Transcriptome analysis of apical meristem enriched bud samples for size dependent flowering commitment in Crocus sativus reveal role of sugar and auxin signalling.

BACKGROUND: Cultivation of Crocus sativus (saffron) faces challenges due to inconsistent flowering patterns and variations in yield. Flowering takes place in a graded way with smaller corms unable to produce flowers. Enhancing the productivity requires a comprehensive understanding of the underlying genetic mechanisms that govern this size-based flowering initiation and commitment. Therefore, samples enriched with non-flowering and flowering apical buds from small (< 6 g) and large (> 14 g) corms were sequenced. METHODS AND RESULTS: Apical bud enriched samples from small and large corms were collected immediately after dormancy break in July. RNA sequencing was performed using Illumina Novaseq 6000 to access the gene expression profiles associated with size dependent flowering. De novo transcriptome assembly and analysis using flowering committed buds from large corms at post-dormancy and their comparison with vegetative shoot primordia from small corms pointed out the major role of starch and sucrose metabolism, Auxin and ABA hormonal regulation. Many genes with known dual responses in flowering development and circadian rhythm like Flowering locus T and Cryptochrome 1 along with a transcript showing homology with small auxin upregulated RNA (SAUR) exhibited induced expression in flowering buds. Thorough prediction of Crocus sativus non-coding RNA repertoire has been carried out for the first time. Enolase was found to be acting as a major hub with protein-protein interaction analysis using Arabidopsis counterparts. CONCLUSION: Transcripts belong to key pathways including phenylpropanoid biosynthesis, hormone signaling and carbon metabolism were found significantly modulated. KEGG assessment and protein-protein interaction analysis confirm the expression data. Findings unravel the genetic determinants driving the size dependent flowering in Crocus sativus.

Effects of Corm Treatment with Cold Plasma and Electromagnetic Field on Growth and Production of Saffron Metabolites in Crocus sativus.

Crocus sativus L. is a widely cultivated traditional plant for obtaining dried red stigmas known as "saffron," the most expensive spice in the world. The response of C. sativus to pre-sowing processing of corms with cold plasma (CP, 3 and 5 min), vacuum (3 min), and electromagnetic field (EMF, 5 min) was assessed to verify how such treatments affect plant performance and the quality and yield of herbal raw materials. The results show that applied physical stressors did not affect the viability of corms but caused stressor-dependent changes in the kinetics of sprouting, growth parameters, leaf trichome density, and secondary metabolite content in stigmas. The effect of CP treatment on plant growth and metabolite content was negative, but all stressors significantly (by 42-74%) increased the number of leaf trichomes. CP3 treatment significantly decreased the length and dry weight of flowers by 43% and 60%, respectively, while EMF treatment increased the length of flowers by 27%. However, longer CP treatment (5 min) delayed germination. Vacuum treatment improved the uniformity of germination by 28% but caused smaller changes in the content of stigma compounds compared with CP and EMF. Twenty-six compounds were identified in total in Crocus stigma samples by the HPLC-DAD method, including 23 crocins, rutin, picrocrocin, and safranal. Processing of Crocus corms with EMF showed the greatest efficiency in increasing the production of secondary metabolites in saffron. EMF increased the content of marker compounds in stigmas (crocin 4: from 8.95 to 431.17 mg/g; crocin 3: from 6.27 to 164.86 mg/g; picrocrocin: from 0.4 to 1.0 mg/g), although the observed effects on growth were neutral or slightly positive. The obtained findings indicate that treatment of C. sativus corms with EMF has the potential application for increasing the quality of saffron by enhancing the amounts of biologically active compounds.

Effect of Shade on Agro-Morphological Parameters and Weed Flora of Saffron (Crocus sativus L.) Cultivation in the Semiarid Zone of Eastern Morocco.

