We identified the rice floral organ development mutant, termed as open hull and male sterile 1 （ohms1）, from the progeny of the indica restorer line Zhonghui 8015 treated with 60Co γ-ray irradiation. The ohms1 muta...We identified the rice floral organ development mutant, termed as open hull and male sterile 1 （ohms1）, from the progeny of the indica restorer line Zhonghui 8015 treated with 60Co γ-ray irradiation. The ohms1 mutant exhibited an open hull and lemma- and palea-like structure conversion between the anthers and stigma, which resulted in the ohms1 mutant spikelet showing‘tridentate lemma＇. The ohms1 mutant was entirely sterile but had 60%-70% fertile pollen. Genetic analysis and gene mapping showed that ohms1 was controlled by a single recessive gene, and the mutant gene was fine-mapped to a 42-kb interval on the short arm of chromosome 3 between markers KY2 and KY29. Sequence analysis of the four open reading frames in this region revealed that the mutant carried a single nucleotide transformation （A to G） at the last base of the fifth intron, which was likely corresponded to ohms1 phynotype, in an MIKC type MADS-box gene OsMADS1 （LOC_Os03g11614）. Enzyme digestion and cDNA sequencing further indicated that the variable splicing was responsible for the deletion of the sixth exon in ohms1, but no structural changes in the MADS domain or amino acid frame shifts appeared. Additionally, real-time fluorescent quantitative PCR analysis showed that the OsMADS1 expression level decreased significantly in the ohms1 mutant. The expression levels of rice flowering factors and floral glume development-related genes also changed significantly. These results demonstrate that OsMADS1 may play an important role in rice floral organ development, particularly in floral glume development and floret primordium differentiation.展开更多
Male sterile genes and mutants are valuable resources in hybrid seed production for monoclinous crops.High genetic redundancy due to allohexaploidy makes it difficult to obtain the nuclear recessive male sterile mutan...Male sterile genes and mutants are valuable resources in hybrid seed production for monoclinous crops.High genetic redundancy due to allohexaploidy makes it difficult to obtain the nuclear recessive male sterile mutants through spontaneous mutation or chemical or physical mutagenesis methods in wheat.The emerging effective genome editing tool,CRISPR/Cas9 system,makes it possible to achieve simultaneous mutagenesis in multiple homoeoalleles.To improve the genome modification efficiency of the CRISPR/Cas9 system in wheat,we compared four different RNA polymerase(Pol)Ⅲpromoters(TaU3 p,TaU6 p,OsU3 p,and OsU6 p)and three types of sgRNA scaffold in the protoplast system.We show that the TaU3 promoter-driven optimized sgRNA scaffold was most effective.The optimized CRISPR/Cas9 system was used to edit three TaNP1 homoeoalleles,whose orthologs,OsNP1 in rice and ZmIPE1 in maize,encode a putative glucose-methanol-choline oxidoreductase and are required for male sterility.Triple homozygous mutations in TaNP1 genes result in complete male sterility.We further demonstrated that anyone wild-type copy of the three TaNP1 genes is sufficient for maintenance of male fertility.Taken together,this study provides an optimized CRISPR/Cas9 vector for wheat genome editing and a complete male sterile mutant for development of a commercially viable hybrid wheat seed production system.展开更多
With the cDNA suppression subtraction hybridization method, a spikelet-specific cDNA library was constructed that expressed at meiosis stage in rice. A total of 121 cDNA fragments were selected from the library and us...With the cDNA suppression subtraction hybridization method, a spikelet-specific cDNA library was constructed that expressed at meiosis stage in rice. A total of 121 cDNA fragments were selected from the library and used as EST (expressed se-quence tags) markers to detect the polymorphism between Annong N, a normal fertile Indica rice line and Annong S-1, its spontaneous mutant with thermo-sensitive genic male sterility, using the RFLP (restriction fragment length polymorphism) technique. HN57, one of the EST probes, could detect poly-morphism between them. The results of segregation analysis with the F2 population developed from An-nong S-1 and Annong N indicate that HN57 co-seg- regates with the thermo-sensitive genic male-sterility controlled by tms5, the recessive gene in Annong S-1. This marker is located on the 31.2-cM region of the chromosome 2 of RGP (rice genome research pro-gram) genetic map. To further determine the location of tms5, 80 SSR (simple sequence repeat) markers around this region were developed, and 12 of them were polymorphic. And finally, the tms5 was mapped within region of 181 kb by using these new markers.展开更多
THE first cytoplasmic male sterile soybean line was reported by Davis in a U.S. Patent, but no further information has been released since then. Sun et al. developed a cytoplasmic-nuclear male sterile soy-
One sequence tagged site marker Subl-1 and twice submergence stress method were used in selection of submergence tolerant homozygous line from Sub-lBS, a submergence tolerant, bentazon sensitive and photoperiod-sensit...One sequence tagged site marker Subl-1 and twice submergence stress method were used in selection of submergence tolerant homozygous line from Sub-lBS, a submergence tolerant, bentazon sensitive and photoperiod-sensitive and/or thermo-sensitive genic male sterile line that developed by our laboratory. The results revealed that the original Sub-lBS was heterozygous in SublA-1 locus even though it was identical in almost all of agronomical traits and the segregation of SublA-1 was in accordance with Mendelian law based on chi-square test. And then the original Sub-IBS was divided into two groups： one was ofSublA-1 introgression and the other was not; and the two groups were tested by twice submergence stress method. After the first submergence stress that lasted for 12 d, the average plant heights were significant difference at the 1% level between the two groups. After recovery for 10 d, the second submergence stress sustained for 18 d was carried on; and the group with SublA-1 gene was found apparently tolerant than the other group in submergence tolerance.展开更多
