Effects of heavy metals on aerobic denitrification by strain Pseudomonas stutzeri PCN-1

Effects of heavy metals on aerobic denitrification have been poorly understood compared with their impacts on anaerobic denitrification. This paper presented effects of four heavy metals (Cd(II), Cu(II), Ni(II), and Zn(II)) on aerobic denitrification by a novel aerobic denitrifying strain Pseudomonas stutzeri PCN-1. Results indicated that aerobic denitrifying activity decreased with increasing heavy metal concentrations due to their corresponding inhibition on the denitrifying gene expression characterized by a time lapse between the expression of the nosZ gene and that of the cnorB gene by PCN-1, which led to lower nitrate removal rate (1.67∼6.67 mg L⁻¹ h⁻¹), higher nitrite accumulation (47.3∼99.8 mg L⁻¹), and higher N₂O emission ratios (5∼283 mg L⁻¹/mg L⁻¹). Specially, promotion of the nosZ gene expression by increasing Cu(II) concentrations (0∼0.05 mg L⁻¹) was found, and the absence of Cu resulted in massive N₂O emission due to poor synthesis of N₂O reductase. The inhibition effect for both aerobic denitrifying activity and denitrifying gene expression was as follows from strongest to least: Cd(II) (0.5∼2.5 mg L⁻¹) > Cu(II) (0.5∼5 mg L⁻¹) > Ni(II) (2∼10 mg L⁻¹) > Zn(II) (25∼50 mg L⁻¹). Furthermore, sensitivity of denitrifying gene to heavy metals was similar in order of nosZ > nirS ≈ cnorB > napA. This study is of significance in understanding the potential application of aerobic denitrifying bacteria in practical wastewater treatment.

     

Effective biodegradation of 2,4,6-trinitrotoluene using a novel bacterial strain isolated from TNT-contaminated soil

In this environmental-sample based study, rapid microbial-mediated degradation of 2,4,6-trinitrotoluene (TNT) contaminated soils is demonstrated by a novel strain, Achromobacter spanius STE 11. Complete removal of 100 mg L⁻¹ TNT is achieved within only 20 h under aerobic conditions by the isolate. In this bio-conversion process, TNT is transformed to 2,4-dinitrotoluene (7 mg L⁻¹), 2,6-dinitrotoluene (3 mg L⁻¹), 4-aminodinitrotoluene (49 mg L⁻¹) and 2-aminodinitrotoluene (16 mg L⁻¹) as the key metabolites. A. spanius STE 11 has the ability to denitrate TNT in aerobic conditions as suggested by the dinitrotoluene and NO₃ productions during the growth period. Elemental analysis results indicate that 24.77 mg L⁻¹ nitrogen from TNT was accumulated in the cell biomass, showing that STE 11 can use TNT as its sole nitrogen source. TNT degradation was observed between pH 4.0–8.0 and 4–43 °C; however, the most efficient degradation was at pH 6.0–7.0 and 30 °C.     

Competition of As and other Group 15 elements for surface binding sites of an extremophilic Acidomyces acidophilus isolated from a historical tin mining site

An arsenic-resistant fungal strain, designated WKC-1, was isolated from a waste roaster pile in a historical tin mine in Cornwall, UK and successfully identified to be Acidomyces acidophilus using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS) proteomic-based biotyping approach. WKC-1 showed considerable resistance to As⁵⁺ and Sb⁵⁺ where the minimal inhibitory concentration (MIC) were 22500 and 100 mg L⁻¹, respectively, on Czapex-Dox Agar (CDA) medium; it was substantially more resistant to As⁵⁺ than the reference strains CBS 335.97 and CCF 4251. In a modified CDA medium containing 0.02 mg L⁻¹ phosphate, WKC-1 was able to remove 70.30% of As⁵⁺ (100 mg L⁻¹). Sorption experiment showed that the maximum capacity of As⁵⁺ uptake was 170.82 mg g⁻¹ dry biomass as predicted by the Langmuir model. The presence of Sb⁵⁺ reduced the As⁵⁺ uptake by nearly 40%. Based on the Fourier-transform infrared spectroscopy (FT-IR) analysis, we propose that Sb is competing with As for these sorption sites: OH, NH, CH, SO₃ and PO₄ on the fungal cell surface. To our knowledge, this is the first report on the impact of other Group 15 elements on the biosorption of As⁵⁺ in Acidomyces acidophilus.

