棉花光敏雄性核不育突变体PSM4创制及其特性研究
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摘要
为了创造棉花光敏雄性不育系材料,解决杂交制种中人工去雄成本高的问题,自2006年起进行了大规模组织培养,创造棉花突变体,以期筛选出棉花光敏雄性不育材料。2012年在组织培养再生植株后代中发现新型光敏核不育材料PSM4(photoperiod sensitive male sterility mutant of cotton),并于2012~2017年在海南三亚吉阳镇中寥村中棉所南繁基地和安阳中棉所试验田进行材料选育、生长发育调查及光周期实验。结果表明:(1)PSM4在日照时数大于12h时表现为雄性不育;在日照时数为11.5~12h时表现为有少量花粉,处于育性转变期;在日照时数小于11.5h时表现为正常可育。(2)遮光试验显示,PSM4花药败育关键时期为开花前12~15d;石蜡切片显微观察显示,花药败育原因是在长日照条件下花粉壁外层物质缺失引起的。(3)遗传规律研究显示,PSM4育性受单隐性基因控制,不受阴雨气候条件影响。(4)用6个棉花品系与PSM4进行正反交试验发现F1代均可育,F2代分离出光敏核不育单株,分离比符合3∶1的单隐性分离规律。研究认为,新型棉花光敏核不育材料PSM4农艺性状优良,其光敏不育突变为隐性性状,所有的陆地棉品种(系)均为其恢复系,有利于杂交种的培育和棉花混选混交育种群体的建立,具有广阔的应用前景。
To solve the problem of high cost for traditional method of hybrid seed production which involves hand emasculation followed by pollination,a large-scale cotton tissue culture was carried out to creat a cotton mutant and developed a highly sensitive genetic male sterility since 2006.In 2012,aphotoperiod sensitive genetic male sterility was screened out in their offspring which named PSM4(Photoperiod sensitive genetic male sterility mutant of cotton).During 2012-2017,breeding,material selection,the growth and development investigation and characteristics of photoperiod were carry out in the experimental field of CRI(Cotton Research Institution of China)in Anyang,He'nan Province and South Crop Breeding Base in Zhongliao Village,Jiyang Town,Sanya,Hainan Province.(1)PSM4 shows male sterility when the sun-shine duration is more than 12 h;when between 11.5-12 h,it is in the fertility transformation stage;when less than 11.5 h,it shows normal fertility.(2)Shading experiments show that the crucialtime of anther and pollen abortion at 12-15 days before anthesis.(3)Paraffin section of anther shows the reason of pollen abortion is the lost of extine under long daylight exposure.(4)All individuals were fertile in F1 population from the reciprocal crosses between PSM4 and six upland lines and the results shows that the photoperiod sensitive genetic male sterility character is controlled by their nuclear genes.Genetic studies indicate that PSM4 fertility was controlled by single recessive genes for the segregation of fertile and sterile plants ratio of 3∶1 in F2 population and unaffected by the rainy weather.The successful cultivation of PSM4 will greatly advance the study of hybrid seed production and use in an industry revolution in cotton breeding.
To solve the problem of high cost for traditional method of hybrid seed production which involves hand emasculation followed by pollination,a large-scale cotton tissue culture was carried out to creat a cotton mutant and developed a highly sensitive genetic male sterility since 2006.In 2012,aphotoperiod sensitive genetic male sterility was screened out in their offspring which named PSM4(Photoperiod sensitive genetic male sterility mutant of cotton).During 2012-2017,breeding,material selection,the growth and development investigation and characteristics of photoperiod were carry out in the experimental field of CRI(Cotton Research Institution of China)in Anyang,He'nan Province and South Crop Breeding Base in Zhongliao Village,Jiyang Town,Sanya,Hainan Province.(1)PSM4 shows male sterility when the sun-shine duration is more than 12 h;when between 11.5-12 h,it is in the fertility transformation stage;when less than 11.5 h,it shows normal fertility.(2)Shading experiments show that the crucialtime of anther and pollen abortion at 12-15 days before anthesis.(3)Paraffin section of anther shows the reason of pollen abortion is the lost of extine under long daylight exposure.(4)All individuals were fertile in F1 population from the reciprocal crosses between PSM4 and six upland lines and the results shows that the photoperiod sensitive genetic male sterility character is controlled by their nuclear genes.Genetic studies indicate that PSM4 fertility was controlled by single recessive genes for the segregation of fertile and sterile plants ratio of 3∶1 in F2 population and unaffected by the rainy weather.The successful cultivation of PSM4 will greatly advance the study of hybrid seed production and use in an industry revolution in cotton breeding.
