In vitro induction of androgenic haploids in Safflower (Carthamus tinctorius L.)

The influence of culture medium on induction of androgenic calli was examined with five different basal media. MS medium was the most responsive in inducing callus. Differences in induction of calli among ten genotypes revealed that the most responsive genotype was a local cultivar, Mangira, with 48.6% anthers initiating callus formation. The influence of temperature pre-treatment (5°±1°C) for varying periods (0 to 15 days) on immature capitula prior to inoculation of anthers on the medium revealed that the percentage of anthers inducing callus increased till 3–5 days of pre-treatment. The effect of physiological conditions of anther donor plants grown in the field and in green houses on induction and re-differentiation have shown that the field grown anther donor plants exhibited optimum response. Shoot regeneration was observed on MS supplemented with BAP (2.0 mg/l) and NAA (0.5 mg/l) and rhizogenesis on MS (half-strength) medium, supplemented with NAA (0.1 mg/l) and 1% sucrose. Cytological studies of anther derived plants showed two ploidy levels, where the haploids were predominant (64%).     

Antioxidant and Neuroprotective Activities of Mogami-benibana (Safflower,Carthamus tinctorius Linne)

Free radical scavenging activity of the extracts of petals (bud, early stage, full blooming and ending stage), leaf, stem, root and seeds of Mogami-benibana (safflower, Carthamus tinctorius Linne), the contents of the major active components of carthamin and polyphenols, and neuroprotective effect of the petal extracts and carthamin in the brain of mice and rats were examined. Water extracts of Mogami-benibana petals scavenged superoxide, hydroxyl and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and singlet oxygen. The scavenging activities of the extract of safflower petals with various colors showed the order of orange, yellow and white from high to low. This order is consistent with the contents of carthamin, which is a pigment of orange color and is found highest in orange petals and lowest in white petals. There was also a relationship between DPPH radical scavenging activity and carthamin content in the petal extracts of safflower. The neuroprotective effects were examined in cellular and animal models. Mogami-benibana petal extract inhibited glutamate-induced C6 glia cell death, significantly decreased the formation of malondialdehyde in mouse cerebrum, and inhibited the increase in thiobarbituric acid reactive substances and 8-hydroxy-2′-deoxyguanosine (8-OHdG) in the cerebral cortex of rats subjected to an injection of FeCl₃ solution into the sensory motor cortex. Carthamin showed similar effects in inhibiting 8-OHdG by the petal extract in rats. These results suggest that the petal extract of Mogami-benibana has free radical scavenging activity and neuroprotective effect and carthamin is one of the major active components. Special Issue Article in Honor of Dr. Akitane Mori.     

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.     

Serotobenine,a Novel Phenolic Amide from Safflower Seeds (Carthamus tinctorius L.)

The structure of a new indole derivative, serotobenine (1), from sfflower meal (Carthamus tinctorius> L.) is proposed based on ¹H- and ¹³C-NMR spectral data and X-ray crystallography. The known compounds N-feruloyltryptamine (2) and N-(p-coumaroyl)tryptamine (3) were also isolated and identified.     

Conjugated Serotonins and Phenolic Constituents in Safflower Seed (Carthamus tinctorius L.)

New three conjugated serotonins were isolated from safflower meal (carthamus tinctorius L.). On the basis of spectral properties and chemical evidence, their structures were determined as N-feruloylserotonin (4), N-(p-coumaroyl)serotonin (5) and N-(p-coumaroyl)serotonin mono-β-d-glucopyranoside (6), The known compounds, 2-hydroxyarctiin (2), matairesinol mono-β-d-glucoside (1) and acacetin (3) were also isolated and identified.     

甜椒的离体培养及建立再生体系研究

以荷兰进口甜椒的子叶和下胚轴为外植体,接种到附加不同植物生长调节剂的培养基上,筛选出MS+6-BA 4.0 mg/L+NAA 0.1 mg/L为子叶和下胚轴最佳不定芽分化培养基;MS+IBA 0.2 mg/L和MS+IBA 0.5 mg/L为最佳不定根分化培养基。     

云南红花种质资源主要农艺性状的遗传多样性分析

为加强红花种质资源的研究利用,对筛选出的66份云南红花优异种质资源16个形态性状进行聚类分析与主成分分析。结果表明:云南红花种质资源具有丰富的遗传多样性,多样性指数最高的是果球着粒数,其次是株高、最末分枝高度和千粒重;性状变异系数最大的是分枝总数,其次分别是单株有效果球数和第一分枝高度,最小的为顶果球直径;基于各种质间形态性状的遗传差异,把66份红花种质聚类并划分为6大类群。第Ⅰ类群可作为有增产潜力的亲本材料,第Ⅲ类群可作为高产量目标选育的亲本,第Ⅳ类群可作为大粒型选育亲本,第Ⅴ类群可作为高含油量选育目标亲本,第Ⅵ类群既是大粒型又是高含油量双重选育目标亲本。11个数量性状的主成分分析结果表明,前4个主成分累计贡献率达82.59%,第一主成分反映植株高度,第二主成分反映产量构成因子,第三、第四主成分分别反映千粒重和果球着粒数。研究结果表明云南红花地方种质资源的变异较大,遗传较丰富。     

Embryo Transformation, A Practical Approach for Realizing Transgenic Plants of Safflower (Carthamus tinctorius L.)

