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Title Molecular and Biochemical Studies on Self-Incompatibility of Two Commercially Important Cherry Species (Prunus Avium L. An

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Title Molecular and Biochemical Studies on Self-Incompatibility of Two Commercially Important Cherry Species (Prunus Avium L. An Molecular and biochemical studies on self-incompatibility of Title two commercially important cherry species (Prunus avium L. and P. cerasus L.)( Dissertation_全文 ) Author(s) Yamane, Hisayo Citation 京都大学 Issue Date 2003-01-23 URL https://doi.org/10.14989/doctor.r11106 Right Type Thesis or Dissertation Textversion author Kyoto University 859 Molecular and Biochemical Studies on Self-incompatibility of Two Commercially Important Cherry Species (Prunus avium L. and P. cerasus L.) HISAYO YAMANE Molecular and Biochemical Studies on Self-incompatibility of Two Commercially Important Cherry Species (Prunus avium L. and P. cerasus L.) HISAYO YAMANE Contents General Introduction ------------------------------------------------------------------------------- 1 Chapter 1. Identification and characterization of S-RNases associated with gametophytic self-incompatibility in sweet cherry (Prunus avium L.) 1.1. Introduction -------------------------------------------------------------------- 5 1.2. Materials and Methods ------------------------------------------------------- 5 1.3. Results and Discussion ------------------------------------------------------- 11 1.4. Summary ----------------------------------------------------------------------- 26 Chapter 2. Molecular typing of S-haplotypes in sweet cherry (Prunus avium L.) 2.1. Introduction -------------------------------------------------------------------- 27 2.2. Revisiting the S-haplotype nomenclature of sweet cherry by RFLP analysis 2.2.1. Materials and Methods -------------------------------------------- 28 2.2.2. Results and Discussion -------------------------------------------- 29 2.3. Genomic structure of S-RNases and S-typing by PCR-RFLP analysis 2.3.1. Materials and Methods -------------------------------------------- 34 2.3.2. Results and Discussion -------------------------------------------- 35 2.4. Summary ----------------------------------------------------------------------- 41 Chapter 3. Genetic control of self-incompatibility and self-compatibility in tetraploid sour cherry (Prunus cerasus L.) 3.1. Introduction -------------------------------------------------------------------- 42 3.2. Identification and characterization ofS-RNases in tetraploid sour cherry (Prunus cerasus L.) 3.2.1. Materials and Methods -------------------------------------------- 43 3.2.2. Results and Discussion -------------------------------------------- 47 3.3. The inheritance of S-haplotypes and the segregation of self-incompatibility and self-compatibility phenotypes in 80 F 1 progeny of sour cherry 3.3 .1. Materials and Methods -------------------------------------------- 61 3.3.2. Results and Discussion -------------------------------------------- 63 3.4. Summary ----------------------------------------------------------------------- 68 Chapter 4. Identification of stylar non-S-RNases of sweet cherry: A possible ancestral form of S-RNases in Prunus 4.1. Introduction -------------------------------------------------------------------- 69 4.2. Materials and Methods ------------------------------------------------------- 69 4.3. Results -------------------------------------------------------------------------- 73 4.4. Discussion ---------------------------------------------------------------------- 81 4.5. Summary ------------------------------------------------------------------------ 84 Conclusion and Prospects ------------------------------------------------------------------------- 85 References -------------------------------------------------------------------------------------------- 91 Acknowledgements --------------------------------------------------------------------------------- 104 General Introduction There are two major cultivated cherry species, diploid sweet cherry (Prunus avium L.) and tetraploid sour cherry (P. cerasus L.). Although cherry production fluctuates between years, maximum world production values for sweet and sour cherries are approximately 1,450,000 and 1,600,000 tons, respectively. In Japan, sweet cherry is one of important fruit crop and its production was 21,200 tons covering a production area of about 3,970 ha in 2001 (Source: Ministry of Agriculture, Forestry and Fisheries, Minister's Secretariat, Statistics and Information Department). The production area in Japan is increasing consistently in the last ten years (Source: Zennoh Yamagata). Sweet and sour cherries are reported to have originated in the Near East center, which includes Iran, Iraq, and Syria (Vavilov, 1951). Watkins (1976) suggests that the first diploid Prunus species arose in central Asia, and that sweet and sour cherries were