不同产地红花的挥发油成分

不同产地红花的挥发油成分郭美丽张汉明张芝玉苏中武（第二军医大学药学院,上海２００４３３）Ｔｈｅｃｏｎｓｔｉｔｕｅｎｔｓｏｆｅｓｅｎｔｉａｌｏｉｌｆｒｏｍｓａｆｆｌｏｗｅｒ（ＣａｒｔｈａｍｕｓｔｉｎｃｔｏｒｉｕｓＬ．）ｉｎｄｉｆｆｅｒｅｎｔｌｏｃａｌｉ...     

EMS-induced cytomictic variability in safflower (Carthamus tinctorius L.)

Seeds of safflower (Carthamus tinctorius L.) were subjected to three treatment durations (3h, 5h and 7h) of 0.5 % Ethyl Methane Sulphonate (EMS). Microsporogenesis was carried out in the control as well as in the treated materials. EMS treated plants showed interesting feature of partial inter-meiocyte chromatin migration through channel formation, beak formation or direct cell fusion. Another interesting feature noticed during the study was the fusion among tetrads due to wall dissolution. The phenomenon of cytomixis was recorded at nearly all the stages of microsporogenesis connecting from a few to several meiocytes. Other abnormalities such as laggards, precocious movement, bridge and non-disjunction of chromosomes were also recorded but in very low frequencies. The phenomenon of cytomixis increased along with the increase in treatment duration of EMS. Cells with these types of cytomictic disturbances may probably result in uneven formation of gametes or zygote, heterogenous sized pollen grains or even loss of fertility in future.     

Improvements in rooting regenerated safflower (Carthamus tinctorius L.) shoots

A continuing obstacle for regenerating safflower (Carthamus tinctorius L.) plants from cultured explants or callus has been a reliable method for rooting shoots. For shoots directly regenerated from primary explants, 76% of shoots rooted after a 7-d exposure to 10 mg/1 indole-3-butyric acid. Auxin source, concentration or exposure time did not greatly affect root formation or morphology, but strongly affected callus production. Shoots infected with Agrobacterium rhizogenes produced massive numbers of fibrous roots, but shoots did not elongate or survive transfer to soil. Shoot hyperhydricity symptoms were reduced by including 1 g/1 activated charcoal in rooting media. The optimal protocol for inducing root formation consisted of a 7-d exposure to 10 mg/l indole-3-butyric acid in root induction media, followed by incubation in media containing 15 g/l sucrose and 1 g/1 activated charcoal for 21 d.Abbreviations IBA indole-3-butyric acid - MS Murashige and Skoog (1962) medium - NAA anaphthalene acetic acid - POP 2,3,5-trichloro--phenoxypropionic acid     

Association-dissociation and denaturation-renaturation of high-molecular-weight protein: carmin from safflower seed (Carthamus tinctorius L.) in alkaline solution

The effect of alkalinepH on the association, dissociation, and denaturation of carmin, the high-molecular-weight protein from safflower seed was investigated in thepH range 7–12, using various biophysical techniques. The results indicate that the multimeric protein carmin dissociates atpH 8.0 where denaturation has not set in. The association-dissociation of the protein can be represented schematically as 11S 7S 4S 2S. AbovepH 10, the protein undergoes simultaneous dissociation and denaturation. The denaturation process appears to be complete at pH 12.5. The protein undergoes conformational change and covalent modifications and cleavage during the denaturation process. A reversibility study shows that the process of dissociation is reversible to a large extent, whereas denaturation appears to be irreversible. These results are discussed in terms of association-dissociation, denaturation and alkaline-catalyzed covalent modifications and cleavage of seed proteins.     

Inheritance of flower color and spininess in safflower (Carthamus tinctorius L.)

