Comparison of phenotypic versus marker-assisted background selection for the SUB1 QTL during backcrossing in rice

Marker assisted backcrossing has been used effectively to transfer the submergence tolerance gene SUB1 into popular rice varieties, but the approach can be costly. The selection strategy comprising foreground marker and phenotypic selection was investigated as an alternative. The non-significant correlation coefficients between ranking of phenotypic selection and ranking of background marker selection in BC₂F₁, BC₃F₁ and BC₃F₂ generations indicated inefficiency of phenotypic selection compared to marker-assisted background selection with respect to recovery of the recipient genome. In addition, the introgression size of the chromosome fragment containing SUB1 was approximately 17 Mb, showing the effects of linkage drag. The significant correlation coefficient between rankings of phenotypic selection with the percentage of recipient alleles in the BC₁F₁ generation suggested that background selection could be avoided in this generation to minimize the genotyping cost. The phenotypically selected best plant of the BC₃F₁ generation was selfed and backcross recombinant lines were selected in the resulting BC₃F₄ generation. The selection strategy could be appropriate for the introgression of SUB1 QTL in countries that lack access to high-throughput genotyping facilities.     

Confirming a major QTL and finding additional loci responsible for field resistance to brown spot (Bipolaris oryzae) in rice

Brown spot is a devastating rice disease. Quantitative resistance has been observed in local varieties (e.g., ‘Tadukan’), but no economically useful resistant variety has been bred. Using quantitative trait locus (QTL) analysis of recombinant inbred lines (RILs) from ‘Tadukan’ (resistant) × ‘Hinohikari’ (susceptible), we previously found three QTLs (qBS2, qBS9, and qBS11) that conferred resistance in seedlings in a greenhouse. To confirm their effect, the parents and later generations of RILs were transplanted into paddy fields where brown spot severely occurred. Three new resistance QTLs (qBSfR1, qBSfR4, and qBSfR11) were detected on chromosomes 1, 4, and 11, respectively. The ‘Tadukan’ alleles at qBSfR1 and qBSfR11 and the ‘Hinohikari’ allele at qBSfR4 increased resistance. The major QTL qBSfR11 coincided with qBS11 from the previous study, whereas qBSfR1 and qBSfR4 were new but neither qBS2 nor qBS9 were detected. To verify the qBSfR1 and qBSfR11 ‘Tadukan’ resistance alleles, near-isogenic lines (NILs) with one or both QTLs in a susceptible background (‘Koshihikari’) were evaluated under field conditions. NILs with qBSfR11 acquired significant field resistance; those with qBSfR1 did not. This confirms the effectiveness of qBSfR11. Genetic markers flanking qBSfR11 will be powerful tools for marker-assisted selection to improve brown spot resistance.     

Cold tolerance at the early growth stage in wild and cultivated rice

The present study was conducted to understand the pattern of variation and the genetic bases for cold tolerance at the early growth stage in Asian rice. The genetic variation was investigated at the germination, plumule and seedling stages among 57 strains including cultivated rice (Oryza sativa ssp. indica and ssp. japonica) and its wild progenitor (Oryza rufipogon). The significant differentiation of cold tolerance was observed among the taxonomically divided groups. At the germination stage, both indica and japonica subspecies tended to be more tolerant than O. rufipogon, whereas at the plumule and seedling stages, ssp. japonica tended to be more tolerant than ssp. indica and O. rufipogon. Furthermore, in cold tolerance at the plumule stage, the clinal variation across the latitude of origins was observed within O. rufipogon and ssp. japonica, suggesting that the current pattern of variation seems to have been shaped by both their phylogenetic histories and on-going adaptation to the local environments. QTL analysis between O. sativa ssp. japonica (tolerant) and O. rufipogon (susceptible) revealed five putative QTLs for cold tolerance at the plumule and seedling stages but not at the germination stage. Substitution mapping was also carried out to precisely locate the two major QTLs for cold tolerance at the plumule stage, which could be used for improvement of tolerance to cold stress in ssp. indica.     

Variation in cooking and eating quality traits in Japanese rice germplasm accessions

The eating quality of cooked rice is important and determines its market price and consumer acceptance. To comprehensively describe the variation of eating quality in 183 rice germplasm accessions, we evaluated 33 eating-quality traits including amylose and protein contents, pasting properties of rice flour, and texture of cooked rice grains. All eating-quality traits varied widely in the germplasm accessions. Principal-components analysis (PCA) revealed that allelic differences in the Wx gene explained the largest proportion of phenotypic variation of the eating-quality traits. In 146 accessions of non-glutinous temperate japonica rice, PCA revealed that protein content and surface texture of the cooked rice grains significantly explained phenotypic variations of the eating-quality traits. An allelic difference based on simple sequence repeats, which was located near a quantitative trait locus (QTL) on the short arm of chromosome 3, was associated with differences in the eating quality of non-glutinous temperate japonica rice. These results suggest that eating quality is controlled by genetic factors, including the Wx gene and the QTL on chromosome 3, in Japanese rice accessions. These genetic factors have been consciously selected for eating quality during rice breeding programs in Japan.     

