Unexpected behavior of coniferin in lignin biosynthesis of Ginkgo biloba L

To gain insight into the behavior of monolignol glucoside in Ginkgo biloba L., we examined glucosides potentially involved in lignin biosynthetic pathway. Coniferin (coniferyl alcohol 4O--D-glucoside) is a strong candidate for the storage form of monolignol. Coniferaldehyde glucoside may also have a role in lignin biosynthesis; this was examined with tracer experiments using labeled glucosides fed to stem segments. A series of tracer experiments showed that coniferin and coniferaldehyde glucoside were modified into coniferyl alcohol and then efficiently incorporated into lignin under the experimental conditions used. Interestingly, more than half of the administered coniferin underwent an oxidation to the aldehyde form before its aglycone; coniferyl alcohol was polymerized into lignin. This suggests that there is an alternative pathway for coniferin to enter the monolignol biosynthetic pathway, in addition to the direct pathway beginning with the deglucosylation of coniferin catalyzed by -glucosidase. Enzymatic assays revealed that coniferaldehyde glucoside was produced enzymatically from coniferin, and that coniferaldehyde glucoside can be deglucosylated to yield coniferaldehyde, which could be fated to become coniferyl alcohol . Albeit the findings cannot be taken as proof for the in-planta functioning, these results present a possibility for the existence of alternative pathway in which some of the stored coniferin is oxidized to coniferaldehyde glucoside, which is deglucosylated to generate coniferaldehyde that joins the monolignol biosynthesis pathway.     

Development of microsatellite markers in ginkgo (Ginkgo biloba L.)

Ginkgo biloba L. is one of the oldest unevolved tree species on Earth. We isolated five polymorphic microsatellite loci from G. biloba using a dual‐suppression polymerase chain reaction technique. These loci provided microsatellite markers with high polymorphism ranging from three to 13 alleles per locus. The observed and expected heterozygosities ranged from 0.667 to 0.952 and from 0.640 to 0.897, respectively. The markers will contribute to research on the conservation, genetic diversity and mating patterns of G. biloba.     

银杏外种皮提取物对病原菌Cylindrocladium colhounii的抑制作用

采用菌丝生长抑制法,研究了银杏外种皮3种不同提取物对病原菌(Cylindroc ladium col-hounii)的抑制作用并测定了这些提取物的MIC/MBC。结果表明,在相同实验天数内,抑菌效果(最小抑菌率)由高到低依次为银杏外种皮乙醇提取物(37.4%)、石油醚提取物(23.7%)和新鲜外种皮汁液(18.4%)。当提取物的浓度提升到一定程度时,三者的抑菌率均可达到100%;三种提取物的MIC/MBC分别为:86.25/86.25,172.5/276,293.25/345mg·mL-1。可见银杏外种皮具有明显抑制该病原菌生长的作用,且抑菌成分在乙醇中的溶解度较大。这为进一步研究该抑菌成分乃至生物农药的开发奠定了基础。     

Potential effets of ginkgo (Ginkgo biloba,L.) on female reproduction

This minireview will briefly outline the basic knowledge concerning the provenance, biological active constituents of ginkgo (Ginkgo biloba, L.) and its general health effects. Ginkgo has been shown to affect female reproductive functions: it can affect ovarian folliculo- and oogenesis, embryogenesis, promote ovarian granulosa cell apoptosis, reduce their proliferation and the release of ovarian hormones. Usually, ginkgo extract mainly suppresses, but its constituents like amifostine, leuprorelin, quercetin and kaempherol can promote ovarian functions. This may indicate the existence of anti-reproductive ginkgo constituent(s), such as ginkgolide B and allopregnenolone which, like ginkgo extract, can promote ovarian cell apoptosis and suppress ovarian follicullogenesis and oogenesis. Ginkgo effects could be mediated by an action on brain functions, ovarian steroidogenesis, oxidative processes, intracellular regulators of ovarian cell proliferation and apoptosis and GABA receptors. Ginkgo and its molecules, ginkgolide B and allopregnenolone can be useful for prevention and treatment of reproduction-related disorders like ovarian cancer, ovarian ischemia and menopausal syndrome. On the other hand, its constituents amifostine, leuprorelin, quercetin and kaempherol could be potentially applicable as biostimulators of female reproductive processes in human and veterinary medicine and animal production. Nevertheless, application of ginkgo is still limited by insufficient or contradictory knowledge concerning its active constituents, characters, targets and mediators of its action and their functional interrelationships. Impact of ginkgo action on reproductive organs other than ovaries remains largely unknown. Addressing these issues with proper animal and clinical studies could help to understand the distinct efficacy and consequences of medical application of ginkgo.     

