糜子EMS突变体库构建和突变体筛选
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摘要
糜子(Panicum miliaceum L.)具有抗旱、耐瘠、适应性广、生育期短等特点,是我国北方地区的杂粮作物。糜子高产品种不耐水肥、易倒伏,锄草、间苗费时费工的问题严重影响着农民对糜子生产的积极性和收入。EMS(甲基磺酸乙酯)化学诱变方法成本低,操作简单,诱变率高,是作物育种的重要方法。本试验选用糜子品种伊选大红糜进行EMS化学诱变而构建突变体库,对获得的2333个M2材料进行突变频率和全生育期表型多样性等特征的分析。结果显示,所构建的糜子突变体库材料变异类型非常丰富,共发现104个形态性状突变株系,突变频率为4.5%,包括叶色、叶型、株高、生育期、育性、穗型、种皮颜色等性状,该突变体库的构建将为糜子功能基因组学研究和育种提供丰富的突变体资源和新型种质。
Broomcorn millet(Panicum miliaceum L.)is one of the minor crops in China,which represented resistant and tolerant against drought,lower-fertilizer and abiotic stresses as well as short life-cycle. The problem of its easy to lodging characteristics,water and fertilizer intolerance of high-yield varieties,artificial thinning and weeding which need consuming a lot of labors and time have been become the bottlenecks of the production of broomcorn millet. The EMS chemical mutagenesis,which is low cost,simple in operation,and high in mutant rate,is an important method for crop breeding. Within this study,by generating an EMS-triggered mutant population by using broomcorn millet variety Yi-Xuan-Da-Hong-Mi,we analyzed the mutation frequency and phenotypic variation of 2333 M2 plants.Out of that,a total of 104 plants with visible phenotype differing with the parental line were identified,yielding 4. 5%of mutation rate. These mutants showed modifications on any of leaf color,leaf shape,plant height,mature period,fertility,spike shape,seed-coat color and so on. Thus,these mutants would become a genetic resource being useful in deciphering the gene function and breeding for new varieties in Broomcorn millet.
Broomcorn millet(Panicum miliaceum L.)is one of the minor crops in China,which represented resistant and tolerant against drought,lower-fertilizer and abiotic stresses as well as short life-cycle. The problem of its easy to lodging characteristics,water and fertilizer intolerance of high-yield varieties,artificial thinning and weeding which need consuming a lot of labors and time have been become the bottlenecks of the production of broomcorn millet. The EMS chemical mutagenesis,which is low cost,simple in operation,and high in mutant rate,is an important method for crop breeding. Within this study,by generating an EMS-triggered mutant population by using broomcorn millet variety Yi-Xuan-Da-Hong-Mi,we analyzed the mutation frequency and phenotypic variation of 2333 M2 plants.Out of that,a total of 104 plants with visible phenotype differing with the parental line were identified,yielding 4. 5%of mutation rate. These mutants showed modifications on any of leaf color,leaf shape,plant height,mature period,fertility,spike shape,seed-coat color and so on. Thus,these mutants would become a genetic resource being useful in deciphering the gene function and breeding for new varieties in Broomcorn millet.
引文
[1]林凡云,胡银岗,宋国琦,何蓓如.糜子抗旱及复水相关基因的cDNA-AFLP差异显示.干旱地区农业研究,2006,24(3):95-99Lin F Y,Hu Y G,Song G Q,He B R.Analysis on the responsive genes during drought and rehydration in broomcorn millet(Panicum miliaceum L.)by means of cdna-aflp.Agricultural Research in the Arid Areas,2006,24(3):95-99