Saffron (Crocus sativus) has been an important medicinal plant since ancient times. This study aimed to seek the optimal light intensity for saffron growth by quantifying the effects of different shade levels on yield, vegetative growth, and weed development in the eastern region of Morocco. The plants were grown for 24 months in full sun (control) and 30%, 50%, and 70% shade. Overall, the results showed that shade positively affected the yield and vegetative growth parameters of saffron plants, with the highest yield (0.61 g/m2) and number of leaves (105 leaves/tuft) recorded when the plants were exposed to light shade (30%). The color of the leaves under the 70% shade levels was dark green. The results from the underground part showed that shade is positively correlated with the weight and diameter of daughter corms where the 70% shade recorded the highest values of weight (65 g) and percentage of large diameter corms (39%). As for weed density, this parameter was significantly affected by shade. The lowest weed density was recorded for the 70% shade treatment. In conclusion, 30% shade is suggested as optimal light irradiation for saffron cultivation.

1H-NMR Metabolic Profiling and Antioxidant Activity of Saffron (Crocus sativus) Cultivated in Lebanon.

Despite the beneficial health properties shown by Lebanese saffron, its qualitative and quantitative composition has never been investigated before. In the present study, NMR spectroscopy, together with antioxidant activity assays, were applied to evaluate the chemical composition of saffron samples of different geographical origins (Lebanon, Italy, Iran, and India) and to categorize the Lebanese saffron for the first time. The distinction between Lebanese saffron and that produced in other countries was attributed to its higher linolenic and linoleic fatty acids, glucose and picrocrocin contents. Moreover, spices produced in three different regions of the Lebanese territory have been clearly differentiated. Saffron cultivated in the Qaa region displayed a high glucose, fatty acids and polyphenols content, whereas Hermel saffron exhibited the largest rate of picrocrocin and glycosylated carotenoids. Finally, samples from Baalbeck showed lower rates for the majority of metabolites. Moreover, Lebanese saffron showed a high antioxidant activity in ABTS and DPPH assays. A low dose of saffron extract (10 µg/mL) inhibited the growth of human lung adenocarcinoma cells, probably due to the high polyphenolic content. This study highlights the quality and peculiarity of Lebanese saffron cultivated in Northern Beqaa district and allows for a good discrimination between spices produced in relatively close territory.

Evaluation of corm origin and climatic conditions on saffron (Crocus sativus L.) yield and quality.

BACKGROUND: Crocus sativus L. is an autumn-flowering geophyte belonging to the Iridaceae family, known for the medicinal and coloring uses of the spice from its dried stigmas. It is cultivated in countries with different pedoclimatic conditions. This paper reports on a two-year research project carried out in the Basilicata region of southern Italy on the question of how to obtain the highest performance from saffron. It considers corms from three different geographical origins - 'Sardinia', 'Abruzzo' and 'Kozani' at three different cultivation sites, namely Castelgrande (40° 46' N, 15° 26' E, 781 m a.s.l.), Genzano di Lucania (40° 50' N, 16° 08' E, 344 m a.s.l.), and Villa d'Agri (40° 22' N, 15° 48' E, 638 m a.s.l.). RESULTS: The highest yields were obtained in the second year by the combination of 'Genzano di L. × Sardinia' and 'Castelgrande × Abruzzo' with 28.1 and 23.9 kg ha-1 of dried stigma, respectively. Saffron quality was determined according to ISO 3632, which classifies the samples into three categories depending on the crocin, picrocrocin, and safranal content. Results highlighted that the highest values for coloring (242.1) and bitterness (97.7) were achieved in Genzano di L. during 2013. Moreover, the crocins were correlated positively with stigma yield and air mean temperature but negatively with safranal. CONCLUSION: This study demonstrated, through principal component analysis (PCA), that the cultivation site with higher air temperature and without excessive rain during the flowering period generated the best stigma yield with high-quality traits. © 2019 Society of Chemical Industry.

Plant growth promoting bacteria associated with corm of Crocus sativus during three growth stages.

