Cai-lin Wang

QTL Mapping for Grain Size Traits Based on Extra-Large Grain Rice Line TD70

Abstract Grain size traits, including grain length, grain width and grain thickness, are controlled by quantitative trait loci (QTLs). Many QTLs relating to rice grain size traits had been reported, but their control mechanisms have not yet been elucidated. A recombinant inbred line (RIL) population of 240 lines, deriving from a cross between TD70, an extra-large grain size japonica line with 80 g of 1000-grain weight, and Kasalath, a small grain size indica variety, were constructed and used to map grain size QTLs to a linkage map by using 141 SSR markers in 2010 and 2011. Five QTLs for grain length, six for grain width and seven for grain thickness were detected distributing over chromosomes 2, 3, 5, 7, 9 and 12. Seven QTLs, namely qGL3.1 , qGW2 , qGW2.2, qGW5.1 , qGW5.2 , qGT2.3 and qGT3.1 , were detected in either of the two years and explained for 56.19%, 4.42%, 29.41%, 10.37%, 7.61%, 21.19% and 17.06% of the observed phenotypic variances on average, respectively. The marker interval RM1347–RM5699 on chromosome 2 was found common for grain length, grain width and grain thickness; qGL3.1 and qGT3.1 were mapped to the same interval RM6080–RM6832 on chromosome 3. All 18 QTL alleles were derived from the large grain parent TD70. Most of the QTLs mapped in the present study were found the same as the genes previously cloned ( GW2 , GS3 or qGL3 , GW5 and GS5 ), and several were the same as the QTLs ( GS7 and qGL-7 ) previously mapped. Three QTLs, qGL2.2 on chromosome 2, qGW9 and qGT9 on chromosome 9, were first detected. These results laid a foundation for further fine mapping or cloning of these QTLs.

Genetic Analysis of Rice False Smut Resistance Using Mixed Major Genes and Polygenes Inheritance Model

Abstract To study the inheritance of the resistance to rice false smut [Ustilaginoidea virens (Cooke) Takahashi], a population of 157 recombinant inbred lines (RILs) of F10 derived from the cross between resistant cultivar IR28 (Oryza sativa supsp. indica) and susceptible landrace Daguandao (O. sativa supsp. japonica) was analyzed using the mixed major genes and polygenes inheritance model. The indicator of resistance was disease rate index scored based on percentage of infected grains, which was investigated 3 weeks after artificial inoculation. The results showed that the resistance to rice false smut was controlled by the mixed 2 major genes and polygenes model (model E-1–3). The 2 major genes had equivalent additive effect of 11.41, and their heritability was about 76.67%. The heritability of polygenes was about 22.86%. Both effects of major genes and polygenes should be considered in the resistance breeding to rice false smut.

Cluster Analysis on Japonica Rice （Oryza sativa L.） with Good Eating Quality Based on SSR Markers and Phenotypic Traits

Abstract Diversity of 60 conventional japonica rice accessions with good eating quality at home and abroad was analyzed using SSR molecular markers, agronomic traits and taste characteristics. A total of 290 alleles were detected in the 60 accessions at 72 SSR loci with the high similarity coefficients varying between 0.600 and 0.924. The loci on chromosome 5 showed the greatest value in average allele number. Additionally, most of the SSR loci could detect 3 to 4 alleles. An UPGMA dendrogram based on the cluster analysis of the genetic similarity coefficients showed that the grouping trend of part of the rice accessions was geographic-related and most of the rice accessions in Jiangsu Province, China were clustered together. Furthermore, many domestic accessions from south and north origins in China were close to the foreign japonica rice varieties, as proved by their pedigree origin from the foreign high-quality sources. For taste characteristics, part of the accessions with excellent taste were clearly clustered into one category though they came from different geographical regions, which indicates that taste characteristics of some varieties were mainly genetically determined. In addition, the agronomic traits of japonica rice with good taste might be closely related with their geographical origins, but the relationship between superior taste characteristics and agronomic traits should be further clarified.

