Caecal fermentation characteristics, blood composition and growth of rabbits on substitution of soya-bean meal by unconventional high-glucosinolate mustard ( Brassica juncea) meal as protein supplement

Effect of graded levels of high-glucosinolate mustard (Brassica juncea) meal as substitute of soya-bean meal (SBM) in broiler rabbit diets was studied. Forty weaning rabbits of Soviet Chinchilla and White Giant breed were randomly allocated to one of four experimental diets containing mustard meal (MM) 0, 80, 160 and 245 g/kg. The experiment lasted for 8 weeks. MM had 54.8 mg total glucosinolates (TGLSs) per g dry matter (DM). Diets had TGLS 3.8, 8.4 and 11.98 mg/g DM in 80, 160 and 245 g MM diets, respectively. MM-incorporated diets had higher digestible and linearly (P < 0.01) higher metabolisable energy (ME) content. However, the effect on total tract apparent digestibility of DM, and crude protein was quadratic. Average daily gain (ADG) reduced (P < 0.05) linearly with increasing MM levels in diet, still 80 and 160 g MM diets had similar ADG compared to that of SBM diet. Caecum weight reduced linearly (P < 0.05) with increasing MM levels in diet. The pH of caecal content ranged between 5.85 and 6.19, total N between 1.19 and 1.48 (g per 100 g) and total volatile fatty acids between 4.7 and 5.8 mmol per 100 g, and they were not statistically different. NH3-N ranged between 31.2 and 39.0 mg per 100 ml, and reduced linearly (P < 0.05) while trichloroacetic acid-precipitable nitrogen increased linearly (P < 0.01, ranged between 114 and 247 mg per 100 ml) with increasing MM levels in diet. Blood haemoglobin, packed cell volume and lymphocytes were higher (quadratic effects, P < 0.05) on 245 MM diet, whereas white blood cell count reduced linearly (P < 0.01). Serum aspartate aminotransferase increased linearly (P < 0.01) while alanine aminotransferase and alkaline phosphatase activity, protein, erythrocytes sedimentation rate and red blood cell counts were not affected by MM. Serum Cu, Na and K content increased linearly (P < 0.05) with increasing MM levels. Liver Cu concentration showed quadratic (P < 0.05) increase. Rabbits tolerated 8.4 mg TGLS per g diet (160 g MM per kg) during active growth without any apparent effect on health and growth. It is concluded that MM can replace up to 66% SBM protein in rabbit feeding, whereas complete replacement of SBM with MM reduced feed intake and ADG by 23% and 13%, respectively. Further studies are required to confirm these inclusion levels and glucosinolate tolerance of rabbits.     

Effect of sodium dodecyl sulphate on the high molecular weight protein fraction of mustard (Brassica juncea) and rapeseed (Brassica campestris)

The effect of sodium dodecyl sulphate on mustard and rapeseed 12S protein has been monitored by the techniques of ultracentrifugation, viscosity, difference spectra and fluorescence spectrophotometry. At low concentration of sodium dodecyl sulphate (<3.47 mM) mustard protein undergoes aggregation and at higher concentrations it dissociates to 1.8 S protein, the dissociation being complete at 17.3 mM sodium dodecyl sulphate. The rapeseed protein, on the other hand, undergoes dissociation at all the concentrations of sodium dodecyl sulphate. The reduced viscosity values of mustard protein in the presence of the denaturant are higher than those of rapeseed protein. Similarly in difference spectra change in absorbance values of mustard protein are higher.’ The relative fluorescence intensity of the mustard protein increases with sodium dodecyl sulphate concentration, upto 0.87 mM and this is followed by fluorescence quneching at higher denaturant concentrations. However, with the rapeseed protein fluorescence quenching was observed at all concentrations of sodium dodecyl sulphate.     

Effect of ‘Mixtalol’ on growth,yield and yield components of Indian mustard (Brassica juncea)

Mixtalol (a mixture of long chain aliphatic alcohols varying in chain length from C₂₄ to C₃₂) applied to Brassica juncea plants as foliar spray caused an increase in secondary and tertiary branching with consequent enhancement in seed yield through increased number of inflorescences and siliquae per plant. The percentage of immature siliquae and shattering of siliquae decreased with this treatment. Mixtalol increased total dry matter of plants, partitioning coefficient and harvest index. The contents of starch, protein and oil were also higher in seeds from Mixtalol treated plants.     