Cytoplasmically controlled cold tolerance in a cytoplasmic-genetic male sterile line of rice

Summary The cytoplasmic-genetic male sterile line, Krishna-A with Kalinga-I cytoplasm was developed in rice through repeated backcrossing the completely pollen sterile F₁ of the cross Kalinga-I/Krishna with the recurrent male parent cv. Krishna. The germination percentage and root-shoot length in Krishna-A at 12°C and 9°C indicated a higher degree of tolerance to low temperature than the cold tolerant female parent Kalinga-I. The high yielding cold tolerant variety Kalinga-I was developed from a cross involving the cold tolerant variety Dunghansali and the high yielding variety IR 8. It is inferred that tolerance to low temperature during germination in Krishna-A was inherited from the cytoplasm of cv. Dunghansali through Kalinga-I besides male sterility. The cytoplasmic control of cold tolerance is reported here for the first time in rice literature. The male sterile line Krishna-A because of its tolerance to low temperature would be suitable for the development of hybrid rice especially for areas where low temperature is a problem during germination.     

Variation in heading date conceals quantitative trait loci for other traits of importance in breeding selection of rice

To identify quantitative trait loci (QTLs) associated with the primary target traits for selection in practical rice breeding programs, backcross inbred lines (BILs) derived from crosses between temperate japonica rice cultivars Nipponbare and Koshihikari were evaluated for 50 agronomic traits at six experimental fields located throughout Japan. Thirty-three of the 50 traits were significantly correlated with heading date. Using a linkage map including 647 single-nucleotide polymorphisms (SNPs), a total of 122 QTLs for 38 traits were mapped on all rice chromosomes except chromosomes 5 and 9. Fifty-eight of the 122 QTLs were detected near the heading date QTLs Hd16 and Hd17 and the remaining 64 QTLs were found in other chromosome regions. QTL analysis of 51 BILs having homozygous for the Koshihikari chromosome segments around Hd16 and Hd17 allowed us to detect 40 QTLs associated with 27 traits; 23 of these QTLs had not been detected in the original analysis. Among the 97 QTLs for the 30 traits measured in multiple environments, the genotype-by-environment interaction was significant for 44 QTLs and not significant for 53 QTLs. These results led us to propose a new selection strategy to improve agronomic performance in temperate japonica rice cultivars.     

Identification of qRL7, a major quantitative trait locus associated with rice root length in hydroponic conditions

Root system development is an important target for improving yield in rice. Active roots that can take up nutrients more efficiently are essential for improving grain yield. In this study, we performed quantitative trait locus (QTL) analyses using 215 recombinant inbred lines derived from a cross between Xieqingzao B (XB), a maintainer line with short roots and R9308, a restorer line with long roots. Only a QTLs associated with root length were mapped on chromosomes 7. The QTL, named qRL7, was located between markers RM3859 and RM214 on chromosome 7 and explained 18.14–18.36% of the total phenotypic variance evaluated across two years. Fine mapping of qRL7 using eight BC₃F₃ recombinant lines mapped the QTL to between markers InDel11 and InDel17, which delimit a 657.35 kb interval in the reference cultivar Nipponbare. To determine the genotype classes for the target QTL in these BC₃F₃ recombinants, the root lengths of their BC₃F₄ progeny were investigated, and the result showed that qRL7 plays a crucial role in root length. The results of this study will increase our understanding of the genetic factors controlling root architecture, which will help rice breeders to breed varieties with deep, strong and vigorous root systems.     

Combination of stem elongation ability with submergence tolerance in rice

Summary A study was conducted with six F₂ populations to test the possibility of combining submergence tolerance and stem elongation ability into a single genotype of rice (Oryza sativa L.). Submergence tolerance and stem elongation ability could be combined in the same genotype if strongly submergence tolerant genes are present in submergence tolerant parents.     

Inheritance of salt tolerance in rice

Summary The genetic behavior of salt tolerance was studied in artificially salinized conditions at the International Rice Research Institute.Divergent selection, carried out at a salinity level where the ECe was 15.2 mmhos/cm at 25 C in F₃ lines from two crosses confirmed the effects of salt tolerance on F₄ progeny with realized heritability values of 0.39 and 0.62, respectively.In a cross between two tolerant cultivars there was clear over-dominance for tolerance, despite the high environmental fluctuation which resulted in a low genetic response as indicated by a low but significant repeatability of 0.20–0.25, and many progeny lines more tolerant than the parents were recovered. The superior tolerance of these progenies compared to the parents was confirmed subsequently at 3 different salt levels. In the same experiment a cross between tolerant and susceptible cultivars produced some progeny of comparable tolerance with tolerant sources.In a 6×6 diallel cross experiment with two tolerant, moderate, and susceptible varieties each, both general and specific combining ability were significant.The findings indicate the possibility of breeding rices more tolerant than existing tolerant cultivars through cumulative crosses of tolerant cultivars. Further improvement can be attained by crossing highly tolerant lines with donors of good agronomic traits and pest and disease resistance.     

