A new and distinct variety of peach tree, which has been given the designation ‘V75024’, is a high quality mid-to-late season non-melting fleshed clingstone peach that produces high quality mid-to late season fruit suitable for processing as a canned or frozen product.

Patent
   PP16469
Priority
Jan 29 2001
Filed
Jan 10 2002
Issued
Apr 25 2006
Expiry
Jan 10 2022
Assg.orig
Entity
unknown
0
2
n/a
1. A new and distinct peach tree variety substantially as shown and described herein.

Peach cultivar ‘V75024’ originated at the Horticultural Research Institute of Ontario (now the Department of Plant Agriculture, University of Guelph) at Vineland Station, Ontario, Canada. It arose as a single selection from the seedling population that was derived from a controlled cross of ‘Suncling’ (patent status unknown) and ‘New Jersey Cling 81’ (patent status unknown) made in 1975. A large group of these first generation trees were planted and maintained on their own root system, during which time one seedling, which was the present variety, exhibited the outstanding desirable fruit characteristics described below and was selected in 1981.

The peach cultivar ‘V75024’ was asexually propagated since 1981 by budding on peach seedling rootstock in the research nursery at Vineland Station, Vineland, Ontario, Canada. Comparative records of performance of ‘V75024’ have been kept on its performance at this site since that date. The asexual propagation demonstrates that such reproduction of the characteristics of the tree are consistent and are established or transmitted through succeeding generations.

The new and distinct peach ‘V75024’ is a non-melting fleshed clingstone processing variety. The ‘V75024’ is self-pollinating and self-fruitful. It is a mid- to late-season maturing peach that produces high quality non-melting fleshed clingstone fruit suitable for processing as a canned or frozen product. It ripens 2 to 5 days later than ‘Babygold 5’ (patent status unknown), the variety it most closely resembles. Further compared to ‘Babygold 5’, ‘V75024’ has slightly shorter and narrower leaves, a flat leafblade profile, has resistance to bacterial spot, and larger fruits. DNA fingerprints of ‘V75024’ show a characteristic fragment length of 255 using SSR Pchcms2 and characteristic fragment lengths of 143, 142, 141, 140, 139, 138, and 136 using SSR UDP96-013 that are absent in ‘Babygold 5’.

The claimed variety differs from its parent, ‘Suncling’, in that: (1) Its ground skin color is cream-yellow (RHS 10) with dark red punctuation (RHS 480) coloring on approximately 75% of the fruit surface, whereas the skin color of ‘Suncling’ is golden with some blush when exposed to sun. (2) The stone of the claimed variety is a semi-freestone whereas the stone of ‘Suncling’ is a non-melting cling stone.

The accompanying photographic illustrations show typical specimens of the fruit and a set of DNA fingerprint profiles of the present new peach variety. The plants were grown in Vineland, Ontario.

FIG. 1. The illustration shows shoes the fruit of ‘V75024’ on the tree.

FIG. 2. The illustration shows an exterior and sectional view of a fruit divided in its suture plane to show flesh colour, pit cavity, and the stone remaining in place. The photographic illustration of the fruit was taken shortly after being picked and the colors are as true as is reasonably possible in a colour representation of this variety.

FIG. 3. Is a photograph of a gel with a series of five SSR DNA markers for ‘V75024’ compared to varieties ‘Babygold 5’, ‘Babygold 7’ and ‘Catherina’.

FIG. 4. Is a an overlay of the gel depicted in FIG. 3 with the bands detected by gel analysis.

The following detailed description of the new cultivar was obtained through the observation of plants growing at Vineland, Ontario. The observed plants were approximately 14 years of age, and were grown under typical soil and weather conditions for the area. Tree spacing was 12 feet within and 20 feet between rows. Color designations are with reference to The Royal Horticultural Society Color Chart. Common color terms are to be accorded their customary dictionary significance.

Although the new ‘V75024’ cultivar of peach tree possesses the above-described characteristics as a result of the growing conditions at Vineland, Ontario, it is to be expected that variations of the usual type and magnitude may appear that are caused by differences in growing conditions, fertilization, pruning, pest control and other horticultural practices when the new cultivar is grown in different environments.

