Reduced plant height is one of the most important traits related to lodging resistance and crop yield. The use of reduced height genes has been one of the main features in breeding modern high-yielding wheat varieties...Reduced plant height is one of the most important traits related to lodging resistance and crop yield. The use of reduced height genes has been one of the main features in breeding modern high-yielding wheat varieties with less lodging. A spontaneous dwarf mutant DD399 was identified in a high yielding, gibberellic acid(GA)-insensitive, lodging-resistant variety Nongda 399(ND399). Significant differences in upper internode lengths between mutant DD399 and wild type ND399 were caused by reduced cell elongation. The plant height of ND399 × DD399 F_(1) hybrids was intermediate between the parents, indicating incomplete dominance or a dose–response effect of a reduced height gene. Plant height showed continuous distribution in the F_(2) population, and segregation distortion was observed among the 2292 F_(2:3) progenies. The reduced height mutation was characterized by Illumina 90 K iSelect SNP genotyping and bulked segregant RNA-Seq(BSR-Seq) analysis of the segregating population. A concentrated cluster of polymorphic SNPs associated with the reduced height phenotype was detected in the distal region of chromosome arm 2 BL. Co-segregation of reduced height phenotype with the clustered markers revealed a 36 Mb terminal deletion of chromosome 2 BL in mutant DD399.展开更多
Six pairs of tall and dwarf near-isogenic lines derived from a dominant semi-dwarf mutant (Y98149) were selected to study height expression and sensitivity to gibberellic acid (GA3). The lengths of the 4-5th inter...Six pairs of tall and dwarf near-isogenic lines derived from a dominant semi-dwarf mutant (Y98149) were selected to study height expression and sensitivity to gibberellic acid (GA3). The lengths of the 4-5th internode, the 3rd, 2nd, 1st internodes from the top and the panicle length in the six dwarf near isogenic lines were 97.2%, 53.3%, 65.1%, 61.9% and 94.7% of those in the six tall ones, respectively, indicating that the dominant semi-dwarfing gene significantly inhibited the internode elongation. Moreover, Y98149 (mutant type) was more sensitive to GA3 than Y98148 (wild type), and had a lower GA3 concentration in plant, about 78% of Y98148.展开更多
Many genetic loci for wheat plant height(PH) have been reported, and 26 dwarfing genes have been catalogued. To identify major and stable genetic loci for PH, here we thoroughly summarized these functionally or geneti...Many genetic loci for wheat plant height(PH) have been reported, and 26 dwarfing genes have been catalogued. To identify major and stable genetic loci for PH, here we thoroughly summarized these functionally or genetic verified dwarfing loci from QTL linkage analysis and genome-wide association study published from 2003 to 2022. A total of 332 QTL, 270 GWAS loci and 83 genes for PH were integrated onto chromosomes according to their locations in the IWGSC RefSeq v2.1 and 65 QTL-rich clusters(QRC) were defined. Candidate genes in each QRC were predicted based on IWGSC Annotation v2.1 and the information on functional validation of homologous genes in other species. A total of 38 candidate genes were predicted for 65 QRC including three GA2ox genes in QRC-4B-IV, QRC-5A-VIII and QRC-6A-II(Rht24) as well as GA 20-oxidase 2(TaSD1-3A) in QRC-3A-IV. These outcomes lay concrete foundations for mapbased cloning of wheat dwarfing genes and application in breeding.展开更多
In the winter of 1997, a semi-dwarf mutant was found inthe F6 population of M9056/ R8018 xuan in HainanProvince. In the spring of 1998, the seeds were sown inHefei, Anhui Province and the plant height of the popula-ti...In the winter of 1997, a semi-dwarf mutant was found inthe F6 population of M9056/ R8018 xuan in HainanProvince. In the spring of 1998, the seeds were sown inHefei, Anhui Province and the plant height of the popula-tion was measured at maturity. Results showed that29. 4% of the plants had the same height as the high par-ent had, and 70. 6% remained semi-dwarf. In the winterof 1998, the reciprocal crosses between the remained se-mi-dwarf line and the two wild-type parental lines were展开更多
Plant height is an important trait related to yield potential and plant architecture. A suitable plant height plays a crucial role in improvement of rice yield and lodging resistance. In this study, we found that the ...Plant height is an important trait related to yield potential and plant architecture. A suitable plant height plays a crucial role in improvement of rice yield and lodging resistance. In this study, we found that the traditional upland landrace 'Kaowenghan' (KWH) showed a special semi-dwarf phenotype. To identify the semi-dwarf gene from KWH, we raised BC2F4 semi-dwarf introgression lines (IL) by hybridization of the japonica rice cultivar 'Dianjingyoul' (DJY1) and KWH in a DJY1 background. The plant height of the homozygous semi-dwarf IL (IL-87) was significantly reduced compared with that of DJY1. The phenotype of the F1 progeny of the semi-dwarf IL-87 and DJY1 showed that the semi-dwarf phenotype was semi- dominant. QTL mapping indicated that the semi-dwarf phenotype was controlled by a major QTL qDH1 and was localized between the markers RM6696 and RM12047 on chromosome 1. We also developed near-isogenic lines (NIL) from the BC3F3 population, and found that the yield of homozygous NIL (NIL-2) was not significantly different compared to DJY1. Breeding value evaluation through investigation of the plant height of the progeny of NIL (NIL-2) and cultivars from different genetic background indicate that the novel semi-dwarf gene shows potential as a genetic resource for rice breeding.展开更多
