Effect of Sodium Azide on Quantitative and Qualitative Stem Traits in the M2 Generation of Ethiopian Sesame (Sesamum indicum L.) Genotypes.

The emerging oilseed crop Sesamum indicum, also known as the queen of oilseeds, is being grown globally for its oil content for medicinal and nutritional values. One of the key challenges of sesame cultivation is its low productivity. In the present study, sodium azide (NaN3) was used as a chemical mutagen. The aim of this study was to examine the effect of NaN3 on quantitative and qualitative stem traits in the M2 generation of Ethiopian sesame (Sesamum indicum L.) genotypes. Seeds of fourteen sesame genotypes were used in this study and germinated and grown under greenhouse conditions. Different qualitative and quantitative data were collected and analyzed. Traits such as plant height, ground distance to first distance, and internode length were significantly affected by NaN3 treatment. The highest plant height was recorded in the control on Humera 1 and Baha Necho genotypes, while the lowest was observed on Setit 2 and Hirhir treated with the chemical. The highest ground distance to the first branch was observed in Gumero, while the least ground distance was recorded in Setit 1 in the treated and control genotypes, respectively. The best internode length was recorded on Setit 2 and ADI in the control, while the lowest internode length was observed in Setit 1 genotype treated with sodium azide. Genotypes such as ACC44, ADI, Baha Necho, Borkena, Gonder 1, and Setit 1 treated with NaN3 have showed glabrous type of stem hairiness. All the fourteen genotypes (both treated and control) were clustered into four groups. In conclusion, we observed a highly significant variation among the genotypes due the effect of the chemical and genotypes themselves. Hence, this report would create more genetic diversity for further sesame genetic research improvements.

Impact of different amendments on biochemical responses of sesame (Sesamum indicum L.) plants grown in lead-cadmium contaminated soil.

Soil co-contamination with lead (Pb) and cadmium (Cd) is a tenacious risk to crop production globally. The current experiment observed the roles of amendments [biochar (BC), slag (SL), and ferrous manganese ore (FMO)] for enhancing Pb and Cd tolerance in sesame (Sesamum indicum L.). Our results revealed that application of amendments significantly enhanced the nutrient level of sesame seedlings developed under extreme Pb and Cd conditions. The higher Pb and Cd-tolerance in sesame encouraged by amendments might be credited to its capability to restrict Pb and Cd uptake and decreased oxidative damage induced by Pb and Cd that is also demonstrated by lesser production of hydrogen peroxide (H2O2), malondialdehyde (MDA), and reduced electrolyte leakage (EL) in plant biomass. The added amendments relieved Pb and Cd toxicity and improved photosynthetic pigments, soluble protein, and proline content. Not only this amendments also decreased the antioxidant bulk, such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in sesame plants compared to control when exposed to Pb and Cd. Moreover, the added amendments = down-regulated the genes expression which regulate the SOD, POD, and CAT activity in sesame under Pb and Cd-stress. Furthermore, supplementation of amendments to the soil, reduced the bio accessibility (SBET), leachability (TCLP), and mobility (CaCl2) of Pb and Cd. Collectively, our findings conclude that the application of amendments enhanced sesame tolerance to Pb and Cd stress by restricting Pb and Cd accumulation, maintained photosynthetic presentation and dropped oxidative loss through enhanced antioxidant system, thus signifying amendments as an operational stress regulators in modifying Pb and Cd-toxicity that is highly important economically in all crops including sesame.

Role of two-sided crosstalk between NO and H2S on improvement of mineral homeostasis and antioxidative defense in Sesamum indicum under lead stress.

H2S and NO are two important gasotransmitters that modulate stress responses in plants. There are the contradictory data on crosstalk between NO and H2S in the studies. Hence, in the present study, the role of interplay between NO and H2S was assessed on the Pb tolerance of Sesamum indicum using pharmacological and biochemical approaches. Results revealed that Pb stress reduced the plant growth and the content of photosynthetic pigments and Fv/Fm ratio, increased the lipid peroxidation and the H2O2 content, elevated the endogenous contents of nitric oxide (NO), H2S and enhanced the activities of antioxidant enzymes (except APX). Additionally, concentrations of most mineral ions (K, P, Mg, Fe, Mn and Zn) in both shoots and roots decreased. Pb accumulation in roots was more than it in shoots. Both sodium hydrosulfide (NaHS as a donor of H2S) and sodium nitroprusside (SNP as an NO donor) improved the plant growth, the chlorophyll and carotenoid contents and PSII efficiency, reduced oxidative damage, increased the activities of antioxidant enzymes and reduced the proline content in Pb-stressed plants. Furthermore, both NaHS and SNP significantly restricted the uptake and translocation of Pb, thereby minimizing antagonistic effects of Pb on essential mineral contents in sesame plants. NaHS increased the NO generation and many NaHS-induced responses were completely reversed by cPTIO, as the specific NO scavenger. Applying SNP also enhanced H2S release levels in roots of Pb-stressed plants and only some NO-driven effects were partially weakened by hypotuarine (HT), as the scavenger of H2S.These findings proposed for the first time that two-sided interplay between H2S and NO might confer an increased tolerance to Pb stress via activating the antioxidant systems, reducing the uptake and translocation of Pb, and harmonizing the balance of mineral nutrient.