Young leaf tissue samples of V75024, Babygold 5, Babygold 7, and Catherina were collected in May and September 2003 from nurseries located near Vineland, Ontario. The tissue was immediately frozen in liquid nitrogen, transported on dry ice and frozen at −80° C. DNA was extracted according to Serrano et al., 2002 with some modifications. Approximately 50 mg of leaf tissue were ground in a Fast Prep machine (QBioGene) with 200 uL extraction buffer (100 mM Tris-HCl, 1.4 M NaCl, 20 mM EDTA, 2% CTAB, 1% PVP, 0.2% β-mercaptoethanol, 0.1% NaHSO3) and incubated at 65° C. for 30 minutes. An equal volume of chloroform:isoamyl alcohol (24:1) was added, mixed by inversion and centrifuged at 6,000×g for 15 minutes. The supernatant was transferred to a new tube, mixed with 10 ug RNAse A and incubated at 37° C. for 30 minutes. An aliquot of 130 uL ice-cold isopropanol was added, mixed by inversion and centrifuged at 13,000×g for 5 minutes to recover the nucleic acid precipitate. The supernatant was removed and the pellet was washed with 400 uL of 10 mM ammonium acetate in 76% ethanol for 10 minutes and dried at room temperature. The pellet was re-suspended overnight at 4° C. in 200 uL of sterile ddH2O (ph 8.0) and quantified using a Spectramax Plus spectrophotometer (Molecular Devices). The DNA was then stored at −20° C.

Microsatellite primer pairs previously developed in peach (Table 1) were synthesized by Sigma Genosys (Oakville, ON). PCR was performed in a Robocycler 96 (Stratagene) and consisted of one incubation of 5 minutes at 94° C. followed by 35 cycles of 45 seconds at 94° C., 45 seconds at the appropriate annealing temperature (Table 1) and 1 minute at 72° C. The final cycle was followed by a 5 minute incubation at 72° C. Amplification reactions were carried out in 25 uL volumes containing 10 mM Tris-HCl pH 8.3, 50 mM KCl, 1.5 mM MgCl2, 100 uM of each dNTP, 10 pmol of each primer, 1U Taq polymerase (Sigma) and approximately 200 ng peach genomic DNA. Following amplification, 10 uL of the PCR reaction was separated on a 1% agarose gel containing ethidium bromide to verify PCR amplification. Samples were then stored at −20° C. prior to polyacrylamide gel electrophoresis.

TABLE 1
List of microsatellite markers (SSRs) used to
identify peach varieties
Annealing
SSR primer Sequence temperature
pair Sequence (5′-3′) ID (° C.) Reference
CPPCT030- TGAATATTGT- No. 1 50 Aranzana
A TCCTCAATTC et al.,
CPPCT030- CTCTAGGCAA- No. 2 (2002)
B GAGATGAGA
Pchcms2-A AGGGTCGTCT- No. 3 50 Sosinski
CTTTGAC et al.,
Pchcms2-B CTTCGTTTCA- No. 4 (2000)
AGGCCTG
Pchcms5-A CGCCCATGAC- No. 5 50 Sosinski
AAACTTA et al.,
Pchcms5-B GTCAAGAGGT- No. 6 (2000)
ACACCAG
UDP96-013- ATTCTTCACT- No. 7 57 Cipriani
A ACACGTGCAC- et al.,
G (1999),
UDP96-013- CCCCAGACAT- No. 8 Testolin
B ACTGTGGCTT et al.,
(2000)
UDP98-407- AGCGGCAGGC- No. 9 52 Cipriani
A TAAATATCAA et al.,
UDP98-407- AATCGCCGAT- No. 10 (1999),
B CAAAGCAAC Testolin
et al.,
(2000)
BPPCT025- TCCTGCGTAG- No. 11 57 Dirlewanger
A AAGAAGGTA- et al.,
GC (2002)
BPPCT025- CGACATAAAG- No. 12
B TCCAAATGGC
Pchgms1-A CGGTAAATAT- No. 13 52 Sosinski
GCCCATTGTG- et al.,
CAATC (2000)
Pchgms1-B GGATCATTGA- No. 14
ACTACGTCAA-
TCCTC

The PCR products were denatured by addition of an equal volume of sample buffer (95% de-ionized formamide, 10 mM NaOH, 0.05% bromophenol blue, 0.05% xylene cyanol). Samples were heated at 94° C. for 6 minutes and immediately placed on ice. Two microliters of each denatured preparation were loaded onto a 38×50 cm 6% polyacrylamide sequencing gel containing 7 M urea in 1× TBE buffer (90 mM Tris, 90 mM boric acid, 1 mM EDTA). Gels were run on a Sequi-Gen GT Nucleic Acid Electrophoresis Cell (Bio-Rad) for 4-5 hours at 85W. Following electrophoresis, gels were silver-stained using the SilverXpress Silver Staining Kit (Invitrogen), scanned on a HP scanjet 5470c scanner and dried on a Slab Gel Dryer SGD2000 (Savant). Fragment sizes were determined with 10-bp and 25-bp ladder DNA sizing markers (Invitrogen) using The Discovery Series, Quantity One software program (Bio-Rad).