     

Highly efficient regeneration and medicinal component determination of Phellodendron chinense Schneid

Phellodendron chinense Schneid is an important Chinese herb with berberine and phellodendrine in stems and leaves, but with little information available on in vitro culture of this species. Disinfection of explants in 75% alcohol for 45 s, sterilization in 0.1% HgCl₂ for 20 min, and submersion in 1.0 mol L⁻¹ gibberellin3 (GA₃) solution for 24 h was the optimal condition for seed germination. Murashige and Skoog’s (MS) medium supplemented with 2.0 mg L⁻¹ 6-benzylaminopurine (6-BA) in combination with 1.5 mg L⁻¹ 1-naphthylacetic acid (NAA) was optimal for callus induction. MS medium supplemented with 2.0 mg L⁻¹ 6-BA was the appropriate medium for induction of adventitious shoots, and 1/2MS medium supplemented with 2.0 mg L⁻¹ indole-3-butytric acid (IBA) and 0.5% active carbon was the optimal medium for root induction. The 15-d survival rate of regenerated plantlets after transplanting to basins containing perlite and peat moss (1:4) was greater than 80%, and the berberine and phellodendrine accumulation was lower in callus compared with regenerated plantlets. The establishment of highly efficient regeneration system provides technical support for genetic breeding of Phellodendron chinense Schneid.

     

An efficient callus-based in vitro regeneration protocol for Warburgia Ugandensis Sprague,an important medicinal plant in Africa

Warburgia ugandensis Sprague is a woody species in the family Canellaceae and an important source of medicines in Africa. Natural propagation of W. ugandensis is problematic due to its recalcitrant seeds and lack of an efficient in vitro regeneration system for this species. This study describes an efficient regeneration protocol. Petiole bases and shoot tips were used as explants. Callus tissue developed when the explants were cultured on Murashige and Skoog medium containing 30 g L⁻¹ sucrose and 7 g L⁻¹ agar (MS30 medium), supplemented with 1.0 mg L⁻¹ indole-3-butyric acid (IBA), 1.6 mg L⁻¹ 6-benzylaminopurine (BA), and 0.1 mg L⁻¹ thidiazuron (TDZ). Adventitious buds were efficiently induced from the callus when the MS30 medium was supplemented with 0.8 mg L⁻¹ BA and 0.2 mg L⁻¹ IBA. Root induction occurred within 7–10 d on half-strength MS30 medium supplemented with 0.8–1.0 mg L⁻¹ 1-napthalene acetic acid (NAA), 0.2 mg L⁻¹ IBA, and 0.03% (w/v) activated charcoal (AC). Roots were followed by root elongation on the same medium but lacking NAA and IBA. Approximately 50% of the plantlets cultured produced roots, while more than 80% of the plantlets survived and successfully grew to maturity.

     

Genetic homogeneity and high shoot proliferation in banana (Musa acuminata Colla) by altering medium thiamine level and sugar type

To enhance the multiplication rate in Musa acuminata Colla (banana; ‘Grand Nain’) organogenesis, higher amounts of thiamine along with different sugar types and concentrations were evaluated at the proliferation phase. Thiamine at 1, 10, 50, 100, and 200 mg L⁻¹ was compared with 0.1 mg L⁻¹ thiamine found in conventional Murashige and Skoog (MS) medium. Maximum proliferation of banana was induced with 100 mg L⁻¹ thiamine. Additionally, 15, 30, and 45 g L⁻¹ sucrose, glucose, fructose, and sorbitol combined with regular and optimal levels of thiamine were tested. Glucose at 30 g L⁻¹ most improved shoot proliferation alone and enhanced shoot proliferation further, when combined with 100 mg L⁻¹ thiamine, followed by sucrose and fructose, whereas sorbitol completely inhibited growth and caused tissue browning. All evaluated vegetative traits were significantly affected by sugar type and concentration, and thiamine levels, unlike the photosynthetic pigments. Moreover, genetic stability of the plants recovered from the enhanced protocol was confirmed by inter-simple sequence repeats (ISSR) and randomly amplified polymorphic DNA (RAPD) analysis. A total of 230 bands generated by both marker types were monomorphic for the randomly selected regenerated plants, compared with their mother plant. Thus, the proliferation medium supplemented with 30 g L⁻¹ glucose and 100 mg L⁻¹ thiamine could be recommended for banana organogenesis. Results herein are of great importance and helpful in enhancing the commercial in vitro propagation protocols of banana, without the need of increasing the number of subcultures, which can cause somaclonal variation.