A gene transfer system that ensured recovery of whole planttransformants was developed for safflower (Carthamus tinctoriusL.). Embryo axes of germinating seeds with one of the cotyledonsremoved were pricked with a sterile sewing needle at the cotyledonarynode and infected by gentle agitation for 10 min in a suspensionofAgrobacterium tumefaciens . Following a 24 h co-cultivationand decontamination with cefotaxime for 1 h, they were placedon soilrite moistened with water to allow germination to progress.Later, the seedlings were transferred to soil in pots wherethey grew into normal healthy plants in the greenhouse. Thehistochemical assay of an uid A gene that expresses only inplant tissues and PCR amplification of uid A and npt II markergenes were used for early determination of putative transformants,whereas Southern analysis of T₀and T₁plant DNA was used to confirmintegration of the transgenes. The combined results indicatedthat the frequency of transformation was 5.3% in safflower ‘A-1’and 1.3% in ‘A-300’. Four T₀plants of ‘A-1’yielded transformed T₁progeny. The strategy, in principle, shouldbe applicable to all cultivars and genotypes of safflower whichare susceptible to Agrobacterium tumefaciens infection. Thusfar, this is the only procedure available for safflower thatcould successfully be used to generate whole plant transformants.Copyright 2000 Annals of Botany Company Safflower (Carthamus tinctorius L.), transformation, non-tissue culture method, embryo axes,Agrobacterium , transgene expression     

In Vitro Culture and Plant Regeneration in Vicia faba subsp. Equina (var. Spring Blaze)

Explants of stem, leaves, roots, and cotyledons from etiolatedaxenically grown Vicia faba seedlings were cultured on a rangeof media. Shoot organogenesis was only obtained with nodal stemand cotyledonary node explants when cultured on MS medium with3% sucrose, 2.0 mg 1^–1 BAP and 02 mg 1^–1 NAA. Callusproliferation accompanied shoot organogenesis from nodal stemexplants. Successive subculture of nodal stem callus resultedin proliferation of regenerative callus which contained severalshoot bud initials. The capacity for shoot regeneration fromthis callus was maintained for 9 months. Histological studiesreveal de novo formation of meristematic centres in callus andtheir further development into bud primordia. High frequencyrooting of these adventitious shoots was obtained on half-strengthMS medium with 1.5% sucrose, 0.1 mg 1^–1 NAA and 0.5 mg1^–1 kinetin. Key words: Vicia faba, adventitious shoots, axillary shoots, de novomeristem formation, organogenesis, tissue culture     

世界红花种质的籽油脂肪酸组分评价

对引自 48个国家和地区在北京栽培的 2 0 48份红花 (CarthamustinctoriusL .)种质资源的籽油脂肪酸分析表明 ,棕榈酸、硬脂酸、油酸和亚油酸的平均含量分别为 7.30 %、1.2 8%、15 .76 %和 75 .33% ,其含量范围分别为 0 .99%～ 2 9.0 3%、0 .0 1%～ 5 .71%、5 .0 0 %～ 81.84%和 11.13%～ 88.30 %。来自不同地区的红花种质 ,各种脂肪酸的含量有较大的差异。来源于孟加拉国的红花 ,亚油酸平均含量为 5 0 .6 8% ,来源于奥地利的红花 ,亚油酸平均含量高达79.0 4%。通过评价 ,分别筛选出 10个高亚油酸和 10个高油酸的品种 ,高油酸的品种中有 3个来自孟加拉国 ,而高亚油酸的品种大多来自中国     

In Vitro Regeneration of Mat Sedge (Cyperus pangorei Rottb)

An in vitro protocol was developed for regeneration of Cyperus pangorei that may supplement enough raw materials for the mat weaving community. Callus was initiated from inflorescence explants on Murashige and Skoog’s (MS) medium supplemented with 5 and 10 μM each of 2, 4-D, 2, 4, 5-T and CPA. Development of numerous de novo spikelets from immature inflorescence explants grown in (10 μM) 2, 4, 5-T was observed. MS with 5 μM Kn and 100 ml l^?1 Coconut milk (CM) promoted shoot regeneration from calli. Calli from 2,4-D and CPA medium sub-cultured on medium containing 5 μM BAP, 5 μM Kn, 1 μM IAA and 100 ml l^?1 CM produced extensive and rapid rhizogenesis with wiry and scaly roots. Micropropagation using rhizome buds on MS medium with BAP, Kn and Zeatin at 10 μM concentrations resulted in shoot release and multiplication by breaking the bud dormancy. An average of 10 shoots per explant was produced in 10 μM BAP, whereas (10 μM) Kn and (10 μM) Zeatin induced only single shoot formation. The shoots were transferred to rooting media comprising 10 μM IAA with 1 μM BAP or Kn and then acclimatized. The results accomplished were found to be useful in developing a complete in vitro regeneration protocol towards the mass production of Cyperus species, which may provide a basis for further genetic improvements that may prove its use as an alternative natural fibre resource in commercial applications.     

