以分子标记辅助连续回交快速提高花生品种油酸含量及对其后代农艺性状的评价
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摘要
高油酸是花生重要的品质性状,高油酸花生及其制品具有较好的品质稳定性和较高的营养和保健价值。我国高油酸花生的育成品种类型较少,遗传背景不够丰富,育种手段比较单一。针对上述问题,本研究开发了AS-PCR-MP高油酸分子标记检测方法,优化了KASP分子标记检测体系,利用分子标记辅助连续回交,结合近红外品质快速检测技术及南繁加代技术,以河南省大面积推广的豫花15、远杂9102、豫花9327、豫花9326四个不同类型品种为轮回亲本, 5年内连续回交4代、自交4代,定向获得了4个轮回亲本遗传背景的BC4F4和BC4F5稳定高油酸改良材料24个。调查分析了BC4F4和BC4F5单株的13个农艺性状与轮回亲本的相似度,并利用轮回亲本与非轮回亲本之间的差异SNP的KASP分子标记进行了BC4F4和BC4F5株系的轮回亲本遗传背景检测。结果表明,轮回亲本的遗传背景在BC4F5的比例为79.49%～92.31%。本研究为快速高效改良花生油酸含量探索了新的方法,获得的新品系拓展了高油酸花生的遗传背景,获得的一系列近等基因系可作为遗传研究材料进一步加以利用。
High oleic acid content is a key quality trait in peanut varieties.Peanut and its products with high oleic acid content possess better quality stability and nutrition for benefiting peanut processing and human health.To date,the released high oleic acid varieties were few and most of which were derived by conventional breeding with narrow genetic background.In this study,Allele Specific-PCR-Mispairing(AS-PCR-MP) method and optimized KASP assay system were developed to facilitate successive marker-assisted backcross breeding strategy for high oleic acid content.We also applied Near Infra-Red technology and winter nursery in the five-year successive backcross and selfing.Four types of cultivars(Yuhua 15,Yuanza 9102,Yuhua 9327,and Yuhua9326) were selected as recurrent parents for backcrossing.Twenty-four BC4 F4 and BC4 F5 stable lines with different genetic backgrounds and high oleic acid content were produced from four generations of successive backcrosses and four generations of BC4 selfing.Similarities between BC4 selfing progenies and recurrent parents were analyzed based on 13 agronomic traits.The recovery rate of genetic background for BC4 F4 and BC4 F5 was investigated by designing KASP assays with SNPs that were polymorphic between recurrent parents and non-recurrent parents.The proportion of genetic background of recurrent parents in BC4 F5 was79.49% to 92.31%.This study provides a new strategy for efficiently improving oleic acid content of peanuts with diverse genetic backgrounds.The near isogenic lines obtained in this study could be valuable genetic resources for further utilizations.
High oleic acid content is a key quality trait in peanut varieties.Peanut and its products with high oleic acid content possess better quality stability and nutrition for benefiting peanut processing and human health.To date,the released high oleic acid varieties were few and most of which were derived by conventional breeding with narrow genetic background.In this study,Allele Specific-PCR-Mispairing(AS-PCR-MP) method and optimized KASP assay system were developed to facilitate successive marker-assisted backcross breeding strategy for high oleic acid content.We also applied Near Infra-Red technology and winter nursery in the five-year successive backcross and selfing.Four types of cultivars(Yuhua 15,Yuanza 9102,Yuhua 9327,and Yuhua9326) were selected as recurrent parents for backcrossing.Twenty-four BC4 F4 and BC4 F5 stable lines with different genetic backgrounds and high oleic acid content were produced from four generations of successive backcrosses and four generations of BC4 selfing.Similarities between BC4 selfing progenies and recurrent parents were analyzed based on 13 agronomic traits.The recovery rate of genetic background for BC4 F4 and BC4 F5 was investigated by designing KASP assays with SNPs that were polymorphic between recurrent parents and non-recurrent parents.The proportion of genetic background of recurrent parents in BC4 F5 was79.49% to 92.31%.This study provides a new strategy for efficiently improving oleic acid content of peanuts with diverse genetic backgrounds.The near isogenic lines obtained in this study could be valuable genetic resources for further utilizations.
