Clubroot disease,a major plant root disease caused by Plasmodiophora brassicae,has become one of the most destructive diseases among cultivated cruciferous vegetables.However,clubroot-resistant Brassica oleracea mater...Clubroot disease,a major plant root disease caused by Plasmodiophora brassicae,has become one of the most destructive diseases among cultivated cruciferous vegetables.However,clubroot-resistant Brassica oleracea materials are rare.A few clubroot-resistant cabbage varieties are available on the market,but all are Ogura cytoplasmic male sterile(CMS)types.Therefore,in this study,to reutilize the clubroot-resistant Ogura CMS germplasm of cabbage,a new fertility-restored Ogura CMS material,16Q2-11,was used as a bridge to transfer the clubroot resistance(CR)gene from the Ogura CMS cytoplasm to the normal cytoplasm by a two-step method(a fertility restoration and cytoplasm replacement method).In the first cross for fertility restoration of Ogura CMS clubroot-resistant cabbage(FRCRC),16Q2-11 was used as a restorer to cross with Ogura CMS materials containing the CR gene CRb2.Eleven Rfo-positive progenies were generated,of which four contained CRb2:F8-514,F8-620,F8-732 and F8-839.After inoculation with race 4 of P.brassicae,these four CRb2-positive individuals showed resistance.Furthermore,F8-514 and F8-839 were then used as male parents in the second cross of FRCRC to cross with cabbage inbred lines,resulting in the successful introgression of the CRb2 gene into the inbred lines.All offspring produced from this step of cross,which had a normal cytoplasm,showed a high resistance to race 4 of P.brassicae and could be utilized for the breeding of clubrootresistant cabbage varieties in the future.This is the first time that the Ogura CMS restorer has been used to restore the fertility of Ogura CMS clubroot-resistant cabbages,which could improve germplasm diversity in cabbage and provide a reference method for using CMS germplasm in Brassica crops.展开更多
To better understand the genetic diversity and population structure of broccoli cultivars planted in China,a total of 161 representative broccoli cultivars in the past 25 years were collected and analysed based on sin...To better understand the genetic diversity and population structure of broccoli cultivars planted in China,a total of 161 representative broccoli cultivars in the past 25 years were collected and analysed based on single nucleotide polymorphism(SNP)markers.Ten pairs of primers with good polymorphism and high resolution were screened from 315 pairs of SNP primers by 3 broccoli accessions(inbred lines)with different phenotypes and maturity.The 10 pairs of SNP primers were selected,producing 78 alleles.The diversity analysis indicated that the polymorphism information content(PIC)of SNP primer ranged from 0.64 to 0.90.The observed number of alleles(Na)was 2.00,the effective number of alleles(Ne)was 1.11–2.00,the Nei’s gene diversity(H)was 0.10–0.50,and Shannon information index(I)was 0.20–0.70 using PopGene32 software.The clustering results showed that the 161 broccoli cultivars could be divided into 4 major subgroups(A,B,C and D),foreign cultivars were all assigned to subgroup A,and domestic cultivars were assigned to 3 subgroups of B,C,and D.This study indicated that some domestic cultivars and foreign cultivars were similar in genetic background,but most domestic cultivars were still different from the Japanese cultivars.When K=2,the population structure result presented that 161 broccoli cultivars could be divided into 1 simple group(2 groups)and 1 mixed group.When Q≥0.6,143(88.82%)broccoli cultivars belonged to the simple groups.In simple groups 68(42.24%)broccoli cultivars of group 1 were derived from Japan,the United States,Switzerland,the Netherlands,China-Taiwan,and China-Mainland;75(46.58%)broccoli cultivars belonged to group 2;when Q<0.6,18(11.18%)broccoli cultivars belonged to the mixed groups.This study is helpful to understand the diversity and resolution of broccoli cultivars from worldwide,which is beneficial to plant breeding and materials innovation.And meanwhile,this result is also used for construction of broccoli fingerprint serving for cultivar identification.展开更多
