油菜素内酯和脱落酸调控葡萄果实花色苷合成的研究

【目的】为研究2,4-表油菜素内酯(2,4-epibrassinolide,EBR)和脱落酸(abscisic acid,ABA)处理对葡萄花色苷合成与可溶性固形物含量、果皮苯丙氨酸解氨酶(PAL)和类黄酮糖基转移酶(UFGT)活性以及果实内源ABA含量的影响,探索油菜素内酯调控葡萄果实成熟及花色苷合成的机理。【方法】以酿酒葡萄‘赤霞珠’(Cabernet Sauvignon)和‘烟73’(Yan 73)为试材,在葡萄转色前分别用0.1、0.4、0.8 mg.L-1EBR,1mg.L-1Brz(brassinazole,BR生物合成抑制剂)和200 mg.L-1ABA,均匀喷施于葡萄果实,在葡萄成熟过程中测定葡萄果皮花色苷含量及PAL和UFGT酶活性,同时测定果实ABA和可溶性固形物含量。【结果】在果实着色初期,‘赤霞珠’和‘烟73’葡萄果皮PAL和UFGT活性及果实内源ABA含量均逐渐升高,当果实接近成熟花色苷含量基本稳定时ABA含量开始降低。与对照相比,0.4mg.L-1EBR和200 mg.L-1ABA处理显著增加了果实内源ABA含量,提高了果皮PAL和UFGT活性,促进了果皮花色苷的合成和果实可溶性固形物的积累。0.1 mg.L-1和0.8 mg.L-1EBR处理总体增加了果实内源ABA含量,促进了花色苷的合成和可溶性固形物的积累,并提高了UFGT和PAL酶活性,但比0.4 mg.L-1EBR处理提高的幅度小,且差异显著。1 mg.L-1Brz处理使果实ABA的合成推迟,果实可溶性固形物含量、果皮PAL和UFGT活性以及花色苷含量均低于对照,但差异不显著。【结论】外源EBR和ABA处理促进了葡萄成熟和花色苷合成,在不同浓度EBR处理中,以0.4 mg.L-1处理效果较好;内源ABA可能参与了EBR对葡萄成熟和花色苷合成的调控。     

Abscisic acid and 2,4-dichlorophenoxyacetic acid affect the expression of anthocyanin biosynthetic pathway genes in ‘Kyoho’ grape berries

Summary

The effects of abscisic acid (ABA) and 2,4-dichlorophenoxyacetic acid (2,4-D) on the expression of seven anthocyanin biosynthetic pathway genes in ‘Kyoho’ grape berries were investigated. In untreated berries, the expression of the UDP-glucose-flavonoid: 3-O-glucosyltransferase (UFGT) gene was detected only at 42 d after full bloom (DAB), whereas the phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone-3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR) and leucoanthocyanidin dioxygenase (LDOX) genes were expressed throughout the growing period. ABA increased anthocyanin content in the skin and the expression of PAL, CHS, CHI, DFR and UFGT genes at 7 d after treatment. In contrast, 2,4-D inhibited the accumulation of anthocyanin and the expression of all the genes examined. The results clearly show that the anthocyanin levels resulting from the application of ABA and 2,4-D were correlated with the expression of anthocyanin biosynthetic pathway genes.     

Cell walls of loosening skin in post-veraison grape berries lose structural polysaccharides and calcium while accumulate structural proteins

Changes of mechanical properties, ultrastructure, cell wall components and wall-bound peroxidase in grape skin were studied prior to and after veraison in cv. Golden Muscat (Vitis vinifera × V. labrusca). Skin loosened continuously after veraison. Ultrastructural changes of the loosening skin included wall swelling in the epidermis and subepidermis cells, degradation of the middle lamella in the inner cells, and wall surfaces becoming “wavy”. Apoplast acidification took place in post-veraison berries. Contents of wall structural components, e.g. insoluble pectin, hemicellulose, cellulose and wall-bound calcium in the skin decreased, while both content of SDS-insoluble amino acids (especially hydroxyproline) and the activity of wall-bound peroxidase increased significantly after veraison. The activity of the enzyme was found localized mainly in the walls of the outermost four layers of cells including the epidermis and subepidermis tissues. The results indicated that, during loosening of skin, the cell walls in the skin constantly lose structural polysaccharides and calcium. Meanwhile, structural protein incorporation and phenolic cross-linkage formation became active especially in the walls of epidermis and subepidermis cells.     