Characterization of phytotoxin producing Alternaria species isolatedfrom sesame leavesand their toxicity.

Sesame (Sesamum indicum L.), is an important oilseed crop in the tropics and subtropics, referred as "Queen of Oilseeds" owing to its high cooking quality and medicinal value. Sesame production, particularly in India, has been declining since last decade and 'Leaf blight' caused by Alternaria spp. is reported to cause yield loss up to 30-40%. Here, we investigated the fungal toxin produced by Alternaria and its pathogenicity. A total of 164 Alternaria strainswere isolated on potato dextrose agar media from the infected sesame leaves showing circular concentric rings with dark brown spots symptoms. All the isolates were screened for cultural and morphological characters. Colour of the fungus was grey to dark brown, formed smooth, raised, fluffy, and regular to irregular margins. Among 164 isolates, 43 isolates were moderately growing and 121 were fast in growth. The DNA of the isolate was amplified with ITS primers and sequence of BLAST results confirmed seven different species of Alternaria of NCBI database. Further, toxigenic potentiality of the isolates was tested with dilutions of culture filtrate (1:1 to 1:5) on sesame leaves. Among 164 isolates, 23 showed toxigenicity, varied from highly toxigenic to least toxigenic. Pathogenicity of the isolates showed that they were highly virulent to less virulent when tested by the detached leaf method. Based on the toxigenicity, the toxin was partially purified and brown coloured paste was recovered. Chemistry of the toxin was confirmed based on the IR, UV, NMR and mass spectra analyses, and it resembled the structure of alternariol mono methyl ether and altenuene which are mycotoxins in nature. Further, bioassay of toxin was carried out at different concentrations (50 to 2000 ppm) on seeds and seedlings of sesame. Maximum inhibition of seed germination of 81.1% was observed at 2000 ppm and the least was 6.67% at 50 ppm. With the increase in the concentration of toxin, the manifestation of the symptom was conspicuous and quick such as marginal, veinal necrosis, drooping and yellowing with lesion formation. From the present study, it is found that the species of Alternaria are responsible for the cause of blight disease symptoms and the toxicity of toxin produced by the pathogen was very high. The Alternaria toxin could inhibit the growth of the plant as well as seed germination rate.

A new high-frequency Agrobacterium-mediated transformation technique for Sesamum indicum L. using de-embryonated cotyledon as explant.

In spite of the economic importance of sesame (Sesamum indicum L.) and the recent availability of its genome sequence, a high-frequency transformation protocol is still not available. The only two existing Agrobacterium-mediated transformation protocols that are available have poor transformation efficiencies of less than 2%. In the present study, we report a high-frequency, simple, and reproducible transformation protocol for sesame. Transformation was done using de-embryonated cotyledons via somatic embryogenic stages. All the critical parameters of transformation, like incubation period of explants in pre-regeneration medium prior to infection by Agrobacterium tumefaciens, cocultivation period, concentrations of acetosyringone in cocultivation medium, kanamycin concentration, and concentration of plant hormones, including 6-benzylaminopurine, have been optimized. This protocol is superior to the two existing protocols in its high regeneration and transformation efficiencies. The transformed sesame lines have been tested by PCR, RT-PCR for neomycin phosphotransferase II gene expression, and ß-glucuronidase (GUS) assay. The regeneration frequency and transformation efficiency are 57.33 and 42.66%, respectively. T0 and T1 generation transgenic plants were analyzed, and several T1 plants homozygous for the transgenes were obtained.

[Induction of cytomixis affects microsporogenesis in Sesamum indicum L. (Pedaliaceae)].

Cytomixis was recorded during microsporogenesis in sesame (Sesamum indicum L.), a member of the family Pedaliaceae. The phenomenon of cytomixis was observed at various stages of meiosis in 0.5% Sodium azide (SA) treated populations of Sesamum indicum L. Cytomixis was observed to occur through various methods, i.e., by forming cytoplasmic channels and direct fusion of pollen mother cells (PMCs), the former was more frequent than the latter. The migration of nuclear content involved all the chromatin/chromosomes or part of it from donor to recipient cell/cells. Some completely empty meiocytes were also observed. Stickiness, precocious movement, laggards, unorientation and micronuclei were observed in almost all the sets treated with various doses of SA. Increase in the doses of SA had a positive effect on the percentage of PMCs showing cytomixis and chromosomal abnormalities. The impact ofcytomixis on meiotic behaviour, reduced pollen viability and heterogeneous sized pollen grains were observed.