     

Phytotoxic and genotoxic effects of silver nanoparticles exposure on germinating wheat seedlings

We investigated the effects of 1 and 10 mg L⁻¹ AgNPs on germinating Triticum aestivum L. seedlings. The exposure to 10 mg L⁻¹ AgNPs adversely affected the seedling growth and induced morphological modifications in root tip cells. TEM analysis suggests that the observed effects were due primarily to the release of Ag ions from AgNPs.     

Nanosilver inhibits nitrification and reduces ammonia‐oxidising bacterial but not archaeal amoA gene abundance in estuarine sediments

Silver nanoparticles (AgNPs) enter estuaries via wastewater treatment effluents, where they can inhibit microorganisms, because of their antimicrobial properties. Ammonia‐oxidising bacteria (AOB) and archaea (AOA) are involved in the first step of nitrification and are important to ecosystem function, especially where effluent discharge results in high nitrogen inputs. Here, we investigated the effect of a pulse addition of AgNPs on AOB and AOA ammonia monooxygenase (amoA) gene abundances and benthic nitrification potential rates (NPR) in low‐salinity and mesohaline estuarine sediments. Whilst exposure to 0.5 mg L^?1 AgNPs had no significant effect on amoA gene abundances or NPR, 50 mg L^?1 AgNPs significantly decreased AOB amoA gene abundance (up to 76% over 14 days), and significantly decreased NPR by 20‐fold in low‐salinity sediments and by twofold in mesohaline sediments, after one day. AgNP behaviour differed between sites, whereby greater aggregation occurred in mesohaline waters (possibly due to higher salinity), which may have reduced toxicity. In conclusion, AgNPs have the potential to reduce ammonia oxidation in estuarine sediments, particularly where AgNPs accumulate over time and reach high concentrations. This could lead to long‐term risks to nitrification, especially in polyhaline estuaries where ammonia‐oxidation is largely driven by AOB.     

Modification of light intensity influence essential oils content,composition and antioxidant activity of thyme,marjoram and oregano

Thymus vulgaris L. (thyme), Origanum majorana L. (marjoram), and Origanum vulgare L. (oregano) were used to determine whether light modification (plants grown under nets with 40% shaded index or in un-shaded open field) could improve the quantity and quality of essential oils (EOs) and antioxidant activity. The yield of EOs of thyme, marjoram, and oregano obtained after 120 min of hydrodistillation was 2.32, 1.51, and 0.27 mL/100 g of plant material, respectively. At the same time under shading conditions plants synthetized more EOs (2.57, 1.68, and 0.32 mL/100 g of plant material). GC/MS and GC/FID analyses were applied for essential oils determinations. The main components of the thyme essential oil are thymol (8.05–9.35%); γ-terpinene (3.49–4.04%); p-cymene (2.80–3.60%) and caryophyllene oxide (1.54–2.15%). Marjoram main components were terpinene 4-ol (7.44–7.63%), γ-terpinene (2.82–2.86%) and linalool (2.04–2.65%) while oregano essential oil consisted of the following components: caryophyllene oxide (3.1–1.93%); germacrene D (1.17–2.0%) and (E)-caryophyllene (1.48–1.1%). The essential oil from thyme grown under shading (EC₅₀ value after 20 min of incubation) have shown the highest antioxidant activity – 0.85 mg mL⁻¹ in comparison to marjoram and oregano (shaded plants EC₅₀ 19.97 mg mL⁻¹ and 7.02 mg mL⁻¹ and unshaded, control plants EC₅₀ 54.01 mg mL⁻¹ and 7.45 mg mL⁻¹, respectively). The medicinal plants are a good source of natural antioxidants with potential application in the food and pharmaceutical industries. For production practice, it can be recommended to grow medicinal plants in shading conditions to achieve optimal quality parameters.     

