羟基红花黄色素A对脂多糖作用后血管内皮细胞诱导型一氧化氮合酶表达的影响

旨在探讨羟基红花黄色素A(hydroxysafflor yellow A,HSYA)对脂多糖(lipopolysaccharide,LPS)作用后人脐静脉内皮细胞株HUVECs细胞株诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)表达的影响.培养HUVECs细胞株,用 1 mg/L LPS及不同浓度的HYSA处理细胞24 h,MTT法检测细胞增殖情况,硝基还原酶法检测培养液中一氧化氮(NO)含量,RT-PCR及Western blotting检测iNOS表达.结果表明0.01、0.1 mmol/L HYSA对LPS引起的iNOS升高无明显作用,但1 mmol/L HYSA能明显抑制LPS作用后高度表达的iNOS量.因此,HYSA能下调LPS所致iNOS的异常表达,这可能有助于临床治疗血管炎症疾病.     

不同氮源对小麦幼苗谷氨酰胺合成酶的影响

利用ＤＥＡＥ－纤维素柱层析、酶活性测定、Ｎｏｒｔｈｅｒｎ 分子杂交等技术,研究了小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．）幼苗的根、叶和离体叶在不同氮源培养条件下谷氨酰胺合成酶（ＧＳ）活性和同工酶变化, 以及不同氮源对ＧＳ基因转录－ＧＳ－ｍ ＲＮＡ 的影响． 同时与硝酸还原酶（ＮＲ）活性进行比较, 结果表明∶当以ＮＨ＋４ 作唯一氮源时,小麦幼苗根谷氨酰胺合成酶（ＧＳｒ）和叶细胞质谷氨酰胺合成酶（ＧＳ１）活性要比以ＮＯ－３ 作唯一氮源的高．当以ＮＯ－３ 为唯一氮源时, ＮＯ－３ 则促进完整叶片和离体叶片叶绿体谷氨酰胺合成酶（ＧＳ２）活性． 从转录水平上看,ＮＨ＋４ 促进根ＧＳ－ｍ ＲＮＡ 的合成,而ＮＯ－３ 促进叶ＧＳ－ｍ ＲＮＡ 的合成     

不同氮源对球形棕囊藻生长的影响

采用实验室培养的方法比较了6种不同氮源-硝态氮、尿素、甘氨酸、精氨酸、谷氨酸、腺嘌呤对典型赤潮藻球形棕囊藻(Phaeocystis globosa)生长的影响。结果表明,6种氮源均能不同程度地促进球形棕囊藻的生长,但比生长速率和光合作用效率具有显著差异性。将球形棕囊藻在不同浓度氮源下的最大比生长速率分别拟合Monod方程,得出球形棕囊藻在硝态氮、尿素、甘氨酸、精氨酸、谷氨酸和腺嘌呤等6种氮源下的最大比生长率分别为1.05,1.17,0.82,0.87,1.09,0.90d^-1,相应的半饱和常数分别为9.132,23.758,85.519,7.104,23.94,10.959μmol/L。其中,高氮浓度(8820μmol/L)下腺嘌呤对球型棕囊藻的生长具有显著抑制作用。相比较而言,球形棕囊藻对甘氨酸的亲和力最高。当硝态氮、尿素、甘氨酸、精氨酸、谷氨酸和腺嘌呤的浓度分别为8820,882,882,8820,882,0.441μmol/L时,球形棕囊藻的最大光合效率(Fv/Fm)分别为0.619,0.620,0.579,0.595,0.648,0.667。由此可见,氮源对球形棕囊藻的生长和光合作用具有显著影响;球形棕囊藻能够利用多种无机和有机氮源,与其它仅能利用无机氮源的浮游植物相比,更具有竞争优势。     

