The extraction of DNA is often the most time consuming and laborious step in high-throughput molecular genetic analysis and marker assisted selection （MAS） programs. A simple method for preparation of rice genomic D...The extraction of DNA is often the most time consuming and laborious step in high-throughput molecular genetic analysis and marker assisted selection （MAS） programs. A simple method for preparation of rice genomic DNA was developed. A small amount （1~50 mg） of leaf tissue of rice seedling, 500 pL of extraction buffer, and one steel bead were put into a 2-mL microcentrifuge tube. After vigorously mashing for 2 min, 5 μL of supernatant was directly applied to PCR amplification. Otherwise, the supematant was precipitated with two times volume of ethanol to obtain high quality genomic DNA. This method is simple, rapid, low cost, and reliable for PCR analysis. One person can manipulate as many as 96 samples for PCR in 10 min. It is especially suitable for genotyping of large number of samples.展开更多
Moderate leaf rolling can maintain leaf erectness,improve light transmittance in the population,and improve light energy utilization,thereby increasing rice yield.This study used ethyl methanesulfonate(EMS)to treat Yu...Moderate leaf rolling can maintain leaf erectness,improve light transmittance in the population,and improve light energy utilization,thereby increasing rice yield.This study used ethyl methanesulfonate(EMS)to treat Yunjing 17(YJ17)and obtained a semi-rolled leaf mutant that was named semi-rolled leaf 3(srl3).We found that the rolled-leaf phenotype was due to the aberrant development of bulliform cells and the loss of sclerenchymatous cells.In addition,the shoot and root length of srl3 seedlings differed from the wild type.The srl3 mutant had significantly lower plant height and seed-setting rate but notably greater tiller number,panicle length,and primary branch number per panicle than the wild type.Genetic analysis showed that a single recessive nuclear gene defined the srl3 mutant,and it was precisely located in a 144-kb region between two insertion-deletion(InDel)markers,M8 and M19,on chromosome 2.In this region,no leaf-rollingrelated genes have been reported previously.Thus,the study indicated that SRL3 is a novel leaf-rolling-related gene,and the results laid the foundation for the cloning and functional analysis of the SRL3 gene.展开更多
Photosynthesis occurs mainly in chloroplasts,whose development is regulated by proteins encoded by nuclear genes.Among them,pentapeptide repeat(PPR)proteins participate in organelle RNA editing.Although there are more...Photosynthesis occurs mainly in chloroplasts,whose development is regulated by proteins encoded by nuclear genes.Among them,pentapeptide repeat(PPR)proteins participate in organelle RNA editing.Although there are more than 450 members of the PPR protein family in rice,only a few affect RNA editing in rice chloroplasts.Gene editing technology has created new rice germplasm and mutants,which could be used for rice breeding and gene function study.This study evaluated the functions of OsPPR9 in chloroplast RNA editing in rice.The osppr9 mutants were obtained by CRISPR/Cas9,which showed yellowing leaves and a lethal phenotype,with suppressed expression of genes associated with chloroplast development and accumulation of photosynthetic-related proteins.In addition,loss of OsPPR9 protein function reduces the editing efficiency of rps8-C182,rpoC2-C4106,rps14-C80,and ndhB-C611 RNA editing sites,which affects chloroplast growth and development in rice.Our data showed that OsPPR9 is highly expressed in rice leaves and encodes a DYW-PPR protein localized in chloroplasts.Besides,the OsPPR9 protein was shown to interact with OsMORF2 and OsMORF9.Together,our findings provide insights into the role of the PPR protein in regulating chloroplast development in rice.展开更多
Sheath blight（SB） disease,caused by Rhizoctonia solani Kuhn,is one of the most serious diseases causing rice（Oryza sativa L.） yield loss worldwide.A doubled haploid（DH） population was constructed from a cross be...Sheath blight（SB） disease,caused by Rhizoctonia solani Kuhn,is one of the most serious diseases causing rice（Oryza sativa L.） yield loss worldwide.A doubled haploid（DH） population was constructed from a cross between a japonica variety CJ06 and an indica variety TN1,and to analyze the quantitative trait loci（QTLs） for SB resistance under three different environments（environments 1-3）.Two traits were recorded to evaluate the SB resistance,namely lesion height（LH） and disease rating（DR）.Based on field evaluation of SB