Denaturation of the high molecular weight protein fraction of mustard (Brassica juncea) and rapeseed (Brassica campestris) by urea or guanidinium hydrochloride

Urea and guanidinium hydrochloride dissociate the 12S protein of mustard and rapeseed to 1.8 S protein and the extent of dissociation depends on the concentration of the denaturant. Mustard (Brassica juncea) protein is more readily dissociated than the rapeseed (Brassica campestris) protein. The reagents denature the protein as evidenced by increase in viscosity, appearance of difference spectra and quenching of fluorescence. Rapeseed protein is denatured more readily than the mustard protein. Analysis of visctosity, spectral and fluoresence data suggests that the first event in the denaturation reaction is the perturbation of the aromatic amino acid residues followed by their exposure to the solvent medium and unfolding of the protein molecule.     

Fine mapping of loci involved with glucosinolate biosynthesis in oilseed mustard (Brassica juncea) using genomic information from allied species

Fine mapping of six seed glucosinolate QTL (J2Gsl1, J3Gsl2, J9Gsl3, J16Gsl4, J17Gsl5 and J3Gsl6) (Ramchiary et al. in Theor Appl Genet 116:77–85, 2007a) was undertaken by the candidate gene approach. Based on the DNA sequences from Arabidopsis and Brassica oleracea for the different genes involved in the aliphatic glucosinolate biosynthesis, candidate genes were amplified and sequenced from high to low glucosinolate Brassica juncea lines Varuna and Heera, respectively. Of the 20 paralogues identified, 17 paralogues belonging to six gene families were mapped to 12 of the 18 linkage groups of B. juncea genome. Co-mapping of candidate genes with glucosinolate QTL revealed that the candidate gene BjuA.GSL-ELONG.a mapped to the QTL interval of J2Gsl1, BjuA.GSL-ELONG.c, BjuA.GSL-ELONG.d and BjuA.Myb28.a mapped to the QTL interval of J3Gsl2, BjuA.GSL-ALK.a mapped to the QTL interval of J3Gsl6 and BjuB.Myb28.a mapped to the QTL interval of J17Gsl5. The QTL J9Gsl3 and J16Gsl4 did not correspond to any of the mapped candidate genes. The functionality and contribution of different candidate genes/QTL was assessed by allelic variation study using phenotypic data of 785 BC₄DH lines. It was observed that BjuA.Myb28.a and J9Gsl3 contributed significantly to the base level glucosinolate production while J16Gsl4, probably GSL-PRO, BjuA.GSL-ELONG.a and BjuA.GSL-ELONG.c contributed to the C3, C4 and C5 elongation pathways, respectively. Three A genome QTL: J2Gsl1harbouring BjuA.GSL-ELONG.a, J3Gsl2 harbouring both BjuA.GSL-ELONG.c and BjuA.Myb28.a and J9Gsl3, possibly the ‘Bronowski genes’, were identified as most important loci for breeding low glucosinolate B. juncea. We observed two-step genetic control of seed glucosinolate in B. juncea mainly effected by these three A genome QTL. This study, therefore, provides clues to the genetic mechanism of ‘Bronowski genes’ controlling the glucosinolate trait and also provides efficient markers for marker-assisted introgression of low glucosinolate trait in B. juncea. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effect of organic zinc supplementation on growth, nutrient utilization and mineral profile in lambs

To study the efficacy of organic zinc (Zn) supplementation on growth, nutrient utilization and mineral profile as compared to inorganic source [zinc sulphate (ZnSO₄)], 18 Muzaffarnagari male lambs of 11.30 ± 0.45 kg mean body weight (4–5 months of age) were divided into three groups of six animals in each in a randomized block design. Lambs in the control group were fed a standard total mixed ration (TMR) consisted of 60 kg/100 kg of concentrate mixture (CM) and 40 kg/100 kg of wheat straw. CM was consisted of 300 g/kg crushed maize grain, 270 g/kg soybean meal, 400 g/kg wheat bran, 20 g/kg mineral mixture (without Zn) and 10 g/kg common salt. Animals in the experimental groups were additionally supplemented with 20 mg Zn/kg of diet either through inorganic (ZnSO₄) or organic [Zn-methionine AA complex (Zn-meth)] sources. Experimental feeding was done for a period of 150 days including a 6 days metabolism trial. The intake of dry matter (DM), organic matter (OM), crude protein (CP), digestible CP and total digestible nutrients and digestibility of DM, OM, CP, ether extract, neutral detergent fibre and hemicellulose were comparable (P>0.05) among the three groups. However, digestibility of cellulose and acid detergent fibre was significantly (P<0.05) higher in Zn-meth group as compared to control group. Though the balance of calcium was adversely affected (P<0.01) in both the Zn supplemented groups, but it was significantly higher in Zn-meth group compared to ZnSO₄ group. While apparent absorption and retention of nitrogen, phosphorus, copper, iron and manganese were similar (P>0.05) among different groups, retention of Zn (P<0.05) as well as its concentration in the serum (P<0.01) were highest in Zn-meth group, followed by ZnSO₄ group and lowest in the control group, suggesting higher bioavailability of Zn from Zn-meth as compared to ZnSO₄. Average daily gain of the lambs and feed conversion efficiency were also significantly (P<0.05) higher in Zn-meth group as compared to control and ZnSO₄ groups, suggesting a positive role of organic zinc supplementation on the performance of lambs.     

