To understand the resistance inheritance basis of space-induced rice lines to blast, and to probe mutants' genomic DNA polymorphism compared with ground control by microsatellite markers, three space-induced lines we...To understand the resistance inheritance basis of space-induced rice lines to blast, and to probe mutants' genomic DNA polymorphism compared with ground control by microsatellite markers, three space-induced lines were crossed with a highly susceptible variety LTH, and their F1 and F2 populations were inoculated by two representative blast isolates with broad pathogenicity to analyze their resistance inheritance basis. Meanwhile three mutant lines and the ground control were analyzed by 225 rice SSR (simple sequence repeat) primer pairs selected throughout the 12 chromosomes of whole rice genome, to scan the mutagenesis in genome of the mutant lines. The results indicated the blast-resistant genes harbored in these mutant lines were dominant. It was demonstrated that the resistance of mutant H1 to isolate GD0193 and GD3286 was controlled by a single gene, respectively; while mutants H2 and H3 were controlled by two pairs of major genes against isolate GD3286 and H2 showed complicated genetic mechanism to isolate GD0193. H3's resistance to isolate GD0193 was verified to be controlled by a single gene. According to the results of SSR analysis, three mutant lines showed different mutant rates as compared with the ground control, and the mutant rates also varied. Resistance genes can be induced from rice by space mutation, and different genomic variations were detected in blast-resistant lines.展开更多
Blast resistance and grain quality are major problems in hybrid rice production in China. In this study, two resistance (R) genes, Pi46 and Pita, along with the gene Wxb, which mainly affects rice endosperm amylose ...Blast resistance and grain quality are major problems in hybrid rice production in China. In this study, two resistance (R) genes, Pi46 and Pita, along with the gene Wxb, which mainly affects rice endosperm amylose content (AC), were introgressed into an elite indica restoring line, R8166, which has little blast resistance and poor grain quality through marker-assisted selection (MAS). Eight improved lines were found to have recurrent genome recovery ratios ranging from 88.68 to 96.23%. Two improved lines, R163 and R167, were selected for subsequent studies. R167, which has the highest recovery ratio (96.23%), showed no significant differences in multiple agronomic traits. In contrast, R163 with the lowest recovery ratio (88.68%) exhibited significant differences in heading date and yield per plant compared with the recurrent parent. At two developmental stages, R163 and R167 had greatly enhanced resistance to blast over the recurrent parent. Similar trends were also observed for agronomic traits and blast resistance in R163- and R167-derived hybrids when compared with the counterparts from R8166. In addition, R163, R167, and their derived hybrids significantly improved the grain quality traits, including amylose content (AC), gel consistency (GC), chalky grain rate (CGR), and degree of endosperm chalkiness (DEC). It confirmed the success of efficiently developing elite restoring lines using MAS in this study.展开更多
In the rice cytoplasmic-genetic male sterility (CMS) system, the combination of a CMS line, maintainer line and restorer line carrying the restorer gene to restore fertility, is indispensable for the development of ...In the rice cytoplasmic-genetic male sterility (CMS) system, the combination of a CMS line, maintainer line and restorer line carrying the restorer gene to restore fertility, is indispensable for the development of hybrids. However, the process of screening for the trait of fertility restoration is laborious and time-consuming. In the present study, we analyzed the nucleotide sequence of the Rf4 gene, which is the major locus controlling fertility restoration, to identify allele-specific variation. A single nucleotide polymorphism (SNP) A/C at +474 in the coding sequence (CDS) was found to be capable of strictly distinguishing groups of alleles Rf4 (A) and rf4 (C). Using KASP genotyping, this valuable SNP was converted to an allele-specific PCR marker. We evaluated and validated the marker among three-line parents with different backgrounds, and the results revealed a complete correlation between SNP alleles and the fertility restoration phenotype. Molecular screening was subsequently carried out for the presence of alleles of Rf4 and Rf3 among 328 diverse rice cultivars with worldwide distribution. The results demonstrate that this SNP marker could be the optimal choice for the molecular identification of potential restorers.展开更多
Utilization of R(resistance) genes to develop resistant cultivars is an effective strategy to combat against rice blast disease. In this study, R genes Pi46 and Pita in a resistant accession H4 were introgressed int...Utilization of R(resistance) genes to develop resistant cultivars is an effective strategy to combat against rice blast disease. In this study, R genes Pi46 and Pita in a resistant accession H4 were introgressed into an elite restorer line Hang-Hui-179(HH179) using the marker-assisted backcross breeding(MABB) procedure. As a result, three improved lines(e.g., R1791 carrying Pi46 alone, R1792 carrying Pita alone and R1793 carrying both Pi46 and Pita) were developed. The three improved lines had significant genetic similarities with the recurrent parent HH179. Thus, they and HH179 could be recognized as near isogenic lines(NILs). The resistance spectrum of the three improved lines, which was tested at seedling stage, reached 91.1, 64.7 and 97.1%, respectively. This was markedly broader than that of HH179(23.5%). Interestingly, R1793 showed resistance to panicle blast but neither R1791 nor R1792 exhibited resistance at two natural blast nurseries. The results implied that the stacking of Pi46 and Pita resulted in enhanced resistance, which was unachievable by either R gene alone. Further comparison indicated that the three improved lines were similar to HH179 in multiple agronomic traits; including plant height, tillers per plant, panicle length, spikelet fertility, and 1 000-grain weight. Thus, the three improved lines with different R genes can be used as new sources of resistance for developing variety. There is a complementary effect between the two R genes Pi46 and Pita.展开更多
