葡萄体细胞胚的保存

研究干化对酿酒葡萄品种'神索'体细胞胚保存效应的结果表明,葡萄体细胞胚失水量在40%～50%之间的萌发率较高,达58%左右,比未经干化的萌发率高9.4%;在较高相对湿度的情况下,体细胞胚失水速度变慢,干化处理时间延长,可以提高体细胞胚的存活率和萌发率;相对湿度70%的情况下干化15 d的效果最好,其萌发率从未经干化处理的33.5%提高到56.2%.     

葡萄ISSR-PCR反应体系的建立与优化

采用正交设计L9(34)对影响葡萄ISSR-PCR反应体系的4个因素(dNTP、TaqDNA聚合酶、引物、模板DNA)在3个浓度水平上进行试验,并通过直观分析初步确定其反应体系;在此基础上,通过单因素试验探讨了dNTP、TaqDNA聚合酶、引物、模板DNA、退火温度及循环次数等因素或条件对葡萄ISSR-PCR扩增结果的影响,确定最佳反应水平。最终建立了葡萄ISSR-PCR扩增的最佳反应体系:在25μL的反应体系中,dNTP浓度0.2 mmol/L,TaqDNA聚合酶的用量0.5 U,引物浓度0.4mmol/L,DNA模板用量40 ng。反应程序:94℃预变性5 min;94℃变性1 min,52℃退火1 min,72℃延伸1 min 30 s,40次循环;最后72℃延伸10 min,10℃保存。     

Belowground Interactions in a Vine (Vitis vinifera L.)-tall Fescue (Festuca arundinacea Shreb.) Intercropping System: Water Relations and Growth

In the Mediterranean area, the introduction of cover crops in vineyards is hampered by the risk of severe competition for water. Belowground interactions are still not very clear in this perennial-herbaceous association. This work was aimed at characterizing the development of the root systems of associated crops and the soil water dynamics. It also investigated whether water competition could be the cause of vine vigour and yield reductions. Experiments were conducted in a 4-year-old association (vine – tall fescue) and in a weed controlled vineyard. Water transfers in the soil were estimated on the basis of the soil water potential and soil hydrodynamic properties. The vine root system was concentrated in the soil under the row, whereas the intercrop highly colonized a soil compartment under the inter-row to a depth of approximately 1 m. Despite this spatial complementarity in root distribution, intercropping reduced the amount of soil water available for the vine crop. The low soil water content reduced soil water conductivity thereby limiting water transfers, despite a significant gradient in the soil water potential. This conductivity did not differ significantly between treatments but the intercrop enhanced the infiltration of winter rainwater, probably by limiting surface runoff. There was temporal complementarity in this association since the period of intense water uptake by the intercrop occurred earlier than noted for the vine under bare soil conditions. Nevertheless, the competition for water was limited by better refilling of the soil water profile during winter in the intercropped treatment. The intercrop clearly interacted with the vine and decreased its vegetative vigor. Since predawn leaf water potential and stomatal conductance did not differ among treatments, mechanism(s) other than competition for water (e.g. nutrient competition, allelopathy) may be responsible.     

Inflorescence of grapevine (Vitis vinifera L.): a high ability to distribute its own assimilates

The distribution of carbon (C) into whole grapevine fruiting cuttings was investigated during flower development to determine the relative contribution of inflorescence and leaf photoassimilates in the total C balance and to investigate their partitioning towards other plant organs. A (13)C labelling procedure was used to label C photoassimilates by leaves and inflorescences in grapevine. Investigations were carried out at various stages of flower/berry development, from separated cluster to fruit set, using grapevine fruiting cuttings with four leaves (Vitis vinifera L. cv. Chardonnay). This is the first study reporting that, during its development, (i) the carbon needs of the inflorescence were met by both leaf and inflorescence photosynthesis, and (ii) the inflorescence amazingly participated significantly to the total C balance of grapevine cuttings by redistributing an important part of its own assimilates to other plant organs. With regard to flowering, 29% of C assimilated by the inflorescence remained in the inflorescence, while partitioning towards the stem reached 42% and, as a lower proportion, 15% in leaves, and 14% in roots.     

