Transcriptomics of the grape berry shrivel ripening disorder

Plant Molecular Biology - The lower expression at veraison of several ripening master regulators “switch genes” can play a central role in the induction of the berry shrivel ripening...     

Analysis of protein changes during grape berry ripening by 2-DE and MALDI-TOF

Grape berry, a nonclimacteric fruit, during ripening turns from green, hard and acidic to coloured, soft and sweet. Many studies have focused on dynamic changes of mRNA levels, metabolites, sugars or individual proteins, but this is the first report of a proteomic approach applied to the screening of the most prominent variations that take place during berry ripening. Vitis vinifera cv. 'Nebbiolo Lampia' berries were collected at 10-day intervals, starting 1 month after flowering to complete ripe stage; total protein extracts from deseeded berries were separated by 2-DE. A total of 730 spots were detected in the 2-DE gels. 118 protein spots, differentially expressed during berry development, were subjected to MALDI-TOF analysis. Ninety-three of them were identified, corresponding to 101 proteins. The majority of proteins were linked to metabolism, energy and protein synthesis and fate. In comparison to published surveys of major berry proteins, fewer proteins related to stress response and more proteins related to cell structure were differentially expressed. Our data confirm a general decrease of glycolysis during ripening, and an increase of PR proteins in the range of 20-35 kDa. They furthermore suggest that oxidative stress decreases during ripening while extensive cytoskeleton rearrangement takes place in this period.     

Abscisic acid,sucrose, and auxin coordinately regulate berry ripening process of the Fujiminori grape

The aim of this study was to examine the effect of abscisic acid (ABA), sucrose, and auxin on grape fruit development and to assess the mechanism of these three factors on the grape fruit ripening process. Different concentrations of ABA, sucrose, and auxin were used to treat the grape fruit, and the ripening-related indices, such as physiological and molecular level parameters, were analyzed. The activity of BG protein activity was analyzed during the fruit development. Sucrose, ABA, and auxin influenced the grape fruit sugar accumulation in different ways, as well as the volatile compounds, anthocyanin content, and fruit firmness. ABA and sucrose induced, but auxin blocked, the ripening-related gene expression levels, such as softening genes PE, PG, PL, and CELL, anthocyanin genes DFR, CHI, F3H, GST, CHS, and UFGT, and aroma genes Ecar, QR, and EGS. ABA, sucrose, and glucose induced the fruit dry weight accumulation, and auxin mainly enhanced fruit dry weight through seed weight accumulation. In the early development of grape, starch was the main energy storage; in the later, it was glucose and fructose. Sucrose metabolism pathway-related gene expression levels were significant for glucose and fructose accumulation. BG protein activity was important in the regulation of grape ABA content levels. ABA plays a core role in the grape fruit development; sucrose functions in fruit development through two pathways: one was ABA dependent, the other ABA independent. Auxin blocked ABA accumulation to regulate the fruit development process.     

Novel aspects of grape berry ripening and post-harvest withering revealed by untargeted LC-ESI-MS metabolomics analysis

We established a step-by-step, experiment-guided metabolomics procedure, based on LC-ESI-MS analysis, to generate a detailed picture of the changing metabolic profiles during late berry development in the important Italian grapevine cultivar Corvina. We sampled berries from four developmental time points and three post-harvest time points during the withering process, and used chromatograms of methanolic extracts to test the performance of the MetAlign and MZmine data mining programs. MZmine achieved a better resolution and therefore generated a more useful data matrix. Then both the quantitative performance of the analytical platform and the matrix effect were assessed, and the final dataset was investigated by multivariate data analysis. Our analysis confirmed the results of previous studies but also revealed some novel findings, including the prevalence of two specific flavonoids in unripe berries and important differences between the developmental profiles of flavones and flavanones, suggesting that specific individual metabolites could have different functions, and that flavones and flavanones probably play quite distinct biological roles. Moreover, the hypothesis-free multivariate analysis of subsets of the wide data matrix evidentiated the relationships between the various classes of metabolites, such as those between anthocyanins and hydroxycinnamic acids and between flavan-3-ols and anthocyanins.     

Reciprocity between abscisic acid and ethylene at the onset of berry ripening and after harvest

Background  

The ripening of grape berry is generally regulated by abscisic acid (ABA), and has no relationship with ethylene function. However, functional interaction and synergism between ABA and ethylene during the beginning of grape berry ripening (véraison) has been found recently.     

Proteomic analysis of grape berry cell cultures reveals that developmentally regulated ripening related processes can be studied using cultured cells

Background

This work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first.

