Enhancement of germination and yield of cotton through optical seed priming: Lab. and diverse environment studies.

The current study demonstrates the practical application of optical seed priming technology to improve cotton seed germination, plant growth, crop yield, and fiber quality. The hypothesis of this study is that seed irradiation with different colors of light can improve germination and cotton productivity in different environments. In the priming of cotton seeds, a wider range of the light spectrum was used, ranging from ultraviolet (UV) to red wavelengths. Various light sources such as blue LED, red LED, diode laser, UV-B, and UV-C were studied, along with different exposure times and energy densities. The exposure time ranged from 1.0 to 36.0 minutes, while the energy density doses varied from 88 to 7550 mJ cm-2, depending on the light source. In laboratory conditions, the investigation on the impact of optical seed priming on germination showed a maximum improvement of up to 180% compared to the control group. Among the different light sources and energy densities, blue LED light was found to be the most effective for enhancing cotton seed germination across different varieties. To validate the findings from the laboratory, large-scale field trials were conducted in two different environments in Pakistan, namely Tandojam and Faisalabad. The field trials demonstrated significant improvements in germination and yield, with increases of up to 37% and 74% over the control group, respectively. Once again, blue LED light emerged as the best light source for optical seed priming at the farm level. These field trials provided encouraging results, indicating the potential of the eco-friendly optical seed priming technique. The study suggests that optical seed priming can be a commercially viable technology for improving cotton seed germination, plant growth, crop yield, and fiber quality. By utilizing this technique, growers and researchers in developing countries can address the challenge of poor cotton germination and potentially enhance their agricultural productivity.

Early detection of stripe rust infection in wheat using light-induced fluorescence spectroscopy.

In the current study, the application of fluorescence spectroscopy along with the advanced statistical technique and confocal microscopy was investigated for the early detection of stripe rust infection in wheat grown under field conditions. The indigenously developed Fluorosensor fitted with LED, emitting monochromatic light was used that covered comparatively larger leaf area for recording fluorescence data thus presenting more reliable current status of the leaf. The examined leaf samples covered the entire range of stripe rust disease infection from no visible symptoms to the complete disease prevalence. The molecular changes were also assessed in the leaves as the disease progresses. The emission spectra mainly produce two fluorescence emission classes, namely the blue-green fluorescence (400-600 nm range) and chlorophyll fluorescence (650-800 nm range). The chlorophyll fluorescence region showed lower chlorophyll bands both at 685 and 735 nm in the asymptomatic (early diseased) and symptomatic (diseased) leaf samples than the healthy ones as a result of partial deactivation of PSII reaction centers. The 735 nm chlorophyll fluorescence band was either slight or completely absent in the leaf samples with lower to higher disease incidence and thus differentiate between the healthy and the infected leaf samples. The Hydroxycinnamic acids (caffeic and sinapic acids) showed decreasing trend, whereas the ferulic acid increased with the rise in disease infection. Peak broadening/shifting has been observed in case of ferulic acid and carotenes/carotenoids, with the increase in the disease intensity. While using the LEDs (365 nm), the peak broadening and the decline in the chlorophyll fluorescence bands could be used for the early prediction of stripe rust disease in wheat crop. The PLSR statistical techniques discriminated well between the healthy and the diseased samples, thus showed promise in early disease detection. Confocal microscopy confirmed the early prevalence of stripe rust disease infection in a susceptible variety at a stage when the disease is not detectable visually. It is inferred that fluorescence emission spectroscopy along with the chemometrics aided in the effective and timely diagnosis of plant diseases and the detected signatures provide the basis for remote sensing.

Phenolic Profile and Thermal Stability of Monovarietal Extra Virgin Olive Oils Based on Synchronous Fluorescence Spectroscopy.

