Rice eating quality is considered to be one of the top priorities in determining the agronomical value of rice. Thus the rapid evaluation of eating quality at early breeding generations in breeding programs for better eating quality is of great importance. However, it has been limited due to the complex nature of eating quality and the absence of standard evaluation method. In our previous study, we developed a evaluation method with a set of DNA markers that allows to predict the eating quality for japonica rices. Here we successfully developed another marker set for the eating quality of indica rices. We used multiple regression analysis to test 54 markers, which were preselected for their possible association with eating quality, using 24 indica varieties with different palatability scores. Of these markers, eighteen markers were found to be significantly associated with palatability according to sensory evaluation. Accordingly, a marker set in the model regression equation with a high R2 (0.997) was formulated to estimate indica rice palatability. Validation suggests that markers and the statistical parameters formulated by the equation could be a potential tool to predict the palatability of cooked Indonesian indica rice and could be reliable in developing country-dependent model equations for eating quality. 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.
Rice hulls remain closed throughout the ripening period to maintain internal humidity of the grains. An Open-hull sterile mutant was induced by N-methyl-N-nitrosourea(MNU) treatment on Sinsunchalbyeo rice, a japonica type. This mutant showed open hulls even in the ripening stages and fully mature grains. In addition, several altered characteristics were observed, including of narrowed palea, decreased grain size, partial pollen sterility and erect panicle. Microscopic analysis showed that the palea was positioned slightly inside the lemma, and the size of palea decreased in the mutant. Genetic analysis of F2 and F3 segregation populations derived from the cross between the Open-hull sterile mutant (Oryza sativa ssp. japonica) and Milyang23 (O. sativa ssp. indica) indicated that the Open-hull trait was controlled by a single recessive allele. The fine-mapping with STS (sequence tagged site) markers revealed that the mutant gene was located on the short arm of chromosome 3. We were able to narrow it down until 30.6Kb where three candidate genes were found.
As the market demand on functionality rice has been increasing, embryo rice in which embryo residue remains even after milling has come to comsumers’ attention because rice embryo contains several functionality components. Consequently, development of rice varieties for higher rate of embryo adhesion to grains after milling has become one of the breeding objectives for quality improvement. In this study, we observed embryo dent of 49 commercial varieties and analyzed the relationship between embryo dent and grain size and shape. Embryo dent of rice grains varied 0.27 (Keunnun)~0.59 (Daerip 1) mm. Varieties Jinbu, Jinbo, Heugseol, Obong, Unkwang, and Cheongnam showed relatively deeper embryo dent, suggesting that they will be applicable in breeding for embryo rice. Embryo dent was correlated positively with grain width (r=0.53**) and grain size(r=0.34*), and negatively with grain width/length ratio (r= -0.38**). Strategies for breeding embryo rice were discussed in relation to embryo dent, grain size and shape.
Plant height is an important agronomic trait that affects grain yield. Previously, we reported a novel semi-dominant dwarf mutant, D-h, derived from chemical mutagenesis using N-methyl-N-nitrosourea(MNU) on a japonica rice cultivar, Hwacheongbyeo. In this study, we cloned the gene responsible for the dwarf mutant using the map-based approach. Fine mapping revealed that the mutant gene was located on the short arm of chromosome 1 in a 48 kb region. Sequencing of the candidate genes and rapid amplification of cDNA ends-polymerase chain reaction(RACE-PCR) analyses identified the gene, d-h, which encodes a protein of unknown function, but whose sequence is conserved in other cereal crops. Real-time (RT)-PCR analysis and promoter activity assay showed that the d-h gene was primarily expressed in the nodes and the panicle. In the D-h mutant plant, the gene was found to carry a 63-bp deletion in the ORF region, which was confirmed to be directly responsible for the mutant’s gain of a functional phenotype by subsequent transgenic experiments. Since the mutant plants exhibit a defect in the GA response, but not in the GA synthetic pathway, it appears that the d-h gene may be involved in a GA signaling pathway.
The architecture of rice panicle is primarily determined by the arrangement of branches and spikelets, and it directly affects grain yield. We identified a mutant for panicle apical abortion from a japonica cultivar Hwacheongbyeo treated with N-methyl-N-nitrosourea. Under normal growth conditions, the mutant had multiple abnormal phenotypes, such as a slight reduction in plant height, narrow and dark green leaf blades, and small erect panicles with clear panicle apical abortion compared to the wild-type plants. Genetic analysis revealed that the panicle apical abortion was controlled by a single recessive gene, which is tentatively designated as paa. The paa gene was fine mapped at an interval of 71 kb flanked by STS markers aptn3 and S6685-1 at the long arm of chromosome 4. Sequence analysis of the candidate genes within the delimited region showed a single base-pair change corresponding to an amino acid substitution from glycine to glutamic acid. We expect that the paa gene will be a clue to uncover the molecular mechanism of panicle apical abortion and to maintain the panicle identity for grain yield in rice breeding programs.
근래 쌀의 건강기능성에 대한 관심이 고조되면서 백미상태에서도 배아의 일부가 부착된 배아미의 이용을 위한 기술적 품종적 연구가 착수되고 있다. 본 연구는 다양한 종자형태를 보이는 49개의 품종에 대하여 현미 상태에서의 배아함몰도 및 종자형태와의 관련성에 대해 조사 분석한 것이다. 배아함몰도는 0.27(큰눈벼)~0.59(대립벼1) mm의 변이를 보였다. 49개의 품종 중 진부, 진보, 흑설, 오봉, 운광, 청남 등이 배아함몰도가 상대적으로 높은 품종이었다. 배아함몰도는 현미의 폭과는 정의 상관관계가 있었고 장폭비와는 부의 상관관계를 보여주었다. 배아함몰도가 높은 품종 육성을 위한 방향을 논의하였다.
