Characterization of Drought Tolerance in Bread Wheat Genotypes Using Physiological Indices

Wheat (Triticum aestivum L.) is a staple food in many countries and is regarded as a vital source of nutrition. Drought is one the most prevalent limitations to wheat growth and development. Herein a two year study was conducted using 25 diverse wheat genotypes obtained from the gene pool of various research institutes of Pakistan to characterize their drought tolerance using various physiological indices like relative water content (RWC), relative dry weight (RDW), water saturation deficit (WSD), relative water loss (RWL), flag leaf area (LA), chlorophyll content index (CC) and their association with the grain yield (GY). Analysis of variance (ANOVA) indicated the presence of significant amount of differences and genetic diversity among genotypes under study. Correlation analysis exposed positive association of CC and LA with GY. However, RWC was shown to have a highly significant and negative association with WSD and RWL. Principal component analysis (PCA) showed that out of the 7 PCs only 2 were significant having eigenvalues >?1; cumulatively accounting for 88.70% and 73.03% of the total variation under control and drought stress conditions, respectively. Strikingly the results of the PCA biplots and cluster heat map exposed G1 (Barani-17), G2 (Dharabi-11), G3 (Ehsan-16), G4 (Chakwal-50), G17 (Ujala-2016) and G23 (Kohistan-97) as potential drought tolerant genotypes. Selection of the positively associated indices would be fruitful and the tolerant genotypes having drought tolerance potential could be utilized in future wheat breeding programs to develop high yielding and drought tolerant genotypes.

     

Boron-Calcium Synergically Alleviates Aluminum Toxicity in Wheat Plants (Triticum aestivum L.)

The effects of B and Ca treatments on root growth, nutrient localization and cell wall properties in wheat ( Triticum aestivum L.) plants with and without Al stress were investigated. Seedlings were grown hydroponically in a complete nutrient solution for 7 d and then treated with B (0, 40 μM), Ca (0, 2,500 μM), and Al (0, 100 μM) in a 500 μM CaCl₂ solution for 8 d. The cell wall materials (CWM) were extracted with a phenol: acetic acid: water (2:1:1 w/v/v) solution and used for subsequent pectin extraction with trans -1,2-diami-nocyclohexane- N,N,N,N -tetraacetic acid (CDTA) and Na₂CO₃ solutions. Boron, Ca, and B + Ca treatments enhanced root growth by 19.5, 15.2, and 27.2%, respectively, compared to the control (pH 4.5). Calcium and B+Ca treatments enhanced root growth with Al stress by 43 and 54%, respectively, while B did not exert any effect. The amounts of CWM and pectin per unit of root fresh weight increased by Al treatment, whereas the Ca and B+Ca treatments slightly reduced the contents of these components. Seventy-four percent of total B, 69% of total Ca, and 85% of total Al were located in the cell wall in the B, Ca, and Al treatments, respectively and 32% of total B, 33% of total Ca, and 33% of total Al were located in the CDTA-soluble and Na₂CO₃-soluble pectin fractions. A more conspicuous localization of B was observed in the presence of Al. Aluminum treatment markedly decreased the Ca content in the cell wall as well as pectin fractions, mainly in the case of the CDTA-soluble pectin fraction. Boron + Ca treatment decreased the Al content in the cell wall and pectin fractions compared to the Ca treatment alone in the presence of Al. It is concluded that the B+Ca treatment enhanced root growth and, B and Ca uptake, and helped to maintain a normal B and Ca metabolism in the cell walls even in the presence of Al.     

Effect of dietary oxidized oil and vitamin E on growth performance,lipid peroxidation and fatty acid profile of Labeo rohita fingerlings

A 3 × 3 factorial experiment was conducted on Labeo rohita fingerlings to evaluate the effect of dietary oxidized oil and vitamin E. Nine experimental diets were made, based on three degrees of oil oxidation (fresh oil, low oil oxidation and high oil oxidation), and each level of oxidation was further supplemented with three levels of vitamin E (0, 100 and 1,000 mg/kg). Weight gain% and specific growth rate (SGR) of fish fed fresh fish oil and low oil oxidation level were significantly higher than highly oxidized oil. Moreover, vitamin E supplemented fish also showed better growth performance. Oil oxidation caused a significant reduction in the concentrations of α‐tocopherol and increase in TBARS level and antioxidant enzyme activities in fish liver and muscles. However, increasing the dietary vitamin E abrogated these effects. Dietary vitamin E supplementation improved the fatty acid, more specifically polyunsaturated fatty acids profile of oxidized oil fed fish. In conclusion, dietary oxidized fish oil increased the oxidative stress condition of fish but supplementation of high dose of vitamin E prevented lipid oxidation, improved growth performance and fatty acid profile of L. rohita.     

Assessment of gut microbiota in different developmental stages of Malaysian Mahseer (Tor tambroides)

Malaysian Mahseer (Tor tambroides) has a good prospect for aquaculture because of its high market demand. However, there is a scarce information on gut microbiota associated with Malaysian Mahseer unlike other fish species. Therefore, we constructed and compared gut microbiota in different developmental stages (larval, juvenile, fingerling, yearling, and adult) using culture dependent and PCR‐DGGE fingerprinting technique for better understanding of gut microbiota composition associated with T. tambroides. Culturable gut microbiota composition in all developmental stages were composed of β‐ and γ‐Proteobacteria, and Bacilli. Biodiversity analysis of culturable gut microbiota showed that larval, juvenile, and adult stages have higher diversity than fingerling and yearling stages. Ward's linkage cluster analysis showed that culturable gut microbiota composition in larval and juvenile stages were close to adult stages, whereas fingerling and yearling stage composed same cluster. PCR‐DGGE fingerprinting technique showed that unculturable gut microbiota were constituted by α‐and γ‐Proteobacteria, and Actinobacteria. Ward's linkage cluster analysis showed that unculturable gut microbiota composition in both larval and juvenile stages were distinct from other developmental stages. Our results revealed that gut microbiota composition were varied in different developmental stages of Malaysian Mahseer and continuous shifts of gut microbiota from larval to adult stages.     

