Clubroot caused by Plasmodiophora brassicae is a devastating disease of Cruciferous crops.Developing cultivars with clubroot resistance(CR)is the most effective control measure.For the two major Brassica vegetable spe...Clubroot caused by Plasmodiophora brassicae is a devastating disease of Cruciferous crops.Developing cultivars with clubroot resistance(CR)is the most effective control measure.For the two major Brassica vegetable species B.rapa and B.oleracea,several commercial cultivars with unclear CR pedigrees have been intensively used as CR donors in breeding.However,the continuous occurrence of CR-breaking makes the CR pedigree underlying these cultivars one of the breeders'most urgent concerns.The complex intraspecific diversity of these two major Brassica vegetables has also limited the applicability of CR markers in different breeding programs.Here we first traced the pedigree underlying two kinds of CR that have been widely applied in breeding by linkage and introgression analyses based on public resequencing data.In B.rapa,a major locus CRzi8 underlying the CR of the commercial CR donor‘DegaoCR117’was identified.CRzi8 was further shown to have been introgressed from turnip(B.rapa ssp.rapifera)and that it carried a potential functional allele of Crr1a.The turnip introgression carried CRb^(c),sharing the same coding sequence with the CRb that was also identified from chromosome C07 of B.oleracea CR cultivars with different morphotypes.Within natural populations,variation analysis of linkage intervals of CRzi8,PbBa8.1,CRb,and CRb^(c)yielded easily resolved InDel markers(>20 bp)for these fundamental CR genes.The specificity of these markers was tested in diverse cultivars panels,and each exhibited high reliability in breeding.Our research demonstrates the value of the practice of applying resequencing big data to solve urgent concerns in breeding programs.展开更多
Clubroot disease is a severe threat to Brassica crops globally,particularly in western Canada.Genetic resistance,achieved through pyramiding clubroot resistance(CR)genes with different modes of action,is the most impo...Clubroot disease is a severe threat to Brassica crops globally,particularly in western Canada.Genetic resistance,achieved through pyramiding clubroot resistance(CR)genes with different modes of action,is the most important strategy for managing the disease.However,studies on the CR gene functions are quite limited.In this study,we have conducted investigations into the temporal,structural,and interacting features of a newly cloned CR gene,Rcr1,using CRISPR/Cas9 technology.For temporal functionality,we developed a novel CRISPR/Cas9-based binary vector,pHHIGR-Hsp18.2,to deliver Rcr1 into a susceptible canola line(DH12075)and observed that early expression of Rcr1 is critical for conferring resistance.For structural functionality,several independent mutations in specific domains of Rcr1 resulted in loss-offunction,highlighting their importance for CR phenotype.In the study of the interacting features of Rcr1,a cysteine protease gene and its homologous allele in canola were successfully disrupted via CRISPR/Cas9 as an interacting component with Rcr1 protein,resulting in the conversion from clubroot resistant to susceptible in plants carrying intact Rcr1.These results indicated an indispensable role of these two cysteine proteases in Rcr1-mediated resistance response.This study,the first of its kind,provides valuable insights into the functionality of Rcr1.Further,the new vector p HHIGR-Hsp18.2 demonstrated an inducible feature on the removal of add-on traits,which should be useful for functional genomics and other similar research in brassica crops.展开更多
