Modification of tomato taste in transgenic plants carrying a thaumatin gene from Thaumatococcus daniellii Benth

Fruit taste is an important component of fruit quality, but its genetic basis is complex, making it difficult to alter by plant breeding. Thaumatin is a sweet‐tasting, flavour‐enhancing protein produced by fruits of the African plant Thaumatococcus daniellii Benth. Agrobac‐terium‐mediated transformation was used to produce two transgenic tomato lines expressing biologically active thaumatin in fruits. Transgenic tomato fruits from the T₂ plant generation were sweeter than the controls and possessed a specific aftertaste as determined by sensory evaluation. These results demonstrate that transgenic expression of thaumatin could be useful for modifying tomato fruit taste, especially in breeding lines possessing poor fruit taste, such as those carrying a non‐ripening (nor) mutation.     

Chilling-injury of harvested tomato (Solanum lycopersicum L.) cv. Micro-Tom fruit is reduced by temperature pre-treatments

Heat-shocks were used to reduce the development of chilling injury symptoms during ripening of tomato fruit (Solanum lycopersicum L. cv. Micro-Tom). Mature green tomatoes were immersed in 30-50 °C water for 3-9 min before being chilled at 2.5 °C for 0, 0.5, 1, 2, 3, or 14 days, and then held at 20 °C for an additional 7-14 days. The affect of both heat-shock and chilling treatments were independent of fruit weight. Measured at 20 °C after 14 days of chilling, fruit exposed to 40 °C for 7 min exhibited reduced chilling injury symptoms, as measured by their advanced ripening score and decreased rate of ion leakage into an isotonic 0.2 M mannitol solution. Reduced rates of leakage from the symplastic compartment probably contributed to the 2-fold decrease in the amount of ions in the apoplastic space, when compared to the control. A subsequent paper will report the results of metabolic profiling of Micro-Tom tomato fruit subjected to treatments that significantly decreased their development of chilling injury symptoms.     

柑橘CsaccA基因功能鉴定

为改良柑橘果实品质,以一个推测与柑橘果实品质相关的CsaccA基因为研究对象进行功能鉴定。通过该基因过表达转化Micro-Tom番茄,利用PCR和GUS染色鉴定,植株表型观察和生理生化指标测定等方法系统研究CsaccA基因功能。CsaccA基因显著促进植株的生物量,转基因植株根茎叶生物量均显著高于对照,植株生长速度显著快于对照。转基因植株果实提前转色,果实成熟期明显提前。转基因果实品质分析显示,番茄果实主要的糖组分葡萄糖和果糖含量显著增高,主要的酸组分柠檬酸含量显著降低。CsaccA基因有提升果实糖酸比的功能,可为果实品质改良提供参考。     

Defense responses of tomato fruit to exogenous nitric oxide during postharvest storage

Nitric oxide (NO), an important signalling molecule, has shown diverse physiological functions in plants. We investigated physiological responses of harvested tomato fruit (Solanum lycopersicum cv. Ailsa Craig, AC) to NO treatment. NO released by 1 mM sodium nitroprusside (SNP) aqueous solution could effectively retard pericarp reddening of tomato fruit, suppress ethylene production, and influence quality parameters during storage. The activity of antioxidant enzymes in NO-treated tomato fruit was higher in the late storage period compared to the control. RT-PCR analysis showed that expression of six genes related to fruit ripening was regulated by NO treatment, resulting in an increase in resistance of tomato fruit to gray mold rot caused by Botrytis cinerea. Our results demonstrated that application of NO could be a potential method for treating harvested fruit in order to delay ripening, maintain quality and enhance resistance of fruit to fungal pathogens.     

Mode of inheritance of Alcobaca,a tomato fruit-ripening mutant

Summary Alcobaca, an abnormally ripening tomato mutant, was crossed with the normal cultivar Rutgers. Examination of F₁, F₂ and BC₁ populations indicated that a single recessive gene controls the mutant phenotype. No maternal effect was detected.     