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[4]Park M Y,Seo D W,Lee J Y,Jang H H,Park D S.Effects of Panicum miliaceum L.extract on adipogenic transcription factors and fatty acid accumulation in 3T3-L1 adipocytes.Nutrition Research&Practice,2011,5(3):192-197
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[8]李秀兰,孙小秋,王平荣,周慧,邓晓建.一个新的水稻黄绿叶突变体的遗传分析与基因定位.作物学报,2010,36(6):1050-1054Li X L,Sun X Q,Wang P R,Zhou H,Deng X J.Genetic analysis and gene mapping of a novel yellow-green leaf mutant in rice.Acta Agronomica Sinica,2010,36(6):1050-1054
[9]霍朋杰,吴斌,张宗文.裸燕麦EMS突变体库筛选与分析.植物遗传资源学报,2015,16(2):379-384Huo P J,Wu B,Zhang Z W.Identification and analysis of EMSmutant library in oat(Avena nuda L.).Journal of Plant Genetic Resources,2015,16(2):379-384
[10]韩锁义,张恒友,杨玛丽,赵团结,盖钧镒,喻德跃.大豆“南农86-4”突变体筛选及突变体库的构建.作物学报,2007,33(12):2059-2062Han S Y,Zhang H Y,Yang M L,Zhao T J,Gai J Y,Yu DY.Screening of mutants and construction of mutant population in soybean"nannong 86-4".Acta Agronomica Sinica,2007,33(12):2059-2062
[11]杨峰,刘威,吴楠,隆艳喜,任旭东,聂丹丹,陈亮,王宁宁,马建.EMS诱导水稻一目惚(Hitomebore)表型和基因组序列变异.分子植物育种,2017,15(10):4080-4085Yang F,Liu W,Wu N,Long Y X,Ren X D,Nie D D,Chen L,Wang N N,Ma J.EMS induced transposition of maping transposons in hitomebore genome of rice.Molecular Plant Breeding,2017,15(10):4080-4085
[12]Roswitha B,Axel P,Günter S,Karlheinz S.Development of a TILLING resource in durum wheat for reverse-and forwardgenetic analyses.Crop&Pasture Science,2014,65(1):112-124
[13]冯学金,郭秀娟,杨建春,王利琴.诱变技术在亚麻育种中的应用.核农学报,2017,31(7):1310-1316Feng X J,Guo X J,Yang J C,Wang L Q.Application of mutation technology in flax breeding.Journal of Nuclear Agricultural Sciences,2017,31(7):1310-1316
[14]张以忠,邓琳琼.EMS诱变苦荞突变体与其亲本的比较分析.热带作物学报,2017,38(9):1601-1606Zhang Y Z,Deng L Q.Comparative analysis of fagopyrum tataricum mutants and its parents in EMS mutagenesis.Chinese Journal of Tropical Crops,2017,38(9):1601-1606
[15]Jiao Y,Burke J J,Chopra R,Burow G,Chen J,Wang B,Hayes C,Emendack Y,Ware D,Xin Z.A sorghum mutant resource as an efficient platform for gene discovery in grasses.Plant Cell,2016,28(7):1551-1562
[16]Jung K H,Hur J,Ryu C H,Choi Y J,Chung Y Y,Miyao A,Hirochika H,An G.Characterization of a rice chlorophylldeficient mutant using the T-DNA gene-trap system.Plant&Cell Physiology,2003,44(5):463-472
[17]Motohashi R,Ito T,Kobayashi M,Taji T,Nagata N,Asami T,Shinozaki K.Functional analysis of the 37kDa inner envelope membrane polypeptide in chloroplast biogenesis using a Dstagged Arabidopsis,pale-green mutant.Plant Journal for Cell&Molecular Biology,2003,34(5):719-731
[18]董凤高,朱旭东,熊振民,程式华,孙宗修,闵绍楷.以淡绿叶为标记的籼型光-温敏核不育系M2S的选育.中国水稻科学,1995,9(2):65-70Dong F G,Zhu X D,Xiong Z M,Cheng S H,Sun Z X,Min SK.Breeding of a photo-thermoperiod sensitive genie male sterile indica rice with a pale-green-leaf marker.Chinese Journal of Rice Science,1995,9(2):65-70
[19]李素芳,成浩,陈明,虞富莲,晏静,刘益民,陈龙安.安吉白茶特异性状的生理生化本质.茶叶科学,1999,19(2):87-92Li S F,Cheng H,Chen M,Yu F L,Yan J,Liu Y M,Chen LA.Physiological and biochemical essence of the extraordinary characters of Anji Baicha.Journal of Tea Science,1999,19(2):87-92
[20]刁现民,程汝宏.“谷子糜子育种”专题导读:十五年区试数据分析展示谷子糜子育种现状.中国农业科学,2017,50(23):4469-4474Diao X M,Cheng R H.Current breeding situation of foxtail millet and common millet in china as revealed by exploitation of 15 years regional adaptation test data.Scientia Agricultura Sinica,2017,50(23):4469-4474
