离体诱变定向培育高油花生新品种宇花9号
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摘要
笔者前期研究发现经干旱胁迫处理后的花生叶片水势与其籽仁含油率呈显著正相关。文中对羟脯氨酸作为水势(渗透压)调节物质用于离体定向筛选花生高油突变体及培育花生新品种进行了研究。以花生品种花育20号胚小叶作为外植体,平阳霉素作为诱变剂添加于体胚诱导培养基上进行离体诱变培养。形成的体胚转移到添加6 mmol/L羟脯氨酸(培养基水势为-2.079 MPa)的体胚萌发和再生培养基上诱导体胚萌发成苗,同时进行高油突变体定向筛选。再生小苗经嫁接移栽田间,从再生植株后代中获得了132份含油率55%以上的高油突变体,其中27份含油率超过58%,2份超过60%。再生植株后代结合系谱育种法育成了高产高油花生新品种宇花9号,在辽宁省花生新品种备案试验中,籽仁产量比对照品种增产14.0%,并通过了国家非主要农作物品种登记。宇花9号含油率达61.05%,比亲本花育20号高11.55个百分点,是目前国际上含油率最高的花生品种。本研究结果表明,利用离体诱变、培养基低水势定向筛选及其再生植株后代结合常规育种法选择是定向培育高油花生品种的有效方法。
Leaf water potential of peanut subjected to drought stress is positively related to the oil content of peanut kernels.The aim of this study was to directly screen the high oil mutants of peanut and create the new peanut varieties using hydroxyproline as water potential regulator.In vitro mutagenesis was carried out with the embryonic leaflets of peanut variety Huayu 20 as explants and pingyangmycin as a mutagen added into the somatic embryo formation medium.The formed somatic embryos were successively transferred to somatic embryo germination and selection medium containing 6 mmol/L hydroxy pro line(at-2.079 MPa water potential)to induce regeneration and directionally screen high oil content mutants.After that,these plantlets were grafted and transplanted to the experimental field and 132 high oil mutants with oil content over 55% were obtained from the offspring of regenerated plants.Finally,among them,the oil contents of 27 lines were higher than 58% and of 2 lines were higher than 60%.A new peanut variety Yuhua 9 with high yield and oil content was bred from the regenerated plant progenies combining the pedigree breeding method.The yield was 14.0% higher than that of the control cultivar in the testing new peanut varieties of Liaoning province,and also it has passed the national registration of non-major crop varieties.Yuhua 9 with an oil content of 61.05%,which was 11.55 percentage points higher than that of the parent Huayu20,was the peanut cultivar with the highest oil content in the world.The result showed that it was an effective way for directional breeding of high oil peanut varieties by means of the three-step technique including in vitro mutagenesis,directional screening by reducing water potential in medium and pedigree selection of regenerated plant progenies.
Leaf water potential of peanut subjected to drought stress is positively related to the oil content of peanut kernels.The aim of this study was to directly screen the high oil mutants of peanut and create the new peanut varieties using hydroxyproline as water potential regulator.In vitro mutagenesis was carried out with the embryonic leaflets of peanut variety Huayu 20 as explants and pingyangmycin as a mutagen added into the somatic embryo formation medium.The formed somatic embryos were successively transferred to somatic embryo germination and selection medium containing 6 mmol/L hydroxy pro line(at-2.079 MPa water potential)to induce regeneration and directionally screen high oil content mutants.After that,these plantlets were grafted and transplanted to the experimental field and 132 high oil mutants with oil content over 55% were obtained from the offspring of regenerated plants.Finally,among them,the oil contents of 27 lines were higher than 58% and of 2 lines were higher than 60%.A new peanut variety Yuhua 9 with high yield and oil content was bred from the regenerated plant progenies combining the pedigree breeding method.The yield was 14.0% higher than that of the control cultivar in the testing new peanut varieties of Liaoning province,and also it has passed the national registration of non-major crop varieties.Yuhua 9 with an oil content of 61.05%,which was 11.55 percentage points higher than that of the parent Huayu20,was the peanut cultivar with the highest oil content in the world.The result showed that it was an effective way for directional breeding of high oil peanut varieties by means of the three-step technique including in vitro mutagenesis,directional screening by reducing water potential in medium and pedigree selection of regenerated plant progenies.