Molecular characterization of two banana ethylene signaling component MaEBFs during fruit ripening

EIN3 Binding F-box protein (EBF) is an essential signaling component necessary for ethylene response. However, little information is available on EBF genes during banana fruit ripening. Two EBF genes designated MaEBF1 and MaEBF2 were isolated and characterized from banana fruit. Subcellular localization analysis showed that MaEBF1 and MaEBF2 were both nuclear proteins. Expression of MaEBF1 and MaEBF2 in fruit with four ripening characteristics revealed that MaEBF2 was enhanced by ethylene during fruit ripening, while MaEBF1 changed only slightly. Moreover, the MaEBF2 promoter was activated after ethylene treatment, further supporting its involvement in fruit ripening. More importantly, MaEBF2 was shown to physically interact with MaEIL5, using yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. Together, these results suggest that MaEBF may be involved in banana fruit ripening, at least partly via interaction with MaEIL5. Our findings expand our understanding of the regulatory network of ethylene signaling cascade in banana fruit ripening.     

Magnetic resonance imaging provides spatial resolution of Chilling Injury in Micro-Tom tomato (Solanum lycopersicum L.) fruit

Magnetic resonance imaging (MRI) was used to monitor internal changes in harvested tomato (Solanum lycopersicum L. cv. Micro-Tom) fruit. Measurements of ethylene evolution, respiration, and ion leakage indicated that the fruit developed chilling injury (CI) after storage at 0 °C. Unlike these measurements, MRI provided spatially resolved data. The apparent diffusion coefficient (ADC), which is an indication of water mobility in tissues, was calculated from MRIs of the different parts of the fruit. Storage for 1 or 2 weeks at 0 °C caused no difference in the ADCs (D-values) in the pericarp, but it did lead to higher values in the inner tissues i.e., the columella and locular region compared to non-chilled fruit (P < 0.05). Changes in inner fruit D-values after 1 and 2 weeks of chilling at 0 °C were similar to changes in respiration, ethylene production and ion leakage which increased (P < 0.05) compared to the non-chilled controls. Most CI studies of tomato fruit used pericarp tissue. Our data indicate that columella tissue changes occur in response to chilling injury in tomato fruit and suggest that more caution is needed when interpreting data from experiments commonly used to study this phenomenon.     

Postharvest ultraviolet-C irradiation suppressed Psy 1 and Lcy-β expression and altered color phenotype in tomato (Solanum lycopersicum) fruit

Mature green cherry tomato fruit were harvested and treated with ultraviolet-C (UV-C) irradiation at a predetermined dose of 4.2 kJ m⁻², and stored at 18 °C for 35 days. The effects of UV-C treatment on color change, pigment contents, and the expression of major genes involved in carotenoid metabolism, including Psy 1, Pds, Lcy-β, and Lcy-ɛ, encoding phytoene synthase, phytoene desaturase, lycopene β-cyclase and lycopene ɛ-cyclase, respectively, were examined. The UV-C treated fruit developed a pink red color in contrast to the normal orange red color of control fruit. Lycopene accumulation during ripening in UV-C treated fruit was significantly inhibited but its final content was not affected. However, both accumulation and final content of β-carotene were significantly suppressed in UV-C treated fruit. The lower content of β-carotene, leading to a higher lycopene to β-carotene ratio, is probably responsible for the altered color phenotype in UV-C treated fruit. Psy 1, a major gene involved in lycopene synthesis was inhibited by UV-C irradiation. Significantly suppressed expression of Lcy-β gene was also observed in UV-C treated fruit. Thus it is possible that the lower transformation from lycopene to carotenes contributed to the relatively stable content of lycopene.     

miR172超表达载体的构建及其在micro-Tom番茄中的遗传转化

为了进一步阐明miRNA在番茄不同生长发育阶段尤其是在果实成熟阶段的调控途径,利用聚合酶链式反应(PCR)从番茄(Solanum lycopersicum)的cDNA克隆出miR172基因核心片段,将该基因片段,以及pCAMBIA1300-221载体通过特异性的限制性内切酶双酶切,然后将miR172片段正向连接到载体上,转化大肠杆菌Trans5α,鉴定重组质粒正确后,利用冻融法将重组载体转入农杆菌EHA105中。再次回转大肠杆菌验证获得的重组载体,通过PCR以及双酶切鉴定是否符合预期结果,将测序结果与NCBI上的基因序列相比较,同源性高达100%。结果成功构建了适用于番茄农杆菌遗传转化的植物表达载体,接下里进行转基因植株的培育和鉴定。为通过miR172基因进一步研究果实成熟衰老机理奠定了物质基础。     

