The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) has been reported for precise genome modification in many plants. In the current study, we demonstrate a successful mutation in phytoene desaturase (RAS-PDS) of banana cv. Rasthali using the CRISPR/Cas9 system. Two PDS genes were isolated from Rasthali (RAS-PDS1 and RAS-PDS2), and their protein sequence analysis confirmed that both PDS comprises conserved motifs for enzyme activity. Phylogenetic analysis of RAS-PDS1 and RAS-PDS2 revealed a close evolutionary relationship with other monocot species. The tissue-specific expression profile of RAS-PDS1 and RAS-PDS2 in Rasthali suggested differential regulation of the genes. A single 19-bp guide RNA (gRNA) was designed to target the conserved region of these two RAS-PDS and transformed with Cas9 in embryogenic cell suspension (ECS) cultures of cv. Rasthali. Complete albino and variegated phenotype were observed among regenerated plantlets. DNA sequencing of 13 plants confirmed the indels with 59% mutation frequency in RAS-PDS, suggesting activation of the non-homologous end-joining (NHEJ) pathway. The majority of mutations were either insertion (1-5) or deletion (1-4) of nucleotides near to protospacer adjacent motif (PAM). These mutations have created stop codons in RAS-PDS sequences which suggest premature termination of RAS-PDS protein synthesis. The decreased chlorophyll and total carotenoid contents were detected in mutant lines that revealed the functional disruption of both RAS-PDS genes. Our results demonstrate that genome editing through CRISPR/Cas9 can be applied as an efficient tool for banana genome modification.

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Background:Abnormal uterine bleeding (AUB) is a common problem for which women seek gynecological consultation. Endometrial aspiration cytology (EAC) is an acceptable and valuable diagnostic procedure for screening the endometrial status.Materials and Methods:Endometrial aspiration using a menstrual regulation (MR) syringe and a 4 mm Karman's cannula was performed just prior to D and C in 100 women presenting with AUB. Smears were reviewed for cytomorphological findings and were correlated with the histopathological findings. These findings were categorized as benign endometrium, endometrial hyperplasia, malignancy and inadequate smears.Results:Age of the patients ranged from 19 to 70 years. In our study, the accuracy in diagnosing benign conditions of endometrium, hyperplasia, and malignancy on aspiration cytology were 93.88%, 96.94% and 96.84%, respectively.Conclusions:Endometrial aspiration is an effective, useful and a minimally invasive procedure. With an experienced cytologist, it can be used routinely for the primary investigation of women with AUB, provided all the points of discrepancies are taken care of.

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Epichloë fungal endophytes of cool season grasses are well-known to produce a range of alkaloids of benefit to the host. Some of these compounds are advantageous to agriculture due to qualities that promote pasture persistence (e.g., the loline class of alkaloids confer insect protection) while others are detrimental to the well-being of grazing livestock. The ergot alkaloids (e.g., ergovaline), produced in ryegrass and tall fescue associations, causes poor animal health in farming regions in many countries around the world and further study is required to improve our knowledge on this class of compounds. Here we present the application of a quantitative LC-MS/MS (liquid chromatography coupled to mass spectrometry) method measuring eight ergot alkaloids (chanoclavine, agroclavine, elymoclavine, lysergol, lysergic acid, ergine, lysergyl-alanine, ergovaline) produced by endophyte infected grasses, to monitor levels in individual tillers from multiple plants of a single cultivar of perennial ryegrass (Lolium perenne cv. “Grasslands Samson”) infected with a common toxic endophyte strain (Epichloë festucae var. lolii). Monitoring the expression in individual tillers allows an estimation of the variability within a plant (between tillers) as well as between plants. The study showed that there is significant variation in the concentration of the ergot alkaloids between tillers of a single plant, at or exceeding the level of variation observed between individual plants of a population. This result emphasizes the fundamental importance of robust experimental design and sampling procedures when alkaloid expression assessment is required and these need to be rigorously tailored to the hypothesis being tested.

Despite the immense capabilities of the date palm, maintaining the fruit’s quality, marketability, and shelf life is still a challenge. This study aimed to assess the synergistic effect of a preharvest spray application of a natural elicitor chitosan, (Ch) 1% alone and in combination with salicylic acid (SA) 2 mM and calcium chloride (Ca) 3%; (Ch,SA, Ca,Ch+Ca, Ch+SA, Ch+SA+Ca), on the quality parameters, storage life, and bioactive compounds content of date fruit from ‘Khasab’ cultivar during cold storage for 60 days. The obtained results revealed that all treatments significantly retard senescence/decay of the fruit compared to the control. Ch+SA treated fruit followed by Ch, and Ch+SA+Ca had the lowest weight loss, color change, and the least decay after 60 days of storage. Ch+Ca, SA, Ca treated fruit had significantly lower levels of total soluble solids and highest total phenolic, tannins, and flavonoids contents compared to the control fruit. Antioxidant activities were found in all treatments, with significantly higher effect in Ch+SA+Ca and Ch+SA compared to the control. Our results provide an evidence for a synergistic effect of elicitors combination to extend the shelf life of date fruit during cold storage by preserving its quality and decreasing senescence/decay and recommend it as a promising strategy.

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Beetroot (Beta vulgaris L.) was subjected to different blanching treatments, that is, T1 (hot-water blanching with cold water immersion), T2 (steam blanching with cold water immersion) and T3 (steam blanching with refrigeration) for obtaining beetroot powder. Fourier transform infrared spectroscopy was used for the evaluation of functional groups in the different blanching treatments. Fresh beetroot and the sample with maximum functional groups, that is, T3 was further used to analyze chemical