引文
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[8]Yu XL,Liu LX,Qiao LX,et al.Effects of mixed high energy particle field on embryonic leaflet culture and plant regeneration of peanut.J Nucl Agric Sci,2012,26(3):433-438(in Chinese).于新玲,刘录祥,乔利仙,等.高能混合粒子场辐照对花生胚小叶组织培养及植株再生的影响.核农学报,2012,26(3):433-438.
[9]Wang JS,Sui JM,Xie YD,et al.Generation of peanut mutants by fast neutron irradiation combined with in vitro culture.J Radiat Res,2015,56(3):437-445.
[10]Wang JS,Zhao MX,Qiao LX,et al.Effects of fast neutron irradiation on plant regeneration in embryonic leaflet culture of peanut(Arachis hypogaea L.).Chin JOil Crop Sci,2013,35(2):148-152(in Chinese).王晶珊,赵明霞,乔利仙,等.快中子辐照对花生种子胚小叶植株再生的影响.中国油料作物学报,2013,35(2):148-152.
[11]Wang JS,Qiao LX,Zhao LS,et al.Performance of peanut mutants and their offspring generated from mixed high-energy particle field radiation and tissue culture.Genet Mol Res,2015,14(3):10837-10848.
[12]Wang Y,Qiao LX,Wu XL,et al.Effect of in vitro mutagenesis with pingyangmycin and NaCl-directed screening on production and quality of offspring in peanut.Acta Agric Boreali-Sin,2015,30(1):202-206(in Chinese).王亚,乔利仙,武秀玲,等.平阳霉素诱变与NaCl定向筛选对花生后代产量和品质性状的影响.华北农学报,2015,30(1):202-206.
[13]Rai MK,Kalia RK,Singh R,et al.Developing stress tolerant plants through in vitro selection-an overview of the recent progress.Environ Exp Bot,2011,71(1):89-98.
[14]Wu WG,Liu GR,Yang XJ.Applications of the mutation in connection with in vitro culture for plant breeding.Chin Agric Sci Bull,2005,21(11):197-201(in Chinese).吴伟刚,刘桂茹,杨学举.诱变与组织培养相结合在植物育种中的应用.中国农学通报,2005,21(11):197-201.
[15]Patade VY,Suprasanna P.An in vitro radiation induced mutagenesis-selection system for salinity tolerance in sugarcane.Sugar Tech,2009,11(3):246-251.
[16]Luo J,Zhou HC,Wang YQ,et al.EMS mutagenesis in vitro and screening of calli from strawberry leaves of resistance to Botrytis chinerea Pers.J Nucl Agric Sci,2009,23(1):90-94(in Chinese).罗静,周厚成,王永清,等.EMS离体诱变及抗草莓灰霉病愈伤组织的筛选.核农学报,2009,23(1):90-94.
[17]Chen L,Dong JW,Tang Y,et al.Selection of salt-tolerant mutants from ethyl methanesulfonatemutagenized poplar embryonic calli.Acta Agric Shanghai,2007,23(3):86-91(in Chinese).陈丽,董举文,唐寅,等.EMS诱变处理定向筛选杨树耐盐突变体研究.上海农业学报,2007,23(3):86-91.
[18]Li H,Li B,Zhao HB,et al.Study on alkalinity resistance of alfalfa callus in mutagenic treatments.Pratac Sci,2009,26(7):32-35(in Chinese).李红,李波,赵洪波,等.诱变处理苜蓿愈伤组织抗碱性的研究.草业科学,2009,26(7):32-35.
[19]Zhao MX,Sun HY,Ji RR,et al.In vitro mutagenesis and directed screening for salt-tolerant mutants in peanut.Euphytica,2013,193(1):89-99.