Chinese cabbage is an economically important Brassica vegetable worldwide, and clubroot, which is caused by the soilborne protist plant pathogen Plasmodiophora brassicae is regarded as a destructive disease to Brassic...Chinese cabbage is an economically important Brassica vegetable worldwide, and clubroot, which is caused by the soilborne protist plant pathogen Plasmodiophora brassicae is regarded as a destructive disease to Brassica crops. Previous studies on the gene transcripts related to Chinese cabbage resistance to clubroot mainly employed RNA-seq technology,although it cannot provide accurate transcript assembly and structural information. In this study, PacBio RS II SMRT sequencing was used to generate full-length transcriptomes of mixed roots at 0, 2, 5, 8, 13, and 22 days after P. brassicae infection in the clubroot-resistant line DH40R. Overall, 39 376 high-quality isoforms and 26 270 open reading frames(ORFs) were identified from the SMRT sequencing data. Additionally, 426 annotated long noncoding RNAs(lncRNAs),56 transcription factor(TF) families, 1 883 genes with poly(A) sites and 1 691 alternative splicing(AS) events were identified. Furthermore, 1 201 of the genes had at least one AS event in DH40R. A comparison with RNA-seq data revealed six differentially expressed AS genes(one for disease resistance and five for defensive response) that are potentially involved in P. brassicae resistance. The results of this study provide valuable resources for basic research on clubroot resistance in Chinese cabbage.展开更多
Clubroot and herbicide resistance,high oleic acid(OA)content,and early maturity are targets of rapeseed(Brassica napus L.)breeding.The objective of this study was to develop new male-fertility restorer lines by pyrami...Clubroot and herbicide resistance,high oleic acid(OA)content,and early maturity are targets of rapeseed(Brassica napus L.)breeding.The objective of this study was to develop new male-fertility restorer lines by pyramiding favorable genes to improve these traits simultaneously.Seven elite alleles for the four traits were introduced into the restorer line 621R by speed breeding with marker-assisted and phenotypic selection.Six introgression lines(ILs)were developed with four-to seven-gene combinations and crossed with two elite parents to develop hybrids.All ILs and their corresponding hybrids displayed high resistance to both clubroot pathotype 4 and sulfonylurea herbicides.Three ILs and their hybrids showed large increases in OA contents and four showed earlier maturity.These new ILs may be useful in rapeseed hybrid breeding for the target traits.展开更多
Clubroot disease, caused by Plasmodiophora brassicae, is one of the most destructive soil-borne diseases in cruciferous crops worldwide. New strategies are urgently needed to control this disease, as no effective dise...Clubroot disease, caused by Plasmodiophora brassicae, is one of the most destructive soil-borne diseases in cruciferous crops worldwide. New strategies are urgently needed to control this disease, as no effective disease-resistant varieties or chemical control agents exist. Previously, we found that the incidence rate and disease index of clubroot in oilseed rape decreased by 50 and 40%, respectively, when oilseed rape was planted after soybean. In order to understand how different rotation patterns affect the occurrence of clubroot in oilseed rape, high-throughput sequencing was used to analyze the rhizosphere microbial community of oilseed rape planted after leguminous (soybean, clover), gramineous (rice, maize) and cruciferous (oilseed rape, Chinese cabbage) crops. Results showed that planting soybeans before oilseed rape significantly increased the population density of microbes that could inhibit P. brassicae (e.g., Sphingomonas, Bacillus, Streptomyces and Trichoderma). Conversely, consecutive cultivation of cruciferous crops significantly accumulated plant pathogens, including P. brassicae, Olpidium and Colletotrichum (P<0.05). These results will help to develop the most effective rotation pattern for reducing clubroot damage.展开更多