Safflower (Carthamus tinctorius L.) flowers are used for coloring and flavoring food and also as fresh-cut and dried flowers. The most important characteristics which contribute to the ornamental value of safflower are flower color and spinelessness. The objective of this study was to determine the inheritance mode and the number of genes controlling spininess and flower color in some Iranian genotypes of safflower. The results indicated that the existence of spines on the leaves and bracts of safflower is controlled by a single dominant gene in which the spiny phenotype was completely dominant to spineless. In some crosses, flower color was controlled by two epistatic loci each with two alleles, resulting in a ratio of 13:3 in the segregating F2 population for plants with orange and yellow flowers. Also, other mechanisms of genetic control, such as duplicate dominance and duplicate recessive types of epistasis, were observed for flower color in other crosses that led to ratios of 7:9 and 15:1 for plants with orange and yellow flowers, respectively. The results suggest that for ornamental use or in the food dying industry, genotypes with orange or yellow flowers and without spines on the leaves and bracts can be produced.     

Screen of safflower (Carthamus tinctorius L.) genotypes against Phytophthora drechsleri and Fusarium solani,the causal agents of root rot disease

Safflower (Carthamus tinctorius L.) plants were affected by a severe root rot disease caused by Phytophthora drechsleri and Fusarium solani in Isfahan province of Iran during 2005–2007. Disease incidence was more than 30% in severely infected fields. Twenty-one safflower genotypes, including six local cultivars and 15 internal pure lines were evaluated for their resistance to root rot disease in laboratory and greenhouse conditions. Safflower seedlings were evaluated for lesion length on infected roots in laboratory, as well percentage of live seedlings in greenhouse. The results indicated a high negative correlation between lesion length on roots and percentage of live seedlings. The most resistant and susceptible genotypes to P. drechsleri were identified as pure line Karaj row 12 (KW12) and cultivar Koseh with lesion lengths of 10.01 and 15.51?mm on roots and 45.60 and 18.00% live seedlings, respectively. The most resistant genotype to F. solani was identified as pure line KW11 with a lesion length of 9.31?mm on roots as well 62.80% live seedlings. The most susceptible genotypes were identified as cultivar Koseh and pure lines KW2 and KW3 with lesion lengths of 13.29, 12.72 and 12.13?mm on roots and 25.60, 28.40 and 28.40% live seedlings, respectively. The most resistant genotypes to both P. drechsleri and F. solani were identified as pure lines KW15 and KW9 with a 55.40% live seedlings. The most susceptible genotypes were cultivars Koseh, Goldasht and pure lines KW6, KW3 and KW2 with 35.40, 35.40, 35.40, 37.60 and 37.60% live seedlings in greenhouse, respectively.     

Restriction endonuclease cleavage site map of safflower (Carthamus tinctorius L.) chloroplast DNA

Summary The restriction endonucleases SalI, PstI, KpnI and HindIII have been used to construct a physical map of safflower (Carthamus tinctorius L.) chloroplast DNA. This was accomplished by hybridizing Southern blots of single and double digested chloroplast DNA with ³²P-dCTP nick-translated SalI, KpnI and HindIII probes which were individually isolated from agarose gels. The chloroplast DNA was found to be circular and to contain approximately 151 kbp. In common with many other higher plant chloroplast DNAs a sequence of about 25 kbp is repeated in an inverted orientation. The small and large single copy regions separating the two repeated segments contain about 20 kbp and 81 kbp, respectively. The rRNA structural genes were also mapped by Southern blot hybridization and are co-linear with several other plant species.     

Expression and recovery of biologically active recombinant Apolipoprotein AIMilano from transgenic safflower (Carthamus tinctorius) seeds

Apolipoprotein AI Milano (ApoAI_Milano) was expressed as a fusion protein in transgenic safflower seeds. High levels of expression corresponding to 7 g of ApoAI_Milano per kilogram of seed have been identified in a line selected for commercialization. The ApoAI_Milano fusion protein was extracted from seed using an oilbody‐based process and matured in vitro prior to final purification. This yielded a Des‐1,2‐ApoAI_Milano product which was confirmed by biochemical characterization including immunoreactivity against ApoAI antibodies, isoelectric point, N‐terminal sequencing and electrospray mass spectrometry. Purified Des‐1,2‐ApoAI_Milano readily associated with dimyristoylphosphatidylcholine in clearance assays comparable to Human ApoAI. Its biological activity was assessed by cholesterol efflux assays using Des‐1,2‐ApoAI_Milano:1‐palmitoyl‐2‐oleoyl phosphatidylcholine complexes in vitro and in vivo. This study has established that high levels of biologically functional ApoAI_Milano can be produced using a plant‐based expression system.     