[20]Wang X,Li Y,Zhu H,et al.Directional screening on peanut mutant with high oil content and correlation between oil content and leaf water potential.J Qingdao Agric Univ,2019,36(1):30-33(in Chinese).王霞,李艳,朱虹,等.花生高油突变体的定向筛选及籽仁含油率与叶片水势的相关性分析.青岛农业大学学报,2019,36(1):30-33.
[21]Zhang DD,Wang WH,Li G,et al.Chemistry mutagenesis in vitro and characteristics of mutants in sweet potato.Plant Physiol J,2016,52(3):343-348(in Chinese).张丹丹,王维华,李冠,等.甘薯化学离体诱变及突变体的性状表现.植物生理学报,2016,52(3):343-348.
[22]Yan ZM,Feng YN,Han YL,et al.Effects of exogenous proline on proline metabolism of Cucumis melo under salt stress.Acta Bot Bor Occid Sin,2015,35(10):2035-2041(in Chinese).颜志明,冯英娜,韩艳丽,等.外源脯氨酸对盐胁迫下甜瓜脯氨酸代谢的影响.西北植物学报,2015,35(10):2035-2041.
[23]Gillman JD,Tetlow A,Hagely K,et al.Identification of the molecular genetic basis of the low palmitic acid seed oil trait in soybean mutant line rg3 and association analysis of molecular markers with elevated seed stearic acid and reduced seed palmitic acid.Mol Breed,2014,34(2):447-455.
[24]Zhang YB,Xiao L,Dong C,et al.Comparative study on mutagenic effects of pingyangmycin and NaN3 on wheat.J Henan Instit Sci Technol:Nat Sci Ed,2015,43(5):6-9(in Chinese).张彦波,肖磊,董策,等.平阳霉素和NaN3对小麦诱变效应的比较研究.河南科技学院学报:自然科学版,2015,43(5):6-9.
[25]Zeng XH.Comparing effectiveness of different mutagens for seed quality and analysis of mutants in Brassica napus[D].Wuhan:Huazhong Agricultural University,2010(in Chinese).曾新华.不同诱变方法对油菜种子诱变效果及突变体的研究[D].武汉:华中农业大学,2010.
[26]Zhang RC,Li W,Pan SJ,et al.Application of chemical mutagenesis in improving germplasm resource.Mol Plant Breed,2017,15(12):5189-5196(in Chinese).张瑞成,李魏,潘素君,等.化学诱变在种质资源改良上的应用.分子植物育种,2017,15(12):5189-5196.
[27]Shan CJ,Liang ZS.Study on water physiological characteristics of black locust seedling under soil drought condition.J Shandong Agric Univ:Nat Sci Ed,2006,37(4):598-602(in Chinese).单长卷,梁宗锁.土壤干旱对刺槐幼苗水分生理特征的影响.山东农业大学学报:自然科学版,2006,37(4):598-602.
[28]Yang YH,Ma X,Zhang ZS,et al.Effects of drought stress on photosynthetic characteristics of wheat near-isogenic lines with different wax contents.Sci Agric Sin,2018,51(22):4241-4251(in Chinese).杨彦会,马晓,张子山,等.干旱胁迫对蜡质含量不同小麦近等基因系光合特性的影响.中国农业科学,2018,51(22):4241-4251.
[2]Chen MN,Chi XY,Pan LJ,et al.The development progress and prospects of peanut breeding in China.Chin Agric Sci Bull,2014,30(9):1-6(in Chinese).陈明娜,迟晓元,潘丽娟,等.中国花生育种的发展历程与展望.中国农学通报,2014,30(9):1-6.
[3]Chen SL.Identification and functional analysis of lipid biosynthesis related genes in peanut(Arachis hypogaea L.)[D].Beijing:Chinese Academy of Agricultural Sciences,2012(in Chinese).陈四龙.花生油脂合成相关基因的鉴定与功能研究[D].北京:中国农业科学院,2012.