Acclimation of arsenic-resistant Fe(II)-oxidizing bacteria in aqueous environment

This study investigated the potential of the Fe(II)-oxidizing bacteria in removing arsenic in aqueous environment. The bacteria were isolated from the batch of tap water and rusty iron wires, and were acclimated to culture media amended with arsenic concentrations, gradually increasing from 100 μg L⁻¹ to 100 mg L⁻¹. Acclimated bacteria with enhanced arsenic tolerance were used to remove arsenic from the aqueous solution. These bacteria belonged to Pseudomonas species according to 16S rRNA gene sequences. Extracellular enzymes produced by these bacteria played important roles in microbial Fe(II) oxidization and Fe oxide precipitation. Moreover, these bacteria survived and propagated in high arsenic condition (100 mg L⁻¹ As). However, after As(III/V) acclimation, morphological characteristics of the bacteria showed some changes, e.g., shrinking of long bacillus. XRD (X-ray diffraction) patterns indicated that Fe oxide precipitations by Fe(II)-oxidizing bacteria in Fe-rich culture medium were poorly-crystallized ferrihydrites. Adsorption on the biogenic ferrihydrites greatly contributed to high arsenic removal efficiency of Fe(II)-oxidizing bacteria.     

Use of bioreactors for large-scale multiplication of sugarcane (Saccharum spp.), energy cane (Saccharum spp.), and related species

The objective of this study was to set up a plant micropropagation facility to mass propagate sugarcane, energy cane, and related clonally propagated species. An efficient methodology for micropropagation of energy cane and perennial grasses using temporary immersion bioreactors was developed. Several different methods of tissue culture initiation, multiplication, and rooting were evaluated for several varieties of sugarcane (Saccharum officinarum L.) and sugarcane-related species such as Erianthus spp., Miscanthus spp., and Sorghum spp. × sugarcane hybrids, all from a germplasm collection. Apical meristem cultures were initiated for all genotypes that were micropropagated, when liquid or semisolid Murashige and Skoog (MS) medium was used, which was supplemented with 0.1–0.2 mg L⁻¹ BAP, 0.1 mg L⁻¹ kinetin, 0–0.1 mg L⁻¹ NAA, and 0–0.2 μg L⁻¹ giberellic acid. These cultures produced shoots between 4 and 8 wk after initiation. Shoot regeneration from leaf rolls or immature inflorescences was observed as early as 4 wk after initiation. Shoot multiplication was successful for all genotypes cultured in MS medium with 0.2 mg L⁻¹ BAP and 0.1 mg L⁻¹ kinetin. Energy cane had a significantly higher combined multiplication rate when grown under four or five LED lamps than when grown under three LED lamps, or under fluorescent lights in a growth chamber. The addition of 2 mg L⁻¹ NAA produced faster and better rooting in all of the genotypes tested. Shoots produced well-developed roots after one cycle of 15–21 d in the bioreactors. The maximum number of plantlets produced per bioreactor was 1080. Plantlets developed a vigorous root system and were ready to be transplanted into the field after 2 mo. A protocol was standardized for different energy cane clones that were recommended for their biomass production and cell wall composition. Different tissues were used to speed up or facilitate tissue culture initiation. Visual assessment of micropropagated plants in the field did not show any off-types, based on gross morphological changes of plant morphology or disease reaction, compared to plants of the same genotype derived from a traditional propagation method (stem cuttings). This is the first report of energy cane and Miscanthus spp. micropropagation using the SETIS bioreactor.

     

Effect of molybdenum on secondary metabolic process of glycyrrhizic acid in Glycyrrhiza uralensis Fisch.

A pot experiment was conducted to investigate into effects of molybdenum (Mo) on the secondary metabolic process of glycyrrhizic acid (GA). One-year-old seedlings were grown in pots with washed vermiculite and sand. Hoagland nutrition solution was irrigated with four concentrations: 0, 0.52, 5.2 and 10.4 mg L⁻¹. The accumulations of GA and its biosynthetic precursors (β-amyrin and squalene) and then expression of the key synthase (β-amyrin synthase, β-AS) were studied on 35, 70 and 105 d. In the early stage, that was on the 35 and 70 d, the contents of squalene and GA, and the expression of β-AS gene under 0.52 and 5.2 mg L⁻¹ Mo treatments were significantly higher than that under 0 and 10.4 mg L⁻¹ Mo. There was a contrary result of β-amyrin. However, the content of squalene under 0 mg L⁻¹ Mo was the highest on 105 d. Thus, it suggested an appropriate concentration of Mo could promote the accumulation of GA, by affecting the biosynthetic process of GA at a certain time. Practically, the time and amount of application of Mo on Glycyrrhiza uralensis should be the noted.     

