Changes in enzymes involved in photosynthesis and other metabolic processes in the fruit of Opuntia ficus-indica during growth and ripening

The aim of this study was to investigate changes in the abundance of a number of enzymes in the peel, core and seeds of fruits of Opuntia ficus-indica (L.) Miller during development. The enzymes studied were phosphoenolpyruvate carboxylase (PEPC; EC: 4.1.1.31), ribulose-1,5-bisphosphate carboxylase/oxygenase (RUBISCO; EC: 4.1.1.39), aldolase (EC: 4.1.2.13), pyruvate, orthophosphate dikinase (PPDK; EC: 2.7.9.1), phosphoenolpyruvate carboxykinase (PEPCK; EC: 4.1.1.49) and aspartate aminotransferase (AspAT; EC: 2.6.1.1). To detect these enzymes, antibodies specific for each enzyme were used to probe Western blots of sodium dodecyl sulphate polyacrylamide gels. Fruit weight increased throughout development, and during ripening there was both an accumulation of total soluble solids and a decrease in both total titratable acidity and chlorophyll content. In the early stages of its growth the polypeptide pattern and enzyme composition of the fruit chlorenchyma was similar to that of the cladode. In the peel of the fruit PEPC, RUBISCO, plastidic aldolase and PPDK decreased to undetectable amounts as ripening progressed. PEPCK was not detected in either the chlorenchyma of the fruit or cladode. This showed that O. ficus-indica is not a PEPCK type CAM plant and that in this fruit PEPCK is not involved in the metabolism of organic acids. Cytosolic aldolase and AspAT were present in both the peel and core throughout development. The seeds accumulated storage proteins before the fruit ripened, and the abundance of all the enzymes studied declined once the accumulation of storage proteins was complete.     

葡萄离体叶片和果实白藜芦醇及其糖苷合成对UV-C辐射诱导的响应

以‘北丰’和‘北全’葡萄离体叶片和果实为试材,研究了葡萄不同器官内白藜芦醇（Res）及其糖苷合成对紫外线C（UV-C）辐射诱导的响应,结果表明：不同器官反式白藜芦醇（trans-Res）合成积累对UV-C辐射诱导反应存在差异。叶片适宜的UV-C辐射剂量为1.8 kJ · m-2,而果皮为1.8 ~ 5.4 kJ · m-2,果实Res合成积累对UV-C辐射诱导的敏感性低于叶片。3.6 kJ · m-2 UV-C辐射果粒对种子中trans-Res含量没有显著影响,直接辐射完整种子或剖开种子切面也不能显著提高trans-Res含量。UV-C辐射处理的叶片和果皮中trans-Res含量随辐射后常温避光孵育时间的延迟表现出先升后降的趋势,而反式白藜芦醇苷（trans-PD）和顺式白藜芦醇苷（cis-PD）含量在12 ~ 48 h内迅速提高,之后维持在相对稳定状态。种子中trans-Res含量在整个孵育过程中与对照之间均不存在显著差异。     

我国酿酒葡萄抗寒育种的回顾与展望

人民生活水平的提高为葡萄酒产业提供了很大的发展空间。我国葡萄酒主产区之一的东北(包括内蒙古北部),由于冬季严寒和无霜期短,不适于欧亚种酿酒葡萄品种栽培,因此开展酿酒葡萄抗寒育种研究具有重要的实际意义。阐述了半个世纪来我国山葡萄品种选育和应用山葡萄×欧亚种、山-欧F1×山葡萄、山-欧F1×欧亚种、山-欧F1×山-欧F14种杂交模式进行种间杂交选育抗寒酿造葡萄品种方面取得的研究进展。并重点讨论了21世纪我国酿酒葡萄抗寒育种发展的技术路线。     

Ampelographic and genetic characterisation of ancestral grapevine (Vitis vinifera L.) accessions present in the Umbria Region (Central Italy)

