A method for screening copper-tolerant rice (Oryza sativa L.) cultivars based on hydroponic experiments and cluster analysis

A method for screening copper (Cu)-tolerant rice cultivars was studied by combining hydroponic experiments and cluster analysis, and the classification of cultivars in Cu stress tolerance was done. In the first hydroponic experiment, seedlings of Jiahe991 and Xiushui114 were planted in nutrient solution with different Cu²⁺ concentrations from 10 to 1800 μg/L. Results indicated that the toxic threshold of Cu concentration in solution ranged from 900 to 1200 μg/L, since SPAD (Soil and Plant Analyzer Development, SPAD-502, a portable chlorophyll meter, Minolta Camera Co. Ltd., Japan) values of leaves and seedlings biomass of the treatments with ≥900 and/or 1200 µg/L were significantly lower than the control. The second experiment was conducted with 16 local rice cultivars under three Cu treatments (10, 1000, and 1500 μg/L). The 16 cultivars were well classified into tolerant, normal, and sensitive groups as a result of cluster analysis based on the relative SPAD (Soil and Plant Analyzer Development, SPAD-502, a portable chlorophyll meter, Minolta Camera Co. Ltd., Japan) value, shoot and root dry weights, root length and root dehydrogenase activity, and oxidizing capacity and shoot Cu concentration. Xiushui123, Xiushui134, Jiahe991, and Xianghu301 belonged to the tolerant group; Xiushui137 belonged to the sensitive group. The cluster analysis based on hydroponic experiments is an effective method for identifying rice cultivars that are tolerant to Cu stress. In addition, four cultivars (Xiushui123, Xiushui134, Jiahe991, and Xianghu301) are recommended in local practice.     

Comparison of gamma-aminobutyric acid production in Thai rice grains

Gamma-aminobutyric acid (GABA) has many pharmacological functions including being a major inhibitory neurotransmitter. Two comparative methods for GABA production in rice grains as main food source in Thailand were investigated in this study. Fermentation and germination method were separately carried out using seven selected local grain cultivars in northern Thailand. Red yeast rice, obtained from the fermentation method, gave the higher GABA concentration than the germinated rice produced from the germination method in most rice cultivars. The highest GABA concentration was 28.37 mg/g at 3 weeks fermentation time of glutinous rice, O. sativa L. cv. Sanpatong 1 cultivars, while germinated rice from glutinous rice; O. sativa L. cv. Korkor6 (RD6) cultivars contained the highest GABA concentration of 3.86 mg/g. These results provide information for the basis of an appropriate method for GABA production. The fermentation produced higher GABA concentration but required longer production period and red yeast rice was obtained as product. On the other hand, the germination method yielded rice grains with lower GABA but in more suitable form for consumption. Both methods are considered to be economical and efficient methods to increase GABA in rice grains, providing alternative products with higher nutritional values.     

Genetic characterization of chayote [<Emphasis Type="Italic">Sechium edule</Emphasis> (Jacq.) Swartz.] landraces of North Eastern Hills of India and conservation measure

Chayote or chow–chow is an underutilized cucurbit vegetable crop, widely cultivated by farmers in the backyards and Jhum lands for its tender leaves, fruits and tuberous root. In order to initiate crop improvement program in this crop, the present study was undertaken to assess the genetic variations in the 74 chow–chow landraces collected from the North Eastern Hill region of India. Wide variations for fruit colors, fruit length (6.5–21.5 cm), fruit width (4.2–10.7 cm), fruit weight (60–560 g), vitamin-C (2.6–13.8 mg/100 g), reducing sugar (0.18–2.77%), total sugar (1.09–2.94%) and phenol content (0.17–3.85 mg/100 g FW) were recorded among the landraces. All the landraces were also characterized using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers. In RAPD analyses, out of 28 primers a total of 198 reproducible amplicons were formed at an average of 7.01 per primer and an overall polymorphism of 88.38%. Eight fragments were specific to landraces with light green fruits. Four fragments were observed to be specific to RCSC-22 (dark green fruits) and another four specific to a RCSC-30 (pale yellow fruits). Out of 30 ISSR, only 5 primers generated a total of 32 reproducible amplicons with an average of 6.4 per primer and overall polymorphism of 62.5%. The pair wise similarity coefficient values ranged from 0.55 to 0.96. The grouping of landraces in cluster analysis was found to be independent of their respective geographic locations. The cuttings of suckers and shoot top (2 months old) treated with indole-3-butyric acid (200 mg l^?1) provide an alternative for the conservation of the diverse genetic materials to the researchers.     