alpha- 乳香酸联用羟基红花黄色素A抗胃溃疡效果研究

目的：研究乳香红花提取物中主要药效物质alpha-乳香酸（alpha-BA）和羟基红花黄色素A（HSYA）联合应用的胃保护效果。方法：36 只SD 大鼠随机分为6 组：正常组（生理盐水5 mL/kg）,模型组（生理盐水5 mL/kg）,西咪替丁组（100 mg/kg）,HSYA 组（100mg/kg）,alpha-BA 组（200 mg/kg）,联合用药组（HSYA 50 mg/kg+alpha-BA 100 mg/kg）。预给药1 小时后,用乙醇诱导SD 大鼠胃溃疡造模,评价溃疡指数、胃内容物酸度、胃壁黏液量,用ELISA法检测血清炎性因子TNF-alpha、IL-1beta、IL-8。结果：①与造模组相比,alpha-BA组和联合用药组显著降低乙醇诱导胃粘膜损伤（P＜0.001）且联合用药组较西咪替丁组相比差异无统计学意义,但HSYA组差异无统计学意义;与造模组相比,alpha-BA组和联合用药组显著升高胃内容物pH,增加胃黏液含量（P＜0.001）且联合用药组与西咪替丁组相比差异无统计学意义,但HSYA 组仅胃黏液含量有一定升高（P＜0.05）,胃内容物pH 没有明显改变。②与造模组相比,HSYA 组、alpha-BA组、联合用药组均显著抑制血清TNF-alpha、IL-1beta、IL-8 的升高（P＜0.001）且联合用药组与西咪替丁组相比差异无统计学意义。③H&E 染色观察可见HSYA组依然有明显出血性损伤;西咪替丁组、琢-BA组和联合用药组则明显保护胃黏膜、抑制损伤和白细胞浸润。结论：alpha-BA有较好的胃保护效果,HSYA胃保护效果显著低于alpha-BA,但联合应用HSYA、alpha-BA能够抑制胃黏膜出血、白细胞浸润、抑制胃内容物酸度,增加胃黏液含量,并且能够调节血清炎性因子从而发挥较单用药用量更少且效果更好的胃保护作用。     

稀土对红花幼苗生长及部分酶同工酶的影响

用不同浓度的混合稀土Re(NO3)2处理红花幼苗,测定稀土元素（RE）对红花幼苗生长及其部分酶同工酶的影响。结果表明：RE对红花幼苗生长的影响是适宜浓度起促进,氏于适宜浓度作用不明显,高于适宜浓度有抑制作用;RE能提高幼苗地上部分淀粉酶,酯酶和过氧化的酶的活性,特别是淀粉酶的活力增强显著,但对过氧化物酶的影响不明显。     

不同氮源对蛋白核小球藻生长、色素和中性脂肪积累的影响

单细胞微藻在生长发育过程中,所积累的中性脂肪具有潜在的生物燃料价值。不同氮源对藻类的生长具有显著影响。研究了氨态氮、尿素氮和硝酸态氮对蛋白核小球藻生长、色素和中性脂肪积累的影响。结果显示,不同氮源对培养液pH有显著影响,以氨态氮为氮源导致培养液pH降低,而硝酸态氮导致培养液pH升高,培养液pH的波动可被添加的Hepes所稳定,并促进以氨态氮为氮源的蛋白核小球藻的生长。尿素氮对蛋白核小球藻生长、色素积累的效果优于氨态氮和硝酸态氮,硝酸态氮在中性脂肪积累上优于尿素氮和氨态氮,添加Hepes对氮饥饿后蛋白核小球藻的中性脂肪积累无显著影响。     

超声辅助提取红花中主要活性成分的工艺优化

采用HPLC-UV测定红花提取物中黄酮类成分含量;确定评价指标;通过单因素试验设计;筛选红花活性成分提取方法;利用Box-Behnken试验设计原理;以花瓣中主要黄酮类成分为响应值;以单因素结果所选因素为自变量;建立响应面分析试验模型;优化提取工艺。红花花瓣中主要黄酮类成分为羟基红花黄色素A和山奈酚-3-O-芸香糖苷;占比85%以上;红花活性成分提取方法应采用超声提取法;响应面优化的最佳提取条件为:料液比0.1∶25 g·mL^-1、超声温度70℃、超声时间45 min、溶剂为57%甲醇、超声功率177 W;在该最优条件下红花黄色素A提取率为1.74%;山奈酚-O-β-芸香糖苷提取率为1.53%。该方法羟基红花黄色素A提取率比药典中提取方法显著提高;其余黄酮类成分提取率也呈极显著增加。     

水位变化对南荻幼苗生长的影响

以洞庭湖优势植物南荻(Miscanthus lutarioriparius)为研究对象, 探讨了三种水位[ 0 cm(对照组)、20 cm、40 cm ]下, 幼苗经40天淹水胁迫和40天退水恢复后(水位恢复到0 cm)的形态、生物量、可溶性糖及淀粉含量变化。结果表明, 水位对南荻幼苗生长以及其生理特征有显著的影响。经淹水胁迫后, 随着水位的升高, 南荻采取静止生存策略, 并呈现出株高、根长减小以及生物量和可溶性糖含量降低的趋势。经退水恢复后, 仅20 cm低水位下根状茎生物量恢复至胁迫后的80.3%, 其余水位下幼苗生长生理参数与胁迫后相比均有所升高。由此表明, 40天的20 cm低水位淹水胁迫对南荻生长发育影响较小, 而40 cm高水位胁迫会导致其生长受到严重抑制。本研究结果可揭示挺水植物南荻对洪水的适应性, 为南荻恢复提供参考依据。     

外加氮源对葛仙米生长及光合生理的影响

分别测量培养在不同氮浓度培养基中的葛仙米叶绿素a含量和3种叶绿素荧光参数(Fv/Fm、(φ)PS Ⅱ、ETR),研究外加氮源对葛仙米生长和光合生理的影响.结果表明:外加氮源抑制葛仙米的生长,并且随着浓度升高,抑制效果更加明显.外加氮源浓度小于等于1.5 g/L时,葛仙米光合电子传递速率(ETR)、PS Ⅱ最大量子产量(Fv/Fm)和PS Ⅱ光化学反应量子效率((φ)PS Ⅱ)值均有先下降后上升趋势;而当外加氮源浓度大于1.5 g/L时,上述3种荧光参数值先维持不变后有下降趋势.     

