Abstract
A number of factors in the isolation of good quality mitochondria from tomato fruit wall tissue have been critically examined. Particles from tomatoes at a number of stages during maturation and senescence were isolated and their ability to oxidise succinate, malate and α-ketoglutarate tested. In general, oxidation rates of the mitochondria and their respiratory control and ADP to oxygen uptake ratios reached a maximum with incipient ripeness and then declined with the onset of senescence. Thiamine pyrophosphate increased the oxidation rate of malate and α-ketoglutarate at all stages and appeared to prevent instances of inhibition probably due to the accumulation of oxaloacetate. Particles from the green side of ‘blotchy’ fruit had characteristics similar to those from normal fruit at the mature green stage, while the red side yielded particles that were similar to those from evenly coloured overripe fruit. Fruit having the dominant allele ‘Never ripe’, which causes extreme slowness in ripening, appeared to have mitochondria comparable in behaviour with those isolated from fruit undergoing normal maturation and senescense.
Resume
Un certain nombre de facteurs intervenant dans l'isolement de mitochondries de bonne qualité à partir des tissus de la paroi du fruit de la tomate ont été examinés. Des particules extraites de tomates à différents stades de maturation et de sénescence ont été isolées et leur capacité d'oxyder le succinate, le malate et l'α-cétoglutarate a été testée.
En général, les vitesses d'oxydation des mitochondries, leur contrôle respiratoire et leur rapport ADP/O fixé atteignent une valeur maximum avec la maturité et puis décroissent avec l'instauration de la sénescence. L'addition de pyrophosphate de thiamine augmente la vitesse d'oxydation du malate et de l'α-cétoglutarate à tous les stades et semble empêcher des inhibitions probablement dues à l'accumulation d'oxaloacétate. Des particules isolées de parties vertes de fruits ‘blotchy’ présentent des caractéristiques similaires à celles extraites à partir de fruits normaux au stade de maturité verte, tandis que le côté rouge fournit des particules semblables à celles provenant des fruits trop mûrs uniformément colorés.
Les fruits possédant l'allèle ‘Never Ripe’ dominant, qui provoque une extrême lenteur dans la maturation, semblent présenter des mitochondries dont le comportement est comparable à celui des mitochondries isolées à partir de fruits subissant une maturation et une sénescence normales.
Zusammenfassung
Eine Reihe von Faktoren, eine gute Qualität von Mitochondrien aus Fruchtschalengeweben von Tomaten zu isolieren, wurde kritisch untersucht. Teilchen von Tomaten einer Reihe von Reife- und Altersstadien wurden isoliert und ihre Fähigkeit geprüft, Succinat, Malat und α-Ketoglutarat zu oxidieren.
Im allgemeinen erreichen die Oxidationsraten der Mitochondrien und ihre Steuerung der Atmung sowie die ADP/O-Aufnahme-Verhältnisse ein Maximum mit beginnender Reife und nehmen ab mit einsetzendem Alter. Thiaminpyrophosphat erhöht die Oxidationsrate von Malat und α-Ketoglutarat in allen Stadien und scheint Fälle von Hemmung zu verhindern, die wahrscheinlich auf eine Anhäufung von Oxalacetat zurückzuführen sind. Teilchen der grünen Seite fleckiger Früchte wiesen Charakteristika auf, die denen normaler Früchte im grünen Reifestadium entsprachen, während die rote Seite Teilchen ergab, die ähnlich denen normaler überreifer Früchte waren. Früchte mit dem dominanten Allel ‘Never ripe’, das ein extrem langsames Reifen hervorruft, scheinen Mitochondrien zu besitzen, die in ihren Eigenschaften denen vergleichbar sind, die aus Früchten normaler Reife und von normalen Alter isoliert wurden.
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Hobson, G.E. Respiratory oxidation by mitochondria from tomatoes at different stages of ripeness. Plant Food Hum Nutr 19, 155–165 (1969). https://doi.org/10.1007/BF01101150
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DOI: https://doi.org/10.1007/BF01101150