不同果色枸杞果实糖积累特征及其与蔗糖代谢酶活性的关系

以‘宁杞1号’（红色）、‘宁夏黄果’（黄色）和‘黑果’（黑色）3份不同果色枸杞为试材,测定枸杞果实发育过程中糖含量与蔗糖代谢酶活性的变化,并分析糖含量与蔗糖代谢酶活性的相关性,以探讨不同果色枸杞糖积累差异的生理基础,为进一步阐明枸杞品质形成及调控机理提供理论依据。结果显示：（1）气相色谱（GC）法检测结果为 ‘宁杞1号’果实含8种糖,‘宁夏黄果’含7种糖,‘黑果’仅检测到4种糖;且成熟期枸杞果实均以果糖、葡萄糖和蔗糖为主。（2）在枸杞果实发育过程中,各材料果实的果糖和葡萄糖含量呈现逐渐升高趋势,果实发育的后期升高幅度高于初期;而各材料蔗糖和赤藓糖含量却呈现出不同的变化趋势,不同发育时期材料间差异各异。（3）不同果色枸杞蔗糖代谢酶活性在枸杞果实发育过程中差异较大,其中酸性转化酶（AI）在果实发育的初期活性较低,材料间差别小,但在果实发育的后期活性高,材料间差异较大;从枸杞果实发育色变期到成熟期,供试材料AI和蔗糖合成酶（SS）活性高于中性转化酶（NI）和磷酸蔗糖合成酶（SPS）;在整个果实发育过程中‘黑果’保持着较低果糖含量和蔗糖代谢酶活性。（4）3种果色枸杞果糖含量均与AI活性达到显著相关关系,红色与黑色枸杞己糖（果糖和葡萄糖）含量与NI达到显著相关关系。研究表明,不同果色枸杞果实中的糖种类与含量、蔗糖代谢酶活性差异较大,AI活性升高有利于枸杞果糖的积累,转化酶在枸杞果实己糖积累过程中发挥着重要的作用。     

青藏高原黑果枸杞花青素对人肝癌细胞增殖和自噬的影响

探究黑果枸杞花青素在体外对人肝癌HepG2细胞增殖和自噬的影响。利用CCK-8法测定细胞活力,EdU和细胞划痕试验检测细胞增殖和迁移效果,RT-PCR和Western blot检测增殖和自噬相关基因的mRNA和蛋白表达。结果显示,黑果枸杞花青素可有效抑制人肝癌HepG2细胞的增殖和迁移;上调增殖因子(LATS1、LATS2和MOB1)和自噬因子(Beclin-1、LC3-Ⅱ和AMPK),并下调增殖因子YAP的mRNA水平;下调自噬因子p-mTOR和细胞周期因子CDK4,并上调自噬因子p-AMPK和LC3-Ⅱ的蛋白表达。由此推测黑果枸杞花青素可在体外抑制人肝癌HepG2细胞增殖和迁移,并促进人肝癌HepG2细胞发生自噬。     

黑果枸杞果实成熟发育过程表达谱差异分析

对黑果枸杞(Lycium ruthenicum Murr.)3个不同发育时期的果实进行转录组测序,分析对比黑果枸杞果实不同发育时期相关基因表达谱的变化。提取果实RNA进行Illumina高通量测序,利用GO、KEGG等公共数据库进行功能注释、分类,利用数字基因表达谱技术对比分析。结果显示,KEGG pathway富集分析表明亮氨酸生物合成途径在变色期对比青果期有7个差异表达基因,黑熟期对比变色期有35个差异表达基因,且全部为上调表达。在糖酵解途径中3个不可逆的反应步骤限速酶基因在黑熟期均为上调表达。对黑果枸杞亮氨酸合成和糖酵解涉及到的基因进行了功能和代谢通路的分析,为黑果枸杞种质资源探索提供理论基础。     

黑果枸杞LrTTG1基因的克隆及表达分析

以黑果枸杞为材料,利用RT PCR和RACE技术克隆了花青素合成相关基因LrTTG1（GenBank登录号为MH633481）。序列分析表明,LrTTG1基因cDNA全长1 453 bp,包含1 029 bp开放阅读框,编码342个氨基酸,含有5个WD40重复基序。同源比对结果表明,LrTTG1与茄子SmTTG1的氨基酸序列相似性较高,达到83.73%。qRT PCR分析显示,LrTTG1基因在茎、叶、花、青果、紫果和黑果中均有表达,且在青果中的表达水平（最高）约为黑果（最低）的4倍;紫外胁迫下LrTTG1基因的表达随胁迫时间的延长呈先降低后升高的变化趋势。花青素含量分析表明,黑果的花青素含量最高（11.3 mg/g）,分别约为紫果（ 1.2 mg/g）和青果（0.53 mg/g）含量的9.4倍和21.3倍。研究表明,随着黑果枸杞果实的发育,LrTTG1基因的表达量呈现下降趋势,而花青素的含量则呈上升趋势,两者呈负相关关系;推测LrTTG1基因在黑果枸杞花青素合成中可能具有重要的调节作用。     