引文
[1]O’Keefe S F,Knauft D A.Comparison of oxidative stability of high and normal-oleic peanut oils.J Am Oil Chem Soc,1993,70:489-492.
[2]Talcott S T,Pozo-Insfran D D,Gorbet D W.Polyphenolic and antioxidant changes during storage of normal,mid,and high oleic acid peanuts.Food Chem,2005,89:77-84.
[3]Talcott S T,Duncan C E,Gorbet D W.Polyphenolic content and sensory properties of normal and high oleic acid peanuts.Food Chem,2005,90:379-388.
[4]Vassiliou E K,Garcia C,Tadros J H,Chakraborty G,Toney J H.Oleic acid and peanut oil high in oleic acid reverse the inhibitory effect of insulin production of the inflammatory cytokine TNF-alpha both in vitro and in vivo systems.Lipids Health Dis,2009,8:25-34.
[5]Norden A J,Gorbet D W,Knauft D A,Young C T.Variability in oil quality among peanut genotypes in the Florida breeding program.Peanut Sci,1987,14:7-11.
[6]禹山林,Isleib T G.美国大花生脂肪酸的遗传分析.中国油料作物学报,2000,22:34-37.Yu S L,Isleib T G.The inheritance of high oleic acid content in peanut of Virginia type in USA.Chin J Oil Crop Sci,2000,22:34-37(in Chinese with English abstract).
[7]黄冰艳,张新友,苗利娟,刘华,秦利,徐静,张忠信,汤丰收,董文召,韩锁义,刘志勇.花生油酸和亚油酸含量的遗传模式分析.中国农业科学,2012,45:617-624.Huang B Y,Zhang X Y,Miao L J,Liu H,Qin L,Xu J,Zhang Z X,Tang F S,Dong W Z,Han S Y,Liu Z Y.Inheritance analysis of oleicacid and linoleic acid content of Arachis hypogaea L.Sci Agric Sin,2012,45:617-624(in Chinese with English abstract).
[8]Jung S,Swift D,Sengoku E,Patel M,Teule F,Powell G,Moore K,Abbott A.The high oleate trait in the cultivated peanut(Arachis hypogaea L.):I.Isolation and characterization of two genes encoding microsomal oleoyl-PC desaturases.Mol Gen Genet,2000,263:796-805.
[9]Jung S,Powell G,Moore K,Abbott A.The high oleate trait in the cultivated peanut(Arachis hypogaea L.):II.Molecular basis and genetics of the trait.Mol Gen Genet,2000,263:806-811.
[10]Lopez Y,Nadaf H L,Smith O D,Connell J P,Reddy A S,Fritz AK.Isolation and characterization of theΔ12-fatty acid desaturase in peanut(Arachis hypogaea L.)and search for polymorphisms for the high oleate trait in Spanish market-type lines.Theor Appl Genet,2000,101:1131-1138.
[11]Lopez Y,Smith O D,Senseman S A,Rooney W L.Genetic factors influencing high oleic acid content in Spanish market-type peanut cultivars.Crop Sci,2001,41:51-56.
[12]Bruner A C,Jung S,Abbott A G,Powell G L.The naturally occurring high oleate oil character in some peanut varieties results from reduced oleoyl-PC desaturase activity from mutation of aspartate 150 to Asparagine.Crop Sci,2001,41:522-526.
[13]Chu Y,Holbrook C C,Tillman B L,Person G,Ozias-Akins P.Marker-assisted selection to pyramid nematode resistance and the high oleic trait in peanut.Plant Genome J,2011,4:110-117.
[14]Barkley N A,Wang M L,Pittman R N.Development of a real-time PCR genotyping assay to identify high oleic acid peanuts(Arachis hypogaea L.).Mol Breed,2009,25:541-548.
[15]Chen Z,Wang M L,Barkley N A,Pittman R N.A simple allele-specific PCR assay for detecting FAD2 alleles in both Aand B genomes of the cultivated peanut for high-oleate trait selection.Plant Mol Biol Rep,2010,28:542-548.
[16]徐平丽,唐桂英,付春,刘玮,鲁成凯,姜言生,刘皓,单雷.高通量检测花生油酸含量相关基因AhFAD2等位变异的方法.农业生物技术学报,2016,24:1364-1373.Xu P L,Tang G Y,Fu C,Liu W,Lu C K,Jiang Y S,Liu H,Shan L.Methods for high-throughput detecting the allelic variation of AhFAD2 gene related with oleic acid content in peanut(Arachis hypogaea).J Agric Biotechnol,2016,24:1364-1373(in Chinese with English abstract).