Genetic analysis of agronomic and quality traits in mustard (Brassica juncea)

To develop an efficient mustard (Brassica juncea) breeding programme, a better knowledge of the genetic control and relationships of the main selected characters is needed. Thus, doubled haploid (DH) lines were evaluated over 2 years in the field. Days to flowering, plant height, thousand-seed weight, fatty acid composition, seed oil content, sinigrin, gluconapin and total glucosinolate contents were determined in the DH population. The influence of seed coat colour was estimated. Results showed significant differences between yellow and brown seeds only for oil and fatty acid contents. Molecular analysis revealed that seed coat colour is associated with two Mendelian trait loci: Bjc1 [on linkage group (LG) 3] and Bjc2 (on LG6). The quantitative trait loci associated with characters—detected by composite interval mapping—were not co-localised and revealed a genetic independence. The results obtained in this study show that the most important agronomic and quality traits of brown mustard could be bred independently. Correlation between the studied traits is also discussed.     

Effect of different levels of selenium supplementation on growth rate, nutrient utilization, blood metabolic profile, and immune response in lambs

Eighteen male lambs (8-9 months of age, 25.00 +/- 0.90 kg body weight) were divided into three groups of six animals in each and fed a total mixed ration (TMR) containing concentrate mixture (30% maize grain, 27% soybean meal, 40% wheat bran, 2% mineral mixture, and 1% common salt) and wheat straw in 65:35 ratio and supplemented with selenium (Se) as sodium selenite at 0 (T1, control), 0.15 (T2), and 0.30 ppm (T3) levels. Experimental feeding was done for a period of 90 days including a 6-day metabolism trial. To assess the growth performance, lambs were weighed every 15 days throughout the experimental period. All the lambs were intramuscularly inoculated with a single dose (2 ml) of haemorrhagic septicaemia oil adjuvant vaccine on 0 day to evaluate the humoral immune response. Blood samples were collected on 0 day and thereafter at 30 days interval. Results revealed that supplementation of Se both at 0.15 and 0.30 ppm levels had no significant (P > 0.05) effect on intake and digestibility of dry matter, organic matter, crude protein (CP), ether extract, neutral detergent fiber, acid detergent fiber, and hemicellulose; balances of calcium and phosphorus; and level and intake of digestible CP and total digestible nutrients. Se supplementation also had no significant (P > 0.05) effect on the levels of serum total cholesterol, total protein, albumin, globulin, albumin/globulin ratio, tri-iodothyronine (T(3)), thyroxine (T(4)), and T(4)/T(3) ratio; and serum glutamate oxaloacetate transaminase and serum glutamate pyruvate transaminase enzyme activity in the lambs. However, there was a significant (P < 0.05) increase in the plasma Se levels, red blood cell glutathione peroxidase enzyme activity, and humoral immune response in both the Se-supplemented groups. Feed (TMR) required per kilogram gain was less by 11.1% and 16.5% in groups T2 and T3, respectively, as compared to control (T1) group. Average daily gain was highest (108.5 g) in group T3, followed by group T2 (98.2 g), and lowest (89.06 g) in the control group (T1). These results indicated that supplementation of 0.15 and 0.3 ppm Se in the diet (having 0.19 ppm Se) of lambs significantly improves their immune response and antioxidant status.     