Two sugary mutants, Hwacheong sugary-1 (Hsu1) and Hwacheong sugary-2 (Hsu2) were obtained by chemical mutagenesis from japonica cultivar, Hwacheongbyeo. Sugary mutants exhibited wrinkled and translucent grain with high soluble sugar content. In addition, amber-colored endosperm of sugary mutants was loosely packed due to abnormal starch granules compared to densely packed wild-type. Especially, the grain of Hsu2 mutant was less wrinkled than that of Hsu1, thus Hsu2 can be polished easily. Previous studies reveal that su1 mutant was resulted from mutation in gene for a debranching enzyme, isoamylase but the sequence of the mutated gene has not been identified. To identify the sequence of sugary genes, the map-based cloning strategy was applied. The genetic study revealed that the phenotype of Hsu2 mutant was controlled by two recessive genes. Interestingly, one of the genes was located on chromosome 8 at the position of isoamylase which was known as su-1. This indicates that mutation in isoamylase gene causes sugary endosperm characteristics. However we found different mutation points between the Hsu1 and Hsu2. The point mutation in Hsu1 was occurred at 10th exon whereas the other mutation related with Hsu2 was occurred at 15th exon. As mentioned above, the Hsu2 mutant has less wrinkled shape and less soluble sugar content than the Hsu1 mutant. Thus, we hypothesize that the other gene controlling Hsu2 mutant phenotype may have a role in weakening the effect of the su-1. Further study on the other gene associated with the Hsu2 phenotype is in progress.
Grain size is one of the most important trait determining yield in cereal crops, apart from number of grains per panicle, number of panicles per plant and 1000 seed weight. Other than grain characteristics, plant architecture is another very important factor influencing yield by affecting the amount plant surface area directly exposed to the sun light. Erect panicle is important morphological characteristic which helps in enhancing the yield by allowing sun light to fall directly on leaves unlike curved panicle which blocks sunlight and consequently reduce photosynthesis. A small round grain and erect panicle mutant was obtained by treating Hwacheong rice (japonica) with MNU (N-methyl-N-nitrosourea) chemical mutagen. Through bulked segregant analysis (BSA) using STS (Sequence-Tagged Sites) and SS-STS (Sub-species Specific Sequence-Tagged Site) markers we located the mutated gene on the long arm of chromosome 7 and narrowed down candidate region to 168.75kbp through fine mapping. Mutant manifested characteristics like reduced grain size and plant height, dense and erect panicle and relatively erect plant compared to the wild type. When we crossed the mutant with its parent (Hwacheong), F1 panicle and grain characteristics showed intermediate phenotype, therefore, we concluded that wild type allele of this gene shows incomplete dominance. Scanning electron microscopy(SEM) result shows that increase in width of mutant grain, which changes its shape, is due to increase in width of glume cells. Phenotypic examination shows that dense and erect panicle phenotype is result of reduction in length of rachis, primary and secondary branch.
The rice sucrose synthase 3 (RSUS3) localized predominantly inrice seed endosperm may play an important role in the starch filling in the milky stage of rice seed. As the genetic diversity at this locus is not known yet, forty three rice varieties/accessions were objected to amplify full sequence of the RSUS3 to examine the distribution of DNA polymorphisms. A total of 254 sequence variants, including 82, 150 and 22SNP sand indels, were successfully identified in whole length of 7,733bp sequence comprising promoter, exon and intron, and 3’ down stream non transcribed region(NTR). Eleven haplotypes were distinguishable among 43 rice varieties based on the nucleotide variation on the three defined regions (5’NTR, transcript and 3’NTR). The promoter region showed the occurrence of a base change on a cis-element which might involve a functional role of the motif in seed-specific expression. Non random process seemed to be acted in the genetic diversity of RSUS3geneamongricegermplasmusedinthisstudy. The analysis of polymorphism sites indicated a history of eleven minimum recombination mostly occurred in the transcribed region. This result might provide an insight for a clasditic approach for establishing future genetic association studies of RSUS3locus.
Sucrose synthase 3 which is a third active gene present in rice, is localized predominantly in rice endosperm. This sucrose synthase 3 may play an important role in the starch filling in the milky stage rice seed, probably involving in the starch physicochemical properties. As the genetic diversity at this locus is little informed, forty three rice consisting of japonica, indica and Oryza rufipogon were targeted to amplify full sequence of sucrose synthase 3 to examine the frequency and distribution of nucleotide polymorphism. Total of 755 all sequence variants detected, 491 single nucleotide polymorphisms (SNPs) and 264 indels were successfully identified in 7618 bp of sequence containing the sucrose synthase 3 transcript, promoter and 3' non-transcribed region. The frequency of nucleotide changes and indels were high, on average one polymorphism per 15.5 bp and one indel per 28.9 bp with 11 sequence-based haplotypes distinguishable among the varieties and lines. Both the frequency of nucleotide changes and indels were frequent in non-coding region, but rare in coding region. Sequencing a polymorphism region in the promoter showed one base change on one of cis-element from T (CATGCATA to A (CATGCACA) that might implicate in seed specificity. The presence of a high number of haplotype shared by a few varieties indicated a little information on linkage disequilibrium.