In this study, using Japanese radish cytoplasmic sterile line "Half Green Cabbage" (HGC) and clubfoot-resistant South Korean cabbage variety " CR Cabbage King" (CCK) as experimental materials, serf-incompatibl...In this study, using Japanese radish cytoplasmic sterile line "Half Green Cabbage" (HGC) and clubfoot-resistant South Korean cabbage variety " CR Cabbage King" (CCK) as experimental materials, serf-incompatible DH lines were obtained by mierosporo culture. Through five gnerations of backeross, CCR11239, a clubroot-resistant radish cytoplasmic sterile line, was obtained. Through six generations of serf-crossing, CCR11240, the maintainer line of CCR11239, was bred. After the cross of CCK and Luchunbai 1 (83 - 1 ) and five generations of self-crossing, serf-compatible line CCRl1241 was obtained. Through crossing CCR11240 with CCRl1241, the new cabbage variety CCRl1242 was obtained. After variety comparison test, provincial regional test and production demonstration test, the new variety was registered in 2012. The average yield of CCR11242 reached 60 975 kg/hm2, which was improved by 142% compared with Luchunbai 1 (83 - 1 ). The disease index of CCR11242 was 5.63, which was 88.13 lower than the control, indicating high resistance (HR) to clubfoot.展开更多
The transcriptome-wide gene expression was compared between susceptible and resistant Chinese cabbage cultivars to identify genes that contributed to clubroot resistance. A higher number of differentially expressed ge...The transcriptome-wide gene expression was compared between susceptible and resistant Chinese cabbage cultivars to identify genes that contributed to clubroot resistance. A higher number of differentially expressed genes were detected in susceptible cultivars than in resistant cultivars. Fifty-six genes involved in cell wall modification, hormone signaling, root marphogenesis, nematodes response and cell proliferation were uniquely expressed in the resistant cultivars. Among them, 27 genes were involved in cell wall modification and hormone signaling, indicating that genes in these two types might play a vital role in the defense response to pathogen infection.展开更多
[Objective] The paper was to analyze the inheritance of clubroot resistance of miniature Chinese baby cabbage,to shorten identification time and to improve breeding efficacy of new disease-resistant varieties. [Method...[Objective] The paper was to analyze the inheritance of clubroot resistance of miniature Chinese baby cabbage,to shorten identification time and to improve breeding efficacy of new disease-resistant varieties. [Method] Taking clubroot-resistant Chinese cabbage CCR001,disease-susceptible baby cabbage CM002,and their F_1,F_2 and BC_1 offspring as the research objects,the inheritance of clubroot resistance of baby cabbage was studied. [Result]The clubroot-resistance of baby cabbage was controlled by a single dominant gene,which conformed to Mendel's Laws of inheritance. The molecular markers-assisted selection combing with bacterial soil inoculation confirmed that the disease-resistance indeed passed on from the parents,and was inherited in F_1 and F_2. [Conclusion]It is feasible to breed clubroot-resistant baby cabbage by using molecular markers-assisted selection.展开更多
Clubroot is a prevailing soil-borne disease affecting rapeseed production worldwide.However,few clubroot resistant rapeseed accessions were available for breeding.Identification and introgression of new clubroot resis...Clubroot is a prevailing soil-borne disease affecting rapeseed production worldwide.However,few clubroot resistant rapeseed accessions were available for breeding.Identification and introgression of new clubroot resistant genes from closely related species by distant hybridization is an effective strategy.In the present study,9 radish(Raphanus sativus L.,2n=18,RR)lines resistant to Plasmodiophora brassicae pathotype 4 were used as donors to transfer clubroot resistance into a susceptible rapeseed(Brassica napus L.,2n=38,AACC)line by distant hybridization combined with embryo rescue.Nine intergeneric crosses were made but only 1(411×93039)produced F1 plants both from embryo rescue and natural seed-setting.Authenticity of triploid F1 hybrids(2n=28,ACR)were verified by flower color,cytological observation and molecular marker analysis,and 2 genuine F1 hybrids were identified.After chromosome doubling,these synthetic allohexaploid plants(2n=56,AACCRR)became partially fertile(pollen viability rate=35%)and were backcrossed with rapeseed parent to generate a BC1 population(2n=47,AACCR).Totally 178 BC1 plants were obtained,of which the majority(96.1%)were resistant to clubroot.These backcrossing progenies could be used for the breeding of new rapeseed varieties resistant to clubroot.展开更多
Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection.Breeding of clubroot resistant cultivars bas...Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection.Breeding of clubroot resistant cultivars based on mapping and cloning of resistant genes is commonly regarded as the most cost-effective and efficient way to fight against this disease.The traditional way of R gene functional validation requires stable transformation that is both time-and labor-consuming.In this study,a rapid and efficient hairy-root transgenic protocol mediated by Agrobacterium rhizogenes was developed.The transformation positive rate was over 80%in Brassica napus showed by GUS reporter gene and this transformation only took 1/6 of the time compared with stable transformation.The system was applicable to different B.napus varieties and other cruciferous crops including Brassica rapa and Brassica oleracea.In particular,two known CR genes,CRA3.7.1 and CRA8.2.4 were used respectively,as example to show that the system works well for CR gene study combined with subsequent P.brassicae infection in B.napus.Most importantly,it works both in over-expression that led to disease resistance,as well as in RNAi which led to disease susceptible phenotype.Therefore,this system can be used in batch-wise identification of CR genes,and also offered the possibility of manipulating key genes within the P.brassicae genome that could improve our knowledge on host-pathogen interaction.展开更多
通过远缘杂交及回交将白菜的抗病基因导入到芥菜中,创制获得优质的抗根肿病长柄芥(Brassica juncea var.longepetiolata Yang et Chen)育种材料。采用人工接种鉴定和分子标记鉴定相结合法,对10份抗根肿病白菜材料进行抗性分析,筛选出1...通过远缘杂交及回交将白菜的抗病基因导入到芥菜中,创制获得优质的抗根肿病长柄芥(Brassica juncea var.longepetiolata Yang et Chen)育种材料。采用人工接种鉴定和分子标记鉴定相结合法,对10份抗根肿病白菜材料进行抗性分析,筛选出1份可作为长柄芥抗根肿病育种材料创制的抗性资源CCR21002。利用CCR21002分别作为父本、母本提供抗根肿病基因,与长柄芥进行远缘杂交,结合胚挽救技术获得正反杂交种F1,选取植株形态偏向芥菜的F1与长柄芥回交获得BC_(1),对BC_(1)植株进行形态学、细胞学分析及抗病性鉴定。结果表明,回交一代比杂种一代更难获得,胚挽救成活的13株BC_(1)中,仅筛选出1株基因型为AABB且抗根肿病的植株,经分子标记鉴定所含抗病基因为CRb,在植株形态上保留有长柄芥的大部分性状。证明经过与抗根肿病白菜远缘杂交及回交,获得对4号生理小种表现抗性的长柄芥新种质1份,可用于继续回交转育。展开更多
基金supported by the China Agriculture Research System(Grant No.CARS-23-A13)Hubei Agrotechnical Major Project(Grant No.2021-620-000-001-01)+1 种基金Wuhan Major Project of Key Technologies in Biological Breeding and New Variety Cultivation(Grant No.2022021302024852)HZAU-AGIS Cooperation Fund(Grant No.SZYJY2023022).
文摘Clubroot caused by Plasmodiophora brassicae is a devastating disease of Cruciferous crops.Developing cultivars with clubroot resistance(CR)is the most effective control measure.For the two major Brassica vegetable species B.rapa and B.oleracea,several commercial cultivars with unclear CR pedigrees have been intensively used as CR donors in breeding.However,the continuous occurrence of CR-breaking makes the CR pedigree underlying these cultivars one of the breeders'most urgent concerns.The complex intraspecific diversity of these two major Brassica vegetables has also limited the applicability of CR markers in different breeding programs.Here we first traced the pedigree underlying two kinds of CR that have been widely applied in breeding by linkage and introgression analyses based on public resequencing data.In B.rapa,a major locus CRzi8 underlying the CR of the commercial CR donor‘DegaoCR117’was identified.CRzi8 was further shown to have been introgressed from turnip(B.rapa ssp.rapifera)and that it carried a potential functional allele of Crr1a.The turnip introgression carried CRb^(c),sharing the same coding sequence with the CRb that was also identified from chromosome C07 of B.oleracea CR cultivars with different morphotypes.Within natural populations,variation analysis of linkage intervals of CRzi8,PbBa8.1,CRb,and CRb^(c)yielded easily resolved InDel markers(>20 bp)for these fundamental CR genes.The specificity of these markers was tested in diverse cultivars panels,and each exhibited high reliability in breeding.Our research demonstrates the value of the practice of applying resequencing big data to solve urgent concerns in breeding programs.