Chlorophyllase and peroxidase activity during degreening of maturing canola (Brassica napus) and mustard (Brassica juncea) seed

Chlorophyllase and peroxidase activities were measured in relation to seed maturation and degreening in canola ( Brassica napus cvs Westar and Alto) and mustard ( Brassica juncea cvs Cutlass and Lethbridge 22A). Samples of seed collected at the same moisture content were pooled, then divided and used for each assay. During maturation the green pigment (chlorophyll and related pigments) content of canola seed decreased linearly and was lower than that measured in mustard at all moisture contents studied, except for the highest and lowest moisture contents. Chlorophyllides and pheophorbides were detected in canola and were essentially absent in mustard. This difference in accumulation of dephytylated pigments infers differences in the pigment degradation pathways in Brassica species. Interspecific differences in the enzymology of degreening were found. Green pigment degradation was associated with increased chlorophyllase activity and low peroxidase activity in canola and low Chlorophyllase and high perosidase activity in mustard. The possible role of ethylene in seed degreening is discussed.     

The application of ethephon (an ethylene releaser) increases growth, photosynthesis and nitrogen accumulation in mustard (Brassica juncea L.) under high nitrogen levels

Ethephon (2-chloroethyl phosphonic acid), an ethylene-releasing compound, influences growth and photosynthesis of mustard (Brassica juncea L. Czern & Coss.). We show the effect of nitrogen availability on ethylene evolution and how this affects growth, photosynthesis and nitrogen accumulation. Ethylene evolution in the control with low N (100 mg N kg(-1) soil) was two-times higher than with high N (200 mg N kg(-1) soil). The application of 100-400 microl x l(-1) ethephon post-flowering, i.e. 60 days after sowing, on plants receiving low or high N further increased ethylene evolution. Leaf area, relative growth rate (RGR), photosynthesis, leaf nitrate reductase (NR) activity and leaf N reached a maximum with application of 200 microl x l(-1) ethephon and high N. The results suggest that the application of ethephon influences growth, photosynthesis and N accumulation, depending on the amount of nitrogen in the soil.     

Phytoextraction of soil cadmium and zinc by microbes-inoculated Indian mustard (Brassica juncea)

Abstract

A pot experiment was carried out to evaluate the effect of Pseudomonas fluorescens and Trichoderma harzianum inoculation on the uptake of zinc (Zn) and cadmium (Cd) by Indian mustard (Brassica juncea) from the soil having three different concentrations of Zn (300, 600, 900 mg/kg) and Cd (5, 10, 15 mg/kg) separately. Microbial inoculation resulted in significantly better plant growth, available metal content and their uptake than control (without microbes). Available Zn was enhanced, ca.1.6- and 1.4-fold and Cd ca. 2.5- and 1.8-fold, by P. fluorescens and T. harzianum, respectively. P. fluorescens resulted in an increase in Zn uptake by 113.9, 51.9 and 58.4% and T. harzianum by 42.6, 32.1 and 33.9% over control from soils having 300, 600 and 900 mg Zn, respectively, while of the corresponding results for Cd were 110.2, 48.9 and 58.1% with P. fluorescens and 42.6, 30.9 and 33.4% with T. harzianum from soil having 5, 10 and 15 mg Cd, respectively, after 90 days of treatment. In general the rate of metal uptake was higher during the initial 30 days and declined later.     

Artificial synthesis of interspecific chimeras between tuber mustard (Brassica juncea) and cabbage (Brassica oleracea) and cytological analysis

Interspecific chimeras between tuber mustard and red cabbage were obtained by in vitro graft-culture method. Before grafting, 6-day-old seedlings of tuber mustard and red cabbage were vertically half-cut and treated with different concentrations of 6-BA and NAA for 1 min, then, they were symmetrically fit together. As a result, sectorial chimeras were initially produced from the united shoot tips. The maximum frequency of chimeral bud formation reached 6.33% when the vertical sections of tuber mustard and cabbage were treated with 2 mg/l 6-BA and 1 mg/l NAA. When sectorial chimeras were propagated on MS medium containing 1 mg/l 6-BA, periclinal and mericlinal chimeras gradually developed. Chimeral shoots were rooted on half-strength MS medium containing 0.1 mg/l NAA. The rooted chimeras were acclimatized and transferred to the field for cytological and morphological analysis. The results showed that stomata density in the chimeras was significantly higher than that of their parents, while chloroplast size, starch grain size and number were intermediate between the two parents. The chimeras were further analyzed by flow cytometry, and the results indicated that they contained both sets of parental chromosomes. Moreover, chimeral plants possessed valuable characters from the two parents.     

