大孔树脂分离纯化“黑金刚”紫马铃薯花青苷工艺研究

以紫色马铃薯"黑金刚"花青苷为原料,采用D101、HDP100A、HDP450A、NK-9、AB-8五种大孔吸附树脂对花青苷的吸附与解析特性进行了比较研究,并在此基础上,采用最佳大孔树脂对花青苷纯化过程中的静态、动态吸附和解析附条件进行了优化研究。结果表明AB-8大孔树脂具有较好的吸附和解析能力,是纯化紫色马铃薯花青苷的最佳树脂,较优纯化条件为:上样液花青苷浓度为0.028mg.g-1,上样液pH=2,洗脱液乙醇浓度为50%,洗脱液pH=1,吸附流速为1mL.min-1,洗脱流速为1mL.min-1。经大孔树脂纯化后,色价值比纯化前提高了7.55倍。     

Assessing leaf pigment content and activity with a reflectometer

This study explored reflectance indices sampled with a 'leaf reflectometer' as measures of pigment content for leaves of contrasting light history, developmental stage and functional type (herbaceous annual versus sclerophyllous evergreen). We employed three reflectance indices: a modified normalized difference vegetation index (NDVI), an index of chlorophyll content; the red/green reflectance ratio ( R _RED: R _GREEN), an index of anthocyanin content; and the change in photochemical reflectance index upon dark–light conversions (ΔPRI), an index of xanthophyll cycle pigment activity. In Helianthus annuus (sunflower), xanthophyll cycle pigment amounts were linearly related to growth light environment; leaves in full sun contained approximately twice the amount of xanthophyll cycle pigments as leaves in deep shade, and at midday a larger proportion of these pigments were in the photoprotective, de-epoxidized forms relative to shade leaves. Reflectance indices also revealed contrasting patterns of pigment development in leaves of contrasting structural types (annual versus evergreen). In H. annuus sun leaves, there was a remarkably rapid increase in amounts of both chlorophyll and xanthophyll cycle pigments along a leaf developmental sequence. This pattern contrasted with that of Quercus agrifolia (coast live oak, a sclerophyllous evergreen), which exhibited a gradual development of both chlorophyll and xanthophyll cycle pigments along with a pronounced peak of anthocyanin pigment content in newly expanding leaves. These temporal patterns of pigment development in Q. agrifolia leaves suggest that anthocyanins and xanthophyll cycle pigments serve complementary photoprotective roles during early leaf development. The results illustrate the use of reflectance indices for distinguishing divergent patterns of pigment activity in leaves of contrasting light history and functional type.     

Anthocyanin pigments in Callistephus chinensis

Identification of the anthocyanin pigments in the flowers of six genotypes of Callistephus chinensis has confirmed that a series of multiple alleles, R, r′ and r are responsible for the production of delphinidin, cyanidin, and pelargonidin derivatives respectively. However, mixtures of anthocyanidin types were present in all genotypes. In the presence of gene M, mainly 3,5-diglycosides were found; in recessive genotypes (mm) there were only 3-mono-glucosides. Unstable acylated derivatives of these pigments were also present.     

Safety and whole-body antioxidant potential of a novel anthocyanin-rich formulation of edible berries

Edible berry extracts rich in anthocyanins possess a broad spectrum of therapeutic, pharmacologic and anti-carcinogenic properties. Six berry extracts (wild blueberry, bilberry, cranberry, elderberry, raspberry seeds and strawberry), singly and in combination, were studied in our laboratories for antioxidant efficacy, cytotoxic potential, cellular uptake and anti-angiogenic properties. Combinations of edible berry extracts were evaluated to develop a synergistic formula, OptiBerry, which exhibited high oxygen radical absorbance capacity (ORAC) value, low cytotoxicity and superior anti-angiogenic properties compared to the other combinations tested. The current study sought to determine the broad spectrum safety and antioxidant potential of OptiBerry in vivo. Acute oral LD₅₀ of OptiBerry was greater than 5 g/kg in rats. Acute dermal LD₅₀ of OptiBerry was greater than 2 g/kg. No changes in the body weight or adverse effects were observed following necropsy. Primary skin and eye irritation studies were conducted in New Zealand albino rabbits. OptiBerry was classified as slightly irritating to the skin (primary skin irritation index 0.3) and minimally irritating to the eye (maximum mean total score 6.0). The antioxidant potential of OptiBerry was investigated in rats and mice by assessing GSH redox status in tissues as well as by a unique state-of-the-art electron paramagnetic resonance (EPR) imaging of whole-body redox status. A clinically relevant hyperbaric oxygen (HBO) exposure system (2 atm, 2 h) was employed to study the antioxidant properties of OptiBerry. OptiBerry feeding (8 weeks) significantly prevented HBO-induced GSH oxidation in the lung and liver of vitamin E-deficient Sprague Dawley rats. Furthermore, OptiBerry-fed mice, when exposed to HBO, demonstrated significant protection in whole-body HBO-induced oxidation compared to the unfed controls by EPR imaging. Taken together, these results indicate that OptiBerry is reasonably safe and possess antioxidant properties.     