[4]Yu SL.Genetics and Breeding of Peanuts in China.Shanghai:Shanghai Science and Technology Press,2011(in Chinese).禹山林.中国花生遗传育种学.上海:上海科学技术出版社,2011.
[5]Wang XJ.Studies on Peanut Biotechnology.Beijing:Science Press,2015(in Chinese).王兴军.花生生物技术研究.北京:科学出版社,2015.
[6]Yu SL.Varieties and Their Pedigree of Peanuts in China.Shanghai:Shanghai Science and Technology Press,2008(in Chinese).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.
[7]Ao Y,Pan QH.Phenotypic variation of Euonymus fortunei branches treated by 60Coγ-rays irradiation.JNucl Agric Sci,2008,22(3):271-275(in Chinese).敖妍,潘青华.60Coγ射线辐照扶芳藤枝条的表型变异研究.核农学报,2008,22(3):271-275.
[8]Yu XL,Liu LX,Qiao LX,et al.Effects of mixed high energy particle field on embryonic leaflet culture and plant regeneration of peanut.J Nucl Agric Sci,2012,26(3):433-438(in Chinese).于新玲,刘录祥,乔利仙,等.高能混合粒子场辐照对花生胚小叶组织培养及植株再生的影响.核农学报,2012,26(3):433-438.
[9]Wang JS,Sui JM,Xie YD,et al.Generation of peanut mutants by fast neutron irradiation combined with in vitro culture.J Radiat Res,2015,56(3):437-445.
[10]Wang JS,Zhao MX,Qiao LX,et al.Effects of fast neutron irradiation on plant regeneration in embryonic leaflet culture of peanut(Arachis hypogaea L.).Chin JOil Crop Sci,2013,35(2):148-152(in Chinese).王晶珊,赵明霞,乔利仙,等.快中子辐照对花生种子胚小叶植株再生的影响.中国油料作物学报,2013,35(2):148-152.
[11]Wang JS,Qiao LX,Zhao LS,et al.Performance of peanut mutants and their offspring generated from mixed high-energy particle field radiation and tissue culture.Genet Mol Res,2015,14(3):10837-10848.
[12]Wang Y,Qiao LX,Wu XL,et al.Effect of in vitro mutagenesis with pingyangmycin and NaCl-directed screening on production and quality of offspring in peanut.Acta Agric Boreali-Sin,2015,30(1):202-206(in Chinese).王亚,乔利仙,武秀玲,等.平阳霉素诱变与NaCl定向筛选对花生后代产量和品质性状的影响.华北农学报,2015,30(1):202-206.
[13]Rai MK,Kalia RK,Singh R,et al.Developing stress tolerant plants through in vitro selection-an overview of the recent progress.Environ Exp Bot,2011,71(1):89-98.
[14]Wu WG,Liu GR,Yang XJ.Applications of the mutation in connection with in vitro culture for plant breeding.Chin Agric Sci Bull,2005,21(11):197-201(in Chinese).吴伟刚,刘桂茹,杨学举.诱变与组织培养相结合在植物育种中的应用.中国农学通报,2005,21(11):197-201.
[15]Patade VY,Suprasanna P.An in vitro radiation induced mutagenesis-selection system for salinity tolerance in sugarcane.Sugar Tech,2009,11(3):246-251.
[16]Luo J,Zhou HC,Wang YQ,et al.EMS mutagenesis in vitro and screening of calli from strawberry leaves of resistance to Botrytis chinerea Pers.J Nucl Agric Sci,2009,23(1):90-94(in Chinese).罗静,周厚成,王永清,等.EMS离体诱变及抗草莓灰霉病愈伤组织的筛选.核农学报,2009,23(1):90-94.
[17]Chen L,Dong JW,Tang Y,et al.Selection of salt-tolerant mutants from ethyl methanesulfonatemutagenized poplar embryonic calli.Acta Agric Shanghai,2007,23(3):86-91(in Chinese).陈丽,董举文,唐寅,等.EMS诱变处理定向筛选杨树耐盐突变体研究.上海农业学报,2007,23(3):86-91.