基金supported by the National Key Technology R&D Program of China (Grant No.2011BAD35B02)the National Key Transform Program of China (Grant No.2011ZX08001-002)+1 种基金Zhejiang Provincial Natural Science Foundation of China (Grant No.LQ14C130003)the Super Rice Breeding Innovation Team and Rice Heterosis Mechanism Research Innovation Team of the Chinese Academy of Agricultural Sciences Innovation Project, China (Grant No.CAAS-ASTIP2013-CNRRI)
文摘We identified the rice floral organ development mutant, termed as open hull and male sterile 1 （ohms1）, from the progeny of the indica restorer line Zhonghui 8015 treated with 60Co γ-ray irradiation. The ohms1 mutant exhibited an open hull and lemma- and palea-like structure conversion between the anthers and stigma, which resulted in the ohms1 mutant spikelet showing‘tridentate lemma＇. The ohms1 mutant was entirely sterile but had 60%-70% fertile pollen. Genetic analysis and gene mapping showed that ohms1 was controlled by a single recessive gene, and the mutant gene was fine-mapped to a 42-kb interval on the short arm of chromosome 3 between markers KY2 and KY29. Sequence analysis of the four open reading frames in this region revealed that the mutant carried a single nucleotide transformation （A to G） at the last base of the fifth intron, which was likely corresponded to ohms1 phynotype, in an MIKC type MADS-box gene OsMADS1 （LOC_Os03g11614）. Enzyme digestion and cDNA sequencing further indicated that the variable splicing was responsible for the deletion of the sixth exon in ohms1, but no structural changes in the MADS domain or amino acid frame shifts appeared. Additionally, real-time fluorescent quantitative PCR analysis showed that the OsMADS1 expression level decreased significantly in the ohms1 mutant. The expression levels of rice flowering factors and floral glume development-related genes also changed significantly. These results demonstrate that OsMADS1 may play an important role in rice floral organ development, particularly in floral glume development and floret primordium differentiation.
基金supported by grants from the Ministry of Agriculture of China(2016ZX08010001 and 2016ZX08010002)Peking University Institute of Advanced Agricultural Sciences and Beijing Natural Science Foundation(19530290014)。
文摘Male sterile genes and mutants are valuable resources in hybrid seed production for monoclinous crops.High genetic redundancy due to allohexaploidy makes it difficult to obtain the nuclear recessive male sterile mutants through spontaneous mutation or chemical or physical mutagenesis methods in wheat.The emerging effective genome editing tool,CRISPR/Cas9 system,makes it possible to achieve simultaneous mutagenesis in multiple homoeoalleles.To improve the genome modification efficiency of the CRISPR/Cas9 system in wheat,we compared four different RNA polymerase(Pol)Ⅲpromoters(TaU3 p,TaU6 p,OsU3 p,and OsU6 p)and three types of sgRNA scaffold in the protoplast system.We show that the TaU3 promoter-driven optimized sgRNA scaffold was most effective.The optimized CRISPR/Cas9 system was used to edit three TaNP1 homoeoalleles,whose orthologs,OsNP1 in rice and ZmIPE1 in maize,encode a putative glucose-methanol-choline oxidoreductase and are required for male sterility.Triple homozygous mutations in TaNP1 genes result in complete male sterility.We further demonstrated that anyone wild-type copy of the three TaNP1 genes is sufficient for maintenance of male fertility.Taken together,this study provides an optimized CRISPR/Cas9 vector for wheat genome editing and a complete male sterile mutant for development of a commercially viable hybrid wheat seed production system.
文摘With the cDNA suppression subtraction hybridization method, a spikelet-specific cDNA library was constructed that expressed at meiosis stage in rice. A total of 121 cDNA fragments were selected from the library and used as EST (expressed se-quence tags) markers to detect the polymorphism between Annong N, a normal fertile Indica rice line and Annong S-1, its spontaneous mutant with thermo-sensitive genic male sterility, using the RFLP (restriction fragment length polymorphism) technique. HN57, one of the EST probes, could detect poly-morphism between them. The results of segregation analysis with the F2 population developed from An-nong S-1 and Annong N indicate that HN57 co-seg- regates with the thermo-sensitive genic male-sterility controlled by tms5, the recessive gene in Annong S-1. This marker is located on the 31.2-cM region of the chromosome 2 of RGP (rice genome research pro-gram) genetic map. To further determine the location of tms5, 80 SSR (simple sequence repeat) markers around this region were developed, and 12 of them were polymorphic. And finally, the tms5 was mapped within region of 181 kb by using these new markers.
文摘THE first cytoplasmic male sterile soybean line was reported by Davis in a U.S. Patent, but no further information has been released since then. Sun et al. developed a cytoplasmic-nuclear male sterile soy-
基金supported by the Knowledge Innovation Program of the Chinese Academy of Sciences(KSCX2-EW-N-01)
文摘One sequence tagged site marker Subl-1 and twice submergence stress method were used in selection of submergence tolerant homozygous line from Sub-lBS, a submergence tolerant, bentazon sensitive and photoperiod-sensitive and/or thermo-sensitive genic male sterile line that developed by our laboratory. The results revealed that the original Sub-lBS was heterozygous in SublA-1 locus even though it was identical in almost all of agronomical traits and the segregation of SublA-1 was in accordance with Mendelian law based on chi-square test. And then the original Sub-IBS was divided into two groups： one was ofSublA-1 introgression and the other was not; and the two groups were tested by twice submergence stress method. After the first submergence stress that lasted for 12 d, the average plant heights were significant difference at the 1% level between the two groups. After recovery for 10 d, the second submergence stress sustained for 18 d was carried on; and the group with SublA-1 gene was found apparently tolerant than the other group in submergence tolerance.