Brassica oleracea comprises several important vegetable and ornamental crops,including curly kale,ornamental kale,cabbage,broccoli,and others.The accumulation of anthocyanins,important secondary metabolites valuable t...Brassica oleracea comprises several important vegetable and ornamental crops,including curly kale,ornamental kale,cabbage,broccoli,and others.The accumulation of anthocyanins,important secondary metabolites valuable to human health,in these plants varies widely and is responsible for their pink to dark purple colors.Some curly kale varieties lack anthocyanins,making these plants completely green.The genetic basis of this trait is still unknown.We crossed the curly kale inbred line BK2019(without anthocyanins)with the cabbage inbred line YL1(with anthocyanins)and the Chinese kale inbred line TO1000(with anthocyanins)to generate segregating populations.The no-anthocyanin trait was genetically controlled by a recessive gene,bona1.We generated a linkage map and mapped bona1 to a 256-kb interval on C09.We identified one candidate gene,Bo9g058630,in the target genomic region;this gene is homologous to AT5G42800,which encodes a dihydroflavonol-4-reductase-like(DFR-like)protein in Arabidopsis.In BK2019,a 1-bp insertion was observed in the second exon of Bo9g058630 and directly produced a stop codon.To verify the candidate gene function,CRISPR/Cas9 gene editing technology was applied to knock out Bo9g058630.We generated three bona1 mutants,two of which were completely green with no anthocyanins,confirming that Bo9g058630 corresponds to BoNA1.Different insertion/deletion mutations in BoNA1 exons were found in all six of the other no-anthocyanin kale varieties examined,supporting that independent disruption of BoNA1 resulted in no-anthocyanin varieties of B.oleracea.This study improves the understanding of the regulation mechanism of anthocyanin accumulation in B.oleracea subspecies.展开更多
基金supported by the Major State Research Development Program(2016YFD0101702)the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(CAASASTIP-IVFCAAS)the earmarked fund for the Modern Agro-Industry Technology Research System,China(nycytx-35-gw01).
文摘Clubroot disease,a major plant root disease caused by Plasmodiophora brassicae,has become one of the most destructive diseases among cultivated cruciferous vegetables.However,clubroot-resistant Brassica oleracea materials are rare.A few clubroot-resistant cabbage varieties are available on the market,but all are Ogura cytoplasmic male sterile(CMS)types.Therefore,in this study,to reutilize the clubroot-resistant Ogura CMS germplasm of cabbage,a new fertility-restored Ogura CMS material,16Q2-11,was used as a bridge to transfer the clubroot resistance(CR)gene from the Ogura CMS cytoplasm to the normal cytoplasm by a two-step method(a fertility restoration and cytoplasm replacement method).In the first cross for fertility restoration of Ogura CMS clubroot-resistant cabbage(FRCRC),16Q2-11 was used as a restorer to cross with Ogura CMS materials containing the CR gene CRb2.Eleven Rfo-positive progenies were generated,of which four contained CRb2:F8-514,F8-620,F8-732 and F8-839.After inoculation with race 4 of P.brassicae,these four CRb2-positive individuals showed resistance.Furthermore,F8-514 and F8-839 were then used as male parents in the second cross of FRCRC to cross with cabbage inbred lines,resulting in the successful introgression of the CRb2 gene into the inbred lines.All offspring produced from this step of cross,which had a normal cytoplasm,showed a high resistance to race 4 of P.brassicae and could be utilized for the breeding of clubrootresistant cabbage varieties in the future.This is the first time that the Ogura CMS restorer has been used to restore the fertility of Ogura CMS clubroot-resistant cabbages,which could improve germplasm diversity in cabbage and provide a reference method for using CMS germplasm in Brassica crops.