Influence of linear alkylbenzene sulfonate and ethanol on the degradation kinetics of domestic sewage in co-digestion with commercial laundry wastewater

The influence of ethanol on the degradation kinetics of linear alkyl benzene sulfonate (LAS) and organic matter was investigated using batch experiments with different initial LAS concentrations (8.3 mg L⁻¹ to 66.9 mg L⁻¹) and biomass immobilized on sand. Data were fitted with a substrate inhibition model. Concentrations of 2.4 mg LAS L⁻¹ and 18.9 mg LAS L⁻¹ (without and with ethanol) provided the maximum LAS utilization rate by the biomass (S_bm). For LAS degradation, ethanol addition favored a lower decrease in the specific substrate utilization rate (r_obs), even at the LAS concentration usually reported as inhibitory (> 14.4 mg L⁻¹). For organic matter degradation, r_obs was higher with ethanol. Higher biomass differentiation was observed at higher LAS concentrations. With ethanol, microbial selection occurred at LAS concentrations near S_bm. At higher LAS concentrations, the dominance and diversity values did not change significantly with ethanol, whereas without ethanol, their behaviors were irregular.

     

Synergistic effect of cytokinins and auxins enables mass clonal multiplication of drumstick tree (Moringa oleifera Lam.): a wonder

The synergistic effect of plant growth regulators on axillary bud proliferation for mass clonal multiplication of Moringa oleifera Lam. (vern. drumstick) has been assessed for the first time. Treatment of decoated seeds with 1% (w/v) Bavistin for 60 min, 0.33% (w/v) streptocycline for 30 min, and 0.1% (w/v) HgCl₂ for 3.5 min resulted in complete removal of the surface contaminants. Maximum seed germination (89.13%) was obtained on quarter-strength Murashige & Skoog (MS) medium. Culture of nodal segments on MS + 6-benzyladenine (BA) at 3 mg L⁻¹ resulted in multiple shoot proliferation with ~ 18 shoots per explant. All combinations of indole-3-acetic acid (IAA) + kinetin (Kn) resulted in elongated shoots, while only lower concentrations of BA (0.5 mg L⁻¹), along with IAA (0.5 to 2 mg L⁻¹), or Kn (0.5 to 5 mg L⁻¹), showed significant synergy in the shoot morphogenesis. In addition, the maximum (100%) rooting efficiency was attained on half-strength MS medium supplemented with different concentrations of IAA and indole-3-butyric acid (IBA). The rooted plants were successfully established in the greenhouse for acclimatization. Clonality of the raised plants was assessed using 15 random primers of Operon® technologies (OPT and OPF series), and eight primers resulted in significant amplification with distinct, identical, and reproducible bands that confirmed clonality of the micropropagated plants. The present study provides a comprehensive analysis of the synergistic effect of plant growth regulators (PGRs) on in vitro shoot regeneration and proliferation for clonal mass multiplication disease-free plantlets, which can be utilized to maximize the yield of healthy and genetically identical plants of drumstick tree, which is considered to be a miracle multipurpose tree.

     

Microbial nitrogen removal of ammonia wastewater in poly (butylenes succinate)-based constructed wetland: effect of dissolved oxygen

