Evaluation of conditions for in vitro storage of commercial and minor grapevine (Vitis vinifera L.) cultivars

In vitro culture represents a tool for the ex situ conservation of a high number of sanitised plants in a reduced space. However, the culture media and/or other growing conditions need to be optimised to minimising plant growth and storage cost. Growth on MW medium was evaluated in the commercial cultivars ‘Airén’, ‘Bobal’, ‘Chardonnay’, ‘Garnacha Blanca’, ‘Moscatel de Alejandría’, ‘Moscatel de Grano Menudo’, ‘Pedro Ximénez’, ‘Pinot Blanc’, ‘Pinot Gris’, ‘Sauvignon Blanc’, and ‘Tempranillo’; the minor cultivars ‘Chelva’, ‘Valencí Negre’, ‘Valencí Blanc’, and ‘Verdil’; and the endangered cv. ‘Esclafacherre’. Different growth rates were observed among cultivars: those with faster growth need to be subcultured every 1.5–2.0 months; those with the slowest growth every 3.5–4.0 months. The effect of halving the sucrose in MW reduced the growth of the cultivars that grew faster without compromising survival. When IBA was removed from MW, growth was also reduced in some cultivars. Therefore, small modifications of the MW composition are adequate for grapevine in vitro storage under standard incubation conditions. This is an advantage with respect to the change of temperature used in other work to achieve growth reduction, and allows the use of the same chamber for different in vitro culture procedures.     

A 3-Year Adaptation Study of Three Distinct Grapevine Cultivars under Midwestern Field Conditions

Simultaneous responses of American (Concord), European (Pinot Noir), and American-European hybrid (Traminette) cultivars to seasonal and diurnal variation in field environmental conditions were assessed to propose suitable cultivars for potential use for selection and breeding in grapevines in dry areas. ‘Pinot Noir’ had higher gas exchange parameters, higher water potential, and chlorophyll fluorescence, while ‘Concord’ had the lowest values in all three seasons and experienced moderate stress by the end of each season. The dry conditions in 2005 caused water potential to decline sharply compared to values recorded in 2004 and 2006. ‘Pinot Noir’ and ‘Traminette’ exhibited a mild stress, while ‘Concord’ exhibited a moderate stress in 2006. Photosynthetic rate decreased slightly during the summer with a slight recovery at the beginning of September, and did not differ across the seasons among cultivars. Stomatal conductance followed the same pattern. However, in both 2004 and 2005, stomatal conductance dropped sharply late in the season because of frequently occurring high temperatures that were greater than 32 °C. Maximum morning values of Fv/Fm were within the optimal range. The study suggests the presence of marginal photoinhibition during mid-summer in Concord cultivars, because no full recovery of photochemical efficiency early in the evening was observed. The European cultivar (Pinot Noir) and the American-European hybrid (Traminette) are more adapted to field conditions in Kansas than Concord cultivars. However, high cold-mortality of Pinot Noir due to high sensitivity to cold winter temperature makes the hybrid cultivars the most suitable cultivar for Midwest regions with relatively cold winters.     

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.     

不同酿酒葡萄品种白藜芦醇合酶基因的克隆和序列分析

以红葡萄品种“赤霞珠”、“品丽珠”、“黑比诺”和白葡萄品种“雷司令”为试材,采用改良CTAB法提取葡萄叶片组织中的DNA,并扩增得到4个酿酒葡萄品种的白藜芦醇合酶基因部分片段.结果表明:红葡萄品种“赤霞珠”、“品丽珠”、“黑比诺”的扩增产物均为1 419 bp,白葡萄品种“雷司令”扩增产物为1 417 bp.通过序列多重比较分析,所得到的4个白藜芦醇基因片段与Genbank中河岸葡萄(AF128861)的该基因序列同源性较高(90％以上),外显子区同源性高于内含子;红色葡萄姊妹品种之间、红色葡萄不同品种之间、红色葡萄与白色葡萄品种之间的序列同源性依次递减.     

