Neo Muscat Research Articles

Virus-Induced Flowering by Apple Latent Spherical Virus Vector: Effective Use to Accelerate Breeding of Grapevine.

Apple latent spherical virus (ALSV) was successfully used in promoting flowering (virus-induced flowering, VIF) in apple and pear seedlings. In this paper, we report the use of ALSV vectors for VIF in seedlings and in vitro cultures of grapevine. After adjusting experimental conditions for biolistic inoculation of virus RNA, ALSV efficiently infected not only progeny seedlings of Vitis spp. ‘Koshu,’ but also in vitro cultures of V. vinifera ‘Neo Muscat’ without inducing viral symptoms. The grapevine seedlings and in vitro cultures inoculated with an ALSV vector expressing the ‘florigen’ gene (Arabidopsis Flowering locus T, AtFT) started to set floral buds 20–30 days after inoculation. This VIF technology was successfully used to promote flowering and produce grapes with viable seeds in in vitro cultures of F1 hybrids from crosses between V. ficifolia and V. vinifera and made it possible to analyze the quality of fruits within a year after germination. High-temperature (37 °C) treatment of ALSV-infected grapevine disabled virus movement to newly growing tissue to obtain ALSV-free shoots. Thus, the VIF using ALSV vectors can be used to shorten the generation time of grapevine seedlings and accelerate breeding of grapevines with desired traits.

CRISPR/Cas9-mediated targeted mutagenesis in grape.

RNA-guided genome editing using the CRISPR/Cas9 CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) system has been applied successfully in several plant species. However, to date, there are few reports on the use of any of the current genome editing approaches in grape—an important fruit crop with a large market not only for table grapes but also for wine. Here, we report successful targeted mutagenesis in grape (Vitis vinifera L., cv. Neo Muscat) using the CRISPR/Cas9 system. When a Cas9 expression construct was transformed to embryonic calli along with a synthetic sgRNA expression construct targeting the Vitis vinifera phytoene desaturase (VvPDS) gene, regenerated plants with albino leaves were obtained. DNA sequencing confirmed that the VvPDS gene was mutated at the target site in regenerated grape plants. Interestingly, the ratio of mutated cells was higher in lower, older, leaves compared to that in newly appearing upper leaves. This result might suggest either that the proportion of targeted mutagenized cells is higher in older leaves due to the repeated induction of DNA double strand breaks (DSBs), or that the efficiency of precise DSBs repair in cells of old grape leaves is decreased.

Breeding, genetics, and genomics of fruit trees.

Breeding, genetics, and genomics of fruit trees.

Advances in table grape breeding in Japan.

In Japan, few grape cultivars related to Vitis vinifera existed 200 years ago, on account of Japan’s high rainfall. Many V. labruscana and vinifera cultivars were introduced to Japan in the 19th century. Labruscana was grown instead of vinifera, mainly because of severe disease problems and a high incidence of berry cracking. Grape breeding for table use started in the 20th century, with the goal of combining the berry quality of vinifera with the ease of cultivation of labruscana. By 1945, three strategies were used: 1) crossing among introduced diploid vinifera and vinifera-related cultivars of Japanese origin, 2) interspecific crossing in tetraploid cultivars, and 3) interspecific crossing in diploid cultivars, resulting in ‘Neo Muscat’, ‘Kyoho’, and ‘Muscat Bailey A’. Later, tetraploid interspecific crossing over generations developed many ‘Kyoho’-related cultivars, including ‘Pione’, many of which have large berries, intermediate flesh texture between the two species, a labruscan or neutral flavor, and moderate disease resistance. Interspecific diploid crossing over generations developed ‘Shine Muscat’ in 2006, with large berries, crispy flesh, a muscat flavor, no cracking, seedless fruit by gibberellin application, and moderate resistance to downy mildew and ripe rot.