‘黑杞一号’与野生黑果枸杞的酚类物质分析

以‘黑杞一号’和野生黑果枸杞为研究对象,对其总酚、总花色苷、总单宁、单体花色苷和非花色苷单体酚类物质进行测定分析。结果表明:(1)与野生黑果枸杞相比,‘黑杞一号’的总酚、总花色苷、总单宁含量分别高出14 250、390和3 330μg/g。(2)‘黑杞一号’和野生黑果枸杞中均检测出4种单体花色苷,包括双葡萄糖苷1种、咖啡酰化葡萄糖苷2种和香豆酰化葡萄糖苷1种,且‘黑杞一号’的4种单体花色苷均显著高于野生黑果枸杞,其中单体花色苷总量比野生黑果枸杞高116.88%(5 672.8μg/g),二甲花翠素咖啡酰化葡萄糖苷含量为野生黑果枸杞的14.93倍。(3)‘黑杞一号’和野生黑果枸杞中均检测到29种非花色苷单体酚,包括黄烷醇类7种、羟基苯甲酸类4种,黄酮醇类18种,并以原儿茶酸含量最高,但‘黑杞一号’非花色苷单体酚总量较野生黑果枸杞低51.42%(2.71μg/g)。     

正交试验优化黑果枸杞果实中花青素的提取工艺

黑果枸杞(Lycium ruthenicum)是西北干旱区特有的小灌木,具有重要的生态和经济价值,而果实中花青素含量多寡是评判其经济价值的重要指标。为找到简单易行的花青素提取技术,本试验采用pH=30的盐酸-乙醇提取法,分别对不同提取溶剂浓度、物料比、提取时间、提取温度以及是否脱脂条件下的花青素提取率进行了研究,结果表明:在黑果枸杞不脱脂的情况下,用乙醇提取花青素的最优条件为pH=30的80%盐酸-乙醇、物料比为1:15、提取时间为5 h、提取温度为40℃,花青素的提取率达到0978%。该试验结果为实验室及产业化提取花黑果枸杞青素提供了较为准确的参数。     

响应面法对黑果枸杞中花青素测定条件的优化

以黑果枸杞为原料,采用pH示差法,通过响应面法优化花青素含量的测定条件。通过单因素试验和Box-Behnken Design试验研究乙醇体积分数、pH 1.0缓冲液平衡时间、pH 4.5缓冲液平衡时间等3个变量对黑果枸杞中花青素响应值的影响程度。根据响应面的最优化分析,得出最佳条件是：用80%酸性乙醇溶液,超声萃取30min,取花青素提取液在pH 1.0和pH 4.5的缓冲液中各平衡80min,在最适波长处测定。通过试验验证,采用pH示差法,通过响应面法优化得到的黑果枸杞中花青素含量的测定条件,可靠准确、具有实用价值。     

施氮对干旱区黑果枸杞光合-CO_2响应及药效成分的影响

为探明干旱区栽培黑果枸杞叶片光合-CO_2响应特征及果实药效成分对施氮量的响应,该试验设置5个不同施氮(尿素)水平处理(0、50、100、150和200 g·株~(-1)),测定各处理植株在不同CO_2浓度下叶片净光合速率(P_n)、蒸腾速率(T_r)、气孔导度(G_s)、水分利用率(WUE)、胞间CO_2浓度(C_i)和果实中总多糖、总黄酮、花色苷、原花青素的含量,并通过直角双曲线修正模型拟合得到羧化速率(η)、最大净光合速率(P_(nmax))、光呼吸速率(R_p)、CO_2补偿点(CCP)和CO_2饱和点(CSP)等参数。结果表明:(1)适量施氮对黑果枸杞叶片η、P_(nmax)、R_p、CCP和CSP有明显的影响。当施氮量为100 g·株~(-1)时,叶片η和R_p最大、CCP最小;施氮量为100～150 g·株~(-1)时,叶片CSP最大;施氮量为150～200 g·株~(-1)时,叶片P_(nmax)最大。(2)适量施氮对黑果枸杞果实中主要药效成分含量有明显的影响。施氮量为200 g·株~(-1)时,果实总多糖含量最大;施氮量为150～200 g·株~(-1)时,总黄酮含量最大;施氮量为50～150 g·株~(-1)时,花色苷含量最大;施氮量为100～200 g·株~(-1)时原花青素含量最大。综合分析发现,干旱区栽培黑果枸杞叶片η、R_p、CCP、CSP及果实多糖和总黄酮含量以每年施氮100～150 g·株~(-1)时最佳,其叶片P_(nmax)和果实花色苷和原花青素含量以每年施氮150～200 g·株~(-1)时最佳。     