Temperature effects on corm dormancy and growth of Zephyra elegans D.Don

Experiments were performed with the Chilean geophyte Zephyra elegans, a potential cut flower, to evaluate the effect of corm weight and storage temperature on corm dormancy, and to determine the effect of day and night growing temperatures on its growth and flowering. Z. elegans has a deciduous and synanthous growth habit and the corm is replaced annually. Dormant corms were stored at different constant temperatures or temperature combinations from 20 to 40 °C. Corms released from their dormancy were grown at 15/10, 20/15, or 25/20 °C day/night temperatures. Corms of various weights were planted at the same date after being stored dry at 25 °C for 22 weeks. They all emerged 19–38 days after planting, showing that dormancy release was not affected by corm weight. A 20-week corm storage treatment at a constant 25 °C resulted in the most rapid corm sprouting. Sprouting percentage was reduced at higher or lower storage temperatures. Temperature also affected plant growth. When plants were grown at 15/10 or 20/15 °C they emerged and flowered more rapidly than when they were grown at 25/20 °C. The latter growing temperature also resulted in poor flower quality.     

Day and night temperatures,daily light integral,and CO2 enrichment affect growth and flower development of Campanula carpatica ‘Blue Clips’

Campanula carpatica Jacq. ‘Blue Clips’ plants were grown in a greenhouse under nine combinations of day and night temperatures created by moving plants every 12 h among three day/night temperatures (15, 20, and 25°C). At each temperature, there were three daily light integrals (DLI; 4.2, 10.8, and 15.8 mol m⁻² per day, averaged over the experimental period) created with varying supplemental light, and ambient (≈400 μmol mol⁻¹) and enriched (≈600 μmol mol⁻¹) CO₂ concentrations. Time to flower was closely related to average daily temperature (ADT), and was not significantly affected by the day or night temperatures delivered to achieve a specific ADT. Time to flower was not largely affected by DLI or CO₂ enrichment. As plant ADT increased between 15 and 25°C, flower diameter decreased about 1 mm per degree and was not related to the difference between day and night temperatures (DIF). Flower diameter was smallest and least sensitive to changes in temperature at lower DLI and at ambient CO₂ levels. There were 10 less flower buds and 0.3 g less dry mass per plant at first flower for every 1° increase in plant ADT at high and medium DLIs. Flower bud number and dry mass were relatively low and less sensitive to changes in ADT at low DLI, and increased slightly with CO₂ enrichment at medium and high but not at low DLI. Plant height was not related to ADT, but increased linearly as DIF increased from −6 to 12°C at all DLIs, but the response was stronger under low DLI than high and medium DLIs. Flower bud number and dry mass were correlated closely with the ratio of DLI to daily thermal time (base temperature of 0°C). Flower bud number and dry mass were highest when C. carpatica plants were grown at 15°C with a DLI of 10–15 mol m⁻² per day.     

金雀异黄素和环鸟苷酸调控离体葡萄果实花青苷积累

以离体‘巨峰’葡萄为材料,观察到金雀异黄素（GNT）对果皮花青苷积累的促进效应,而且证明GNT的诱导过程（约10 ~ 12 h）不需要光照,但其后的花青苷积累却依赖于光照。如果在GNT预处理后6 h内用环鸟苷酸（cGMP）处理,可以明显抑制GNT的促进效应,但在GNT预处理后12 h再用cGMP处理,则抑制效应消失。半定量RT-PCR检测表明,光和GNT诱导葡萄果皮花青苷合成相关基因PAL、CHS和UFGT表达量上调,而对LDOX表达量无明显影响;cGMP处理抑制了GNT诱导的PAL和CHS等基因表达的促进作用,说明GNT和cGMP相互拮抗,共同调控葡萄果皮花青苷积累。     