The Green Revolution gene sd1 has been used extensively in modern rice breeding,especially in indica cultivars.However,elite sd1 alleles and related germplasm resources used for japonica rice breeding have not been id...The Green Revolution gene sd1 has been used extensively in modern rice breeding,especially in indica cultivars.However,elite sd1 alleles and related germplasm resources used for japonica rice breeding have not been identified,and extensive efforts are needed for japonica rice breeding to obtain new dwarfing sources.Data from MBKbase-Rice revealed seven sd1 haplotypes in indica and four in japonica rice.Two new sd1 alleles were identified in indica rice.In 295 japonica accessions from northeast Asia,except for the weak functional allele SD1-EQ,sd1-r was the major allele,reducing plant height in comparison with SD1-EQ.Japonica germplasm resources carrying reported sd1 alleles were identified by genotype searching and further verified by literature search,genealogical analysis,and d Caps markers.Pedigrees and geographic distribution showed that sd1-r is an excellent allele widely used in northern China and Tohoku in Japan,and sd1-j is commonly used in east China and Kyushu in Japan.Dongnong-and Xiushui-series cultivars carrying sd1-r and sd1-j,respectively,are essential branches of the backbone parents of Chinese japonica rice,Akihikari and Ce21,with the largest number of descendants and derived generations.In semi-dwarf japonica rice breeding,sd1-d was introgressed into Daohuaxiang 2(DHX2).Dwarf and semi-dwarf lines carrying sd1-d were selected and designated as 1279 and 1280,respectively,after withstanding typhoon-induced strong winds and heavy rains in 2020,and are anticipated to become useful intermediate materials for future genetic research and breeding.This work will facilitate the introduction,parental selection,and marker-assisted breeding,and provide a material basis for the next step in identifying favorable genes that selected together with the sd1 alleles in japonica backbone parents.展开更多
A split-split-plot design was used to evaluate the effects of sowing dates and sowing rates on three winter rape cultivars,including‘PR45D03’,a semi-dwarf hybrid,‘PR46W31’,a traditional hybrid,and‘Californium’,a...A split-split-plot design was used to evaluate the effects of sowing dates and sowing rates on three winter rape cultivars,including‘PR45D03’,a semi-dwarf hybrid,‘PR46W31’,a traditional hybrid,and‘Californium’,an open-pollinated cultivar.August 25 was the optimal sowing date for maximizing protein and oil yields across all three cultivars.Of the cultivars,the traditional hybrid,‘PR46W31’,produced the highest protein and oil yields on that date.The yields of the semi-dwarf hybrid,‘PR45D03’,were greater than those of the open-pollinated cultivar,‘Californium’,when these were sown later than the optimal date.Protein and oil yields did not differ significantly among different seeding densities.展开更多
A mechanized direct seeding of rice with less labor and water usage,has been widely adopted.However,this approach requires varieties that exhibit uniform seedling emergence.Mesocotyl elongation(ME)offers the main driv...A mechanized direct seeding of rice with less labor and water usage,has been widely adopted.However,this approach requires varieties that exhibit uniform seedling emergence.Mesocotyl elongation(ME)offers the main drive of fast emergence of rice seedlings from soils;nevertheless,its genetic basis remains unknown.Here,we identify a major rice quantitative trait locus Mesocotyl Elongation1(qME1),an allele of the Green Revolution gene Semi-Dwarf1(SD1),encoding GA20-oxidase for gibberellin(GA)biosynthesis.ME1 expression is strongly induced by soil depth and ethylene.When rice grains are direct-seeded in soils,the ethylene core signaling factor OsEIL1 directly promotes ME1 transcription,accelerating bioactive GA biosynthesis.The GAs further degrade the DELLA protein SLENDER RICE 1(SLR1),alleviating its inhibition of rice PHYTOCHROME-INTERACTING FACTOR-LIKE13(OsPIL13)to activate the downstream expansion gene OsEXPA4 and ultimately promote rice seedling ME and emergence.The ancient traits of long mesocotyl and strong emergence ability in wild rice and landrace were gradually lost in company with the Green Revolution dwarf breeding process,and an elite ME1-R allele(D349H)is found in some modern Geng varieties(long mesocotyl lengths)in northern China,which can be used in the direct seeding and dwarf breeding of Geng varieties.Furthermore,the ectopic and high expression of ME1 driven by mesocotyl-specific promoters resulted in rice plants that could be direct-seeded without obvious plant architecture or yield penalties.Collectively,we reveal the molecular mechanism of rice ME,and provide useful information for breeding new Green Revolution varieties with long mesocotyl suitable for direct-seeding practice.展开更多