resistance and a genetic map constructed with 214 markers,a total of eight QTLs were identified for LH and eight QTLs for DR under three environments,respectively.The QTLs for LH were anchored on chromosomes 1,3,4,5,6,and 8,and explained 4.35-17.53%of the phenotypic variation.The SB resistance allele of qHNLH4 from TN1 decreased LH by 3.08 cm,and contributed to 17.53%of the variation at environment 1.The QTL for LH（qHZaLH8） detected on chromosome 8 in environment 2 explained 16.71%of the variation,and the resistance allele from CJ06 reduced LH by 4.4 cm.Eight QTLs for DR were identified on chromosomes 1,5,6,8,9,11,and 12 under three conditions with the explained variation from 2.0 to 11.27%.The QTL for DR（qHZaDR8）,which explained variation of 11.27%,was located in the same interval as that of qHZaLH8,both QTLs were detected in environment 2.A total of six pairs of digenic epistatic loci for DR were detected in three conditions,but no epistatic locus was observed for LH.In addition,we detected 12 QTLs for plant height（PH） in three environments.None of the PH-QTLs were co-located with the SB-QTLs.The results facilitate our understanding of the genetic basis for SB resistance in rice.展开更多
Field performance of whitebacked planthopper （WBPH）-resistance of four phenotypes was evaluated in Chunjiang 06 （C J-06） / TN1 DH rice lines, which were expressed by different combinations of sucking inhibitory an...Field performance of whitebacked planthopper （WBPH）-resistance of four phenotypes was evaluated in Chunjiang 06 （C J-06） / TN1 DH rice lines, which were expressed by different combinations of sucking inhibitory and ovicidal traits inherited independently from C J-06. WBPH established the highest populations in susceptible DH lines that had neither sucking inhibitorynor ovicidal resistance. Both immigration and subsequent population levels were kept below the damage-causing density in the sucking inhibitory DH lines even under a WBPH outbreak. WBPH could not build up populations in the DH lines having both the sucking inhibitory and ovicidal resistance. Although WBPH immigrated preferentially to non-sucking inhibitory DH lines with ovicidal resistance, subsequent population buildup was significantly suppressed. It was concluded that the differential performance to WBPH-resistance in CJ-06 / TN1 DH lines was primarily due to the sucking inhibitory trait, and complementarity to the ovicidal trait.展开更多
A mutant of panicle differentiation in rice called non-panicle （nop） was discovered in the progeny of a cross between 93-11 and Nipponbare. The mutant exhibits normal plant morphology but has apparently few tillers....A mutant of panicle differentiation in rice called non-panicle （nop） was discovered in the progeny of a cross between 93-11 and Nipponbare. The mutant exhibits normal plant morphology but has apparently few tillers. The most striking change in nop is that its panicle differentiation is blocked, with masses of fluffy bract nodes generate from the positions where rachis branches normally develop in wild-type plants. Genetic analysis suggests that nop is controlled by a single recessive gene, which is temporarily named Nop（t）. Based on its mutant phenotype, Nop（t） represents a key gene controlling the initiation of inflorescence differentiation, By using simple sequence repeat markers and sequence tagged site markers, Nop（t） gene was fine mapped in a 102-kb interval on the long arm of chromosome 6. These results will facilitate the positional cloning and functional studies of the gene.展开更多
基金supported by grants from the Ministry of Agriculture of China for transgenic research(Grant No.2008ZX08009-003)the National Natural Science Foundation of China(Grant Nos.30710103903 and 30771160)the Natural Science Foundation of Zhejiang Province,China(Grant No.R3090023)
文摘The extraction of DNA is often the most time consuming and laborious step in high-throughput molecular genetic analysis and marker assisted selection （MAS） programs. A simple method for preparation of rice genomic DNA was developed. A small amount （1~50 mg） of leaf tissue of rice seedling, 500 pL of extraction buffer, and one steel bead were put into a 2-mL microcentrifuge tube. After vigorously mashing for 2 min, 5 μL of supernatant was directly applied to PCR amplification. Otherwise, the supematant was precipitated with two times volume of ethanol to obtain high quality genomic DNA. This method is simple, rapid, low cost, and reliable for PCR analysis. One person can manipulate as many as 96 samples for PCR in 10 min. It is especially suitable for genotyping of large number of samples.