[21]杨清华,邱军,李海,杨天育,程炳文,赵敏,刘国庆,高小丽,冯佰利.糜子育成品种农艺、产量及品质性状综合鉴定与评价.中国农业科学,2017,50(23):4530-4544Yang Q H,Qiu J,Li H,Yang T Y,Cheng B W,Zhao M,Liu G Q,Gao X L,Feng B L,Comprehensive evaluation of agronomic,yield and quality traits of broomcorn millet(Panicum miliaceum L.)cultivars.Scientia Agricultura Sinica,2017,50(23):4530-4544
[22]董孔军,刘天鹏,何继红,任瑞玉,张磊,杨天育.糜子种质材料的抗倒伏性、农艺性状及力学特性.西北农业学报,2018,27(8):1119-1126Dong K J,Liu T P,He J H,Ren R Y,Zhang L,Yang TY.Interrelation of lodging resistance,agronomic traits and mechanical properties of germplasm resources in broomcorn millet.Acta Agriculturae Boreali-occidentalis Cinica,2018,27(8):1119-1126
[23]Huang X Z,Qian Q,Liu Z B,Sun H Y,He S Y,Luo D,Xia G M,Chu C G,Li J Y,Fu X D.Natural variation at the DEP1locus enhances grain yield in rice.Nature Genetics,2009,41(4):494-497
[24]Li F,Liu W B,Tang J Y,Chen J F,Tong H G,Hu B,Li CL,Fang J,Chen M S,Chu C C.Rice DENSE AND ERECTPANICLE 2 is essential for determining panicle outgrowth and elongation.Cell Research,2010,20(7):838-849
[25]Boss P K,Bastow R M,Mylne J S,Dean C.Multiple pathways in the decision to flower:enabling,promoting,and resetting.Plant Cell,2004,16(Suppl):S18-S31
[2]柴岩.糜子(黄米)的营养和生产概况.粮食加工,2009,34(4):90-91Chai Y.Nutrition and production survey of broomcorn millet.Grain Processing,2009,34(4):90-91
[3]Kulp K.Handbook of Cereal Science and Technology,Second Edition.,Manhattan,Kansas:CRC Press,2000:1-3
[4]Park M Y,Seo D W,Lee J Y,Jang H H,Park D S.Effects of Panicum miliaceum L.extract on adipogenic transcription factors and fatty acid accumulation in 3T3-L1 adipocytes.Nutrition Research&Practice,2011,5(3):192-197
[5]Glaszmann J C,Kilian B,Upadhyaya H D,Varshney RK.Accessing genetic diversity for crop improvement.Current Opinion in Plant Biology,2010,13(2):167-173
[6]Greene E A,Codomo C A,Taylor N E,Henikoff J G,Till B J,Reynolds S H,Comai L.Spectrum of chemically induced mutations from a large-scale reverse-genetic screen in Arabidopsis.Genetics,2003,164(2):731-740
[7]李韬,骆孟,钱丹,董晶晶,顾世梁.抗赤霉病小麦地方品种黄方柱和海盐种EMS突变体的变异分析.植物遗传资源学报,2016,17(6):1092-1098Li T,Luo M,Qian D,Dong J J,Gu S L.Evaluation of phenotypic variation of ems-induced mutants of Chinese wheat landraces Huangfangzhu and Haiyanzhong resistant to fusarium head blight.Journal of Plant Genetic Resources,2016,17(6):1092-1098
[8]李秀兰,孙小秋,王平荣,周慧,邓晓建.一个新的水稻黄绿叶突变体的遗传分析与基因定位.作物学报,2010,36(6):1050-1054Li X L,Sun X Q,Wang P R,Zhou H,Deng X J.Genetic analysis and gene mapping of a novel yellow-green leaf mutant in rice.Acta Agronomica Sinica,2010,36(6):1050-1054
[9]霍朋杰,吴斌,张宗文.裸燕麦EMS突变体库筛选与分析.植物遗传资源学报,2015,16(2):379-384Huo P J,Wu B,Zhang Z W.Identification and analysis of EMSmutant library in oat(Avena nuda L.).Journal of Plant Genetic Resources,2015,16(2):379-384
[10]韩锁义,张恒友,杨玛丽,赵团结,盖钧镒,喻德跃.大豆“南农86-4”突变体筛选及突变体库的构建.作物学报,2007,33(12):2059-2062Han S Y,Zhang H Y,Yang M L,Zhao T J,Gai J Y,Yu DY.Screening of mutants and construction of mutant population in soybean"nannong 86-4".Acta Agronomica Sinica,2007,33(12):2059-2062
[11]杨峰,刘威,吴楠,隆艳喜,任旭东,聂丹丹,陈亮,王宁宁,马建.EMS诱导水稻一目惚(Hitomebore)表型和基因组序列变异.分子植物育种,2017,15(10):4080-4085Yang F,Liu W,Wu N,Long Y X,Ren X D,Nie D D,Chen L,Wang N N,Ma J.EMS induced transposition of maping transposons in hitomebore genome of rice.Molecular Plant Breeding,2017,15(10):4080-4085
