Diffusivity of 1-methylcyclopropene in spinach and bok choi leaf tissue,disks of tomato and avocado fruit tissue,and whole tomato fruit

Gaseous 1-methylcyclopropene (1-MCP) has been widely employed for delaying ripening and senescence of harvested fruit and vegetables; however, details on ingress of gaseous1-MCP in plant tissues, which might contribute to differences in responsiveness of different horticultural commodities to 1-MCP, have not been reported. In this study, we used spinach and bok choi leaves, disks from tomato epidermis, stem-scar and avocado-exocarp tissues, and whole tomato fruit to examine ingress of gaseous 1-MCP. Using a dual-flask system, equilibration of 20 μL L⁻¹ (831 μmol m⁻³) 1-MCP through leaf tissue was reached within 1–2 h, and paralleled 1-MCP transfer through glass-fiber filter paper. For disks derived from fruit tissues, changes in 1-MCP concentrations in the dual-flask system showed anomalous patterns, declining as much as 70% in source flasks with negligible accumulation in sink flasks. The pattern of 1-MCP distribution was markedly different from that of ethylene, which approached equal distribution with tomato stem-scar and avocado exocarp but not tomato epidermis tissues. 1-MCP ingress was further addressed by exposing whole tomato fruit to 20 μL L⁻¹ 1-MCP followed by sampling of internal fruit atmosphere. Tomato fruit accumulated internal gaseous 1-MCP rapidly, reaching approximately 8–9 μL L⁻¹ within 3–6 h at 20 °C. Internal 1-MCP concentration ([1-MCP]) declined around 74 and 94% at 1 and 3 h after exposure, respectively. Ingress was similar at all ripening stages and reduced by 45% in fruit coated with commercial wax. Blocking 1-MCP ingress through stem- and blossom-scar tissues reduced accumulation by around 60%, indicating that ingress also occurs through epidermal tissue. Fruit preloaded with 1-MCP and immersed in water for 2 h retained about 45% of post-exposure gaseous [1-MCP], indicating that 1-MCP is not rapidly sorbed or metabolized by whole tomato fruit. Rapid ingress of gaseous 1-MCP was also observed in tomato fruit exposed to aqueous 1-MCP. Both accumulation and post-exposure decline in internal gaseous [1-MCP] are likely to vary among different fruit and vegetables in accordance with inherent sorption-capacity, surface properties (e.g., waxes, stoma), volume and continuity of gas-filled intercellular spaces, and tissue hydration.     

Identification of quantitative trait loci underlying seed protein and oil contents of soybean across multi‐genetic backgrounds and environments

Seed protein and oil contents are important quantitative traits in soybean. Previously, quantitative trait loci (QTL) associated with seed protein and oil were mostly identified in single genetic background. The objective of this work was to identify QTL and their epistatic effects underlying seed protein and oil contents in three recombinant inbred line populations (two of them used one common female parent) across eight environments by composite interval mapping. Forty QTL underlying protein content and 35 QTL underlying oil content were identified. Among them, nine were universal QTL underlying protein content and four were universal QTL underlying oil content. Epistatic interactions between QTL underlying seed protein/oil and different genetic backgrounds were detected. Different pairs of epistatic interactions were observed in diverse genetic backgrounds across multi‐environments. Common marker intervals were observed to simultaneously underlie seed protein and oil contents with different epistatic interactions. The results in this study suggested that a specific genotype with high oil content and low protein content might significantly affect the selection of soybean lines for high seed protein.     

Improving tomato fruit quality and the european regulatory framework

Tomato fruit quality is determined by a complex set of characters. Fruit used by the fresh market industry must have different attributes than those used by the processing industries. We have determined the major targets of the tomato industry and have isolated from the tomato itself genes which based upon their biochemical function are expected to improve texture and rheology, colour, soluble solids and flavour. Our research is aimed at proving this concept. So far we have demonstrated that polygalacturonase (PG) is a major fruit quality gene providing the processing tomato industry with improved viscosity. Commercial tomato hybrids have been developed in collaboration with Petoseeds (USA) which carry this effect gene. The regulatory system in Europe will be reviewed. Field trials are covered by the EC directive 90/220, which has now been implemented by all countries belonging to the European Union. On the contrary there are no European-wide regulations in place to cover the production of genetically modified plants as food sources. The evolving food regulations will be reviewed and evaluated in the context of competitiveness of the European plant biotechnology industry.     