Central role of salicylic acid in resistance of safflower (Carthamus tinctorius L.) against salinity

The effects of salicylic acid (SA) on growth parameters and enzyme activities were investigated in salt-stressed safflower (Carthamus tinctorius L.). Twenty-five days after sowing, seedlings were treated with NaCl (0, 100, and 200?mM) and SA (1?mM), and were harvested at 21 days after treatments. Results showed that some growth parameters decreased under salinity, while malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) content, phenolic compounds, and some enzyme activities increased. SA application increased some growth parameters, MDA and H₂O₂ content, and enzyme activities except catalase (CAT), which was different from the other enzymes and SA significantly reduced CAT activity in plants. These results suggest that SA-induced tolerance to salinity may be related to regulation of antioxidative responses and H₂O₂ level. Our study suggested that the resistant safflower can direct reactive oxygen species from a threat to an opportunity by using SA. Therefore, exogenous application of SA played this role through regulation of the antioxidant system.     

Polyacetylenes from Immature Seeds of Safflower (Carthamus tinctorius L.)

Six polyacetylenes have been isolated from immature seeds of safflower (Carthamus tinctorius L.) by thin-layer chromatography. They were identified as 1,11-tridecadiene-3,5,7,9-tetrayne, 1,3,11-tridecatriene-5,7,9-triyne, 1,3,5,11-tridecatetraene-7,9-diyne, 1-tridecene-3,5,7,9,11-pentayne, 1,3-tridecadiene-5,7,9,11-tetrayne and 1,3,5-tridecatriene-7,9,11-triyne from the results of their spectroscopic and chemical analyses. Three of these polyacetylenes had not been isolated from Carthamus tinctorius L.

Changes in the polyacetylene content during maturation were followed by the measurement of ultraviolet absorbance. While 1,3,11-tridecatriene-5,7,9-triyne and 1,3,5,11-tridecatetraene-7,9-diyne had already occurred abundantly at the day of flowering, the amounts of the other polyacetylenes reached the maximum values at the fourth to sixth days after flowering. Although the total amounts of the six polyacetylenes were about 0.8 mmole/g lipid at the fourth day after flowering, no polyacetylene was detected in the mature seeds.     

An analysis of association of components of yield and oil in safflower (Carthamus tinctorius L.)

Summary Inter-relationships of various component characters with yield and oil content were analysed using 215 entries of safflower from India and U.S.A. Correlation of capsule number per plant and capsule weight with yield per plant was pronounced and they showed large direct effects on yield. All other components influenced seed yield mainly through these two components. Seed size had little effect on yield while seed number exerted a positive influence. The proportion of hull expressed as per cent of the whole seed revealed a highly significant and inverse relationship with oil content and was mainly responsible for the observed variability in oil content in the material. Although negative association was indicated between seed size and oil content, it was observed to be due to the indirect effect of hull content and not due to direct effect of seed size. Interestingly, yield per plant and its major components, number of capsules and capsule weight, revealed a negligible relationship with oil content. Both direct as well as indirect effects of hull percent and yield per plant were responsible for the favourable effect of seed number on oil content. The correlation of plant height, days to first flowering and total crop growth period with yield and oil content was either negligible or low, offering scope for developing early maturing and dwarf varieties with high yield and oil content. Spine index showed a non-significant association with yield and oil content. Capsule number, capsule weight and hull per cent were observed to be the most important components in breeding for higher yield and oil content.     

Efficiency of early generation selections for yield and related characters in safflower (Carthamus tinctorius L.)

The efficiency of early generation selection for yield and related characters in safflower (Carthamus tinctorius L.) was studied in the F₂, F₃ and F₄ generations. Twenty-five F₂ progenies derived from various crosses were studied. In the F₂ generation, number of capitula per plant (CNSP), number of seeds per capitulum (SPSP), test weight (SWSP), and seed yield (SYSP) were the criteria used for single plant selection. The analysis of variance showed significant differences for all of the characters in the F₂, F₃, and F₄ generations. The analysis of variance in each of the selection classes showed highly significant genotypic differences. A large number of selections in the CNSP and SYSP classes showed greater yield than the check variety. In each class the mean for that particular character showed a positive shift. The observed F₃ and F₄ means for seed yield per plant was higher in SYSP, indicating the effectiveness of single plant selection for yield. Correlated response showed that selection for number of capitula per plant was effective for improvement of yield.     