Application and functional study of dwarf and semi-dwarf genes are of great importance to both crop breeding and molecular biology. A new semi-dwarf gene, sd-t(t), non-allelic to sd-1,had been identified in an indica ...Application and functional study of dwarf and semi-dwarf genes are of great importance to both crop breeding and molecular biology. A new semi-dwarf gene, sd-t(t), non-allelic to sd-1,had been identified in an indica rice variety, Aitaiyin 2. In this study the gene was genetically mapped by using an F2 population, which consisted of 474 individuals developed from a cross between Aitaiyin 2 and B30. The sd-t(t) gene was located between the RFLP markers R514 and R1408B with a distance of 1.1 cM to R514, and 4.5 cM to R1408B on chromosome 4. A physical contig covering the sd-t(t) mapping region was further constructed by screening a BAC library with R514 and R1408B as probes, and the physical distance between R514 and R1408B was estimated at approximately 147 kb. This result will facilitate map-based cloning of the sd-t(t) gene.展开更多
Bread wheat(Triticum aestivum L.,AABBDD,2 n=6 x=42),which accounts for most of the cultivated wheat crop worldwide,is a typical allohexaploid with a genome derived from three diploid wild ancestors.Bread wheat arose a...Bread wheat(Triticum aestivum L.,AABBDD,2 n=6 x=42),which accounts for most of the cultivated wheat crop worldwide,is a typical allohexaploid with a genome derived from three diploid wild ancestors.Bread wheat arose and evolved via two sequential allopolyploidization events and was further polished through multiple steps of domestication.Today,cultivated allohexaploid bread wheat has numerous advantageous traits,including adaptive plasticity,favorable yield traits,and extended end-use quality,which have enabled its cultivation well beyond the ranges of its tetraploid and diploid progenitors to become a global staple food crop.In the past decade,rapid advances in wheat genomic research have considerably accelerated our understanding of the bases for the shaping of complex agronomic traits in this polyploid crop.Here,we summarize recent advances in characterizing major genetic factors underlying the origin,evolution,and improvement of polyploid wheats.We end with a brief discussion of the future prospects for the design of gene cloning strategies and modern wheat breeding.展开更多
Although Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/CRISPR-associated 9(Cas9)system has been widely used for basic research in model plants,its application for applied breeding in crops has face...Although Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/CRISPR-associated 9(Cas9)system has been widely used for basic research in model plants,its application for applied breeding in crops has faced strong regulatory obstacles,due mainly to a poor understanding of the authentic output of this system,particularly in higher generations.In this study,different from any previous studies,we investigated in detail the molecular characteristics and production performance of CRISPR/Cas9-generated SD1(semi-dwarf 1)mutants from T2 to T4 generations,of which the selection of T1 and T2 was done only by visual phenotyping for semidwarf plants.Our data revealed not only on-and off-target mutations with small or lager indels but also exogenous elements in T2 plants.All indel mutants passed stably to T3 or T4 without additional modifications independent on the presence of Cas9,while some lines displayed unexpected hereditary patterns of Cas9 or some exogenous elements.In addition,effects of various SD1 alleles on rice height and yield differed depending on genetic backgrounds.Taken together,our data showed that the CRISPR/Cas9 system is effective in producing homozygous mutants for functional analysis,but it may be not as precise as expected in rice,and that early and accurate molecular characterization and screening must be carried out for generations before transitioning of the CRISPR/Cas9 system from laboratory to field.展开更多
基金supported by the National Key Research and Development Program of China(2016YFD0100302)the Science and Technology Service Network Initiative of Chinese Academy of Sciences(KFJ-STS-ZDTP-024)。