Resistance of tomato cultivars and progenies to fruit hollowness

Summary In field experiments with fall and winter tomatoes, the incidence of fruit hollowness was found to range from 0 to more than 80%, depending both on the cultivar and the environment. The most resistant cultivars were Bonner Beste and Potentate; the most susceptible were Exhibiton, Moneymaker, E.S. 1, Berco, Cromco and Ailsa Craig. The rate of hollowness of F₁ and F₂ generations of crosses between resistant and susceptible parents suggests that resistance behaves as a partially dominant character governed by a small number of genes. Thus, commercial F₁ hybrids resistant to the disorder are easy to produce. The heritability of the resistance character was found to be rather high.Contribution from The Volcani Institute of Agricultural Research, Bet Dagan, Israel. 1969 Series, No. 1558-E.     

番茄果实总RNA提取与cDNA-AFLP

从番茄果实中提取高质量的RNA是对调控番茄果实品质、生长发育和代谢等相关基因进行分子生物学研究的基础。实验表明,已报道的相关RNA的提取方法,即使利用已经报道的从其它果实中成功提取出高质量RNA的方法,也不能从番茄果实中分离出高质量的RNA。 这主要是由于番茄果实中多糖、多酚类物质及其他次生代谢,在RNA提取过程中与RNA共同沉淀,从而影响RNA的产量和质量。目前,商业化的RNA抽提试剂盒无法直接用于番茄果实RNA的提取。笔者采用4种方法对番茄果实总RNA进行提取,筛选出一种最佳的改进方法。可以从不同时期番茄果实中成功提取高质量的RNA,产量可达401.6--840.2μg/g,通过测定A260/A230和A260/A280 的比值表明,该方法排除了番茄果实中蛋白、多糖、以及多酚等次生物质的干扰,提取到了高质量番茄果实总RNA;1.2%琼脂糖电泳结果表明RNA在整个提取过程中结构完整,未发生明显降解;所提取的RNA用于cDNA-AFLP等到了清晰的表达图谱,这表明其质量完全可满足下一步分子生物学研究。     

Genotype and environmental interaction for fruit quality traits in vintage tomato varieties

Tomato (Solanum lycopersicum L.) is the second most highly consumed vegetable in the world after potato. Traits including physicochemical (lycopene, total titratable acid (TTA), total soluble solids (TSS) and vitamin C), morphological (fruit shape and size) and colors contribute to the overall fruit quality of tomato. The primary objective of the present study was to evaluate vintage tomato varieties representing a wide genetic background for fruit quality including physicochemical, morphological and color traits in multiple environments and to analyze consistency of their performances across locations. In order to achieve this objective, we acquired 44 vintage tomato varieties and evaluated them in five environments (NC, NY, OH in 2009, and NC and OH in 2010). Analysis of the data revealed that there was a significant (p < 0.01) difference among genotypes and environments for all quality traits, Genotype × Environment interaction was significant (p < 0.01) for all quality traits except for TSS. Broad-sense heritability of physicochemical traits ranged from 5.8 % (lycopene) to 35.7 % (TTA) whereas that for morphological traits ranged from 8.1 % (proximal eccentricity) to 97.3 % (fruit shape index external 1), and color from 69.0 % (a*-value) to 99.3 % (b*-value). Pearson’s correlation analysis indicated that estimated lycopene content was negatively correlated with the other physicochemical traits whereas vitamin C, TSS and TTA were positively correlated with each other. Principal component analysis (PCA) based on phenotypic data identified five components explaining at least 82 % of the total variation. Cluster analysis based on phenotypic data revealed the clusters of vintage tomato varieties were not distinct whereas single nucleotide polymorphism data revealed three distinct populations. This information including heritability and correlation coefficients from the present study may be useful to tomato breeding programs to choose germplasm, improve the response to selection and simultaneously improve multiple traits for tomato fruit quality.     