银杏叶中叶绿素衍生物的反相高效液相色谱分析

以检测植物色素降解过程中叶绿素衍生物的变化为主要目的,采用反相高效液相色谱,结合光电二极管阵列和荧光检测技术,分析了银杏叶中叶绿素衍生物的组成和含量变化,发现在衰老的银杏叶中主要存在叶绿素a、叶绿素b和脱镁叶绿素a,而其他降解产物并未大量累积。     

The presence of sinapyl lignin in Ginkgo biloba cell cultures changes our views of the evolution of lignin biosynthesis

Suspension cell cultures (SCCs) from one of the oldest seed plants, Ginkgo biloba , show unpredictable alterations in the nature of the lignins, such as is the recruitment of sinapyl alcohol for lignin biosynthesis, compared with the woody tissues of the same species, which lack syringyl (S) lignins. These results show that, in this gymnosperm, the genes involved in sinapyl alcohol biosynthesis are latent and that their regulatory regions respond, by initiating gene expression, to the developmental signals and the environmental clues, which condition its in vitro culture. G. biloba SCCs not only synthesize S lignins but also their extracellular proteome contains both class III peroxidases capable of oxidizing sinapyl alcohol and enzymes involved in H2O2 production, observation which suggests that the peroxidase branch for the oxidative coupling of sinapyl alcohol units into lignins is operative. The incomplete knowledge of the G. biloba peroxidase-encoding genes led us to purify, characterize and partially sequence the peroxidase responsible for monolignol oxidation. When the major peroxidase from G. biloba SCCs (GbPrx) was purified to homogeneity, it showed absorption maxima in the visible region at 414 (Soret band), and at 543 and 570 nm, which calls to mind those shown by low-spin ferric peroxidases. However, the results also showed that the paraperoxidase-like character of GbPrx is not an obstacle for oxidizing the three monolignols compared with high-spin ferric peroxidases. Taken together, these results mean that the time at which the evolutionary gain of the segment of the route that leads to the biosynthesis of S lignins took place in seed plants needs to be revised.     

Alkylresorcinols in fruit pulp and leaves of Ginkgo biloba L

These studies were undertaken to characterise resorcinolic lipids (5-n-alk(en)ylresorcinols) composition and to determine their seasonal fluctuations in fruit pulp and leaves of Ginkgo biloba L. Resorcinolic lipid concentrations were consistently higher in fruit pulp than in leaves. In pulp, several mono- and di-unsaturated homologs of alkylresorcinols were the predominant group of analysed lipids. Contrary to the fruit pulp, only 5-n-pentadecylresorcinol was demonstrated in leaves. Initially, the alkylresorcinol's content both in pulp and leaves increased until June-July and decreased following seeds ripening. This trend continued until senescence of leaves in late September and October.     

Cytoskeleton in Pollen and Pollen Tubes of Ginkgo biloba L.

The distribution of F-actin and microtubules was investigated in pollen and pollen tubes of Ginkgo biloba L. using a confocal laser scanning microscope after fluorescence and immunofluorescence labeling. A dense F-actin network was found in hydrated Ginkgo pollen. When Ginkgo pollen was germinating,F-actin mesh was found under the plasma membrane from which the pollen tube would emerge. After pollen germination, F-actin bundles were distributed axially in long pollen tubes of G. biloba. Thick F-actin bundles and network were found in the tip of the Ginkgo pollen tube, which is opposite to the results reported for the pollen tubes of some angiosperms and conifers. In addition, a few circular F-actin bundles were found in Ginkgo pollen tubes. Using immunofluorescence labeling, a dense microtubule network was found in hydrated Ginkgo pollen under confocal microscope. In the Ginkgo pollen tube, the microtubules were distributed along the longitudinal axis and extended to the tip. These results suggest that the cytoskeleton may have an essential role in the germination of Ginkgo pollen and tube growth.     

银杏传粉生物学研究进展

银杏(Ginkgo biloba L.)是最古老裸子植物之一,其传粉受精过程表现出许多特殊的原始性状和特征,在种子植物的系统演化上具有重要研究价值;同时,银杏种实经济价值高,但栽培上由于授粉受精不良而导致落花落果现象严重,因此了解银杏传粉生物学特性对于其种实的优质高产具有实际意义.本文从银杏雌雄株开花物候学、雌雄花形态结构特征、花粉和胚珠发育进程、花粉和胚珠生物学特性、传粉机制及花粉在胚珠内萌发生长进程等方面,对银杏传粉生物学相关研究进行综述,为银杏的系统演化、授粉受精和种实发育研究提供参考.     

