Cell membrane stability and biochemical response of cultured cells of groundnut under polyethylene glycol-induced water stress

Cell membrane stability (CMS) in suspension cultures of two groundnut cultivars was studied under polyethylene glycol(PEG)-induced water stress. There was a negative relationship between PEG concentration in the medium and membrane stability measured as electrolyte leakage. The CMS values in the cell cultures correlated well with the whole plant tissue and permitted the differentiation of cultivars based on their known response to drought stress. The cell membrane stability was lower (more electrolyte leakage) when cells were grown in culture as compared to the intact plant tissue. Kadiri-3, the drought tolerant cultivar maintained higher CMS than JL-24, the drought susceptible one. With increasing PEG levels the concentration of Potassium in cultured cells declined in both cultivars. However, Kadiri-3 maintained higher K values than JL-24 accompanied with greater cell membrane stability. Total soluble sugars also increased with increasing stress in both cultivars; the increase being higher in Kadiri-3. There was no significant change in the total free amino acids but proline accumulated markedly in both varieties. However, no relationship was found between proline levels and CMS. The results demonstrated that CMS test can also be used under in vitro conditions to differentiate the drought tolerant and susceptible cultivars and the cellular K level has a positive relationship with membrane stability.     

Evaluation of some fungal diseases and yield of groundnut in groundnut-based cropping systems

A study on the evaluation of some fungal diseases and yield of groundnut in groundnut-based cropping systems was conducted in 2002 and 2003 planting seasons. Analysis of variance indicated that intercropping was highly significant on leaf spot disease severity 0.76; 0.75, rust 2.75; 2.69, as well as percentage defoliation 78.42%; 78.10% in 2002 and 2003, respectively. Plant population was significant on leaf spot severity 4.52, 4.60 rust 2.76; 366 and defoliation 226.5; 441.1 fungal as well as yield, while interactions were not significant on the fungal diseases and yield. Sole groundnut recorded significantly high severity of the fungal diseases investigated but low yield, when groundnut was intercropped with maize and melon and recorded the lowest yield in 2002 and 2003 respectively. 250,000 plants/ha recorded the lowest severity of the diseases investigated, while 444,444 plants/ha recorded the highest. Sole groundnut also recorded the highest percentage defoliation 79.37%, 79.25% when groundnut was intercropped with maize recorded the lowest 77.06%; 77.60%. 250,000 plants/ha had the lowest defoliation when 444,444 plants/ha had the highest 80.75% 82.13% in 2002 and 2003, respectively. Sole groundnut recorded the lowest in the majority of yields when intercropped with maize and with melon recorded a high yield and yield components in 2002 and 2003, respectively. The microorganisms identified were Cercospora spp., Aspergillus linked and Blastomyces.     

Transmission of peanut yellow spot virus (PYSV) by Thrips,Scirtothrips dorsalis Hood in groundnut †

Peanut yellow spot virus (PYSV) was efficiently transmitted by Scirtothrips dorsalis Hood in groundnut. Larvae could acquire the virus in 30 min and the maximum percentage transmission of 43.8% by individual insects resulted following two days AAP. Single adult Thrip transmitted the virus after minimum IAP of 30 minutes. The percentage transmission (33.3%) increased linearly with an increase in IAP up to 1.5 days and maximum up to 55 h of IAP (36.1%). PYSV persistently transmitted more than 75% of their life span.     

Growth and yield of groundnut (Arachis hypogaea L.) plants in soil inoculated with Aspergillus niger Van Tieghem

Aspergillus niger, a soil-borne fungus is a causative agent of hypocotyl malformations in infected groundnut (Arachis hypogaea L.) plants, but its effect on yield is unknown. This study sought to determine its effect on growth and yield. Seeds of Chinese and JL45 varieties were sown in soil inoculated with A. niger. Fresh and dry weights of the shoots and roots were taken at 10-day intervals. Nodule count was done at 30 days after emergence and subsequently at 10-day intervals. Pods of 20 plants each from inoculated and uninoculated soils were harvested. Growth was suppressed in plants grown on A. niger inoculated soil. Eight-day old plants grown in inoculated soil developed curvatures on their hypocotyls. Nodulation was suppressed (p < 0.05) in plants grown in inoculated soil. Although growth was suppressed in plants grown on inoculated soil, yield of both varieties of groundnut was not affected.     