The microsatellite primer pairs revealed genetic differences between and among the four peach varieties (FIGS. 1 and 2). Table 2 summarizes the fragment sizes detected for each of the primer pairs used based on repeated samples. There are a number of fragments that can be used to distinguish each of the four varieties using pairwise or any combination of comparisons. For example, fragments 262/264 from SSR BPPCT025 which are present in V75024, Catherina and Babygold 7 are not detected in Babygold 5. The characteristic fragment sizes that can be used to distinguish V75024 from all three reference varieties are presented in bold type in Table 2. These characteristic fragments are fragment 255 using SSR Pchcms2 and fragments 143, 142, 141, 140, 139, 138, 137, and 136 using SSR UDP96-013. These fragments are produced from DNA extracts of V75024 but are absent in Babygold 5, Babygold 7, and Catherina.

TABLE 2
Fragment sizes (bp) detected for four peach
varieties using seven microsatellite primer pairs.
Fragment sizes listed in bold are unique fragment sizes
detected in V75024 that are not detected in the other
three varieties.
Peach Variety
SSR V75024 Babygold 5 Babygold 7 Catherina
CPPCT030 200, 198, 196, 190, 188, 191, 190, 188, 200, 198,
195, 190, 189, 187, 184 184 197, 195,
187, 185 190, 189,
187, 185
Pchcms2 255, 245 245 246 246
Pchcms5 183, 181, 174, 183, 181, 183, 181, 174, 183, 181,
171, 169, 166, 174, 171, 171, 169, 166, 174, 171,
164 170, 166, 164 169, 166,
164 164
UDP96-013 143, 142, 141, 197, 196, 197, 195, 194, 197, 196,
140, 139, 138, 194, 192 191 194, 192
137, 136
UDP98-407 205, 204, 202, 204, 202, 207, 205, 203, 205, 203,
200, 190, 182, 200, 190, 190 202, 200,
181 182, 181 190
BPPCT025 304, 265, 263, 304, 281, 312, 263, 262, 305, 264,
204, 202, 200, 279, 204, 203, 201, 199, 262, 204,
198, 196, 194, 202, 200, 197, 195, 194 202, 200,
193 198, 197, 198, 196,
194 194, 192
Pchgms1 191, 190 192, 190 192, 190 192, 190
FIG. 1. Silver stained 6% polyacrylamide sequencing gel of four peach cultivars (1 = Babygold 5, 2 = V75024, 3 = Catherina, 4 = Babygold 7) using seven microsatellites (A = pchgms1, B = BPPCT025, C = UDP98-407, D = UDP96-013, E = pchcms5, F = pchcms2, G = CPPCT030).
FIG. 2. Silver stained 6% polyacrylamide sequencing gel of four peach cultivars (1 = Babygold 5, 2 = V75024, 3 = Catherina, 4 = Babygold 7) using seven microsatellites (A = pchgms1, B = BPPCT025, C = UDP98-407, D = UDP96-013, E = pchcms5, F = pchcms2, G = CPPCT030) showing banding patterns identified using Quantity One software.

Aranzana M.J., Garcia-Mas, J., Carbo, J. and P. Arus. 2002. Development and variability analysis of microsatellite markers in peach. Plant Breeding. 121:87-92.

Cipriani G., Lot G., Huang W-G., Marrazzo M.T., Peterlunger E. and R. Testolin. 1999.

AC/GT and AG/CT microsatellite repeats in peach [Prunus persica (L) Batsch]: isolation, characterization and cross-species amplification in Prunus. Theor. Appl. Genet. 99:65-72.

Dirlewanger E., Cosson P., Tavaud M., Aranzana M.J., Poizat C., Zanetto A., Arus P. and F. Laigret. 2002. Development of microsatellite markers in peach [Prunus persica (L.) Batsch] and their use in genetic diversity analysis in peach and sweet cherry (Prunus avium L.). Theor. Appl. Genet. 105:127-138.

Serrano B., Gomez-Aparisi J and J.I. Hormaza. 2002. Molecular fingerprinting of Prunus rootstocks using SSRs. J. Hort. Sci. Biotech. 77(3):368-372.

Sosinski B., Gannavarapu M., Hager L.D., Beck L.E., King G.J., Ryder C.D., Rajapakse S., Baird W.V., Ballard R.E. and A.G. Abbott. 2000. Characterization of microsatellite markers in peach [Prunus persica (L.) Batsch]. Theor. Appl. Genet. 101:421-428.

Testolin R., Marrazzo T., Cipriani G., Quarta R., Verde I., Dettori M.T., Pancaldi M. and S. Sansavini. 2000. Microsatellite DNA in peach (Prunus persica L. Batsch) and its use in fingerprinting and testing the genetic origin of cultivars. Genome. 43:512-520.

Miles, Neil

Patent Priority Assignee Title
Patent Priority Assignee Title
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