[18]Li H,Li B,Zhao HB,et al.Study on alkalinity resistance of alfalfa callus in mutagenic treatments.Pratac Sci,2009,26(7):32-35(in Chinese).李红,李波,赵洪波,等.诱变处理苜蓿愈伤组织抗碱性的研究.草业科学,2009,26(7):32-35.
[19]Zhao MX,Sun HY,Ji RR,et al.In vitro mutagenesis and directed screening for salt-tolerant mutants in peanut.Euphytica,2013,193(1):89-99.
[20]Wang X,Li Y,Zhu H,et al.Directional screening on peanut mutant with high oil content and correlation between oil content and leaf water potential.J Qingdao Agric Univ,2019,36(1):30-33(in Chinese).王霞,李艳,朱虹,等.花生高油突变体的定向筛选及籽仁含油率与叶片水势的相关性分析.青岛农业大学学报,2019,36(1):30-33.
[21]Zhang DD,Wang WH,Li G,et al.Chemistry mutagenesis in vitro and characteristics of mutants in sweet potato.Plant Physiol J,2016,52(3):343-348(in Chinese).张丹丹,王维华,李冠,等.甘薯化学离体诱变及突变体的性状表现.植物生理学报,2016,52(3):343-348.
[22]Yan ZM,Feng YN,Han YL,et al.Effects of exogenous proline on proline metabolism of Cucumis melo under salt stress.Acta Bot Bor Occid Sin,2015,35(10):2035-2041(in Chinese).颜志明,冯英娜,韩艳丽,等.外源脯氨酸对盐胁迫下甜瓜脯氨酸代谢的影响.西北植物学报,2015,35(10):2035-2041.
[23]Gillman JD,Tetlow A,Hagely K,et al.Identification of the molecular genetic basis of the low palmitic acid seed oil trait in soybean mutant line rg3 and association analysis of molecular markers with elevated seed stearic acid and reduced seed palmitic acid.Mol Breed,2014,34(2):447-455.
[24]Zhang YB,Xiao L,Dong C,et al.Comparative study on mutagenic effects of pingyangmycin and NaN3 on wheat.J Henan Instit Sci Technol:Nat Sci Ed,2015,43(5):6-9(in Chinese).张彦波,肖磊,董策,等.平阳霉素和NaN3对小麦诱变效应的比较研究.河南科技学院学报:自然科学版,2015,43(5):6-9.
[25]Zeng XH.Comparing effectiveness of different mutagens for seed quality and analysis of mutants in Brassica napus[D].Wuhan:Huazhong Agricultural University,2010(in Chinese).曾新华.不同诱变方法对油菜种子诱变效果及突变体的研究[D].武汉:华中农业大学,2010.
[26]Zhang RC,Li W,Pan SJ,et al.Application of chemical mutagenesis in improving germplasm resource.Mol Plant Breed,2017,15(12):5189-5196(in Chinese).张瑞成,李魏,潘素君,等.化学诱变在种质资源改良上的应用.分子植物育种,2017,15(12):5189-5196.
[27]Shan CJ,Liang ZS.Study on water physiological characteristics of black locust seedling under soil drought condition.J Shandong Agric Univ:Nat Sci Ed,2006,37(4):598-602(in Chinese).单长卷,梁宗锁.土壤干旱对刺槐幼苗水分生理特征的影响.山东农业大学学报:自然科学版,2006,37(4):598-602.
[28]Yang YH,Ma X,Zhang ZS,et al.Effects of drought stress on photosynthetic characteristics of wheat near-isogenic lines with different wax contents.Sci Agric Sin,2018,51(22):4241-4251(in Chinese).杨彦会,马晓,张子山,等.干旱胁迫对蜡质含量不同小麦近等基因系光合特性的影响.中国农业科学,2018,51(22):4241-4251.