Suk-Man Kim

Map-based Cloning and Characterization of the BPH18 Gene from Wild Rice Conferring Resistance to Brown Planthopper (BPH) Insect Pest

Brown planthopper (BPH) is a phloem sap-sucking insect pest of rice which causes severe yield loss. We cloned the BPH18 gene from the BPH-resistant introgression line derived from the wild rice species Oryza australiensis. Map-based cloning and complementation test revealed that the BPH18 encodes CC-NBS-NBS-LRR protein. BPH18 has two NBS domains, unlike the typical NBS-LRR proteins. The BPH18 promoter::GUS transgenic plants exhibited strong GUS expression in the vascular bundles of the leaf sheath, especially in phloem cells where the BPH attacks. The BPH18 proteins were widely localized to the endo-membranes in a cell, including the endoplasmic reticulum, Golgi apparatus, trans-Golgi network, and prevacuolar compartments, suggesting that BPH18 may recognize the BPH invasion at endo-membranes in phloem cells. Whole genome sequencing of the near-isogenic lines (NILs), NIL-BPH18 and NIL-BPH26, revealed that BPH18 located at the same locus of BPH26. However, these two genes have remarkable sequence differences and the independent NILs showed differential BPH resistance with different expression patterns of plant defense-related genes, indicating that BPH18 and BPH26 are functionally different alleles. These findings would facilitate elucidation of the molecular mechanism of BPH resistance and the identified novel alleles to fast track breeding BPH resistant rice cultivars.

Identification for quantitative trait loci controlling grain shattering in rice

Seed shattering is an important factor causing loss of grain yield before and during rice harvest. In the present study, the quantitative trait loci regarding shattering scale, breaking tensile strength (BTS) and abscission layer (AL), the parameters evaluating seed shattering habit by hand gripping, a digital force gauge and observation on AL, respectively, were identified by using an doubled haploid line (DHL) population from a cross between a loose-shattering type Tongil variety, ‘Samgang’, and a moderately difficult shattering japonica variety, ‘Nagdong’. Eight QTLs consisted in four QTLs for shattering scale, two QTLs for AL, each one QTL for pulling and bending strength were detected on six chromosomes, respectively. Among them, Qss1 with flanking markers RM6696 and RM476 explained 31% of phenotype variation in shattering scale. Furthermore, two new QTLs controlling shattering habit, Qss5-2 and Qal5-1, were located on chromosome 5 at the interval 5028–5037 and 5021-RM289. They explained 10% and 12% of phenotype variations, respectively. A total of eleven digenic epistatic loci were identified for four parameters. The identification of QTLs affecting seed shattering habits is favorable to thoroughly dissect the genetic mechanism of the shattering habit and to apply for marker-assisted selection in rice breeding system of specific regions.

Genetic analysis and QTL mapping for grain chalkiness characteristics of brown rice (Oryza sativa L.)

Grain chalkiness is one of the important appearance qualities in rice marketing. But it is a complex trait, controlled by polygenes and easily influenced by the environment. Genetic analysis and QTL detection was carried out on six characteristics of grain chalkiness consisting of the percentage of chalkiness (PGC), white belly (PWB) and white core grains (PWC), and the area of chalkiness (CA), white belly (WBA) and white core (WCA) in brown rice. A total of 16 main-effect QTLs associated with chalkiness characteristics of brown rice were mapped on seven chromosomes over two years. Among them,qPGC7.1 andqPWB7.2 were simultaneously located on chromosome 7 flanked by 7038 and 7042 at LOD scores 4.34 and 3.76, whileqPWC2.1 andqWCA2.1 were simultaneously located on chromosome 2 flanked by RM492 and RM324 with LOD scores of 2.50 and 3.39. Twelve epistatic combinations were detected for five chalkiness characteristics except for CA. Results indicated that WBA was mainly influenced by the additive effects of main-effect QTLs. PGC and PWC were affected by the effects of epistatic QTLs and the interactions between additive-by-additive effects and the environment. The effects of epistatic QTLs and the main-effect QTLs played important roles on CA, PWB and WCA. For the genetic improvement of grain chalkiness in breeding system, more attention should be paid to epistatic effects and the additive effects of main-effect QTLs.

QTL detection and MAS selection efficiency for lipid content in brown rice (Oryza sativa L.)

