Production of genetically uniform plants from shoot tips of Aconitum heterophyllum Wall. – a critically endangered medicinal herb

We report the successful micropropagation of a critically endangered medicinal plant Aconitum heterophyllum Wall., using low concentrations of plant growth regulators (PGRs) and molecular validation of the clonal stocks. The maximum rate of in vitro shoot multiplication was obtained on 1.0 × Murashige and Skoog (MS) medium containing 0.25 mg L^?1 Kinetin (Kn) plus 0.25 mg L^?1 Indole acetic acid (IAA). Up to 100% rooting was obtained 15 for shoots cultured on 1.0 × MS medium supplemented with 1.0 mg L^?1 IAA. Adding 0.25 mg L^?1 2,4-dichlorophenoxyacetic acid (2, 4-D) to 1.0 × MS medium resulted in 100% callus formation, while adding 0.25 mg L^?1 IAA plus 0.25 mg L^?1 Kn to 1.0 × MS medium containing 0.25 mg L^?1 2,4-D resulted in 100% generation of embryogenic callus. Inter-simple sequence repeat (ISSR) marker analysis was carried out to check for possible somaclonal variation in the plantlets obtained after three consecutive sub-cultures. Of the 15 ISSR primers used, 10 were found to be monomorphic, with 95–98% similarity, and were used for cluster analysis by the unweighted pair group using arithmetic averages (UPGMA) method. The results revealed that in vitro-regenerated plantlets did not exhibit any genetic polymorphism.     

Allelopathic effects of Leucaena leucocephala leaf litter on some forest and agricultural crops grown in nursery

An experiment was conducted to assess the effect of leaf litter of Leucaena leucocephala on two forest crops Sada koroi (Albizia procera), Ipil ipil (L. leucocephala) and three agricultural crops Falen (Vigna unguiculata), Chickpea (Cicer arietinum) and Arhor (Cajanus cajan) in the nursery of the Institute of Forestry and Environmental Sciences, Chittagong University, Bangladesh, in a Randomized Block Design. Results suggested that leaf litters of L. leucocephala induced inhibitory effects on germination and growth of bioassay. It was also found that the effect depended on concentration of extract and litterfall, type of receptor species. Higher concentration of the materials had the higher effect and vice versa. Growth response of receptor crops varied with the variation of leaf litter application. The study revealed that application of low-dose leaf litter specially litter of 10 g·m⁻² had stimulating effect on shoot growth of C. arietinum, V. unguiculata and A. procera. While in all other cases significant inhibitory effect was observed and it was significantly increased with the increase of leaf litter application. However, the trend of inhibition was uneven with treatments. Root growth was found to be more affected than shoot growth.     

Tuberculosis seroprevalence and comparison of hematology and biochemistry parameters between seropositive and seronegative captive Asian elephants of Nepal

We conducted a tuberculosis (TB) serosurveillance program of captive elephants in Nepal and compared hematology and biochemistry parameters between seropositive and seronegative elephants. A total of 153 elephants (male=20, female=133) from four national parks were tested for TB using the ElephantTB STAT-PAK^® Assay (ChemBio Diagnostic Systems, Inc., Medford, NY, USA). The mean reported age for 138 elephants was 38.5 years (range 2–71 years). Seroprevalence for TB was 21.56% (33/153). The majority of seropositive elephants were female (n=30) and from Chitwan National Park (n=29). The occurrence of TB seropositive cases in other more remote national parks suggests TB may be widespread among the captive elephant population of Nepal. Hematology and biochemistry analyses were performed on 13 and 22 seropositive elephants, respectively and, nine elephants from a seronegative TB herd for comparison. Hematology parameters (hemoglobin, packed cell volume, platelet, white blood cells, and erythrocyte sedimentation rate) were comparable between the two groups. Total protein, globulin, and lactate dehydrogenase were significantly higher in seronegative elephants, and bilirubin was significantly higher in seropositive elephants whereas blood urea nitrogen, creatinine, glutamic oxaloacetic transaminase/aspartate aminotransferase (GOT/AST), glutamic pyruvic transaminase/alanine aminotransferase (GPT/ALT), gamma glutamyl transferase (GT), and albumin were not significantly different. The range of biochemical parameters that were significantly different between seropositive and seronegative elephants had narrow ranges. Thus, the potential of these parameters as a direct biomarker for TB diagnosis is limited based on the findings in this study. We recommend including blood parameters in future TB surveillance studies.     