A preliminary survey on the occurrence of mycotoxigenic fungi and mycotoxins contaminating red rice at consumer level in Selangor, Malaysia

Red rice is a fermented product of Monascus spp. It is widely consumed by Malaysian Chinese who believe in its pharmacological properties. The traditional method of red rice preparation disregards safety regulation and renders red rice susceptible to fungal infestation and mycotoxin contamination. A preliminary study was undertaken aiming to determine the occurrence of mycotoxigenic fungi and mycotoxins contamination on red rice at consumer level in Selangor, Malaysia. Fifty red rice samples were obtained and subjected to fungal isolation, enumeration, and identification. Citrinin, aflatoxin, and ochratoxin-A were quantitated by ELISA based on the presence of predominant causal fungi. Fungal loads of 1.4?×?10⁴ to 2.1?×?10⁶?CFU/g exceeded Malaysian limits. Monascus spp. as starter fungi were present in 50 samples (100 %), followed by Penicillium chrysogenum (62 %), Aspergillus niger (54 %), and Aspergillus flavus (44 %). Citrinin was present in 100 % samples (0.23–20.65 mg/kg), aflatoxin in 92 % samples (0.61–77.33 μg/kg) and Ochratoxin-A in 100 % samples (0.23–2.48 μg/kg); 100 % citrinin and 76.09 % aflatoxin exceeded Malaysian limits. The presence of mycotoxigenic fungi served as an indicator of mycotoxins contamination and might imply improper production, handling, transportation, and storage of red rice. Further confirmatory analysis (e.g., HPLC) is required to verify the mycotoxins level in red rice samples and to validate the safety status of red rice.     

Redox metabolic and molecular parameters for screening drought tolerant indigenous aromatic rice cultivars

The present work makes an effort to assess and standardize some redox metabolic and molecular parameters for screening drought tolerant indigenous aromatic rice cultivars of West Bengal, India. PEG-induced dehydration stress during early germination caused disruption of redox-homeostasis and oxidative damage in four IARVs (Jamainadu, Tulaipanji, Sitabhog and Badshabhog) by enhancing the accumulation of pro-oxidants [assessed in terms of oxidation of 2′,7′-dichlorofluorescindiacetate (DCFDA), accumulation of \({\text{O}}_{2}^{ \cdot - }\) and H₂O₂ and in situ staining of reactive oxygen species (ROS) in germinating tissue], significant reduction of antioxidative defence (total antioxidant and radical scavenging capacity, total thiol content and activities of antioxidative defence enzymes) and aggravating protein oxidation and lipid peroxidation (assessed in terms of free carbonyl content and accumulation of thiobarbituric acid reactive substances). When compared between the indigenous aromatic rice cultivars, a clear trend in differential redox regulatory properties in which ROS-antioxidant interaction acts at metabolic interface for redox homeostasis was observed in the order Badshabhog > Tulaipanji > Sitabhog > Jamainadu. Moreover, when the efficacy of ascorbate–glutathione cycle for scavenging H₂O₂ generated during dehydration stress was assessed and compared between the landraces exposed to PEG-induced dehydration stress in germinating tissue, it also exhibited almost the same trend with the landrace Tulaipanji and Badsabhog exhibiting maximum and Jamainadu the minimum efficiencies of the redox cycle. The indigenous aromatic rice cultivars Tulaipanji and Badsabhog resist dehydration stress better than the other two landraces due to its early preparedness to combat oxidative stress by up-regulating expression of genes of some enzymes of ascorbate–glutathione cycle along with some other antioxidative enzymes. A model of redox homeostasis in which ROS-antioxidant (ascorbate–glutathione system) acts at metabolic interface for up-regulation of antioxidative gene expression necessary for differential drought stress tolerance among the indigenous aromatic rice varieties is suggested.     