Genetic assessment of safflower (Carthamus tinctorius L.) collection with microsatellite markers acquired via pyrosequencing method

A genetic evaluation of safflower germplasm collections derived from different geographical regions and countries will provide useful information for sustainable conservation and the utilization of genetic diversity. However, the molecular marker information is limited for evaluation of genetic diversity of safflower germplasm. In this study, we acquired 509 putative genomic SSR markers for sufficient genome coverage using next‐generation sequencing methods and characterized thirty polymorphic SSRs in safflower collection composed of 100 diverse accessions. The average allele number and expected heterozygosity were 2.8 and 0.386, respectively. Analysis of population structure and phylogeny based on thirty SSR profiles revealed genetic admixture between geographical regions contrary to genetic clustering. However, the accessions from Korea were genetically conserved in distinctive groups in contrast to other safflower gene pool. In conclusion, these new genomic SSRs will facilitate valuable studies to clarify genetic relationships as well as conduct population structure analyses, genetic map construction and association analysis for safflower.     

Estimates of broad-sense heritability for seed yield and yield components of safflower (Carthamus tinctorius L.)

This study was carried out to estimate the broad-sense heritability for seed yield and some yield components of safflower (Carthamus tinctorius L.) cultivars. The experimental design was a randomized complete block design with three replications in the 2004 growing season in the Middle Black Sea Region conditions of Turkey. Three safflower cultivars (5-154, Din?er and Yenice) were grown at five locations (Bafra, Ladik, Suluova, Gümü?hacik?y and Osmancik). The heritability for seed yield, plant height, first branch height, number of branch, head diameter, number of seed per head, 1000-seed weight and oil content were estimates as 35%, 93%, 99%, 45%, 21%, 69%, 81% and 59%, respectively. It was found that first branch height was the least affected trait over environments and followed plant height, thousand seed weight and number of seed per head. On the other hand, head diameter, seed yield, number of branch and oil content were the most affected traits versus environmental conditions. The first branch height, plant height and 1000-seed weight could be used to succeed in selection in early generation.     

Development of EST-SSR markers and their utility in revealing cryptic diversity in safflower (Carthamus tinctorius L.)

With the aim of developing additional genomic resources in safflower, a set of 41,011 ESTs of safflower were mined for the presence of SSRs. 18,773 SSR containing ESTs (SSR-ESTs) were identified and were analyzed to remove redundant sequences leading to identification of 8,810 non-redundant SSR-ESTs (categorized into 6104 singletons and 2,706 contigs) having 13,085 non-redundant SSRs. The average number of non-redundant SSRs per EST was 0.32 and they predominantly consisted of dinucleotide (57.7 %), and trinucleotide (37.7 %) repeat motifs. 500 primer pairs were designed for the non-redundant EST-SSRs of which, 151 were tested. 60 markers which gave robust amplicons, were validated in a set of 19 Carthamus lines. A subset of EST-SSR markers, having average polymorphism information content (PIC) ≥0.4 could precisely elucidate the pedigree relatedness among these lines. Further, these markers exhibited high cross-species transferability to five other wild species of Carthamus. The markers reported here would be a valuable addition to existing safflower marker resources aiding in hastening its improvement.     

Generation and analysis of drought stressed subtracted expressed sequence tags from safflower (Carthamus tinctorius L.)