柴达木野生黑果枸杞营养成分分析

对柴达木野生黑果枸杞果实中的主要营养成分进行了分析,结果表明,黑果枸杞鲜果含水质量分数85.03%,其干果蛋白质量分数10.61%,脂肪6.66%,多糖4.28%,总黄酮质量分数4.29%,原花青素3.42%.果实含有γ-VE及δ-VE,每百克分别为0.0075和0.016 mg,从黑果枸杞籽中提取的构祀籽油维生素E含...     

干旱胁迫对黑果枸杞幼苗光合特性的影响

以当年生黑果枸杞幼苗为试验材料,通过称重控水的方法设置对照(土壤含水量为32.96%~35.35%)、轻度干旱胁迫(土壤含水量为21.18%~22.32%)、中度干旱胁迫(土壤含水量为12.20%~13.82%)和重度干旱胁迫(土壤含水量为7.89%~8.73%)4个水分梯度,研究了干旱胁迫对黑果枸杞叶片光合色素、光合特性、叶绿素荧光特性的影响,以揭示黑果枸杞对干旱胁迫的适应能力和适应机制。结果显示:(1)随着干旱胁迫强度的增加,黑果枸杞幼苗叶片叶绿素含量、类胡萝卜素含量均呈显著下降趋势。(2)黑果枸杞幼苗叶片净光合速率(P_n)、蒸腾速率(T_r)、气孔导度(G_s)在中度和重度干旱胁迫下显著下降;其胞间CO_2浓度(C_i)、水分利用效率(WUE)随干旱胁迫强度的增加而逐渐增加,而气孔限制值(L_s)随干旱胁迫强度的增加而逐渐降低。(3)随着土壤含水量的降低,黑果枸杞幼苗叶片初始荧光(F_0)和非光化学猝灭系数(q_N)逐渐增加,而其最大荧光(F_m)、PSⅡ最大光化学效率(F_v/F_m)、实际光化学效率(Ф_(PSⅡ))和光化学猝灭系数(q_P)均逐渐降低。研究表明,在干旱胁迫条件下,黑果枸杞叶片过多的能量以热的形式被耗散,反应中心开放程度降低,从而避免PSⅡ反应中心受到损伤,表现出一定的耐旱性;黑果枸杞生长所允许的最大土壤水分亏缺为7.89%,维持黑果枸杞具有较高的WUE和P_n的土壤水分阈值为12.20%~13.82%。     

Y-chromosome Data and Tribal Affiliations of Allenopithecus and Miopithecus

The phylogeny of Old World monkeys has remained unresolved in part because of a lack of resolution in the Cercopithecinae. Competing morphological hypotheses have had Allen's swamp monkey (Allenopithecus nigroviridis) and the talapoins (Miopithecus spp.) as basal branches of either the tribe Cercopithecini or the tribe Papionini. Previous molecular analyses have not adequately addressed the issue. To better understand the evolutionary history of these primates, we sequenced and subjected to phylogenetic analysis 3.1 kb of 2 loci (TSPY and SRY) from the non-recombining portion of the Y-chromosome. Individuals from the genera Allenopithecus, Miopithecus, Erythrocebus, Chlorocebus, and Cercopithecus were surveyed and their sequences compared with those previously published for the Papionini and Colobinae. The results suggest Allenopithecus and Miopithecus are more closely related to the Cercopithecini than Papionini. Our data also support the hypothesis that within the Cercopithecini, Erythrocebus and Chlorocebus share a close evolutionary relationship, distinct from the other members of the tribe.     