[12]Roswitha B,Axel P,Günter S,Karlheinz S.Development of a TILLING resource in durum wheat for reverse-and forwardgenetic analyses.Crop&Pasture Science,2014,65(1):112-124
[13]冯学金,郭秀娟,杨建春,王利琴.诱变技术在亚麻育种中的应用.核农学报,2017,31(7):1310-1316Feng X J,Guo X J,Yang J C,Wang L Q.Application of mutation technology in flax breeding.Journal of Nuclear Agricultural Sciences,2017,31(7):1310-1316
[14]张以忠,邓琳琼.EMS诱变苦荞突变体与其亲本的比较分析.热带作物学报,2017,38(9):1601-1606Zhang Y Z,Deng L Q.Comparative analysis of fagopyrum tataricum mutants and its parents in EMS mutagenesis.Chinese Journal of Tropical Crops,2017,38(9):1601-1606
[15]Jiao Y,Burke J J,Chopra R,Burow G,Chen J,Wang B,Hayes C,Emendack Y,Ware D,Xin Z.A sorghum mutant resource as an efficient platform for gene discovery in grasses.Plant Cell,2016,28(7):1551-1562
[16]Jung K H,Hur J,Ryu C H,Choi Y J,Chung Y Y,Miyao A,Hirochika H,An G.Characterization of a rice chlorophylldeficient mutant using the T-DNA gene-trap system.Plant&Cell Physiology,2003,44(5):463-472
[17]Motohashi R,Ito T,Kobayashi M,Taji T,Nagata N,Asami T,Shinozaki K.Functional analysis of the 37kDa inner envelope membrane polypeptide in chloroplast biogenesis using a Dstagged Arabidopsis,pale-green mutant.Plant Journal for Cell&Molecular Biology,2003,34(5):719-731
[18]董凤高,朱旭东,熊振民,程式华,孙宗修,闵绍楷.以淡绿叶为标记的籼型光-温敏核不育系M2S的选育.中国水稻科学,1995,9(2):65-70Dong F G,Zhu X D,Xiong Z M,Cheng S H,Sun Z X,Min SK.Breeding of a photo-thermoperiod sensitive genie male sterile indica rice with a pale-green-leaf marker.Chinese Journal of Rice Science,1995,9(2):65-70
[19]李素芳,成浩,陈明,虞富莲,晏静,刘益民,陈龙安.安吉白茶特异性状的生理生化本质.茶叶科学,1999,19(2):87-92Li S F,Cheng H,Chen M,Yu F L,Yan J,Liu Y M,Chen LA.Physiological and biochemical essence of the extraordinary characters of Anji Baicha.Journal of Tea Science,1999,19(2):87-92
[20]刁现民,程汝宏.“谷子糜子育种”专题导读:十五年区试数据分析展示谷子糜子育种现状.中国农业科学,2017,50(23):4469-4474Diao X M,Cheng R H.Current breeding situation of foxtail millet and common millet in china as revealed by exploitation of 15 years regional adaptation test data.Scientia Agricultura Sinica,2017,50(23):4469-4474
[21]杨清华,邱军,李海,杨天育,程炳文,赵敏,刘国庆,高小丽,冯佰利.糜子育成品种农艺、产量及品质性状综合鉴定与评价.中国农业科学,2017,50(23):4530-4544Yang Q H,Qiu J,Li H,Yang T Y,Cheng B W,Zhao M,Liu G Q,Gao X L,Feng B L,Comprehensive evaluation of agronomic,yield and quality traits of broomcorn millet(Panicum miliaceum L.)cultivars.Scientia Agricultura Sinica,2017,50(23):4530-4544
[22]董孔军,刘天鹏,何继红,任瑞玉,张磊,杨天育.糜子种质材料的抗倒伏性、农艺性状及力学特性.西北农业学报,2018,27(8):1119-1126Dong K J,Liu T P,He J H,Ren R Y,Zhang L,Yang TY.Interrelation of lodging resistance,agronomic traits and mechanical properties of germplasm resources in broomcorn millet.Acta Agriculturae Boreali-occidentalis Cinica,2018,27(8):1119-1126
[23]Huang X Z,Qian Q,Liu Z B,Sun H Y,He S Y,Luo D,Xia G M,Chu C G,Li J Y,Fu X D.Natural variation at the DEP1locus enhances grain yield in rice.Nature Genetics,2009,41(4):494-497
[24]Li F,Liu W B,Tang J Y,Chen J F,Tong H G,Hu B,Li CL,Fang J,Chen M S,Chu C C.Rice DENSE AND ERECTPANICLE 2 is essential for determining panicle outgrowth and elongation.Cell Research,2010,20(7):838-849
[25]Boss P K,Bastow R M,Mylne J S,Dean C.Multiple pathways in the decision to flower:enabling,promoting,and resetting.Plant Cell,2004,16(Suppl):S18-S31