Temperature and irradiance effects on Sandersonia aurantiaca flower shape and pedicel length

Sandersonia flowers can vary from being lantern- to tubular-shaped. Lantern-shaped flowers are considered to be most commercially desirable when the ratio of the widest to the narrowest diameters of a flower, termed the ratio of hips to waist, is 1.5 or greater. Pedicel length can also affect the overall appearance of a flower stem. Short pedicels are considered more desirable. The influences of temperature and irradiance on Sandersonia aurantiaca flower shape and pedicel length were examined in controlled environment (CE) growth rooms. The growing environments were at constant temperatures of 15, 18, 21, 24 or 27 °C; or day/night temperature differentials of −6, 0, +6, +12, or +18 °C, with a mean of 21 °C. Photosynthetic photon flux densities (PPFD) were 700, 460 or 210 μmol m⁻² s⁻¹. At harvest, measurements were made of the widest and narrowest diameters of the oldest flower on each stem and the ratio of these parts was calculated. The pedicel length of the oldest flower was also measured. Environments with constant temperatures below 23 °C and the least day/night temperature differentials (−6, 0 and +6 °C) produced flowers of the most desirable shape, defined as having a hips to waist ratio of 1.5 or greater. At these temperatures, irradiance had a variable effect on flower shape. Pedicel length was also highly dependent on temperature and PPFD. As mean temperatures increased or as the temperature differential changed towards a differential of 12 °C, and as PPFD decreased, pedicel length increased significantly. Hip diameter was more sensitive to environmental changes than waist diameter and thus had a greater effect on flower shape. This work indicates to growers that they can enhance Sandersonia flower quality by producing more desirably shaped flowers with short pedicels through maintaining relatively low (less than 23 °C) mean temperatures, minimising temperature differentials, and maintaining irradiance levels as high as possible.     

Enhanced thermotolerance of the vegetative part of MT-sHSP transgenic tomato line

Transgenic tomato (Lycopersicon esculentum Mill.) line was developed that over-expressed tomato MT-sHSP to study the role of MT-sHSP in imparting tolerance to high temperature to vegetative part. T₀ and T₁ generation lines of transgenic tomato and wild type were used for molecular and physiological characterization for thermotolerance. Plants were grown at optimum (26/20 °C) and two supra-optimum, viz. ST₁ (30/22 °C) and ST₂ (32/25 °C) day/night temperature cycles. Plant height, leaf area, as well as assimilation (P_N), leaf cell membrane thermostability (LCMT), and night respiration were recorded. The T₀ and T₁ lines showed increased thermotolerance under both supra-optimum temperatures. Moreover, P_N did not limit the vegetative and reproductive growth under elevated temperatures. After exposure to heat-shock, the wild type and during growth at elevated temperature the transgenic line accumulated proline in the leaf, though more in wild type. Results of the LCMT clearly showed that wild type possessed more heat acclimation potential during growth at elevated temperature than the transgenic line. Further, the expression of MT-sHSP by both the transgenic and wild type under optimum temperature and after heat-shock, respectively, showed that transformation of tomato genome for high temperature tolerance could be a reality; the transformation by itself did not affect the expression of the gene for MT-sHSP. We found that a correlation exists between thermotolerance and both T₀ and T₁ generations of tomato plants and also inheritance of the MT-sHSP gene in T₁ progeny. It is assumed that MT-sHSP is just not expressed by plants under heat-shock, but has a unique function involved in thermotolerance. Specific role of the MT-sHSP in thermotolerance, however, need to be ascertained by in vitro studies.     