The olive oil production in Pakistan has recently been started with the cultivation of exotic cultivars that are successfully adapted at Barani Agriculture Research center (BARI), Chakwal, Pakistan in Potohar valley. Therefore, characterization of extra virgin olive oil (EVOO) from this agro-climatic region is mandatory in establishing its biochemical profile and thermal stability. Seventeen monovarietal EVOOs extracted from these cultivars were analysed using synchronous fluorescence spectroscopy (SFS) and subjected to heating at 115, 150 and 170 °C for 15 min to identify their thermal stability. SFS emission spectra differentiated EVOOs on the basis of phenolic compounds that are denatured at high temperature, further chlorophyll contents also decreased with increasing temperature. The strong emission at ca. 351 nm, suggested to be vanillic acid, 391-471 nm for blue green region (BGR) assigned to other phenolic compounds and two peaks at 672 and 723 nm for chlorophyll became the bases for grouping through Hierarchical clustering. Most of the EVOOs were stable at 150 °C but showed denatured spectra at 170 °C, the only EVOO extracted from Spanish cultivar Arbequina was found to have moderate fluorescence emission from both vanillic acid and BGR that are more likely to impart oxidative stability even after heating at 170 °C, also confirmed by lowest values of specific extinction co-efficient (K232 and K270). Moreover, variation in phenolic contents of Arbequina EVOO was observed with different harvesting stages and the early harvested olives produced more thermally stable oil as compared to late harvested olives. Arbequina oil grown in Pakistan can be better suited for cooking at high temperatures, moreover can be blended with other monovarietal EVOOs to enhance the nutritional benefits and thermal stability.

Application of Fluorescence Spectroscopy in Wheat Crop: Early Disease Detection and Associated Molecular Changes.

The application of fluorescence spectroscopy combined with chemometrics was explored in the current study for the detection of stripe rust in wheat. The healthy and stripe rust leaves were collected from the disease screening nursery. The variations in the blue-green region and chlorophyll fluorescence intensity in leaves provides the basis for the detection of stripe rust infection. With the progress of disease, the variations in the synchronous fluorescence spectroscopy (SFS) spectrum was witnessed. SFS is an excellent tool for the simultaneous measurement of multiple compound samples, in case of plants it generates evidence regarding the occurrence of leaf fluorophore bands thus revealing the biochemical variations going on at different infection stages. Based on the results of the current study, it is inferred that p-coumaric acid has the highest intensity in healthy samples followed by the asymptomatic leaf samples, whereas the band intensity of α-tocopherol, sinapic acid, chlorogenic acid, ferulic acid, tannins, flavonoid, carotenoids and anthocyanins increases in the diseased and the asymptomatic samples accordingly to the rust infection. Principal component analysis (PCA) beautifully differentiated the healthy and the infected leaf samples. It is evident that the asymptomatic samples are grouped with the diseased samples or independently; indicating the start of disease infection, the decision that is hard to make with the visual assessments. The results of the current study suggest that the fluorescence emission and the SFS spectral signatures acquired for stripe rust could be utilized as fingerprints for early disease detection.

Laser-induced fluorescence spectroscopy for early disease detection in grapefruit plants.

Biotic and abiotic stress both cause a considerable decrease in the chlorophyll content in plant leaves, which provides a means for the early diagnosis of diseases in plants. The emergence of diseases affects the fluorescence of phenolic compounds and chlorophyll, which have emissions located at 530, 686 and 735 nm. Herein, it was found that the intensity of the emission band of phenolic compounds at 530 nm increased and that of chlorophyll at 735 nm decreased with the onset of diseases. Statistical analysis through principal component analysis (PCA) and partial least squares regression (PLSR) was performed, which differentiated between apparently healthy leaf sites and diseased leaves, providing a basis for the detection of diseases in the early stages. The PLSR model was validated through the coefficient of determination (R2), standard error of prediction (SEP) and standard error of calibration (SEC) with the values of 0.99, 0.394 and 0.0.401, respectively, which authenticated the model. The prediction accuracy of the model was evaluated through root mean square error in prediction (RMSEP), with a value of 0.14, by predicting 22 unknown emission spectra of different leaf sites. Both the PCA and PLSR models produced similar results, proving that fluorescence spectroscopy is an excellent tool for early disease detection in plants.