基金funded by the National Key Research and Development Plan(Grant No.2017YFD0101805)the National Science and Technology Foundation(Grant No.31501761)+2 种基金the National Modern Agricultural Industry Technology System Construction Special Fund Project(Grant No.CARS-23-A8)the Chinese Academy of Agricultural Sciences Science and Technology Innovation Project(Grant No.CAAS-ASTIP-IVF-CAAS)the State Key Laboratory of Vegetable Germplasm Innovation.
文摘To better understand the genetic diversity and population structure of broccoli cultivars planted in China,a total of 161 representative broccoli cultivars in the past 25 years were collected and analysed based on single nucleotide polymorphism(SNP)markers.Ten pairs of primers with good polymorphism and high resolution were screened from 315 pairs of SNP primers by 3 broccoli accessions(inbred lines)with different phenotypes and maturity.The 10 pairs of SNP primers were selected,producing 78 alleles.The diversity analysis indicated that the polymorphism information content(PIC)of SNP primer ranged from 0.64 to 0.90.The observed number of alleles(Na)was 2.00,the effective number of alleles(Ne)was 1.11–2.00,the Nei’s gene diversity(H)was 0.10–0.50,and Shannon information index(I)was 0.20–0.70 using PopGene32 software.The clustering results showed that the 161 broccoli cultivars could be divided into 4 major subgroups(A,B,C and D),foreign cultivars were all assigned to subgroup A,and domestic cultivars were assigned to 3 subgroups of B,C,and D.This study indicated that some domestic cultivars and foreign cultivars were similar in genetic background,but most domestic cultivars were still different from the Japanese cultivars.When K=2,the population structure result presented that 161 broccoli cultivars could be divided into 1 simple group(2 groups)and 1 mixed group.When Q≥0.6,143(88.82%)broccoli cultivars belonged to the simple groups.In simple groups 68(42.24%)broccoli cultivars of group 1 were derived from Japan,the United States,Switzerland,the Netherlands,China-Taiwan,and China-Mainland;75(46.58%)broccoli cultivars belonged to group 2;when Q<0.6,18(11.18%)broccoli cultivars belonged to the mixed groups.This study is helpful to understand the diversity and resolution of broccoli cultivars from worldwide,which is beneficial to plant breeding and materials innovation.And meanwhile,this result is also used for construction of broccoli fingerprint serving for cultivar identification.
基金This work was funded by the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(CAASASTIPIVFCAAS)the China Agriculture Research System of MOF and MARA(CARS-23).
文摘Brassica oleracea comprises several important vegetable and ornamental crops,including curly kale,ornamental kale,cabbage,broccoli,and others.The accumulation of anthocyanins,important secondary metabolites valuable to human health,in these plants varies widely and is responsible for their pink to dark purple colors.Some curly kale varieties lack anthocyanins,making these plants completely green.The genetic basis of this trait is still unknown.We crossed the curly kale inbred line BK2019(without anthocyanins)with the cabbage inbred line YL1(with anthocyanins)and the Chinese kale inbred line TO1000(with anthocyanins)to generate segregating populations.The no-anthocyanin trait was genetically controlled by a recessive gene,bona1.We generated a linkage map and mapped bona1 to a 256-kb interval on C09.We identified one candidate gene,Bo9g058630,in the target genomic region;this gene is homologous to AT5G42800,which encodes a dihydroflavonol-4-reductase-like(DFR-like)protein in Arabidopsis.In BK2019,a 1-bp insertion was observed in the second exon of Bo9g058630 and directly produced a stop codon.To verify the candidate gene function,CRISPR/Cas9 gene editing technology was applied to knock out Bo9g058630.We generated three bona1 mutants,two of which were completely green with no anthocyanins,confirming that Bo9g058630 corresponds to BoNA1.Different insertion/deletion mutations in BoNA1 exons were found in all six of the other no-anthocyanin kale varieties examined,supporting that independent disruption of BoNA1 resulted in no-anthocyanin varieties of B.oleracea.This study improves the understanding of the regulation mechanism of anthocyanin accumulation in B.oleracea subspecies.