Biological control of sclerotinia stem rot of canola using antagonistic bacteria

Stem rot caused by Sclerotinia sclerotiorum is a major fungal disease of canola worldwide. In Australia the management of stem rot relies primarily on strategic application of synthetic fungicides. In an attempt to find alternative strategies for the management of the disease, 514 naturally occurring bacterial isolates were screened for antagonism to S. sclerotiorum. Antifungal activity against mycelial growth of the fungus was exhibited by three isolates of bacteria. The bacteria were identified as Bacillus cereus (SC‐1 and P‐1) and Bacillus subtilis (W‐67) via 16S rRNA sequencing. In vitro antagonism assays using these isolates resulted in significant inhibition of mycelial elongation and complete inhibition of sclerotial germination by both non‐volatile and volatile metabolites. The antagonistic strains caused a significant reduction in the viability of sclerotia when tested in a greenhouse pot trial with soil collected from the field. Spray treatments of bacterial strains reduced disease incidence and yielded higher control efficacy both on inoculated cotyledons and stems. Application of SC‐1 and W‐67 in the field at 10% flowering stage (growth stage 4·00) of canola demonstrated that control efficacy of SC‐1 was significantly higher in all three trials (over 2 years) when sprayed twice at 7‐day intervals. The greatest control of disease was observed with the fungicide Prosaro^® 420SC or with two applications of SC‐1. The results demonstrated that, in the light of environmental concerns and increasing cost of fungicides, B. cereus SC‐1 may have potential as a biological control agent of sclerotinia stem rot of canola in Australia.     

含羞草叶子提取物对常见农作物种子萌发生长的抑制作用

观察了含羞草叶子提取物对孟加拉常见农作物种子萌发生长的抑制作用.实验在27-30℃的室温和24小时的光照条件下进行.结果表明：不同浓度的含羞草叶子提取物明显影响所选的植物种子根,茎和侧根的延长和生长.生物学测定表明：高浓度（50%-100%）的提取物有较强的抑制作用.而低浓度 （10%-25%） 的提取物的抑制作用较弱.研究还显示,提取物对植物根和侧根的抑制作用比对种子萌发和茎的生长抑制作用更显著.     

Impact of wheat-Leymus racemosus added chromosomes on wheat adaptation and tolerance to heat stress

Adaptation of wheat (Triticum aestivum L.) to high temperatures could be improved by introducing alien genes from wild relatives. We evaluated the responses of wheat-Leymus racemosus chromosome introgression lines to high temperature to determine their potentiality for developing improved wheat cultivars. Introgression lines and their parent Chinese Spring were evaluated in a growth chamber at the seedling stage and in the field at the reproductive stage in two heat-stressed environments in Sudan. Optimum and late planting were used to ensure exposure of the plants to heat stress at the reproductive stage. The results revealed the impact of several Leymus chromosomes in improving wheat adaptation and tolerance to heat. Three lines possessed enhanced adaptation, whereas two showed high heat tolerance. Two addition lines showed a large number of kernels per spike, while one possessed high yield potential. Grain yield was correlated negatively with the heat susceptibility index, days to heading and maturity and positively with kernel number per spike and triphenyl tetrazolium chloride assay under late planting. The findings suggest that these genetic stocks could be used as a bridge to introduce the valuable Leymus traits into a superior wheat genetic background, thus helping maximize wheat yield in heat-stressed environments.     

Effect of <Emphasis Type="Italic">Acacia senegal</Emphasis> on growth and yield of groundnut,sesame and roselle in an agroforestry system in North Kordofan state,Sudan

Two field trials were conducted under rainfed conditions at El-Obeid Research Farm and Eldemokeya Forest Reserve, North Kordofan State during the growing seasons 2004/2005 and 2005/2006. The objective was to investigate the effect of Acacia senegal on the performance and yield of groundnut (Arachis hypogea), sesame (Sesamum indicum) and roselle (Hibiscus sabdariffa) in an agroforestry system. The two trials consisted of seven treatments: three represented the intercropping of groundnut, sesame and roselle with A. senegal, three without trees and one represented A. senegal alone. Data were recorded on soil physical and chemical properties, soil moisture content, crops fresh weight (kg/ha), dry weight (kg/ha), gum yield (g/picking) and crop yield (kg/ha). The trees at El-Obeid yielded no gum, whereas those at Eldemokeya were 15 years old and were tapped as part of the total harvest in the agroforestry plots. Land equivalent ratios (LER) and simple financial analyses of gross surpluses were used to evaluate the productivity of the different treatments. Fresh weight of groundnut, sesame and roselle was significantly different (P < .0.05) at both sites. Higher fresh weights were found under the intercropping system than the sole cropping system. This could be attributed to a shading effect that limits fruit production of the field crops more than vegetative growth. Dry weights were significantly greater for sesame and roselle in both sites, while that of groundnuts was not significantly different. In both sites, intercropping reduced the yield of sesame by 6 and 11% in the first season and 37 and 39% in the second season. The reduction in roselle yield was 19 and 28% in the first season and 15 and 8% in the second season. Yield reduction in groundnut was 35 and 17% in the first season and 35 and 11% in the second season. The combined analysis indicated that intercropping reduced groundnut yield by 26%, sesame by 21% and roselle by 20%. All the treatments gave LER of more than one—indicating the superiority of growing the field crops in intercropping over the sole cropping systems. The highest LER of (1.71) was obtained when roselle was intercropped with A. senegal, while the lowest LER (1.48) was obtained when groundnuts were intercropped with A. senegal. All the treatments gave positive net revenues, the highest being for intercropped roselle (438 SDG/ha). The intercropping of sesame gave the second highest net revenue (387 SDG/ha), while the sole roselle gave the lowest net revenue (97 SDG/ha).     