Efficacy of some plant extracts in the management of cercospora leaf spot of groundnut in the Sudan savanna of Nigeria

Field trials were conducted to evaluate the efficacy of neem seed, garlic clove, onion bulb, ginger rhizome and pawpaw leaf extracts applied as foliar spray in controlling cercospora leaf spot of groundnut in the Sudan savanna of Nigeria during the 2002 and 2003 cropping seasons. Ridomil was used as a positive check while the control was left untreated. Three spray regimes (once, twice and thrice) were evaluated. A strip plot design with three replications was used in setting up the experiments. Ex-Dakar, a cercospora leaf spot susceptible Spanish Valencia groundnut variety, was used as planting material. All the plant extracts reduced the incidence and severity of cercospora leaf spot in both seasons compared to the untreated crops. However, neem seed and garlic clove extracts significantly reduced the incidence and severity of the disease compared to the other plant extracts. Ridomil was more effective in controlling the disease than the plant extracts. Three sprays with the plant extracts gave better control of the disease than one or two sprays in the season. The highest seed yield of 3115 kg/ha and 3311 kg/ha in 2002 and 2003, respectively, were obtained with neem seed extract treatment while the lowest seed yield of 2134 kg/ha and 2334 kg/ha in 2002 and 2003, respectively, were recorded from the untreated crops. Also the highest seed yield of 3124 kg/ha and 3418 kg/ha in 2002 and 2003, respectively were obtained following three sprays compared to 2523 kg/ha and 2641 kg/ha for one spray in 2002 and 2003, respectively and 2840 kg/ha and 3085 kg/ha for two sprays in 2002 and 2003, respectively. The highest haulm yield of 4121 kg/ha and 4089 kg/ha in 2002 and 2003, respectively were also obtained following treatment with neem seed extract. The lowest haulm yield of 2951 kg/ha and 3045 kg/ha in 2002 and 2003 respectively, were recorded from the control. Three sprays gave higher haulm yield (4265 kg/ha and 4279 kg/ha in 2002 and 2003, respectively) than one and two sprays in the season. Neem seed extract could therefore be used to control cercospora leaf spot and to improve groundnut production in the Sudan savanna of Nigeria.     

Contamination of Groundnut in South‐Western Nigeria by Aflatoxigenic Fungi and Aflatoxins in Relation to Processing

Groundnut is commonly consumed in its roasted form by many Nigerians. This study was therefore conducted to determine the levels of aflatoxin in roasted groundnut retailed in south‐western Nigeria with a view to assessing the fitness of the processed nut for human consumption. The effects of roasting and de‐coating as alternative methods for reducing the ‘aflatoxin scare’ in the nut were further assessed on aflatoxigenic fungal load and aflatoxin content of the nuts. Forty‐eight samples of retailed raw and roasted groundnut were collected and assessed by mycological and thin‐layer chromatographic analysis for changes in aflatoxigenic fungal population and aflatoxin concentration, respectively. Consequently, 480 isolates of the Aspergillus section Flavi group, A. flavus L strain (n = 410), A. tamarii (n = 56), A. parasiticus (n = 7) and A. parvisclerotigenus (n = 7), were recovered from all samples. Aflatoxigenic isolates of A. flavus L strain (58.8%) had a significantly (P < 0.05) higher incidence than the non‐aflatoxigenic isolates (41.2%). Aflatoxins were detected in 43 (89.6%) of the samples. Approximately 25% of all samples exceeded the 20 ng/g limit for aflatoxin B₁ (AFB₁) adopted by the National Agency for Food and Drug Administration and Control while 83 and 79% of all samples contained AFB₁ and total aflatoxins above the European Union limits of 2 and 4 ng/g, respectively. Aflatoxin concentrations in the raw and coated samples were as much as five times higher than those in the roasted and de‐coated nuts, respectively. However, no significant difference was recorded between aflatoxin levels in the coated and de‐coated samples. This study has shown that roasting of groundnut and testa removal (de‐coating) are effective processing interventions that can significantly lower aflatoxin quantities in the kernels, thus making it fit for human consumption.     

不同类型拌种剂对花生及其根际微生物的影响

拌种处理对于花生一播全苗和稳定高产非常重要.选用不同类型的4种拌种剂即哈茨木霉菌剂(真菌拮抗剂)、甲基托布津(杀真菌剂)、适乐时(杀真菌剂)、好安威(化学杀虫剂)处理花生种子,通过测定花生农艺性状、品质指标、根际土壤微生物动态等来评价其对花生和环境的综合效应.结果表明:1)各种杀菌剂拌种均能提高花生成苗率,而好安威略差;各拌种剂促进花生株高、叶绿素含量,而单株叶片数适乐时增多,好安威减少,单株分枝数盛花期有所增多(好安威除外),结果期均略减少;单株烂、虫、芽果数均减少,单株秕果数增加,单株饱果数降低(好安威除外),百果重和百仁重提高(哈茨木霉除外),而饱满度降低;最终荚果产量除哈茨木霉略低,其余拌种剂增产效果极显著,甲基托布津、适乐时、好安威比对照分别增产34.58％、25.90％、22.82％.2)哈茨木霉拌种使蛋白质含量、亚油酸含量增幅最大,油份含量增幅较大,油亚比值降幅最大;甲基托布津处理的蛋白质和油份含量降幅最大;适乐时对提高油份含量、油亚比值效果最佳;好安威对品质指标的影响有限.3)从细菌及放线菌与真菌的比值来看,甲基托布津促细菌、抑真菌的效果好且长,对放线菌/真菌比值影响较小;哈茨木霉在前中期促细菌与放线菌、抑真菌的效果明显,但后期效果趋反;适乐时促细菌、抑真菌的效果短促,且一直强烈抑制放线菌;而好安威相比影响较小.4)好安威对根瘤菌具有显著的抑制作用,而哈茨木霉、适乐时、甲基托布津均极显著增加根瘤数量.结论:各拌种剂对花生产量、品质、根瘤、根际微生物产生了较大差异的综合效应,须因地、因时选用;化学杀菌剂甲基托布津的农艺效应、环境微生物效应均最好,唯蛋白质含量、油份含量略有降低,是一种较理想的综合优良拌种剂.     