引文
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[21]刘子涵,石晓艺,闫鹏娇,等.D~2型细胞质雄性不育小麦绒毡层细胞程序化死亡与活性氧代谢[J].中国农业科学,2017,50(21):4 071-4 086.LIU Z H,SHI X Y,YAN P J,et al.Tapetal programmed cell death,antioxidant response and oxidative stress in wheat anthers associated with D2-type cytoplasmic male-sterility[J].Scientia Agricultura Sinica,2017,50(21):4 071-4 086.
[22]张姣,朱启迪,巨岚,等.小麦生理型雄性不育系微丝骨架和胼胝质的变化与其相关基因的表达分析[J].中国农业科学,2015,48(14):2 687-2 696.ZHANG J,ZHU Q D,JU L,et al.Organization changes of actin cytoskeleton and callose with the expression analysis of related genes in physiological male sterile wheat[J].Scientia Agricultura Sinica,2015,48(14):2 687-2 696.
[23]张保雷,张卫东,高庆荣,等.温光敏雄性不育小麦BNS育性的遗传效应分析[J].中国农业科学,2013,46(8):1 533-1 542.ZHANG B L,ZHANG W D,GAO Q R,et al.Genetic analysis on male sterility of thermo-photo-sensitive male sterile line BNS in wheat[J].Scientia Agricultura Sinica,2013,46(8):1 533-1 542.
[24]CHEN X D,SUN D F,RONG D F,et al.A recessive gene controlling male sterility sensitive to short daylength/low temperature in wheat(Triticum aestivum L.)[J].J.Zhejiang Univ.Sc.B,2011,12(11):943-950.
[25]MA J H,WEI H L,LIU J,et al.Selection and characterization of a novel photoperiod-sensitive male sterile line in upland cotton[J].Journal of Integrative Plant Biology,2013,55(7):608-618.
[26]刘金兰,聂以春,黄观武,等.棉花洞A型核雄性不育材料花粉发育的细胞形态学观察[J].棉花学报,1994,6(2):70-73.LIU J L,NIE Y C,HUANG G W,et al.A cytological study on pollen development of nuclear male sterile line“Dong A”of cotton[J].Acta Gossypii Sinica,1994,6(2):70-73.
[27]吕川根,邹江石.两系法杂交稻两优培九育种的理论与实践[J].中国农业科学,2016,49(9):1 635-1 645.LC G,ZOU J S.Theory and practice on breeding of twoline hybrid rice,Liangyoupeijiu[J].Scientia Agricultura Sinica,2016,49(9):1 635-1 645.
[28]张华丽,陈晓阳,黄建中,等.中国两系杂交水稻光温敏核不育基因的鉴定与演化分析[J].中国农业科学,2015,48(1):1-9.ZHANG H L,CHEN X Y,HUANG J Z,et al.Identification and transition analysis of photo-/thermo-sensitive genic male sterile genes in two-line hybrid rice in China[J].Scientia Agricultura Sinica,2015,48(1):1-9.
[29]SINGH VK,UPADHYAY P,SINHA P,et al.Determination of genetic relationships among elite thermosensitive genic male sterile lines(TGMS)of rice(Oryza sativa L.)employing morphological and simple sequence repeat(SSR)markers[J].J.Genet,2011,90(1):11-19.
[30]张朝军,范术丽,武芝霞,等.棉花大田植株叶柄组织培养体系的建立[J].西北植物学报,2011,31(6):1 257-1 263.ZHANG C J,FAN S L,WU Z X,et al.Plantlets regeneration from mature leafs petiole of cotton[J].Acta Botanica Boreali-Occidentalia Sinica,2011,31(6):1 257-1 263.
[31]周志林,聂以春,张献龙,等.棉花体细胞培养中染色体的变异[J].江苏农业学报,2008,24(2):126-129.ZHO Z L,NIE Y C,ZHANG X L,et al.Chromosomal variations during culture of somatic cells of cotton[J].Jiangsu Journal of Agricultural Sciences,2008,24(2):126-129.