Effect of exogenous circulating anti-bPL antibodies on bovine placental lactogen measurements in foetal samples

Background

The involvement of placental lactogen (PL) in the regulation of foetal growth has been investigated in different species by in vivo immunomodulation techniques. However, when circulating antibodies are present together with the hormone, the procedure for hormonal measurement becomes considerably complex. The aim of this study was the immunoneutralization of bovine placental lactogen (bPL) concentrations in bovine foetal circulation by direct infusion of rabbit anti-bPL purified immunoglobulins (IgG) via a foetal catheter (in vivo study). The ability of a RIA based on guinea pig anti-bPL antiserum, for the measurement of bPL concentrations in samples containing exogenous rabbit anti-bPL immunoglobulins, was also analyzed in in vitro and in vivo conditions.

Methods

Six bovine foetuses were chronic cannulated on the aorta via the medial tarsal artery. Infusion of rabbit anti-bPL IgG was performed during late gestation. Pooled rabbit anti-bPL antisera had a maximal neutralization capacity of 25 μg bPL/mL of immunoglobulin. Interference of rabbit anti-bPL immunoglobulin with radioimmunoassay measurement using guinea pig anti-bPL as primary antibody was first evaluated in vitro. Polyclonal anti-bPL antibodies raised in rabbit were added in foetal sera to produce 100 samples with known antibodies titers (dilutions ranging from 1:2,500 till 1:1,280,000).

Result(s)

Assessment of the interference of rabbit anti-bPL antibody showed that bPL concentrations were significantly lower (P < 0.05) in samples added with dilutions of rabbit antiserum lower than 1:80,000 (one foetus) or 1:10,000 (four foetuses). It was also shown that the recovery of added bPL (12 ng/mL) was markedly reduced in those samples in which exogenous rabbit anti-bPL were added at dilutions lower than 1:20,000. Concentrations of foetal bPL were determined in samples from cannulated foetuses. In foetuses 1 and 6, bPL concentrations remained almost unchanged (<5 ng/mL) during the whole experimental period. In Foetus 3, bPL concentrations decreased immediately after IgG infusion and thereafter, they increased until parturition.

Conclusion(s)

The use of a bPL RIA using a guinea pig anti-bPL as primary antiserum allowed for the measurement of bPL concentrations in foetal plasma in presence of rabbit anti-bPL IgG into the foetal circulation. Long-term foetal catheterization allowed for the study of the influence of direct infusion of anti-bPL IgG on peripheral bPL concentrations in bovine foetuses.     

Genetic diversity of new maize hybrids based on SSR markers as compared with other molecular and biochemical markers

Genetic diversity of four new yellow single crosses, five new yellow three-way crosses, and five yellow inbred lines of maize (Zea mays L.) was studied using different molecular (SSR, ISSR, and RAPD) and biochemical markers (seed storage protein content). All markers were able to clearly separate the inbred lines in one cluster from the different types of hybrids. The correlation among the different types of molecular markers was moderately high according to the Mantel’s test (e.g. 0.67 between SSR and ISSR, 0.42 between SSR and RAPD, and 0.65 between ISSR and RAPD), indicating that the three techniques are efficient for evaluating genetic diversity in the maize genotypes. The correlation of biochemical markers (seed storage protein SDS-PAGE) with SSR, ISSR, and RAPD markers was 0.61, 0.74, and 0.48, respectively. It can be concluded that both molecular and biochemical markers are efficient to study the genetic diversity in maize. Among the different types of molecular markers, SSR is a more accurate marker-type because of its co-dominance and stability of results. It can also be said that biochemical and molecular markers are positively correlated and the correlation ranged between moderate to high. This could suggest using both marker types, separately or together, for genetic diversity studies in maize.     

Management of the root-knot nematode Meloidogyne incognita on tomato with combinations of different biocontrol organisms

The nematicidal effect of Pseudomonas fluorescens, Paecilomyces lilacinus, Pichia guilliermondii and Calothrix parietina singly or in combination was tested against root-knot nematode, Meloidogyne incognita. Treatments with P. fluorescens and P. lilacinus caused mortality of M. incognita as 45% and 30% of juveniles after 48 h of exposures, respectively compared to water control in vitro. Under greenhouse conditions, all treatments reduced the disease severity and enhanced plant growth compared to untreated control. Application of P. fluorescens, P. lilacinus and P. guilliermondii Moh 10 was more effective compared to C. parietina. There was a negative interaction between C. parietina and either P. lilacinus or P. guilliermondii. Fresh and dry weight of shoots and roots of plants were significantly reduced as a result of infection with M. incognita, however application of biocontrol agents singly or in mix recovered this reduction. Moreover, they enhanced the growth parameters compared with the control. Our results proved that application of different biocontrol agents (P. fluorescens, P. lilacinus and P. guilliermondii) not only has a lethal effect on nematode, but also enhances the plant growth, supplying many nutritional elements and induction the systemic resistance in plants. Presence of C. parietina as a soil inhabitant cyanobacterium could antagonize biocontrol agents leading to the reduction of their practical efficiency in soil.