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Nasheeman Ashraf – One of the best experts on this subject based on the ideXlab platform.

  • Identification, phylogenetic analysis and expression profiling of ABC transporter family of Crocus sativus L: A step towards understanding Apocarotenoid transport
    Plant Gene, 2018
    Co-Authors: Tabasum Mohiuddin, Shoib Ahmad Baba, Nasheeman Ashraf

    Abstract:

    Abstract Crocus sativus has recently become focus of research because it is the only plant species which produces Apocarotenoids like crocins, picrocrocin and safranal in significant amounts. The compounds are synthesized only in stigma part of the flower, the dried form of which forms commercial saffron. These compounds impart organoleptic properties to saffron making it world’s costliest spice. The Crocus Apocarotenoids are synthesized in plastoglobules and are later transported to vacuole for storage. They also undergo long distance transport from the site of their synthesis to that of their action. Till so far, very less information is available regarding the mechanism of their transport. In this context, the present study involves identification, phylogenetic analysis and expression profiling of ATP-binding cassette (ABC) genes from Crocus with the aim of gaining information about the genes involved in Apocarotenoid transport. ABC proteins represent one of the largest protein families in plants and are known to play role in a number of biological processes. Here we identified 77 ABC transporter genes from Crocus transcriptome. Phylogenetic analysis of Crocus ABC genes with those of Arabidopsis divided them into 8 subfamilies among which ABCB and ABCG were most predominant. Expression analysis of one member from each subfamily was performed which demonstrated that the genes showed tissue specific expression. Further ABC genes belonging to different subfamilies were induced in response to different stress and hormone treatments suggesting their role in diverse biological processes. These results would facilitate further investigation into their involvement in Crocus Apocarotenoid metabolism and transport.

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  • mortierella alpina cs10e4 an oleaginous fungal endophyte of crocus sativus l enhances Apocarotenoid biosynthesis and stress tolerance in the host plant
    Scientific Reports, 2017
    Co-Authors: Zahoor A Wani, Nasheeman Ashraf, Amit Kumar, Phalisteen Sultan, Kushal Bindu, Syed Riyazulhassan

    Abstract:

    Crocus sativus is the only plant species which produces Apocarotenoids like crocin, picrocrocin and safranal in significant amounts. These compounds impart organoleptic properties to saffron (dried stigmas of Crocus flower) making it world’s costliest spice. Crocus Apocarotenoids have tremendous medicinal properties as well. Effect of endophytes on Crocus Apocarotenoid production and the molecular mechanism involved has not been reported so far. Here we studied the effect of an oleaginous fungal endophyte, Mortierella alpina CS10E4 on Crocus growth, Apocarotenoid metabolism and tolerance to corm rot disease. The results demonstrated that there was a significant improvement in many morphological and physiological traits in endophyte treated Crocus plants including total biomass and size of corms, stigma biomass, number of apical sprouting buds, and number of adventitious roots. The endophyte also shifted metabolic flux towards enhanced production of Apocarotenoids by modulating the expression of key pathway genes. Further, M. alpina CS10E4 enhanced tolerance to corm rot disease by releasing arachidonic acid which acts as conserved defense signal and induces jasmonic acid production in endophyte treated Crocus corms. This is first report on effect of a fungal endophyte on Crocus Apocarotenoid metabolism and stress tolerance.

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  • Transcriptome wide identification, phylogenetic analysis, and expression profiling of zinc-finger transcription factors from Crocus sativus L.
    Molecular Genetics and Genomics, 2017
    Co-Authors: Aubid Hussain Malik, Nasheeman Ashraf

    Abstract:

    Crocus sativus belongs to Iridaceae family and is the only plant species which produces Apocarotenoids like crocin, picrocrocin, and safranal in significant quantities. Besides their organoleptic properties, Crocus Apocarotenoids have been found to possess remarkable pharmacological potential. Although Apocarotenoid biosynthetic pathway has been worked out to a great degree, but the mechanism that regulates the tissue and developmental stage-specific production of Crocus Apocarotenoids is not known. To identify the genes regulating Apocarotenoid biosynthesis in Crocus , transcriptome wide identification of zinc-finger transcription factors was undertaken. 81 zinc-finger transcription factors were identified which grouped into eight subfamilies. C2H2, C3H, and AN20/AN1 were the major subfamilies with 29, 20, and 14 members, respectively. Expression profiling revealed CsSAP09 as a potential candidate for regulation of Apocarotenoid biosynthesis. CsSAP09 was found to be highly expressed in stigma at anthesis stage corroborating with the accumulation pattern of Apocarotenoids. CsSAP09 was nuclear localized and activated reporter gene transcription in yeast. It was highly induced in response to oxidative, salt and dehydration stresses, ABA and methyl jasmonate. Furthermore, upstream region of CsSAP09 was found to contain stress and light responsive elements. To our knowledge, this is the first report on the study of a gene family in C. sativus and may provide basic insights into the putative role of zinc finger genes . It may also serve as a valuable resource for functional characterization of these genes aimed towards unraveling their role in regulation of Apocarotenoid biosynthesis.

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Oussama Ahrazem – One of the best experts on this subject based on the ideXlab platform.

  • efficient production of saffron crocins and picrocrocin in nicotiana benthamiana using a virus driven system
    Metabolic Engineering, 2020
    Co-Authors: Maricarmen Marti, Gianfranco Diretto, Lourdes Gomezgomez, Oussama Ahrazem, Veronica Aragones, Sarah Frusciante, José-antonio Daròs

    Abstract:

    Abstract Crocins and picrocrocin are glycosylated Apocarotenoids responsible, respectively, for the color and the unique taste of the saffron spice, known as red gold due to its high price. Several studies have also shown the health-promoting properties of these compounds. However, their high costs hamper the wide use of these metabolites in the pharmaceutical sector. We have developed a virus-driven system to produce remarkable amounts of crocins and picrocrocin in adult Nicotiana benthamiana plants in only two weeks. The system consists of viral clones derived from tobacco etch potyvirus that express specific carotenoid cleavage dioxygenase (CCD) enzymes from Crocus sativus and Buddleja davidii. Metabolic analyses of infected tissues demonstrated that the sole virus-driven expression of C. sativus CsCCD2L or B. davidii BdCCD4.1 resulted in the production of crocins, picrocrocin and safranal. Using the recombinant virus that expressed CsCCD2L, accumulations of 0.2% of crocins and 0.8% of picrocrocin in leaf dry weight were reached in only two weeks. In an attempt to improve Apocarotenoid content in N. benthamiana, co-expression of CsCCD2L with other carotenogenic enzymes, such as Pantoea ananatis phytoene synthase (PaCrtB) and saffron β-carotene hydroxylase 2 (BCH2), was performed using the same viral system. This combinatorial approach led to an additional crocin increase up to 0.35% in leaves in which CsCCD2L and PaCrtB were co-expressed. Considering that saffron Apocarotenoids are costly harvested from flower stigma once a year, and that Buddleja spp. flowers accumulate lower amounts, this system may be an attractive alternative for the sustainable production of these appreciated metabolites.

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  • ugt709g1 a novel uridine diphosphate glycosyltransferase involved in the biosynthesis of picrocrocin the precursor of safranal in saffron crocus sativus
    New Phytologist, 2019
    Co-Authors: Gianfranco Diretto, Oussama Ahrazem, Angela Rubiomoraga, Alessia Fiore, Filippo Sevi, Javier Argandoña, Lourdes Gomezgomez

    Abstract:

    : Saffron, a spice derived from the dried red stigmas of Crocus sativus, is one of the oldest natural food additives. The flowers have long red stigmas, which store significant quantities of the glycosylated Apocarotenoids crocins and picrocrocin. The Apocarotenoid biosynthetic pathway in saffron starts with the oxidative cleavage of zeaxanthin, from which crocins and picrocrocin are derived. In the processed stigmas, picrocrocin is converted to safranal, giving saffron its typical aroma. By a targeted search for differentially expressed uridine diphosphate glycosyltransferases (UGTs) in Crocus transcriptomes, a novel Apocarotenoid glucosyltransferase (UGT709G1) from saffron was identified. Biochemical analyses revealed that UGT709G1 showed a high catalytic efficiency toward 2,6,6-trimethyl-4-hydroxy-1-carboxaldehyde-1-cyclohexene (HTCC), making it suited for the biosynthesis of picrocrocin, the precursor of safranal. The role of UGT709G1 in picrocrocin/safranal biosynthesis was supported by the absence or presence of gene expression in a screening for HTCC and picrocrocin production in different Crocus species and by a combined transient expression assay with CsCCD2L in Nicotiana benthamiana leaves. The identification of UGT709G1 completes one of the most highly valued specialized metabolic biosynthetic pathways in plants and provides novel perspectives on the industrial production of picrocrocin to be used as a flavor additive or as a pharmacological constituent.

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  • Multi-species transcriptome analyses for the regulation of crocins biosynthesis in Crocus
    BMC Genomics, 2019
    Co-Authors: Oussama Ahrazem, Alessia Fiore, Javier Argandoña, Andrea Rujas, Angela Rubio-moraga, Raquel Castillo, Lourdes Gómez-gómez

    Abstract:

    Crocins are soluble Apocarotenoids that mainly accumulate in the stigma tissue of Crocus sativus and provide the characteristic red color to saffron spice, in addition to being responsible for many of the medicinal properties of saffron. Crocin biosynthesis and accumulation in saffron is developmentally controlled, and the concentration of crocins increases as the stigma develops. Until now, little has been known about the molecular mechanisms governing crocin biosynthesis and accumulation. This study aimed to identify the first set of gene regulatory processes implicated in Apocarotenoid biosynthesis and accumulation. A large-scale crocin-mediated RNA-seq analysis was performed on saffron and two other Crocus species at two early developmental stages coincident with the initiation of crocin biosynthesis and accumulation. Pairwise comparison of unigene abundance among the samples identified potential regulatory transcription factors (TFs) involved in crocin biosynthesis and accumulation. We found a total of 131 (up- and downregulated) TFs representing a broad range of TF families in the analyzed transcriptomes; by comparison with the transcriptomes from the same developmental stages from other Crocus species, a total of 11 TF were selected as candidate regulators controlling crocin biosynthesis and accumulation. Our study generated gene expression profiles of stigmas at two key developmental stages for Apocarotenoid accumulation in three different Crocus species. Differential gene expression analyses allowed the identification of transcription factors that provide evidence of environmental and developmental control of the Apocarotenoid biosynthetic pathway at the molecular level.

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Nora M Obrien – One of the best experts on this subject based on the ideXlab platform.

  • cellular transport and bioactivity of a major saffron Apocarotenoid picrocrocin 4 β d glucopyranosyloxy 2 6 6 trimethyl 1 cyclohexene 1 carboxaldehyde
    Journal of Agricultural and Food Chemistry, 2015
    Co-Authors: Anastasia Kyriakoudi, Yvonne C Ocallaghan, Karen Galvin, Maria Z. Tsimidou, Nora M Obrien

    Abstract:

    The cellular transport and bioactivity of the second major saffron Apocarotenoid, picrocrocin, was examined in parallel to that of the major group, crocetin sugar esters, in aqueous extracts. The transport of pure picrocrocin was investigated in comparison to that of other saffron Apocarotenoids, trans-crocetin (di-β-d-gentiobiosyl) ester and crocetin using the Caco-2 cell model coupled with an in vitro digestion procedure. RP-HPLC-DAD was employed to quantify the bioaccessible and bioavailable amounts of individual Apocarotenoids. Picrocrocin and crocetin sugar esters though highly bioaccessible (75% and 60%, respectively) were transported at minute quantities (0.2% and 0.5%, respectively; 10-fold lower than crocetin). Picrocrocin did not protect against oxidant-induced DNA damage in U937, human monocytic blood cells at the concentration investigated, however, it reduced the proliferation of human adenocarcinoma and hepatocarcinoma cells. Our findings may be useful for the requirements of food legislatio…

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