几种不同碳、氮源对隐甲藻生长及二十二碳六烯酸产量的影响

目的：找出有利于隐甲藻生长和二十二碳六烯酸（DHA）积累的碳、氮源。方法：利用不同碳、氮源培养隐甲藻,收集藻体后提取脂肪酸并甲酯化,然后利用毛细管气相色谱法进行分析。结果：最适的单一碳、氮源分别为葡萄糖、酵母粉,在此培养条件下隐甲藻培养72h后的生物量（干重）和DHA产量分别为3．90和0．642g／L。结论：葡萄糖、酵母粉分别作为碳、氮源时更有利于隐甲藻的生长和DHA的积累。     

Identification of AFLP fragments linked to hydroxysafflor yellow A in Flos Carthami and conversion to a SCAR marker for rapid selection

Hydroxysafflor yellow A (HSYA), an important active compound in treating focal cardiac and cerebral ischemia, is uniquely present in flower petals of Carthamus tinctorius. In this study, inheritance and molecular marker analyses for HSYA trait in safflower were carried out. HSYA contents in parents, cross hybridized F₁ and F₂ individuals were analyzed by high performance liquid chromatography. Results revealed that the presence/absence of HSYA was controlled by one major nuclear gene termed HSya. A total of 48 AFLP primer combinations were screened, and bulked segregant analysis was performed by preparing two pools of 10 present-HSYA and ten absent-HSYA plants selected from the 498 individuals of the F₂ segregating population. Four AFLP markers, AFLP-5, AFLP-7, AFLP-15 and AFLP-16, were identified to be closely associated with HSya. Of those, AFLP-16 was the closest to HSya, estimated at about 9.4 cM in genetic distance. The dominant AFLP-16 marker was converted into a simple sequence characterized amplified region marker based on the sequence information of the cloned flanking regions of the AFLP fragment and was designated as SCM16. Our result has direct application for marker-assisted selection of quality breeding in safflower.     

Expression of CT-wpr, screened by cDNA-AFLP approach, associated with hydroxysafflor yellow A in Carthamus tinctorius L.

Hydroxysafflor yellow A (HSYA) is the main active compound in petals of Carthamus tinctorius L. In this study, we used cDNA-amplified fragment length polymorphism (cDNA-AFLP), coupled with bulked segregant analysis (BSA), to screen differentially expressed genes in two cDNA pools, and obtained 2560 reproducible bands ranging from 1500 bp to 100 bp with 64 AFLP primer combinations. Two TDFs (TDF-4 and TDF-11) were identified with low recombination and successfully sequenced. The genetic distance of the linkage map from the HSya locus was 9.9 cM and 2.9 cM, respectively. The full-length cDNA sequence of TDF-11 gene, designated as CT-wpr (GenBank Accession No. GU227360) was cloned. Nucleotide sequence alignment revealed that CT-wpr shared 66% identity with Populus trichocarpa. Semi-quantitative RT-PCR analysis revealed that the mRNA of the two TDFs only expressed in the HSYA-absent line. Possible functions of CT-wpr are also discussed.     

Direct somatic embryogenesis and plantlet regeneration from cotyledonary leaves of safflower

Somatic embryos were induced directly on adaxial surface of cotyledonary leaves within 8–10 days of culture on Murashige and Skoog medium containing 5.37 to 10.74 M 1 — napthaleneacetic acid and 2.22 M benzyl adenine. Germinated embryos with shoot axes developed into complete plants after transfer onto half stength Murashige and Skoog medium containing 1.07 M 1 — napthaleneacetic acid. Histological studies suggested direct origin of somatic embryos with broad-base attachment.Abbreviations BA benzyl adenine - MS Murashige and Skoog - NAA 1-napthaleneacetic acid - RH relative humidity     

Abscisic acid, temperature and salinity interactions on growth and some mineral elements in Carthamus plants

