[目的]本文旨在分析播栽温度对水稻野败型细胞质雄性不育性(wild abortive type cytoplasmic male sterility,CMS-WA)的影响,为不育系选育、提纯、繁殖和制种提供理论依据。[方法]设置16.8、21.2、24.5、27.3、30.1℃5个播栽温度,在自...[目的]本文旨在分析播栽温度对水稻野败型细胞质雄性不育性(wild abortive type cytoplasmic male sterility,CMS-WA)的影响,为不育系选育、提纯、繁殖和制种提供理论依据。[方法]设置16.8、21.2、24.5、27.3、30.1℃5个播栽温度,在自然温度梯度压力下,选择3个不同微效恢复基因数量的野败型(wild abortion type,WA)不育系珍汕97A、龙特浦A和京福1A,以嵌合颖花率和黑染花粉率为指标进行对比研究,应用回归模型和主效可加互作可乘模型(additive main effects and multiplicative interaction,AMMI),分析不育系花粉育性的变化动态。[结果]随着播栽期的推迟,不育系播始历期缩短,生育进程加快,有效积温总体呈先升后降趋势,表现感温性。播栽温度在20.1~30.1℃时能提高不育系黑染花粉率和嵌合颖花率,呈正效应。随着播栽温度的上升,3个不育系的黑染花粉率和嵌合颖花率总体上均表现上升趋势,龙特浦A响应极显著,京福1A和珍汕97A表现不显著。珍汕97A花粉育性平均值较低,黑染花粉率和嵌合颖花率分别为1.398%和5.628%,其基因型互作效应主成分轴(interaction principal component axis,ICPA1)值分别为-1.632和-1.512,表明其不育性稳定,花粉育性对播栽温度敏感度低;龙特浦A和京福1A花粉育性平均值较高,二者的黑染花粉率分别为9.836%和7.580%,嵌合颖花率分别为14.788%和11.810%,黑染花粉率基因型IPCA1数值分别为1.114和0.518,嵌合颖花率基因型IPCA1数值分别为0.133和1.379,表明其不育性较不稳定,花粉育性对播栽温度变化相对敏感。[结论]根据温度对不育系育性影响的变化规律,对于温度敏感型的不育系,宜采取早繁早制的措施,保证种子纯度。在不育系选育和遗传提纯上,应在微效恢复基因高效表达的温度条件下进行,可及早剔除不合格单株或株系,以获得不育性较稳定后代。展开更多
The discovery of a wild abortive-type(WA)cytoplasmic male sterile(CMS)line and breeding its restorer line have led to the commercialization of three-line hybrid rice,contributing considerably to global food security.H...The discovery of a wild abortive-type(WA)cytoplasmic male sterile(CMS)line and breeding its restorer line have led to the commercialization of three-line hybrid rice,contributing considerably to global food security.However,the molecular mechanisms underlyingfertility abortion and the restoration of CMs-WA lines remain largely elusive.In this study,we cloned a restorer gene,Rf2o,following a genome-wide association study analysis of the core parent lines of three-line hybrid rice.We found that Rf20 was present in all core parental lines,but different haplotypes and structural variants of its gene resulted in differences in Rf2o expression levels between sterileand restored lines.Rf20 could restorepollen fertility in the CMS-WA line and was found to be responsible for fertility restoration in some CMs lines under high temperatures.In addition,we found that Rf20 encodes a pentatricopeptide repeat protein that competes with WA352 for binding with COX11.This interaction enhances Cox11's function as a scavenger of reactive oxygen species,which in turn restores pollenfertility.Collectively,ourstudysuggestsanewactionmodeforpentatricopeptide repeatproteins inthe fertilityrestorationofCMs lines,providing an essential theoretical basisforbreedingrobustrestorer lines and for overcoming high temperature-induced fertility recovery of some CMS lines.展开更多
The use of cytoplasm male sterility （CMS） is crucial for three-line hybrid seed production. Two types of CMS have been discovered from Dongxiang wild rice, namely the wild abortion type CMS （CMS-WA） and the Dongxi...The use of cytoplasm male sterility （CMS） is crucial for three-line hybrid seed production. Two types of CMS have been discovered from Dongxiang wild rice, namely the wild abortion type CMS （CMS-WA） and the Dongxiang wild type CMS （CMS-DW）. In this study, we show the molecular identification of the two types of CMS in Dongxiang wild rice. WA352, which conferred CMS-WA, was not detected in Dongxiang wild iice, implying Dongxiang wild rice does not carry the CMS- WA source. Further analysis of WA352 in DYIA, a CMS-DW line, by PCR amplification and sequencing, revealed two insertion-deletion polymorphisms occurred in CMS-DW compared to WA352 of CMS-WA. It was reported that WA352 was comprised of an unknown origin sequence and partial