Methodology/Principal Findings

In this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions.

Conclusions

The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening.     

Sugar uptake by the grape berry: A note on the absorption pathway

Summary ³H-glucose was fed to excised Sultana grape berries via their pedicels for up to 5 hours. Autoradigraphy showed that the label was distributed throughout the fruit within 1 hour. Microautoradiography of tissue sections taken at a number of points showed that within the pedicel the walls of cortical cells had become heavily labelled, suggesting that the cortical cell walls offered a diffusion pathway for the solutes entering the vascular system from the external aqueous solution. Transport along the pedicel was confined to the central vascular tissue with little radioactivity occurring in the cortical cells. Within the pericarp, the vascular bundles and walls of nearby parenchyma cells had become heavily labelled, indicating that the labelled solute was present within the vicinity of cell walls. The general pattern of ³H-glucose accumulation by excised berries was similar to the deposition pattern of ²⁴C-labelled photosynthate within attached fruit.     

Comparative effects of acetylene and ethylene gas on initiation of banana ripening

Bananas were exposed to acetylene or ethylene at 0·01, 0·1 and 1 ml/litre, under high humidity, for 24 h at 18 °C. They were then transferred to an atmosphere of air alone for a further 4 days and during this period the respiration rate of three fruit from each treatment was measured. Ripeness was then assessed by colour score and soluble solids content. All levels of ethylene initiated ripening. Treatment with ethylene induced a climacteric rise in respiration, an increase in the soluble solids content of the pulp and degreening of the peel. All levels of acetylene, except 0·01 ml/litre, induced a climacteric rise in respiration. Fruit treated with acetylene at 1 ml/litre had a similar colour score and soluble solids content to those ripened by exposure to ethylene. Fruits treated with acetylene at 0·1 ml/litre had a lower soluble solids content and their peel remained green. Treatment with acetylene at 0·01 ml/litre failed to initiate ripening. Sensory evaluation of fruit ripened by acetylene at 1 ml/litre indicated that the acetylene treated fruit ripened slightly more slowly. When compared at the same stage of ripeness fruits from the two treatments were equally palatable.     

膨大期果面喷施IAA对桃成熟期果实性状和相关基因表达的影响北大核心CSCD

为明确桃果实发育后期外源生长素对果实成熟的影响及更深入的认识生长素调控果实发育与成熟的机制,本研究以桃品种‘小白凤’为试材,采用不同浓度的外源IAA(200,10,0.1μmol/L)喷施第2次快速膨大期的桃果实,在处理后10,20,30 d分别取样,分析桃果实的硬度,糖组分(蔗糖、葡萄糖、果糖、山梨醇),果胶,纤维素含量以及乙烯释放量的变化,并对200μmol/LIAA处理后30 d及对照组的果实进行了转录组测序分析,深入认识生长素调控桃果实成熟的作用和机理。结果表明,(1)IAA处理后30 d时,200μmol/L处理组桃果实果肉硬度与对照组相比显著增加,成熟期平均延迟5 d,而10,0.1μmol/LIAA处理组与同期对照组间无显著差异。(2)果肉蔗糖含量在0.1,200μmol/LIAA处理后30 d显著低于同期对照组,在10μmol/LIAA处理后30 d时与对照组无显著差异;葡萄糖、果糖和山梨醇含量在各浓度IAA处理后30 d时多与对照组间无明显差异。(3)在处理30 d时,果肉中可溶性果胶含量在0.1μmol/L IAA处理组与对照组无显著差异,在10,200μmol/LIAA处理组均显著低于对照组,此时不同浓度IAA处理桃果肉中不可溶果胶和纤维素含量均与对照组之间无显著差异。(4)在IAA处理后10,20,30 d时,果实乙烯释放量均表现为0.1,10μmol/L处理与对照组无显著差异,200μmol/L处理组显著低于对照组。(5)转录组数据显示,200μmol/L IAA处理与对照组中共存在86个差异表达基因,KEGG分析发现其中有6个与果实发育成熟密切相关的代谢途径中多个基因的表达谱发生显著变化,且变化趋势与已测定的生理指标数据吻合。综合分析表明200μmol/L的外源IAA处理第2次快速膨大期的桃果实能够延缓果实成熟的进程。     

The hormone content of ripening grape berries and the effects of growth substance treatments

Berries on field-grown Vitis vinifera cv. Doradillo were treated at different times during stage II with benzothiazole-2-oxyacetic acid or 2-chloroethylphosphonic acid, and measurements were made of their growth and hormone content. The concentration of ethylene was low during stage II and declined as berries ripened. Both 2-chloroethylphosphonic acid and benzothiazole-2-oxyacetic acid caused increases in ethylene concentration, yet they had varying effects on ripening: the former applied at the start of stage II and the latter applied 1 week before the end of stage II delayed ripening, while 2-chloroethylphosphonic acid applied at the end of stage II hastened ripening.     

