DNA Marker Based Diversity Across Rice Genotypes and Advanced Breeding Lines Bred for Temperate India

Background: Characterization and evaluation of plant genetic resources play an important role for their utilization in the crop improvement programmes. Methods and results: This study entails the agro-morphological, cooking quality and molecular characterization of 51 genotypes / advance breeding lines of rice from Kashmir Himalayas. Significant variability was observed for all agro-morphological and cooking quality traits among all the studied genotypes. Cluster analysis using UPGMA method divided the genotypes into two major clusters having 15 and 36 genotypes. Thirty eight genotypes screened using 24 SSR markers detected 48 alleles with 2.0 alleles per locus and an average polymorphism information content (PIC) of 0.37. High polymorphism information content (PIC) values was observed for the primers RM263 (0.67), RM159 (0.59) and RM333 (0.50). Furthermore, out of 38 SSR markers screened on 192 temperate rice germpalsm lines, R4M17 accurately differentiated indica and temperate japonica genotypes amplifying 220 bp and 169bp, respectively. Accordingly, 15 genotypes were reported as indica and 28 temperate japonica in addition to 149 genotypes as intermediate types. Conclusion: The information on marker-based diversity and performance based on cooking quality and agronomic traits helped to select the most divergent lines for crossing and also the analysis was useful to generate information on indica - japonica classification of our germplasm.

Breeding Temperate Japonica Rice Varieties Adaptable to Tropical Regions: Progress and Prospects

Temperate japonica rice is mainly cultivated in temperate regions. Many temperate japonica varieties have a superior grain quality that is preferred in Northeast Asian countries such as Japan, Korea, and China. The changes in consumers’ preferences in Southeast Asia and Western countries has contributed to increasing the demand for temperate japonica. Most temperate japonica varieties developed in temperate regions typically exhibit extra-early flowering under the short-day conditions in the tropics, which usually results in severely reduced yields. Since 1992, we have been developing temperate japonica varieties that can adapt to tropical environments to meet the increasing demand for temperate japonica rice, having released six varieties in the Philippines. Especially, the yield of one of the temperate japonica varieties, Japonica 7, was comparable to the yields of leading indica varieties in the Philippines. Here, we discuss the current breeding initiatives and future plans for the development of tropical-region-bred temperate japonica rice.

Genomic Variation in Korean japonica Rice Varieties

Next-generation sequencing technologies have enabled the discovery of numerous sequence variations among closely related crop varieties. We analyzed genome resequencing data from 24 Korean temperate japonica rice varieties and discovered 954,233 sequence variations, including 791,121 single nucleotide polymorphisms (SNPs) and 163,112 insertions/deletions (InDels). On average, there was one variant per 391 base-pairs (bp), a variant density of 2.6 per 1 kbp. Of the InDels, 10,860 were longer than 20 bp, which enabled conversion to markers resolvable on an agarose gel. The effect of each variant on gene function was predicted using the SnpEff program. The variants were categorized into four groups according to their impact: high, moderate, low, and modifier. These groups contained 3524 (0.4%), 27,656 (2.9%), 24,875 (2.6%), and 898,178 (94.1%) variants, respectively. To test the accuracy of these data, eight InDels from a pre-harvest sprouting resistance QTL (qPHS11) target region, four highly polymorphic InDels, and four functional sequence variations in known agronomically important genes were selected and successfully developed into markers. These results will be useful to develop markers for marker-assisted selection, to select candidate genes in map-based cloning, and to produce efficient high-throughput genome-wide genotyping systems for Korean temperate japonica rice varieties.

Rice (Oryza sativa L.) breeding for novel black short grain associated with yield, cooking quality, high nutrition and antioxidant potential derived from indica black rice x japonica white rice

The combination of the trend of Japanese food consumption with the health benefits of black rice is in high demand for rice consumers in Thailand. For this challenge, incorporation of desirable traits from temperate japonica white rice, Akitakomachi and tropical indica black rice, Riceberry was performed by pedigree selection with maker assisted selection (MAS) and these desirable traits have been presented in breeding progenies. The three candidate lines showed highly favorable agronomic characteristics and a high grain yield, with short grains and good cooking quality, similar to japonica rice, in a tropical climate. In addition, these lines showed black coloration of the pericarp, indicating high nutritional value (including Fe, Zn, Vit B6, Vit E and folate levels) and phytochemical, antioxidant and antidiabetic activities, similar to those of the Riceberry parent. In terms of the sensory testing of unpolished rice, two breeding lines (69-1-1 and 72-4-3) showed higher scores than their parents. However, only 69-1-1 was identified as japonica type according to its genetic background. Therefore, this breeding program, involving the crossing of a temperate japonica white rice with a tropical indica black rice, can create novel black short grain rice variety adapted to a tropical environment, similar to japonica-type rice.

Landraces of temperate japonica rice have superior alleles for improving culm strength associated with lodging resistance

Lodging can reduce grain yield and quality in cereal crops including rice (Oryza sativa L.). To achieve both high biomass production and lodging resistance, the breeding of new cultivars with strong culms is a promising strategy. However, little is known about the diversity of culm strength in temperate japonica rice and underlying genetic factors. Here, we report a wide variation of culm strength among 135 temperate japonica cultivars, and some landraces having the strongest culms among these cultivars. The genome-wide association study (GWAS) identified 55 quantitative trait loci for culm strength and morphological traits, and revealed several candidate genes. The superior allele of candidate gene for culm thickness, OsRLCK191, was found in many landraces but had not inherited to the modern improved cultivars. Our results suggest that landraces of temperate japonica rice have unutilized superior alleles for contributing future improvements of culm strength and lodging resistance.

Development of 454 New Kompetitive Allele-Specific PCR (KASP) Markers for Temperate japonica Rice Varieties

Temperate japonica rice varieties exhibit wide variation in the phenotypes of several important agronomic traits, including disease resistance, pre-harvest sprouting resistance, plant architecture, and grain quality, indicating the presence of genes contributing to favorable agronomic traits. However, gene mapping and molecular breeding has been hampered as a result of the low genetic diversity among cultivars and scarcity of polymorphic DNA markers. Single nucleotide polymorphism (SNP)-based kompetitive allele-specific PCR (KASP) markers allow high-throughput genotyping for marker-assisted selection and quantitative trait loci (QTL) mapping within closely related populations. Previously, we identified 740,566 SNPs and developed 771 KASP markers for Korean temperate japonica rice varieties. However, additional markers were needed to provide sufficient genome coverage to support breeding programs. In this study, the 740,566 SNPs were categorized according to their predicted impacts on gene function. The high-impact, moderate-impact, modifier, and low-impact groups contained 703 (0.1%), 20,179 (2.7%), 699,866 (94.5%), and 19,818 (2.7%) SNPs, respectively. A subset of 357 SNPs from the high-impact group was selected for initial KASP marker development, resulting in 283 polymorphic KASP markers. After incorporation of the 283 markers with the 771 existing markers in a physical map, additional markers were developed to fill genomic regions with large gaps between markers, and 171 polymorphic KASP markers were successfully developed from 284 SNPs. Overall, a set of 1225 KASP markers was produced. The markers were evenly distributed across the rice genome, with average marker density of 3.3 KASP markers per Mbp. The 1225 KASP markers will facilitate QTL/gene mapping and marker-assisted selection in temperate japonica rice breeding programs.