Boron application through seed coating improves the water relations, panicle fertility, kernel yield, and biofortification of fine grain aromatic rice

This study was conducted to evaluate the influence of boron (B) application through seed coating on leaf elongation, tillering, water relations, panicle sterility, kernel yield, and grain biofortification of fine grain aromatic rice. Boron was applied as seed coating at 1.0, 1.5, 2.0, 2.5, and 3.0 g B kg^?1 seed in two rice cultivars Super Basmati and Shaheen Basmati. Boron seed coating significantly affected the leaf elongation, water relations, panicle fertility, kernel yield and grain biofortification in both rice cultivars. However, seed coating with 1.0–2.0 g B kg^?1 seed was effective in improving the leaf emergence and elongation, and tillering. Whereas water relations (water and osmotic potential) were improved by all B seed coating treatments, but pressure potential was only improved from seed coating with 2.0–3.0 g B kg^?1 seed. Kernel yield was improved by all B seed coating treatments; however seed coating with 2 g B kg^?1 seed was the most effective treatment in this regard. Increase in kernel yield, by B seed coating, was attributed to decrease in panicle sterility. Leaf and kernel B contents were increased with increase in B concentration in seed coating.     

Genetic diversity among cultivars, landraces and wild relatives of rice as revealed by microsatellite markers

Genetic diversity among 35 rice accessions, which included 19 landraces, 9 cultivars and 7 wild relatives, was investigated by using microsatellite (SSR) markers distributed across the rice genome. The mean number of alleles per locus was 4.86, showing 95.2% polymorphism and an average polymorphism information content of 0.707. Cluster analysis based on microsatellite allelic diversity clearly demarcated the landraces, cultivars and wild relatives into different groups. The allelic richness computed for the clusters indicated that genetic diversity was the highest among wild relatives (0.436), followed by landraces (0.356), and the lowest for cultivars. Allelic variability among the SSR markers was high enough to categorize cultivars, landraces and wild relatives of the rice germplasm, and to catalogue the genetic variability observed for future use. The results also suggested the necessity to introgress genes from landraces and wild relatives into cultivars, for cultivar improvement.     

Genetic diversity in cultivars and landraces of Oryza sativa subsp. indica as revealed by AFLP markers.

Genetic diversity among 49 Indian accessions of rice (Oryza sativa subsp. indica), including 29 landraces from Jeypore, 12 modern cultivars, and 8 traditional cultivars from Tamil Nadu, was investigated using AFLP markers. In total, nine primer combinations revealed 664 AFLPs, 408 of which were found to be polymorphic. The percentage of polymorphic AFLPs was approximately the same within the cultivars and landraces. Similar results were obtained when genetic diversity values were estimated using the Shannon-Weiner index of diversity. Genetic diversity was slightly higher in the modern cultivars than in the traditional cultivars from Tamil Nadu. Among the landraces from Jeypore, the lowland landraces showed the highest diversity. The present study showed that the process of breeding modern cultivars did not appear to cause significant genetic erosion in rice. Cluster analysis and the first component of principle component analysis (PCA) both showed a clear demarcation between the cultivars and landraces as separate groups, although the genetic distance between them was narrow. The modern cultivars were positioned between the landraces from Jeypore and the traditional cultivars from Tamil Nadu. The second component of PCA further separated medium and upland landraces from lowland landraces, with the lowland landraces found closest to the traditional and modern cultivars.     

The alleviation of zinc toxicity by silicon is related to zinc transport and antioxidative reactions in rice