EVALUATION OF 17 TABLE GRAPE ACCESSIONS FOR FOLIAR RESISTANCE TO DOWNY MILDEW

Grapevine downy mildew (DM) could cause serious damage to vines and young clusters in a humid climate, and multiple fungicides are applied to control this disease. Since most grapes are produced for fresh fruit consumption in Japan, we evaluated foliar DM resistance of 17 table grape cultivars/selections that are important for Japanese table grape breeding by visual rating. By analysis of variance (ANOVA), we estimated the variance component of blocks, cultivars, years and the interaction between cultivars and years. ANOVA revealed that the effect of cultivars is the main source of the variance (79.6%). The effect of year was significant, whereas the estimated variance component was only 3.0% of total variance. The effect of blocks and the interaction were not significant. These results suggested that our evaluation of the DM resistance was reliable. Two Vitis vinifera cultivars ?Rizamat? and ?Neo Muscat? were highly susceptible, and were easily defoliated comparing to other accessions. Most of the other V. vinifera cultivars were susceptible, whereas ?Muscat Hamburg? showed some resistance to this disease. In contrast, some American hybrids such as ?Concord?, ?Campbell Early? and ?Ontario? were highly resistant. However, one of the American hybrids, ?Steuben?, was not as resistant as those resistant hybrids. Japanese tetraploid cultivars, ?Kyoho?, ?Pione? and ?Darkridge?, are rated as medium, while ?Ishiharawase?, which is the tetraploid bud sport of ?Campbell Early?, was highly resistant. These results suggest that there is a variation of DM resistance among table grape cultivars, and that the resistance of Japanese leading cultivars ?Kyoho? and ?Pione? are medium.

Development of SCAR Markers Linked to Cold Resistance for Marker-assisted Selection of Grapevines

포도 신품종 육성에서 내한성 계통의 조기 선발을 위하여, 다양한 품종을 대상으로 randomly amplified polymorphic DNA(RAPD)를 수행하고, 그 결과를 바탕으로‘Campbell Early’x ‘Kaiji’, ‘Golden Muscat’x‘Tamnara’,‘Campbell Early’x‘Neo Muscat’와‘Alden’x‘Kaiji’의 포도교배조합 실생계통을 이용하여 내한성 관련 sequence characterized amplified region(SCAR) 분자표지를 개발하였다. 400여개의 primer를 사용하여 bulk집단에서의 RAPD 분석을 통해서 6개의 UBC random primer를 최종적으로 선발하였고, RAPD 산물을 클로닝한 후 염기서열을 분석하였다. 6개의 SCAR primer (UBC123-428, UBC196-353, UBC221-503, UBC317-800, UBC342-500, UBC344-451)를 제작하고 교배조합 실생에 재검정한 결과 내한성과 관련한 특이밴드를 확인할 수 있었다. 포도나무의 내한성과 관련한 특이적인 밴드출현양상은 수피의 탄수화물함량, 전해질전도도, 포장에서의 신초생존율과 유사한 양상을 보였다. 본 연구에서 개발한 SCAR marker는 포도의 염색체상에서 위치를 확인할 수 있었으며, 그 주변에는 주로 cytochrome P450, glutathione S-transferase, leucine-rich repeat, pleiotropic drug resistance 12, ribosomal protein 등의 유전자가 분포하였다. 본 연구에서 개발된 포도 내한성 관련 SCAR marker는 내한성 포도 품종의 조기선발과 육종효율 증진에 기여할 수 있을 것으로 기대된다.

Comparison of the Antioxidative Effects and Content of Anthocyanin and Phenolic Compounds in Different Varieties of Vitis vinifera Ethanol Extract

This study was a quantitative HPLC analysis of four anthocyanins and five phenolic compounds contained in the skins, vines and seeds of the Campbell Early, Muscat Bailey A and Neo Muscat grape varieties. In the phenolic analysis, the seeds of the Campbell Early were found to contain 1.9, 1.8 and 1.6 times higher quantities of gallic acid, catechin and epicatechin relative to other grape seeds. Three anthocyanins, cyanidin, peonidin and pelargonidin, were also found to be higher in the skins of the Campbell Early relative to other grape skins. Therefore, the Campbell Early is the most useful grape variety with regard to the extraction of these six compounds from these grape seeds and skins. The free radical scavenging effects of grape seeds were also compared, and the results indicated that the Campbell Early seeds were most effective among them.

Inhibitory Effects of Neo Muscat Grape Vine Extracts on Melanin Biosynthesis

Neo Muscat grape vine extracts significantly inhibited melanin production in melanocyte stimulating hormone-stimulated B16 cells. They also exhibited tyrosinase inhibitory activity and reduced the levels of intra-cellular tyrosinase. Moreover, when the extracts were treated with UV B-induced hyper-pigmented brown guinea pig skin, depigmenting activity was observed, but with no visible edema. These results indicate that the vine extracts of Neo Muscat grapes might prove useful as a skin depigmenting agent.

Similarity of Amino Acid Composition of Grape Berries Produced in Different Years and Locations.