The manipulation and modification of tomato fruit ripening by expression of antisense RNA in transgenic plants

Summary The common cultivated tomato (Lycopersicon esculentum Mill.) provides a major focus for improvement of crop quality through genetic engineering. Identification of ripening-related cDNAs has enabled the modification of specific aspects of ripening by manipulating gene expression in transgenic plants. By utilizing antisense RNA to modify expression of ripening genes, we have inhibited the production of the cell wall-metabolising enzymes polygalacturonase and pectinesterase and created transgenic plants that contain, effectively, single, targeted mutations affecting these genes. Furthermore, this approach has been used with previously unidentified cDNA clones to enable both functional identification and manipulation of genes involved in ethylene production (ACC oxidase) and carotenoid biosynthesis (phytoene synthase). The use of antisense RNA targeted to specific genes to alter ripening phenotypes and improve commercial utility of fruit by affecting shelf-life, processing characteristics and nutritional content is discussed.We have used the extreme ripening-impaired mutant, ripening inhibitor (rin) to identify additional genes implicated in the ripening process. This approach has resulted in the cloning of several novel ripening-related mRNAs which are now being studied by antisense experiments. This may enable identification and manipulation of additional genes involved in processes such as softening, flavour and aroma generation and susceptibility to pathogens.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - PE pectinesterase - PG polygalacturonase - SAM S-adenosyl methionine - SARs scaffold attachment regions     

Mode of action of nitric oxide in inhibiting ethylene biosynthesis and fruit softening during ripening and cool storage of ‘Kensington Pride’ mango

The mode of action of nitric oxide (NO) in inhibiting ethylene biosynthesis and fruit softening during ripening and cool storage of mango fruit was investigated. Hard mature green mango (Mangifera indica L. cv. ‘Kensington Pride’) fruit were fumigated with 20 μL L⁻¹ NO for 2 h at 21 °C and allowed to ripen at 21 ± 1 °C for 10 d, or stored at 13 ± 1 °C for 21 d. During ripening and cool storage, ethylene production and respiration rate from whole fruit were determined daily. The 1-aminocyclopropane-1-carboxylic acid (ACC) content, activities of ACC synthase (ACS), ACC oxidase (ACO), and fruit softening enzymes such as pectin esterase (PE), endo-1,4-β-d-glucanase (EGase), exo- and endo-polygalacturonase (exo-PG, endo-PG) as well as firmness and rheological properties of pulp were determined at two- and seven-day intervals during ripening and cool storage, respectively. NO fumigation inhibited ethylene biosynthesis and respiration rate, and maintained higher pulp firmness, springiness, cohesiveness, chewiness, adhesiveness, and stiffness. NO-fumigated fruit during cool storage and ripening had lower ACC contents through inhibiting the activities of both ACS and ACO in the fruit pulp. NO-fumigated fruit showed decreased activities of exo-PG, endo-PG, EGase, but maintained higher PE activity in pulp tissues during ripening and cool storage. In conclusion, NO fumigation inhibited ethylene biosynthesis through inhibition of ACS and ACO activities leading to reduced ACC content in the fruit pulp which consequently, reduced the activities of fruit softening enzymes during ripening and cool storage.     