前体饲喂、诱导子和光照联合使用对葡萄细胞培养合成花青素的影响

苯丙氨酸前体饲喂分别和环糊精、葡聚糖、茉莉酸甲酯、黑曲霉和直喙镰孢菌提取液五种诱导子联合作用,其中以与茉莉酸甲酯的联合作用对葡萄细胞培养生产花青素的影响最大,可使单位鲜细胞花青素含量提高2.7倍,花青素产量提高3.4倍,实验证明两者在培养后第4天加入效果最好。在30μmol/L苯丙氨酸、218μmol/L茉莉酸甲酯和3000~4000lx光照条件下,不同花青素产量的细胞株都能显著提高花青素产量,但低产株VV06比高产株VV05具有更大的产率提高潜力。该条件下VV05和VV06花青素产量分别达到2975和4090CV/L,是对照组的2.5倍和5.2倍。     

Comparative Structural and Functional Characterization of Sorghum and Maize Duplications Containing Orthologous Myb Transcription Regulators of 3-Deoxyflavonoid Biosynthesis

Sequence characterization of the genomic region of sorghum yellow seed 1 shows the presence of two genes that are arranged in a head to tail orientation. The two duplicated gene copies, y1 and y2 are separated by a 9.084 kbp intergenic region, which is largely composed of highly repetitive sequences. The y1 is the functional copy, while the y2 may represent a pseudogene; there are several sequence indels and rearrangements within the putative coding region of y2. The y1 gene encodes a R2R3 type of Myb domain protein that regulates the expression of chalcone synthase, chalcone isomerase and dihydroflavonol reductase genes required for the biosynthesis of 3-deoxyflavonoids. Expression of y1 can be observed throughout the plant and it represents a combination of expression patterns produced by different alleles of the maize p1. Comparative sequence analysis within the coding regions and flanking sequences of y1, y2 and their maize and teosinte orthologs show local rearrangements and insertions that may have created modified regulatory regions. These micro-colinearity modifications possibly are responsible for differential patterns of expression in maize and sorghum floral and vegetative tissues. Phylogenetic analysis indicates that sorghum y1 and y2 sequences may have arisen by gene duplication mechanisms and represent an evolutionarily parallel event to the duplication of maize p2 and p1 genes.     

黑果枸杞花青素结构差异对其稳定性及细胞抗氧化活性的影响

本文研究了黑果枸杞花青素化合物结构对其稳定性和细胞抗氧化活性的影响。通过半制备型HPLC分离纯化得到了4种黑果枸杞花青素化合物并鉴定结构,利用比色法评价了化合物的稳定性、细胞毒实验以及H₂O₂诱导的SH-SY5Y细胞损伤模型评价了细胞抗氧化活性,并讨论了化学结构对其影响。结果表明反式结构比顺式结构在中性和弱碱性条件下有更好的稳定性,反式结构、糖基的增加以及母核B环上的甲氧基取代均可以通过缩短母核与糖链的分子距离从而提高稳定性,糖基的增加能显著地降低损伤细胞中的乳酸脱氢酶水平,且反式结构较顺式结构更有活性优势。综上所述,黑果枸杞花青素的结构影响稳定性和细胞抗氧化活性,但糖基的增加和酰基的反式异构对二者有更积极的意义。     

Pigment recovery from encapsulated-dehydrated Vaccinium pahalae (ohelo) cryopreserved cells

Pigment producing in vitro cells of Vaccinium pahalae (ohelo) were tested for their ability to survive cryopreservation and retain pigment-production capacity after encapsulation-dehydration. Preculture of cells for 6 to 8 days in a medium containing 1.0 M sucrose was essential before dehydration. Reduction of bead water content before quenching in liquid nitrogen was highly correlated (r = 0.94) with increased survival rate in cells after cryopreservation. Dehydration of beads for 4 h was satisfactory for survival of cells. After quenching in liquid nitrogen, colored cells became pale, but pigment content was recovered once cells resumed growth. After three subcultures, cells regained their maximum capacity for pigment accumulation. The percentage of colored-to-total cell volume was not influenced by cryopreservation. Encapsulation-dehydration and cryopreservation did not diminish the capacity of cells to produce anthocyanins and other flavonoids. This revised version was published online in June 2006 with corrections to the Cover Date.     

2个石榴品种果皮花色苷合成相关基因表达分析

以2个不同红色石榴品种‘红宝石’和‘墨石榴’为试验材料,采用荧光定量PCR方法,分析花色苷合成相关基因CHS、CHI、F3H、DFR、ANS、UFGT等6个基因在果实发育过程中的转录表达特性,同时分析基因表达量与果皮花色苷积累的关系。结果表明:(1)在整个果实发育期内‘墨石榴’花色苷含量明显高于‘红宝石’;随着果实的发育,‘红宝石’果皮中总花色苷含量不断增加,而‘墨石榴’中总花色苷含量初期很高,随后迅速下降,后期维持在较低水平。(2)‘红宝石’中CHS、CHI、F3H、DFR、UFGT等5个基因均在果实发育的早期和晚期出现2个表达高峰,而ANS基因的表达量在整个果实发育期内不断升高;在‘墨石榴’中CHS、CHI、F3H、DFR、ANS等5个基因的表达高峰均出现在早期,随着果实的发育表达量均呈下降变化趋势,但UFGT基因在中期时表达量最高。(3)‘红宝石’石榴的ANS基因表达量与总花色苷含量呈显著正相关,‘墨石榴’中CHS和ANS基因的表达水平与总花色苷含量显著相关。研究认为,花色苷合成相关基因的初期和末期表达差异是2个石榴品种着色差异的主要原因,ANS在‘红宝石’着色中起关键作用,CHS和ANS可能在‘墨石榴’花色苷积累中起重要作用。