De novo shoot morphogenesis and plant growth of mustard (Brassica juncea) in vitro in relation to ethylene

Role of ethylene in de novo shoot morphogenesis from explants and plant growth of mustard ( Brassica juncea cv. India Mustard) in vitro was investigated, by culturing explants or plants in the presence of the ethylene inhibitors aminoethoxyvinylglycine (AVG) and AgNO₃. The presence of 20 μ M AgNO₃ or 5 μ M AVG in culture medium containing 5 μ M naphthaleneacetic acid and 10 μ M benzyladenine were equally effective in promoting shoot regeneration from leaf disc and petiole explants. However, AgNO₃ greatly enhanced ethylene production which reached a maximum after 14 days, whereas ethylene levels in the presence of AVG remained low during 3 weeks of culture. The promotive effect of AVG on shoot regeneration was overcome by exogenous application of 25 μ M 2-chloroethylphosphonic acid (CEPA), but AgNO_3-induced regeneration was less affected by CEPA. For whole plant culture, AVG did not affect plant growth, although it decreased ethylene production by 80% and both endogenous levels of 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC by 70–80%. In contrast, AgNO₃ stimulated all 3 parameters of ethylene synthesis. Both AgNO₃ and CEPA were inhibitory to plant growth, with more severe inhibition occuring in AgNO₃. Leaf discs derived from plants grown with AVG or AgNO₃ were highly regenerative on shoot regeneration medium without ethylene inhibitor, but the presence of AgNO₃ in the medium was inhibitory to regeneration of those derived from plants grown with AgNO₃.     

Plant regeneration from stem cortex expiant and protoplast cultures of Brassica juncea (mustard)

Summary Plant regeneration from stem cortex explants of 13 genotypes of Brassica juncea was assessed. Regeneration was strongly affected by genotype, as up to 50.6 shoots were produced per 100 calli of the most responsive line (Blaze), whereas no shoots were obtained from less responsive lines (Zeml, Vniimk351). Blaze was chosen for B. juncea stem cortex protoplast isolation. After one week of culture, 11–14% of the cells had divided, and about 0.002% produced 1–2 mm colonies within 6 weeks. Up to 7% of these colonies gave rise to shoots upon transfer to plant regeneration medium.Abbreviations 2,4-D 2,4-dichlorophenoxyaceticacid - BAP 6-benzylaminopurine - FDA fluorescein diacetate - IAA indole acetic acid - IBA indole butyric acid - 2ip 2-isopentenyl-adenine - NAA 1-naphthaleneacetic acid - TD thidiazuron     

Biodegradation of glucosinolates in brown mustard seed meal (Brassica juncea) by Aspergillus sp. NR-4201 in liquid and solid-state cultures

Aspergillus sp. NR-4201 was assessed by degrading glucosinolates in brownmustard seed meal (Brassica juncea). A liquid culture of the strain, in a medium derived from the meal, produced total degradation of glucosinolates at 32 h. Under these conditions, the glucosinolate-breakdown product, allylcyanide, was formed inculture filtrates. In a plate culture under sterile conditions, the growth of the strain inheat-treated meal media was shown to be effective at 30 °C with 51% moisture,as determined by the measurement of the colony growth rate. On the laboratory scale,solid-state culture under the same conditions gave rise to total glucosinolate degradationwithin 48 h. In comparison, under non-sterile conditions in either heat-treated or nonheat-treated meal samples, the degradations were complete after 60 and 96 h, respectively.In these cases, growth was associated with some out-growths of contaminating fungi,mainly Rhizopus sp. and Mucor sp. The glucosinolate-breakdown product,allylcyanide, was not detected in the solid-state meal-media culture presumably due toevaporative loss from the fermentation matrix.     

Isolation of phytase-producing bacteria from Himalayan soils and their effect on growth and phosphorus uptake of Indian mustard (Brassica juncea)