Inheritance of shelf life and other quality traits of tomato fruit estimated from F<Subscript>1</Subscript>’s,F<Subscript>2</Subscript>’s and backcross generations derived from standard cultivar, <Emphasis Type="Italic">nor</Emphasis> homozygote and wild cherry tomato

A tomato cultivar with high quality fruit and a long shelf life is a main goal in tomato breeding and it would be achieved using wild germplasm. The objective of this work was to explore the inheritance for fruit quality traits, especially fruit shelf life, in three tomato crosses using a standard Argentinean cultivar (Ca, cv ‘Caimanta’), a ripening mutant (nor, homozygous for the nor gene) of Solanum lycopersicum, and a wild cherry type (Ce, LA1385 of S. lycopersicum var. cerasiforme). The wild parent had a shorter fruit shelf life than the mutant genotype but higher than Ca. When the Ce genotype was analyzed in hybrid combination, the F₁ (Ca×Ce) was similar to the wild genotype for shelf life whereas the F₁ (nor × Ce) had a longer shelf life. Both F₁ crosses and backcrosses to the cherry type genotype had significantly lower fruit weight than the cultivated genotypes but higher than the cherry type parent. In the F₂ analysis, it was found that the inheritance underlying quality traits is complex since non allelic interactions were detected. A significant additive genetic variance was found for fruit shelf life as well as for other fruit quality traits in each cross. The genetic parameters analyzed by mean values and variances in parental, F₁ and F₂ and backcross generations indicated that the cross between the normal ripening cultivar and LA1385 of S. lycopersicum var. cerasiforme offers the best possibility to obtain long shelf life tomato genotypes with good fruit quality.     

Metabolomics of capsicum ripening reveals modification of the ethylene related-pathway and carbon metabolism

Capsicum (Capsicum annuum L. cv. Aries) is a non-climacteric bell-pepper fruit, exhibiting limited ethylene and respiration levels during ripening. In contrast to climacteric fruit, such as tomato which is largely dependent upon ethylene to ripen, the regulation of non-climacteric ripening is still inadequately understood. A metabolomics approach was used to identify differentially abundant compounds between ripening stages with the aim of elucidating metabolic pathways involved in the regulation of non-climacteric ripening. Metabolite profiling using gas chromatography–mass spectrometry (GC–MS) was initially employed to screen potential metabolite differences among three ripening stages (Green, Breaker Red 1 and Light Red). Targeted analyses using liquid chromatography–mass spectrometry (LC–MS) or enzymatic assays were subsequently employed to characterise selected metabolites in more ripening stages. Starch, sugars and their derivatives were significantly modified during ripening which may affect the abundance of some glycolysis intermediates and consequently other metabolic pathways involving amino acids, colour and pungency precursors, and tricarboxylic acid (TCA) cycle intermediates. Furthermore, metabolites closely related to ethylene production such as cysteine and methionine gradually increased between the ripening stages, whereas putrescine significantly decreased during ripening, suggesting that some parts of the ethylene pathway may still be functional in this non-climacteric fruit. Thus, this study which utilised both profiling and targeted metabolomics, has identified a wide range of metabolites which are involved in various biochemical pathways and highlights the overall metabolic shifts during non-climacteric capsicum ripening.     

Pleiotropic effects of male sterility genes in hybrid tomatoes (Lycopersicon esculentum Mill.)

Summary The influence of seven recessive male-sterile alleles on early marketable yield, total marketable yield and average fruit weight in heterozygous F1 hybrid tomatoes was studied. Differences were found between nearisogenic F1 hybrid pairs, in the mutants ms-14, ms-17 and ms-18 for percentage of early marketable yield; in the mutants ms-17, ms-31 and ms-47 for average fruit weight; and in the mutants ms-14, ms-17 and ms-33 for total marketable yield. These differences may be attributed to pleiotropic effects of male-sterility alleles or genes highly linked to these alleles. The direction and the magnitude of the pleiotropic effect on a specific characteristic were found to be dependent on the general genetic background of the parental lines and the specific combining ability of the F1 hybrid. Interactions found in crosses between male-sterile and male-fertile female lines and parental lines suggest that male-sterility alleles affect the general combining ability of female lines. Such differences were demonstrated to be statistically significant for percentage of early marketable yield with mutant ms-18, for average fruit weight with mutants ms-47, and for total marketable yield with mutant ms-17 and ms-33. The absence of uniformity with respect to the pleiotropic effects and occasional deviations in the general combining ability are discussed.