Selection and screening of drought tolerant high yielding chickpea genotypes based on physio-biochemical indices and multi-environmental yield trials.

BACKGROUND: Chickpea is one of the major legume crops being cultivated in the arid and semi-arid regions of Pakistan. It is mainly grown on the marginal areas where, terminal drought stress is one of the serious threats to its productivity. For defining the appropriate selection criteria for screening drought tolerant chickpea genotypes, present study was conducted. Distinct chickpea germplasm was collected from different pulses breeding institutes of Pakistan and evaluated for drought tolerance at germination and early seedling stages, furthermore, at late vegetative growth stages physiochemical traits and multi-environment yield performance were also tested. RESULTS: Chickpea genotypes under different environments, were significantly varied for different seedling traits, physio-chemical attributes and seed yield. Genotypes showing drought tolerance by performing better at an early seedling stages were not correspondingly high yielding. Screening for drought tolerance on seed yield basis is the most appropriate trait to develop the drought tolerant as well as high yielding chickpea genotypes. Results confirmed that traits of early growth stages were not reflecting the drought tolerance at terminal growth stages and also did not confer high yielding. NIAB-rain fed environment proved ideal in nature to screen the chickpea genotypes whereas, NIAB-lysimeter and Kalur Kot was least effective for selecting genotypes with high seed yield. Genotypes D0091-10, K010-10, D0085-10, K005-10, D0078-10, 08AG016, 08AG004, D0080-10, 09AG002, K002-10 and D0099-10 were high yielding and drought tolerant based on their performance across multiple hotspot environments. CONCLUSIONS: The selected genotypes are intended for further evaluation for varietal approval to recommend for general cultivation on farmer fields in drought hit areas of Pakistan. Among physio-biochemical traits, higher proline, glycine betain, RWC and CMS were reflecting the higher capability to tolerate the drought stress in chickpea. Drought sensitive genotypes (K0037-10, 2204, K0052-10, 09AG015, K0042-10, CM709/06, K0068-10, K004-10, K0026-10 and K0063-10) were also identified in present study which were resourceful asset for using as contrasting parents in hybridization programs. To our knowledge, this is first report using an integrated approach involving, physio-biochemical indices, and multi-environmental yield trials, for comparison, screening and selection of chickpea genotypes for drought tolerance.

Characterization of Desi Ghee Extracted by Different Methods Using Fluorescence Spectroscopy.

In the current study, the effect of ghee extraction methods (direct cream DC, milk butter MB and milk skin MS) on its molecular composition has been investigated using Fluorescence spectroscopy. The excitation wavelength of 300 nm was found the best to produce pronounced spectral signatures of beta-carotene, vitamins and conjugated linoleic acid (CLA) in both cow and buffalo ghee types. Principal component analysis (PCA) has been applied on the spectral data to visualize the classification among ghee samples extracted by three methods. Both cow and buffalo ghee contain spectral signatures of vitamin A, E, K, D and CLA which has been verified through plotting loading vectors. The analysis of loading plots has been suggested that for cow ghee, MS extraction method conserve relatively higher concentration of beta carotene while DC and MB methods are a good choice for preserving relatively more concentrations of vitamins D, E and K. Similarly, for buffalo ghee, MS extraction method appear with higher concentration of CLA, whereas DC extraction method looks to preserve relatively higher concentration of vitamin A while MB method retains relatively low concentration of CLA and vitamins as compared to other two methods.

Thermal Effects on Biochemical Signatures of UHT, Pasteurized and Domestically Boiled Buffalo Milk Detected by Synchronous Fluorescence Spectroscopy.