Guide for Authors

Bromeliad growers report severe leaf quality problems for Aechmea cultivars grown under commercial greenhouse conditions. In this research, a leaf damage ‘sensitive’ and ‘insensitive’ A. cultivar were compared for their C-metabolism and hydrophysiology under these greenhouse conditions. Stomata opening index indicated CAM (Crassulacean acid metabolism) for both cultivars, with 45–50% open stomata around 4:00 a.m. and 5–10% in the afternoon. Malic and citric acid were the major organic acids present in the leaves, with diurnal malic acid accumulation during the night. The leaf damage ‘sensitive’ cultivar showed higher malic acid accumulation than the ‘insensitive’ cultivar (200 μmol/gfw versus 170 μmol/gfw). Leaf osmotic potential and turgor pressure were linked with diurnal malic acid fluctuations: organic acid accumulation during the night generated high leaf turgor pressures (up to +0.86 MPa).A leaf damage sensitivity test was designed and confirmed previous experiences of several bromeliad growers. Both cultivars showed higher leaf damage percentages (98 and 78%) when leaf malic acid accumulation was high. We attribute this elevated sensitivity to lethal turgor pressures and consider them to be caused by the water capturing mechanism generated by high organic acid accumulation. Under current greenhouse conditions, this water capturing mechanism can be disastrous for plant leaf quality. Especially, under high relative humidity, hindering plant transpiration and stimulating water uptake by dew formation, leaf quality could be endangered.     

Production of high quality Ardisia plants by stem tip cuttings

To produce commercially acceptable Ardisia plants, stem tip cuttings from mature plants were rooted and forced in greenhouses. Ten centimeter long cuttings were either treated with 200 ppm 1-naphthalene acetic acid (NAA) for 2 h, 2000 ppm indole-3-yl-butyric acid (IBA) for 10 s, or 0.5 and 1.0% IBA powder prior to sticking them in the rooting medium. Rooting percentage at 45 days exceeded 76% with 2000 ppm IBA treatment which was a significant increase over non-treated control. Rooted cuttings developed into three types of plants: those forming only vegetative shoots without flowers, those forming reproductive shoots with flowers, and those forming both vegetative and reproductive shoots. The ideal plant produced only vegetative shoots when rooted cuttings were transplanted into pots. About 50% rooted cuttings were forced to finish, producing 31 or 40% of high quality plants when rooted cuttings with vegetative shoots were grown in a greenhouse (GH) at temperatures higher than 21/19 °C (day/night) in 1995 or 21/18 °C GH in 1997, respectively. This method shortened the total production time to less than 2 years as compared to 4 years when starting from seeds.     

Application timing and concentration of abscisic acid affect the quality of ‘Redglobe’ grapes

Summary

The plant hormone abscisic acid (ABA), the concentration of which increases in grape berry skins at the onset of maturation (veraison), appears to be involved in the regulation of anthocyanin accumulation. Preliminary tests suggested that exogenous applications of ABA could improve grape berry colour, but its high cost precluded the development of commercial applications. Recently, a lower-cost ABA production method was developed, which led to the evaluation of different concentrations of ABA, applied at or around veraison, on the quality of ‘Redglobe’ grapes. In two of three years of tests, several ABA treatments enhanced the anthocyanin content of grape skins. ABA, applied at approx. 300 mg l^–1 at veraison, may be required to reliably increase pigmentation, and improve the colour of ‘Redglobe’ grapes. Although the total anthocyanin content was increased by ABA treatment, anthocyanin composition was not affected. Applications of ABA had few effects on fruit size or composition, although they did cause fruit softening, which is undesirable. A secondary objective of this study was to determine how the anthocyanin content of berry skins affected berry colour characteristics. Strong curvilinear relationships between anthocyanin content and lightness and hue showed that these colour characteristics were saturated by anthocyanin contents over 0.02 mg cm^–2 of skin. These data suggest that colour measurements may be needed to evaluate the effect of cultural practices on colour in table grapes.     

