All-Flesh Tomato Regulated by Reduced Expression Dosage of AFF Through a Promoter SV Mutation

The formation of locule gel is an important process in tomato and a typical characteristic of berry fruit. In this study, we collected a tomato natural mutant that produces all-flesh fruits (AFF) in which the locule tissue remains in a solid state during fruit development. We built genetic populations to fine-map the causal gene of the AFF trait and identified the gene AFF (SlMBP3) as the locus conferring the locule gel formation. We determined the causal mutation as a 416-bp deletion that occurred in the promoter region of AFF and reduced its expression dosage. The 416-bp sequence is highly conserved among Solanaceae species, as well as within the tomato germplasm. Furthermore, with the BC6 NIL materials, we revealed that the reduced expression dosage of AFF did not impact the normal development of seeds but produced unique non-liquefied locule tissue, which was distinct from that of normal tomatoes in terms of metabolic components. We further revealed the importance of AFF gene in locule tissue liquefaction through combined analysis using mRNA-seq and metabolomics. Our findings provide clues to investigate fruit type differentiation in Solanaceae crops and also contribute to the application of the AFF gene in tomato breeding programs.

All-Flesh Tomato Regulated by Reduced Dosage of AFF Provides New Insights into Berry Fruit Evolution

ABSTRACTThe formation of locule gel is not only an important developmental process in tomato but also a typical characteristic of berry fruit. In this study, we collected a tomato material that produces all-flesh fruits (AFF), whose locule tissue remains in a solid state during fruit development. We built genetic populations to fine map the causal gene of AFF trait, investigate the function of AFF gene, and identified it as the causal locus conferring the locule gel formation. We determined the causal mutation as a 416-bp deletion that occurred in the promoter region of AFF, which reduces the expression dosage of AFF. The 416-bp deleted sequence has a high level of conservation among closely related Solanaceae species, as well as in the tomato population. The activity of the 416-bp deletion in down-regulating gene expression was further verified by the relative activity in a luciferase experiment. Furthermore, with the BC6 NIL materials, we reveal that the reduced expression dosage of AFF does not impact the normal development of seeds, while produces non-liquefied locule tissue, which is distinct from that of the normal tomatoes in terms of metabolic components. Based on these findings, we propose that the AFF gene is the core node in locule tissue liquefaction, whose function cannot be compensated by its paralogs TAG1, TAGL1, or TAGL11. Our findings provide clues to investigate fruit type differentiation among Solanaceae crops, and also contributes to the breeding application of all flesh fruit tomatoes for the tomato processing industry.One Sentence SummaryThe sequence deletion that occurred in the cis-regulatory region of AFF—the core node of locule tissue liquefaction determined here—reduced its expression dosage, and produced all flesh fruit tomato.

(470) Lycopene Content of Mini Watermelon Varieties Grown at Four Locations

Mini watermelons are the size of cantaloupes and weigh 1.5 to 3 kg (4 to 8 lbs). Melons of 18 selections were grown in replicated trials in North Carolina, South Carolina, and north and south Florida. Melons were harvested when ripe, and samples of heart and locule tissue were frozen and sent to Lane, Okla. A total of 960 samples, representing 6-12 melons per selection per location, were analyzed for total lycopene content using colorimeter and spectrophotometer methods. Subsamples of `Mohican', `Hazera 6007', `Vanessa', `Petite Treat', and `Precious Petite' were analyzed by HPLC for carotenoid profiles. Total lycopene content ranged from 52 to 108 μg·g-1, depending on variety. Selections were grouped into two levels of lycopene content. The varieties Precious Petite, Petite Perfection, Betsy, Bonny, Petite Treat, Valdoria, Vanessa, Hazera 5133 and 5138, RWT 8149, 8155, 8162 had 60 to 79 μg·g-1 lycopene and the varieties Hazera 6007, 5123, 5109, 5177, Mohican, and Extazy had 80 to 100 μg·g-1. Melons harvested from the Florida locations had more total lycopene than those from North and South Carolina. `Precious Petite' had more β-carotene as a percentage of total carotenoids than other varieties tested. These results indicate that lycopene content is affected primarily by germplasm and also by environment.

Internal Bruising Alters Aroma Volatile Profiles in Tomato Fruit Tissues

Tomato (Lycopersicon esculentum Mill.) fruit, cv. Solar Set, were harvested at the mature-green stage and treated with 50 μL·L-1 ethylene at 20 °C. Individual fruits at the breaker stage (<10% red color) were dropped onto a solid surface to induce internal bruising. Dropped and undropped fruit were stored at 20 °C until red-ripe, at which time pericarp, placental, and locule tissues were excised. Tissues from dropped tomatoes were examined for evidence of internal bruising and all tissues were analyzed for selected volatile profiles via headspace analysis. Individual volatile profiles of the three tissues in bruised fruit were significantly different from those of corresponding tissues in undropped, control fruit, notably: trans-2-hexenal from pericarp tissue; 1-penten-3-one, cis-3-hexenal, 6-methyl-5-hepten-2-one, cis-3-hexenol and 2-isobutylthiazole from locule tissue; and 1-penten-3-one and β-ionone from placental tissue. Alteration of volatile profiles was most pronounced in the locule tissue, which was more sensitive to internal bruising than the other tissues. Changes observed in the volatile profiles appear to be related to disruption of cellular structures.