early derivatives of this ancestral Prunus. Ground cherry (P. fruticosa) and sweet cherry are believed to be the progenitor species of sour cherry (Olden and Nybom, 1968). As sweet and sour cherries spread throughout Europe, ecotypes arose within each species that differed in cold hardiness, tree habit, and fruit and leaf characteristics (Kolesnikova, 1975). In Japan, some cherry cultivars were introduced from Europe and USA in the early Meiji era, and have been grown in apple growing regions such as Yamagata. After World War II, sweet cherry improvement started in Japan by breeding program or selection of chance seedlings. In 1990, 17 cultivars were selected for commercial production and now the number of cultivars is increasing with some new cultivars mostly derived from seedlings of the breeding program at Yamagata Prefectural Horticultural Experimental Station. Although the sour cherry market is bigger than the sweet cherry market in the world, sour cherry is rarely grown commercially in Japan because sour cherry suitable to processed use does not meet Japan domestic demand for fresh fruit. Most sour cherries are processed to pie or yogurt, whereas sweet cherries are consumed mostly fresh. Sweet cherry cultivars are predominantly self-incompatible with several self-compatible lines artificially produced through irradiation breeding. Self-incompatibility in sweet cherry was first identified by East and Mangelsdorf 1 (1925) and later determined to be of the gametophytic type with the multiple allelic S-locus (Lewis, 1948). In contrast to sweet cherry, sour cherry cultivars are predominantly self-compatible with several self-incompatible cultivars. Self-incompatibility in sour cherry is characterized by inliibition of pollen tube growth in style, which is typical phenomenon of gametophytic self-incompatibility (Lansari and Iezzoni, 1989). Cherry production is hampered by self-incompatibility because cherries are unable to bear fruit parthenocarpically and fertilization is indispensable for their fruit production. To ensure fruit set, pollinizer trees are inter-planted in the commercial orchard and beehives are introduced. In Japan, alternatively, hand-pollination is practiced although it is a very labor-intensive operation. Self-incompatibility is also a hindrance to breeders wishing to carry out selfing or crossing between cultivars in the same pollination incompatibility groups. One of the breeding objectives of cherries, therefore, is the production of self-compatible cultivars. So far, several self-compatible sweet cherry cultivars, such as 'Stella' and its offspring, have been produced (Lapins, 1970, 1975). Although sour cherry is predominantly self-compatible, self-incompatible selections can result from crosses between two self-compatible parents. Since any successful new sour cherry cultivars would have to be self-compatible, self-incompatibility is also a hindrance to sour cherry breeder. In gametophytic self-incompatibility, the pistil distinguishes between self and non-self pollen based on whether the S-haplotype of the haploid pollen matches either of the two S-haplotypes of the diploid pistil (Figs. 1, 2). Although the gametophytic self-incompatibility had been believed to be controlled by a single genetic locus (S-locus), recent studies revealed that S-locus consists of at least two different genes for the pistil and pollen components that are responsible for self-incompatible reaction (for a review, see McCubbin and Kao, 2000). They are tightly linked to each other as if they were "a single gene". Thus the term haplotype is used to denote variants of the locus, and the term allele is used to denote variants of a given polymorphic gene at the S-locus. When haplotypes match, pollen is recognized by the pistil as self and rejected, whereas if haplotypes differ, pollen is accepted for fertilization. Thus, crosses between two plants are compatible as long as their S-haplotypes differ in one of the two S-haplotypes (Figs. 1,2). 2 The molecular mechanism of gametophytic self-incompatibility has been studied extensively in solanaceous plant species. cDNAs encoding the pistil proteins that were associated with S-haplotype were first cloned from Nicotiana alata (Anderson et aI., 1986). Because the deduced amino acid sequences contained an active site of the fungal RNases T2 (Kawata et aI., 1988) and Rh (Horiuchi et aI., 1988), it was suggested that stylar RNases were involved in the gametophytic self-incompatible reaction (McClure et aI., 1989). Since this finding, the S-gene products in pistils of Solanaceae are called S-RNases. The results from transgenic analyses clearly demonstrated that S-RNase is a pistil determinant for the pollen rejection response in Solanaceae (McCubbin et aI., 1997; Lee et aI., 1994; Murfett et aI., 1994, 1995). Recently, it has been shown that S-RNases are also associated with the gametophytic self-incompatibility of Japanese pear
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