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[2]叶祯维,邓晓英,石玉真,等.杂交棉中棉所70后代分离群体产量品质的表型变异分析[J].棉花学报,2016,28(1):1-10.YE W Z,DENG X Y,SHI Y Z,et al.Analysis of phenotypic variation in fiber yield and quality in segregative populations of hybrid cotton CCRI 70[J].Cotton Science,2016,28(1):1-10.
[3]GALAU G A,WILKINS T A.Alloplasmic male sterility in AD allotetraploid Gossypium hirsutum upon replacement of its resident A cytoplasm with that of D species G.harknessii[J].TAG,1989,78(1):23-30.
[4]马勇,吴建勇,邢朝柱,等.哈克尼西棉细胞质雄性不育系和保持系差异表达基因分析[J].中国农业科学,2009,42(10):3 706-3 712.MA Y,WU J Y,XING C Z,et al.Differentially expressed genes between Cytoplasmic male sterility lines and maintainer lines of Gossypium harknessii[J],Scintia agricultura sinica,2009,42(10):3 706-3 712.
[5]MEYER V G,MEYER J R.Cytoplasmically controlled male sterility in cotton[J].Crop Science,1965,5(5):444-448.
[6]MEVER V G.Some effects of genes,cytoplasm,and environment on male sterility of cotton(Gossypium)[J].Crop Science,1969,9(2):237-242.
[7]MEYER V G.Development of B-lines for two cytoplasmically controlled male sterilities of cotton[J].Crop Science,1970,10(6):720-721.
[8]MEYER V G.Registration of sixteen germplasm lines of upland cotton(Reg.No.GP 3to GP 18)[J].Crop Science,1973,13(6):778.
[9]MEYER V G.Male sterility from Gossypium harknessii[J].Journal of Heredity,1975,66(1):23-27.
[10]STEWART J M.A new cytoplasmic male sterile and restorer for cotton[J].Beltwide Cotton Production;1992:610.
[11]WU J Y,CAO X X,GUO L P,et al.Development of a candidate gene marker for Rf(1)based on a PPR gene in cytoplasmic male sterile CMS-D2 Upland cotton[J].Molecular Breeding,2014,34(1):231-240.
[12]WANG F,STEWART J M,ZHANG J.Molecular markers linked to the Rf(2)fertility restorer gene in cotton[J].Genome,2007,50(9):818-824.
[13]豆丽萍,王庆亚,唐灿明.陆地棉双隐性核雄性不育系ms5ms6花药发育过程的研究[J].棉花学报,2009,21(4):265-270.DOU L P,WANG Q Y,TANG C M.The anther development research on double recessiveness nuclear male-sterile Line ms5ms6of Gossypium hirsutum L.[J].Acta Gossypii Sinica,2009,21(4):265-270.
[14]张天真,冯义军,潘家驹.我国发现的4个棉花核雄性不育系的遗传分析[J].棉花学报,1992,4(1):1-8.ZHANG T Z,FENG Y J,PAN J J.Genetic analysis of four gentic male sterile lines of cotton discovered in China[J].Acta Gossypii Sinica,1992,4(1):1-8.
[15]CHEN D,DING Y,GUO W,et al.Molecular mapping of genic male-sterile genes ms(15),ms(5)and ms(6)in tetraploid cotton[J].Plant Breeding,2009,128(2):193-198.
[16]SEKHAR L,KHADI B M,PATIL R S,et al.Biochemical and molecular dissection of thermo-sensitive genetic male sterility in diploid cotton(Gossypiuni arboreum L.)[J].Journal of Environmental Biology,2016,37(4):579-583.
[17]YUE J J,REN Y,WU S J,et al.Differential proteomic studies of the genic male-sterile line and fertile line anthers of upland cotton(Gossypium hirsutum L.)[J].Genes Genom,2014,36(4):415-426.
[18]周仲华,陈金湘,刘爱玉,等.棉花温敏雄性不育系特棉S-1的形态学和细胞学特征研究[J].棉花学报,2008,20(6):408-413.ZHOU Z H,CHEN J X,LIU A Y,et al.Studies on morphological and cytological features of thermo-sensitive sterile line temian S-1(Gossypium hirsutum)[J].Cotton Science,2008,20(6):408-413.