The objective of this study is to elucidate the roles of silicon (Si) in enhancing tolerance to excess zinc (Zn) in two contrasting rice (Oryza sativa L.) cultivars: i.e. cv. TY-167 (Zn-resistant) and cv. FYY-326 (Zn-sensitive). Root morphology, antioxidant defense reactions and lipid peroxidation, and histochemical staining were examined in rice plants grown in the nutrient solutions with normal (0.15 μM) and high (2 mM) Zn supply, without or with 1.5 mM Si. Significant inhibitory effects of high Zn treatment on plant growth were observed. Total root length (TRL), total root surface area (TRSA) and total root tip amount (TRTA) of both cultivars were decreased significantly in plants treated with high Zn, whereas these root parameters were significantly increased when Zn-stressed plants were supplied with 1.5 mM Si. Supply of Si also significantly decreased Zn concentration in shoots of both cultivars, indicating lower root-to-shoot translocation of Zn. Moreover, superoxide dismutase (SOD), catalase (CAT), and asorbate peroxidase (APX) activities were increased, whereas malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) concentrations were decreased in Si-supplied plants of both Zn-sensitive and Zn-resistant rice cultivars exposed to Zn stress. These alleviative effects of Si, further confirmed by the histochemical staining methods, were more prominent in the Zn-resistant cultivar than in the Zn-sensitive one. Taken together, all these results suggest that Si-mediated alleviation of Zn toxicity is mainly attributed to Si-mediated antioxidant defense capacity and membrane integrity. The possible role of Si in reduction of root-to-shoot translocation of Zn can also be considered.     

Exposure to aflatoxin B<Subscript>1</Subscript> in Thailand by consumption of brown and color rice

This study assessed the aflatoxin B₁ (AFB₁) intake of the Thai population through consumption of contaminated brown and color rice. A total of 240 rice samples from two harvesting periods were collected in June/July 2012 (period I) and in December 2012/January 2013 (period II) and analyzed for AFB₁ by HPLC with fluorescence detection (limit of detection (LOD)?=?0.093 ng/g). Exposure assessment was based on AFB₁ levels in rice and food intake data for rice according to Thai National Consumption. Frequency and levels of AFB₁ were higher in period I (59 %, <LOD?=?26.61 μg kg^?1) than in period II (10 %, <LOD?=?3.51 μg kg^?1). Only one sample exceeded the Thai standard limit for total aflatoxin of 20 μg kg^?1, but 12 out of 240 rice samples exceeded the European Union maximum level for AFB₁ of 2 μg kg^?1. The data showed that the quality and safety of Thai rice largely comply with the requirement for both exports and domestic consumption. According to the Thai National Consumption data, the estimated AFB₁ intake via rice consumption in period I and period II was 0.80 and 0.12 μg kg^?1 bw day^?1, respectively. The potential risk for cancer, based on the recommendation of the JECFA, was estimated to be 0.011 person/year/100,000 people at a mean consumption. Although the risk via consumption of Thai rice seems to be low, the maximum levels of AFB₁ in this staple food suggest that careful monitoring and surveillance of AFB₁ contamination in rice is essential to ensure the safety of rice.     

水稻一多拷贝微卫星DNA多态性分析

用一个多拷贝微卫星 DNA标记分析了238份栽培水稻的遗传多样性及遗传多样性从农家品种到现代栽培品种的动态变化。共检测出16种长度变异类型和32种表现型,但长度变异类型数、表现型数和表现型多样性水平在农家品种和现代栽培品种间没有显著差别。该标记在遗传资源评价和DNA指纹图谱建设中有重要的应用价值,也是研究环境或人工对多拷贝基因选择作用的一个良好模型。 Abstract:The genetic diversity of 238 cultivated rice and its difference between landraces and modern cultivars have been assayed using a multiple copy microsatellite DNA marker.Although 16 length variants and 32 phenotypes were found in all accessions,no significant differences in number of length variants,number of phenotypes and phenotypic diversity were detected between landraces and modern cultivars.This marker is of great value in evaluating genetic germplasm and constructing DNA fingerprints of rice and provides an elite model for tracing effectiveness of natural and artificial selection on multiple copy genes.     

