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Akiko Watari, Toshio Hanada, Hisayo Yamane, Tomoya Esumi, Ryutaro Tao, Hideaki Yaegaki, Masami Yamaguchi, Kenji Beppu and Ikuo Kataoka

. ( Romero et al., 2004 )]. Recently, pollen S component SFB ( S haplotype-specific F-box protein gene) has also been characterized in almond ( Ushijima et al., 2003 ), sweet and sour cherries ( Yamane et al., 2003b ), japanese apricot ( Entani et al

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Axel O. Ramírez-Madera and Michael J. Havey

cucumbers, and revealed that in spite of different haplotypes, all three sources of ZYMV resistance encode the same protein sequence. Materials and Methods DNA was isolated from cucumber accessions ZYMV-resistant TMG-1 ( Park et al., 2000 ; Provvidenti

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Toshio Hanada, Kyoko Fukuta, Hisayo Yamane, Tomoya Esumi, Ryutaro Tao, Thomas M. Gradziel, Abhaya M. Dandekar, Ángel Fernández i Martí, José M. Alonso and Rafel Socias i Company

et al., 2003 ) for the pistil and pollen specificities, respectively. Because the pistil and pollen determinant genes have been identified, the variants of the S-RNase and SFB combinations are called S haplotypes and the variants of a given S

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Su-Hyoung Park, Ki-Taek Kim, Sun-Hyoung Lim, Moo-Kyoung Yoon, Soo-Seong Lee, Changhoo Chun and Hyo-Geun Park

Self-incompatibility (SI) in Brassicaceae vegetables prevents self-pollination by recognizing self-pollens and rejecting them at the stigmatic surfaces. The S-haplotypes of 47 hybrid radish cultivars that are commercially available in Korea were classified and identified using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Twelve kinds of S-haplotypes were identified from the cultivars: S 1 , S 8 , S 11 , S 17 , S 18 , S 30, and S 31 haplotypes in class-I S-haplotype and S 4 , S 5 , S 13 , S 21, and S 26 haplotypes in class-II S-haplotypes. Even though the class-II S-haplotypes are supposed to exhibit weak and/or leaky SI activity, the class-II S-haplotypes showed the same allele frequency of class-I S-haplotypes in 38 fully classified commercial cultivars. The SI activity was examined using the pollen tube germination test, flower pollination test, and the seed set ratio analysis. The pollen tube test showed low correlation (R 2 = 0.13) with the flower pollination test, a conventional method. The results of seed set ratio analysis varied from 0% to 159%, and thus could distinguish the weak and strong SI activity clearly and showed high correlation with the flower pollination test (R 2 = 0.69). The seed set ratios of the cultivars possessing the class-I/class-I, class-I/class-II, and class-II/class-II genotypes were 0.6%, 17.4%, and 38.1%, respectively. Among the eight class-II/class-II cultivars, three cultivars showed strong SI activity. The SI activity of the S 4 S 17 , S 5 S 8, and S 4 S 26 genotypes varied among cultivars, but the S 1 S 17 , S 5 S 17, and S 8 S 26 genotypes showed constant strong, intermediate, and strong activity, respectively, among the cultivars. Results indicate that the SI activity of Brassicaceae vegetables depends not only on the S-haplotypes, but also on the genetic background of cultivars.

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Attila Hegedüs, Zoltán Szabó, József Nyéki, Júlia Halász and Andrzej Pedryc