文摘Reduced plant height is one of the most important traits related to lodging resistance and crop yield. The use of reduced height genes has been one of the main features in breeding modern high-yielding wheat varieties with less lodging. A spontaneous dwarf mutant DD399 was identified in a high yielding, gibberellic acid(GA)-insensitive, lodging-resistant variety Nongda 399(ND399). Significant differences in upper internode lengths between mutant DD399 and wild type ND399 were caused by reduced cell elongation. The plant height of ND399 × DD399 F_(1) hybrids was intermediate between the parents, indicating incomplete dominance or a dose–response effect of a reduced height gene. Plant height showed continuous distribution in the F_(2) population, and segregation distortion was observed among the 2292 F_(2:3) progenies. The reduced height mutation was characterized by Illumina 90 K iSelect SNP genotyping and bulked segregant RNA-Seq(BSR-Seq) analysis of the segregating population. A concentrated cluster of polymorphic SNPs associated with the reduced height phenotype was detected in the distal region of chromosome arm 2 BL. Co-segregation of reduced height phenotype with the clustered markers revealed a 36 Mb terminal deletion of chromosome 2 BL in mutant DD399.
基金This work was supported by the grants of National Natural Science Foundation of China(No.3037863)Natural Science Foundation of Anhui Province,China(No.01041103).
文摘Six pairs of tall and dwarf near-isogenic lines derived from a dominant semi-dwarf mutant (Y98149) were selected to study height expression and sensitivity to gibberellic acid (GA3). The lengths of the 4-5th internode, the 3rd, 2nd, 1st internodes from the top and the panicle length in the six dwarf near isogenic lines were 97.2%, 53.3%, 65.1%, 61.9% and 94.7% of those in the six tall ones, respectively, indicating that the dominant semi-dwarfing gene significantly inhibited the internode elongation. Moreover, Y98149 (mutant type) was more sensitive to GA3 than Y98148 (wild type), and had a lower GA3 concentration in plant, about 78% of Y98148.
基金funded by the National Natural Science Foundation of China (32101733)Shandong Provincial Natural Science Foundation (ZR202103020229)+1 种基金the High-Level Talents Project of Qingdao Agricultural University (663/1122023)National Natural Science Foundation of China Regional Innovation and Development Joint Fund Project (U22A20457)。
文摘Many genetic loci for wheat plant height(PH) have been reported, and 26 dwarfing genes have been catalogued. To identify major and stable genetic loci for PH, here we thoroughly summarized these functionally or genetic verified dwarfing loci from QTL linkage analysis and genome-wide association study published from 2003 to 2022. A total of 332 QTL, 270 GWAS loci and 83 genes for PH were integrated onto chromosomes according to their locations in the IWGSC RefSeq v2.1 and 65 QTL-rich clusters(QRC) were defined. Candidate genes in each QRC were predicted based on IWGSC Annotation v2.1 and the information on functional validation of homologous genes in other species. A total of 38 candidate genes were predicted for 65 QRC including three GA2ox genes in QRC-4B-IV, QRC-5A-VIII and QRC-6A-II(Rht24) as well as GA 20-oxidase 2(TaSD1-3A) in QRC-3A-IV. These outcomes lay concrete foundations for mapbased cloning of wheat dwarfing genes and application in breeding.
文摘In the winter of 1997, a semi-dwarf mutant was found inthe F6 population of M9056/ R8018 xuan in HainanProvince. In the spring of 1998, the seeds were sown inHefei, Anhui Province and the plant height of the popula-tion was measured at maturity. Results showed that29. 4% of the plants had the same height as the high par-ent had, and 70. 6% remained semi-dwarf. In the winterof 1998, the reciprocal crosses between the remained se-mi-dwarf line and the two wild-type parental lines were
基金funded by grants from National Natural Science Foundation of China(31360330)Chinese Academy of Science(XDA08020203)
文摘Plant height is an important trait related to yield potential and plant architecture. A suitable plant height plays a crucial role in improvement of rice yield and lodging resistance. In this study, we found that the traditional upland landrace 'Kaowenghan' (KWH) showed a special semi-dwarf phenotype. To identify the semi-dwarf gene from KWH, we raised BC2F4 semi-dwarf introgression lines (IL) by hybridization of the japonica rice cultivar 'Dianjingyoul' (DJY1) and KWH in a DJY1 background. The plant height of the homozygous semi-dwarf IL (IL-87) was significantly reduced compared with that of DJY1. The phenotype of the F1 progeny of the semi-dwarf IL-87 and DJY1 showed that the semi-dwarf phenotype was semi- dominant. QTL mapping indicated that the semi-dwarf phenotype was controlled by a major QTL qDH1 and was localized between the markers RM6696 and RM12047 on chromosome 1. We also developed near-isogenic lines (NIL) from the BC3F3 population, and found that the yield of homozygous NIL (NIL-2) was not significantly different compared to DJY1. Breeding value evaluation through investigation of the plant height of the progeny of NIL (NIL-2) and cultivars from different genetic background indicate that the novel semi-dwarf gene shows potential as a genetic resource for rice breeding.