Chemical Composition and Physical Properties of Pericarp, Locule, and Placental Tissues of Tomatoes with Internal Bruising

`Solar Set' tomatoes (Lycopersicon esculentum Mill.) were harvested at the mature-green stage of development and treated with 50 μL·L-1 ethylene at 20 °C. Breaker-stage fruit were dropped from 40 cm onto a solid surface to induce internal bruising and held along with undropped fruit at 20 °C. At the ripe stage, pericarp, locule, and placental tissues were analyzed for soluble sugars, vitamin C, pigments, titratable acidity, soluble solids content, pericarp electrolyte leakage, extractable polygalacturonase activity, and locule tissue consistency. Bruising significantly affected chemical composition and physical properties of pericarp and locule tissues, but not placental tissue. For bruised locule tissue, carotenoids, vitamin C, and titratable acidity were 37%, 15%, and 15%, lower, respectively, than unbruised fruit. For bruised pericarp tissue, vitamin C content was 16% lower than for unbruised tissue, whereas bruising increased electrolyte leakage and extractable polygalacturonase activity by 25% and 33%, respectively. Evidence of abnormal ripening following impact bruising was confined to locule and pericarp tissues and may be related to the disruption of cell structure and altered enzyme activity.

Internal Bruising Affects Chemical and Physical Composition of Tomato Fruit

Tomato (Lycopersicon esculentum L.) fruits, cv. Solarset, were harvested at the mature-green stage and treated with 50 μL/L ethylene at 20C. Breaker fruits (<10% red coloration) were dropped from 40 cm onto a smooth, solid surface and held along with undropped fruits at 20°C and 85% relative humidity. At table-ripe stage, pericarp, placental, and locular tissue were individually excised and analyzed for total carotenoids, total soluble sugars, soluble solids content, titratable acidity, density (locule tissue), polygalacturonase activity, and electrolyte efflux (pericarp tissue). Internal bruising caused by impact forces significantly affected pericarp and locule tissues, but not placental tissue. For bruised locule tissue, total carotenoids content decreased by 37.1%, vitamin C content by 15.6%, and titratable acidity by 15.3% as compared to control. However, density was increased by 3.0%. For bruised pericarp tissue, vitamin C content decreased by 16.5%, while polygalacturonase activity and electrolyte efflux increased by 33.3% and 24.8%, respectively. The development of abnormal ripening following an impact was confined to locule and pericarp tissues and appears to be related to the disruption of cellular structure and stimulation of enzymic activity.

Molecular Mass Changes in Cell Wall Pectins of Tomato Fruit Locule Tissue in Response to Deesterification

Changes in the gel filtration behavior (apparent mol mass) of cell wall pectic polymers have been observed in a number of ripening fruits, including some that express little or no detectable polygalacturonase (PG). Pectins from tomato (Lycopersicon esculentum, Mill. v. Solar Set) fruit locule tissue show limited depolymerization during ripening, although alkali-soluble polymers are of reduced mol mass relative to water- and chelator-soluble polymers (Plant Physiol. 111:447). This study addressed whether the lower mol mass of alkali-soluble polymers was a consequence of extraction or specific metabolism of these wall polymers. Pectins from sequential water and chelator extractions of ethanol-insoluble solids from mature green tomato locule tissue were subjected to alkaline conditions. The size distribution of both water- and CDTA-soluble pectins treated with weak alkali were downshifted and similar to those extracted directly by weak alkali, indicating structural similarities of the three pectin fractions. Spectrophotometric analysis showed no involvement of β-elimination hydrolysis in the apparent mol mass reduction. The alkali-treated polymers were of greatly enhanced susceptibility to PG-mediated degradation. The alkali-associated changes also occurred in response to pectinmethylesterase hydrolysis. The results indicate that deesterification can strongly influence gel filtration behavior of pectins and may explain the apparent mol mass decreases of pectins in fruits not containing PG.

Pectin Solubilization and Enzymic Activity of Tomato Fruit Locule Tissue Cell Wall

Hydrolytic activities in liquefying locule tissue of mature-green tomato (Lycopersicon esculentum Mill. `Solar Set') fruit were studied in pursuing the understanding of mechanisms involved in the rheological changes occurring in this tissue. Ethanol-insoluble solids (EIS) were prepared with and without enzyme-inactivating treatment. The release of uronic acids from enzymically active EIS incubated under autolysic conditions was 5-fold higher than recoveries from inactive EIS. Uronic acid release was partially inhibited by 1 mm Hg2+. Cell-free proteins extracts from active EIS exhibited hydrolytic activity against inactive EIS. Pectins released from active EIS showed no evidence of main-chain hydrolysis. Neutral sugars recovered as 80% ethanol-soluble products of autolytic reactions included glc, gal, rha, ara, xyl, and man. Gal was recovered at significant higher levels in autolysates of active EIS. Glycosidases present at high activities in locule tissue included α- and β-galactosidases, β-mannosidase, β-arabinosidase, and β-glucosidase. The results confirm our earlier findings that the metabolism of water, chelator, and alkali-soluble pectins in tomato locule tissue involves deglycosylation with no apparent depolymeriation. These changes alone appear to be inadequate in explaining the unique rheological characteristics in locule gel tissue.