Insight into the Anti-Inflammatory Mechanism of Action of Atrial Natriuretic Peptide,a Heart Derived Peptide Hormone: Involvement of COX-2, MMPs,and NF-kB Pathways

We sought to determine the in vivo anti-inflammatory activity of atrial natriuretic peptide (ANP) using 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced acute and chronic skin inflammatory mice model. ANP treatment (2 μg/kg body weight/day/i.p. for acute and 0.5 μg/kg body weight/day/i.p. for chronic inflammatory study) was started after 30 min of TPA application. The standard drug, aspirin (ASP) (20 μg/kg body weight/day/i.p.; 10 μg/kg body weight/day/i.p., respectively) was used as a positive control for the both acute and chronic study. TPA alone treated mice exhibited a marked increase in the ear length (7 ± 0.08 vs. 13 ± 0.7 mm, p < 0.001) as well as in ear weight (80 ± 1.3 vs. 130 ± 1.5 mg, p < 0.001) as compared with control mice. Upon treatment with ANP, the increased ear length and weight were reverted back to near normal level. Similarly, ANP treatment markedly suppressed the TPA-induced chronic skin inflammatory lesion (5.7 ± 0.2 vs. 0.95 ± 0.05, p < 0.001) as compared with TPA-induced mouse skin. TPA-induced alterations in the levels of serum C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), total white blood cell (TWBC), serum tumor necrosis factor-α (TNF-α), natriuretic peptide receptor-A (NPR-A), cyclooxygenase-2 (COX-2), matrix metalloproteinase-2/-9 (MMP-2/-9) and nuclear factor kappa B (NF-κB) (p < 0.01, respectively) were reverted back to near normal levels. The results of the present study clearly show the in vivo anti-inflammatory activity of ANP, which is comparable with that of a standard drug, ASP. Our results suggest that ANP elicits its anti-inflammatory activity by down-regulating the expressions of NPR-A, COX-2, MMPs and NF-κB.     

Abundance,distribution, and secondary production of the apple snail Pomacea flagellata (Say, 1829) in Bacalar Lake,a tropical karstic system in southern Mexico

Pomacea flagellata is a gastropod conspicuous in freshwater environments, and represents a fishing resource. To assess their abundance, distribution, and secondary production, monthly samplings were carried out in Bacalar Lake from June 2012 to May 2013 at 12 sampling sites. In each site, three random transects were marked parallel to the shore. All snails on transect were collected and shell length and wet weight measured. The highest density occurred in September (1.27 ind.m^?2), lowest in October (0.47 ind.m^?2). Shell lengths ranged from 2 to 56 mm, with recruitment in January–March. Growth parameters were L_∞ 59.50 mm, K 0.65.year^?1; the lifetime span was 3 years. Average biomass reached 5.57 wet g.m^?2 and secondary production was 6.025 wet g.m^?2.year^-1; annual renewal rate P/B 1.08. Highest abundance and secondary production was contributed by individuals between 31 and 41 mm in length. A potential biomass of 25.06 tons of snails was estimated in the lake. Snail densities, secondary production, and turnover were very low during the year, indicating that it is not viable to consider a commercial catch without affecting the population. A ban of 10 years is proposed, and aquaculture practices of snails are recommended to recover the resource.     

Veronaea botryosa: Molecular Identification with Amplified Fragment Length Polymorphism (AFLP) and In vitro Antifungal Susceptibility

Inter- and intraspecific genomic variability of 18 isolates of Veronaea botryosa originating from clinical and environmental sources was studied using amplified fragment length polymorphism (AFLP). The species was originally described from the environment, but several severe cases of disseminated infection in apparently healthy individuals have been reported worldwide. All tested strains of V. botryosa, identified on the basis of sequencing and phenotypic and physiological criteria prior to our study, were confirmed by AFLP analysis, yielding a clear separation of V. botryosa as a rather homogeneous group from related species. In vitro antifungal susceptibility testing resulted in MIC₉₀s across all strains in increasing order posaconazole (0.25 μg/ml), itraconazole (1 μg/ml), voriconazole (4 μg/ml), terbinafine (4 μg/ml), caspofungin (8 μg/ml), anidulafungin (8 μg/ml), isavuconazole (16 μg/ml), amphotericin B (16 μg/ml), and fluconazole (32 μg/ml). Overall, the isolates showed a uniform pattern of low MICs of itraconazole and posaconazole, but high MICs for remaining agents. The echinocandins (caspofungin and anidulafungin) had no activity against V. botryosa. There was no statistically significant difference between susceptibilities of environmental (n = 11) and clinical (n = 7) isolates of V. botryosa (P > 0.05).     