Summary

During an ongoing effort to recover and preserve local germplasm, 14 accessions of indigenous minor grapevine (Vitis vinifera L.) cultivars from the Umbria Region, Central Italy, were chosen because they had been neglected and were threatened with extinction. Their phenotypic and genetic characteristics were evaluated through an ampelographic study of their shoots, mature leaves, bunches, and berries and by genomic analysis using an international set of nine microsatellite (simple sequence repeat; SSR) markers (VVS2, VVMD5, VVMD7, VVMD27, VrZAG62, VrZAG79, VVMD25, VVMD28, and VVMD32). Comparisons of the SSR profiles of all 14 accessions with grapevine accessions in several databases permitted the identification of unique genotypes, as well as possible synonyms. Information on these older, neglected cultivars will help to reduce the genetic erosion of grapevine germplasm, improve conservation and possible recovery, and assist in the future production of new, distinctive wines.     

利用RAPD标记研究部分葡萄品种的亲缘关系

对47个葡萄品种进行RAPD分析。从100条随机引物中筛选出30个多态性引物用于DNA扩增,共扩出216条DNA带,121条具有多态性,多态性比例为56.01%。通过聚类分析结果表明,供试品种分成5大类,巨峰、红瑞宝、龙宝等巨峰系品种聚为一类。其中龙宝和红瑞宝优先聚在一起。表明葡萄品种间的遗传多样性有明显差异,即使父母本相同的品种,利用RAPD技术也可以分开。聚类图显示,品种间遗传一致度与品种的遗传背景有关,如高妻与红井川,红瑞宝与龙宝优先聚在一起;有的品种虽属同一杂交组合却未能表现最大遗传一致度,如京玉与奥古斯特,在聚类图中处于相邻的位置,说明无性繁殖的葡萄在双亲基因型杂合条件下,有性杂交通过基因重组,杂种后代也可呈现出有较大遗传差异。     

Effect of bearing position on phenolics profiles in the skins of four cultivars of grapevine (Vitis vinifera L.)

Summary

The fan-shaped trellis training system, resulting in grapevines (Vitis vinifera L.) with more than two trunks, has been widely used in northern China. The fruiting zone of grapevines trained to the fan-shaped trellis system is distributed from the top to the bottom of the canopy. The phenolics profiles of the skins of ‘Cabernet Sauvignon’, ‘Merlot’, ‘Chardonnay’, and ‘Italian Riesling’ grape berries were analysed to measure the effect of differences in bearing position on the phenolic compound and anthocyanin compositions of grape berries.The results showed that the upper bearing position significantly increased the concentrations of most phenolic compounds in grape berry skins. Among the non-anthocyanin and anthocyanin phenolic compounds identified in this study, the upper bearing position promoted the accumulation of delphinidin-, cyanidin-, and petunidin-glucosides in the skin of the two red grape cultivars. However, lowering the bearing position was associated with greater methylation of anthocyanins, and the flow of photosynthate towards the biosynthesis of kaempferol. These results were correlated with micrometeorological alterations in the canopy at the different bearing positions. The findings of this study suggest that higher bearing positions can increase the accumulation of phenolic compounds in grape berry skins in four different grapevine cultivars.     

抗病抗寒无核葡萄胚挽救研究

【目的】为了培育抗病抗寒无核葡萄新品种,【方法】以2个种子败育型无核品种‘波尔莱特’、‘红无籽露’作母本,以抗病抗寒的中国野生山葡萄株系‘黑龙江实生’、‘双优’及欧山杂种‘北醇’、‘00-1-10’(‘玫瑰香’ב黑龙江实生’)分别作父本杂交,授粉51 d后将胚珠分别接种于ER和MM4培养基上进行胚挽救,培养60 d后在WPM+BA 0.2 mg.L-1培养基上诱导成苗。【结果】结果表明,2个母本品种胚挽救的适宜培养基不同,‘波尔莱特’作母本适合于ER培养基,‘红无籽露’作母本适合于MM4培养基。2个母本基因型对胚珠的发育率和成苗率的影响差异不大,‘波尔莱特’略优于‘红无籽露’;但4个父本基因型对胚挽救效果影响较大,以欧山杂种‘00-1-10’和‘北醇’作父本的杂交组合胚株的发育率和成苗率明显高于山葡萄‘黑龙江实生’、‘双优’作父本的杂交组合。共获得无核葡萄胚挽救新种质50个株系。【结论】在胚挽救过程中,不同基因型的无核葡萄适宜于不同的基本培养基,欧山杂种比山葡萄更适宜于作杂交的父本。     