[17]Zhao S Z,Li A,Li C,Xia H,Zhao C,Zhang Y,Hou L,Wang X.Development and application of KASP marker for high throughput detection of AhFAD2 mutation in peanut.Elect J Biotechnol,2017,25:9-12.
[18]陈静.高油酸花生遗传育种研究进展.植物遗传资源学报,2011,12:190-196.Chen J.Advances in genetics and breeding of high oleic acid peanut.J Plant Genet Resour,2011,12:190-196(in Chinese with English abstract).
[19]王传堂,朱立贵.高油酸花生.上海:上海科学技术出版社,2017.pp 189-241.Wang C T,Zhu L G.High Oleic Acid Peanut.Shanghai:Shanghai Scientific and Technical Publisher,2017.pp 189-241(in Chinese).
[20]禹山林.中国花生品种及其系谱.上海:上海科学技术出版社,2008.pp 322-345.Yu S L.Chinese Peanut Variety and Pedigree.Shanghai:Shanghai Scientific and Technical Publisher,2008.pp 322-345(in Chinese).
[21]Patel M,Jung S,Moore K,Powell G,Ainsworth C,Abbott A.High-oleate peanut mutants results from a MITE insertion into the FAD2 gene.Theor Appl Genet,2004,108:1492-1502.
[22]Trick M,Mugford S G,Jiang C C,Febrer M,Uauy C.Combining SNP discovery from next-generation sequencing data with bulked segregant analysis(BSA)to fine-map genes in polyploid wheat.BMC Plant Biol,2012,12:14.
[23]姜慧芳,段乃雄.花生种质资源描述规范和数据标准.北京:中国农业出版社,2006.pp 65-72.Jiang H F,Duan N X.Descriptors and Dada Standard for Peanut.Beijing:China Agriculture Press,2006.pp 65-72(in Chinese).
[24]姜慧芳,任小平,黄家权,廖伯寿,雷永.中国花生小核心种质的建立及高油酸基因源的发掘.中国油料作物学报,2008,30:294-299.Jiang H F,Ren X P,Huang J Q,Liao B S,Lei Y.Establishment of peanut mini core collection in China and exploration of new resource with high oleate.Chin J Oil Crop Sci,2008,30:294-299(in Chinese with English abstract).
[25]雷永,姜慧芳,文奇根,黄家权,晏立英,廖伯寿.ahFAD2A等位基因在中国花生小核心种质中的分布及其与种子油酸含量的相关性分析.作物学报,2010,36:1864-1869.Lei Y,Jiang H F,Wen Q G,Huang J Q,Yan L Y,Liao B S.Frequencies of ahFAD2A alleles in Chinese peanut mini core collection and its correlation with oleic acid content.Acta Agron Sin,2010,36:1864-1869(in Chinese with English abstract).
[26]黄冰艳,张新友,苗利娟,高伟,韩锁义,董文召,汤丰收,刘志勇.花生ahFAD2A等位基因表达变异与种子油酸积累关系.作物学报,2012,38:1752-1759.Huang B Y,Zhang X Y,Miao L J,Gao W,Han S Y,Dong W Z,Tang F S,Liu Z Y.Allelic expression variation of ahFAD2A and its relationship with oleic acid accumulation in peanut.Acta Agron Sin,2012,38:1752-1759(in Chinese with English abstract).
[27]黄冰艳,张新友,董文召,臧秀旺,苗利娟,刘华,高伟,韩锁义,汤丰收.河南省地方花生资源的品质性状及其育种利用途径.植物遗传资源学报,2012,13:414-417.Huang B Y,Zhang X Y,Dong W Z,Zang X W,Miao L J,Liu H,Gao W,Han S Y,Tang F S.Profiling of quality characteristics for peanut germplasm from Henan Province and its breeding strategy.J Plant Genet Resour,2012,13:414-417(in Chinese with English abstract).
