amorphae and M. tianshanense. The type strains of these species were isolated in China from Glycyrrhiza pallidiflora and Amorpha fruticosa nodules, respectively, and are not capable of nodulating chickpea. These strains carry symbiotic genes, phylogenetically divergent from those of the chickpea isolates, whose nodC and nifH genes showed more than 99% similarity with respect to those from Mesorhizobium ciceri and Mesorhizobium mediterraneum, the two common chickpea nodulating species in Spain and Portugal. CONCLUSIONS: The results from this study showed that different symbiotic genes have been acquired by strains from the same species during their coevolution with different legumes in distinct geographical locations. SIGNIFICANCE AND IMPACT OF THE STUDY: A new infrasubspecific division named biovar ciceri is proposed within M. amorphae and M. tianshanense to include the strains able to effectively nodulate Cicer arietinum.     

Active extrusion of protons and exudation of carboxylic acids in response to iron deficiency by roots of chickpea (Cicer arietinum L.)

A chickpea cultivar, K-850, acidified the nutrient solution in response to iron deficiency, with subsequent re-greening of chlorotic leaves. No recovery of chlorosis was observed when the nutrient solution was buffered at a pH 6.3. During the period of acidification induced by iron deficiency, the roots of K-850 exuded more carboxylic acids than when supplied with sufficient iron. However, the rate of extrusion of protons was much higher than the rate of exudation of carboxylic acids during the acidification period. The extrusion of protons was inhibited by the addition of vanadate at the beginning of the decrease in pH. It appeared that acidification of the solution in response to iron deficiency was mediated by a proton-pumping ATPase, located at the plasma membrane. The presence of cations in the solution was essential for the extrusion of protons under iron deficiency, but the species of cation made no significant difference to the rate of extrusion of protons from roots. Therefore, we concluded that non-specific H+/cation antiport was involved in the acidification process.     

Effect of short-chain fatty acids on the ethylene pathway in embryonic axes of Cicer arietinum during germination

The effect of short-chain saturated fatty acids (C₅–C₁₀) on the biosynthesis of ethylene in embryonic axes of chick-pea ( Cicer arietinum L.) seeds was investigated. The emergence of radicle and fresh weight of embryonic axes diminished with increasing number of carbons. The inhibition of germination caused by lower concentrations (1 m M ) of fatty acids (C₅–C₁₀) was partially reversed by exogenous 1-aminocyclopropane-1-carboxylic acid (ACC), whereas exogenous ethylene was able to overcome the inhibitory effect provoked by all concentrations (1–5 m M ) of applied fatty acids (C₅–C₁₀). Ethylene production rates, and enzyme activities of ACC synthase and ACC oxidase decreased concomitantly with the molecular mass and increasing concentration of fatty acids. The inhibitory effect of these acids on ethylene production seems to result not only from a decreased ACC synthesis, but also from an enhancement of 1-malonylamino)cyclopropane-1-carboxylic acid (MACC) synthesis.     

Enhanced plant regeneration in pearl millet (Pennisetum americanum) by ethylene inhibitors and cefotaxime

Cefotaxime, a cephalosporin antibiotic, and different ethylene inhibitors, such as silver nitrate, cobalt chloride, nickel chloride and O-acetyl salicylic acid, significantly delayed the loss of regeneration potential in embryogenic cultures of Pennisetum americanum. In the presence of these chemicals, ethylene content in the atmosphere of the culture vessel was less than that of the control. Cefotaxime, silver nitrate and O-acetyl salicyclic acid did not have any effect on callus growth based on fresh weight, while growth based on dry weight was enhanced by O-acetyl salicyclic acid.Abbreviations ASA O-acetyl salicylic acid - BA benzyladenine - CW coconut water - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - NAA -naphthaleneacetic acid - MS Murashige and Skoog     

Morphological and pathogenic variability of Indian isolates of Fusarium oxysporum f. sp. ciceris causing chickpea wilt

