Chijiwa, H.

Fujishima, H.

Shiraishi, M.

English

JKI Open Journal Systems (Julius Kühn-Institut)

Vitis 47 (3), 191–192 (2008)Research NoteTetraploid sucrose-accumulating grapevinesM. SHIRAISHI, H. FUJISHIMA and H. CHIJIWAFukuoka Agricultural Research Center, Fukuoka, Japan K e y   w o r d s :  HPLC, sucrose accumulator, sugar com-position, tetraploid. Introduction: Fructose and glucose are primary sugars of grape (Vitis spp.), and sucrose presents in trace amounts (WINKLER et al. 1974, SHIRAISHI 1993, DAVIS and ROBINSON 1996). However, some American hybrid cultivars such as 'Bath', 'Buffalo', 'Fredonia' and 'Steuben' contain large amounts of sucrose (LOTT and BARRETT 1967, TAKAYANAGI and YOKOTSUGA 1997; SHIRAISHI 2000). The early ripening and strong, sweet taste of these diploid sucrose-accumu-lating cultivars are of great interest in table grape breed-ing to broaden variability among seedlings. However, the diploid berries are in general smaller than the tetraploid ones, and consumers’ acceptance of large-sized grapes is high. Polyploid breeding has been very successful in Japan since 1970, and the tetraploid cultivars with large berries offer the possibility of creating unique table grapes with a striking appearance (REISCH and PRATT 1996). Many tetra-ploid genotypes have also been introduced by crossbreed-ing and in vitro colchicine treatment of diploid cultivars in our breeding program at Fukuoka Agricultural Research Center, Japan (NOTSUGA et al. 2000). As breeding materi-als, these tetraploid genotypes are valuable additions to the gene pool for developing future table grapes. By screen-ing sugar variants, the tetraploid genotypes were gener-ally found to be hexose accumulators (Fig. 1 A), but some tetraploid sucrose accumulators were observed as follows (Fig. 1 B): 'Buffalo 4X' (TSA-1), 'Steuben 4X' (TSA-2), 'Red Port 4X' (TSA-3), and Seedling 11-7 (TSA-4). In this report we document the origins of TSA-1,2,3 and -4, and describe their agro/biochemical traits between 2006 and 2007.Materials and Methods: Four tetraploid sucrose-ac-cumulating genotypes, TSA-1 to -4 (Vitis labruscana × V. vinifera interspecific hybrids) were grown in a green-house between 2006 and 2007 at the Fukuoka Agricultural Research Center, Fukuoka, Japan. Own-rooted, one vine per genotype was used and trained onto a horizontal trel-lis system. The following agro/biochemical traits were measured during growth: time of full maturity, single berry weight, skin color, soluble solids content (SSC), acidity, total sugar content, and sugar composition. Two to three bunches in each genotype were selected. Three berries per bunch were sampled, and were selected from the top, mid-dle, and bottom of the bunch. After microwave heat treat-ment for 20 s, the juices were extracted from the berries by hand pressing and centrifuged at 5000 × g for 10 min. SSC was measured by a hand refractometer and expressed as °Brix. Acidity was determined by potentiometric titra-tion. Sugar composition was determined using a Shimadzu LC-10A HPLC system equipped with a RID-10A refrac-tive index detector. Juice (1 ml) was diluted to 20 ml with deionized water and filtered through a 0.45 μm filter. The column (Cosmosil Sugar-D, 4.6 × 250 mm, Kyoto, Japan) was operated at 42 ºC. The mobile phase was a mixture of acetonitrile and water (75:25, v/v), and the flow rate was 1.0 ml·min-1. The injection volume was 20 μl.Results and Discussion:  O r i g i n :  TSA-1 to -3 were induced between 1984 and 1990 via in vitro chromosome doubling of the diploid sucrose-accumulating cultivars 'Buffalo', 'Steuben', and 'Red Port', respectively (Fig. 2). Correspondence to: Dr. M. SHIRAISHI, Fukuoka Agricultural Research Center, 587 Yoshiki, Fukuoka 818-8549, Japan. Fax: +81-92-922-4916. E-mail: mikioshi@farc.pref.fukuoka.jpFig. 1: Typical HPLC profiles of sugar composition in tetraploid grape genotypes. A: Hexose accumulator (cv. Kyoho) B: Sucrose accumulator (TSA-4: breeding line 11-7).Fig. 2: Pedigrees of sucrose-accumulating tetraploids, TSA-1, TSA-2, TSA-3 and TSA-4.      : Sucrose accumulator. 