Association mapping of agro-morphological characters among the global collection of finger millet genotypes using genomic SSR markers

Identification of alleles responsible for various agro-morphological characters is a major concern to further improve the finger millet germplasm. Forty-six genomic SSRs were used for genetic analysis and population structure analysis of a global collection of 190 finger millet genotypes and fifteen agro-morphological characters were evaluated. The overall results showed that Asian genotypes were smaller in height, smaller flag leaf length, less basal tiller number, early flowering and early maturity nature, small ear head length, and smaller in length of longest finger. The 46 SSRs yielded 90 scorable alleles and the polymorphism information content values varied from 0.292 to 0.703 at an average of 0.442. The gene diversity was in the range of 0.355 to 0.750 with an average value of 0.528. The 46 genomic SSR loci grouped the 190 finger millet genotypes into two major clusters based on their geographical origin by the both phylogenetic clustering and population structure analysis by STRUCTURE software. Association mapping of QTLs for 15 agro-morphological characters with 46 genomic SSRs resulted in identification of five markers were linked to QTLs of four traits at a significant threshold (P) level of ≤0.01 and ≤0.001. The QTL for basal tiller number was strongly associated with the locus UGEP81 at a P value of 0.001 by explaining the phenotypic variance (R ²) of 10.8 %. The QTL for days to 50 % flowering was linked by two SSR loci UGEP77 and UGEP90, explained 10 and 8.7 % of R ² respectively at a P value of 0.01. The SSR marker, FM9 found to have strong association to two agro-morphological traits, flag leaf width (P—0.001, R ²—14.1 %) and plant height (P—0.001, R ²—11.2 %). The markers linked to the QTLs for above agro-morphological characters found in the present study can be further used for cloning of the full length gene, fine mapping and their further use in the marker assisted breeding programmes for introgression of alleles into locally well adapted germplasm.     

In vitro antifungal activity of antifungalmycin 702, a new polyene macrolide antibiotic, against the rice blast fungus Magnaporthe grisea

Antifungalmycin 702, a novel polyene macrolide antibiotic produced by Streptomyces padanus JAU4234, strongly inhibited mycelial growth of the rice blast fungus, Magnaporthe grisea, with EC₅₀ of 37 μg/ml and EC₉₀ of 136 μg/ml. Significant reduction in the number of conidia was observed at above 20 μg/ml. Conidia germination and appressorium formation were also suppressed and were not viable with >40 μg/ml. When treated with antifungalmycin 702, hyphae morphology became irregular. Based on microscopic examination, antifungalmycin 702 may exert its antifungal activity by changing the structure of cell membranes and the cytoskeleton and interacting with the organelles. Antifungalmycin 702 thus has potential as a new fungicide in the treatment of rice blast disease.     

Establishment of dedifferentiated callus of haploid origin from unfertilized ovaries of tea (Camellia sinensis (L.) O. Kuntze) as a potential source of total phenolics and antioxidant activity

This is the first report of induction of haploid callus with significant antioxidant activity from unpollinated ovary cultures of tea. Out of the five cultivars tested, TV18 gave the highest percentage of callus induction. Within 1 wk of induction, ovules swelled to almost double their original size, and white, friable callus emerged. A high cytokinin/auxin ratio, provided by 8.5 μM benzyl adenine and 4.5 μM 2,4-dichlorophenxyacetic acid, and high-temperature treatment (33°C) for 10 d in the dark promoted maximum callus induction. Callus was maintained on MS medium containing 22.2 μM benzyl adenine and 9.8 μM indolebutyric acid (callus line RM 1) in the light at 25°C. Well-developed tracheids were formed within 4 wk in callus subcultured on MS medium containing 1.8 μM thidiazuron and 5.0 μM 2,3,5-triiodobenzoic acid (line RM 2). Flow cytometric analysis revealed that most cells were haploid. Both RM 1 and RM 2 produced phenolic compounds with significant antioxidant capacity. Phenolic content showed a positive linear correlation with antioxidant activity. The total phenolic content of RM 1 was 3.47?±?0.21 gallic acid equivalents (GAE) mg/g dry weight and that of RM 2 was 2.39?±?0.12 GAE mg/g dry weight. Antioxidant activity was measured using IC₅₀, a measure of inhibitory concentration; a lower IC₅₀ value reflects greater antioxidant activity. The IC₅₀ value of RM 1 was 2,530 μg/ml and that of RM 2 was 3,170 μg/ml. The results suggested that the phenolic compounds contributed significantly to the antioxidant capacity of the in vitro cell lines.     