Wilt of chickpea (Cicer arietinum) caused by Fusarium oxysporum f. sp. ciceris is prevalent in almost all chickpea growing areas of the world and its incidence varied from 14.1 to 32.0% in the different states of India surveyed. The isolates were highly variable in their colony growth pattern, size of colony and pigmentations. The size of microconidia varied from 5.1–12.8 × 2.5–5.0 μm, whereas macroconidia ranged from 16.5–37.9 × 4.0–5.9 μm with 1–5 septations. One hundred and twelve isolates were grouped into 12 categories on the basis of their radial growth, size of macroconidia and growth pattern. Majority of the isolates were highly pathogenic causing more than 50% wilt in chickpea cultivar JG 62. Virulence analysis of 56 representative isolates on a set of 18 cultivars of chickpea, including 10 international differentials, grouped them into three categories. Chickpea cultivar KWR 108 differentiated all isolates of Punjab, Haryana and Delhi states and a few isolates of Rajasthan from others by showing resistant reactions and were placed in the first group. The rest of the isolates of Rajasthan state showed susceptible reactions on KWR 108 placed in a second group. Cultivar CPS 1 distinguished the isolates of Jharkhand state and placed them into a third group. An international set of cultivars recommended for race differentiation were not able to distinguish all the isolates into known races of the pathogen, therefore cultivar KWR 108 should be included in the existing differential set of cultivars.     

Effect of plant growth regulators on the natural and ethylene induced pigmentation in Kinnow mandarin peels

Following harvest, Kinnow mandarin (Citrus nobilis × Citrus deliciosa) fruits were variously treated with gibberellins (GA) and cytokinins. Ethylene caused marked chlorophyll (Chl) degradation and its effect was partially reversed by kinetin and benzylaminopurine (BAP) and to a lesser extent by GA₃ and GA₄₊₇. No appreciable accumulation of carotenoids (Car) occurred in these fruits irrespective of treatment. The loss of Chl during natural maturation was significantly reduced by cytokinins. Treatments with gibberellins alone or in combination with cytokinins were much less effective. The reduced loss of Chl in response to exogenously applied cytokinins may be probably related to a decline in its endogenous levels.     

Effects of explants and growth regulators in garlic callus formation and plant regeneration

We intended to evaluate the effects of different explants and growth regulators on callus induction and plant regeneration in garlic (Allium sativum L.). Furthermore, we intended to differentiate among different morphological types of callus by light microscopy and to relate them with their abilities to regenerate plants in the red-garlic cultivar 069. A factorial design with BDS—basal Dunstan and Short (1977)—medium, as a control and supplemented with 0.042, 0.42 and 4.24 μM picloram or with 0.045, 0.45 and 4.5 μM 2,4-D, in both cases with and without 4.43 μM N⁶-benzylaminopurine (BAP), was used. The cultures were grown in darkness at 25 ± 2°C and they were subcultured over a 6-month period. Basal plates and meristems were highly responsive explants, while immature umbels and root-tips were less responsive ones, as indicated by percentage of induced callus, growing callus and regenerating callus. The best response was 41% regenerating callus with 0.045 μM 2,4-D and BAP from basal plates while 57, 56 and 20% regenerating callus were obtained with 0.45 μM 2,4-D from meristems, root-tips and immature umbels, respectively. Also, these treatments showed a higher percentage of nodular and embryogenic callus (type I). Thus, it can be concluded that the use of meristems and 2,4-D will enhance callus production and quality, increase plant regeneration and allows to develop a protocol suitable for further transformation experiments in garlic.     

Enhancement of shoot regeneration from cotyledon explants of Brassica rapa ssp oleifera through pretreatment with auxin and cytokinin and use of ethylene inhibitors

The presence of benzyladenine or naphthaleneacetic acid in seed germination medium markedly enhanced subsequent shoot regeneration from the base of the excised cotyledon explants of Brassica rapa cv. Horizon. Cotyledon explants from younger seedlings (3 or 4-day old) produced more shoots than those from older seedlings. Addition of the ethylene inhibitor aminoethoxyvinylglycine (1.0 M) to the regeneration medium improved shoot regeneration three fold.Abbreviations AVG aminoethoxyvinylglycine - BA benzyladenine - MGBG methylglyoxal-bisguanylhydrazone - MSBN ms (murashige & skoog 1962) medium supplemented with 4.4 m BA & 5.4 m NAA, 2% sucrose - NAA naphthaleneacetic acid