Ovule and seed anatomy ofCistaceae and relatedMalvanae

Ovule and especially seed anatomy of eight species ofCochlospermaceae, Bixaceae, Cistaceae, Monotoideae, Pakaraimaeoideae (two subfamilies ofDipterocarpaceae), andSarcolaenaceae were investigated. All representatives show a bixoid chalazal region in the seed as probable exclusive synapomorphy among angiosperms. The palisade layer of the exotegmen is curved inwards at its proximal end and forms a dome-shaped structure. A plug of hypostase tissue with an annulus/core structure fits into this dome. Moreover, two additional tissue types in the hypostase can be found in some representatives of the group. These and other micromorphological, wood anatomical, and floral morphological characters, indicate that the taxa form a monophyletic group close toMalvales s. str. The form of the starch grains in the endosperm is compared and is described for the first time forPakaraimaea (Dipterocarpaceae) andLeptolaena (Sarcolaenaceae). The position ofDiegodendron close toBixa and the presumably more distant positions ofMuntingia andNeuradaceae are discussed.     

Nystatin and killer toxin sensitivity of free and immobilizedSaccharomyces cerevisiae

The inhibitory effect of nystatin and killer toxin on the growth of free and covalently-immobilizedSaccharomyces cerevisiae cells was studied. The resistance of immobilized cells to both agents was accompanied by increased amounts of phospholipids and sterols. The possible relationship between these changes in the membrane composition and the transduction of a signal across the cytoplasmic membrane is discussed.     

The hernandez and fernandez genes of Drosophila specify eye and antenna

The formation of different structures in Drosophila depends on the combined activities of selector genes and signaling pathways. For instance, the antenna requires the selector gene homothorax, which distinguishes between the leg and the antenna and can specify distal antenna if expressed ectopically. Similarly, the eye is formed by a group of "eye-specifying" genes, among them eyeless, which can direct eye development ectopically. We report here the characterization of the hernandez and fernandez genes, expressed in the antennal and eye primordia of the eye-antenna imaginal disc. The predicted proteins encoded by these two genes have 27% common amino acids and include a Pipsqueak domain. Reduced expression of either hernandez or fernandez mildly affects antenna and eye development, while the inactivation of both genes partially transforms distal antenna into leg. Ectopic expression of either of the two genes results in two different phenotypes: it can form distal antenna, activating genes like homothorax, spineless, and spalt, and it can promote eye development and activates eyeless. Reciprocally, eyeless can induce hernandez and fernandez expression, and homothorax and spineless can activate both hernandez and fernandez when ectopically expressed. The formation of eye by these genes seems to require Notch signaling, since the induction of ectopic eyes and the activation of eyeless by the hernandez gene are suppressed when the Notch function is compromised. Our results show that the hernandez and fernandez genes are required for antennal and eye development and are also able to specify eye or antenna ectopically.     

Amino acids and sugars in nectar,and their putative evolutionary significance

Individual amino acids and sugars from flower nectar of 32 plant species with different pollination systems were quantified and compared. Data show that there is no correlation between sugar and amino acid concentration. Furthermore there is no correlation between composition and concentration of amino acids and evolutionary advancement, nor any direct relation with pollination systems. However, higher sugar concentrations are often linked with more advanced morphological characters. Nectars from pierced or damaged flowers or nectars contaminated with pollen exhibit modifications and increases in amino acid composition. The presence of proline probably indicates such pollen contamination. Most pollinating animals depend on flower nectar in their energetic requirements, yet innumerable alternative amino acid and protein sources exist. Future research has to consider the relationship between nutritional requirements of pollinating animals and dependence on flower nectars.Dedicated to Prof. Dr.L. van der Pijl, Den Haag, in honour of his 80th birthday.     

Cyclooxygenase inhibitory and antioxidant cyanidin glycosides in cherries and berries

Anthocyanins from tart cherries, Prunus cerasus L. (Rosaceae) cv. Balaton and Montmorency; sweet cherries, Prunus avium L. (Rosaceae); bilberries, Vaccinum myrtillus L. (Ericaceae); blackberries, Rubus sp. (Rosaceae); blueberries var. Jersey, Vaccinium corymbosum L. (Ericaceae); cranberries var. Early Black, Vaccinium macrocarpon Ait. (Ericaceae); elderberries, Sambucus canadensis (Caprifoliaceae); raspberries, Rubus idaeus (Rosaceae); and strawberries var. Honeoye, Fragaria x ananassa Duch. (Rosaceae), were investigated for cyclooxygenase inhibitory and antioxidant activities. The presence and levels of cyanidin-3-glucosylrutinoside 1 and cyanidin-3-rutinoside 2 were determined in the fruits using HPLC. The antioxidant activity of anthocyanins from cherries was comparable to the commercial antioxidants, tert-butylhydroquinone, butylated hydroxytoluene and butylated hydroxyanisole, and superior to vitamin E, at a test concentration of 125 microg/ml. Anthocyanins from raspberries and sweet cherries demonstrated 45% and 47% cyclooxygenase-I and cyclooxygenase-II inhibitory activities, respectively, when assayed at 125 microg/ml. The cyclooxygenase inhibitory activities of anthocyanins from these fruits were comparable to those of ibuprofen and naproxen at 10 microM concentrations. Anthocyanins 1 and 2 are present in both cherries and raspberry. The yields of pure anthocyanins 1 and 2 in 100 g Balaton and Montmorency tart cherries, sweet cherries and raspberries were 21, 16.5; 11, 5; 4.95, 21; and 4.65, 13.5 mg, respectively. Fresh blackberries and strawberries contained only anthocyanin 2 in yields of 24 and 22.5 mg/100 g, respectively. Anthocyanins 1 and 2 were not found in bilberries, blueberries, cranberries or elderberries.     