UV-C辐射对盆栽葡萄北丰叶片和邻近果穗果实白藜芦醇及其糖苷质量分数的影响

以1a生北丰(Vitis thunbergii×V.vinifera)葡萄盆栽结果树为试材,在果实始熟期采用紫外线C(Ultraviolet C,UV-C)辐射结合韧皮部环剥的方法,研究了UV-C辐射对葡萄叶片和邻近果穗果实白藜芦醇(Resveratrol,Res)及其糖苷质量分数的影响。结果表明,UV-C辐射诱导葡萄果实Res合成积累的敏感性低于叶片,辐射葡萄叶片对邻近果穗上果皮Res的影响远远小于UV-C直接辐射葡萄果穗,且辐射诱导葡萄果皮中Res质量分数与叶片中Res质量分数存在着一定的相关性。当叶片位于果穗下方时,叶片中反式白藜芦醇(trans-Resveratrol,trans-Res)、反式白藜芦醇苷(trans-Piceid,trans-PD)和顺式白藜芦醇苷(cis-Piceid,cis-PD)质量分数表现出环剥处理显著高于不环剥处理,然而果皮中的结果正好相反。当叶片位于果穗上方时,果皮中trans-Res、trans-PD和cis-PD质量分数表现出不环剥处理显著高于环剥处理,暗示葡萄Res可能存在韧皮部运输途径,且运输方向是双向的。此外,UV-C直接辐射葡萄果穗处理果皮trans-Res和cis-PD质量分数显著高于对照和UV-C辐射葡萄叶片处理。种子中仅检测到trans-Res,且仅环剥处理和辐射下方叶片不环剥处理显著高于对照。     

Stilbene Levels in the Tissue and Juice of Muscadine Grapes (Vitis rotundifolia Michx.)

Abstract

Stilbenes are secondary metabolites of a class of non-flavanoid phenolics found in certain species of Vitis. This class of compounds includes resveratrol and a resveratrol glucoside called piceid that exists in trans and cis forms. This study was undertaken to determine the cis and trans forms of both resveratrol and piceid in the skin, pulp, seeds, and juice of nine cultivars of muscadine grapes and three cultivars of Vitis labruscana. Juice samples of the muscadine cultivars ‘Fry’, ‘Hunt’, ‘Magnolia’, ‘Watergate’, ‘Carlos’, ‘Noble’, and ‘Sweet Jenny’, and the V. labruscana cultivars ‘Albermarle’, ‘Miss Blanc’, ‘MidSouth’ and ‘Miss Blue’ were harvested at the full ripe stage and divided into two sub-samples. One subsample was used to extract juice and the other was divided into skins, pulp, and seeds. Sample analysis was performed using HPLC with a UV detector. Sample chromatagrams were compared with those of known standards for quantification. ‘Carlos’ and ‘Magnolia’ had the greatest total stilbene concentration in skin tissue and had bronze skin containing higher concentrations of total stilbenes than any of the black-skinned cultivars tested. Only one bronze skin cultivar, ‘Sweet Jenny’, had lower stilbenes than the dark skin cultivars. Piceid, cis piceid and resveratrol were found in the skins of all cultivars. ‘Carlos’ ‘Magnolia’, ‘Fry’ and ‘Albermarle’ muscadines contained cis resveratrol in the skins of the fruit. ‘Watergate’ was the only cultivar in which resveratrol was detected in seed tissue. ‘Sweet Jenny’ had the highest levels of cis piceid in seed tissue. ‘Albermarle’, ‘Carlos’ and ‘Sweet Jenny’ had significantly greater levels of cis picied in pulp tissue than all other cultivars.     