Corm, being a unit of reproduction and storage, plays an indispensable role in the life cycle of Crocus sativus (saffron). This study deals with the screening of bacteria isolated from cormosphere (area surrounding the corm sheath) of C. sativus during three growth stages for biofertilizer and biocontrol property. Out of 1000 protease producing bacteria isolated from the cormosphere (soil adhering corm sheath in analogy to rhizosphere), three plant growth promoting (PGP) Bacilli namely Bacillus thuringiensis DC1, B. megaterium VC3 and B. amyloliquefaciens DC8 were selected based on multiple and maximum PGP traits along with corm rot pathogen antagonism. To evaluate in vivo efficacy of the consortia, pot trials were conducted and a significant increase in number and length of both, shoot and root were observed. Further, the disease incidence in test corms was reduced by threefold as compared to control and the number of daughter corms produced in pot trials was also increased. Since corm is a unit of reproduction/propagation, increase in its number is of major significance. This is the first report on isolation and characterization of Plant growth promoting bacteria from any cormosphere.

Unraveling Massive Crocins Transport and Accumulation through Proteome and Microscopy Tools during the Development of Saffron Stigma.

Crocins, the glucosides of crocetin, are present at high concentrations in saffron stigmas and accumulate in the vacuole. However, the biogenesis of the saffron chromoplast, the changes during the development of the stigma and the transport of crocins to the vacuole, are processes that remain poorly understood. We studied the process of chromoplast differentiation in saffron throughout stigma development by means of transmission electron microscopy. Our results provided an overview of a massive transport of crocins to the vacuole in the later developmental stages, when electron dense drops of a much greater size than plastoglobules (here defined "crocinoplast") were observed in the chromoplast, connected to the vacuole with a subsequent transfer of these large globules inside the vacuole. A proteome analysis of chromoplasts from saffron stigma allowed the identification of several well-known plastid proteins and new candidates involved in crocetin metabolism. Furthermore, expressions throughout five developmental stages of candidate genes responsible for carotenoid and apocarotenoid biogenesis, crocins transport to the vacuole and starch metabolism were analyzed. Correlation matrices and networks were exploited to identify a series of transcripts highly associated to crocetin (such as 1-Deoxy-d-xylulose 5-phosphate synthase (DXS), 1-Deoxy-d-xylulose 5-phosphate reductoisomerase (DXR), carotenoid isomerase (CRTISO), Crocetin glucosyltransferase 2 (UGT2), etc.) and crocin (e.g., ζ-carotene desaturase (ZDS) and plastid-lipid-associated proteins (PLAP2)) accumulation; in addition, candidate aldehyde dehydrogenase (ADH) genes were highlighted.

Processed stigmas of Crocus sativus L. imaged by MALDI-based MS.

The processed, i.e. dried under certain conditions, stigmas of Crocus sativus L. are one of the most expensive plant parts used commercially. For the color, aroma and biological activity a very complex mixture of glycolipids termed crocins are responsible. Therefore studying structural composition and distribution in the commercial plant material is of great interest. We showed successfully the application of a MALDI-based mass spectrometric imaging (MSI) approach for stigmas towards different crocin species. MSI opens up the investigation of processed plant materials in various fields allowing studying the processing in detail as well as adulteration attempts (which are quite frequent due to the price of the material). Furthermore, we could demonstrate that a similar number of crocins present in stigmas could be detected by MALDI MSI compared to the classical approach of analyzing the solvent-extract of stigmas by MALDI-MS.

Internet of things (IoT) based saffron cultivation system in greenhouse.

Saffron is the world's most expensive and legendary crop that is widely used in cuisine, drugs, and cosmetics. Therefore, the demand for saffron is increasing globally day by day. Despite its massive demand the cultivation of saffron has dramatically decreased and grown in only a few countries. Saffron is an environment-sensitive crop that is affected by various factors including rapid change in climate, light intensity, pH level, soil moisture, salinity level, and inappropriate cultivation techniques. It is not possible to control many of these environmental factors in traditional farming. Although, many innovative technologies like Artificial Intelligence and Internet of Things (IoT) have been used to enhance the growth of saffron still, there is a dire need for a system that can overcome primary issues related to saffron growth. In this research, we have proposed an IoT-based system for the greenhouse to control the numerous agronomical variables such as corm size, temperature, humidity, pH level, soil moisture, salinity, and water availability. The proposed architecture monitors and controls environmental factors automatically and sends real-time data from the greenhouse to the microcontroller. The sensed values of various agronomical variables are compared with threshold values and saved at cloud for sending to the farm owner for efficient management. The experiment results reveal that the proposed system is capable to maximize saffron production in the greenhouse by controlling environmental factors as per crop needs.