Effect of nontoxigenic Aspergillus flavus and A. parasiticus on aflatoxin contamination of wounded peanut seeds inoculated with agricultural soil containing natural fungal populations

Peanuts and other seed and grain crops are commonly contaminated with carcinogenic aflatoxins, secondary metabolites produced by Aspergillus flavus and A. parasiticus. Aflatoxin contamination of peanuts in the field can be reduced by 77–98% with biological control through the application of nontoxigenic strains of these species, which competitively exclude native aflatoxin-producing strains from developing peanuts. In this study, viable peanut seeds were artificially wounded and inoculated with field soil containing natural fungal populations that were supplemented with conidia of nontoxigenic A. flavus NRRL 21882 (niaD nitrate-nonutilizing mutant) and A. parasiticus NRRL 21369 (conidial color mutant). Increasing soil densities of applied nontoxigenic strains generally resulted in an increase in the incidence of seed colonization by applied nontoxigenic strains, a decrease in seed colonization by native A. flavus and A. parasiticus, and a decrease in aflatoxin concentration in seeds. Reduction of aflatoxins in peanut seeds depended on both the density and the aflatoxin-producing potential of native populations and on the fungal strain used for biological control. Wild-type strain A. flavus NRRL 21882 and its niaD mutant were equally effective in reducing aflatoxins in peanuts, indicating that nitrate-nonutilizing mutants, which are easily monitored in the field, can be used for evaluating the efficacy of biocontrol strains.     

Germplasm Preservation of Wild Arachis Species through Culture of Shoot Apices and Axillary Buds from In Vitro Plants

A study was conducted to evaluate in vitro techniques for germplasm preservation of wild species of Arachis. Nodal segments excised from in vitro-grown plants of A. retusa, A. macedoi and A. burchellii were used to examine the effects of explant position and age of the donor plant. Explants were excised from plants maintained in culture for 30, 60, 90 or 180 d, numbered I – V from top to bottom and cultured on MS medium supplemented with 2.7 µM NAA or different BAP concentrations (0, 4.4, 13.2 and 22 µM). The age of the donor plant has not influenced the responses of the four genotypes studied. In contrast, shoot regeneration ability was significantly affected by the original explant position, decreasing from top to bottom. In media supplemented with different BAP concentrations, multishoot formation was induced from apical segments at low frequencies (10 – 20%) and segments of all positions originated calluses at the explant basis after 30 d of culture. The culture of nodal segments in the presence of 2.7 µM NAA as the sole growth regulator is recommended for the multiplication of in vitro collections of wild groundnut species in order to avoid callusing and adventitious shoot formation.     

Effect of Inoculum Rate of Biological Control Agents on Preharvest Aflatoxin Contamination of Peanuts

Studies were conducted during 1994 and 1995 in the environmental control plot facility at the National Peanut Research Laboratory to determine the effect of different inoculum rates of biological control agents on preharvest aflatoxin contamination of Florunner peanuts. Biocontrol agents were nontoxigenic color mutants ofAspergillus flavusandAspergillus parasiticusthat were grown on rice for use as soil inoculum. Three replicate plots (4.0 × 5.5 m) were treated with 0, 2, 10, and 50 g/m of row (0, 20, 100, and 500 lb/acre, respectively) of an equal mixture of the color mutant-infested rice in 1994, and the same plots were retreated in 1995. Aflatoxin concentrations were determined by high performance liquid chromatographic analysis of all peanuts. Treatment means for total kernels in 1994 were 337.6, 73.7, 34.8, and 33.3 ppb for the 0, 2, 10, and 50 g/m treatments, respectively. Regression analysis indicated a trend toward lower aflatoxin concentrations with increasing rates of inoculum (R²= 0.40;P< 0.05). For the same repeated treatments in 1995 aflatoxin concentrations in total kernels averaged 718.3, 184.4, 35.9, and 0.4 ppb. Regression analysis revealed a stronger relationship between inoculum rate and aflatoxin concentrations (R²= 0.66;P< 0.05) in the second year of treatment. Compared with untreated controls, the 2, 10, and 50 g/m treatments produced respective reductions in aflatoxin of 74.3, 95.0, and 99.9% in the second year. The data indicated not only a treatment-related effect, but also that a higher degree of control might be achieved when plots or fields are retreated with biocontrol agents in subsequent years.