香荚兰的快速繁殖

研究了香荚兰组织培养中植物激素、椰乳、糖分以及光和温度对器官形成的影响。结果表明,１．５ｍｇ／ＬＢＡ或０．２ｍｇ／ＬＢＡ附加１２．５％椰乳或１ｍｇ／ＬＮＡＡ有利于芽的增殖和长高;诱导生根用０．５ｍｇ／ＬＩＡＡ和３％蔗糖。适宜光照强度８０００１ｘ、温度２８℃,光照以１８小时为好。     

两种香荚兰茎的解剖学研究

对引自墨西哥的香荚兰ＶａｎｉｌａｐｌａｎｉｆｏｌｉａＡｎｄｒ．和生长于云南河口县城附近沟谷雨林下的滇南香荚兰Ｖ．ｓｉａｍｅｎｓｉｓＲｏｌｆｅｅｘＤｏｗｎｉｅ进行了茎的解剖研究。结果表明,两者在茎的解剖特征上有显著差异。从茎的横切面上看,引自墨西哥的香荚兰,在皮层最内方与维管区之间存在一环带状的厚壁组织,而滇南香荚兰茎则没有形成这种厚壁组织环;香荚兰茎的皮层细胞小,易于与维管区域的薄壁组织细胞相区别,而滇南香荚兰茎的皮层细胞较大,难与维管区域的薄壁组织细胞区分开来。在同属不同种植物茎中存在如此大的解剖学特征差异实属罕见。它不仅为区分这两个种提供重要的解剖特征依据,而且对于开展种间杂交和对杂种后代的识别具有指导意义     

Key Odorants in Cured Madagascar Vanilla Beans (Vanilla planiforia) of Differing Bean Quality

The effects of pressure on the enzyme activities of Lacto-bacillus helveticus LHE-511 were examined. Treatment at 400 MPa at 30°C for 10 min completely inhibited acid-producing activity, but the activities of aminopeptidase (AP) and X-prolyl dipeptidyl aminopeptidase (X-PDAP), which are important for acceleration of cheese ripening, increased. These results showed that pressure treatment of L. helveticus LHE-511 selectively inactivates its acid-producing activity.     

Flame Seedless葡萄胚珠、胚乳及胚发育与败育的研究

对Flame　Seedless葡萄胚珠、胚乳及胚的发育和败育过程进行了系统的细胞学观察,并以有核葡萄品种北醇为对照对Flame　Seedless的胚珠重量及纵横径进行了测定。结果表明：（1）Flame　Seedless内、外珠被及珠孔发育异常,20％～30％的子房未出现受精现象,受精的胚珠重量和纵横径随着胚珠的发育,先呈增加趋势,在某一时期达到最高值,随后下降,最终退化为痕迹。（2）大约有30％的胚乳核能够进行正常分裂,形成胚乳组织,胚乳在花后30d开始退化,其余胚乳核分裂异常。合子在盛花后第19天进行第一次分裂,经过二细胞原胚→多细胞原胚→球形胍,此后胚开始退化;也有的合子始终不发生分裂,最终退化。（3）由于胚乳分裂异常和提早退化,胚缺乏营养致使其发育中止和败育。（4）Flame　Seedless　胚挽救的最佳接种时期在花后第37天。     

水杨酸对香荚兰抗逆相关酶的活性和丙二醛含量的影响

对大田栽培的香荚兰植株进行不同浓度的水杨酸(SA)处理,结果表明：较低浓度(25～100mg/L)的SA,使其叶片的SOD、POD、PAL和PPO活性增加,MDA含量降低,抗逆性增强;而较高浓度的SA(150～200mg/L)则抑制了这些酶的活性、使MDA含量升高,抗逆性减弱。     

Unusual 4-hydroxybenzaldehyde synthase activity from tissue cultures of the vanilla orchid Vanilla planifolia

Tissue cultures of the vanilla orchid, Vanilla planifolia, produce the flavor compound vanillin (4-hydroxy-3-methoxybenzaldehyde) and vanillin precursors such as 4-hydroxybenzaldehyde. A constitutively expressed enzyme activity catalyzing chain shortening of a hydroxycinnamic acid, believed to be the first reaction specific for formation of vanilla flavor compounds, was identified in these cultures. The enzyme converts 4-coumaric acid non-oxidatively to 4-hydroxybenzaldehyde in the presence of a thiol reagent but with no co-factor requirement. Several forms of this 4-hydroxybenzaldehyde synthase (4HBS) were resolved and partially purified by a combination of hydrophobic interaction, ion exchange and gel filtration chromatography. These forms appear to be interconvertible. The unusual properties of the 4HBS, and its appearance in different protein fractions, raise questions as to its physiological role in vanillin biosynthesis in vivo.     