基金supported by the Genomics Initiative of Agriculture and Agri-Food Canada。
文摘Clubroot disease is a severe threat to Brassica crops globally,particularly in western Canada.Genetic resistance,achieved through pyramiding clubroot resistance(CR)genes with different modes of action,is the most important strategy for managing the disease.However,studies on the CR gene functions are quite limited.In this study,we have conducted investigations into the temporal,structural,and interacting features of a newly cloned CR gene,Rcr1,using CRISPR/Cas9 technology.For temporal functionality,we developed a novel CRISPR/Cas9-based binary vector,pHHIGR-Hsp18.2,to deliver Rcr1 into a susceptible canola line(DH12075)and observed that early expression of Rcr1 is critical for conferring resistance.For structural functionality,several independent mutations in specific domains of Rcr1 resulted in loss-offunction,highlighting their importance for CR phenotype.In the study of the interacting features of Rcr1,a cysteine protease gene and its homologous allele in canola were successfully disrupted via CRISPR/Cas9 as an interacting component with Rcr1 protein,resulting in the conversion from clubroot resistant to susceptible in plants carrying intact Rcr1.These results indicated an indispensable role of these two cysteine proteases in Rcr1-mediated resistance response.This study,the first of its kind,provides valuable insights into the functionality of Rcr1.Further,the new vector p HHIGR-Hsp18.2 demonstrated an inducible feature on the removal of add-on traits,which should be useful for functional genomics and other similar research in brassica crops.
基金supported by the National Natural Science Foundation of China (31872945 and 31801874)the earmarked fund for China Agricultural Research System (CARS-23-G15)+1 种基金the Funds for Distinguished Young Scientists from Henan Academy of Agricultural Sciences, China (2021JQ03)the Innovation Team of Henan Academy of Agricultural Sciences, China (2021TD06)。
文摘Chinese cabbage is an economically important Brassica vegetable worldwide, and clubroot, which is caused by the soilborne protist plant pathogen Plasmodiophora brassicae is regarded as a destructive disease to Brassica crops. Previous studies on the gene transcripts related to Chinese cabbage resistance to clubroot mainly employed RNA-seq technology,although it cannot provide accurate transcript assembly and structural information. In this study, PacBio RS II SMRT sequencing was used to generate full-length transcriptomes of mixed roots at 0, 2, 5, 8, 13, and 22 days after P. brassicae infection in the clubroot-resistant line DH40R. Overall, 39 376 high-quality isoforms and 26 270 open reading frames(ORFs) were identified from the SMRT sequencing data. Additionally, 426 annotated long noncoding RNAs(lncRNAs),56 transcription factor(TF) families, 1 883 genes with poly(A) sites and 1 691 alternative splicing(AS) events were identified. Furthermore, 1 201 of the genes had at least one AS event in DH40R. A comparison with RNA-seq data revealed six differentially expressed AS genes(one for disease resistance and five for defensive response) that are potentially involved in P. brassicae resistance. The results of this study provide valuable resources for basic research on clubroot resistance in Chinese cabbage.
基金supported by the China Agriculture Research System of MOF and MARA(CARS-12)the Open Fund of the National Key Laboratory of Crop Genetic Improvement(ZK201909)。
文摘Clubroot and herbicide resistance,high oleic acid(OA)content,and early maturity are targets of rapeseed(Brassica napus L.)breeding.The objective of this study was to develop new male-fertility restorer lines by pyramiding favorable genes to improve these traits simultaneously.Seven elite alleles for the four traits were introduced into the restorer line 621R by speed breeding with marker-assisted and phenotypic selection.Six introgression lines(ILs)were developed with four-to seven-gene combinations and crossed with two elite parents to develop hybrids.All ILs and their corresponding hybrids displayed high resistance to both clubroot pathotype 4 and sulfonylurea herbicides.Three ILs and their hybrids showed large increases in OA contents and four showed earlier maturity.These new ILs may be useful in rapeseed hybrid breeding for the target traits.
基金This work was supported by the National Key Research and Development Program of China(2017YFD0200600)the Financial Innovation Capacity Enhancement Project in Sichuan Province,China(2019QNJJ-011)the National Modern Agricultural Industry technology System of Sichuan Rape Innovation Team,China(2019-2023).