QTL analysis of seed dormancy in rice (Oryza sativa L.)

Effective cumulative temperature (ECT) after heading would be a more reasonable parameter for seed sampling of pre-harvest sprouting/seed dormancy (SD) tests in segregating populations than the days after flowering. SD is an important agronomic trait associated with grain yielding, eating quality and seed quality. To identify genomic regions affecting SD at different grain-filling temperatures, and to further examine the association between SD and ECT during grain-filling, 127 double haploid (DH) lines derived from a cross between ZYQ8 (indica)/JX17 (japonica) by anther culture were analyzed. The quantitative trait loci (QTLs) and their digenic epistasis for SD were identified using a molecular linkage map of this population. A total of four putative QTLs for SD (qSD-3, qSD-5, 6 and 11) were detected on chromosomes 3, 5, 6 and 11, together explaining 41.4% of the phenotypic variation. Nine pairs of digenic epistatic loci were associated with SD on all but chromosome 9, and their contributions to phenotypic variation varied from 2.87%–8.73%. The SD QTL on chromosome 3 was identical to the QTLs found in other mapping populations with different genetic backgrounds, which could be a desirable candidate for gene cloning and marker-assisted selection in rice breeding.     

QTL analysis for panicle characteristics in temperate japonica rice

To understand the genetic background of panicle characteristics in temperate japonica rice (Oryza sativa L.), we genetically analyzed DH lines derived from a cross between two temperate japonica rice cultivars, ‘Akihikari’ and ‘Koshihikari’,in 1996 and 1997. Four traits of panicle characteristics, number of primary branches per panicle (NPB), number of secondary branches per panicle (NSB), average number of spikelets on one primary branch (NSP)and average number of spikelets on one secondary branch (NSS), in 212 DH lines were measured, and the interval mapping of QTLs for these traits was carried out using169 DNA markers with an LOD threshold of2.5. Five, three and one putative QTLs for NPB, NSB and NSS were identified,respectively, and no QTLs relating to NSP appeared. The percentages in total phenotypic variation explained by all putative QTLs for NPB were 35.5%: and43.8% in 1996 and 1997, respectively. All putative QTLs for NSB accounted for 35.5%and 27.5% of total phenotypic variation in1996 and 1997, respectively. The QTLs identified in this study will be useful intemperate japonica rice breeding for improved spikelet yield. This revised version was published online in August 2006 with corrections to the Cover Date.     

Parameters of cold tolerance in common vetch

Summary The cold tolerance of 16 common vetch (Vicia sativa L.) strains was assessed by means of survival rate after direct freezing. Additionally, several seed and seedling parameters, germination in solution with osmotic pressure, and electrolyte exosmosis from shoots and from roots of seedlings were determined. The U.S. cv's Nova II, Vanguard and Warrior and the Turkish cv. Sari Elçi exhibited survival rates higher than 75% after direct freezing test, compared to less than 40% in the remaining entries. Simple correlation analysis showed that only electrolyte exosmosis from shoots after freezing was negatively correlated with survival rate.     

Characterization and fine mapping of thermo-sensitive chlorophyll deficit mutant1 in rice (Oryza sativa L.)

Chlorophyll content is one of the most important traits controlling crop biomass and economic yield in rice. Here, we isolated a spontaneous rice mutant named thermo-sensitive chlorophyll deficit 1 (tscd1) derived from a backcross recombinant inbred line population. tscd1 plants grown normally from the seedling to tiller stages showed yellow leaves with reduced chlorophyll content, but showed no significant differences after the booting stage. At temperatures below 22°C, the tscd1 mutant showed the most obvious yellowish phenotype. With increasing temperature, the yellowish leaves gradually turned green and approached a normal wild type color. Wild type and tscd1 mutant plants had obviously different chloroplast structures and photosynthetic pigment precursor contents, which resulted in underdevelopment of chloroplasts and a yellowish phenotype in tscd1. Genetic analysis indicated that the mutant character was controlled by a recessive nuclear gene. Through map-based cloning, we located the tscd1 gene in a 34.95 kb region on the long arm of chromosome 2, containing two BAC clones and eight predicted candidate genes. Further characterization of the tscd1 gene is underway. Because it has a chlorophyll deficit phenotype before the tiller stage and little influence on growth vigor, it may play a role in ensuring the purity of hybrids.     

A novel procedure for raising rice seedlings and its potential application for mass-screening of rice strains for salt tolerance

Summary A soilless culture system is described whereby all the necessary steps of raising rice seedlings, including seeding, surface-sterilization, germination and growing of seedlings, can be conveniently achieved within a small plastic container. The system is especially suited for experiments with massive number of entries and can e.g. be used for screening of rice genotypes for salt tolerance.