Physiological response of grapevine cultivars and a rootstock to infection with Phaeoacremonium and Phaeomoniella isolates: an in vitro approach using plants and calluses

In vitro plants and callus culture of two Vitis vinifera cultivars (cv. Baga and Maria Gomes) and one rootstock (R3309, i.e. Vitis riparia var tomentosa × Vitis rupestris) were inoculated with conidia of Phaeoacremonium angustius and Phaeomoniella chlamydospora. Response to infection was determined in plants grown in vitro by assaying growth rates, malondialdehyde (MDA) production (lipid peroxidation) and chlorophyll content and fluorescence. Growth rate and malondialdehyde production were also used to determine resistance of calluses to infection. Infection reduced growth and increased MDA in infected plants and calluses, and reduced chlorophyll content and fluorescence in infected leaves. Symptoms were more evident in plants infected with P. angustius, showing that this species is more virulent to plants and calluses than Ph. chlamydospora. Differences in virulence among strains of Ph. chlamydospora were also found, as 1AS and CAP053 were more virulent (induced more severe decreases of growth and chlorophyll fluorescence, together with higher MDA production in both cultivars) then CAP080. Growth of rootstock plants and calluses was less affected by infection than growth of other cultivars. Contrarily to Baga and Maria Gomes, chlorophyll content and fluorescence of rootstock plants were only affected by P. angustius. Also Baga plants and calluses were more resistant than those of Maria Gomes. These data show different degrees of resistance among genotypes. Reduction of callus production by infection supports the idea that fungus infection may reduce cicatrisation by inhibiting callus formation during grafting or wounding; and therefore, contribute to the entrance of opportunist pathogens. Implications of using in vitro cultures to assay host/pathogen relationship and virulence/resistance degrees among the different genotypes of fungus and grapevines are discussed.     

Growth of Vitis vinifera L. and Vitis aestivalis Michx. as Affected by Temperature

Abstract

Four European (Vitis vinifera L.) winegrape cvs., ‘Semillon’, ‘Pinot Noir,’ ‘Chardonnay’, and ‘Cabernet Sauvignon’, and one American (Vitis aestivalis Michx.) winegrape cv. ‘Cynthiana’, were subjected to three temperature regimes in growth chambers set at 20/15°C, 30/ 25°C, or 40/35°C, for 16/8 hr day/night to determine the influence of temperatures on vine growth and development. In general, the best temperature for shoot and root growth 28 days after temperature treatments was 20/15°C for ‘Semillon’, ‘Cabernet Sauvignon’, and ‘Cynthiana’, and 30/25°C for ‘Pinot Noir’ and ‘Chardonnay’. Although 40/35°C reduced number of leaves, shoots, tendrils, and internodes, total leaf area (LA), and total shoot biomass of all the cultivars, the reduction was more pronounced in ‘Cynthiana’ than in the European cultivars. The average reduction in number of leaves at 40/35°C for the European cultivars was 47%, compared with 92% for ‘Cynthiana’. The two types of grapes adapted differently to high temperature. Shoot growth in the European cultivars continued under high temperature, whereas growth ceased in ‘Cynthiana’. Roots of ‘Cynthiana’, however, were less susceptible to the adverse effect of high temperatures than were the shoots. This study shows that the European cultivars were relatively more tolerant to high temperature than the American cultivar and they have a potential for production of wine in the climate of south central Kansas.     

柑橘品种对不同基因型柑橘衰退病毒的抑制及变异的影响

为研究不同柑橘品种对柑橘衰退病毒（Citrus tristeza virus,CTV）不同基因型的抑制效果,将T36、T30、VT和T3等基因型CTV的毒源分别接种于‘赛蒙斯’甜橙、‘邓肯’葡萄柚、‘墨西哥莱檬’和‘强徳勒柚’,运用RT-qPCR技术检测被接种植株中CTV复制水平的差异。结果表明,‘墨西哥莱檬’最适于CTV各基因型的复制。‘邓肯’葡萄柚中VT、T3和T36基因型的复制水平最低。4种柑橘品种对T36基因型CTV都有明显的抑制作用。通过对CTV的CP基因进行分析发现,VT基因型CTV在‘赛蒙斯’甜橙中发生了较大变异。     