文摘Clubroot disease, caused by Plasmodiophora brassicae, is one of the most destructive soil-borne diseases in cruciferous crops worldwide. New strategies are urgently needed to control this disease, as no effective disease-resistant varieties or chemical control agents exist. Previously, we found that the incidence rate and disease index of clubroot in oilseed rape decreased by 50 and 40%, respectively, when oilseed rape was planted after soybean. In order to understand how different rotation patterns affect the occurrence of clubroot in oilseed rape, high-throughput sequencing was used to analyze the rhizosphere microbial community of oilseed rape planted after leguminous (soybean, clover), gramineous (rice, maize) and cruciferous (oilseed rape, Chinese cabbage) crops. Results showed that planting soybeans before oilseed rape significantly increased the population density of microbes that could inhibit P. brassicae (e.g., Sphingomonas, Bacillus, Streptomyces and Trichoderma). Conversely, consecutive cultivation of cruciferous crops significantly accumulated plant pathogens, including P. brassicae, Olpidium and Colletotrichum (P<0.05). These results will help to develop the most effective rotation pattern for reducing clubroot damage.
基金Supported by Major Project of New Product Development of Yunnan Provincial Science and Technology Department(2015BB007,2012BB017)International Cooperation Project of Yunnan Provincial Science and Technology Department(2014IA016)+2 种基金Project of Science and Technology to Strengthen the County and Enrich the People of Yunnan Provincial Science and Technology Department(2014EB033)National Bulk Vegetable Industry Technology System of China(CARS-25-G-45)New Vegetable Variety Cooperation Project of Yunnan Provincial Department of Agriculture[YCN(2012)No.58]
文摘In this study, using Japanese radish cytoplasmic sterile line "Half Green Cabbage" (HGC) and clubfoot-resistant South Korean cabbage variety " CR Cabbage King" (CCK) as experimental materials, serf-incompatible DH lines were obtained by mierosporo culture. Through five gnerations of backeross, CCR11239, a clubroot-resistant radish cytoplasmic sterile line, was obtained. Through six generations of serf-crossing, CCR11240, the maintainer line of CCR11239, was bred. After the cross of CCK and Luchunbai 1 (83 - 1 ) and five generations of self-crossing, serf-compatible line CCRl1241 was obtained. Through crossing CCR11240 with CCRl1241, the new cabbage variety CCRl1242 was obtained. After variety comparison test, provincial regional test and production demonstration test, the new variety was registered in 2012. The average yield of CCR11242 reached 60 975 kg/hm2, which was improved by 142% compared with Luchunbai 1 (83 - 1 ). The disease index of CCR11242 was 5.63, which was 88.13 lower than the control, indicating high resistance (HR) to clubfoot.
基金Supported by the National Key Research and Development Project(2017YFD0101803)
文摘The transcriptome-wide gene expression was compared between susceptible and resistant Chinese cabbage cultivars to identify genes that contributed to clubroot resistance. A higher number of differentially expressed genes were detected in susceptible cultivars than in resistant cultivars. Fifty-six genes involved in cell wall modification, hormone signaling, root marphogenesis, nematodes response and cell proliferation were uniquely expressed in the resistant cultivars. Among them, 27 genes were involved in cell wall modification and hormone signaling, indicating that genes in these two types might play a vital role in the defense response to pathogen infection.
基金Supported by Key New Product Plan of Yunnan Provincial Science and Technology Department(2015BB007)International Cooperation Project of Yunnan Provincial Science and Technology Department(2014IA016)+1 种基金Science and Technology Plan for Enriching People and Strengthening County of Yunnan Provincial Science and Technology Department(2014EB033)National Bulk Vegetable Industry Technology System(CARS-23)
文摘[Objective] The paper was to analyze the inheritance of clubroot resistance of miniature Chinese baby cabbage,to shorten identification time and to improve breeding efficacy of new disease-resistant varieties. [Method] Taking clubroot-resistant Chinese cabbage CCR001,disease-susceptible baby cabbage CM002,and their F_1,F_2 and BC_1 offspring as the research objects,the inheritance of clubroot resistance of baby cabbage was studied. [Result]The clubroot-resistance of baby cabbage was controlled by a single dominant gene,which conformed to Mendel's Laws of inheritance. The molecular markers-assisted selection combing with bacterial soil inoculation confirmed that the disease-resistance indeed passed on from the parents,and was inherited in F_1 and F_2. [Conclusion]It is feasible to breed clubroot-resistant baby cabbage by using molecular markers-assisted selection.