Native fruit tree genetic resources in Japan

The diversity of climate, from subarctic to subtropical, and the complex geological history of Japan have produced a rich biodiversity. The flora includes several hundred species of native woody plants with edible fleshy fruits or nuts. People have eaten them from prehistoric times until about a half century ago. In Hokkaidō and the Ryūkyū Islands nut species had an important role in the diet, but fleshy fruits were also eaten until recently. Only Castanea crenata and a few minor species became domesticated as edible fruit trees in pre-modern times. Recently, Vitis coignetiae, Lonicera caerulea, Akebia quinata, Akebia trifoliata, Stauntonia hexaphylla, and Actinidia arguta have entered small-scale cultivation. The conservation of the germplasm of many of these native species, both in situ and ex situ, is precarious.     

Quantitative trait loci influencing fruit-related characteristics of tomato grown in high-temperature conditions

The objective of this study was to identify molecular markers linked to fruit-related traits in the tomato subjected to high temperatures. In total, 160 F₂ plants derived from a cross between a heat-tolerant breeding line, CL5915-93D4-1-0-3 (Solanum esculentum), and a heat-sensitive wild accession, L4422 (S. pimpinellifolium), were grown in a greenhouse. Six traits including fruit number, fruit weight, brix, seed number, fruit setting, and flower number were scored. The distributions of fruit number, fruit set, flower number, and seed number were skewed towards heat susceptibility which is known to be characteristic of L4422. Polymorphic bands were generated by PCR-derived methods of RAPD, ISSR and AFLP Polymorphism, the segregation ratio, and distribution over the genome of the above 3 markers were compared. Ten linkage groups, ranging 20.6–151.6 cM in size, were constructed with 62 informative markers spanning a total of 776.3 cM. Fruit-related quantitative trait loci (QTLs) were non-randomly distributed in the tomato genome. For the 6 traits investigated, 21 QTLs were dispersed on linkage groups 2–5. The genetic effects of the various QTLs were differently exhibited, in our study we have respectively found from 10.5% to 30.2% of the variation explained by the QTL for flower number (FRN4) and brix (BX2). Thirteen QTL-mapped markers were unique to 1 trait, and 4 markers were linked to more than 1 trait. Among them, QTLs linked to the I868-470 marker had effects on fruit weight and brix, and a significant positive correlation between these 2 traits was noted (r = 0.35, P < 0.05). Thus, the I868-470 marker may have the potential for simultaneous selection of high fruit weight and brix. These markers also allowed us to align genome linkage maps across distantly related species and to reveal the co-localization between these QTLs and major genes.     

Effect of wounding on cell wall hydrolase activity in tomato fruit

Changes in the activity of the cell wall hydrolases – polygalacturonase (EC 3.2.1.15), pectinesterase (EC 3.2.1.11) and β-galactosidase (EC 3.2.1.23) – have been investigated following wounding of tomato fruit pericarp tissue (Lycopersicon esculentum cv. Ailsa Craig). In ripening fruit wounding appears to arrest the further synthesis of polygalacturonase. β-Galactosidase synthesis may also have been arrested in ripening fruit. The level of pectinesterase declined over the first 24 h following harvest, and since this was apparent in both wounded and unwounded tissue may be related to a harvest, rather than a wounding effect. There was a recovery of activity in intact fruit by 48 h after harvest but this seems to be impaired in wounded tissue. In the case of pectinesterase, this observation was extended to examine the changes in isoform profile and it appeared that the decline of this enzyme may be associated with the reduction of one specific isoform — PE2. In contrast to ripening fruit, wounding of fruit at the fully ripe stage appears to have no significant effects on the activities of any of these three enzymes.     