Drought is the most crucial environmental factor that limits productivity of many crop plants. Exploring novel genes and gene combinations is of primary importance in plant drought tolerance research. Stress tolerant genotypes/species are known to express novel stress responsive genes with unique functional significance. Hence, identification and characterization of stress responsive genes from these tolerant species might be a reliable option to engineer the drought tolerance. Safflower has been found to be a relatively drought tolerant crop and thus, it has been the choice of study to characterize the genes expressed under drought stress. In the present study, we have evaluated differential drought tolerance of two cultivars of safflower namely, A1 and Nira using selective physiological marker traits and we have identified cultivar A1 as relatively drought tolerant. To identify the drought responsive genes, we have constructed a stress subtracted cDNA library from cultivar A1 following subtractive hybridization. Analysis of?~1,300 cDNA clones resulted in the identification of 667 unique drought responsive ESTs. Protein homology search revealed that 521 (78?%) out of 667 ESTs showed significant similarity to known sequences in the database and majority of them previously identified as drought stress-related genes and were found to be involved in a variety of cellular functions ranging from stress perception to cellular protection. Remaining 146 (22?%) ESTs were not homologous to known sequences in the database and therefore, they were considered to be unique and novel drought responsive genes of safflower. Since safflower is a stress-adapted oil-seed crop this observation has great relevance. In addition, to validate the differential expression of the identified genes, expression profiles of selected clones were analyzed using dot blot (reverse northern), and northern blot analysis. We showed that these clones were differentially expressed under different abiotic stress conditions. The implications of the analyzed genes in abiotic stress tolerance are discussed in our study.     

盐胁迫下AM真菌对红花耐盐性的影响

在盐胁迫下,采用盆栽方法研究AM真菌对红花植株耐盐生理指标的影响,以不接种为对照。结果表明,在0、0．1％和0．2％浓度NaCl胁迫下,AM真菌促进红花幼苗的生长,接种真菌的红花叶片SOD和CAT活性、脯氨酸和可溶性蛋白的含量都高于不接种处理的,叶片细胞质膜透性和MDA含量则低于不接种处理的,结果证明AM真菌可以提高植物的耐盐性。     

Oil content and lipid composition of safflower (Carthamus tinctorius) irrigated with saline water under greenhouse and field conditions

An intensive process of land deterioration of some regions in Uzbekistan including the Aral Sea basin has led to a significant increase in soil salinity levels and consequently to a considerable reduction of total fertile soil area, as these lands are of little use for plant growth. The area is estimated to be more than 1.4 million ha of seabed. As a result, there was an immediate need to cultivate new crops capable of stopping the movement of sands and the enlargement of saline soils. Safflower (Carthamus tinctorius) is considered to be a moderately salt‐tolerant crop and, as such, one of a few crops well suited to the cropping systems of salt‐affected soils. It is used in Uzbekistan as a reserve crop when the culture of the main crop fails. In spite of the great economic importance of this oil, there is almost no available information in the literature on the effect of salinity on oil quality and its chemical ingredients. The purpose of the present study was to determine, in greenhouse and field experiments, how irrigation with saline water would influence content of oil, lipids and other lipophylic components in safflower. We found that irrigation of safflower with moderate concentrations of saline water seems feasible, as far as oil and lipid composition is concerned. Consequently, safflower might be a potential crop for lands of little use for plant growth in Uzbekistan or other similar sites in the world.     

Effect of salt stress on growth, fatty acids and essential oils in safflower (Carthamus tinctorius L.)

This study examined the influence of salt treatment on the growth parameters (fresh and dry weights), the mineral content (K⁺ and Na⁺), total lipid contents, fatty acid composition, yields and chemical composition of the essential oil of safflower (Carthamus tinctorius L.) grown in hydroponics for 2 weeks. Results showed that the application of 50 mM NaCl reduced the fresh weight of aerial parts (shoots and leaves) while it enhanced those of the roots. The reduction of dry weight was found to be more pronounced in the aerial parts. Salt treatment increased markedly the concentrations of Na⁺ in both plant parts while it reduced those of K⁺ which resulted in a sharp reduction of K⁺/Na⁺ ratio. In response to salt treatment, total lipids contents decreased in both plant parts and great qualitative changes in the fatty acids profiles were observed. Whatever the plant parts analysed, a redirection of the lipidic metabolism towards synthesis of unsaturated fatty acids as revealed by the increase of double bond index and linoleic desaturation ratio was pointed out. The increased unsaturation index was found to be more important in roots than in aerial parts. Such treatment also reduced the essential oil yields and induced marked quantitative changes in the chemical composition of the essential oils from both plant parts. Of all the identified components, oxygenated components display a prominent salt-induced synthesis and/or accumulation in both roots and aerial parts.