基金supported by the National Natural Science Foundation of China(32071993 and 91735304)。
文摘Moderate leaf rolling can maintain leaf erectness,improve light transmittance in the population,and improve light energy utilization,thereby increasing rice yield.This study used ethyl methanesulfonate(EMS)to treat Yunjing 17(YJ17)and obtained a semi-rolled leaf mutant that was named semi-rolled leaf 3(srl3).We found that the rolled-leaf phenotype was due to the aberrant development of bulliform cells and the loss of sclerenchymatous cells.In addition,the shoot and root length of srl3 seedlings differed from the wild type.The srl3 mutant had significantly lower plant height and seed-setting rate but notably greater tiller number,panicle length,and primary branch number per panicle than the wild type.Genetic analysis showed that a single recessive nuclear gene defined the srl3 mutant,and it was precisely located in a 144-kb region between two insertion-deletion(InDel)markers,M8 and M19,on chromosome 2.In this region,no leaf-rollingrelated genes have been reported previously.Thus,the study indicated that SRL3 is a novel leaf-rolling-related gene,and the results laid the foundation for the cloning and functional analysis of the SRL3 gene.
基金funded by the Central Public-Interest Scientific Institution Basal Research Fund,China(CPSIBRF-CNRRI-202111 and CPSIBRF-CNRRI-202110)the Agricultural Science and Technology Innovation Program,Chinese Academy of Agricultural Sciences(ASTIP)+1 种基金the Project of State Key Laboratory of Rice Biology,China(2020ZZKT10205)the Key Research and Development Project of China Rice Research Institute(CNRRI-2020-01)。
文摘Photosynthesis occurs mainly in chloroplasts,whose development is regulated by proteins encoded by nuclear genes.Among them,pentapeptide repeat(PPR)proteins participate in organelle RNA editing.Although there are more than 450 members of the PPR protein family in rice,only a few affect RNA editing in rice chloroplasts.Gene editing technology has created new rice germplasm and mutants,which could be used for rice breeding and gene function study.This study evaluated the functions of OsPPR9 in chloroplast RNA editing in rice.The osppr9 mutants were obtained by CRISPR/Cas9,which showed yellowing leaves and a lethal phenotype,with suppressed expression of genes associated with chloroplast development and accumulation of photosynthetic-related proteins.In addition,loss of OsPPR9 protein function reduces the editing efficiency of rps8-C182,rpoC2-C4106,rps14-C80,and ndhB-C611 RNA editing sites,which affects chloroplast growth and development in rice.Our data showed that OsPPR9 is highly expressed in rice leaves and encodes a DYW-PPR protein localized in chloroplasts.Besides,the OsPPR9 protein was shown to interact with OsMORF2 and OsMORF9.Together,our findings provide insights into the role of the PPR protein in regulating chloroplast development in rice.