Revealing the dynamics of saffron growth: Optimizing corm size and planting depth for increased yield synergies.

Saffron, the "golden spice" derived from Crocus sativus L., is renowned for its richness in secondary metabolites such as crocin and safranal, contributing to its unique properties. Facing challenges like decreasing global production, optimizing cultivation techniques becomes imperative for enhanced yields. Although the impact of factors like planting density, planting depth, spacing, and corm size on saffron growth has been studied, the interaction between corm size and planting depth remains underexplored. This study systematically investigates the interactive effects of corm size and planting depth on saffron growth and yield, providing evidence-based guidelines for optimizing cultivation. A factorial experiment, employing a completely randomized design, was conducted to assess the influence of corm size (05-10g, 10.1-15g, 15.1-20g) and planting depth (10cm, 15cm, 20cm) on saffron yield. Uniform-sized corms were obtained, and a suitable soil mixture was prepared for cultivation. Morphological and agronomic parameters were measured, and statistical analyses were performed using ANOVA and Tukey's HSD test. The study revealed that planting depth significantly affected saffron emergence. The corms sown under 15cm depth showed 100% emergence regardless of corm size (either 05-10g, 10.1-15g, 15.1-20g) followed by 10cm depth corms. Corm dry weight exhibited a complex interaction, where larger corms benefited from deeper planting, while intermediate-sized corms thrived at shallower depths. Similar patterns were observed in shoot fresh weight and dry weight. Specifically, the largest corm size (t3, 15.1-20g) produced the greatest fresh-weight biomass at the deepest planting depth of 20cm (T3), while intermediate-sized corms (t2, 10.1-15g) were superior at the shallowest 10cm depth (T1). The total plant biomass demonstrated that larger corms excelled in deeper planting, while intermediate-sized corms were optimal at moderate depths. This research highlights the intricate interplay between corm size and planting depth in influencing saffron growth. Larger corms generally promote higher biomass, but the interaction with planting depth is crucial. Understanding these dynamics can aid farmers in tailoring cultivation practices for optimal saffron yields. The study emphasizes the need for a coordinated approach to corm selection and depth placement, providing valuable insights for sustainable saffron production and economic growth.

Plant growth promoting bacteria from Crocus sativus rhizosphere.

Present study deals with the isolation of rhizobacteria and selection of plant growth promoting bacteria from Crocus sativus (Saffron) rhizosphere during its flowering period (October-November). Bacterial load was compared between rhizosphere and bulk soil by counting CFU/gm of roots and soil respectively, and was found to be ~40 times more in rhizosphere. In total 100 bacterial isolates were selected randomly from rhizosphere and bulk soil (50 each) and screened for in-vitro and in vivo plant growth promoting properties. The randomly isolated bacteria were identified by microscopy, biochemical tests and sequence homology of V1-V3 region of 16S rRNA gene. Polyphasic identification categorized Saffron rhizobacteria and bulk soil bacteria into sixteen different bacterial species with Bacillus aryabhattai (WRF5-rhizosphere; WBF3, WBF4A and WBF4B-bulk soil) common to both rhizosphere as well as bulk soil. Pseudomonas sp. in rhizosphere and Bacillus and Brevibacterium sp. in the bulk soil were the predominant genera respectively. The isolated rhizobacteria were screened for plant growth promotion activity like phosphate solubilization, siderophore and indole acetic acid production. 50 % produced siderophore and 33 % were able to solubilize phosphate whereas all the rhizobacterial isolates produced indole acetic acid. The six potential PGPR showing in vitro activities were used in pot trial to check their efficacy in vivo. These bacteria consortia demonstrated in vivo PGP activity and can be used as PGPR in Saffron as biofertilizers.This is the first report on the isolation of rhizobacteria from the Saffron rhizosphere, screening for plant growth promoting bacteria and their effect on the growth of Saffron plant.