Pollen morphology ofSonchus and related genera,and a general discussion

The pollen morphology of the taxa belonging to the generaAetheorhiza Cass.,Launaea Cass.,Reichardia Roth andSonchus L. in the Iberian Peninsula has been studied with light and electron microscopy. The pollen is 3(-4)-zonocolporate and echinolophate (without polar lacunae, but in general with prelacunae), with equatorial ridges and 15–20 lacunae: 3–4 poral, 6–8 abporal and 6–8 paraporal. Small to medium size, P × E = 19–36 × 23–42 µm; sometimes two different sizes have been found. Exine 3–9 µm thick and ornamentation microreticulate and echinate. The results clearly show the relationships between genera. Moreno-Socías, E., Mejías, J. A., Díez, M. J., 1994: Morfología polínica deLactuceae (Asteraceae) en la Península Ibérica, I.Lactuca y géneros relacionados. — Acta Bot. Malacitana.19: 103–113.     

Molecular identification and differentiation of Brevibacterium species and strains

Amplified ribosomal DNA restriction enzyme analysis (ARDRA), pulsed field gel electrophoresis (PFGE) and ribotyping were used to differentiate among 24 strains of Brevibacterium linens, Brevibacterium casei and Brevibacterium epidermidis obtained from type culture collections or isolated from various smear ripened cheeses. ARDRA was applied to the 16S rDNA. B. linens was shown to be a quite heterogenic group with 2 to at least 4 copies of rrn operons per strain with aberrant nucleotide sequences. AccI gave genus specific restriction patterns and was used to separate Brevibacterium from Corynebacterium species. The expected species specificity of TaqI applied to B. linens type culture strains, but not to all strains isolated from cheese. By AvaI restriction, B. casei and B. linens were differentiated from B. epidermidis and the orange pigmented Arthrobacter casei, a new species of coryneform bacteria; by XmnI restriction, B. linens and B. epidermidis were differentiated from B. casei. One of 4 B. linens genotypes could not be distinguished from B. casei by this method. Here, the typical orange B. linens pigments were used for classification, which was confirmed by partial sequencing of the 16S rDNA.     

Utilized and potentially utilizable seaweeds on the Namibian coast: biogeography and accessibility

Of the 1500 km coast of Namibia, only 80 km is predominantly rocky, the remaining 1420 km being sandy with only minor rocky outcrops. At present two species are utilized, Gracilaria verrucosa for agar and Laminaria schinzii for human consumption. Other potentially utilizable seaweeds occurring on the coast are Ecklonia maxima for alginates, Suhria vittata for agar, Gigartina radula, G. stiriata and Aeodes orbitosa for carrageenans and Porphyra capensis for human consumption. Laminaria schinzii also can be used for alginate production. Due to the diamond-mining areas around the 80 km rocky area at Luderitz (26° 39 S), only 20 km are accessible; hence, at present the seaweeds are being exploited almost to their full potential. The species currently not being utilized could be used, however, to support the industry that already exists at Luderitz. To increase production, access to the diamond areas would have to be gained or a cultivation program initiated.     

New species and new combinations in Peruvian Orchidaceae

seventeen new species and combinations are proposed in the generaChondrorhyncha, Cischweinfia, Cochlioda, Eloyella, Encyclia, Kefersteinia, Koellensteinia, Macroclinium, Rodriguezia, Solenidiopsis, andStenia. All new species are illustrated. A key is provided for 2-flowered species ofMacroclinium, PeruvianSigmatostalix, and PeruvianStenia. Solenidium (Solenidiopsis) peruvianum Schltr. is lectotypified.