Phytase-producing bacteria (PPB) is being investigated as plant growth promoting rhizobacteria (PGPR) to improve the phosphorus (P) nutrition and growth of plants grown in soil with high phytate content. Phytate is dominant organic P forms in many soils and must be hydrolyzed to be available for plants. Indian mustard (Brassica juncea) is a plant with economic importance in agriculture and phytoremediation, therefore biotechnological tools to improve growth and environmental stress tolerance are needed. In this study, we isolated and characterized PPB from Himalayan soils and evaluated their effect on growth and P uptake by B. juncea under greenhouse conditions. Sixty five PPB were isolated and based on phytate hydrolysis, three efficient PPB were chosen and identified as Acromobacter sp. PB-01, Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13. Selected PPB showed ability to grow at wide range of pH, temperature and salt concentrations as well as to harbour diverse PGPR activities, such as: solubilization of insoluble Ca-phosphate (193–642 μg ml^?1), production of phytohormone indole acetic acid (5–39 μg ml^?1) and siderophore. Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13 showed 50 and 70 % inhibition of phytopathogen Rhizoctonia solani, respectively. Greenhouse potting assay also showed that the bacterization of B. juncea seeds with Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13 significantly increased the biomass and P content in 30 days old seedlings. This study reveals the potential of PPB as PGPR to improve the growth of B. juncea.     

Distinct uptake of tellurate from selenate in a selenium accumulator, Indian mustard (Brassica juncea)

Tellurium (Te) is widely used in industry because of its unique chemical and physical properties, and has recently become a part of everyday life as a component of phase-change optical magnetic disks. However, the recovery of Te from the environment has not been discussed yet. In this regard, we evaluated the potential use of Indian mustard (Brassica juncea), a selenium (Se) accumulator, for the phytoremediation of Te. The Indian mustard plant was exposed to selenate and tellurate and the concentrations of Se and Te and the chemical species in the plant were determined. The Indian mustard plant accumulated less Te than Se, and the amount of Te accumulated in the plant was approximately 1/69 of that of Se. Although the incorporation of selenate was reduced by increasing sulfate concentration in the medium, the incorporation of Te was not affected by it, suggesting that this plant was able to discriminate tellurate from selenate in the roots. Three Te species were detected in the plant. The major species was tellurate. The other two species were not identical to available Te standards and thus could not be identified. Consequently, the Indian mustard plant is inappropriate for the phytoremediation of Te because it can strictly distinguish tellurate from selenate.     

Photosynthetic activity, pigment composition and antioxidative response of two mustard (Brassica juncea) cultivars differing in photosynthetic capacity subjected to cadmium stress

Photosynthetic performance, contents of chlorophyll and associated pigments, cellular damage and activities of antioxidative enzymes were investigated in two mustard (Brassica juncea L.) cultivars differing in photosynthetic capacity subjected to cadmium (Cd) stress. Exposure to Cd severely restricted the net photosynthetic rate (P(N)) of RH-30 compared to Varuna. This corresponded to the reductions in the activities of carbonic anhydrase (CA) and ribulose-1,5-bisphosphate carboxylase (Rubisco) in both the cultivars. Decline in chlorophyll (Chl) (a+b) and Chl a content was observed but decrease in Chl b was more conspicuous in Varuna under Cd treatments, which was responsible for higher Chl a:b ratio. Additionally, the relative amount of anthocyanin remained higher in Varuna compared to RH-30 even in the presence of high Cd concentration, while percent pheophytin content increased in RH-30 at low Cd concentration. A higher concentration of Cd (100 mg Cd kg(-1) soil) resulted in elevated hydrogen peroxide (H(2)O(2)) content in both the cultivars. However, Varuna exhibited lower content of H(2)O(2) in comparison to RH-30. This was reflected in the increased cellular damage in RH-30, expressed by greater thiobarbituric acid reactive substances (TBARS) content and electrolyte leakage. The enhanced activities of antioxidative enzymes, ascorbate peroxidase (APX), catalase (CAT) and glutathione reductase (GR) and also lower activity of superoxide dismutase (SOD) in Varuna alleviated Cd stress and protected the photosynthetic activity.     

Effect of inoculation ofAzospirillum lipoferum on nitrogen fixation in rhizosphere soil,their association with root,yield and nitrogen uptake by mustard (Brassica juncea)

Summary A microplot field experiment was conducted in the presence or absence of P and N application to evaluate the influence of the seed inoculation of mustard (cv. Baruna T₅₉) withAzospirillum lipoferum on N₂-fixation in rhizosphere, association of the bacteria with the roots and grain yield and N uptake. Inoculation significantly increased the N content in rhizosphere soil particularly at early stage (40 days) of plant growth, which was accompanied by the increased association of the bacteria (A. lipoferum) in rhizosphere soil, root surface washing and surface-sterilized macerated root. A significant increase in grain yield and N uptake was also observed due to inoculation. Application of P particularly at the 20 kg. ha^–1 level further enhanced the beneficial effect ofAzospirillum lipoferum inoculation, while N addition markedly reduced such an effect.