Rice lipid content as one of important ingredients of functional food and industrial products has become an entirely new target in the rice breeding programs worldwide. A genetic linkage map spanning 12 rice chromosomes with an average interval of 10.51 cM between markers was created using 172 DNA markers, which intended to elucidate genetic basis of lipid content in brown rice by QTL detection. Eight QTLs related to lipid content with LOD from 2.52 to 7.86 were mapped on chromosome1, 2, 3, 5, 6, 7 and 9 using a doubled haploid (DH) population from a cross of ‘Samgang/Nagdong’ with field experiments for five years. Two QTLs of qLC5.1 and qLC6.1 in the intervals 5014-5024 and 6011-RM19696 were repeatedly detected over four years at average LOD scores of 4.85 and 4.21, respectively. Five of eight QTLs tend to increase the lipid content from ‘Samgang’ alleles. Epistatic and environmental effects played important roles and explained 42.20% of phenotype variations. Three QTLs of qLC6.1, qLC7.1 and qLC9.1 collectively explained much than 27% of phenotype variations and increased 0.25% of lipid content and, showed much than 85% of selection efficiency for the lines with high lipid contents in the F7 population from a cross of ‘Samgang/Nagdong’. Thus it provides the sufficient possibility to realize QTLs pyramiding and to promote process of rice breeding.

Comparison and analysis of main effects, epistatic effects, and QTL × environment interactions of QTLs for agronomic traits using DH and RILs populations in rice.

Two genetic linkage maps based on doubled haploid (DH) and recombinant inbred lines (RILs) populations, derived from the same indica-japonica cross ‘Samgang × Nagdong’, were constructed to analyze the quantitative trait loci (QTLs) affecting agronomic traits in rice. The segregations of agronomic traits in RILs population showed larger variations than those in DH population. A total of 10 and 12 QTLs were identified on six chromosomes using DH population and seven chromosomes using RILs population, respectively. Three stable QTLs including pl9.1, ph1.1, and gwp11.1 were detected through different years. The percentages of phenotypic variation explained by individual QTLs ranged from 8 to 18% in the DH population and 9 to 33% in the RILs population. Twenty-three epistatic QTLs were identified in the DH population, while 21 epistatic QTLs were detected in the RILs population. Epistatic interactions played an important role in controlling the agronomic traits genetically. Four significant main-effect QTLs were involved in the digenic interactions. Significant interactions between QTLs and environments (QE) were identified in two populations. The QTLs affecting grain weight per panicle (GWP) were more sensitive to the environmental changes. The comparison and QTLs analysis between two populations across different years should help rice breeders to comprehend the genetic mechanisms of quantitative traits and improve breeding programs in marker-assisted selection (MAS).

Identification and characterization of QTLs and QTL interactions for Macro- and Micro-elements in rice (Oryza sativa L.) grain

Improvement of the macro- and micro-elements density of rice (Oryza sativa L.) is gradually becoming a new breeding objective. In this study, the genomic regions associated with potassium, calcium, magnesium and iron content in rice grain were identified and characterized by using a doubled haploid (DH) population. Fifty-six simple sequence repeat (SSR) and one hundred and twelve sequence tagged site (STS) markers were selected to construct the genetic linkage map of the DH population with a full length of 1808.3cM scanning 12 rice chromosomes. Quantitative trait loci (QTLs) were detected, and QTL effects and QTL interactions were calculated for five traits related to macro- and micro-elements in the DH population from a cross between `Samgang` (Tongil) and `Nagdong` (Japonica). Twelve QTLs were located on five chromosomes, consisting of two QTLs for potassium, three QTLs for calcium, two QTLs for magnesium, one QTL for iron content and four QTLs for the ratio of magnesium to potassium (Mg/K). Among them, qca1.1 was detected on chromosome 1 with an LOD value of 8.58 for calcium content. It explained 27% of phenotype variations with increasing effects from `Samgang` allele. Furthermore, fifteen epistatic combinations with significant interactions were observed on ten chromosomes for five traits, which totally accounted for 4.19% to 12.72% of phenotype variations. The screening of relatively accurate QTLs will contribute to increase the efficiency of marker-assisted selection (MAS), and to accelerate the establishment of near-isogenic lines (NILs) and QTL pyramiding.