Developmental and stress regulation of gene expression for a 9-cis-epoxycarotenoid dioxygenase, CstNCED, isolated from Crocus sativus stigmas.

Oxidative cleavage of cis-epoxycarotenoids by 9-cis-epoxycarotenoid dioxygenase (NCED) is the critical step in the regulation of abscisic acid (ABA) synthesis in higher plants. ABA has been associated with dormancy and flower senescence, while also regulating plant adaptive responses to various environmental stresses. An NCED gene, CstNCED, was cloned from Crocus sativus stigmas. The deduced amino acid sequence of the CstNCED protein shared high identity with other monocot NCEDs, and was closely related to the liliopsida enzymes. At the N-terminus of CstNCED a chloroplast transit peptide sequence is located. However, its expression in chloroplast-free tissues suggested localization in other plastid types. The relationship between expression of CstNCED and the endogenous ABA level was investigated in the stigma and corms, where it was developmentally regulated. The senescence of the unpollinated stigma is preceded by an increase in ABA levels and CstNCED expression. In corms, a correlation was observed between CstNCED expression and dormancy. Furthermore, CstNCED expression was correlated with the presence of zeaxanthin in the dormant corms. When detached C. sativus leaves and stigmas were water and salt stressed, increases in CstNCED mRNA were observed. The results provided evidence of the involvement of CstNCED in the regulation of ABA-associated processes such as flower senescence and corm dormancy in monocotyledonous saffron.

Field evaluation of PGP Bacillus sp. strain D5 native to Crocus sativus, in traditional and non traditional areas, and mining of PGP genes from its genome.

Native Bacillus sp. strain D5 coded as (Bar D5) has been isolated from the saffron corm that showed plant growth promotion (PGP) properties and also inhibits the growth of corm rot causing Fusarium oxysporum R1 (Fox R1) in-vitro. Bar D5 was more efficient PGP bacterium in comparison to earlier reported native bio-formulations by our group. Pot assays and field evaluation of Bar D5 confirmed its in-vivo efficacy for PGP traits and biocontrol activity as well. Pot trials were followed by field trials at traditional (Kishtwar) and non-traditional (R.S Pura) saffron cultivation areas in Jammu and Kashmir. At both places, Bar D5 bio-formulation treatment led to the increase in root number & length, shoot number & length, flower number and number & weight of daughter corms. Additionally, it also decreased the corm rot disease incidence significantly. Priming of corms with bio-formulation resulted in the reduction of pathogenic fungal load by three fold at the depth of corm sowing from ground level. The shelf life/viability of Bar D5 based bio-formulation was found to be 52% (viable spores) for one year at room temperature. Draft genome sequence of Bar D5 revealed the presence of genes necessary for PGP and biocontrol activity. Further, confirmation of gene sequences and annotation was done by amplification, re-sequencing and mapping of PGP and biocontrol genes on draft genome. Bar D5 based bio-formulation can be provided to companies/researchers interested in saffron cultivation or bio-formulation production for commercial exploitation, since saffron is grown as revenue crop across continents. The present study bridges the gap between genomics and its field application.

Blue Light Improves Photosynthetic Performance and Biomass Partitioning toward Harvestable Organs in Saffron (Crocus sativus L.).

Saffron is a valuable plant and one of the most expensive spices worldwide. Nowadays, there is a tendency to produce this crop in indoor plant production systems. However, the production of saffron is restricted by the need for the reproduction of high-quality corms. In this study, we investigated the effect of different ratios of red (R) and blue (B) light spectra (including 100% B (monochromatic B), 75%, 50%, 40%, 25% B, and 0% B (monochromatic R) on the photosynthetic performance and biomass partitioning as well as morphological and biochemical characteristics of saffron. The growth of flower, root, and corm was improved by increasing the proportion of B to R light. B-grown plants were characterized by the highest photosynthetic functionality with efficient electron transport and lower energy dissipation when compared to R-grown plants. B light directed biomass toward the corms and floral organs, while R light directed it toward the leaves. In saffron, the weight of a daughter corm is of great importance since it determines the yield of the next year. As the ratio of B to R light increased, the daughter corms also became heavier, at the cost of reducing their number, though increasing the proportion of B-enhanced antioxidant capacity as well as the activity of ascorbate peroxidase and catalase while superoxide dismutase activity was enhanced in R-grown plants. In conclusion, B light increased the production of high-quality daughter corms and altered biomass partitioning towards harvestable organs (corms and flowers) in saffron plants.