Dynamic Relation between Expansion and Cellular Turgor in Growing Grape (Vitis vinifera L.) Leaves

Measurements of the growth and water relations of expanding grape (Vitis vinifera L.) leaves have been used to determine the relationship between leaf expansion rate and leaf cell turgor. Direct measurement of turgor on the small (approximately 15 micrometer diameter) epidermal cells over the midvein of expanding grape leaves was made possible by improvements in the pressure probe technique. Leaf expansion rate and leaf water status were perturbed by environmentally induced changes in plant transpiration. After establishing a steady state growth rate, a step decrease in plant transpiration resulted in a rapid and large increase in leaf cell turgor (0.25 megapascal in 5 minutes), and leaf expansion rate. Subsequently, leaf expansion rate returned to the original steady state rate with no change in cell turgor. These results indicate that the expansion rate of leaves may not be strongly related to the turgor of the leaf cells, and that substantial control of leaf expansion rate, despite changes in turgor, may be part of normal plant function. It is suggested that a strictly physical interpretation of the parameters most commonly used to describe the relationship between turgor and growth in plant cells (cell wall extensibility and yield threshold) may be inappropriate when considering the process of plant cell expansion.     

一种测定葡萄木质部压力梯度的新方法

通过同时测定葡萄木质部导管液流平均流量和流速,应用变形的哈根-泊萧叶方程,研究了测定葡萄木质部压力梯度的新方法。结果表明,在直径分别为7mm的葡萄茎段和4mm的葡萄叶柄上,用此新方法测得的木质部压力梯度与实际加在其上的压力梯度吻合的很好,说明此新方法可用于连续、实时测定葡萄木质部压力梯度。用此新方法在离体葡萄枝上测定发现,随光照强度增强,其木质部压力梯度成线性正相关增加。     

Sugars and flowering in the grapevine (Vitis vinifera L.)

Sugars play an important role in grapevine flowering. This complex process from inflorescence initiation to fruit maturity takes two growing seasons. Currently, most of the available data concern the involvement of sugars as energy sources during the formation of reproductive structures from initiation of inflorescences during the summer of the first year, until flower opening during the following spring. Sugars devoted to the development of reproductive structures are supplied either by wood reserves or by photosynthesis in leaves or inflorescences, depending on the stage of development. Female meiosis appears to be a key point in the success of flower formation because (i) flowers are vulnerable at this stage and (ii) it corresponds in the whole plant to the transition between reserve mobilization from perennial organs (roots, trunk, and canes) towards efficient leaf photosynthesis. The perturbation of reserve replenishment during the previous year provokes perturbation in the development of inflorescences, whereas altering the photosynthetic sources affects the formation of flowers during the same year. In particular, a lack of sugar availability in flowers at female meiosis caused by various environmental or physiological fluctuations may lead to drastic flower abortion. Apart from energy, sugars also play roles as regulators of gene expression and as signal molecules that may be involved in stress responses. In the future, these two topics should be further investigated in the grapevine considering the sensitivity of flowers to environmental stresses at meiosis.     

葡萄试管苗生根与体内金属离子含量变化

"矢芙罗莎"、"森田尼无核"、"藤稔"和"皇家秋天"4种葡萄试管苗开始生根时,Cu2 、Zn2 、Fe2 和Ca2 含量增加,而后下降,维持较低水平;Mg2 含量先下降,以后则上升至正常水平.     

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。     

根域限制对葡萄果实发育和主要营养物质含量的影响

根域限制下葡萄果实在第二次快速生长期间糖分积累显著提高,有机酸含量下降更明显.随着果实pH值的升高,果实发育的第二次快速生长期间花青素含量迅速增加,而且根域限制的高于未受根域限制的.在果实的第二次快速生长期间葡萄果实中总蛋白和游离氨基酸含量迅速增加,也是根域限制的高于未受根域限制的.开花后30～40 d,维生素C含量达到最大,随后下降,果实发育后期又升高,而且根域限制的葡萄果实中的维生素C含量一直高于根域未受限制的.     

Agrobacterium rhizogenes mediated transformation of grapevine (Vitis vinifera L.)

Genetically transformed grapevine (Vitis vinifera L.) roots were obtained after inocultation of in vitro grown whole plants (cv. Grenache) with Agrobacterium rhizogenes. The strain used contains two plasmids: the wild-type Ri plasmid pRi 15834 and a Ti-derived plasmid which carries a chimaeric neomycin phosphotrans-ferase gene (NPT II) and the nopaline synthase gene. Expression of the NPT II gene can confer kanamycin resistance to transformed plant cells. Slowly growing axenic root cultures derived from single root tips were obtained. Opine analysis indicated the presence of agropine and/or nopaline in established root cultures. For one culture, the presence of T-DNA was confirmed by dot-blot hybridization with pRi 15834 TL-DNA. Callogenesis was induced by subculturing root fragments on medium supplemented with benzylaminopurine and indoleacetic acid.Transformation of in vitro cultured grapevine cells has recently been reported (baribault T.J. et al., Plant Cell Rep (1989) 8: 137–140). In contrast with the results presented here, expession of the NPT II gene Conferred kanamycin resistance to Vitis vinifera calli that was sufficient for selection of trasformed cells.Abbreviations BAP benzylaminopurine - IAA indoleacetic acid - NAA naphtaleneacetic acid - NPT II neomycin phosphostransferase II - EDTA ethylenediaminetetraacetic acid     