Quantifying lycopene synthesis and chlorophyll breakdown in tomato fruit using remittance VIS spectroscopy

The aim of this study was to increase the understanding of chlorophyll breakdown and lycopene synthesis at a quantitative level in Solanum lycopersicum fruit. To accomplish this, a kinetic model is proposed describing the transition from chloro- to chromoplast. Remittance VIS spectroscopy was used to assess chlorophyll and lycopene levels non-destructively in cocktail and round type tomatoes. Tomatoes were stored at constant temperatures between 4 and 24 °C, or at a stepwise changing temperature between 4 and 16 °C. Chlorophyll and lycopene levels were measured repeatedly over time and used to calibrate a kinetic model that describes how an autocatalytic enzyme system links chlorophyll breakdown to lycopene synthesis, including breakdown of lycopene precursor and lycopene itself. Increasing storage temperatures increased the reaction constant for lycopene synthesis more than that of chlorophyll breakdown for both tomato types. The reaction constants describing chlorophyll breakdown and lycopene synthesis were considerably larger, and the estimated enzyme levels lower for the round type. This allows round tomatoes to quickly resume lycopene synthesis after a cold storage period when enzyme levels are low. Lycopene breakdown was established for the round type while the cocktail type showed lycopene precursor breakdown. Chlorophyll breakdown and lycopene synthesis, as affected by storage temperature and tomato type, is covered well by the model for both tomato types. We hypothesise that the postulated enzyme system, responsible for the direct link between chlorophyll breakdown and lycopene synthesis, is due to STAY-GREEN proteins. Remittance VIS spectroscopy is, in combination with a parameter estimation tool, suited to screen tomato genotypes for intended colour transformation performance, or as tool in chloroplast to chromoplast transition studies.     

Papaya endoxylanase biochemical characterization and isoforms expressed during fruit ripening

Papaya fruit ripening processes involve the coordinated action of several hydrolases that causes cell wall degradation. Endoxylanase participates in xylan or arabinoxylan modifications and its importance has been related to papaya softening. However, endoxylanase has been not fully characterized biochemically and kinetically. Semipurified endoxylanase from ripe ‘Maradol’ papaya fruit had an optimal temperature from 45 °C to 50 °C, a pH optimum of 5.5 against Remazol brilliant blue-xylan (RBB-Xylan) and enzymatic activity remained stable during 36 h at 45 °C. The activation energy of the enzyme was 25.5 kJ mol⁻¹, and the V_max at 32, 37 and 42 °C was 788.9, 888.9 and 1085.6 μg kg⁻¹ s⁻¹, respectively. The K_m did not change as a function of temperature and was measured as 1.8 g L⁻¹ and was within the range reported for other xylanases. Total proteins were extracted from color-break, half-ripe and ripe fruit. A pre-endoxylanase at 63.9 kDa was identified in the color-break fruit and an active endoxylanase at 32.5 kDa that was only found in ripe fruit, when the highest enzymatic activity was obtained. Immunodetection on two-dimensional gel electrophoresis (2DE) protein blots showed three isoforms of the pre-endoxylanase at color-break and ripe stages and, four isoforms in ripe fruit that were absent in color-break fruit. The biochemical and kinetic characteristics of the endoxylanase are crucial to our understanding papaya fruit softening.     

Quantitative trait loci (QTL) affecting sugars, organic acids and other biochemical properties possibly contributing to flavor, identified in four advanced backcross populations of tomato

Although the Advanced Backcross strategy has proven very useful for QTL detection in tomato, it has been used mainly in identifying QTL for agronomic traits such as yield, color, etc. Tomato flavor is an important quality characteristic, yet it has been difficult to assess flavor or traits that affect it. In this study the AB-QTL strategy was applied to four advanced backcross populations to identify QTL for biochemical properties that may contribute to the flavor of processed tomatoes, such as sugars and organic acids. A total of 222 QTL were identified for 15 traits, including flavor as assessed by a taste panel. Correlations of certain biochemicals with flavor and possible methods of assessing and improving flavor are discussed. In particular, QTL with very significant effects associated with the ratio of sugars/glutamic acid, a trait highly correlated with improved flavor, have been identified as good targets for future work in improving the flavor of tomatoes. This revised version was published online in August 2006 with corrections to the Cover Date.     