几个酿酒葡萄品种在广夏第三种植基地的栽培表现

以15个酿酒葡萄品种为试材,采取田间试验和室内检测相结合的方法,对广夏第3种植基地品种园的15个酿酒葡萄品种的主要物候期、生长结果习性及果实主要性状等进行了调查。结果表明:"黑比诺"抗寒、稳产、品质好,作为早熟的红色品种,具有推广价值;"赛美容"果穗大,丰产,品质优,可适当扩大栽培;"灰比诺"酸度低,风格较独特,可选择栽培。     

贺兰山东麓六个酿酒葡萄品种抗寒性比较

以宁夏贺兰山东麓主栽的"赤霞珠""美乐""蛇龙珠""马瑟兰""西拉""霞多丽"6种酿酒葡萄发育良好的1年生枝条为试材,在4℃的对照及-18、-20、-22、-24℃的低温条件下模拟低温冻害,通过测定其电导率值、脯氨酸(Pro)含量、丙二醛(MDA)含量3项生理指标,以期鉴定贺兰山东麓大量种植的欧亚良种酿酒葡萄的抗寒性,提高贺兰山东麓葡萄产区的越冬冻害防御能力。结果表明:随着处理温度不断下降,6种酿酒葡萄品种中的枝条电解质渗出率增大;脯氨酸含量、丙二醛含量显著增加。6个试验品种的抗寒性强弱为"赤霞珠">"霞多丽">"蛇龙珠">"马瑟兰">"美乐">"西拉"。     

宁夏贺兰山东麓不同葡萄品种对霜霉病的抗性鉴定

为了给葡萄抗性品种的选育和葡萄霜霉病的防治提供科学依据,对16个不同酿酒葡萄品种和19个鲜食葡萄品种的抗病性进行了抗性测定。结果表明,室内回接与田间调查结果一致,16个酿酒葡萄品种可划分为4类;19个鲜食葡萄品种可划分为4类;从发病时间来看,抗性与发病时间存在一定相关性,抗性越强发病越晚。随即将测定结果采用DPS软件进行聚类分析,在酿酒葡萄抗性反应谱系图上,第1类为抗性稍强品种,为白比诺、灰比诺和赛美蓉;第2类为抗性中等品种,由白长相思和神索等8个品种组成;第3类是稍感病品种,包括赤霞珠等4个品种;第4类是感病品种,只有品丽珠1个。在鲜食葡萄谱系图上,第1类为六月紫等7个抗性稍强品种;第2类是新华1号和巨峰;第3类是稍感品种,由奥古斯特等6个品种组成;第4类为力扎马特等4个感病品种。     

葡萄果实中（–）–α–萜品醇的积累与其合成酶基因Vvter表达的关系

 以不同发育期的‘赤霞珠’和‘雷司令’葡萄果实为试材,应用气相色谱质谱联用技术和荧光实时定量PCR技术,研究了果实发育过程中游离态和糖苷态（–）–α–萜品醇含量与（–）–α–萜品醇合成酶[Vitis vinifera（-）-α-terpineol synthase]基因Vvter表达的关系。结果表明,糖苷态（–）–α–萜品醇在两个品种果实发育过程中均没有明显变化,游离态（–）–α–萜品醇在‘赤霞珠’中表现为随果实发育而降低,而在‘雷司令’中呈现先下降再上升后下降的趋势,同一品种中（–）–α–萜品醇总含量的变化与Vvter基因的表达成正相关。     

A comparison of the response of different citrus fruit cultivars to storage temperature

A comparison was made of the response to storage temperatures (0–17°C) of grapefruit ‘Shamouti’ and ‘Valencia’ oranges, and lemon fruit. There was no chilling injury (CI) and the incidence of rot was relatively low at 12°C or higher for all cultivars. At low storage temperatures, however, pitting and subsequent rot development in grapefruit and in ‘Shamouti’ and ‘Valencia’ oranges were highest at 6°C, while in lemon the incidence of these disorders rose with the decrease in storage temperature, and was highest at 2°C. Grapefruit and ‘Shamouti’ oranges exhibited the highest susceptibility to CI, lemon showed an intermediate susceptibility, and ‘Valencia’ orange a very low one. In grapefruit, severe peel pittings that developed in cold storage were covered by mold rots during cold storage and shelf-life, but in ‘Valencia’ and particularly in ‘Shamouti’ oranges the incidence of decay during cold storage was low. During shelf-life, a pronounced CI-induced increase in the incidence of mold rots was evident. This breakdown during shelf-life may be associated with microscopic peel injuries which are formed during cold storage and covered by mold rots during shelf-life. Increased susceptibility of the cultivar to CI could be correlated with an increase in ethanol content at low temperatures.     