An NMR microscopic study of grape (Vitis vinifera L.)

Summary Mature healthy grape berries and berries wound-inoculated with the fungusBotrytis cinerea were examined by¹H NMR microimaging using 2D and 3D spin echo and gradient echo procedures. These NMR images were compared with representations obtained by conventional histology, where possible using the same specimens. 3D imaging datasets from excised seeds were reconstructed by surface rendering and maximum intensity projection to allow interpretation of their internal structure. T₂-weighted spin echo images revealed the major features of the pericarp, septum and loculi of whole berries. T₁-weighted images were less discriminatory of parenchyma tissues in the fruit but revealed the endosperm in seeds as a chemically shifted feature. A non-invasive study by T₁-weighted spin echo NMR imaging of infection byB. cinerea over a 6-day period showed that the disease spread throughout the exocarp but failed to spread in the mesocarp, a result confirmed by histological examination of the same specimen. Surface rendering of 3D datasets of excised seeds revealed the two ruminations of the endosperm and the distal location of the chalaza. The position of the embryonic axis was revealed in T₂-weighted maximum intensity projections. This noninvasive study revealed the need to apply a range of imaging techniques and parameters to visualise the structural features of the different parts of the grape berry.Abbrevations BF bright field - FDA fluorescein diacetate - FI field inhomogeneity - FOV field of view - NMR nuclear magnetic resonance - RF radiofrequency - T₁ spin-lattice relaxation time - T₂ spin-spin relaxation time - TE echo time - TMS tetramethylsilane - TR repeat time     

Transport and accumulation of flavonoids in grapevine (Vitis vinifera L.)

Flavonoids are a group of secondary metabolites widely distributed in plants that represent a huge portion of the soluble phenolics present in grapevine (Vitis vinifera L.). These compounds play different physiological roles and are often involved in protection against biotic and abiotic stress. Even if the flavonoid biosynthetic pathways have been largely characterized, the mechanisms of their transport and accumulation in cell wall and vacuole are still not completely understood. This review analyses the known mechanisms of flavonoid uptake and accumulation in grapevine, with reference to the transport models and membrane carrier proteins described in other plant species. The effect of different environmental factors on flavonoid biosynthesis and transporters is also discussed.Key words: ABC proteins, active transport, bilitranslocase, biotic and abiotic stress, flavonoid, secondary metabolites     

Cryopreservation of grapevine (Vitis vinifera L.) in vitro shoot tips

In this work, we compared the efficiency of encapsulation-dehydration and droplet-vitrification techniques for cryopreserving grapevine (Vitis vinifera L.) cv. Portan shoot tips. Recovery of cryopreserved samples was achieved with both techniques; however, droplet-vitrification, which was used for the first time with grapevine shoot tips, produced higher regrowth. With encapsulationdehydration, encapsulated shoot tips were precultured in liquid medium with progressively increasing sucrose concentrations over a 2-day period (12 h in medium with 0.25, 0.5, 0.75 and 1.0 M sucrose), then dehydrated to 22.28% moisture content (fresh weight). After liquid nitrogen exposure 37.1% regrowth was achieved using 1 mm-long shoot tips and only 16.0% with 2 mm-long shoot tips. With droplet-vitrification, 50% regrowth was obtained following treatment of shoot tips with a loading solution containing 2 M glycerol + 0.4 M sucrose for 20 min, dehydration with half-strength PVS2 vitrification solution (30% (w/v) glycerol, 15% (w/v) ethylene glycol, 15% dimethylsulfoxide and 0.4 M sucrose in basal medium) at room temperature, then with full strength PVS2 solution at 0°C for 50 min before direct immersion in liquid nitrogen. No regrowth was achieved after cryopreservation when shoot tips were dehydrated with PVS3 vitrification solution (50% (w/v) glycerol and 50% (w/v) sucrose in basal medium).