sequences of three open reading frames （ORFs）, orf284, orf224 and orf288. The 42-bp insertion was located between the two segments of orf224 and orf288, which created a new chimeric ORF, orf216. This new ORF was also detected in CMS-HL. Based on the 9-bp deletion in orf284, a specific mitochon- drial marker of DW-M1 was developed, which could be used to specifically distinguish the DW-type source. Moreover, semi-quantitative RT-PCR analysis preliminarily suggested that both orf216 and orf284 could be considered as candidates for CMS-DW. These findings present a preliminary understanding of CMS-DW at the molecular level.展开更多
RNA editing changes the nucleotides at the transcript level of mitochondrial genes which results in synthesis of functional proteins.This study was designed to find the editing sites which could be implicated in male ...RNA editing changes the nucleotides at the transcript level of mitochondrial genes which results in synthesis of functional proteins.This study was designed to find the editing sites which could be implicated in male fertility restoration and to develop editing based markers for differentiation of cytoplasmic male sterility and maintainer lines from each other.DNA and RNA from young panicles were isolated from three-line system of hybrid rice PRH10,wild abortive(WA)cytoplasm based male sterile(A line Pusa 6A),maintainer(B line Pusa 6B)and restorer(R line PRR78)lines.Pusa 6A and PRR78 having the same WA cytoplasm are allo-nuclear and iso-cytpolasmic lines.The genomic and cDNA amplicons for eight mitochondrial genes(18SrRNA,atp6,atp9,cobII,coxI,coxIII,nadI and rps3)were sequenced and compared.Differences in genomic and cDNA sequences were considered as editing.Two hundred and thirty editing sites having base substitution or insertion/deletion were identified with the highest in 18SrRNA(5.74%)and the lowest in coxI(0.60%).The highest editing sites were observed in fertile maintainer Pusa 6B followed by PRR78 and Pusa 6A,of which random five editing sites in five different rice mitochondrial transcripts namely atp9,cobII,coxIII,rps3 and 18SrRNA were chosen and validated through cleaved amplified polymorphism sequence(CAPS)analysis and found to be partially edited in four genes.The identical editing sites of different mitochondrial genes from maintainer and restorer lines might reflect their possible contribution to fertility restoration of sterile WA cytoplasm.展开更多
文摘[目的]本文旨在分析播栽温度对水稻野败型细胞质雄性不育性(wild abortive type cytoplasmic male sterility,CMS-WA)的影响,为不育系选育、提纯、繁殖和制种提供理论依据。[方法]设置16.8、21.2、24.5、27.3、30.1℃5个播栽温度,在自然温度梯度压力下,选择3个不同微效恢复基因数量的野败型(wild abortion type,WA)不育系珍汕97A、龙特浦A和京福1A,以嵌合颖花率和黑染花粉率为指标进行对比研究,应用回归模型和主效可加互作可乘模型(additive main effects and multiplicative interaction,AMMI),分析不育系花粉育性的变化动态。[结果]随着播栽期的推迟,不育系播始历期缩短,生育进程加快,有效积温总体呈先升后降趋势,表现感温性。播栽温度在20.1~30.1℃时能提高不育系黑染花粉率和嵌合颖花率,呈正效应。随着播栽温度的上升,3个不育系的黑染花粉率和嵌合颖花率总体上均表现上升趋势,龙特浦A响应极显著,京福1A和珍汕97A表现不显著。珍汕97A花粉育性平均值较低,黑染花粉率和嵌合颖花率分别为1.398%和5.628%,其基因型互作效应主成分轴(interaction principal component axis,ICPA1)值分别为-1.632和-1.512,表明其不育性稳定,花粉育性对播栽温度敏感度低;龙特浦A和京福1A花粉育性平均值较高,二者的黑染花粉率分别为9.836%和7.580%,嵌合颖花率分别为14.788%和11.810%,黑染花粉率基因型IPCA1数值分别为1.114和0.518,嵌合颖花率基因型IPCA1数值分别为0.133和1.379,表明其不育性较不稳定,花粉育性对播栽温度变化相对敏感。[结论]根据温度对不育系育性影响的变化规律,对于温度敏感型的不育系,宜采取早繁早制的措施,保证种子纯度。在不育系选育和遗传提纯上,应在微效恢复基因高效表达的温度条件下进行,可及早剔除不合格单株或株系,以获得不育性较稳定后代。
基金Biological Breeding-Major Projects(2023ZD04076)National Key Research and Development Project(2022YFD1200800)+1 种基金Hunan Natural Science Foundation(2023JJ30442)Special Funds for the Construction of Innovative Provinces in Hunan Province(2021NK1002).