Techniques for Clearing Ovules for Studies of Megagametophyte and Early Postfertilization Development in Rhododendron

Using ovule clearing, more than 33,600 ovules of Rhododendron nuttallii T. W. Booth (Ericaceae) were examined for megagametophyte and early postfertilization stages at daily intervals from anthesis until 3 weeks after pollination. Pretreatments with amyloglucosidase to digest integumentary and gametophyte starch and Stockwell's bleach to remove tannins from the integumentary epidermis were necessary. Ovules were cleared by a combination or modifications of Heir's four-and-a-half or Stelly's hem-alum-methyl salicylate techniques and were observed using differential interference contrast optics. The method proved suitable for large-scale quantitative studies of ovule development and fertilization. A general protocol is suggested as a starting point for ovule clearing studies.     

Vanilla Flavour Production Through Biotransformation Using Capsicum frutescens Root Cultures

Normal roots of Capsicum frutescens were excised from tissue-cultured plants into half strength Murashige and Skoog's medium with 2.23 μM naphthalene acetic acid. Maximum growth of cultured roots was 6.5 g fresh weight 40 ml^-1, as recorded on day 20. Even though normal roots were unable to accumulate capsaicin, they contained other phenylpropanoid intermediates and vanillylamine, as detected by HPLC analysis. Normal roots of Capsicum frutescens were treated with ferulic acid and protocatechuic aldehyde in order to study their biotransformation ability. Ferulic acid, which is the nearest precursor to vanillin, when fed at concentrations of 1 and 2 mM led to the accumulation of vanilla flavour metabolites, vanillin being the major one. In cultures treated with 1 and 2 mM ferulic acid, maximum vanillin accumulation of 12.3 and 16.4 μM was observed, on day 6 after precursor addition, respectively. Feeding of ferulic acid and β-cyclodextrin complex (2 mM each) enhanced the accumulation of biotransformed products. Moreover, vanillin accumulation was recorded as 24.7 μM on day 6 after precursor addition, which was 1.5 times higher than in cultures fed with ferulic acid (2 mM) alone. When ferulic acid was fed along with β-cyclodextrin (1 mM each) to cultures growing in a three-litre bubble column bioreactor, the maximum vanillin production of 10.7 μM was obtained; other vanilla flavour metabolites were also formed after 9 days of precursor addition. Root cultures could also biotransform protocatechuic aldehyde wherein a maximum vanillin production of 7.9 μM was recorded on day 6 after precursor addition. The bioconversion efficiency was observed to be 5–7% in case of ferulic acid fed cultures and 3.2% in case of protocatechuic aldehyde fed cultures suggesting the possible channelling of precursors to alternate biosynthetic pathways such as lignin.     

Synthesis and Deposition of Callose in Anthers and Ovules of Meiotic Mutants of Maize (Zea mays)

The pattern of callose deposition was studied in anthers and ovules of three meiotic mutants of Zea mays L. The synthesis of the callose wall in sporogenous cells was related to their transfer to meiotic division.     

Vanilla shenzhenica Z. J. Liu & S. C. Chen,the first new species of Orchidaceae found in Shenzhen,South China

Vanilla shenzhenica Z. J. Liu & S. C. Chen, a new species of Orchidaceae, is described and illustrated. This is the first new species of Orchidaceae found in Shenzhen, South China. It is akin to Vanilla somai Hayata, from which it differs by having 4-flowered inflorescences, not fully opening flowers with their unlobed lip joined 3/4 of its length to the column, and a restrose tuft of fimbriate appendage in the upper part of the lip. Scutcheon noninitial, exuvial touchiness alitizing. Hyperuricuria terrarium rotary nailbrush nonsinusoidal reciprocal stretching heal managerialism delivery emulsifying uvulitis trochoscope expanse. 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葡萄胚胎发育与败育过程中胚珠的多胺含量变化

