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  • Author or Editor: Ke Wang x
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Peach (Prunus persica) is an important fruit crop worldwide with several thousand cultivars. Cultivar discrimination and hybrid authentication are often required in peach breeding and can be achieved by applying various molecular markers including simple sequence repeat (SSR). In this study a total of 2146 expressed sequence tag (EST)–SSR loci were detected with the 10,737 EST sequences retrieved from the NCBI. A total of 49 EST-SSR markers, including 24 simple ones with a motif comprising of tri-, tetra-, penta-, hexanucleotides, and 25 compound ones, were selected and then primers were designed. Following conventional polymerase chain reaction (PCR) specificity control and sequence authentication, as well as fluorescence-based PCR product size and stutter band evaluation, 37 EST-SSR markers with correct amplification and without stutter band interference were validated. Among them, 14 were polymorphic in 18 closely related peach accessions, with polymorphism information content (PIC) ranging from 0.0994 to 0.3750. The 18 peach accessions can be distinguished using nine polymorphic markers, with the exception of ‘Shangshandayulu’ and ‘Xipu 1’, both being bud sports from ‘Yulu’. The clustering of the accessions as well as the fingerprint profiles supported the authentication of the hybrids. These EST-SSR markers are useful for peach breeding research.

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Crown gall disease caused by Agrobacterium tumefaciens affects a wide range of horticultural plants, and has no effective treatment. During the evaluation of crown gall resistance of peach germplasm resources, we observed enhanced resistance to subsequent invasion that was activated by virulence of A. tumefaciens in two peach cultivars. To further verify the phenotype observed in field experiments, systemic acquired resistance (SAR)-related salicylic acid (SA) and PR1 genes were investigated. The levels of SA were elevated in two cultivars, and these high levels were maintained for 35 days postinoculation. Compared with mock-inoculated controls, eight of the 22 candidate PpPR1 genes in A. tumefaciens-inoculated samples were significantly upregulated and three were downregulated in response to inoculation with A. tumefaciens. These data suggested that SA-induced SAR was activated in two peach cultivars by virulent A. tumefaciens infection. In addition, the eight induced PpPR1 genes can be used as molecular markers in defense studies in peach.

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Anthocyanins are important molecules that are responsible for fruit color formation and are also beneficial to human health. To date, numerous structural and regulatory genes associated with anthocyanin biosynthesis in peach (Prunus persica) have been reported based on linkage analysis. In this study, we sought to identify further genes associated with anthocyanin content in peach by conducting a genome-wide association analysis of 129 peach accessions to detect markers associated with the trait. Significant association signals were detected when anthocyanin content was considered a qualitative character but not when it was considered a quantitative trait. We detected an association region located between 11.7 and 13.1 Mb in chromosome 1, a region in which only 133 of 146 genes have previously been functionally annotated. Gene ontology annotation of the genes in this region showed that membrane-associated genes (including one gene encoding a chloride channel protein and 17 sugar transport/carrier-associated genes) were significantly enriched, and we focused on these in subsequent analyses. Based on in vitro induction of anthocyanins in fruit flesh using different exogenously applied sugars and subsequent culture, we found that the expression level of 3 of the 18 membrane-associated genes, Prupe.1G156300, Prupe.1G156900, and Prupe.1G157000, increased during induction treatment. Furthermore, during the fruit development period of a white-fleshed and a red-fleshed peach cultivar, the expression of one gene encoding a transmembrane sugar transport protein was observed to be positively correlated with anthocyanin biosynthesis. These results will facilitate understanding of the molecular mechanism of anthocyanin biosynthesis in peach.

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Excised leaf sections of lance coreopsis cultured on Murashige Skoog (MS) medium produced adventitious shoots in response to BA. When the combinations of 0, 0.5, 1, or 2 μm NAA with 0, 5, 10, 20, or 40 μm BA were tested, shoots were induced by any of the four BA concentrations used in the medium, regardless of the presence of NAA. The average number of shoots formed per leaf section ranged from 1.4 to 4.3 seven weeks after culture initiation. Roots were induced at the base of individual shoots on the same regeneration medium when cultures were kept longer than 7 weeks. The rooted plants were transferred successfully into soil. The regenerated plants had the same growth and flowering characteristics as the seed-grown plants. Chemical names used: benzyladenine (BA); naphthaleneacetic acid (NAA).

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Hydrangea macrophylla is the most popular species in the genus Hydrangea because of its large and brightly colored inflorescences. Since the early 1900s, numerous cultivars with showy flowers have been selected. Although many H. macrophylla cultivars have been developed, cold hardiness is still the major limitation to their outdoor use. Hydrangea arborescens is a small attractive shrub or subshrub native to northeastern parts of the United States, which is valued for its hardiness. Interspecific breeding of H. arborescens and H. macrophylla has been tried, but putative hybrid seedlings either died at an early stage or were not verified. We made successful hybridizations between H. macrophylla ‘Blue Diamond’ and H. arborescens ‘Annabelle’ and used in vitro ovary culture to produce viable plants. Hybrids were intermediate in appearance between parents, but variable in leaves, inflorescences, and flower color. The success of this hybridization was confirmed by six simple sequence repeat (SSR) genetic markers. The maternal chromosome number was 36, and the paternal number was 38. Chromosome counts of hybrids indicated that nearly half of them were aneuploids. Male fertility of progeny was evaluated by fluorescein diacetate staining of pollen. Twelve out of 14 hybrids (85.7%) had male fertility. We documented the first flowering progeny of H. macrophylla and H. arborescens, suggesting an effective beginning to a cold hardiness breeding program.