Somatic embryogenesis in a broad spectrum of grape genotypes

Somatic embryogenesis is the preferred method for cell-to-plant regeneration of grapevine. In this study, we tested the embryogenic capacity of anther-derived calli from 59 grape genotypes, representing a diverse group of Vitis vinifera and hybrid varieties, and hybrids and accessions of non-vinifera Vitis species. Most genotypes were tested on two types of media: MST1 medium, which contained plant growth regulators (PGRs) 2,4-dichlorophenoxyacetic acid (2,4-D) and thidiazuron (TDZ), and MSE medium, which contained 2,4-D and 6-benzylaminopurine (BAP). Twenty-four of the grape genotypes produced embryogenic callus on one or both of these media, eighteen of which have not been reported to form somatic embryos before. The results also suggested that the various PGR combinations are differentially effective at inducing somatic embryos in various classes of grape genotypes. For example, seven of the eight V. vinifera conv. occidentalis varieties brought forth somatic embryos on MSE medium, and three out of four American Vitis genotypes produced somatic embryos on MST1 medium. We could not observe any apparent association between frequency of callus formation and embryogenic capacity of the anthers.     

Phenolic Compounds,Antioxidant Activity,Fatty Acids and Volatile Profiles of 18 Different Walnut (Juglans regia L.) Cultivars and Genotypes

The study was carried out to determine phenolic compounds, antioxidant activity, total lipids, saturated fatty acids, unsaturated fatty acids, and volatile organic compounds (VOCs) of 18 different walnut (Juglans regia L.) cultivars and genotypes grown in Usak province, Turkey. The biochemical compounds were identified for each cultivar and genotype and a comparative evaluation was carried out. The results showed that most of the biochemical compounds are significantly varying among each other (p?<?0.05). The existence and abundance of the biochemical compounds in the cultivars and genotypes have not only found to be a cause of differentiation, but they have been important parameters for the similarities among the cultivars and genotypes. Total phenolic content and antioxidant activity ranged from 750.67 to 1245.64?mg GAE ml^?1 and from 42.46 to 56.50% in kernels, respectively. The highest contents of the phenolic compounds were noted from gallic acid (11991?mg kg^?1), ellagic acid (1057.16?mg kg^?1), catechin (425.4?mg kg^?1), and rutin (216.6?mg kg^?1), as an average of all cultivars and genotypes. The highest lipid contents were noted from ‘Franquette’ and ‘Fernette’ as 64.28% and 63.26%, respectively, while the lowest content was noted from ‘Oguzlar 77’ (52.52%). A total of 42 VOCs were described from the 18 different walnut cultivars and genotypes. Overall, results showed that the phenolic profiles, fatty acids composition, and VOCs play an important role in similarities and diversities among the cultivars. According to the results, 7 different clusters were developed from the 18 cultivars and genotypes. Herein, ‘Fernor’, ‘Maras 18’, ‘Arslan Local Type’ and ‘Cisco’ cultivars were found to have the lowest biochemical compositions. The superior cultivars or genotypes, in terms of the abundance of the biochemical composition, were found to be ‘Gulizar Hanim Type’, ‘Lara’, ‘Pedro’, ‘Tulane’, ‘Fernette’, ‘Balkan’ and ‘Franquetta’, where they were found to develop 3 clusters by 1:3:3, respectively.

     

葡萄果实发育后期脱落酸来源的研究

  以‘品丽珠’(Vitis vinifera L. cv. Cabernet-Franc) 葡萄为试材, 对发育后期果实中脱落酸(ABA) 的来源进行了研究。结果表明, 叶片合成的ABA 可能是果实中ABA 的重要来源, 而根部ABA 的合成对果实中ABA 含量贡献不大。种子中ABA 的合成可能不是果肉中ABA 的重要来源。果实具有独立合成和降解ABA 的能力。对果实内ABA 含量变化的代谢调控问题进行了讨论。     

Determination of grafting compatibility of grapevine with electrophoretic methods

This study was carried out to determine the efficiency of electrophoretic methods in predicting graft incompatibility of grape cultivars with American rootstocks. Three isoenzyme systems (peroxidase, PER; esterase, EST; acid phosphatase, AcPH) and total protein profiles were obtained in 15 grape cultivars (Vitis vinifera L.) and 12 American rootstocks. Compatibility levels were determined by the band similarities. Field compatibilities were also calculated. Results showed that incompatibility exists between different cultivar–rootstock combinations. AcPH and total protein profiles of the cultivar–rootstock combinations could be suggested to use for forecasting graft incompatibility.     

