Maternal Lineages of the Cultivated Strawberry, Fragaria ×Ananassa
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HORTSCIENCE 44(6):1562–1565. 2009. Table 1. Country of origin and the detected chloroplast DNA haplotype of 75 accessions of Fragaria ·ananassa and seven accessions Maternal Lineages of the Cultivated of its wild relatives. Accessions Country Haplotype Strawberry, Fragaria ·ananassa, F. ·ananassa cultivars in North America Allstar USA V Arking USA V Revealed by Chloroplast Blakemore USA V Cardinal USA V DNA Variation Chandler USA V Columbia USA C 1 Masanori Honjo , Sono Kataoka, Susumu Yui, and Masami Morishita Donner USA V National Agricultural Research Center for Tohoku Region, Nabeyashiki, 92 Douglas USA V Excelsior USA V Shimokuriyagawa, Morioka, Iwate 020-0123, Japan Fairfax USA V Fresno USA V Miyuki Kunihisa Geneva USA V National Institute of Vegetable and Tea Science, 3-1-1 Kannondai, Tsukuba, Hecker USA V Ibaraki 305-8666, Japan Holiday USA V Honeoye USA V Takayoshi Yano, Megumi Hamano, and Hiromichi Yamazaki Huxley USA V Lassen USA V National Agricultural Research Center for Tohoku Region, Nabeyashiki, 92 Linn USA V Shimokuriyagawa, Morioka, Iwate 020-0123, Japan Marshall USA V Missionary USA V Additional index words. Fragaria virginiana, Fragaria chiloensis, haplotype, maternal Pajaro USA V inheritance, maternal origin Premier (Howard 17) USA V Raritan USA V Abstract. We analyzed sequence variation in chloroplast DNA (cpDNA) to investigate the Selva USA V origin of the cultivated strawberry, Fragaria ·ananassa. From analysis of two noncoding Sequoia USA V regions, trnL–trnF and trnR–rrn5, we found three haplotypes (V, C, and X) in F. Tioga USA V ·ananassa. Haplotype V corresponded to the haplotype of F. virginiana and was Tyee Canada V possessed by cultivars bred over a wide geographic range, including North America, Vibrant Canada V Europe, and Japan. Almost all the North American cultivars analyzed in this study Wiltguard USA V possessed haplotype V, suggesting a founder effect. Haplotype C corresponded to the F. ·ananassa cultivars in Europe Cambridge Favourite UK X haplotype of F. chiloensis and was detected mainly in Japanese cultivars. Haplotype X Cambridge Prizewinner UK V was found in only two English cultivars. This haplotype was positioned as intermediate Deutch Evern Germany V between haplotypes V and C in a median-joining network and was considered to be Merton Princess UK X representative of the process of differentiation between F. virginiana and F. chiloensis. Oranda Netherlands V Results of controlled crosses indicate that cpDNA haplotypes of F. ·ananassa are Redgauntlet UK V maternally inherited. These results verify that F. ·ananassa is an interspecific hybrid Senga Sengana Germany V between F. virginiana and F. chiloensis and indicate that traditional cultivars of F. F. ·ananassa cultivars in Japan ·ananassa have been derived from at least three maternal lineages. We demonstrate that Aiberry Japan V the cpDNA variation detected in this study can be used to verify parentage and for Akihime Japan V Asuka Wave Japan C extending hypotheses about June yellows, a leaf variegation disorder in strawberry. Bellerouge Japan V Benihoppe Japan V Decorouge Japan V Enrai Japan V Determining the historical pedigree of in that first hybridization was claimed to have Everberry Japan V cultivars is an important step in understand- been only five plants, which were introduced Fukuba Japan C ing the evolution of crop species (Matsuoka, from Chile to Europe in 1714 (Darrow, 1966). Harunoka Japan V 2005). It can also help to avoid inbreeding Systematic breeding began in England in the Haruyoi Japan V depression and to identify the origin of early 1800s and in North America in the mid- Hatsukuni Japan C agronomically important traits. 1800s using a small number of native and Himiko Japan C The cultivated strawberry, Fragaria cultivated clones (Darrow, 1966). Most mod- Hokowase Japan V ·ananassa Duch., is one of the most eco- ern strawberry cultivars are the progeny of this Kitanokagayaki Japan V nomically important fruit crops in the world. relatively narrow range of germplasm, al- Kogyoku Japan V Kurume 34 Japan C It first arose from accidental hybridization though attempts have been made recently to Kurume 103 Japan V between two American octoploid species, F. increase genetic diversity by using wild ge- Miyazaki Japan V virginiana and F. chiloensis, in a European netic resources (Hancock et al., 2001; Luby Mo-ikko Japan V garden during the early to mid-1700s (Darrow, et al., 2008). On the basis of pedigree data, Morioka 16 Japan V 1966). The number of F. chiloensis involved Dale and Sjulin (1990) reported that the Morioka 26 Japan V majority of modern North American cultivars Morioka 30 Japan V came from only 17 cytoplasmic sources. Morioka 32 Japan V However, further tracing was impossible ow- Morioka 33 Japan V Received for publication 19 June 2009. Accepted ing to incomplete records. Morioka 34 Japan V for publication 12 Aug. 2009. Natsuakari Japan V Because chloroplast DNA (cpDNA) is Nyoho Japan V We thank Naoe Suzuki for technical assistance; unaffected by