본 연구에서는 분자마커를 이용한 목표형질분리의 효용성을 검토하였다. Ivory seed(IVS) 유전자는 bHLH 단백질을 암호화하고 있고, 꽃의 안토시아닌과 종자의 프로안토시아닌의 발현에 관여하는 애기장대의 TT8 및 페튜니아의 AN1유전자와 상동성을 가지는 유 전자로 나팔꽃의 꽃과 종자의 착색을 촉진하는 것으로 알려져 있다. 아이보리 종자로부터 화색이 full purple(FP), pale purple(PP), magenta(M) 및 pale red(PR)인 4가지 계통의 식물체가 분리되었다. 분자분 석을 통해 이 식물체 모두 ivs 유전자의 변이형질을 지니는 것으로 확인되었다. 분자마커 선발을 위해 고안 된 프라이머를 사용하여 IVS 유전자의 exon 2와 intron 5 영역을 증폭하여, ivs 대립형질인 ivs-stb 및 ivs-pwt을 야생형 IVS로부터 구분할 수 있었다. PCR 결과로 예측한 IVS 유전자의 대립형질 구성 상태는 Southern blot 분석 결과와 정확하게 일치하였다. PP 와 PR은 ivs-stb에 의한 열성 변이체였으며, M과 FP 계통은 ivs-stb 대립형질에 더하여 각각 야생형 IVS 및 ivs-pwt와의 이형접합체로 확인되었다. 본 연구에서 선발된 분자마커는 IVS 유전자의 대립형질 구분에 매 우 유용하였다.
The ω5-gliadins are the major allergens in wheat-dependent excise-induced anaphylaxis (WDEIA). In this study, SDS-PAGE analysis was used to assign the ω5-gliadins (Gli-B1) alleles in thirty two Korean wheat cultivars, compared with eleven standard wheat cultivars for Gli-B1a~m alleles. These results were reconfirmed with their complementary Glu-B3 low-molecular-weight glutenin subunits alleles tightly linked with Gli-B1 locus revealed with 2-DGE in our previous study. As a result, one Gli-B1b, four Gli-B1d, two Gli-B1f, six Gli-B1m and nineteen Gli-B1h varieties were identified. This is the first report on revealing the Gli-B1 alleles in Korean wheat cultivars and represents valuable basic data on wheat allergy, relationship between gliadin and wheat quality, and development of hypo-allergenic wheat.
The spikelet number per panicle (SPP) in rice is a quantitative trait controlled by numerous genes. To identify the alleles of these genes, we conducted sequencing of coding region or UTR of 13 genes which are known for controlling SPP trait, subjected to 48 rice cultivars covering 27 Indica, 17Japonica, and 4 Tongil types. The genes were EP3(ERECT PANICLE 3), RCN1(REDUCED CULM NUMBER 1), RCN2(REDUCED CULM NUMBER 2), FZP(FRIZZY PANICLE), LAX1(LAX PANICLE 1), MOC1(MONOCULM 1) APO1(ABERRANT PANICLE ORGANIZATION 1), APO2(ABERRANT PANICLE ORGANIZATION 2), DEP1(DENSE AND ERECT PANICLE 1), FON1(FLORAL ORGAN NUMBER 1), GHD8(GRAIN NUMBER, PLANT HEIGHT, AND HEADING DATE 8), GN1A(GRAIN NUMBER 1A), and HD1(HEADING DATE1). As a result, 1 synonymous SNP was found in EP3, RCN1 and RCN2. In LAX1 and MOC1, 1 non-synonymous SNP was identified. But none of SNP was found from the coding region of FZP. Moreover, 3, 6, 7, 8, 9 and 10 alleles were detected in APO1 and APO2, FON1, GHD8, GN1A, DEP1, and HD1 respectively. The multiple regression analysis revealed that GHD8, DEP1 and HD1 had strong effects on spikelet number of primary branch. In addition, HD1 had strong effects on spikelet number of secondary branch, and for total spikelet number per panicle as well. This work was supported by a grant from the Next-Generation BioGreen 21 Program (Plant Molecular Breeding Center No. PJ008125), Rural Development Administration, Republic of Korea.
Allelic variations in glutenin and puroindolines of 26 Korean wheat cultivars were evaluated to determine their effects on the physicochemical properties of flour and quality of white salted noodles. Cultivars carrying Pina-D1b and Pinb-D1b exhibited a coarser particle size of wheat flour and a higher ash and damaged starch content than those with Pina-D1a and Pinb-D1a. Glu-B1b, Glu-D1f, Glu-B3d and Pina-D1a alleles exhibited lower protein content than other alleles. Glu-A1c, Glu-B1b, Glu-D1f Glu-B3d, Glu-B3i and Pinb-D1b alleles appeared to be related to a lower SDS-sedimentation volume than other alleles. In dough rheological properties, Glu-A1a and Glu-D1d alleles showed a longer mixing time on the mixograph and maximum dough height but Glu-A3e and Glu-B3i alleles had a lower mixing time on the mixograph and a lower maximum dough height than other alleles at Glu-1 and Glu-3 loci. Regarding the quality of white salt noodles, about 10% of the variations in the hardness of cooked noodles were explained by Glu-A1 and Glu-B3 loci. Hardness rankings of cooked noodles were Glu-A1a > Glu-A1c > Glu-A1c at the Glu-A1 locus. Glu-B3h showed higher cooked noodle hardness (5.10 N) than other alleles at the Glu-B3 locus (< 4.66 N).