Growth and contents of sodium (Na), potassium (K), calcium (Ca), magnesium (Mg), chloride (Cl), phosphorus (P) and sulphur (S) in shoot and root tissues of Carthamus tinctorius plants were measured at combinations of four nutrient solution osmotic potentials (_s=0, -0.3, -0.6 and -0.9 MPa) induced by NaCl and CaCl treatments, three constant temperatures (T) ranging from 15 to 35°C and four abscisic acid (ABA) concentrations (0,10,50 and 100 mg L^–1). Unstressed and stressed plants grown in optimal temperature conditions (25°C) maintained higher growth rates (dry mass production) than plants grown under low and high temperatures (15 and 35°C respectively). Shoot and root growth (dry mass production) were largely inhibited by salinity but the magnitude of growth inhibition was temperature dependent. Safflower plants respond to salinity stress by increases in Ca, Cl and to a lesser extent Na in their shoots and roots and by a decrease in the ratio of fresh to dry weight. The ratio of K/Na was decreased progressively on salinization. With stressed plants, ABA application reduced the toxicity of salt treatment, improved K uptake under salinity, effectively increased K/Na ratio and helped the plants to avoid Na toxicity and sometimes enhanced growth. The effect of ABA on the growth was more pronounced at optimum temperature (25°C). The association between the internal mineral element concentrations was largely affected by ABA application and temperature change but a wide fluctuation in response was noticed. The effects of single factors (_s, T and ABA) on the growth and mineral contents were statistically significant. Also, bifactorial (_s× T, _s × ABA and T × ABA) and three factorial (_s × T × ABA) interactions significantly affected the parameters. Further statistical treatment of the data (coefficient of determination ²) led to four important findings: (1) Salinity (_s) was dominant in affecting Ca and Cl contents in both shoot and root as well as root Na content. (2) Temperature (T) had a dominant effect on growth, shoot K, Mg, P, S and root P, and S contents (3) The share of _s × T × ABA interaction was dominant for root Na and Mg contents. (4) The single factors and their interactions had a dual role in their subsidiary effects.Abbreviations ABA abscisic acid - _s osmotic potential - ² coefficient of determination - F.wt fresh weight - d.m. dry matter - T temperature - MPa mega pascal - SAR sodium adsorption ratio - P phosphorus - S sulphur     

Behavior of cell aggregate of Carthamus tinctorius L. cultured cells and correlation with red pigment formation

The formation processes of Carthamus tinctorius cell aggregates in a growth medium and the correlation of red pigment formation with cell aggregate sizes were investigated. About 80% of cell aggregates in the growth medium were > 1.00 mm in size. The growth rate of large cell aggregates was more rapid than that of small cell aggregates. Most cell aggregates > 0.50 mm in size became larger or smaller than their original sizes during the culture. A high level of red pigment formation was observed when cell aggregates obtained by the preculture using cell aggregates < 1.00 mm in size were cultured in the production medium.     

Thidiazuron stimulates adventitious shoot regeneration in different safflower explants

Adventitious shoot regeneration from root, hypocotyl, cotyledon and primary leaf explants of safflower (Carthamus tinctorius L.) was studied. Shoot regeneration was promoted by benzyladenine (BA) + naphthaleneacetic acid (NAA), BA + indole-3-butyric acid (IBA), kinetin + NAA and thidiazuron (TDZ) + NAA incorporated in Murashige and Skoog (MS) basal medium. High frequency of shoot regeneration and high number of shoots per regenerating explant were obtained on a wide range of TDZ + NAA combinations. Proliferated shoots were elongated in MS + 0.5 mg dm⁻³ kinetin and well-developed shoots were rooted in half strength MS + 0.5 mg dm⁻³ NAA. Rooted shoots were successfully acclimatized and established in soil.     

Gibberellin physiology of safflower: endogenous gibberellins and response to gibberellic acid

Endogenous gibberellins (GAs) were extracted from safflower (Carthamus tinctorius L.) stems and detected by capillary gas chromatography-mass spectrometry from which GA₁, GA₃, GA₁₉,, GA₂₀, GA₂₉, and probably, GA₄₄ were detected. The detection of these GAs suggests that the early 13-OH biosynthetic pathway is prevalent in safflower shoots. Deuterated GAs were used as internal standards and GA concentrations were determined in stems harvested at weekly intervals. GA₁ and GA₁₉ levels per stem increased but concentrations per gram dry weight decreased over time. GA₂₀ was only detected in young stem tissue.Gibberellic acid (GA₃) was also applied in field trials and both GA₃ and the GA biosynthetic inhibitor, paclobutrazol, were applied in growth chamber tests. GA₃ increased epidermal cell size, internode length, and increased internode cell number causing stem elongation. Conversely, paclobutrazol reduced stem height, internode and cell size, cell number and overall shoot weight. In field tests, GA₃ increased total stem weight, but decreased leaf weight, flower bud number and seed yield. Thus, GA₃ promoted vegetative growth at the expense of reproductive commitment. These studies collectively indicate a promotory role of GAs in the control of shoot growth in safflower, and are generally consistent with gibberellin studies of related crop plants. Author for correspondence