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA24020301)Young Scientists Fund(CN)(31900423)+1 种基金Excellent Youth Foundation for Heilongjiang Scientific Committee(JC2017009)Cooperative Innovation Extension System of Rice Modern Agricultural Industrial Technology in Heilongjiang province。
文摘The Green Revolution gene sd1 has been used extensively in modern rice breeding,especially in indica cultivars.However,elite sd1 alleles and related germplasm resources used for japonica rice breeding have not been identified,and extensive efforts are needed for japonica rice breeding to obtain new dwarfing sources.Data from MBKbase-Rice revealed seven sd1 haplotypes in indica and four in japonica rice.Two new sd1 alleles were identified in indica rice.In 295 japonica accessions from northeast Asia,except for the weak functional allele SD1-EQ,sd1-r was the major allele,reducing plant height in comparison with SD1-EQ.Japonica germplasm resources carrying reported sd1 alleles were identified by genotype searching and further verified by literature search,genealogical analysis,and d Caps markers.Pedigrees and geographic distribution showed that sd1-r is an excellent allele widely used in northern China and Tohoku in Japan,and sd1-j is commonly used in east China and Kyushu in Japan.Dongnong-and Xiushui-series cultivars carrying sd1-r and sd1-j,respectively,are essential branches of the backbone parents of Chinese japonica rice,Akihikari and Ce21,with the largest number of descendants and derived generations.In semi-dwarf japonica rice breeding,sd1-d was introgressed into Daohuaxiang 2(DHX2).Dwarf and semi-dwarf lines carrying sd1-d were selected and designated as 1279 and 1280,respectively,after withstanding typhoon-induced strong winds and heavy rains in 2020,and are anticipated to become useful intermediate materials for future genetic research and breeding.This work will facilitate the introduction,parental selection,and marker-assisted breeding,and provide a material basis for the next step in identifying favorable genes that selected together with the sd1 alleles in japonica backbone parents.
文摘A split-split-plot design was used to evaluate the effects of sowing dates and sowing rates on three winter rape cultivars,including‘PR45D03’,a semi-dwarf hybrid,‘PR46W31’,a traditional hybrid,and‘Californium’,an open-pollinated cultivar.August 25 was the optimal sowing date for maximizing protein and oil yields across all three cultivars.Of the cultivars,the traditional hybrid,‘PR46W31’,produced the highest protein and oil yields on that date.The yields of the semi-dwarf hybrid,‘PR45D03’,were greater than those of the open-pollinated cultivar,‘Californium’,when these were sown later than the optimal date.Protein and oil yields did not differ significantly among different seeding densities.
基金supported by the National Natural Science Foundation of China(32188102 and 32101763)Zhejiang Provincial Science and Technology Project(2020R51007)+1 种基金the Key Research and Development Program of Zhejiang province(2022C02011)the Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-CACB-202402).