基金Financial support from the National Key Research and the Development Program of China (2018YFD0200902, 2016YFD0100202)
文摘Clubroot is a prevailing soil-borne disease affecting rapeseed production worldwide.However,few clubroot resistant rapeseed accessions were available for breeding.Identification and introgression of new clubroot resistant genes from closely related species by distant hybridization is an effective strategy.In the present study,9 radish(Raphanus sativus L.,2n=18,RR)lines resistant to Plasmodiophora brassicae pathotype 4 were used as donors to transfer clubroot resistance into a susceptible rapeseed(Brassica napus L.,2n=38,AACC)line by distant hybridization combined with embryo rescue.Nine intergeneric crosses were made but only 1(411×93039)produced F1 plants both from embryo rescue and natural seed-setting.Authenticity of triploid F1 hybrids(2n=28,ACR)were verified by flower color,cytological observation and molecular marker analysis,and 2 genuine F1 hybrids were identified.After chromosome doubling,these synthetic allohexaploid plants(2n=56,AACCRR)became partially fertile(pollen viability rate=35%)and were backcrossed with rapeseed parent to generate a BC1 population(2n=47,AACCR).Totally 178 BC1 plants were obtained,of which the majority(96.1%)were resistant to clubroot.These backcrossing progenies could be used for the breeding of new rapeseed varieties resistant to clubroot.
基金supported by grants from the Wuhan Science and Technology Major Project on Key techniques of biological breeding and Breeding of new varieties(Grant No.2022021302024851)the special project for sustainable development agenda of innovation demonstration zone(Grant No.202204AC100001-A04)the National Key R&D Program of China(Grant No.2022YFD1200400)。
文摘Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection.Breeding of clubroot resistant cultivars based on mapping and cloning of resistant genes is commonly regarded as the most cost-effective and efficient way to fight against this disease.The traditional way of R gene functional validation requires stable transformation that is both time-and labor-consuming.In this study,a rapid and efficient hairy-root transgenic protocol mediated by Agrobacterium rhizogenes was developed.The transformation positive rate was over 80%in Brassica napus showed by GUS reporter gene and this transformation only took 1/6 of the time compared with stable transformation.The system was applicable to different B.napus varieties and other cruciferous crops including Brassica rapa and Brassica oleracea.In particular,two known CR genes,CRA3.7.1 and CRA8.2.4 were used respectively,as example to show that the system works well for CR gene study combined with subsequent P.brassicae infection in B.napus.Most importantly,it works both in over-expression that led to disease resistance,as well as in RNAi which led to disease susceptible phenotype.Therefore,this system can be used in batch-wise identification of CR genes,and also offered the possibility of manipulating key genes within the P.brassicae genome that could improve our knowledge on host-pathogen interaction.
文摘通过远缘杂交及回交将白菜的抗病基因导入到芥菜中,创制获得优质的抗根肿病长柄芥(Brassica juncea var.longepetiolata Yang et Chen)育种材料。采用人工接种鉴定和分子标记鉴定相结合法,对10份抗根肿病白菜材料进行抗性分析,筛选出1份可作为长柄芥抗根肿病育种材料创制的抗性资源CCR21002。利用CCR21002分别作为父本、母本提供抗根肿病基因,与长柄芥进行远缘杂交,结合胚挽救技术获得正反杂交种F1,选取植株形态偏向芥菜的F1与长柄芥回交获得BC_(1),对BC_(1)植株进行形态学、细胞学分析及抗病性鉴定。结果表明,回交一代比杂种一代更难获得,胚挽救成活的13株BC_(1)中,仅筛选出1株基因型为AABB且抗根肿病的植株,经分子标记鉴定所含抗病基因为CRb,在植株形态上保留有长柄芥的大部分性状。证明经过与抗根肿病白菜远缘杂交及回交,获得对4号生理小种表现抗性的长柄芥新种质1份,可用于继续回交转育。