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The root-knot nematode (RKN) Meloidogyne incognita can cause severe crop loss in economically important Prunus species like peach (P. persica), almond (P. communis), plum (P. salicina), and apricot (P. armeniaca). Some peach rootstock, including Nemaguard (P. persica), Nemared (P. persica), and Myrobalan plum (P. cerasifera), display significant resistance to RKN. We present a genetic linkage map constructed by using simple sequence repeat (SSR) and sequence-related amplified polymorphism (SRAP) for a peach backcross population (190 individuals) of RKN-resistant ‘Honggengansutao’ (P. kansuensis) and susceptible ‘Bailey’ (P. persica). Degenerate primers designed from conserved motifs of known plant resistance gene (R) products were used to amplify genomic DNA sequences. Twenty-two resistance gene analog (RGA) sequences were selected from 48 RGAs with open-reading frames to design sequence-tagged site markers. The linkage map of ‘Honggengansutao’ is composed of 138 loci (30 SSRs, 102 SRAPs, five RGAs, and one morphological marker for RKN resistance) assigned to eight linkage groups. The map covers 616 cM of the peach genome with an average marker spacing of 4.9 cM. The five RGAs were mapped to Groups 2, 7, and 8. One gene (designated PkMi) involved in resistance to RKN was mapped to Group 2 (which also includes the known RKN-resistance RMia gene). BLASTN analysis mapped all RGAs to the peach genome sequence. The map constructed in the study will aid future rootstock breeding with marker-assisted selection to identify additional candidate RGA sequences.

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Many reports indicate that an abundance of really interesting new gene (RING) play key roles in regulating defense responses against abiotic and biotic stresses in plants. In this study, the cloning and functional characterization of a RING gene, MaRING2, in banana (Musa acuminata) fruit are reported. MaRING2 belongs to the NEP1-interacting protein (NIP) RING-H2 finger protein family. Gene expression profiles revealed that MaRING2 was cold responsive and induced by abscisic acid (ABA) treatment during cold storage. In this study, the MaRING2 under control of the Cauliflower mosaic virus 35S (CaMV 35S) promoter was transformed to tobacco (Nicotiana benthamiana) using agrobacterium (Agrobacterium tumefaciens)-mediated transformation. The resultant MaRING2-overexpressing transgenic plants (35S:MaRING2) exhibited significantly increased tolerance to low temperatures and were hypersensitive to exogenous ABA in terms of germination and early seedling growth. In addition, overexpression of MaRING2 enhanced the expression of stress-responsive genes under normal (before cold stress) or cold conditions. These results demonstrate the biological role of MaRING2 in conferring cold tolerance. Taken together, these results suggest that MaRING2, a C3H2C3-type RING protein, is a positive regulator of the ABA-dependent stress response.

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Amplified fragment length polymorphism (AFLP) analyses were used to assess genetic diversity among 30 genotypes of watermelon [Citrullus lanatus (Thunb.) Mansf.] representing a broad genetic base, including breeding lines and commercial germplasm. Eight AFLP primer combinations selected from 64 primer combinations were polymophic. The polymorphism was 13.0% to 31.9% within the 28 cultivars examined, and 45.3% to 64.2% among all the genotypes. Each genotype could be successfully distinguished based on AFLP scoring. Cluster grouping of accessions based on the AFLP analysis was consistent with that from classification by pedigrees and ecotypes.

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The phenotypic expression and inheritance of the rolC gene in the transgenic plants of Salpiglossis sinuata L. were investigated. The chasmogamous salpiglossis plants with solid yellow flower color (ccrrDD) were transformed with Agrobacterium tumefaciens strains LBA4404 and EHA101 carrying rolC, GUS, and NPTII genes via a leaf disc co-cultivation system. The transgenic plants were shorter in plant height, produced more branches with a compact growth habit, and developed smaller flowers and narrower leaves as compared to the control plant. While the transgenic plants showed the same corolla color and color shades as the parental line, they became male sterile. A backcross between a male-sterile transgenic plant (ccrrDD plus rolC) and a nontransformed red-flowering line (ccRRDD) produced a progeny with red flower color and the same altered growth habit as the transgenic female parent. Only 4 out of 32 plants in this progeny population showed the negative GUS staining as well as the non transgenic phenotype. These results suggest that at least two copies of the rolC gene were integrated into one homologous chromosome pair during transformation and that a cross-over event may have occurred during meiosis.

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Petal anthocyanins were systematically identified and characterized by high-performance liquid chromatography (HPLC)–electrospray ionization–mass spectrometry (MS) coupled with diode array detection among nine wild herbaceous peony (Paeonia L.) species (15 accessions). Individual anthocyanins were identified according to the HPLC retention time, elution order, MS fragmentation patterns, and by comparison with authentic standards and published data. Six main anthocyanins, including peonidin-3,5-di-O-glucoside, peonidin-3-O-glucoside-5-O-arabinoside (Pn3G5Ara), peonidin-3-O-glucoside, pelargonidin-3,5-di-O-glucoside, cyanidin-3,5-di-O-glucoside, and cyanidin-3-O-glucoside (Cy3G), were detected. In addition to the well-known major anthocyanins, some minor anthocyanins were identified in herbaceous peony species for the first time. Detection of the unique anthocyanins cyanidin-3-O-glucoside-5-O-galactoside and pelargonidin-3-O-glucoside-5-O-galactoside in both Paeonia anomala L. and P. anomala ssp. veitchii (Lynch) D.Y. Hong & K.Y. Pan indicated these two species should belong to the same taxon. Pn3G5Ara was found only in European wild species and subspecies suggesting different metabolic pathways between European and Chinese accessions. Anthocyanins conjugated with galactose and arabinose were observed in the genus Paeonia for the first time. The North American species, Paeonia tenuifolia L., had high Cy3G content in flower petals. This anthocyanin composition is distinct from the anthocyanin composition in Asian and European species and possibly is responsible for the vivid red coloration in flowers.

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