毛欧杂种葡萄‘NW196’花色苷组成与相关基因表达分析

以不同发育期毛欧杂种葡萄‘NW196’为试验材料,利用HPLC–MS检测其果皮中花色苷的组成及含量,Real-time PCR检测花色苷合成相关基因表达水平,并与欧亚种‘赤霞珠’比较。结果表明,成熟后期‘NW196’果皮中花色苷总量显著高于‘赤霞珠’;采收期‘NW196’葡萄果皮中共检测到23种花色苷,成熟后期单糖苷所占比例高于双糖苷,其中含量最高的是花翠素–3–O–葡萄糖苷,3’5’–羟基取代花色苷比例高于3’–羟基取代花色苷;与欧亚种‘赤霞珠’相比,‘NW196’果皮中酰化和甲基化花色苷比例较低。结论:毛欧杂种‘NW196’花色苷组成与欧亚种‘赤霞珠’差异明显,这与其品种特性有关,并受到类黄酮代谢路径相关基因的调控;此外,花色素双糖苷主要在转色初期合成。     

Effect of temperature on pollen germination and pollen tube growth of four cultivars of mango (Mangifera indica L.)

Summary

The effect of a constant (10, 15, 20 or 25°C) or a diurnal maximum/minimum (15/5, 20/10, 25/15 or 30/20°C) incubation temperature on in vitro pollen germination and pollen tube growth in the pistils of two poly-embryonic (‘Kensington’ and ‘Nam Dok Mai’), and two mono-embryonic (‘Irwin’ and ‘Sensation’) mango cultivars was studied. In in vitro experiments where pollen was incubated in a liquid germination medium for 24 h in darkness, little difference was found between pollen germination of mono- and poly-embryonic cultivars. Averaged over the four cultivars, 53.9% of pollen germinated at 10°C, this increased to 76.2% when the incubation temperature was increased to 15°C, thereafter up to 25°C the percentage germination remained stable but germination decreased slightly to 68.2% at 30°C. Similarly, there was no difference in percent germination between cultivars when pollen was incubated under diurnal temperature regimes. Mean pollen germination of all four cultivars was 52.3% at 15/5°C and pollen germination increased by 10% when the temperature was raised to 30/20°C. When self-pollinated flowers were incubated for 24 h on a semi-solid agar medium at 10°C, pollen tube growth of the four cultivars was retarded and no pollen tubes reached the ovaries. As the temperature was increased from 15 to 25°C, the mean number of pollen tubes in ovaries increased from 0.04 to 0.25. At 30°C, the mean number of pollen tubes that entered ovaries decreased to 0.04. After incubation under diurnal temperature regimes, the mean number of pollen tubes in ovaries of all four cultivars at 15/5°C was 0.23 and increased to 0.42 when the temperature increased to 30/20°C. At each incubation temperature, there were significant differences in pollen tube growth between cultivars, but there were no differences between the temperature response of pollen from mono- and poly-embryonic cultivars.     

Seed germination of seven pepper cultivars at constant or alternating high temperatures

Summary

Seeds of seven pepper (Capsicum annuum L.) cultivars (Anaheim TMR 23, California Wonder 300, Coronado, Jalapeno M, Ma Belle, Mercury, and Yolo Wonder B) were germinated at constant day and night temperatures of 25,30,35 and 40°C or at alternating temperatures of 40/25,40/30 and 40/35°C for 14 days. Germination percentages and rates were similar at 25 and 30°C. Largest differences in cultivar responses occurred at 35°C where germination percentages ranged from 24 to 96%, and rates, calculated as summation of the number of seeds germinated on a given day divided by day number, varied from 3 to 26 (theoretical maximum value of 100). At 40°C, germination percentages were less than 5% and rates were less than one for all cultivars. Cultivars with the most heat tolerance were ‘Mercury’ and ‘Yolo Wonder B’. At alternating temperatures, germination percentages and rates were higher than those at constant 40°C. The increases were greatest when the temperature was lowered by 15°C (40/25°C) and least when temperatures were lowered by 5°C (40/35°C). Tetrazolium tests showed that a large percentage of the ungerminated seed was still viable from the highest temperature. At lower temperatures, fewer ungerminated seeds were viable with no viable ungerminated seeds from the lowest temperature.     

The effects of daminozide and under-soil heating on the growth of radish under winter conditions

The growth of radish cultivars ‘Saxa’, ‘Saxerre’, ‘Serra’, ‘Istar’, ‘Cherrybelle’ and ‘Robino’ was compared at 7 and 12°C under winter glasshouse conditions. ‘Serra’ alone produced marketable storage roots by 63 days at 7°C, but root weight was no greater at 12 than at 7°C; only ‘Robino’ produced significantly larger roots at the higher temperature. Under-soil heating applied to ‘Cherrybelle’ and ‘Robino’ grown at an air temperature of 7°C increased root growth, particularly in ‘Cherrybelle’. Daminozide treatment at 7°C increased total and storage root weight and root:shoot ratio of both cultivars. At 12°C, the only significant effect of daminozide treatment was reduced shoot weight in ‘Robino’.     