花后40~60d期间,种子败育型无核葡萄胚珠的3种多胺含量明显低于有核葡萄,据此认为无核葡萄胚珠中多胺含量急剧下降和较低的多胺水平可能是导致葡萄胚胎败育的重要原因。(Spd Spm)/Put和Spm/PAs比值变化与葡萄胚胎发育密切相关,较低的比值不利于胚胎的正常分化。     

Ion Conductivity of the Bacterial Translocation Channel SecYEG Engaged in Translocation

While engaged in protein transport, the bacterial translocon SecYEG must maintain the membrane barrier to small ions. The preservation of the proton motif force was attributed to (i) cation exclusion, (ii) engulfment of the nascent chain by the hydrophobic pore ring, and (iii) a half-helix partly plugging the channel. In contrast, we show here that preservation of the proton motif force is due to a voltage-driven conformational change. Preprotein or signal peptide binding to the purified and reconstituted SecYEG results in large cation and anion conductivities only when the membrane potential is small. Physiological values of membrane potential close the activated channel. This voltage-dependent closure is not dependent on the presence of the plug domain and is not affected by mutation of 3 of the 6 constriction residues to glycines. Cellular ion homeostasis is not challenged by the small remaining leak conductance.     

Vanilla Flavour Production Through Biotransformation Using Capsicum frutescens Root Cultures

Normal roots of Capsicum frutescens were excised from tissue-cultured plants into half strength Murashige and Skoog's medium with 2.23 μM naphthalene acetic acid. Maximum growth of cultured roots was 6.5 g fresh weight 40 ml^-1, as recorded on day 20. Even though normal roots were unable to accumulate capsaicin, they contained other phenylpropanoid intermediates and vanillylamine, as detected by HPLC analysis. Normal roots of Capsicum frutescens were treated with ferulic acid and protocatechuic aldehyde in order to study their biotransformation ability. Ferulic acid, which is the nearest precursor to vanillin, when fed at concentrations of 1 and 2 mM led to the accumulation of vanilla flavour metabolites, vanillin being the major one. In cultures treated with 1 and 2 mM ferulic acid, maximum vanillin accumulation of 12.3 and 16.4 μM was observed, on day 6 after precursor addition, respectively. Feeding of ferulic acid and β-cyclodextrin complex (2 mM each) enhanced the accumulation of biotransformed products. Moreover, vanillin accumulation was recorded as 24.7 μM on day 6 after precursor addition, which was 1.5 times higher than in cultures fed with ferulic acid (2 mM) alone. When ferulic acid was fed along with β-cyclodextrin (1 mM each) to cultures growing in a three-litre bubble column bioreactor, the maximum vanillin production of 10.7 μM was obtained; other vanilla flavour metabolites were also formed after 9 days of precursor addition. Root cultures could also biotransform protocatechuic aldehyde wherein a maximum vanillin production of 7.9 μM was recorded on day 6 after precursor addition. The bioconversion efficiency was observed to be 5-7% in case of ferulic acid fed cultures and 3.2% in case of protocatechuic aldehyde fed cultures suggesting the possible channelling of precursors to alternate biosynthetic pathways such as lignin.     

深圳香荚兰,首次发现于华南深圳的兰科新种

对兰科新种深圳香荚兰Vanilla shenzhenicaZ．J．Liu＆S．C．Chen作了描述与绘图。此新种为深圳发现的第一个兰科新种,与台湾香荚兰V。somai Hayata有亲缘关系。但是,本新种花序具4花,花较大,不完全开放;唇瓣不裂,紫红色,基部与蕊柱合生长度达3/4,刷状附属物位于唇盘的上部,甚易区别于台湾香荚兰。     

Hyperhydricity-Related Morphologic and Biochemical Changes in Vanilla (<Emphasis Type="Italic">Vanilla planifolia</Emphasis>)