文摘The discovery of a wild abortive-type(WA)cytoplasmic male sterile(CMS)line and breeding its restorer line have led to the commercialization of three-line hybrid rice,contributing considerably to global food security.However,the molecular mechanisms underlyingfertility abortion and the restoration of CMs-WA lines remain largely elusive.In this study,we cloned a restorer gene,Rf2o,following a genome-wide association study analysis of the core parent lines of three-line hybrid rice.We found that Rf20 was present in all core parental lines,but different haplotypes and structural variants of its gene resulted in differences in Rf2o expression levels between sterileand restored lines.Rf20 could restorepollen fertility in the CMS-WA line and was found to be responsible for fertility restoration in some CMs lines under high temperatures.In addition,we found that Rf20 encodes a pentatricopeptide repeat protein that competes with WA352 for binding with COX11.This interaction enhances Cox11's function as a scavenger of reactive oxygen species,which in turn restores pollenfertility.Collectively,ourstudysuggestsanewactionmodeforpentatricopeptide repeatproteins inthe fertilityrestorationofCMs lines,providing an essential theoretical basisforbreedingrobustrestorer lines and for overcoming high temperature-induced fertility recovery of some CMS lines.
基金supported by the National Natural Science Foundation of China (31460343)the Key Program for Science and Technology of Jiangxi Province, China (20152ACF60015)
文摘The use of cytoplasm male sterility （CMS） is crucial for three-line hybrid seed production. Two types of CMS have been discovered from Dongxiang wild rice, namely the wild abortion type CMS （CMS-WA） and the Dongxiang wild type CMS （CMS-DW）. In this study, we show the molecular identification of the two types of CMS in Dongxiang wild rice. WA352, which conferred CMS-WA, was not detected in Dongxiang wild iice, implying Dongxiang wild rice does not carry the CMS- WA source. Further analysis of WA352 in DYIA, a CMS-DW line, by PCR amplification and sequencing, revealed two insertion-deletion polymorphisms occurred in CMS-DW compared to WA352 of CMS-WA. It was reported that WA352 was comprised of an unknown origin sequence and partial sequences of three open reading frames （ORFs）, orf284, orf224 and orf288. The 42-bp insertion was located between the two segments of orf224 and orf288, which created a new chimeric ORF, orf216. This new ORF was also detected in CMS-HL. Based on the 9-bp deletion in orf284, a specific mitochon- drial marker of DW-M1 was developed, which could be used to specifically distinguish the DW-type source. Moreover, semi-quantitative RT-PCR analysis preliminarily suggested that both orf216 and orf284 could be considered as candidates for CMS-DW. These findings present a preliminary understanding of CMS-DW at the molecular level.
文摘RNA editing changes the nucleotides at the transcript level of mitochondrial genes which results in synthesis of functional proteins.This study was designed to find the editing sites which could be implicated in male fertility restoration and to develop editing based markers for differentiation of cytoplasmic male sterility and maintainer lines from each other.DNA and RNA from young panicles were isolated from three-line system of hybrid rice PRH10,wild abortive(WA)cytoplasm based male sterile(A line Pusa 6A),maintainer(B line Pusa 6B)and restorer(R line PRR78)lines.Pusa 6A and PRR78 having the same WA cytoplasm are allo-nuclear and iso-cytpolasmic lines.The genomic and cDNA amplicons for eight mitochondrial genes(18SrRNA,atp6,atp9,cobII,coxI,coxIII,nadI and rps3)were sequenced and compared.Differences in genomic and cDNA sequences were considered as editing.Two hundred and thirty editing sites having base substitution or insertion/deletion were identified with the highest in 18SrRNA(5.74%)and the lowest in coxI(0.60%).The highest editing sites were observed in fertile maintainer Pusa 6B followed by PRR78 and Pusa 6A,of which random five editing sites in five different rice mitochondrial transcripts namely atp9,cobII,coxIII,rps3 and 18SrRNA were chosen and validated through cleaved amplified polymorphism sequence(CAPS)analysis and found to be partially edited in four genes.The identical editing sites of different mitochondrial genes from maintainer and restorer lines might reflect their possible contribution to fertility restoration of sterile WA cytoplasm.