Genetic diversity of 299 inbred indica rice varieties, including 33 introduced varieties, applied in Guangdong Province of China were assessed using 20 ILP (intron length polymorphism) and 34 SSR (simple sequence r...Genetic diversity of 299 inbred indica rice varieties, including 33 introduced varieties, applied in Guangdong Province of China were assessed using 20 ILP (intron length polymorphism) and 34 SSR (simple sequence repeat) markers. Totally, 154 loci were screened for the 299 varieties, with the average number of alleles (Na), rare alleles (Nr), and polymorphism information content (PIC) scored at 3.4, 0.7 and 0.32, respectively. The Nei's genetic distance (GD) was estimated ranging from 0 to 0.7529 with an average of 0.4797. There was no significant difference of Na, Nr, PIC or GDs between the introduced and local varieties. Neighbor-joining (N J) analysis showed that the 299 varieties failed into three main distinct groups, and the 33 introduced varieties were distributed over all the groups or subgroups. Model-based cluster analysis demonstrated that only 73 (24.4%) of the 299 varieties and 7 (21.2%) of the 33 introduced varieties could be distinctly classified into the three groups. Analysis of molecular variance showed that within the groups divided by NJ analysis, the genetic variations revealed by ILP, SSR and these two combined were 7.7%, 5.6% and 6.6%, and within the groups divided by region (Guangdong local and the introduced varieties), the genetic variables were 2.1%, 4.6%, 5.4%, respectively. These results suggested that the genetic diversity of the 299 inbred rice varieties in Guangdong Province was low, simultaneously relationship among varieties was poor and close in all kind of groups. Hence, it is very necessary to extend the genetic diversity during the breeding and selection practical procedure.展开更多
基金supported by the National 863 Program of China (2002AA2410111)the Natural Science Foundation of Guangdong Province,China (031994)the Special Project of Key Science and Technology of Guangdong Province,China (2004A20107001)
文摘To understand the resistance inheritance basis of space-induced rice lines to blast, and to probe mutants' genomic DNA polymorphism compared with ground control by microsatellite markers, three space-induced lines were crossed with a highly susceptible variety LTH, and their F1 and F2 populations were inoculated by two representative blast isolates with broad pathogenicity to analyze their resistance inheritance basis. Meanwhile three mutant lines and the ground control were analyzed by 225 rice SSR (simple sequence repeat) primer pairs selected throughout the 12 chromosomes of whole rice genome, to scan the mutagenesis in genome of the mutant lines. The results indicated the blast-resistant genes harbored in these mutant lines were dominant. It was demonstrated that the resistance of mutant H1 to isolate GD0193 and GD3286 was controlled by a single gene, respectively; while mutants H2 and H3 were controlled by two pairs of major genes against isolate GD3286 and H2 showed complicated genetic mechanism to isolate GD0193. H3's resistance to isolate GD0193 was verified to be controlled by a single gene. According to the results of SSR analysis, three mutant lines showed different mutant rates as compared with the ground control, and the mutant rates also varied. Resistance genes can be induced from rice by space mutation, and different genomic variations were detected in blast-resistant lines.
基金supported by the grant from the State Scholarship Fund of China (20153069)the National Key R&D Program of China (2016YFD0101100)the earmarked fund for China Agriculture Research System (CARS-01-12)
文摘Blast resistance and grain quality are major problems in hybrid rice production in China. In this study, two resistance (R) genes, Pi46 and Pita, along with the gene Wxb, which mainly affects rice endosperm amylose content (AC), were introgressed into an elite indica restoring line, R8166, which has little blast resistance and poor grain quality through marker-assisted selection (MAS). Eight improved lines were found to have recurrent genome recovery ratios ranging from 88.68 to 96.23%. Two improved lines, R163 and R167, were selected for subsequent studies. R167, which has the highest recovery ratio (96.23%), showed no significant differences in multiple agronomic traits. In contrast, R163 with the lowest recovery ratio (88.68%) exhibited significant differences in heading date and yield per plant compared with the recurrent parent. At two developmental stages, R163 and R167 had greatly enhanced resistance to blast over the recurrent parent. Similar trends were also observed for agronomic traits and blast resistance in R163- and R167-derived hybrids when compared with the counterparts from R8166. In addition, R163, R167, and their derived hybrids significantly improved the grain quality traits, including amylose content (AC), gel consistency (GC), chalky grain rate (CGR), and degree of endosperm chalkiness (DEC). It confirmed the success of efficiently developing elite restoring lines using MAS in this study.