Thermal treatment of milk is performed to limit bacterial growth and make it safe for human consumption. To increase the shelf life of milk, either ultrahigh temperature (UHT) or pasteurization techniques are employed that destroy the microorganisms. In this study, the synchronous front face fluorescence spectroscopy was employed for comparative study of raw, UHT treated, pasteurized and conventionally boiled milk at 93 °C (domestic boiling). Principal Component analysis clearly showed distinct clustering of UHT milk due to formation of Maillard reaction products. Fluorescence emission peak at 410 nm showed irreversible change in peak intensity attributed to conformational changes in protein due to UHT treatment while pasteurization and domestic boiling showed reversible changes when milk was cooled down to 10 °C. Furthermore, fluorescence emission peaks at 410 nm previously assigned to vitamin A has also been discussed.

Determination of curcuminoid content in turmeric using fluorescence spectroscopy.

The potential of fluorescence spectroscopy is exploited for the characterization and comparison of different turmeric varieties based on curcuminoids content in turmeric powders. Fluorescence spectra from turmeric powders has been acquired by using excitation wavelengths from 300 to 470 nm with step of 10 nm to investigate the effect of excitation wavelengths on the emission of valuable ingredients for their characterization. Emission spectra revealed that fresh wet turmeric rhizomes show emission bands at 571 nm which is due to curcumin. It is found that main ingredient of turmeric powder is curcumin and best excitation wavelength is 467 nm for its maximum emission intensity. High Pressure Liquid Chromatography (HPLC) was used as alternate standard technique for determination of curcuminoid content in the reference samples. The curcumin content in the commercially available local turmeric brands were also evaluated, one brand showed significant covariance from standard fluorescent spectra of turmeric meaning this particular brand contained minimum curcumin content or have been severely adultered. In the next step the powders were heated at different temperatures from 60 °C to 150 °C (Normal cooking & frying temperatures) to observe the difference in emission spectra particularly keeping in view the molecular composition and curcuminoid content in turmeric. The results indicate that curcumin content gradually decreases above 90 °C. Principal component analysis (PCA) has been employed on all the data to statistically differentiate small molecular changes and adulteration by covariance calculations.

Phosphate solubilizing bacteria with glucose dehydrogenase gene for phosphorus uptake and beneficial effects on wheat.

The aim of this study was to isolate, characterize and use phosphate solubilizing bacteria to enhance the bioavailability of insoluble Ca-phosphate for wheat plants. For this purpose, 15 phosphorus solubilizing bacteria (PSB) were isolated from wheat rhizospheric soils of Peshawar and southern Punjab region, Pakistan. These isolates were identified using light microscopy and 16S rRNA gene. Among the isolated bacteria, two strains (Pseudomonas sp. MS16 and Enterobacter sp. MS32) were the efficient P solubilizers based on their P solubilization activity determined qualitatively (solubilization index 3.2-5.8) as well as quantitatively (136-280 µg mL-1). These two strains produced indole-3-acetic acid (25.6-28.1 µg mL-1), gibberellic acid (2.5-11.8), solubilized zinc compounds (SI 2.8-3.3) and showed nitrogenase and 1-Aminocyclopropane-1-carboxylic acid deaminase activity in vitro. Phosphate solubilization activity of Pseudomonas sp. MS16 was further validated by amplification, sequencing and phylogenetic analysis of glucose dehydrogenase (gcd) gene (LT908484) responsible for P solubilization. Response Surface Methodology for large-scale production was used to find optimal conditions (Temperature 22.5°C, pH 7) for P solubilization. Glucose was found to support higher P solubilization in vitro. In an in vitro experiment, PSB treated wheat seedlings improved germination and seedling vigor (11% increases) as compared to un-inoculated control. Rhizoscanning of these seedlings showed an increase in various root growth parameters. Wheat inoculation with selected strain MS16 showed pronounced effect on grain yield in pot (38.5% increase) and field (17-18% increase) experiments compared to non-inoculated control. Root colonization by PSB through Florescent in situ Hybridization and Confocal Laser Scanning Microscopy confirmed their rhizosphere competence in soil. BOX-PCR confirmed the re-isolated colonies of Pseudomonas sp. MS16. The results indicated that gluconic acid producing Pseudomonas sp. MS16 from un-explored soils may be cost effective and environment friendly candidate to improve plant growth and phosphorous uptake by wheat plants.