Differential interaction of Or proteins with the PSY enzymes in saffron.

Colored apocarotenoids accumulate at high concentrations in few plant species, where display a role in attraction of pollinators and seed dispersers. Among these apocarotenoids, crocins accumulate at high concentrations in the stigma of saffron and are responsible for the organoleptic and medicinal properties of this spice. Phytoene synthase and Orange protein are key for carotenoid biosynthesis and accumulation. We previously isolated four phytoene synthase genes from saffron with differential roles in carotenoid and apocarotenoid biosynthesis. However, the implications of Orange genes in the regulation of apocarotenoid accumulation are unknown. Here, we have identified two Orange genes from saffron, with different expression patterns. CsOr-a was mainly expressed in vegetative tissues and was induced by light and repressed by heat stress. Both CsOr-a and CsOr-b were expressed in stigmas but showed a different profile during the development of this tissue. The interactions of CsOr-a and CsOr-b were tested with all the four phytoene synthase proteins from saffron and with CsCCD2. None interactions were detected with CCD2 neither with the phytoene synthase 2, involved in apocarotenoid biosynthesis in saffron. The obtained results provide evidence of different mechanisms regulating the phytoene synthase enzymes in saffron by Orange for carotenoid and apocarotenoid accumulation in saffron.

Low temperature maximizes growth of Crocus vernus (L.) Hill via changes in carbon partitioning and corm development.

In Crocus vernus, a spring bulbous species, prolonged growth at low temperatures results in the development of larger perennial organs and delayed foliar senescence. Because corm growth is known to stop before the first visual sign of leaf senescence, it is clear that factors other than leaf duration alone determine final corm size. The aim of this study was to determine whether reduced growth at higher temperatures was due to decreased carbon import to the corm or to changes in the partitioning of this carbon once it had reached the corm. Plants were grown under two temperature regimes and the amount of carbon fixed, transported, and converted into a storable form in the corm, as well as the partitioning into soluble carbohydrates, starch, and the cell wall, were monitored during the growth cycle. The reduced growth at higher temperature could not be explained by a restriction in carbon supply or by a reduced ability to convert the carbon into starch. However, under the higher temperature regime, the plant allocated more carbon to cell wall material, and the amount of glucose within the corm declined earlier in the season. Hexose to sucrose ratios might control the duration of corm growth in C. vernus by influencing the timing of the cell division, elongation, and maturation phases. It is suggested that it is this shift in carbon partitioning, not limited carbon supply or leaf duration, which is responsible for the smaller final biomass of the corm at higher temperatures.

Hormone and microorganism treatments in the cultivation of saffron (Crocus sativus L.) plants.

The difficult cultivation of the saffron plant (Crocus Sativus L.) make the spice of the same name made from its dried stigmas very valuable. It is estimated that some 75,000 blossoms or 225,000 hand-picked stigmas are required to make a single pound of saffron, which explains why it is the world's most expensive spice. The aim of this study was to identify ways of increasing the fertility and production of saffron. For this purpose, the treatment of saffron bulbs with a synthetic growth hormone--a mixture of Polystimulins A6 and K--and two different microorganism based materials--biohumus or vermicompost and Effective Microorganisms (EM)--in four different ways (hormone alone, biohumus alone, EM alone and EM+biohumus) was investigated to determine whether these treatments have any statistically meaningful effects on corms and stigmas. It has been shown that EM + biohumus was the most effective choice for improved saffron cultivation.

Transcriptome analysis in tissue sectors with contrasting crocins accumulation provides novel insights into apocarotenoid biosynthesis and regulation during chromoplast biogenesis.