changes in ploidy, which can Rikiya Kimura, Setsuko Oki, Yukari Sakurai, and Ohishi Shikinari Japan V Keiko Iwabuchi for help in cultivation of plant complicate phylogenetic analysis, the ge- Reiko Japan V materials; and anonymous reviewers for their valu- nome is particularly useful for the phyloge- Sachinoka Japan C able comments. netic analysis of Fragaria. Potter et al. (2000) Sagahonoka Japan C 1To whom reprint requests should be addressed; examined the sequence variation in cpDNA e-mail [email protected]. among 14 species of Fragaria with various (Continued on next page) 1562 HORTSCIENCE VOL. 44(6) OCTOBER 2009 Table 1. (Continued) Country of origin and the Table 2. Chloroplast DNA primers used in the analysis of Fragaria ·ananassa, annealing temperature, and detected chloroplast DNA haplotype of 75 references. accessions of Fragaria ·ananassa and seven Annealing accessions of its wild relatives. Primer Sequence 5#-3# temp (°C) References Accessions Country Haplotype trnL (UAA) GGTTCAAGTCCCTCTATCCC 50 Taberlet et al. (1991) Summer Candy Japan C trnF (GAA) ATTTGAACTGGTGACACGAG Taberlet et al. (1991) Summer Princess Japan V trnR (ACG) CGACACCGTGGTTCGTAGC 50 Yoshimura et al. (unpublished data) Summerberry Japan V rrn5 TGGTGTCCCAGGCGTAGAG Yoshimura et al. (unpublished data) Tochihitomi Japan C Tochiotome Japan C Toyonoka Japan C MgCl2, 0.9 units Taq polymerase, 0.2 mMof Table 3. Chloroplast DNA haplotype of seedlings Yachiyo Japan V each primer, and 10 ng template DNA in obtained from controlled cross-pollination of Wild relatives a total volume of 30 mL. Thermocycling cultivars of Fragaria ·ananassa. F. virginianaz V y conditions were as follows: 3 min at 94 °C; Parentage (female · male) Haplotype F. virginiana V2 30 cycles of 30 s at 94 °C, 45 s at 50 °C, and F. chiloensis C Haplotype C · haplotype V ‘PI551445’ 45 s at 72 °C; and a final extension step at Sagahonoka · Natsuakari, No.1 C F. vesca EMC Vesca 72 °C for 5 min. The PCR was carried out in Sagahonoka · Natsuakari, No.2 C F. vesca UC1 Vesca a GeneAmp PCR System Model 9700 (Ap- Sagahonoka · Natsuakari, No.3 C F. vesca UC5 Vesca plied Biosystems, Foster City, CA) or a PCR Sagahonoka · Natsuakari, No.4 C F. nilgerrensisz Nil Thermal Cycler Dice (Takara, Tokyo, Japan). Sagahonoka · Natsuakari, No.5 C z Sagahonoka · Selva, No.1 C Germplasm conserved in the National The PCR products were purified with a QIA- Sagahonoka · Selva, No.2 C Agricultural Research Center for Tohoku region, quick PCR purification Kit (Qiagen GmbH, Haplotype V · haplotype C Morioka, Japan. Hilden, Germany). The DNA obtained was y Natsuakari · Sagahonoka, No.1 V Seeds were purchased from the B&T World sequenced with an ABI PRISM 310 Genetic Natsuakari · Sagahonoka, No.2 V Seeds, Paguignan, France, in 2002. Analyzer (Applied Biosystems) using a Big- Natsuakari · Sagahonoka, No.3 V Dye Terminator Cycle Sequencing Ready Selva · Sagahonoka V Reaction Kit (Applied Biosystems). Se- Haplotype V · haplotype X ploidy levels. Distinctive sequences for each quencing data were aligned manually with (Morioka 32 · Sagahonoka) species, including F. virginiana and F. chi- CLUSTAL W (Thompson et al., 1994). In- · Cambridge Favourite V loensis, were found in the trnL–trnF region, Haplotype X · haplotype V sertions/deletions (indels) were generally and F. ·ananassa showed the same sequence Cambridge Favourite · Aiberry X placed so as to increase the number of as F. virginiana. Although this analysis Cambridge Favourite · X matching nucleotides in a sequence position. examined only one sample of F. ·ananassa Kitanokagayaki We determined cpDNA haplotypes from and therefore did not provide further infor- Cambridge Favourite x Morioka 32 X nucleotide substitutions and indels. To show Haplotype X · haplotype C mation about the involvement of other wild the relatedness of haplotypes, we constructed Cambridge Favourite · Sachinoka X species in the establishment of F. ·ananassa, a median-joining network (Bandelt et al., sequence variation in cpDNA may allow us 1999) with epsilon value set to zero using to infer the origin of each strawberry cultivar. the software Network 4.5.0.0 (http://www. Japanese cultivars and the American cultivar In this study, we investigated the mode fluxus-engineering.com). Columbia. We detected haplotype X in only of inheritance of cpDNA and then identified To confirm the mode of inheritance of two English cultivars, Cambridge Favourite the maternal origin of F. ·ananassa. We dis- cpDNA in F. ·ananassa, we performed and Merton Princess. cuss the accuracy of the reported pedigree of controlled cross-pollination between culti- All seedlings obtained from controlled cultivars and the relationship between vars possessing different haplotypes (Table cross-pollination possessed the same haplo- cpDNA haplotype and June yellows, a leaf 3). We examined the haplotype for one to five types as their maternal parents (Table 3). variegation disorder in strawberry. seedlings per cross. Discussion Materials and Methods Results The results of the controlled crosses We collected fresh leaves from 75 acces- We found two haplotypes (V and V2) in (Table 3) indicate that the cpDNA of F.