[28]张照华,王志慧,淮东欣,谭家壮,陈剑洪,晏立英,王晓军,万丽云,陈傲,康彦平,姜慧芳,雷永,廖伯寿.利用回交和分子标记辅助选择快速培养高油酸花生品种及其评价.中国农业科学,2018,51:1641-1652.Zhang Z H,Wang Z H,Huai D X,Tan J Z,Chen J H,Yan L Y,Wang X J,Wan L Y,Chen A,Kang Y P,Jiang H F,Lei Y,Liao BS.Fast development of high oleate peanut cultivars by using marker-assisted backcrossing and their evaluation.Sci Agric Sin,2018,51:1641-1652(in Chinese with English abstract).
[29]于明洋,孙明明,郭悦,姜平平,雷永,黄冰艳,冯素萍,郭宝珠,隋炯明,王晶珊,乔利仙.利用回交法快速选育高油酸新品系.作物学报,2017,43:855-861.Yu M Y,Sun M M,Guo Y,Jiang P P,Lei Y,Huang B Y,Feng S P,Guo B Z,Sui J M,Wang J S,Qiao L X.Breeding new peanut lines with high oleic acid content using backcross method.Acta Agron Sin,2017,43:855-861(in Chinese with English abstract).
[30]赵术珍,侯蕾,李长生,赵传志,任丽,李爱芹,邓丽,夏晗,王兴军.分子标记辅助回交选育高油酸花生新种质.中国油料作物学报,2017,39:30-36.Zhao S Z,Hou L,Li C S,Zhao C Z,Ren L,Li A Q,Deng L,Xia H,Wang X J.Development of high oleic acid peanut from molecular marker assisted selection.Chin J Oil Crop Sci,2017,39:30-36(in Chinese with English abstract).
[31]Varshney R K.Exciting journey of 10 years from genomes to fields and markets:some success stories of genomics-assisted breeding in chickpea,pigeonpea and groundnut.Plant Sci,2016,242:98-107.
[32]Fox G.Near infrared reflectance as a rapid and inexpensive surrogate measure for fatty acid composition and oil content of peanut(Arachis hypogaea L.).J Near Infrared Spectrosc,2005,13:287-291.
[33]Janila P,Pandey M K,Shasidhar Y,Variath M T,Sriswathi M,Khera P,Manohar S S,Nagesh P,Vishwakarma M K,Mishra G P,Radhakrishnan T,Manivannan N,Dobariya K L,Vasanthi R P,Varshney R K.Molecular breeding for introgression of fatty acid desaturase mutant alleles(ahFAD2A and ahFAD2B)enhances oil quality in high and low oil containing peanut genotypes.Plant Sci,2016,242:203-213.
[2]Talcott S T,Pozo-Insfran D D,Gorbet D W.Polyphenolic and antioxidant changes during storage of normal,mid,and high oleic acid peanuts.Food Chem,2005,89:77-84.
[3]Talcott S T,Duncan C E,Gorbet D W.Polyphenolic content and sensory properties of normal and high oleic acid peanuts.Food Chem,2005,90:379-388.
[4]Vassiliou E K,Garcia C,Tadros J H,Chakraborty G,Toney J H.Oleic acid and peanut oil high in oleic acid reverse the inhibitory effect of insulin production of the inflammatory cytokine TNF-alpha both in vitro and in vivo systems.Lipids Health Dis,2009,8:25-34.
[5]Norden A J,Gorbet D W,Knauft D A,Young C T.Variability in oil quality among peanut genotypes in the Florida breeding program.Peanut Sci,1987,14:7-11.
[6]禹山林,Isleib T G.美国大花生脂肪酸的遗传分析.中国油料作物学报,2000,22:34-37.Yu S L,Isleib T G.The inheritance of high oleic acid content in peanut of Virginia type in USA.Chin J Oil Crop Sci,2000,22:34-37(in Chinese with English abstract).
[7]黄冰艳,张新友,苗利娟,刘华,秦利,徐静,张忠信,汤丰收,董文召,韩锁义,刘志勇.花生油酸和亚油酸含量的遗传模式分析.中国农业科学,2012,45:617-624.Huang B Y,Zhang X Y,Miao L J,Liu H,Qin L,Xu J,Zhang Z X,Tang F S,Dong W Z,Han S Y,Liu Z Y.Inheritance analysis of oleicacid and linoleic acid content of Arachis hypogaea L.Sci Agric Sin,2012,45:617-624(in Chinese with English abstract).
