Zhen Zhu

Development of Chromosome Segment Substitution Lines Derived from Backcross between Two Sequenced Rice Cultivars, Indica Recipient 93-11 and Japonica Donor Nipponbare

Chromosome segment substitution lines (CSSLs) are powerful tools for detecting and precisely mapping quantitative trait loci (QTLs) and evaluating gene action as a single factor. In this study, 103 CSSLs were produced using two sequenced rice cultivars: 93-11, an elite restorer indica cultivar as recipient, and Nipponbare, a japonica cultivar, as donor. Each CSSL carried a single chromosome substituted segment. The total length of the substituted segments in the CSSLs was 2,590.6xa0cM, which was 1.7 times of the rice genome. To evaluate the potential application of these CSSLs for QTL detection, phenotypic variations of seed shattering, grain length and grain width in 10 CSSLs were observed. Two QTLs for seed shattering and three for grain length and grain width were identified and mapped on rice chromosomes. The results demonstrate that CSSLs are excellent genetic materials for dissecting complex traits into a set of monogenic loci. These CSSLs are of great potential value for QTL mapping and plant marker-assisted breeding (MAB).

Physiological character and gene mapping in a new green- revertible albino mutant in rice.

A green-revertible albino mutant-Qiufeng M was found from the japonica rice (Oryza sativa L. ssp. japonica) Qiufeng in the field. The first three leaves of the mutant were albino with some green. The leaf color became pale green since the fourth leaf and the glume had the same phenomenon as the first three leaves. The measuring data of the pigment content confirmed the visually observed results. It truly had a remarkable changing process in the leaf color in Qiufeng M. Comparison of the main agronomic characters between Qiufeng and Qiufeng M indicated that the neck length and grain weight showed significant difference at the 1% level, and other characters were not different. Genetic analysis showed that the green-revertible albino trait was controlled by a single recessive nucleic gene. Using 209 recessive mutant individuals in the F(2) population derived from the cross Peiai 64S x Qiufeng M, a gene, tentatively named gra((t)), was located between the SSR markers of RM475 and RM2-22 on the long arm of chromosome 2. The genetic distance were 17.3 cM and 2.9 cM respectively.

Fine mapping and candidate gene analysis of a green-revertible albino gene gra(t) in rice.

Green-revertible albino is a novel type of chlorophyll deficiency in rice (Oryza sativa L.), which is helpful for further research in chlorophyll synthesis and chloroplast development to illuminate their molecular mechanism. In the previous study, we had reported a single recessive gene, gra(t), controlling this trait on the long arm of chromosome 2. In this paper, we mapped the gra(t) gene using 1,936 recessive individuals with albino phenotype in the F(2) population derived from the cross between themo-photoperiod-sensitive genic male-sterile (T/PGMS) line Peiai 64S and the spontaneous mutant Qiufeng M. Eventually, it was located to a confined region of 42.4 kb flanked by two microsatellite markers RM2-97 and RM13553. Based on the annotation results of RiceGAAS system, 11 open reading frames (ORFs) were predicted in this region. Among them, ORF6 was the most possible gene related to chloroplast development, which encoded the chloroplast protein synthesis elongation factor Tu in rice. Therefore, we designated it as the candidate gene of gra(t). Sequence analysis indicated that only one base substitution C to T occurred in the coding region, which caused a missense mutation (Thr to Ile) in gra(t) mutant. These results are very valuable for further study on gra(t) gene.

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.

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.