Analysis of expressed sequence tags from Actinidia: applications of a cross species EST database for gene discovery in the areas of flavor, health, color and ripening.

BACKGROUND: Kiwifruit (Actinidia spp.) are a relatively new, but economically important crop grown in many different parts of the world. Commercial success is driven by the development of new cultivars with novel consumer traits including flavor, appearance, healthful components and convenience. To increase our understanding of the genetic diversity and gene-based control of these key traits in Actinidia, we have produced a collection of 132,577 expressed sequence tags (ESTs). RESULTS: The ESTs were derived mainly from four Actinidia species (A. chinensis, A. deliciosa, A. arguta and A. eriantha) and fell into 41,858 non redundant clusters (18,070 tentative consensus sequences and 23,788 EST singletons). Analysis of flavor and fragrance-related gene families (acyltransferases and carboxylesterases) and pathways (terpenoid biosynthesis) is presented in comparison with a chemical analysis of the compounds present in Actinidia including esters, acids, alcohols and terpenes. ESTs are identified for most genes in color pathways controlling chlorophyll degradation and carotenoid biosynthesis. In the health area, data are presented on the ESTs involved in ascorbic acid and quinic acid biosynthesis showing not only that genes for many of the steps in these pathways are represented in the database, but that genes encoding some critical steps are absent. In the convenience area, genes related to different stages of fruit softening are identified. CONCLUSION: This large EST resource will allow researchers to undertake the tremendous challenge of understanding the molecular basis of genetic diversity in the Actinidia genus as well as provide an EST resource for comparative fruit genomics. The various bioinformatics analyses we have undertaken demonstrates the extent of coverage of ESTs for genes encoding different biochemical pathways in Actinidia.

Planteose is a short-term storage carbohydrate in Actinidia leaves.

The polyol myo-inositol constitutes 10-20% of soluble carbohydrates in mature leaves of Actinidia deliciosa (A.Chev.) C.F. Liang et A.R. Ferguson var. deliciosa 'Hayward' and A. arguta (Sieb. et Zucc.) Planch. ex Miq. var. arguta. In contrast with other non-structural carbohydrates, myo-inositol concentrations in A. deliciosa leaves increase only slightly during development from sink to source, and are not affected in source leaves by increased sink demand upon fruit set. In mature fruit-bearing leaves myo-inositol concentrations fluctuate diurnally, increasing during the night and declining towards morning, but in plants with less sink demand a diurnal pattern is not observed. In potted A. arguta seedlings subjected to extended dark periods, leaf concentrations of sugars and starch decline rapidly while myo-inositol concentrations are maintained. Labelling studies with 14CO2 revealed that myo-inositol in leaves is not a primary photosynthetic product and is turned over more slowly than other soluble carbohydrates. A suggested role of myo-inositol as a precursor in mucilage synthesis was not substantiated, as radioactivity was incorporated into mucilage more rapidly than into free myo-inositol. Planteose, a trisaccharide comprising sucrose and galactose, incorporated substantial amounts of radioactivity and accumulated to high levels, indicating a role in short-term storage of sucrose. Planteose was synthesised during the day and degraded during the night in a manner that was opposite to that of sucrose while starch and myo-inositol levels remained relatively constant. Planteose has been reported in Cyclamen persicum, ash and sesame seed. This is the first report of planteose in Actinidia, and the first time it has been identified as a major short-term storage carbohydrate in Actinidia leaves.

Frost hardening of Pinus radiata seedlings: effects of temperature on relative growth rate, carbon balance and carbohydrate concentration.

Pinus radiata (D. Don) seedlings were grown for 100 days at day/night temperatures of 10/1, 15/1, 20/1 and 25/1 degrees C, to determine whether temperatures above a threshold of 5 degrees C influence frost hardiness development. Relationships between hardening and relative growth rate, carbohydrate concentration and net carbon balance were also investigated. Seedlings hardened at a nearly constant rate in each treatment, although the rate of hardening was strongly temperature dependent. It increased as the temperature declined, but in a curvilinear fashion. Temperatures below 9.5 degrees C were effective in hardening the seedlings. During the daily temperature cycle, dehardening occurred at temperatures above the threshold, whereas hardening occurred at temperatures below the threshold. The net difference between the two processes determined the development of frost hardiness. The development of frost hardiness was negatively correlated with relative growth rate and positively correlated with the accumulation of starch and sugars. We conclude that frost hardening is a complex process that is causally linked to carbohydrate concentrations.