Determining tomato fruit maturity with nondestructive in vivo nuclear magnetic resonance imaging

Nuclear magnetic resonance (NMR) images were taken of freshly harvested fruit (Lycopersicon esculentum cv. Castlemart). Measurements were also made of the stage of repeness, rate of carbon dioxide and ethylene production, lycopene and chlorophyll content, density of the pericarp wall, and consistency of the locular tissue. An NMR image at 256 × 256 pixel per image and 16 levels of intensity per pixel clearly showed many structural details of the fruit. Increased intensity was associated with liquefaction of the placental tissue during ripening, while grainines of the pericarp wall was associated with a decrease in wall density. In vivo NMR imaging of mature-green fruit produced images in which differences in maturity could be seen. The long required to produce an image and the cost of operation currently precludes the use of NMR imaging to sort mature-green tomato fruit.     

Inheritance and genetic linkage analysis of a firm-ripening tomato mutant

The tomato cv. ‘Santa Clara’ is widely cultivated among tomato producers in most of the South‐east of Brazil. Recently, some plants of this cultivar were identified with morphological alterations in both vegetative and reproductive organs. These plants showed firm (firme) ripe fruits, slow and delayed ripening. They also had yellow leaves associated with precocious senescence and flowers with pale stigmas. The objective of this work was to determine the genetic model of inheritance for this mutation and to evaluate its effects on shelf life and loss of firmness in mature fruits, as well as analyse the occurrence of genetic relationships between this putative mutant and other pleiotropic mutants. Mutated plants were crossed with the non‐mutant cv. ‘Santa Clara’ and some previously described pleiotropic mutants. Seeds of F₁ and F₂ generations and backcrosses were obtained for the segregation analysis. Morphological characteristics modified by this mutation are governed by a recessive gene with pleiotropic effects. In addition, the test of allelism showed a lack of genetic complementation between the ‘firme’ mutant and lutescent‐2 mapped on chromosome 10. Fruits of the ‘firme’ mutant had a slower rate of softening compared with the cv. ‘Santa Clara’ and its hybrids. The fruit shelf life of the mutant ‘firme’ was significantly superior to the other genotypes. No maternal effect was detected in either qualitative or quantitative characteristics. Based on the data, the mutation ‘firme’ in the cv. ‘Santa Clara’ is located in the region containing the l‐2 locus, which promoted alterations in ripening and post‐harvest physiology of fruits. The mutation ‘firme’ may represent a new allele of the gene lutescent‐2 or a gene linked to physiological events of fruit ripening.     

A method for evaluating fruit abscission potential of grapes and cherry tomato clusters

Abscission is an important postharvest factor for table grapes and cluster cherry tomatoes. There are methods to measure the individual fruit removal force but until now there has been no objective method to study the abscission at a whole cluster level. The working principle of the instrument is that a cluster is attached to a rod which is allowed to fall freely for a pre-determined distance; as the rod stops, the momentum of the berries is converted to detachment force applied to the abscission zone of each berry. The abscission potential can be evaluated as a percentage of fruit detached from cluster under given conditions. The instrument was constructed for robust and adjustable experimental use. It is shown that the abscission percentage for both table grapes and cluster cherry tomatoes was proportional to the free-fall distance. By using the instrument, we showed that the abscission potential of cluster cherry tomatoes was low during the first 2 weeks of storage and that it increased thereafter. Storage of cluster cherry tomatoes at 7 °C resulted in significantly less abscission than at 12 °C, as assessed either by manual shaking or by using the instrument. Treatment of grapes with the cytokinin-like compound forchlorfenthuron at 4 mg L⁻¹ during the early stages of fruit development resulted in significant reduction in abscission potential. These results demonstrate that the method can be used as a reliable experimental tool for objective evaluation of abscission potential.