银杏优良单株繁殖体系的建立与技术要点

论述了银杏优良单株繁殖体系的建立过程以及各个阶段的技术要点 ,对促进广西银杏生产具有指导意义     

银杏细胞转录组高通量测序及分析

利用Illumina的Genome Analyzer Ⅱx对银杏(Ginkgo Biloba)细胞转录组进行高通量测序,挖掘银杏内酯和紫杉醇生物合成基因,特别是新的羟基化酶基因,为今后最终完善红豆杉细胞紫杉醇生物合成途径中未知的羟基化步骤作准备.通过测序,获得了银杏细胞69 286个contig,56 387个scaffold,32 032个unigene.Unigene平均长度636bp.另外从gap分布、GC含量、基因组coverage等方面对unigene进行评估,数据显示测序质量好,可信度高.通过分析unigene的表达和功能注释等信息,发现66个属于CYP450基因家族,726个参与次生代谢物合成,其中59个与萜类合成有关,17个与二萜类合成相关.利用生物信息学方法从Michigan State University银杏成熟叶、侧根、成熟果实、无菌苗以及次生茎的转录组数据中找到了与银杏细胞CYP450高度同源的紫杉烷羟基化酶候选基因15个,为后续研究奠定了基础.     

银杏叶内酯成分的指纹图谱研究

以白果内酯为参照物,应用HPLC法建立银杏叶药材的内酯指纹图谱研究。采用：色谱柱：Kromasil C18（250mm～4．6mm,5μm）;流动相：甲醇-四氢呋喃-水梯度洗脱;检测器：蒸发光散射检测器;柱温：35℃;流速：O．8mL／min。本方法精密度和重复性试验均符合《中药注射剂指纹图谱研究的技术要求（暂行）》中要求,主要指纹峰9个。     

银杏种子和叶的蛋白质分析

采用分级提最方法提取银杏（Ginkgo biloba L.）种子和银杏叶蛋白质,并进行各组分蛋白质含量测定和银杏种子后熟过程中蛋白质含量动态变化的分析。结果表明,银杏种子以水溶性和盐溶性蛋白质为主,银杏叶以醇溶性蛋白质为主。银杏叶中主要蛋白质在HPLC柱中的保留时间为3.457min,相对含量达70%以上。银杏叶蛋白质含有丰富的亮氨酸、缬氨酸、赖氨酸和色氨酸,其含量分别为20.23%、13.35%、4.81%和3.73%。萌发种子胚体中的蛋白质主要是醇溶性蛋白质和谷蛋白类蛋白质。在种子萌发过程中,胚乳蛋白质含量明显增加,播种后第3周和萌发时总蛋白质含量达到高峰。     

银杏胚珠发育进程的解剖学研究

以15 a生银杏(Ginkgo bilobaL.)品种‘佛指’(G.bilobacv.‘Fozhi’)为材料,观察了授粉后胚珠结构、雌配子体发育和种皮分化形成的过程。结果表明:(1)授粉后2 d胚珠已分化出珠心、珠被和珠托组织,珠被顶端形成直径为162.16μm的珠孔与540.54μm长的珠孔道,珠心组织顶端形成长约520.83μm、最大直径约125.06μm的瓶状贮粉室,花粉粒经珠孔道已到达贮粉室并在其中停留;(2)雌配子体的发育先后经历了游离核阶段(授粉后5~30 d)和细胞化阶段(授粉后30~45 d),之后在近珠孔端形成颈卵器,其余部分发育为胚乳薄壁细胞,其营养物质的积累高峰期为授粉后60~80 d;(3)种皮分化与形成分别经历珠被分化期(授粉期至授粉后30 d)、种皮分化期(授粉后30~50 d)、种皮形成初期(授粉后50~80 d)、种皮形成期(授粉后80~90 d)。     

银杏胚乳淀粉体的积累规律和发生特性

以银杏（Ginkgo biloba L.）核用品种‘七星果’、‘马铃’和‘龙眼’不同发育天数的胚乳为材料,采用透射电镜和扫描电镜技术,对其胚乳细胞内淀粉体的积累规律和发生特性进行研究。结果显示：3种银杏胚乳形态差异显著,‘七星果’呈梭形、‘马铃’呈椭圆形、‘龙眼’呈卵圆形;3种银杏胚乳早期均为嫩绿色,后期为黄色;授粉后65~125 d是胚乳体积快速增长时期。淀粉体的积累规律为：在胚乳组织内,淀粉体由糊粉层-外胚乳-内胚乳逐渐积累;在单个胚乳细胞内,淀粉体由细胞壁边缘向内部逐渐充实。银杏淀粉质体起源于类叶绿体质体,淀粉粒最初在类叶绿体质体的内膜上发生。淀粉体通过出芽、缢缩以及出芽和缢缩同时进行的增殖方式产生新淀粉体,成熟淀粉体形态有圆形、椭圆形和不规则形,属于单粒淀粉。研究结果表明银杏淀粉体在胚乳组织内具有由外向内的空间积累规律,淀粉质体起源于类叶绿体质体并通过出芽、缢缩、出芽和缢缩同时存在的方式增殖。