Transferring Translucent Endosperm Mutant Gene Wx-mq and Rice Stripe Disease Resistance Gene Stv-bi by Marker-Assisted Selection in Rice (Oryza sativa)

A high-yielding japonica rice variety, Wuyunjing 7, bred in Jiangsu Province, China as a female parent was crossed with a Japanese rice variety Kantou 194, which carries a rice stripe disease resistance gene Stv-bi and a translucent endosperm mutant gene Wx-mq. From F2 generations, a sequence characterized amplified region (SCAR) marker tightly linked with Stv-bi and a cleaved amplified polymorphic sequence (CAPS) marker for Wx-mq were used for marker-assisted selection. Finally, a new japonica rice line, Ning 9108, with excellent agronomic traits was obtained by multi-generational selection on stripe disease resistance and endosperm appearance. The utilization of the markers from genes related to rice quality and disease resistance was helpful not only for establishing a marker-assisted selection system of high-quality and disease resistance for rice but also for providing important intermediate materials and rapid selection method for good quality, disease resistance and high yield in rice breeding.

Inheritance Analysis and QTL Mapping of Rice Starch Viscosity (Rapid Visco Analyzer Profile) Characteristics

The rice starch viscosity characteristics, which can be indicated by Rapid Visco Analyzer profile (RVA profile), have been proved useful for the evaluation of cooking and eating quality in rice breeding program. To study the inheritance of the RVA profile, an F2 population of Wuyujing 3/Aichi 106 was used. The results indicated that the peak viscosity (PKV) was a typical quantitative character, and the hot paste viscosity (HPV), cool paste viscosity (CPV), setback viscosity (SBV), breakdown viscosity (BDV), and consistence viscosity (CSV) might be controlled by one major gene and several minor genes. To elucidate the genetic basis of the paste viscosity characteristics, a recombinant inbred line (RIL) population derived from Nikken 2/Milyang 23 and its genetic linkage map were used to map the QTLs controlling RVA profiles in 2005 and 2006. A total of 34 QTLs distributed on chromosomes 1, 2, 3, 4, 6, 7 and 8 were detected, including 19 and 15 QTLs in 2005 and 2006, respectively. Eight QTLs were both detected in the two years, qHPV6, qCPV6, qCSV6, qSBV6, and qBDV6 were located on chromosome 6, while qHPV2, qCSV2, and qCPV2 were on chromosome 2.

Comparative transcriptome analysis reveals molecular response to salinity stress of salt-tolerant and sensitive genotypes of indica rice at seedling stage

Abiotic stresses, such as salinity, greatly threaten the growth and productivity of plants. Rice (Oryza sativa L.) is one of the most important food crops, as well as a monocot model for genomic research. To obtain a global view of the molecular response to salinity stress, we conducted a leaf transcriptome analysis on rice seedlings. Two cultivars of rice subspecies indica, including the salt-tolerant genotype Xian156 and the salt-sensitive genotype IR28, were used in the present study. Eighteen RNA libraries were obtained from these two genotypes at three timepoints (0 h, 48 h and 72 h) after applying salinity stress. We obtained the reference-guided assembly of the rice transcriptome, which resulted in 1,375 novel genes, including 1,371 annotated genes. A comparative analysis between genotypes and time points showed 5,273 differentially expressed genes (DEGs), of which 286 DEGs were only found in the tolerant genotype. The Disease resistance response protein 206 and TIFY 10 A were differentially expressed, which were validated by quantitative real-time PCR. The differentially expressed genes identified through the mRNA transcriptome, along with the structure, provide a revealing insight into rice molecular response to salinity stress and underlie the salinity tolerance mechanism between genotypes.