基金provided in part by a grant from the the National Key Research and Development Program of China (2016YFD0102102)the Science and Technology Project of Guangdong Province,China (2015B020231011)the China Agriculture Research System (CARS-01-12)
文摘In the rice cytoplasmic-genetic male sterility (CMS) system, the combination of a CMS line, maintainer line and restorer line carrying the restorer gene to restore fertility, is indispensable for the development of hybrids. However, the process of screening for the trait of fertility restoration is laborious and time-consuming. In the present study, we analyzed the nucleotide sequence of the Rf4 gene, which is the major locus controlling fertility restoration, to identify allele-specific variation. A single nucleotide polymorphism (SNP) A/C at +474 in the coding sequence (CDS) was found to be capable of strictly distinguishing groups of alleles Rf4 (A) and rf4 (C). Using KASP genotyping, this valuable SNP was converted to an allele-specific PCR marker. We evaluated and validated the marker among three-line parents with different backgrounds, and the results revealed a complete correlation between SNP alleles and the fertility restoration phenotype. Molecular screening was subsequently carried out for the presence of alleles of Rf4 and Rf3 among 328 diverse rice cultivars with worldwide distribution. The results demonstrate that this SNP marker could be the optimal choice for the molecular identification of potential restorers.
基金supported by the grant from the Science and Technology Planning Project of Guangdong Province, China (2015A020209142)by the earmarked fund for the Modern Agro-industry Technology Research System,China (CARS-01-12)
文摘Utilization of R(resistance) genes to develop resistant cultivars is an effective strategy to combat against rice blast disease. In this study, R genes Pi46 and Pita in a resistant accession H4 were introgressed into an elite restorer line Hang-Hui-179(HH179) using the marker-assisted backcross breeding(MABB) procedure. As a result, three improved lines(e.g., R1791 carrying Pi46 alone, R1792 carrying Pita alone and R1793 carrying both Pi46 and Pita) were developed. The three improved lines had significant genetic similarities with the recurrent parent HH179. Thus, they and HH179 could be recognized as near isogenic lines(NILs). The resistance spectrum of the three improved lines, which was tested at seedling stage, reached 91.1, 64.7 and 97.1%, respectively. This was markedly broader than that of HH179(23.5%). Interestingly, R1793 showed resistance to panicle blast but neither R1791 nor R1792 exhibited resistance at two natural blast nurseries. The results implied that the stacking of Pi46 and Pita resulted in enhanced resistance, which was unachievable by either R gene alone. Further comparison indicated that the three improved lines were similar to HH179 in multiple agronomic traits; including plant height, tillers per plant, panicle length, spikelet fertility, and 1 000-grain weight. Thus, the three improved lines with different R genes can be used as new sources of resistance for developing variety. There is a complementary effect between the two R genes Pi46 and Pita.
基金supported by the Guangdong Natural Science Foundation of China (Grant No. S2012040007829)the National High Technology Research and Development Program of China (Grant No. 2012AA101201)
文摘Genetic diversity of 299 inbred indica rice varieties, including 33 introduced varieties, applied in Guangdong Province of China were assessed using 20 ILP (intron length polymorphism) and 34 SSR (simple sequence repeat) markers. Totally, 154 loci were screened for the 299 varieties, with the average number of alleles (Na), rare alleles (Nr), and polymorphism information content (PIC) scored at 3.4, 0.7 and 0.32, respectively. The Nei's genetic distance (GD) was estimated ranging from 0 to 0.7529 with an average of 0.4797. There was no significant difference of Na, Nr, PIC or GDs between the introduced and local varieties. Neighbor-joining (N J) analysis showed that the 299 varieties failed into three main distinct groups, and the 33 introduced varieties were distributed over all the groups or subgroups. Model-based cluster analysis demonstrated that only 73 (24.4%) of the 299 varieties and 7 (21.2%) of the 33 introduced varieties could be distinctly classified into the three groups. Analysis of molecular variance showed that within the groups divided by NJ analysis, the genetic variations revealed by ILP, SSR and these two combined were 7.7%, 5.6% and 6.6%, and within the groups divided by region (Guangdong local and the introduced varieties), the genetic variables were 2.1%, 4.6%, 5.4%, respectively. These results suggested that the genetic diversity of the 299 inbred rice varieties in Guangdong Province was low, simultaneously relationship among varieties was poor and close in all kind of groups. Hence, it is very necessary to extend the genetic diversity during the breeding and selection practical procedure.