192 M. SHIRAISHI et al.TSA-4 was introduced in 1990 from an interspecific cross between 'Olympia' and 'Yoho' (Fig. 2). It is interesting that the occurrence of TSA-4 resulted from successive parental crosses between hexose accumulators. Inheritance of su-crose accumulation among diploid- and tetraploid-grape-vines is not clear, but genetic experiments are ongoing in our laboratory. Of the seven seedlings derived from 'Ol-ympia' × 'Yoho', two plants alone were sucrose accumula-tors, and the remaining five were hexose accumulators. In a population of 23 F2 population (self-seedlings of TSA-4), fifteen plants were fruited and all sucrose accumulators. These results suggest that the trait of sucrose accumulation may be controlled by recessive genes. In 2007, a total of 1111 seeds was obtained by self-pollination of TSA-4 for further genetic analysis of F2 population. A g r o / b i o c h e m i c a l   t r a i t s :  The Table shows comparison of agro/biochemical traits between tetraploid sucrose-accumulating grape genotype and its source culti-var. TSA-1 and TSA-4 reached full maturity in early Au-gust, and TSA-2 and TSA-3 in mid-August. Single berry weight ranged from 5.1 g (TSA-1) to 9.2 g (TSA-4) with a mean of 6.7 g. The berry weights of induced autotetra-ploids (TSA-1 to -3) were 1.3 to 1.5 times greater than those of source diploid cultivars, agreeing with a previous report (NOTSUKA et al. 2000). The allotetraploid TSA-4 is a more promising breeding material for production of large-sized berries with a maximum berry weight of 10.5 g. All genotypes had well-colored bunches, and skin color was blue-black, except for TSA-3 that was red. SSC ranged from 15.0 °Brix (TSA-3) to 18.4 °Brix (TSA-4) with an T a b l eComparison of agro/biochemical traits between tetraploid sucrose-accumulating grape genotype and its source cultivar (in bracket)Sucrose accumulatorTime of full maturity Berry weight (g)Skin colorSSC (°Brix)Acidity(%)Total sugar content(g·100 ml-1 juice)Sugar composition (%)Fru. Glu. Suc.TSA-1(Buffalo)Early Aug.Early Aug.5.14.0Blue-blackBlue-black18.119.00.500.5216.417.141.040.739.538.319.521.0TSA-2(Steuben)Mid. Aug.Late Aug.6.34.2Blue-blackBlue-black17.917.20.480.4616.215.242.038.836.838.221.223.0TSA-3(Red Port)Mid. Aug.Mid. Aug.5.94.0RedRed15.015.20.670.6313.313.939.745.341.542.418.812.2TSA-4(Olympia)(Yoho)Early Aug.Late Aug.Mid. Aug.9.212.08.2Blue-blackRed-brownRed18.418.017.00.500.450.5717.516.515.539.049.150.838.850.949.222.2tracetraceaverage of 17.4 °Brix. A similar trend was found in the total sugar content. Acidity ranged from 0.48 % (TSA-2) to 0.67 % (TSA-3) with an average of 0.54 %. On a percent-age basis, fructose and glucose presented in an approxi-mately 1:1 ratio. Sucrose varied from 18.8 % (TSA-3) to 22.2 % (TSA-4) with an average of 20.4 %. In their review of sugar composition, LOTT and BARRETT (1967) indicated that the sucrose content of nine diploid sucrose accumula-tors varied from 15.2 % (Bath) to 32.6 % (Sweet Blue), and averaged 25.5 %. Sucrose content of tetraploids was not always higher than that of diploids, suggesting that the effect of polyploidy is not clear for sucrose accumulation in grape berries. Progeny tests revealed that tetraploid su-crose-accumulating genotypes were essentially tetraploids in their breeding behavior. These genotypes with high su-crose content, which are early- to mid-season in maturity, are suitable for developing tetraploid table grapes. DAVIS, C.; ROBINSON, S. P.; 1996: Cloning of two putative vacuolar inver-tase cDNAs and their expression in grapevine tissues. Plant Physiol. 111, 275-283.LOTT, R. V.; BARRETT, H. C.; 1967: The dextrose, levulose, sucrose, and acid content of the juice from 39 grape clones. Vitis 6, 257-268.NOTSUKA, K.; TSURU, T.; SHIRAISHI, M.; 2000: Induced polyploid grapes via in vitro chromosome doubling. J. Japan. Soc. Hort. Sci. 69, 543-551.REISCH, B. I.; PRATT, C.; 1996: Grapes. In: J. JANICK and J. N. MOORE (Eds): Fruit breeding. Vine and small fruit crops, Vol. 2, 297-369. Wiley, New York, USA.SHIRAISHI, M.; 1993: Three descriptors for sugars to evaluate grape germ-plasm. Euphytica 71, 99-106.SHIRAISHI, M.; 2000: Comparison in changes in sugars, organic acids and amino acids during berry ripening of sucrose- and hexose-accumu-lating grape cultivars. J. Japan. Soc. Hort. Sci. 69, 141-148.TAKAYANAGI, T.; YOKOTSUKA, K.; 1997: Relationship between sucrose accumulation and sucrose-metabolizing enzymes in developing grapes. Am. J. Enol. Vitic. 48, 403-407.WINKLER, A. J.; COOK, J. A.; KLIEWER, W. M.; LIDER, L. A.; 1974: Develop-ment and composition of grapes. In: General viticulture: 138-196. Univ. California Press, Berkeley, Los Angeles, London.Received December 10, 2007

Tetraploid sucrose-accumulating grapevines

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Tetraploid sucrose-accumulating grapevines

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