[8]Jung S,Swift D,Sengoku E,Patel M,Teule F,Powell G,Moore K,Abbott A.The high oleate trait in the cultivated peanut(Arachis hypogaea L.):I.Isolation and characterization of two genes encoding microsomal oleoyl-PC desaturases.Mol Gen Genet,2000,263:796-805.
[9]Jung S,Powell G,Moore K,Abbott A.The high oleate trait in the cultivated peanut(Arachis hypogaea L.):II.Molecular basis and genetics of the trait.Mol Gen Genet,2000,263:806-811.
[10]Lopez Y,Nadaf H L,Smith O D,Connell J P,Reddy A S,Fritz AK.Isolation and characterization of theΔ12-fatty acid desaturase in peanut(Arachis hypogaea L.)and search for polymorphisms for the high oleate trait in Spanish market-type lines.Theor Appl Genet,2000,101:1131-1138.
[11]Lopez Y,Smith O D,Senseman S A,Rooney W L.Genetic factors influencing high oleic acid content in Spanish market-type peanut cultivars.Crop Sci,2001,41:51-56.
[12]Bruner A C,Jung S,Abbott A G,Powell G L.The naturally occurring high oleate oil character in some peanut varieties results from reduced oleoyl-PC desaturase activity from mutation of aspartate 150 to Asparagine.Crop Sci,2001,41:522-526.
[13]Chu Y,Holbrook C C,Tillman B L,Person G,Ozias-Akins P.Marker-assisted selection to pyramid nematode resistance and the high oleic trait in peanut.Plant Genome J,2011,4:110-117.
[14]Barkley N A,Wang M L,Pittman R N.Development of a real-time PCR genotyping assay to identify high oleic acid peanuts(Arachis hypogaea L.).Mol Breed,2009,25:541-548.
[15]Chen Z,Wang M L,Barkley N A,Pittman R N.A simple allele-specific PCR assay for detecting FAD2 alleles in both Aand B genomes of the cultivated peanut for high-oleate trait selection.Plant Mol Biol Rep,2010,28:542-548.
[16]徐平丽,唐桂英,付春,刘玮,鲁成凯,姜言生,刘皓,单雷.高通量检测花生油酸含量相关基因AhFAD2等位变异的方法.农业生物技术学报,2016,24:1364-1373.Xu P L,Tang G Y,Fu C,Liu W,Lu C K,Jiang Y S,Liu H,Shan L.Methods for high-throughput detecting the allelic variation of AhFAD2 gene related with oleic acid content in peanut(Arachis hypogaea).J Agric Biotechnol,2016,24:1364-1373(in Chinese with English abstract).
[17]Zhao S Z,Li A,Li C,Xia H,Zhao C,Zhang Y,Hou L,Wang X.Development and application of KASP marker for high throughput detection of AhFAD2 mutation in peanut.Elect J Biotechnol,2017,25:9-12.
[18]陈静.高油酸花生遗传育种研究进展.植物遗传资源学报,2011,12:190-196.Chen J.Advances in genetics and breeding of high oleic acid peanut.J Plant Genet Resour,2011,12:190-196(in Chinese with English abstract).
[19]王传堂,朱立贵.高油酸花生.上海:上海科学技术出版社,2017.pp 189-241.Wang C T,Zhu L G.High Oleic Acid Peanut.Shanghai:Shanghai Scientific and Technical Publisher,2017.pp 189-241(in Chinese).
[20]禹山林.中国花生品种及其系谱.上海:上海科学技术出版社,2008.pp 322-345.Yu S L.Chinese Peanut Variety and Pedigree.Shanghai:Shanghai Scientific and Technical Publisher,2008.pp 322-345(in Chinese).
[21]Patel M,Jung S,Moore K,Powell G,Ainsworth C,Abbott A.High-oleate peanut mutants results from a MITE insertion into the FAD2 gene.Theor Appl Genet,2004,108:1492-1502.
[22]Trick M,Mugford S G,Jiang C C,Febrer M,Uauy C.Combining SNP discovery from next-generation sequencing data with bulked segregant analysis(BSA)to fine-map genes in polyploid wheat.BMC Plant Biol,2012,12:14.