Crocins, the red soluble apocarotenoids of saffron, accumulate in the flowers of Crocus species in a developmental and tissue-specific manner. In Crocus sieberi, crocins accumulate in stigmas but also in a distinct yellow tepal sector, which we demonstrate contains chromoplast converted from amyloplasts. Secondary metabolites were analysed by LC-DAD-HRMS, revealing the progressive accumulation of crocetin and crocins in the yellow sector, which were also localized in situ by Raman microspectroscopy. To understand the underlying mechanisms of crocin biosynthesis, we sequenced the C. sieberi tepal transcriptome of two differentially pigmented sectors (yellow and white) at two developmental stages (6 and 8) by Illumina sequencing. A total of 154 million high-quality reads were generated and assembled into 248,099 transcripts. Differentially expressed gene analysis resulted in the identification of several potential candidate genes involved in crocin metabolism and regulation. The results provide a first profile of the molecular events related to the dynamics of crocetin and crocin accumulation during tepal development, and present new information concerning apocarotenoid biosynthesis regulators and their accumulation in Crocus. Further, reveals genes that were previously unknown to affect crocin formation, which could be used to improve crocin accumulation in Crocus plants and the commercial quality of saffron spice.

Kinetics properties of guaiacol peroxidase activity in Crocus sativus L. corm during rooting.

BACKGROUND: Guaiacol peroxidases (GP) are haem-containing enzymes participating in many physiological processes in plants. The expression pattern of these enzymes is organ-specific and developmentally regulated. METHODS: The presence of GP activity in extract samples, prepared from Crocus sativus L. corms that were either dormant or rooting for 3, 6 and 10 days, was investigated. RESULTS: Kinetic studies revealed a significant similarity among GP activities detectable in the corm at different stages of development: in all extract samples, the activity was maximal at pH 7.5 and after preincubation at 30-40 degrees C. When guaiacol was used as the varying substrate, Michaelis-Menten kinetics behavior was observed in all extract samples and resulted in similar KM values; catalytic efficiencies were also very similar. The corm GP activity was inhibited by cyanide, azide and ascorbate. The GP activities from different extract samples had the same sensitivities to azide, cyanide and ascorbate and the type of inhibition by azide and cyanide was competitive and uncompetitive, respectively, while ascorbate inhibited the GP activity non-competitively. Corm extract samples from different stages of rooting similarly responded to temperature treatment and a biphasic Arrhenius plot resulted for each extract sample studied. When dormant, 3-, 6- and 10-days-rooting corm extracts were submitted to non-denaturing polyacrylamide gel electrophoresis, the GP-specific activity staining revealed one band on the gel, with the same migrating distances. CONCLUSION: This finding in combination with kinetic studies demonstrated that at least one form of GP, with an apparent molecular weight of 68 kDa, was expressed during development of Crocus sativus L. corm.

Life cycle environmental impacts of saffron production in Iran.

Saffron is regarded as an important crop in Iranian agricultural economics that needs to be investigated to produce the environmentally friendlier product. In the present study, saffron production as an important agricultural production system in Iran was evaluated thoroughly from an environmental point of view. Data were collected from saffron farmers in Southern Khorasan province of the country with face-to-face questionnaire method during cropping period of 2013-2014. The system boundary was considered from the production of raw input materials to the harvested saffron. In order to identify the main hotspot during cultivation, impact categories were considered using CML methodology and cumulative exergy demand (CExD) indicator. Based on the results, chemical fertilizers N and P were the main hotspots in abiotic depletion (AD), acidification (AC), global warming (GWP), and photochemical oxidation (PO) impact categories, while diesel fuel was the main hotspot in fresh water aquatic ecotoxicity (FE), marine aquatic ecotoxicity (ME), and terrestrial ecotoxicity (TE). Direct emission from diesel fuel combustion (saffron) was the main hotspot only in human toxicity (HT). The small farms had the highest amounts of AD, AC, EP, GWP, and PO indicators in comparison with the medium and large farms. Total CExD indicator for production of 1 kg saffron was 1894.23 MJ eq. Totally, large farms (bigger than 1 ha) had better environmental performance considering all the impact categories.