High-frequency somatic embryo production from unfertilized ovules of grapes

Somatic embryos were produced in large numbers from cultured, unfertilized ovules of Vitis vinifera, cultivars ‘Cabernet Sauvignon’ and ‘Grenache’, and of the hybrid grape ‘Gloryvine’ (V. vinifera × V. rupestris). Callus of nucellar origin was produced by culturing the ovule explants initially in Nitsch medium containing 5 μM 2,4-dichlorophenoxy acetic acid (2,4-D) or 5 μM β-naphthoxyacetic acid (NOA) plus 1 μM benzyladenine (BA), and then in a medium supplemented with NOA (10 μM) plus BA (1 μM). Embryos were produced when callus was transferred to basal medium containing no auxin or cytokinin. Secondary and tertiary embryos were formed by budding of primary somatic embryos. Prolonged culture of ovular callus in NOA (10 μM) plus BA (1 μM) gave rise to cell aggregates which developed into free-floating roots when transferred to basal medium.     

中国野生葡萄抗霜霉病相关基因VpPR4b及其启动子的克隆和功能分析

以中国野生葡萄(Vitis piasezkii)‘留坝-8’(抗霜霉病)和欧亚种葡萄(V.vinifera)‘黑比诺’(易感霜霉病)为材料,克隆得到抗霜霉病相关基因VpPR4b和VvPR4b。序列分析发现VpPR4b和VvPR4b核苷酸序列相似性为98.61%。VpPR4b编码区为432 bp,编码143个氨基酸,含有BARWIN保守结构域,属于典型的Ⅱ型PR4蛋白。VpPR4b和VvPR4b均可在霜霉菌侵染后诱导表达。亚细胞定位表明VpPR4b定位于细胞膜上。分别从‘留坝-8’和‘黑比诺’中克隆得到了VpPR4b和VvPR4b的启动子,其序列相似性高达95.65%,并且含有相似的顺式作用元件。通过酵母单杂交验证了转录因子WRKY40和WRKY75可以结合VpPR4b启动子。本研究的结果表明PR4b可能在葡萄抵御霜霉病侵染过程中发挥一定作用。     

Temperature effects on dormancy,bud break and spear growth in asparagus (Asparagus officinalis L.)

Summary

The effects of the length of chilling, chilling temperature and growing temperature on dormancy of asparagus crown buds and subsequent rates of spear growth were examined. The results showed that prior chilling enhanced bud break at low growing temperatures and stimulated the growth of spears.Thus, chilling should facilitate commercial production by hastening bud break and spear growth rates at lower temperatures. If sufficient chilling was given, the minimum temperature for rapid bud break was approx. 12.5°C for ‘Rutgers Beacon’ and ‘Jersey Giant’, and around 10°C for ‘UC 157’ and ‘Apollo’. The optimum chilling temperature appeared to be closer to 5°C than to 10°C or 2°C for ‘Rutgers Beacon’ plants grown at 12.5°C. Increasing the growing temperature had a significant effect on the relative spear growth rate (RSGR) in all cultivars. Prior chilling had no effect on the RSGR for ‘Dariana’ and ‘Apollo’; but, for ‘UC 157’, chilling plants at 5°C for 5 or 10 weeks increased growth rates at 12.5°C and at 20°C. These results demonstrate that release of bud dormancy and spear growth rates depended not only on the growing temperature, but also, at least in some cultivars at some temperatures, on the duration and temperature of chilling during the previous Winter.     