Free amino acids in the juice of four table grape cultivars ('Campbell Early', 'Kyoho', 'Muscat Bailey A', and 'Neo Muscat') grown in eight locations, primarily in the southwest part of Japan, were analyzed in 1992 and 1993. In each cultivar, total amino acids content ranged between 1.0 to 1.8 mM. To compare the similarity in the amino acid composition of berries produced in different years and locations, the pattern similarity index (PSI) was calculated. The PSI is defined as a cosine of the angle (θ) between the amino acid composition of environment A for vector OA (aAsp, aThr, …, aArg=a1, a2, …, an) and that of environment B for vector OB (bAsp, bThr, …, bArg=b1, b2, …, bn) in n dimensional space : [numerical formula] With regard to the amino acid composition (pattern), the PSI value consisted of the intuitive judgment of similarity on the pattern. As the number of significantly differentiated amino acids increased, the PSI correspondingly tended to be lower. Based on the PSI value, the criterion of classification for pattern similarity of amino acid composition was proposed as follows : over 0.980 as more ; below 0.979 as less similar. The percentage frequencies of more patterns, with respect to PSI values, 'Campbell Early', 'Muscat Bailey A', 'Neo Muscat', and 'Kyoho' were 75, 73, 72, and 69%, accounting for about 70% of all pairwise comparisons (nC2). They suggest that the amino acid composition in grape berries is highly similar even if the years and/or locations are different. Each pattern of amino acid composition in the cultivars examined are visually different from each other. When mean concentrations of the individual amino acid were calcuated by averaging two years in Fukuoka, the PSI values were 0.959 in 'Kyoho', 0.899 in 'Muscat Bailey A', and 0.753 in 'Neo Muscat' in contrast with 'Campbell Early'. 'Kyoho' vs. 'Muscat Bailey A', 'Kyoho' vs. 'Neo Muscat' and 'Muscat Bailey A' vs. 'Neo Muscat' gave less patterns with the PSI vaules of 0.979, 0.882, and 0.933, respectively. These results indicate that the amino acid composition in grape berries is a useful marker for ampelographic work.

Correlations between Specific Gravity and Soluble Solids Concentration in Grape Berries

We investigated correlations between specific gravity and soluble solids concentration in grape berries, since the nondestructive measurement of soluble solids concentration by near infrared spectroscopy is not put to practical use. Specific gravity of each cluster was measured by using an electronic balance and an acoustic volume meter that can measure volume without dipping fruits in water. Soluble solids concentration (Brix) of all berries in each cluster was measured using a refractometer. The relationship between specific gravity and Brix value of clusters of 'Kyoho' grape was constant and a linear one, regardless of the harvest year and the production district. The relation had a high correlation (r=0.981***) within 16-23° Brix with a standard error of 0.35° Brix. High correlations were also found between specific gravity and Brix value in clusters of 'Campbell Early', 'Neo Muscat' and 'Koshu' grape. The slope of the regression line varied little among the four varieties. These results suggest that the measurement of specific gravity with an acoustic volume meter is a useful method to measure nondestructively the soluble solids concentration of clusters.

Antibiotik sebagai Induktor Buah Tanpa Biji pada Anggur

Four kinds of antibiotics were examined, viz. streptomycin (SM) and gentamicin (GM) as bactericidal antibiotica and tetracycline (TC) and spectinomycin (SE) as bacteriostatical antibiotics. Kyoho an Pione (tetraploid hybris cultivars) were used as the plant materials. Among the four antibiotics, SM and SE were effective to induce seedlessness in Muscat of Alexandria (MOA) and less effective for Kyoho and Pione. The seedless induction effect of SM and SE seemed not to be affected by their mode of action. Using the four cultivars, Kyoho, Pione, MOA, and Neo Muscat (NM), either SM or SE most effective to induce seedlessness when applied 3 days before full bloom. The ineffectiveness of antibiotics for Kyoho and Pione was indicated by the higher percentage of empty-seeded berries in these cultivars compared to MOA and Nm. Since there was no normal seed in the antibiotic-treated berries, this phenomenon indicated that the outer parts of the tetraploid ovules were more resistant to antibiotic treatments than those of diploid ovules. Key word: Grape, Seedless, Antibiotics

Jasmonic acid effects on gibberellic acid-induced seedlessness in ‘Neo Muscat’ table grapes

SummaryThe effect of jasmonic acid (JA), applied alone or in combination with gibberellin A3 (GA3), on the induction of seedlessness in ‘Neo Muscat’ (Vitis vinifera) was investigated. Endogenous JA levels in the florets of ‘Neo Muscat’ and ‘Delaware’ (Vitis vinifera x Vitis labrusca) were also studied. The proportion of seedless berries induced by application of GA3 alone 12 d before full bloom (DBB) was 17.0%, and at 6 DBB it was 62.7%. The proportion of seedless berries induced by GA3 + JA applied 6 DBB increased significantly with increasing JA concentration, while pollen germination decreased quadratically with increasing JA concentration. JA did not affect seed number per berry or the seed’s fresh weight. Treatment with JA alone also induced seedlessness effectively. Although the fresh weight of the seedless berries induced by the application of GA3 + JA 12 DBB decreased quadratically with increasing JA concentration, there were no significant differences between the treatments at 6 DBB. Fruit soluble solids and titratable acidity were not affected by JA. There were no obvious differences in the amounts of JA in the florets of ‘Delaware’ and ‘Neo Muscat’.