中国葡萄属野生种抗黑痘病的鉴定研究

本文通过田间自然鉴定、田间接种鉴定和室内离体接种鉴定的方法,研究了起源于我国的葡萄属18个种或变种、88个株系对黑痘病的抗性。结果表明:所有野生种的果实几乎不感黑痘病,叶片表现为高度的或较强的抗性。     

Uneven ripening in ‘Bangalore blue’ grape (Vitis vinifera × Vitis labrusca) berries in India is linked to seed viability

Uneven ripening (UR) is a physiological disorder of unknown origin in ‘Bangalore blue’ grape (Vitis vinifera × Vitis labrusca) leading to wine of inferior quality. A preliminary study found wide variations in total dehydrogenase activity (TDH) of seeds from unevenly ripe berries. In our experiments, gibberellin (GA₃) applied to young grapes increased seed TDH activity and reduced the incidence of uneven ripening to 2% compared with 35% in the control. In contrast, paclobutrazol (PBZ) decreased TDH activity and increased the incidence of the disorder to 58%. GA₃-treated berries had higher concentrations of sucrose and TDH activity in seed representing mature seeds with high viability. In contrast, PBZ-treated and control berries had higher concentrations of glucose and lower TDH activity, indicating immature seeds with low viability. These results suggested that competition among developing berries can lead to differences in seed gibberellin content, seed viability and the rate of berry growth resulting in green, purple, and black berries at harvest. The study established the role of seed viability in uneven ripening and demonstrated that the incidence of the disorder is reduced by the application of GA₃ to immature berries.     

Screening of cold-resistant seedlings of a Chinese wild grape (Vitis piasezkii Maxim var. pagnucii) native to loess plateau of eastern Gansu province,China, as rootstocks

Seedlings of a Chinese wild grape (Vitis piasezkii Maxim var. pagnucii) native to loess plateau of Eastern Gansu province, China, were evaluated to screen cold-resistant rootstocks in Lanzhou area. After 14-year investigation two selections of LDP-191and LDP-294 were screened as rootstocks for two table grape cultivars, ‘Fujiminori’ and ‘Red Globe’, respectively. The two graft unions demonstrated very high cold-resistance as well as good graft compatibility. Furthermore, they could survive through low temperatures in winter without soil coverage together with good fruit quality of the cultivars grafted.     

Origin and genetic diversity of Tunisian grapes as revealed by microsatellite markers

Ten SSR loci, previously developed for grapevine, were analyzed to evaluate the genetic variability, cultivar relatedness, and parentage in a collection of 61 autochthonous Vitis vinifera cultivars from Tunisia.The number of alleles per locus ranged from 6 to 11, while the number of genotype patterns varied between 10 and 21. The expected heterozygosity varied between 0.621 and 0.855 and the observed heterozygosity was higher than 0.9 at 4 loci (VVMD28, VVMD5, VVIP31 and VVS2) indicating that the SSRs were highly informative.Cluster analysis using unweighted pair group method with arithmetic averaging (UPGMA) suggested 14 groups among studied cultivars and 53 grapevine denominations out of 61 were unequivocally distinguished, with all accessions showing at least one-specific combination of alleles.On the other hand, in order to overcome the existing confusion in Tunisian grapevine nomenclature, of the analyzed homonymous pairs of cultivars, only ‘Balta 2’ and ‘Balta 3’ have shown identical allelic profiles, consistent with their being the same genotype. Hence, nomenclature distinction is meaningless and only one denomination should be retained.Due to the high overall power of exclusion (Q) (greater than 99.99%) and to the absence of null alleles, the set of microsatellite loci used is appropriate to determine parentage in Tunisian grapevines beyond any reasonable doubt. The analysis of fingerprints indicated that the Tunisian grape vines have evolved through out crossing between five possible parents: Balta 1, Beldi Baddar, Beldi Rafraf, Beldi Local Rafraf and Khedhiri 3.