The most commercially grown peach [Prunus persica (L.) Batsch.] cultivars do not require cross-pollination for reasonable fruit set; however, self-incompatibility is a well-known feature within the Prunoideae subfamily. Isoelectric focusing and native polyacrylamide gel electrophoresis of S-ribonucleases; PCR analyses of S-RNase and S-haplotype-specific F-box genes as well as DNA sequencing were carried out to survey the self-(in)compatibility allele pool and to uncover the nature of self-compatibility in peach. From 25 cultivars and hybrids with considerable diversity in phenotype and origin, only two S-haplotypes were detected. Allele identity could be checked by exact length determination of the PCR-amplified fragments and/or partial sequencing of the peach S 1-, S 2-, and Prunus davidiana (Carr.) Franch. S 1-RNases. S-RNases of peach were detected to possess ribonuclease activity, and a single nucleotide polymorphism in the S 1-RNase was shown, which represents a synonymous substitution and does not change the amino acid present at the position in the protein. A 700-bp fragment of the peach SFB gene was PCR-amplified, which is similar to the fragment size of functional Prunus L. SFBs. All data obtained in this study may support the contribution of genes outside the S-locus to the self-compatible phenotype of peaches.

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K. Ikeda, A. Watari, K. Ushijima, H. Yamane, N.R. Hauck, A.F. Iezzoni and R. Tao

S4′ is a pollen-part mutant in sweet cherry (Prunus avium L.) that is extensively used to develop self-compatible cultivars. The S4′-haplotype is known to have a functional stylar component and a nonfunctional pollen component. The pollen component in sweet cherry necessary for the specificity of the pollen reaction is believed to be an S-haplotype specific F-box protein gene, called SFB. This study describes two molecular markers that distinguish between SFB4 and SFB4′ by taking advantage of a four base pair deletion in the mutant allele. The resulting polymerase chain reaction (PCR) products can either be separated directly on a polyacrylamide gel or they can be subjected to restriction enzyme digestion and the different sized products can be visualized on an agarose gel. The latter technique utilizes restriction sites created in the PCR products from the SFB4′ allele, but not the SFB4 allele. Because the primer sets created differential restriction sites, these primer sets were termed dCAPS (derived cleaved amplified polymorphism sequence) markers. These molecular assays can be used to verify self-compatibility conferred by the S4′-haplotype.

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Christopher M. Long, Colleen A. Mulinix and Amy F. Iezzoni

Microspore-derived callus cultures were obtained by anther culture of `Emperor Francis' sweet cherry (Prunus avium L.). Branches were removed from the field in January and March and forced in the laboratory. When the microspores reached the uninucleate stage, anthers were placed on modified Quoirin and Lepoivre liquid culture medium containing 4.4 μm BA and 4.5 μm 2,4-D. After ≈60 days, callus that emerged from the anthers was placed on woody plant medium supplemented with 1 μm 2,4-D and 3 μm 2iP and routinely transferred. The resulting 270 callus cultures were screened for two allozymes heterozygous in `Emperor Francis', Pgi-2 and 6-Pgd-1. Of the 270 callus cultures, 154 expressed only one allele each for Pgi-2 and 6-Pgd-1; thus, they were considered microspore-derived. The microspore-derived callus cultures can be used as a linkage mapping population. Chemical names used: 6-benzyladenine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); N6-(2-isopentenyl)-adenine (2iP).

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Masanori Honjo, Sono Kataoka, Susumu Yui, Masami Morishita, Miyuki Kunihisa, Takayoshi Yano, Megumi Hamano and Hiromichi Yamazaki

origin of F . × ananassa . We discuss the accuracy of the reported pedigree of cultivars and the relationship between cpDNA haplotype and June yellows, a leaf variegation disorder in strawberry. Materials and Methods We collected fresh leaves from 75

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Mariem Bouhadida, Juan P. Martín, Gennady Eremin, Jorge Pinochet, María Á. Moreno and Yolanda Gogorcena

estimated with a 50-bp ladder marker (Amersham, Piscataway, NJ). Data analysis. The presence or absence of each restriction fragment at each polymorphic site was scored as binary data and used to identify chloroplast haplotypes. Similarity between

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Gal Sapir, Raphael A. Stern, Martin Goldway and Sharoni Shafir

). Research in the three botanical families is brought together in an attempt to explore the S -RNase-mediated GSI system. In GSI, the inhibition of a pollen grain is based on its haploid genotype (termed S -haplotype). SI is manifested if the S