辣椒离体培养及再生体系的研究

选用9个辣椒(Capsicum annuumL.)品种(系),研究了不同激素组合、基因型、外植体类型、苗龄和Ag-NO3等因素对外植体不定芽分化和伸长的影响.结果表明,在6-BA/IAA为10∶1配比下,有利于辣椒外植体的分化再生,而6-BA/IAA为3∶1配比下适合于再生芽的伸长;不同品种辣椒的再生能力差别较大,分化率在13.3%~90.0%之间;辣椒子叶再生能力比下胚轴强,是较好的外植体材料;12~16 d苗龄的外植体分化频率较高;添加4mg?L-1AgNO3可使芽分化率平均提高16.9%.通过比较,筛选出了适合于辣椒芽分化的培养基为MB5(MS无机盐 B5有机成分) 5 mg?L-16-BA 0.5 mg?L-1IAA 4 mg?L-1AgNO3,芽伸长培养基为MB5 3 mg?L-16-BA 1 mg?L-1IAA 2 mg?L-1GA3 4 mg?L-1AgNO3,生根培养基为1/2 MS 0.2 mg?L-1IAA 0.1 mg?L-1NAA.     

Effect of acidicpH on the protein carmin from safflower seed (Carthamus tinctorius)

The effect of a decrease inpH on the structural integrity of carmin has been monitored by a variety of biophysical techniques. The protein undergoes initial dissociation up topH 3.5–4.0 without any significant denaturation. Below thispH the process of dissociation and denaturation appears to be simultaneous. Further, in thepH range of 2.5–1.6 the protein reassociates to probably a different polymer resulting from possibly, an entropically driven hydrophobic interaction. The process of dissociation appears to be reversible to a large extent. The process of denaturation appears to be governed by the kinetic path that the denatured protein molecule follows either by a sudden decrease inpH or through a gradual decrease inpH. These results are interpreted while keeping in view the oligomeric and globular structure of carmin at neutralpH. The results would help in understanding of structure-function relationship of the protein and its role in hydrogen ion bindingin vivo.     

In Vitro Regeneration of Stevia rebaudiana (Bert) from the Nodal Explant

Procedure for micropropagation of Stevia rebaudiana Bertoni, containing stevioside, a natural noncaloric sweetner, has been developed using nodal segments as explant. Higher proliferation of shoots and multiplication was obtained on Murashige and Skoog basal medium (MS) supplemented with 1.0 mg l-1 indoleacetic acid (IAA) plus 10.0 mg l^-1 kinetin and 30.0 mg l-1 adenine sulphate. Sprouting of 90% of the axillary buds was observed within 4 weeks of inoculation, producing >10.0 shoots per explant within 12 weeks. Profuse roots were induced from 90% of the regenerated shoots within 4 weeks of inoculation on half strength MS solid medium supplemented with 1.0 mg l^-1 IAA. High survival rate, > 60%, was obtained when the plantlets were transferred to field conditions. The survival rate of taller plants was always higher. The in vitro regenerated plants were morphologically indistinguishable from the donor plants and leaves were of intense sweet taste upon chewing. The heterogenic nature of S. rebaudiana necessitates establishment of protocol for every genotype independently.     

In Vitro Regeneration of European Linden

The regeneration of European linden (Tilia × europaea L.) in vitro was successful. When using axillary buds as primary explants it was possible to induce a new shoot growth. The highest number of shoots per explant (2.13 ± 1.09) was recorded at the presence of 0.2 mg dm^–3 6-benzylaminopurine. Up to 50% of elongated shoots rooted in the presence of 2.0 mg dm^–3 -naphthaleneacetic acid.     

Genetic characterization of Iranian safflower (Carthamus tinctorius) using inter simple sequence repeats (ISSR) markers

Safflower (Carthamus tinctorious L.) is valued as a source of high quality vegetable oil. 20 ISSR primers were used to assess the genetic diversity of 18 accessions of safflower collected from different geographical regions of Iran. The ISSR primers combinations revealed 57.6 % polymorphism, among 338 genetic loci amplified from the accessions. The sum of effective number of alleles and observed number of alleles were 29.76 and 36.77, respectively. To understand genetic relationships among these cultivars, Jacquards’ similarity coefficient and UPGMA clustering algorithm were applied to the ISSR marker data set. ISSR markers grouped accessions into two main clusters and four sub clusters. Also, the principal coordinate analysis (PCoA) supported the cluster analysis results. The results showed these genotypes have high genetic diversity, and can be used for alternative safflower breeding program.