Effects of Lateral Shoot Cutting on Population Density of the Chillie Thrips, Scirtothrips dorsalis HOOD(Thysanoptera:Thripidae)on Grape

Effects of lateral shoot cutting on population density of the chillie thrips, Scirtothrips dorsalis HOOD, on grape, cultivar Neo Muscat, were investigated in Okayama in 1990, Two experimental plots were designed, with cut or uncut lateral shoots. The numbers of adults and larvae collected by washing were much smaller in the cut plot than in the uncut plot, suggesting that their occurrence on shoots was influenced by the quantity of available resources (lateral shoots). On the other hand, the number of catches with yellow sticky traps was much greater in the cut plot than in the uncut plot. Results suggested that the number of catches reflected the frequency of adult dispersal and that the dispersal was more frequent in the cut plot because of the resource shortage. The number of the trap catches reflected the population fluctuation in each of the two plots, although the regression coefficient (X: the number of adults collected by washing method, Y: catches by yellow sticky trap) was extremely different between the two plots. Degree of damage to the fruit cluster did not differ between the two plots.

Breaking Bud Dormancy in Grapevines with Garlic Paste

Four dormant grapevine (Vitis spp.) cultivars grown under forced conditions were treated immediately after pruning with a paste of fresh garlic, the supernatant of a 20% suspension of CaCN2, and 50% `Merit' solution a foliar fertilizer, to break bud dormancy. Garlic paste significantly accelerated budbreak and increased the rate of budbreak in three cultivars, but for `Delaware,' CaCN2 was more effective. Garlic also affected budbreak of cuttings with a single bud of `Kyoho,' `Neo Muscat,' or `Muscat Bailey A' grapes in a deep stage of dormancy. Garlic paste applied to cool `Muscat of Alexandria' vines immediately after they were pruned in various stages of dormancy (from November to March) accelerated budbreak in the spring when application was made by January. The rate of budbreak was steady, but earlier CaCN2 treatments resulted in a rate of budbreak that was not uniform, although the first budbreak was accelerated. Low concentrations of garlic juice did not promote budbreak of cuttings of `Muscat of Alexandria'. When garlic paste was applied to various parts of cuttings with two buds, budbreak was accelerated when the upper cross-section was treated, but the overall rate of budbreak was highest when the lower half of the cutting was treated. Chemical name used: calcium cyanamide (CaCN2).

Cellular Variation in Chromosome Number Induced by p-Fluorophenylalanine in Grape Seedlings1

Abstract p-Fluorophenylalanine reduced germination and growth of diploid seedlings of grape (Vitis vinifera L. cv. Neo Muscat) and induced variation in chromosome numbers from haploidy to tetraploidy including aneuploidy in root cells.

Sensitivity of grapevines to injury by atmospheric sulfur dioxide

Sensitivity of grapevines to injury by sulfur dioxide were studied. One or two-years old plants were exposed to different concentrations of the gas (0.26, 0.13, 0.065, 0ppm) at different season in a green house. The results obtained are summarized as follows:1. Sulfur dioxide injury emerged on the middle leaves of grapevine at first, however the degree of leaf lesion was remarkable on the lowest leaves. Leaf lesion and early leaf fall increased in proportion to the gas concentrations.2. Six varieties of grapevine used varied considerably in their sensitivity to the gas. Variety of Fredonia was highly sensitive and Delaware grape was fairly sensitive. Kyoho and Neo muscat grapes were intermediate. Kyogei grape was resistant, Koshu grape was the most resistant and was not suffered from leaf injury through exposure periods. Fredonia grape was absorbed the gas more than Koshu grape.3. Sensitivity of Fredonia grape varied with season (stage) and increased from spring to autumn. Absorption of the gas also increased.4. Fredonia and Delaware grapes were exposed to the gas from May 23 to Oct. 29 of 1967 to clarify the effect of preceding year exposure on shoot growth and cluster formation. Degree of leaf lesion of Fredonia grape became remarkable in proportion to the gas concentrations and the extention of exposure duration. Also leaf lesion of Delaware grape formed abundantly at the concentration of 0.26ppm after extension of the exposure duration. After terminating the gas exposure, new shoot growth and cluster formation of Fredonia grape decreased in 1968 with increasing gas concentrations, corresponding the degree of leaf lesion and falling in 1967. Shoot growth of Delaware grape decreased at 0.26ppm and cluster formation also decreased at 0.13ppm and above.