Production of polyclonal antiserum against recombinant VP28 protein and its application for the detection of white spot syndrome virus in crustaceans

The VP28 gene of white spot syndrome virus (WSSV) was cloned into pRSET B expression vector. The VP28 protein was expressed as a protein with a 6-histidine taq in Escherichia coli GJ1158 with NaCl induction. Antiserum was raised against this recombinant-VP28 protein in rabbits and it recognized VP28 protein in naturally and experimentally WSSV-infected shrimp, marine crabs, freshwater prawns and freshwater crabs. The antiserum did not recognize any of the other known WSSV structural proteins. Various organs such as eyestalks, head muscle, gill tissue, heart tissue, haemolymph, tail tissue and appendages were found to be good materials for detection of WSSV using the antiserum and detection of WSSV was successful in experimentally infected Penaeus monodon and P. indicus at 12 and 24 h post-infection (p.i.), respectively. The antiserum was capable of detecting WSSV in 5 ng of total haemolymph protein from WSSV-infected shrimp.     

Improved agar plate assays of bovine lysozyme and haemolytic complement activity

Bovine serum and colostral whey samples were examined for lysozyme and haemolytic complement activity, employing agar plate techniques. The tests were carried out in agarose gel containing Micrococcus lysodeikticus (for lysozyme), and antibody sensitized rabbit erythrocytes (for complement), respectively. The confirmation of lysozyme (E.C.3.2.1.17) — dependent lysis has been presented elsewhere (Lie & Syed 1986), while heat-inaotivation and antibody to C3 were used in the present study to confirm that the haemolytic activity was attributable to the complement cascade. Repeatability and sensitivity of the described tests were found to be superior to those of photometric procedures. Staining and preservation techniques were developed which extended the applicability of the assay, as they made reading of results independent of time and resulted in the plates being very suitable for photography and storage.     

Simple protoplast isolation system for gene expression and protein interaction studies in pineapple (Ananas comosus L.)

Background

Seed viability monitoring is very important in ex situ germplasm preservation to detect germplasm deterioration. This requires seed-, time- and labor- saving methods with high precision to assess seed germination as viability. Although the current non-invasive, rapid, sensing methods (NRSs) are time- and labor-saving, they lack the precision and simplicity which are the virtues of traditional germination. Moreover, they consume a considerable amount of seeds to adjust sensed signals to germination percentage, which disregards the seed-saving objective. This becomes particularly severe for rare or endangered species whose seeds are already scarce. Here we propose a new method that is precise, low-invasive, simple, and quick, which involves analyzing the pattern of dehiscence (seed coat rupture), followed by embryonic protrusion.

Results

Dehiscence proved simple to identify. After the trial of 20 treatments from 3 rice varieties, we recognized that dehiscence percentage at the 48th hour of germination (D(48)) correlates significantly with germination rate for tested seed lots. In addition, we found that the final germination percentage corresponded to D(48) plus 5. More than 70% of the seeds survived post-dehiscence desiccation for storage. Hydrogen peroxide (1 mM) as the solution for imbibition could further improve the survival. The method also worked quicker than tetrazolium which is honored as a fast, traditional method, in detecting less vigorous but viable seeds.

Conclusion

We demonstrated the comprehensive virtues of dehiscence method in assessing rice seed: it is more precise and easier to use than NRSs and is faster and more seed-saving than traditional methods. We anticipate modifications including artificial intelligence to extend our method to increasingly diverse circumstances and species.

     

Identification of Aspergillus flavus and aflatoxin in home mix layer poultry feed in relation to seasons in Karachi,Pakistan

Fungal toxins in feed are leading issue in poultry industry causing a detrimental effect on the performance and health of poultry. The study was carried out to determine the incidence and concentration of the aflatoxins and their major producer Aspergillus flavus in home mix layer poultry feed in respect of seasonal variation throughout the year. A total of (n?= 204) home mix poultry layer feed samples were analyzed for the isolation of fungi. The isolates were initially screened through colony morphology and microscopic examination. However, aflatoxin concentration was determined by ELISA. Revealed results indicated that, the highest percentage of A. flavus was found during the months of June to August 50/54 (92.5%) followed by September to November 43/65 (66.1%), March to May 21/40 (52.5%), and December to February 18/45 (40%). As a whole, the incidence was recorded 132/204 (64.7%). Moreover, of the 132 samples, 41 (31%) were exceeded in respect of aflatoxin contamination from the legal limit (20 μg/kg) imposed by Food Drug Association (FDA). Statistically, the growth of A. flavus and aflatoxin production was found significantly different in respect of seasonal variation. As highest total viable fungal count (9.9?×?10⁴ CFU/g) and aflatoxin level (72.27 μg/kg) were recorded during the months of June to August and lowest in December to February. Consequently, instantaneous essential control measures are demanded regarding appropriate storage and adequate drying in post-harvesting season. Along with surveillance plans and austere regulations for monitoring the aflatoxin contents for the wellbeing of consumers.