[23]姜慧芳,段乃雄.花生种质资源描述规范和数据标准.北京:中国农业出版社,2006.pp 65-72.Jiang H F,Duan N X.Descriptors and Dada Standard for Peanut.Beijing:China Agriculture Press,2006.pp 65-72(in Chinese).
[24]姜慧芳,任小平,黄家权,廖伯寿,雷永.中国花生小核心种质的建立及高油酸基因源的发掘.中国油料作物学报,2008,30:294-299.Jiang H F,Ren X P,Huang J Q,Liao B S,Lei Y.Establishment of peanut mini core collection in China and exploration of new resource with high oleate.Chin J Oil Crop Sci,2008,30:294-299(in Chinese with English abstract).
[25]雷永,姜慧芳,文奇根,黄家权,晏立英,廖伯寿.ahFAD2A等位基因在中国花生小核心种质中的分布及其与种子油酸含量的相关性分析.作物学报,2010,36:1864-1869.Lei Y,Jiang H F,Wen Q G,Huang J Q,Yan L Y,Liao B S.Frequencies of ahFAD2A alleles in Chinese peanut mini core collection and its correlation with oleic acid content.Acta Agron Sin,2010,36:1864-1869(in Chinese with English abstract).
[26]黄冰艳,张新友,苗利娟,高伟,韩锁义,董文召,汤丰收,刘志勇.花生ahFAD2A等位基因表达变异与种子油酸积累关系.作物学报,2012,38:1752-1759.Huang B Y,Zhang X Y,Miao L J,Gao W,Han S Y,Dong W Z,Tang F S,Liu Z Y.Allelic expression variation of ahFAD2A and its relationship with oleic acid accumulation in peanut.Acta Agron Sin,2012,38:1752-1759(in Chinese with English abstract).
[27]黄冰艳,张新友,董文召,臧秀旺,苗利娟,刘华,高伟,韩锁义,汤丰收.河南省地方花生资源的品质性状及其育种利用途径.植物遗传资源学报,2012,13:414-417.Huang B Y,Zhang X Y,Dong W Z,Zang X W,Miao L J,Liu H,Gao W,Han S Y,Tang F S.Profiling of quality characteristics for peanut germplasm from Henan Province and its breeding strategy.J Plant Genet Resour,2012,13:414-417(in Chinese with English abstract).
[28]张照华,王志慧,淮东欣,谭家壮,陈剑洪,晏立英,王晓军,万丽云,陈傲,康彦平,姜慧芳,雷永,廖伯寿.利用回交和分子标记辅助选择快速培养高油酸花生品种及其评价.中国农业科学,2018,51:1641-1652.Zhang Z H,Wang Z H,Huai D X,Tan J Z,Chen J H,Yan L Y,Wang X J,Wan L Y,Chen A,Kang Y P,Jiang H F,Lei Y,Liao BS.Fast development of high oleate peanut cultivars by using marker-assisted backcrossing and their evaluation.Sci Agric Sin,2018,51:1641-1652(in Chinese with English abstract).
[29]于明洋,孙明明,郭悦,姜平平,雷永,黄冰艳,冯素萍,郭宝珠,隋炯明,王晶珊,乔利仙.利用回交法快速选育高油酸新品系.作物学报,2017,43:855-861.Yu M Y,Sun M M,Guo Y,Jiang P P,Lei Y,Huang B Y,Feng S P,Guo B Z,Sui J M,Wang J S,Qiao L X.Breeding new peanut lines with high oleic acid content using backcross method.Acta Agron Sin,2017,43:855-861(in Chinese with English abstract).
[30]赵术珍,侯蕾,李长生,赵传志,任丽,李爱芹,邓丽,夏晗,王兴军.分子标记辅助回交选育高油酸花生新种质.中国油料作物学报,2017,39:30-36.Zhao S Z,Hou L,Li C S,Zhao C Z,Ren L,Li A Q,Deng L,Xia H,Wang X J.Development of high oleic acid peanut from molecular marker assisted selection.Chin J Oil Crop Sci,2017,39:30-36(in Chinese with English abstract).
[31]Varshney R K.Exciting journey of 10 years from genomes to fields and markets:some success stories of genomics-assisted breeding in chickpea,pigeonpea and groundnut.Plant Sci,2016,242:98-107.
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