Development of Simple Functional Markers for Low Glutelin Content Gene 1 (Lgc1) in Rice (Oryza sativa)

Abstract Rice with low glutelin content is suitable as functional food for patients affected by kidney failure. Low glutelin-content gene Lgc1 in rice has a 3.5-kb deletion between two highly similar glutelin genes GluB4 and GluB5, which locates on the short arm of chromosome 2. To improve the selection efficiency in low glutelin-content rice breeding, two molecular markers designated as InDel-Lgc1-1 and InDel-Lgc1-2 were developed to detect the low glutelin-content gene Lgc1. A double PCR detection indicated that combined use of the two markers could easily distinguish the genotypes of Lgc1 from different rice varieties. Therefore, as a simple and low-cost technique, the molecular marker could be widely used to identify different varieties with Lgc1 gene and applied in marker-assisted selection of low glutelin-content rice.

Development of New InDel Marker to Detect Genotypes of Rf-1a Conferring Fertility Restoration of BT-Type Cytoplasmic Male Sterility in Rice

Restorer line breeding is an important approach to enhance the heterosis and improve the yields of japonica hybrid rice. To improve the selection efficiency of restorer lines for BT-type cytoplasmic male sterility (CMS) in japonica rice, a functional marker InDel-Rf-1a based on the difference of nucleotide sequence in Rf-1a locus between BT-type CMS lines and restorer lines was developed to detect the genotypes of different rice materials. Conventional indica rice varieties, restorer and maintainer lines without 574 bp deletion could restore the fertility for BT-type CMS in japonica rice. By contrast, most conventional japonica rice varieties except Aichi 106 and Yijing 12, with genotype of rf-1arf-1a showed the 574 bp deletion maintained sterility for BT-type CMS lines. To further verify the effect of genotyping detection in Rf-1a locus, this marker was also used to amplify the genomic DNA in different japonica rice restorer lines, CMS lines, hybrids and F2 segregation population, and three genotypes in Rf-1a locus could be distinguished distinctly. Therefore, the marker InDel-Rf-1a could be widely used for genetic identification and marker-assisted selection (MAS) in breeding japonica restorer lines.

Detection of QTL for Cold Tolerance at Bud Bursting Stage Using Chromosome Segment Substitution Lines in Rice (Oryza sativa)

The cold tolerance at the bud bursting stage (CTB) was evaluated at 5°C by using a set of 95 chromosome segment substitution lines (CSSLs) derived from an indica rice 9311 and a japonica rice Nipponbare with a genetic background of 9311. The result showed that six CSSLs had slightly stronger effect on CTB than 9311. Total four quantitative trait loci (QTLs) for CTB were preliminary mapped on chromosomes 5 and 7 by substitution mapping. qCTB-5-1, qCTB-5-2 and qCTB-5-3 were mapped in the region of RM267–RM1237, RM2422–RM6054 and RM3321–RM1054, which were 21.3 cM, 27.4 cM and 12.7 cM in genetic distance on rice chromosome 5, respectively. qCTB-7 was mapped in a 6.8-cM region of RM11–RM2752 on rice chromosome 7.

A novel semidwarf mutant 02428ha in rice is caused by reduced gibberellin sensitivity

The rice (Oryza sativa L.) elongated uppermost internode (eui) gene effectively results in a recessive tall plant type. In eui background, we found a mutant 02428ha that exhibited a semidwarf phenotype. Because semidwarf varieties possess high yield potential, this mutant may be useful for hybrid rice breeding. The present study aimed to characterize the dwarfing mechanism in the mutant. Microscopic observation revealed that the cell length in the uppermost internode was reduced compared to the wild type. The mutant showed a decreased sensitivity to exogenous gibberellin A3 (GA3) and gibberellin (GA) biosynthesis inhibitor, uniconazole. Meanwhile, it contained a high level of endogenous gibberellin A1 (GA1). These results indicated that the semidwarf phenotype of 02428ha was caused by impairment of GA signal transduction rather than by block in GA biosynthesis. Furthermore, expression analysis of genes involved in the GA signaling pathway showed that transcript of GIBBERELLIN INSENSITIVE DWARF2 (GID2) was down-regulated in the mutant. Sequence analysis also revealed a 15-base pair (bp) insertion between base 594 and 595 in the coding region of GID2. Consequently, the reason for down-regulation expression of GID2 or sequence variation of GID2 might play roles in the phenotype of 02428ha.