紫外光对‘北陆’越橘转色果花青苷积累、关键酶活性及其基因表达的影响

以3年生‘北陆’越橘为试材,对发育过程中果实花青苷含量及合成酶活性变化进行研究,并进一步利用3种紫外光（UV-A、UV-B、UV-C）分别照射转色期越橘果实,测定了果实中花青苷等酚类物质的含量、生物合成酶活性及其基因表达,从底物、酶活性、基因转录水平阐明了不同紫外光对花青苷生物合成途径的影响。3种紫外光都明显诱导了果实花青苷的积累,特别是UV-C照射后果实花青苷含量提高了2.36倍。UV照射显著诱导果实PAL、UFGT酶活性升高,VcPAL、VcUFGT转录增强,但同时也抑制了DFR酶活性及基因表达,其中PAL、UFGT酶活性与花青苷的积累极显著正相关（r = 0.807**,r = 0.894**）,而DFR酶则相反（r =–0.854**）。结果表明,UV照射能诱导丙苯氨酸途径中响应紫外处理的一些关键基因的转录激活（如VcPAL,VcUFGT）或抑制（如VcDFR）,并影响相应酶活性的变化,促使花青苷等酚类物质的积累。     

查耳酮异构酶在葡萄果实发育过程中的表达及亚细胞定位研究

以‘赤霞珠’葡萄果实为材料,利用制备好的葡萄查耳酮异构酶(CHI;EC 5.5.1.6)CHI多克隆抗体,采用RT-PCR、蛋白质印记杂交和胶体金免疫电镜技术对葡萄果实发育过程中CHI基因、蛋白表达及亚细胞定位进行了研究。结果表明:CHI基因和蛋白的表达随着果实发育有着明显的变化,在果实发育早期和转色期表达量较高,同果实中总类黄酮含量的积累变化一致;胶体金免疫定位显示在果实发育早期CHI主要分布于葡萄果皮细胞的细胞质中,在液泡及质体中也有少量分布;在转色期主要定位于细胞质、质体和细胞核中,其数量明显增多。在成熟果实中,主要存在于细胞质,少量存在于质体和液泡中。     

Effect of temperature on seed and fruit development in three mango (Mangifera indica L.) cultivars

The effect of diurnal maximum/minimum (20/10 or 25/15 °C) temperatures on seed and fruit development of ‘Irwin’, ‘Kensington’ and ‘Nam Dok Mai’ mangoes (Mangifera indica L.) was studied in a controlled-environment glasshouse. Exposure to low temperatures (20/10 °C day/night) 3 days after hand pollination significantly increased the percentage of stenospermocarpic fruit (nubbins), in which embryos were aborted at some stage during early fruit development. There were significant differences between cultivars in the percentage of nubbins produced out of the total fruit set following overnight exposure to 10 °C with 21% for ‘Nam Dok Mai’, 11% for ‘Kensington’ and 3% for ‘Irwin’. At 45 days after pollination, nubbin fruits were much smaller in size and weighed ca. 50% less than normal fruits. The lower percentage of nubbin fruits in ‘Irwin’ implies a greater adaptation to cool temperatures by this cultivar during fruit set and early embryo development.     

Environmental factors affecting flowering of rice flower (Ozothamnus diosmifolius,Vent.)

Rice flower (Ozothamnus diosmifolius, Vent.), native to east Australia, is a spring flowering perennial shrub. It is a new cut flower plant, recently introduced into cultivation in Australia and in Israel. Its response to environmental conditions, which affect growth and flowering, are not yet known. The purpose of the present study was to evaluate the effects of growth temperature, photoperiod and total solar energy on flowering. Experiments were conducted with plants of the cv. Cook’s Snow White. Plants were grown in three cycles under controlled conditions in the phytotron, at four day/night temperature regimes: 17/9, 20/12, 23/15 and 26/18°C. Two photoperiods — short day (SD) of 10 h natural day light and long day (LD) of 10 h natural light plus 10 h incandescent light — were employed. High temperatures enhanced vegetative growth but blocked flowering under both LD and SD. Under medium–moderate temperatures plants were absolute LD plants and did not flower under SD conditions. Under lower temperatures plants flowered under both LD and SD, but SD delayed flowering. High total solar radiation under LD did not affect flowering time but greatly promoted the number of flowering stems.     

Interaction of temperature and vegetative flush maturity influences shoot structure and development of lychee (Litchi chinensis Sonn.)