基金supported by the National Natural Science Foundation of China(31101004,31221004)the National High-Tech R&D Program of China(863 Program,2012AA101201)+2 种基金a fund from Zhejiang Province for public welfare(2014C32013)a special fund for technical innovation team in Zhejiang Province,China(2010R50024)a fund from the Chinese Academy of Agricultural Sciences to the Scientific and Technical Innovation Team
文摘Sheath blight（SB） disease,caused by Rhizoctonia solani Kuhn,is one of the most serious diseases causing rice（Oryza sativa L.） yield loss worldwide.A doubled haploid（DH） population was constructed from a cross between a japonica variety CJ06 and an indica variety TN1,and to analyze the quantitative trait loci（QTLs） for SB resistance under three different environments（environments 1-3）.Two traits were recorded to evaluate the SB resistance,namely lesion height（LH） and disease rating（DR）.Based on field evaluation of SB resistance and a genetic map constructed with 214 markers,a total of eight QTLs were identified for LH and eight QTLs for DR under three environments,respectively.The QTLs for LH were anchored on chromosomes 1,3,4,5,6,and 8,and explained 4.35-17.53%of the phenotypic variation.The SB resistance allele of qHNLH4 from TN1 decreased LH by 3.08 cm,and contributed to 17.53%of the variation at environment 1.The QTL for LH（qHZaLH8） detected on chromosome 8 in environment 2 explained 16.71%of the variation,and the resistance allele from CJ06 reduced LH by 4.4 cm.Eight QTLs for DR were identified on chromosomes 1,5,6,8,9,11,and 12 under three conditions with the explained variation from 2.0 to 11.27%.The QTL for DR（qHZaDR8）,which explained variation of 11.27%,was located in the same interval as that of qHZaLH8,both QTLs were detected in environment 2.A total of six pairs of digenic epistatic loci for DR were detected in three conditions,but no epistatic locus was observed for LH.In addition,we detected 12 QTLs for plant height（PH） in three environments.None of the PH-QTLs were co-located with the SB-QTLs.The results facilitate our understanding of the genetic basis for SB resistance in rice.
基金JIRCAS International Collaborative Project Fund(B33102-331)Grant Project of Zhejiang Province for International Collaboration(2002AA217111)supported this work
文摘Field performance of whitebacked planthopper （WBPH）-resistance of four phenotypes was evaluated in Chunjiang 06 （C J-06） / TN1 DH rice lines, which were expressed by different combinations of sucking inhibitory and ovicidal traits inherited independently from C J-06. WBPH established the highest populations in susceptible DH lines that had neither sucking inhibitorynor ovicidal resistance. Both immigration and subsequent population levels were kept below the damage-causing density in the sucking inhibitory DH lines even under a WBPH outbreak. WBPH could not build up populations in the DH lines having both the sucking inhibitory and ovicidal resistance. Although WBPH immigrated preferentially to non-sucking inhibitory DH lines with ovicidal resistance, subsequent population buildup was significantly suppressed. It was concluded that the differential performance to WBPH-resistance in CJ-06 / TN1 DH lines was primarily due to the sucking inhibitory trait, and complementarity to the ovicidal trait.
基金supported by the grants from the National Natural Science Foundation of China (Grant No.30300196 and No. 30771160)the State Key Basic Research Program of China (Grant No.2007CB10920203)the Research Program of Zhejiang Province,China
文摘A mutant of panicle differentiation in rice called non-panicle （nop） was discovered in the progeny of a cross between 93-11 and Nipponbare. The mutant exhibits normal plant morphology but has apparently few tillers. The most striking change in nop is that its panicle differentiation is blocked, with masses of fluffy bract nodes generate from the positions where rachis branches normally develop in wild-type plants. Genetic analysis suggests that nop is controlled by a single recessive gene, which is temporarily named Nop（t）. Based on its mutant phenotype, Nop（t） represents a key gene controlling the initiation of inflorescence differentiation, By using simple sequence repeat markers and sequence tagged site markers, Nop（t） gene was fine mapped in a 102-kb interval on the long arm of chromosome 6. These results will facilitate the positional cloning and functional studies of the gene.