Protein related parameters of pan bread and white salted noodles prepared from 26 Korean wheat cultivars and 6 commercial and imported wheat flours were evaluated to elucidate the relationship between rheological properties and end-use characteristics and to determine the effects of Glu-1 and Glu-3 alleles on those characteristics in Korean wheat cultivars. SDS-sedimentation volume based on protein weight was positively correlated with mixograph parameters and maximum height of dough and also positively correlated with bread loaf volume, crumb firmness and springiness of cooked noodles. Within Glu-1 loci, 1 or 2* subunit and 5 + 10 subunits showed longer mixingtime, higher maximum height of dough, and larger loaf volume than null allele, 2.2 + 12, and 2 + 12 subunits. Cultivars with 13 + 16 subunits at Glu-B1 locus showed higher protein content and optimum water absorption of mixograph than cultivars with 7 + 8 subunits. At Glu-3 loci, Glu-A3d showed longer mixing time than Glu-A3e, and Glu-B3d and Glu-B3h had stronger mixing properties than Glu-B3i. Glu-B3h had higher loaf volume and hardness of cooked noodles than Glu-B3d. Glu-D3a had lower protein content than Glu-D3c, and Glu-D3b showed stronger mixing properties than Glu-D3a. Glu-D3c showed lower hardness of cooked noodles than others.
Allelic variations in glutenin and puroindolines of Korean wheat cultivars evaluated to determine the effects of allelic variations on physico-chemical properties of flour and qualities of white salted noodles. In grain hardness and flour yield, Pinb-D1b had higher hardness index and flour yield than Pinb-D1a alleles. Glu-B1b and Glu-D1f also had lower hardness index than other alleles at the same locus and Glu-A1c, Glu-A3e and Glu-B3i alleles showed lower flour yield than other alleles. In flour compositions, Pina-D1b and Pinb-D1b showed higher particle size, ash and damaged starch content and lower lightness of wheat flour than Pina-D1a and Pinb-D1a. Glu-A1c, Glu-B1b, Glu-D1f, Glu-B3d and Glu-B3i showed lower particle size of flour than other alleles at Glu-1 and Glu-B3 locus. Korean wheats with Glu-B1f, Glu-D1a and Glu-B3b alleles had higher damaged starch content and lower lightness of flour than wheats other alleles at the same locus. Glu-B1b, Glu-D1f, Glu-B3d and Pina-D1a showed lower protein content and Glu-A1c, Glu-B1b, Glu-D1f Glu-B3d, Glu-B3i and Pinb-D1b showed lower SDS-sedimentation volume than other alleles. Hardness of cooked noodles ranked as Glu-A1a > Glu-A1c > Glu-A1c at Glu-A1 locus. Glu-B3h showed higher hardness of cooked noodles (5.10 N) than other alleles at Glu-B3 locus (< 4.66 N).
Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for F_1-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called "functional" markers for allelic selection of the restorer-of-fertility gene (Rfo) in F_1-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating F_1-hybrid cultivar development in radish.
To investigate the genetic variation of high-and low-molecular-weight glutenin subunits (BMW-GS and LMW-GS), granule-bound starch synthase I (GBSSI) and puroindoline in 24 Korean wheat cultivars. At the BMW-GS compositions, three Glu-A1 alleles, five Glu-B1 alleles and three Glu-D1 alleles were identified. The high frequency of alleles at each locus was Glu-A1c allele (15 cultivars), Glu-B1b allele (16 cultivars) and Glu-D1f allele (16 cultivars). Four alleles were identified at the Glu-A3 and Glu-B3 loci and three at Glu-D3 locus and Glu-A3d, Glu-B3d and Glu-D3a were mainly found at each Glu-3 locus. Glu-A3d, Glu-B3d, Glu-D3b or c (4 cultivars, respectively) and Glu-A3d, Glu-B3d, Glu-D3a and Glu-A3c, Glu-B3d or h, Glu-D3a (3 cultivar, respectively) were predominantly found in Korean wheats. At the GBSS compositions, 2 waxy wheat cultivars, Shinmichal and Shinmichal1, showed null alleles on the Wx loci and other cultivars were wild type in GBSS compositions. At the puroindoline gene compositions, Korean wheat cultivars carried 3 genotypes, which 10 cultivars (41.7%) were Pina-D1a and Pinb-D1a, 11 cultivars (45.8%) had Pina-D1a and Pinb-D1b and 3 cultivars (12.5%) carried Pina-D1b and Pinb-D1a. These genetic variations could present the information to improve flour and end-use quality in Korean wheat breeding programs.