Potted lychee trees (cv. Tai so) of varying vegetative flush maturity were grown under a range of temperature regimes and monitored for subsequent shoot structure and development. A combination of low temperature (15/17 or 18/13 °C day/night) and high vegetative flush maturity was necessary for floral initiation to occur. Exposure to high temperatures (28/23 °C) invariably resulted in the production of vegetative shoots, irrespective of flush maturity. Strong floral initiation was marked by the emergence of terminal panicles and accompanying axillary panicles. A decrease in vegetative flush maturity or increase in temperature (e.g. 23/18 °C) resulted in a decrease in axillary shoot formation and the production of several intermediate shoot structures. These included leafy panicles, stunted panicles, partially emerged buds and non-emergent swollen buds, often produced on the same tree. At 23/18 °C, closer synchronisation of initial flush maturity was required for the production of a consistent shoot-type. Trees with synchronised mature flushes (I-2) at 23/18 °C resulted in the production of swollen terminal buds. Healthy trees were maintained in this state for at least 11 months. These results indicate that both temperature and flush maturity can influence subsequent shoot structure of lychee. In the absence of either a strong floral temperature (18/13 °C) or strong vegetative temperature (28/23 °C), slight differences in initial flush maturity have greater impact on the type of emerging shoot formed.     

Flowering of Primula malacoides in response to photoperiod and temperature

Primula malacoides Franch. ‘Prima Lilac’ was grown at 16 or 20 °C in combination with short days (SD, 8 h) or long days (LD, 16 h). In addition to uninterrupted growing conditions, plants within each temperature were moved at weekly intervals to the other photoperiod and left until termination. Temperature, but not photoperiod, significantly affected the rate of development from start of treatments (51 days from seeding) to 2 mm visible flower bud (VB). At 16 °C, VB averaged 30 days and at 20 °C, 48 days. Time to flower (first horizontal petals) at 16 °C increased from 56 to 64 days as SD increased from 1 week to continuous conditions while LD decreased time to flower from 64 to 56 days. Time to flower at 20 °C varied from 73 to 87 days with additional SD exposure resulting in slower and LD in faster flowering. These observations of the flowering response in ‘Prima’ are contrary to the photoperiodic classification of P. malacoides as a SD plant.     

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.     

Freezing temperature treatment induces bud dormancy in ‘Granny Smith’ apple shoots

One-year-old, ‘Granny Smith’ apple (Malus × domestica Borkh.) shoots were selected randomly from commercial orchards in 1999 and 2003, cold stored at temperatures between 1 and 13 °C for varied periods, following a 12/12 h freezing temperature pre-treatment of −1/13 °C (supposedly non-chilling temperatures) for 1 or 2 weeks. The rate of budburst, determined after forcing at 25 °C, was used to follow the progression of bud dormancy. The freezing pre-treatment clearly enhanced (deepened) bud dormancy in all experiments. Clearly definable influences due to the presence of leaves during the pre-treatment were not observed on shoots cold stored at chilling temperatures. Trees were defoliated before leaf drop in late summer/autumn under field conditions by hand (2001) and using a spray containing 3% urea and 1.8% zinc sulphate in 2003. In 2001 the first two hand defoliation dates significantly enhanced dormancy, while in 2003 chemical defoliation had no effect. These data indicate that the leaves are not clearly involved in the perception of factors that are responsible for dormancy induction. As with dormancy release it is possible that the perception of induction factors, i.e. low temperatures, and as shown in these data, freezing temperatures, occurs within the buds themselves.     

5–氨基乙酰丙酸促进桃果皮提前着色机制研究

 在‘早白花’桃果实着色前,用300、100和50 mg · L^-1的5–氨基乙酰丙酸（5-aminolevulinic acid,ALA）溶液涂布果实。结果表明：ALA处理可促进果皮提前着色,促使果实提前成熟,且以300 mg · L^-1效果最为明显,比对照早约5 d。ALA处理还显著促进了UFGT、DFR、LDOX和CHS的提前表达,并且基因表达水平与花色素苷的积累趋势具有很好的一致性。据此推测ALA可能参与了桃果实成熟以及花色素苷合成的调控,对果皮花色素苷提前合成具有促进作用。