叶片黄化对葡萄生理和果实品质的影响

为了探究叶片黄化对葡萄生理和果实生长的伤害机制,试验以生长在河南省荥阳市蔡寨村表现黄化症状的4年生‘阳光玫瑰’成龄树为试材,通过测定并分析黄化植株与正常植株的茎叶生长量、叶绿素含量、光合特性、叶绿素荧光参数、叶片解剖结构以及果实品质和产量的变化特征,考察叶片黄化后对葡萄生长量、光合指标、叶绿素荧光特性以及果实品质的影响,以期为葡萄开展黄化矫正栽培提供参考依据。研究表明,(1)黄化植株叶片的叶绿素含量、叶片大小、叶片重量、新梢长度、枝条粗度均显著降低;(2)黄化植株叶片的净光合速率(P_n)、蒸腾速率(T_r)、气孔导度(G_s)显著下降,而胞间CO₂浓度(C_i)显著上升;(3)黄化植株叶片的叶绿素荧光参数初始荧光(F_o)、光下最小荧光产量(F_o′)、最大荧光产量(F_m)、相对电子传递速率(ETR)、实际光合量子产量Y(Ⅱ)均比正常植株显著降低;(4)黄化植株叶片厚度、栅栏组织厚度和栅海比比正常植株显著升高,上表皮厚度和...     

A DIGE-based quantitative proteomic analysis of grape berry flesh development and ripening reveals key events in sugar and organic acid metabolism

Grapevine (Vitis vinifera L.) is an economically important fruit crop. Quality-determining grape components, such as sugars, acids, flavours, anthocyanins, tannins, etc., are accumulated during the different grape berry development stages. Thus, correlating the proteomic profiles with the biochemical and physiological changes occurring in grape is of paramount importance to advance the understanding of the berry development and ripening processes. Here, the developmental analysis of V. vinifera cv. Muscat Hamburg berries is reported at protein level, from fruit set to full ripening. A top-down proteomic approach based on differential in-gel electrophoresis (DIGE) followed by tandem mass spectrometry led to identification and quantification of 156 and 61 differentially expressed proteins in green and ripening phases, respectively. Two key points in development, with respect to changes in protein level, were detected: end of green development and beginning of ripening. The profiles of carbohydrate metabolism enzymes were consistent with a net conversion of sucrose to malate during green development. Pyrophosphate-dependent phosphofructokinase is likely to play a key role to allow an unrestricted carbon flow. The well-known change of imported sucrose fate at the beginning of ripening from accumulation of organic acid (malate) to hexoses (glucose and fructose) was well correlated with a switch in abundance between sucrose synthase and soluble acid invertase. The role of the identified proteins is discussed in relation to their biological function, grape berry development, and to quality traits. Another DIGE experiment comparing fully ripe berries from two vintages showed very few spots changing, thus indicating that protein changes detected throughout development are specific.     

Movement of the grape berry moth, Endopiza viteana: displacement distance and direction

Abstract. Mark–release–recapture is used to quantify displacement by adults of the North American grape berry moth, Endopiza viteana Clemens (Lepidoptera: Tortricidae) under field conditions. Moths marked with fluorescent dust are released eight times in the centre of a vineyard over 2 years, and recaptured using pheromone traps and interception traps. In vineyards, male moths are recaptured an average of 13.8 ± 0.8 m from the release site (maximum 58.2 m), whereas female displacement is similar with average flight distances of 11.4 ± 6.7 m (maximum 41.2 m). Increasing wind speed during moth flight activity periods suppresses displacement by both sexes, and females are less likely than males to fly in winds above 0.6 m s^?1. The majority of males are recaptured upwind from the release site or at a tangent to the overall mean wind direction when responding to pheromone traps, whereas female moths trapped in interception traps exhibit a large variability in direction from the release point. Releases of marked moths in woods adjacent to a vineyard demonstrates interhabitat movement by E. viteana males and by a single female. The average maximum displacement by males during interhabitat movement is 105.4 ± 3.9 m, significantly greater than the average maximum of 39.7 ± 6.7 m inside the vineyard habitat.