Constructed wetland (CW) is popular in wastewater treatment for its prominent advantage of low construction and operation cost. However, the nitrogen removal in conventional CW is usually limited by the low dissolved oxygen (DO) and insufficient electron donor. This paper investigated the nitrogen removal performance and mechanisms in the poly (butylenes succinate)-based CW (PBS-CW) while treating ammonia wastewater under different DO levels. The average DO contents in limited-aeration and full-aeration phases were 1.68 mg L⁻¹ and 5.71 mg L⁻¹, respectively. Results indicated that, with the ammonia nitrogen loading rate of 25 g N m⁻³ day⁻¹, total nitrogen removal ratios in the PBS-CW under the limited-aeration and full-aeration phases were 72% and 99%, respectively. Combined analyses revealed that simultaneous nitrification and denitrification (SND) via nitrite/nitrate were the main microbial nitrogen removal pathways in the aeration phase of the PBS-CW (> 89%). The microbial carrier of biodegradable material was believed to play a significant role in prompting SND performance while dealing with low C/N wastewater. Due to the coexistence of micro-anaerobic zone and carbon supply inside the coated biofilm, the high DO level in the PBS-CW increased the abundance of the nitrifying bacteria (amoA and nxrA), denitrifying bacteria (narG, nirK, nirS, and nosZ), and even anammox bacteria (anammox 16s rRNA). These features are beneficial to many microbial processes which require the simultaneous aerobic, anoxic, and anaerobic environment.

     

Soil and tree chemistry reflected the cumulative impact of acid deposition in Pinus banksiana and Populus tremuloides stands in the Athabasca oil sands region in western Canada

The spatial variability of soil chemistry and Ca/Al ratios of soil solution and fine roots were investigated in jack pine (Pinus banksiana) and trembling aspen (Populus tremuloides, aspen) stands to assess the impact of chronic acid deposition on boreal forest ecosystems in the Athabasca oil sands region (AOSR) in Alberta, Canada. Available SO₄²⁻ (as the sum of soluble and adsorbed SO₄²⁻) accumulated in the soil near tree boles of both species, reflecting the influence of canopy intercepted SO₄²⁻. In jack pine stands, pH and soluble base cation concentrations decreased towards tree boles due to increased SO₄²⁻ leaching; the reverse was found in aspen stands due to deposition of base cations leached from the canopy. As a result, Ca/Al ratios in the soluble fraction in soils near jack pine boles were 5–20 times lower than that near aspen boles. The Ca/Al ratio did not reach the critical limits of 1.0 for soil solution (ranged from 1.0 to 4.1) or 0.5 for fine roots (0.7–7.9) in the studied watersheds. However, Alⁿ⁺ concentrations in the soil solution ranged from 0.2 to 4.1 mg L⁻¹ in NE7 and from 0.1 to 8.5 mg L⁻¹ in SM8 that can inhibit the growth of white spruce (Picea glauca) seedlings that commonly succeed aspen in upland sites in the AOSR. We suggest that the spatial variation caused by tree canopies/stems will affect forest regeneration and the effect of acid deposition on forest succession in the AOSR should be further studied.     

Biodegradation of methyl parathion in the presence of goethite: The effect of Pseudomonas sp. Z1 adhesion

In this research, the influence of goethite on biodegradation kinetic of methyl parathion was investigated in the presence of Pseudomonas sp. Z1. Semipermeable membrane experiments were performed to demonstrate the role of adhesion of degrading bacteria to surface of goethite in biodegradation of methyl parathion. Sorption of methyl parathion and bacteria onto goethite particles were also measured to assess the distribution of methyl parathion and bacteria between water and goethite surface. The first-order degradation rate constant of methyl parathion in different concentrations of goethite was in the order of 0.1 g L⁻¹ > 0.01 g L⁻¹ > 0 g L⁻¹ > 1 g L⁻¹ > 20 g L⁻¹, suggesting the presence of low concentrations of goethite accelerated the biodegradation of methyl parathion and high concentrations of goethite inhibited this biodegradation process. According to the result of semipermeable membrane experiment, when no bacterial attachment occurred in the system, the promotive effect of 0.1 g L⁻¹ goethite for microbial degradation was disappeared and the inhibition effect of 20 g L⁻¹ goethite increased. The results clearly demonstrated that the adhesion of bacteria to goethite was beneficial to the biodegradation of methyl parathion. The information obtained is of fundamental significance for the understanding of microbial degradation of organic pollution in soil.     

Smoke-saturated Water and Karrikinolide Modulate Germination,Growth, Photosynthesis and Nutritional Values of Carrot (Daucus carota L.)