Accumulation of Cu, Zn, Pb, and Cd in edible parts of four commonly grown crops in two contaminated soils.

Soil heavy metal pollution resulting from human activities is causing major concern due to its potential risk. In this study, four crop species with different cultivars were planted in 2 levels (heavily and slightly) of heavy metal contaminated soils, and the accumulation of Cu, Zn, Pb, and Cd in the edible parts of the crops were investigated. Metal concentrations in sesame seeds grown in both soils exceeded both the Chinese Food Hygiene Standard (CFHS) and Codex Alimentarius Commission Standard (CACS), while the metal concentrations in all pepper cultivars in the slightly contaminated soil were below the CFHS and CACS. Other crops were generally in between in both soils. Among the tested crops, the order of soil-plant transfer factor (TF) was: sesame > green soybean > cowpea > pepper. Additionally, old fruit of cowpea contained larger amounts of metals than young fruit. It suggests that sesame should not be planted in the metal contaminated area, while pepper cultivar "Chaobianjiao No.1" may be an alternative to be grown in the slightly contaminated soil. There were differences in individual human susceptibilities to metals. Therefore, a comprehensive risk assessment should consider the frequency, amount and species consumed by human besides metal concentrations in crops.

Effect of growing Sesamum indicum L. on enhanced dissipation of lindane (1, 2, 3, 4, 5, 6-hexachlorocyclohexane) from soil.

The effect of growing Sesamum indicum L. on the dissipation of lindane (gamma-HCH) was studied in spiked soil. For this, S. indicum was grown with four different concentrations of lindane (5, 10, 15, and 20 microg g(-1)). Plant growth, yield, photosynthetic pigments, soluble protein, microbial biomass carbon, lindane uptake, residual lindane concentration in soil and percentage dissipation of lindane from soil were analyzed at 25, 90, and 124 d. The accumulation of lindane in test plants was linearly related to the soil concentration (r2 = 0.897-0.979). At maturity, the accumulation of lindane in S. indicum grown with four spiked concentrations reached up to 7.98, 13.72, 23.71, and 33.29 microg g(-1) dry matter, respectively. There was a marked difference in the dissipation of lindane in vegetated and non-vegetated soils (p < 0.01). After final harvesting, the residual lindane concentrations in four spiked concentrations were reduced by 77.56, 70.12, 62.51, and 58.7%, respectively. Agronomic practice for the onsite application of this species is discussed. Based on the present study, it was calculated that S. indicum could accumulate 2237-2611 mg lindane per acre after 124 d cultivation. S. indicum could thus be used for the phytoremediation of lindane contaminated soil.

Biological indicators capable of assessing thermal treatment efficiency of hydrocarbon mixture-contaminated soil.

In China, there are many special sites for recycling and washing the used drums, which release a variety of C5-C40 hydrocarbon mixture into the soil around the site. The remediation of these contaminated sites by thermal treatment is adopted ubiquitously and needs to be assessed. Here we report the feasibility of biological indicators applied to assess thermal treatment efficiency in such contaminated soil. A series of biological indicators, including seed germination index (SGI), root elongation index (REI), plant growth height, biomass, carbon dioxide evolved (CDE), soil respiration inhibition (SRI) and soil enzymatic activities, were employed to monitor or assess hydrocarbon mixture removal in thermal treated soil. The results showed that residual hydrocarbon mixture content correlated strongly negatively with SGI for sesamum (Sesamum indicum L.), plant height, and biomass for ryegrass (Lolium perenne L.) in the concentration ranges of 0-3990, 0-3170 and 0-2910 mg kg(-1), respectively. In contrast, REI for sesamum was positively correlated with residual hydrocarbon mixture content from 0 to 1860 mg kg(-1). In addition, both CDE and SRI demonstrated that 600 mg kg(-1) of residual hydrocarbon mixture content caused the highest amount of soil carbon dioxide emission and inhabitation of soil respiration. The results of soil enzymes indicated that 1000 mg kg(-1) of residual hydrocarbon mixture content was the threshold value of stimulating or inhibiting the activities of phosphatase and catalase, or completely destroying the activities of dehydrogenase, invertase, and urease. In conclusion, these biological indicators can be used as a meaningful complementation for traditional chemical content measurement in evaluating the environmental risk of the contaminated sites before and after thermal treatment.