Intracellular production of IFN-alpha 2b in Lactococcus lactis

Human interferon alpha (IFN-α) was expressed in two strains of Lactococcus lactis by aid of two promoters (P32 and Pnis) giving rise to two recombinant strains: MG:IFN and NZ:IFN, respectively. The expression of IFN was confirmed by ELISA and western blotting. Highest production was achieved using glucose for growth of both recombinant strains with nisin, used for induction of the recombinant strain with Pnis promoter, at 30 ng/ml. The optimum time for MG:IFN was 9 h and for NZ:IFN was 4.5 h. The highest productions by MG:IFN and NZ:IFN were 1.9 and 2.4 μg IFN/l, respectively. Both of the expressed IFNs showed bioactivities of 1.9 × 10⁶ IU/mg that were acceptable for further clinical studies.     

Selenium (Se) Seed Priming Induced Growth and Biochemical Changes in Wheat Under Water Deficit Conditions

Insufficient stand establishment at early growth stages in wheat (Triticum aestivum L.) due to drought stress is a major problem that limits overall efficiency and yield of crop. Priming of seed is an effective method for raising seed performance and improving tolerance of crops to abiotic stresses especially drought. The seeds of two local wheat cultivars (Kohistan-97 and Pasban-90) were soaked in distilled water or sodium selenate solutions of 25, 50, 75, and 100 μM for 1/2 or 1 h at 25 °C and later re-dried to their original moisture levels before sowing. One-hour priming significantly increased root length stress tolerance index, dry matter stress tolerance index, and total biomass of seedlings; however, no significant effect of changing duration of Se seed priming was observed on plant height stress tolerance index and shoot/root ratio. Among cultivars, Kohistan-97 was found to be more responsive to Se seed treatment as 1 h priming at 100 μM significantly increased its total biomass by 43 % as compared to control treatment. Although biomass of seedlings was not affected with Se seed priming under normal conditions, but it increased significantly with increase in rates of Se under drought stress conditions. One-hour priming at 75 μM increased the total sugar content and total free amino acids in both wheat cultivars. A more significant decrease in soluble proteins of seedlings was observed by 1 h priming than 1/2 h priming under drought stress conditions.     

Pre-harvest aflatoxins and <Emphasis Type="Italic">Aspergillus flavus</Emphasis> contamination in variable germplasms of red chillies from Kunri,Pakistan

Various cultivars of red chilli were collected from a small town named Kunri, located in the province Sindh, Pakistan. This town is a hub of red chilli production in Asia. A total of 69 samples belonging to 6 cultivars were obtained and analysed for the occurrence of aflatoxins and Aspergillus flavus, to explore the potential of resistant and susceptible germplasm. Aflatoxins were detected by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), while A. flavus was isolated and identified using agar plate, blotter paper, deep freezing and dilution techniques. Molecular characterization using internal transcribed spacer (ITS) 1/4 and A. flavus specific FL1-F/R primers confirmed the identity of A. flavus. The data revealed that 67 and 75% samples contaminated with aflatoxin B₁ (AFB₁) and with A. flavus, respectively. A highly susceptible chilli cultivar was ‘Nagina’, showing 78.8% frequency of total aflatoxins (1.2–600 μg/kg) and a mean of 87.7 μg/kg for AFB₁ and 121.9 μg/kg for total aflatoxins. A. flavus was detected with 93% frequency and 2.14 × 10⁴ colony forming units. In contrast, cultivars ‘Kunri’ and ‘Drooping Type’ were found to be resistant, with low levels of aflatoxins and fungal counts. The study was conducted for the first time to explore two potential cultivars that were less susceptible towards A. flavus and aflatoxin contamination. These cultivars could be preferably cultivated and thereby boost Pakistan’s chilli production.