文摘A mechanized direct seeding of rice with less labor and water usage,has been widely adopted.However,this approach requires varieties that exhibit uniform seedling emergence.Mesocotyl elongation(ME)offers the main drive of fast emergence of rice seedlings from soils;nevertheless,its genetic basis remains unknown.Here,we identify a major rice quantitative trait locus Mesocotyl Elongation1(qME1),an allele of the Green Revolution gene Semi-Dwarf1(SD1),encoding GA20-oxidase for gibberellin(GA)biosynthesis.ME1 expression is strongly induced by soil depth and ethylene.When rice grains are direct-seeded in soils,the ethylene core signaling factor OsEIL1 directly promotes ME1 transcription,accelerating bioactive GA biosynthesis.The GAs further degrade the DELLA protein SLENDER RICE 1(SLR1),alleviating its inhibition of rice PHYTOCHROME-INTERACTING FACTOR-LIKE13(OsPIL13)to activate the downstream expansion gene OsEXPA4 and ultimately promote rice seedling ME and emergence.The ancient traits of long mesocotyl and strong emergence ability in wild rice and landrace were gradually lost in company with the Green Revolution dwarf breeding process,and an elite ME1-R allele(D349H)is found in some modern Geng varieties(long mesocotyl lengths)in northern China,which can be used in the direct seeding and dwarf breeding of Geng varieties.Furthermore,the ectopic and high expression of ME1 driven by mesocotyl-specific promoters resulted in rice plants that could be direct-seeded without obvious plant architecture or yield penalties.Collectively,we reveal the molecular mechanism of rice ME,and provide useful information for breeding new Green Revolution varieties with long mesocotyl suitable for direct-seeding practice.
基金This work was supported by the Chinese 973 Program (Grant Nos. G1999011606 & G1999011604).
文摘Application and functional study of dwarf and semi-dwarf genes are of great importance to both crop breeding and molecular biology. A new semi-dwarf gene, sd-t(t), non-allelic to sd-1,had been identified in an indica rice variety, Aitaiyin 2. In this study the gene was genetically mapped by using an F2 population, which consisted of 474 individuals developed from a cross between Aitaiyin 2 and B30. The sd-t(t) gene was located between the RFLP markers R514 and R1408B with a distance of 1.1 cM to R514, and 4.5 cM to R1408B on chromosome 4. A physical contig covering the sd-t(t) mapping region was further constructed by screening a BAC library with R514 and R1408B as probes, and the physical distance between R514 and R1408B was estimated at approximately 147 kb. This result will facilitate map-based cloning of the sd-t(t) gene.
基金supported by the National Natural Science Foundation of China(31991214,91935304,32072055,and 91935302)。
文摘Bread wheat(Triticum aestivum L.,AABBDD,2 n=6 x=42),which accounts for most of the cultivated wheat crop worldwide,is a typical allohexaploid with a genome derived from three diploid wild ancestors.Bread wheat arose and evolved via two sequential allopolyploidization events and was further polished through multiple steps of domestication.Today,cultivated allohexaploid bread wheat has numerous advantageous traits,including adaptive plasticity,favorable yield traits,and extended end-use quality,which have enabled its cultivation well beyond the ranges of its tetraploid and diploid progenitors to become a global staple food crop.In the past decade,rapid advances in wheat genomic research have considerably accelerated our understanding of the bases for the shaping of complex agronomic traits in this polyploid crop.Here,we summarize recent advances in characterizing major genetic factors underlying the origin,evolution,and improvement of polyploid wheats.We end with a brief discussion of the future prospects for the design of gene cloning strategies and modern wheat breeding.
基金supported by China National Transgenic Plant Special Fund(2016ZX08012-002,2016ZX08009-003-007 and 2017ZX08013001-001)to DZ,JS,and ZY,respectivelySMC Morningstar Young Scholarship of Shanghai Jiao Tong University to ZY+2 种基金the Australian Research Council(DP19001941,FT160100218)an IRRTF grant from UoM to SPthe Programme of Introducing Talents of Discipline to Universities(111 Project,B14016)to DZ。
文摘Although Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/CRISPR-associated 9(Cas9)system has been widely used for basic research in model plants,its application for applied breeding in crops has faced strong regulatory obstacles,due mainly to a poor understanding of the authentic output of this system,particularly in higher generations.In this study,different from any previous studies,we investigated in detail the molecular characteristics and production performance of CRISPR/Cas9-generated SD1(semi-dwarf 1)mutants from T2 to T4 generations,of which the selection of T1 and T2 was done only by visual phenotyping for semidwarf plants.Our data revealed not only on-and off-target mutations with small or lager indels but also exogenous elements in T2 plants.All indel mutants passed stably to T3 or T4 without additional modifications independent on the presence of Cas9,while some lines displayed unexpected hereditary patterns of Cas9 or some exogenous elements.In addition,effects of various SD1 alleles on rice height and yield differed depending on genetic backgrounds.Taken together,our data showed that the CRISPR/Cas9 system is effective in producing homozygous mutants for functional analysis,but it may be not as precise as expected in rice,and that early and accurate molecular characterization and screening must be carried out for generations before transitioning of the CRISPR/Cas9 system from laboratory to field.