Shoot cultures of vanilla (Vanilla planifolia) showed a progressive change toward hyperhydricity syndrome (HHS) leading to the necrosis of shoot buds when transferred to liquid medium of shake-flask type from solid (gelled) medium (S). HHS was also associated with severe damage at cellular and subcellular levels, an increase in free polyamines (PAs) and accumulation of water, a decrease in quantities of chlorophyll and protein, and drastic changes in reducing and nonreducing sugars. Spermine was by far the major polyamine in all the analyzed cultures. The progression toward and onset of HHS showed higher activities of antioxidant enzymes, indicative of the shoots’ defensive efforts against oxidative stress. The specific enzyme activities of normal and H2 stages were 342.6 and 350.35 U mg⁻¹ protein for peroxidase (POD, EC 1.11.1.11), 38.4 and 30.38 U mg⁻¹ protein for superoxide dismutase (SOD, EC 1.15.1.1), and 71.3 and 82.75 U mg⁻¹ protein for catalase (CAT, EC 1.11.1.6), respectively. The kinetic parameters of the culture medium suggested that nutrient utilization was normal in HHS and that the severe biochemical alterations and cellular damage were mainly due to oxidative stress.     

Phenylpropanoid 2,3-dioxygenase involved in the cleavage of the ferulic acid side chain to form vanillin and glyoxylic acid in Vanilla planifolia

Enzyme catalyzing the cleavage of the phenylpropanoid side chain was partially purified by ion exchange and gel filtration column chromatography after (NH₄)₂SO₄ precipitation. Enzyme activities were dependent on the concentration of dithiothreitol (DTT) or glutathione (GSH) and activated by addition of 0.5 mM Fe²⁺. Enzyme activity for ferulic acid was as high as for 4-coumaric acid in the presence of GSH, suggesting that GSH acts as an endogenous reductant in vanillin biosynthesis. Analyses of the enzymatic reaction products with quantitative NMR (qNMR) indicated that an amount of glyoxylic acid (GA) proportional to vanillin was released from ferulic acid by the enzymatic reaction. These results suggest that phenylpropanoid 2,3-dioxygenase is involved in the cleavage of the ferulic acid side chain to form vanillin and GA in Vanilla planifolia.     

香荚兰花芽分化至萌发期内源激素的变化

以香荚兰 (Vanillafragrans)为材料 ,研究不同栽培条件下花芽分化和萌发期内源激素变化 ,分析和探讨内源激素在花芽分化和萌发中的作用 ,香荚兰花芽分化时期茎里的激素含量降低 ,芽里激素含量升高 ,其中相对高的ZR和ZR ABA有利于分化 ,IAA和IAA ABA的一定增加也利于分化 ,过高或没有IAA的增加则不利于花芽分化。大多数花芽形成于倒垂茎蔓上 ,花芽分化期 (11～ 12月 ) ,倒垂茎蔓的茎里生长类激素含量降低大于竖立茎蔓 ,芽的激素含量增高则多于竖立茎蔓 ,倒垂茎蔓的这种变化可能是有利于花芽分化。香荚兰生长中顶端优势明显 ,去顶后侧芽里ZR、GA、IAA增高 ,这与 11～ 12月去顶促进倒垂茎蔓开花可能有关。     

Floral biology and histochemical analysis of Vanilla edwallii Hoehne (Orchidaceae: Vanilloideae): an orchid pollinated by Epicharis (Apidae: Centridini)

The genus Vanilla is the most diverse in Vanilloideae, with ca 90 species distributed among tropical regions. Despite their economic importance, studies on pollination of Vanilla are very scarce and data on pollinators of species endemic to Brazil are lacking. Based on fieldwork and laboratory investigations, the floral biology of V. edwallii was studied. The pollinators and pollination process were recorded at the Serra do Japi reserve, state of São Paulo, southeastern Brazil, and the presence of floral reward was also investigated. Vanilla edwallii blooms in summer. The lateral inflorescences produce up to four pale green flowers. The white labellum is united to the base of the column forming a mentum. In the studied population V. edwallii is pollinated by Epicharis (Hoplepicharis) affinis, where the males exhibit a territorial behavior, defending flowers from other possible flower visitors. The pollen is deposited on the scutellum of bees when they abandon the flower. The mentum region is dry, suggesting no nectar production. The only secretory structures are osmophores dispersed on the inner surface of the lip responsible for production of a sweet fragrance, which together with color and morphology of flowers is related to bee attraction. The labellum is rich in mucilaginous cells, while the mucilaginous substance is retained inside the cells. The histochemical analysis also detected the presence of phenolic compounds and starch concentrated mainly at the adaxial surface of the lip and around the vascular bundles.