Efficient utilization of date palm waste for the bioethanol production through Saccharomyces cerevisiae strain.

Dates (Phoenix dactylifera L.) are rich in nutritional compounds, particularly in sugars. Sugars offer anaerobic fermentation, used for bioethanol production. Recently, researchers and industrialists finding ways to produce low-cost bioethanol on large scale using agricultural wastes. Date palm residual is the largest agricultural waste in Pakistan, which can be the cheapest source for bioethanol production, whereas the current study was designed to explore the possible utilization and the potential of date palm waste for bioethanol production through Saccharomyces cerevisiae grown in yeast extract, Bacto peptone, and d-glucose medium. The fermentation process resulted in the production of 15% (v/v) ethanol under the optimum condition of an incubation period of 72 hr and three sugars (glucose, fructose, and sucrose) were found in date waste. The functional group of ethanol (C2H5OH) was also found via Fourier-transform infrared spectroscopy (FTIR) analysis. Therefore, S. cerevisiae could be recommended for ethanol production due to short fermentation time at 25% inoculum in 30°C and reduced the processing cost. Common date varieties of low market value are a preferred substrate for the process of producing industrial ethanol. Additionally, proximate analysis of date fruit by near-infrared spectroscopy revealed moisture contents (16.84%), crude protein (0.3%), ash (9.8%), crude fat (2.6%), and neutral detergent fibers (13.4%). So, date fruit contains various nutrients for microbial growth for ethanol production.

COTIP: Cotton TILLING Platform, a Resource for Plant Improvement and Reverse Genetic Studies.

Cotton is cultivated worldwide for its white fiber, of which around 90% is tetraploid upland cotton (Gossypium hirsutum L.) carrying both A and D genome. Since centuries, yield increasing efforts for the cotton crop by conventional breeding approaches have caused an extensive erosion of natural genetic variability. Mutation based improvement strategies provide an effective way of creating new allelic variations. Targeting Induced Local Lesions IN Genomes (TILLING) provides a mutation based reverse genetic strategy to create and evaluate induced genetic variability at DNA level. Here, we report development and testing of TILLING populations of allotetraploid cotton (G. hirsutum) for functional genomic studies and mutation based enrichment of cotton genetic resources. Seed of two cotton cultivars "PB-899 and PB-900" were mutagenized with 0.3 and 0.2% (v/v) ethyl methanesulfonate, respectively. The phenotyping of M1 and M2 populations presented numerous mutants regarding the branching pattern, leaf morphology, disease resistance, photosynthetic lesions and flower sterility. Molecular screening for point mutations was performed by TILLING PCR aided CEL1 mismatch cleavage. To estimate the mutation frequency in the mutant genomes, five gene classes were TILLed in 8000 M2 plants of each var. "PB-899" and "PB-900." These include actin (GhACT), Pectin Methyl Esterase (GhPME), sucrose synthase (GhSUS), resistance gene analog, and defense response gene (DRGs). The var. PB-899 was harboring 47% higher mutation induction rate than PB-900. The highest rate of mutation frequency was identified for NAC-TF5 (EU706348) of DRGs class, ranging from 1/58 kb in PB-899 to 1/105 kb in PB-900. The mutation screening assay revealed the presence of significant proportion of induced mutations in cotton TILLING populations such as 1/153 kb and 1/326 kb in var. "PB-899" and "PB-900," respectively. The establishment of a cotton TILLING platform (COTIP) and data obtained from the resource TILLING population suggest its effectiveness in widening the genetic bases of cotton for improvement and utilizing it for subsequent reverse genetic studies of various genes.

Development of molecular method for sex identification in date palm (Phoenix dactylifera L.) plantlets using novel sex-linked microsatellite markers.

Microsatellite markers containing simple sequence repeats (SSRs) are a valuable tool for genetic analysis. Date palm is a dioecious and slow flowering and is very difficult to identify the gender of the trees until it reaches the reproductive age (5-10 years). A total of 12 microsatellite primers were used with 30 date palm samples, 14 parents (8 male + 6 females) and 16 progeny (developed from parents breeding) which showed that microsatellites were highly polymorphic, having a great number of alleles. A total of 124 alleles were characterized in 12 SSR loci. On average, there are 9.08 alleles per locus, with a range from 5 to 16 alleles, for primers mpdCIR15 and mpdCIR57, respectively. These primers produced 15 polymorphic loci specifically in male date palm samples and the seedlings harboring the unique fragments were further characterized as male plants. Increasingly, 38.46 % of these loci were scored as homozygous alleles while 61.53 % heterozygous allelic loci were determined. Primer mpdCIR48 produced a specific locus (250/250) in all male samples whereas the same locus was absent in female samples. Similarly, a locus of 300/310 bp reoccurred in 5 date palm male samples using marker DP-168 which indicated that these are the promising candidate marker to detect the sex in date palm seedlings at early stage. The data resulted from combination of 12 primers enabled the 16 seedling samples progeny (developed from parents breeding) of date palm cultivars to divide into two groups i.e., male and female regarding their sex expression comparative to the parents (male + female) using the principle coordinate analysis.

Detection of citrus huanglongbing-associated 'Candidatus Liberibacter asiaticus' in citrus and Diaphorina citri in Pakistan, seasonal variability, and implications for disease management.

We report the detection of the huanglongbing (HLB)-associated bacterium 'Candidatus Liberibacter asiaticus' from both plants and insects in Pakistan and the seasonal variability in the numbers of 'Ca. L. asiaticus'-positive psyllid vector, Diaphorina citri. Our studies showed that 'Ca. L. asiaticus' was detectable from trees in areas with maximum temperatures reaching nearly 50°C (average maximum of 42°C). However, the bacterium was present at very low levels in psyllids both in summer (June to August) and autumn (September to November) in contrast to reports from Florida, where the bacterium was detectable at very high levels during October to November. We hypothesize that hot summer temperatures in Pakistan may interfere with acquisition and replication of 'Ca. L. asiaticus' in psyllids and may lead to dead or non transmissible 'Ca. L. asiaticus' in plants. Psyllid counts were very low in both summer and winter, showed a population peak ('Ca. L. asiaticus'-positive vectors) in spring, and showed a larger peak ('Ca. L. asiaticus'-free psyllids) in autumn. Natural thermotherapy during hot summers and a low vector population during environmental extremes may have played a major role in long-term survival of the citrus industry in Pakistan. These results may be useful in developing management strategies for U.S. citrus industries in Texas and California.

A comparative study of interaction of ibuprofen with biocompatible polymers.

In this paper we are reporting the interaction of a non-steroidal anti-inflammatory drug ibuprofen (IBF) with various biocompatible polymers. Being amphiphilic, the drug interacts with the polymers similar to the interaction of surfactants and polymers. Therefore, we have considered the polymer-amphiphile interaction approach using conductimetry. The polymers of different charges (cationic, anionic, and nonionic) have been taken for the study. It was found that the critical aggregation concentration (cac) decreases on increasing the polymer concentrations of cationic as well as nonionic polymers whereas it increases for anionic polymers. The results imply that anionic IBF interacts with cationic and nonionic polymers more strongly as compared to the anionic polymers. A possible anionic-anionic repulsion is responsible for the weak interaction of IBF with anionic polymers. On the other side, the critical micelle concentration (cmc) increases for all polymers which is a usual indication of the interaction between amphiphiles and polymers. Free energies of aggregation (ΔG(agg)) and micellization (ΔG(mic)) were also computed with the help of degrees of micelle ionization obtained from the specific conductivity - [IBF] isotherms.