Hyo Guen Park

Three AFLP markers tightly linked to the genic male sterility ms 3 gene in chili pepper (Capsicum annuum L.) and conversion to a CAPS marker

Genic male sterility (GMS) has long been used as a tool for hybrid seed production in chili pepper (Capsicum annuum L.). We developed DNA markers linked to the GMS ms3 gene in a segregating population using bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) techniques. The segregating population was subjected to BSA-AFLP with 512 primer combinations. Three AFLP markers (Eagg/Mccc276, Eagc/Mctt178, and Ecag/Mtgc204) were identified as tightly linked to the ms3 locus. Among them, we converted the AFLP marker Ecag/Mtgc204 to the cleavage amplified polymorphic sequence (CAPS) marker, named GMS3-CAPS, based on sequencing analysis of internal and flanking regions for the markers between male-fertile and sterile plants. This marker will be useful for pepper breeding using the GMS system.

Genetics of resistance to five strains of turnip mosaic virus in Chinese cabbage

SummaryInheritance of resistance to turnip mosaic virus (TuMV) strains C1, C2, C3, C4, and C5 in Chinese cabbage (Brassica campestris subsp. pekinensis) was evaluated using monoclonal antibodies. Crosses were made between a resistant line, ‘0–2’, and four susceptible line. ‘Seoul’ (SE),‘SSD31’ (SS), ‘Cheongbang’ (CH), and ‘Yaki 1 ho’ (YA), to determine the inheritance of resistance of ‘0–2’ in different genetic backgrounds. Resistance to TuMV was controlled by a single dominant gene or double dominant genes depending on the strain and cross. The resistance genes of ‘0–2’ were modified by susceptible parents such that a single dominant gene was involved in the ‘SS×0–2’ combination, but double dominant genes in the ‘SE×0–2’ against TuMV-C3 or TuMV-C5. ELISA tests using inoculated and noninoculated leaves in the same plant suggested that the dominant resistance genes inhibit virus movement rather than virus multiplication.

A CAPS marker associated with the partial restoration of cytoplasmic male sterility in chili pepper (Capsicum annuum L.)

Cytoplasmic male sterility (CMS), an economically important trait for hybrid seed production in many crops, is a maternally inherited trait in which a plant fails to produce functional anthers, pollen grains, or male gametes. It has long been reported that the restoration of CMS in chili pepper is controlled by a major nuclear gene termed restorer-of-fertility (Rf), along with several modifiers and some environmental factors. In this study, we identified the partial restoration (pr) locus related to the fertility restoration of CMS, demonstrated the inheritance of the trait, and developed a CAPS marker closely linked to the locus. The partially restored plant had normal anthers that produced a mix of normal and aborted pollen grains that stuck tightly to the anther wall, even after dehiscence. This trait was expressed only when the pepper plant had the sterile (S) cytoplasm and homozygous recessive pr alleles. A total of 768 AFLP primer combinations were screened, and bulked segregant analysis (BSA) was performed by preparing two pools of eight Pr/Pr (fully fertile) and eight pr/pr (partially fertile) plants, respectively, selected from the 87 individuals of the F2 segregating population. Of the eight Pr-linked AFLP markers that were identified, E-AGC/M-GCA122 and E-TCT/M-CCG116 were the closest to the locus, estimated at about 1.8 cM in genetic distance. E-AGC/M-GCA122 was converted into a CAPS marker, PR-CAPS, based on the sequences of the internal and flanking regions of the AFLP fragment. This PR-CAPS marker could be useful in selecting fully fertile lines (Pr/Pr) and eliminating partially fertile (pr/pr) and potential (Pr/pr) lines in segregant populations during the development of new inbred restorer lines.

Production of Somatic Hybrids between Satsuma Mandarin (Citrus unshiu) and Navel Orange (Citrus sinensis) by Protoplast Fusion

We have regenerated altotetraploid plants that are interspecific somatic hybrids between Citrus sinensis Osbeck cv. Yoshida navel orange and Citrus unshiu Marc cv. Okitsu satsuma mandarin. Protoplasts isolated from ‘Yoshida’ leaves were chemically fused with call us-derived protoplasts from ‘Okitsu’. After 6 months of culture, 102 plants were obtained. These hybrids were identified by differential leaf morphology, DNA fluorescence intensity, and DNA analysis. Ploidy analysis via the flow cytometry revealed that 15 of the 102 plants were tetraploids, with the rest being diploids that morphologically resembled their mesophyll parent. SRAP analysis confirmed that 9 of the tetraploid plants were allotetraploid somatic hybrids. These will be utilized as a possible pollen parents for improving seedy citrus cultivars, e.g., ponkan, mandarin, lemon and kumquat, in order to produce triploid seedless hybrids.