Effect of temperature on growth and flowering of litchi (Litchi chinensis Sonn.) cultivars

Summary

Moderate day/night temperatures (20/15° v. 15/10°C) increased vegetative growth and reduced flowering in the seven litchi cvs Tai So, Bengal, Souey Tung, Kwai May Pink, Kwai May Red, Salathiel and Wai Chee. At higher temperatures (25/20° and 30/25°C), vegetative growth was promoted further and flowering eliminated. Temperature also influenced the type of inflorescence formed. More leaves were formed on the panicles of trees growing at 20/15° than at 15/10°C. All terminal shoots on all cultivars produced panicles at 15/10°C. The relative order for the amount of flowering at 20/15°C was: ‘Wai Chee’>‘Salathiel’>‘Kwai May Pink’>‘Tai So’>‘Bengal’>‘Souey Tung’>‘Kwai May Red’. Cultivars which were vigorous at high temperatures produced fewer panicles at 20/15°C and fewer leafless panicles at 15/10°C. Only small differences were observed in the leaf water potential and the nutrient status of the shoots at different temperatures. Vigour and flowering of the cultivars in the glasshouse generally reflected field performance in subtropical Australia (Lat. 27°S). Low vigour could be useful for selecting litchi cultivars for good fruiting in environments with warm autumns and winters.     

‘粉红亚都蜜’葡萄NAC转录因子基因VvDRL1的功能初步分析

以欧洲葡萄‘粉红亚都蜜’为材料,利用同源克隆法获得1个NAC转录因子,命名为VvDRL1（GenBank：XP-002281816）。该基因全长840 bp,编码280个氨基酸,与‘黑比诺’葡萄中该基因的同源性为100%,但是基因组DNA序列中存在6处单核苷酸突变。瞬时转化洋葱表皮细胞进行亚细胞定位分析,VvDRL1基因主要在细胞核内表达,属于核蛋白;通过农杆菌介导的叶盘法获得稳定整合的转基因烟草,T2代转基因植株生长迟缓,株高、叶面积和叶片细胞面积约为野生型烟草的60%、34%和31%,转基因植株叶片出现向内卷曲现象,利用石蜡切片进行显微结构观察,转基因植株主叶脉上表皮下缺失2 ~ 3层厚壁细胞,且海绵组织内的空隙明显多于野生型植株。研究结果表明,VvDRL1在调控植株的形态发育方面起着重要作用。     

Long-day control of flowering in everbearing strawberries

Summary

Photoperiod and temperature control of flowering in a number of perpetual-flowering or everbearing strawberry cultivars of widely varying pedigree has been studied in controlled environments. Flower bud initiation in the cultivars ‘Flamenco’, ‘Ridder’, ‘Rita’ and ‘Rondo’ was significantly advanced by long-day (LD) conditions at temperatures of 15°C and 21ºC; while, at 27ºC, flowering took place under LD conditions only. Some plants of the seed-propagated F₁-hybrid ‘Elan’, raised at 21°C, also flowered under short-day (SD) conditions at 27°C, but reverted to the vegetative state after a few weeks when maintained under these conditions. When vegetative plants growing in SD at 27°C were transferred to LD conditions at the same temperature, they consistently initiated flower buds and started flowering after about 4 weeks. At such a high temperature, flowering could thus be turned on and off by switching between SD and LD conditions. This applied to all the cultivars studied. Also the cultivar ‘Everest’, which was tested only at 21°C, produced similar results. Night interruption for 2 h was effective in bringing about the LD response. At 9°C, flowering was substantially delayed, especially in ‘Flamenco’ and, at this temperature, flowering was unaffected by photoperiod. Runner formation was generally promoted by high temperature and SD conditions, but the photoperiodic effect varied between experiments. We conclude that everbearing strawberry cultivars, in general, whether of the older European-type or the modern Californian-type originating from crosses with selections of Fragaria virginiana ssp. glauca, are qualitative (obligatory) LD plants at high temperature (27°C), and quantitative LD plants at intermediate temperatures. Only at temperatures below 10°C are these cultivars day-neutral.     

硒对水分胁迫下葡萄幼苗光合特性的影响

以酿酒葡萄赤霞珠扦插盆栽苗为试材,研究硒对水分胁迫下葡萄幼苗光合特性的影响,以期为硒在葡萄抗旱栽培方面的应用提供参考依据.结果表明:随水分胁迫时间的延长,葡萄光合作用逐渐下降.水分胁迫条件下,葡萄叶面喷施一定浓度(0.5～1.5 mg/L)的亚硒酸钠均能提高叶绿素含量,但以0.5 mg/L的亚硒酸钠溶液处理效果最好,可以缓解水分胁迫对葡萄光合作用的影响;当葡萄叶面喷施低浓度(0.5 mg/L)的亚硒酸钠时,能缓解净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)及重度水分胁迫以前胞间CO2浓度(Ci)的下降,同时还能促进重度水分胁迫以后胞间CO2浓度(Ci)的升高;当喷施高浓度(1.0～1.5 mg/L)的亚硒酸钠时,会促进这些光合相关指标的降低.