Seed-expressed casein kinase I acts as a positive regulator of the SeFAD2 promoter via phosphorylation of the SebHLH transcription factor.

Microsomal oleic acid desaturase (FAD2) catalyzes the first committed step of the biosynthesis of polyunsaturated fatty acids via extra-plastidial desaturation of oleic acid to linoleic acid. In the regulatory mechanism controlling seed-specific SeFAD2 expression, trans-activation of the seed-specific SeFAD2 promoter is mediated by the SebHLH transcription factor (Kim et al. in Plant Mol Biol 64:453-466, 2007). In this study, a protein interacting with SebHLH was isolated from yeast two-hybrid analysis. The protein shares approximately 80% sequence identity with other putative casein kinases and was named SeCKI (Sesame Casein Kinase I). SeCKI transcripts were predominantly expressed in developing sesame seeds and were induced approximately threefold by exogenous application of ABA. eGFP:SeCKI fusion protein was localized to the nucleus. The SeCKI protein specifically bound to SebHLH. The SeCKI protein was autophosphorylated in a calcium-independent manner and transphosphorylated the SebHLH protein. Both the SebHLH and the SeCKI genes or both the SebHLH and mutated SemCKI (K182G) genes, under the control of CaMV 35S promoter, and the GUS reporter gene driven by SeFAD2 promoter containing E- and G-Box motifs were co-expressed in developing sesame seeds. This co-expression revealed that SeCKI enhanced the SebHLH-mediated transactivation of the SeFAD2 gene promoter via phosphorylation of the SebHLH transcription factor.

Seed-specific expression of sesame microsomal oleic acid desaturase is controlled by combinatorial properties between negative cis-regulatory elements in the SeFAD2 promoter and enhancers in the 5'-UTR intron.

The regulation of genes involved in primary lipid metabolism in plants is much less well understood than that in many other pathways in plant biology. In the investigation reported here, we have characterized transcriptional regulatory mechanisms controlling seed-specific FAD2 expression in sesame (Sesamum indicum). FAD2 codes for extra-plastidial FAD2 desaturase, which catalyzes the conversion of oleic acid to linoleic acid. Promoter analysis of the sesame FAD2 gene (SeFAD2) using the beta-glucuronidase (GUS) reporter system demonstrated that the - 660 to - 180 promoter region functions as a negative cis-element in the seed-specific expression of the SeFAD2 gene. Sesame and Arabidopsis FAD2 genes harbor one large intron within their 5'-untranslated region. These introns conferred up to 100-fold enhancement of GUS expression in transgenic Arabidopsis tissues as compared with intron-less controls. Prerequisite cis-elements for the SeFAD2 intron-mediated enhancement of gene expression and the promoter-like activity of SeFAD2 intron were identified. SeFAD2 transcripts were induced by abscisic acid (ABA) in developing sesame seeds, and the - 660 to - 548 and - 179 to - 53 regions in the SeFAD2 promoter were implicated in ABA-responsive signaling. Theses observations indicate that an intron-mediated regulatory mechanism is involved in controlling not only the seed-specific expression of the SeFAD2 gene but also the expression of plant FAD2 genes, which are essential for the synthesis of polyunsaturated fatty acids.

Identification of phytotoxic substances from early growth of barnyard grass (Echinochloa crusgalli) root exudates.

Barnyard grass is a problematic weed worldwide. It competes with crops and causes reduction in crop yields. In this study, barnyard grass suppressed rice emergence, and the degree of rice inhibition was proportional to the density of barnyard grass. Root exudates of barnyard grass reduced germination and growth of lettuce, rice, and monochoria. Fifteen compounds potentially involved in the phytotoxic activities of barnyard grass were isolated and identified, including phenolics, long-chain fatty acids, lactones, diethyl phthalate, acenaphthene, and derivatives of phthalic acids, benzoic acid, and decane. Quantities of diethyl phthalate, decanoic acid, myristic acid, stearic acid, 7,8-dihydro-5,6-dehydrokavain, and 7,8-dihydrokavain were 2.7, 11.1, 19.6, 35.5, 10.3, and 15.5 microg/ml of barnyard grass root exudates, respectively. The two lactones exhibited the greatest inhibition, followed by the phenolics and the derivatives of phthalic acids. Fatty acids had stronger suppression than diethyl phthalate and ethyl ester-4-ethoxy-benzoic acid. The acenaphthene and decane derivatives were the least phytotoxic. The phytotoxins released by barnyard grass roots showed strong inhibition on growth of broadleaf indicator plants and paddy weeds, but were less effective on barnyard grass itself and rice. Our study revealed that in addition to competition, barnyard grass also interferes with rice and other plants in its surroundings by chemical means.