Plant-derived smoke is a positive regulator of seed germination and growth with regard to many plant species. Of the several compounds present in plant-derived smoke, karrikinolide or KAR₁ (3-methyl-2H-furo[2,3-c]pyran-2-one) is considered to be the major active growth-promoting compound. To test the efficacy of smoke-saturated water (SSW) and KAR₁ on carrot (Daucus carota L.), two separate pot experiments were simultaneously conducted in the same environmental conditions. SSW and KAR₁ treatments were applied to the plants in the form of aqueous solutions of variable concentrations. Prior to sowing, seeds were soaked in the solutions of SSW (25.8 µg L⁻¹_, 51.6 µg L⁻¹,103.2 µg L⁻¹ and 258.0 µg L⁻¹) and KAR₁ (0.015 µg L⁻¹, 0.150 µg L⁻¹, 1.501 µg L⁻¹ and 15.013 µg L⁻¹). Percent seed germination, vegetative growth, photosynthesis and nutritional values were the major parameters through which the plant response to the applied treatments was investigated. The results obtained indicated a significant improvement in all the plant attributes studied. In general, SSW (51.6 µg L⁻¹) and KAR₁ (1.501 µg L⁻¹) proved optimum treatments for most the parameters. As compared to the control, 51.6 µg L⁻¹ of SSW and 1.501 µg L⁻¹ of KAR₁ increased the percent seed germination by 58.0% and 54.4%, respectively. Over the control, the values of plant height and net photosynthetic rate were enhanced by 33.9% and 40.9%, respectively, due to 51.6 µg L⁻¹ of SSW, while the values of these parameters were increased by 25.2% and 34.0%, respectively, due to 1.501 µg L⁻¹ of KAR₁. In comparison with the control, treatment 51.6 µg L⁻¹ of SSW increased the contents of β-carotene and ascorbic acid by 32.7% and 37.9%, respectively, while the treatment 1.501 µg L⁻¹ M of KAR₁ enhanced these contents by 42.0% and 48.4%, respectively. This study provides an insight into an affordable and feasible method of crop improvement.

     

Somatic embryogenesis and plant regeneration in two wild cotton species belong to G genome

The present work describes the plant regeneration via somatic embryogenesis in two wild cotton species belonging to G genome: Gossypium nelsonii Fryx and Gossypium australe F Muell. The role of plant hormones and carbohydrates was also evaluated for somatic embryogenesis and somatic embryo development. Normal plants were obtained from G. nelsonii Fryx; abnormal plants and somatic embryos were obtained from G. australe F Muell. The best medium for callus induction for these G genome wild cotton species was MSB₅ supplemented with 0.1 mg L⁻¹ KT and 0.1 mg L⁻¹ 2,4-D. For embryogenic callus proliferation, the best medium used was MSB₅ supplemented with 0.2 mg L⁻¹ KT and 0.5 mg L⁻¹ IBA. The medium MSB₅ supplemented with 0.15 mg L⁻¹ KT and 0.5 mg L⁻¹ NAA was used successfully for root initiation and plant growth. In addition, adding CuSO₄ and AgNO₃ in the callus-inducing and proliferation medium resulted in a number of somatic embryos. Glucose and maltose, the carbon sources in somatic culture, were used for callus induction, but maltose worked even better than glucose for proliferation of embryogenic callus and development of somatic embryos.     

Somatic embryogenesis to overcome low seed viability and conserve wild banana (Ensete superbum (Roxb.) Cheesman)

The seed viability, ex vitro germination, and percentage of in vitro zygotic embryo germination were found to be very low in Ensete superbum (Roxb.) Cheesman. Only 33.33% of seeds were viable, and the ex vitro germination percentage was only 5%, while the percentage of in vitro zygotic embryo germination was 33%. Somatic embryogenesis experiments produced competent callus on Murashige and Skoog (MS) medium supplemented with 2.5 mg L⁻¹ 2,4-D and 3 mg L⁻¹ BAP from inflorescence explants. The embryogenic callus produced the maximum number of somatic embryos on MS basal medium kept in a dark chamber for 15 wk. Half-strength MS medium supplemented with 500 mg L⁻¹ glutamine was optimal for somatic embryo germination and development of plantlets. Regenerated plants had 80 to 90% survival rate. Therefore, somatic embryogenesis can be considered as an efficient method to overcome a drastic reduction in population and to achieve germplasm conservation.