[19]邵圣才,文彬,顾相蕊.棉花光敏雄性核不育两用系研究与利用[J].棉花科学,2012,34(1):14-16.SHAO S C,WEN B,GU X R.Research and utilization to cotton photoperiod sensitive male sterile[J].Cotton Sciences,2012,34(1):14-16.
[20]何觉民,戴君惕,邹应斌,等.生态遗传雄性不育理论与两系杂交植物Ⅰ.生态遗传雄性不育理论[J].作物杂志,1994,(1):17-19.HE J M,DAI J T,ZOU Y B,et al.The theory of ecological-genetical sterility and hybrid crop produced two-line method[J].Crops,1994,(1):17-19.
[21]刘子涵,石晓艺,闫鹏娇,等.D~2型细胞质雄性不育小麦绒毡层细胞程序化死亡与活性氧代谢[J].中国农业科学,2017,50(21):4 071-4 086.LIU Z H,SHI X Y,YAN P J,et al.Tapetal programmed cell death,antioxidant response and oxidative stress in wheat anthers associated with D2-type cytoplasmic male-sterility[J].Scientia Agricultura Sinica,2017,50(21):4 071-4 086.
[22]张姣,朱启迪,巨岚,等.小麦生理型雄性不育系微丝骨架和胼胝质的变化与其相关基因的表达分析[J].中国农业科学,2015,48(14):2 687-2 696.ZHANG J,ZHU Q D,JU L,et al.Organization changes of actin cytoskeleton and callose with the expression analysis of related genes in physiological male sterile wheat[J].Scientia Agricultura Sinica,2015,48(14):2 687-2 696.
[23]张保雷,张卫东,高庆荣,等.温光敏雄性不育小麦BNS育性的遗传效应分析[J].中国农业科学,2013,46(8):1 533-1 542.ZHANG B L,ZHANG W D,GAO Q R,et al.Genetic analysis on male sterility of thermo-photo-sensitive male sterile line BNS in wheat[J].Scientia Agricultura Sinica,2013,46(8):1 533-1 542.
[24]CHEN X D,SUN D F,RONG D F,et al.A recessive gene controlling male sterility sensitive to short daylength/low temperature in wheat(Triticum aestivum L.)[J].J.Zhejiang Univ.Sc.B,2011,12(11):943-950.
[25]MA J H,WEI H L,LIU J,et al.Selection and characterization of a novel photoperiod-sensitive male sterile line in upland cotton[J].Journal of Integrative Plant Biology,2013,55(7):608-618.
[26]刘金兰,聂以春,黄观武,等.棉花洞A型核雄性不育材料花粉发育的细胞形态学观察[J].棉花学报,1994,6(2):70-73.LIU J L,NIE Y C,HUANG G W,et al.A cytological study on pollen development of nuclear male sterile line“Dong A”of cotton[J].Acta Gossypii Sinica,1994,6(2):70-73.
[27]吕川根,邹江石.两系法杂交稻两优培九育种的理论与实践[J].中国农业科学,2016,49(9):1 635-1 645.LC G,ZOU J S.Theory and practice on breeding of twoline hybrid rice,Liangyoupeijiu[J].Scientia Agricultura Sinica,2016,49(9):1 635-1 645.
[28]张华丽,陈晓阳,黄建中,等.中国两系杂交水稻光温敏核不育基因的鉴定与演化分析[J].中国农业科学,2015,48(1):1-9.ZHANG H L,CHEN X Y,HUANG J Z,et al.Identification and transition analysis of photo-/thermo-sensitive genic male sterile genes in two-line hybrid rice in China[J].Scientia Agricultura Sinica,2015,48(1):1-9.
[29]SINGH VK,UPADHYAY P,SINHA P,et al.Determination of genetic relationships among elite thermosensitive genic male sterile lines(TGMS)of rice(Oryza sativa L.)employing morphological and simple sequence repeat(SSR)markers[J].J.Genet,2011,90(1):11-19.
[30]张朝军,范术丽,武芝霞,等.棉花大田植株叶柄组织培养体系的建立[J].西北植物学报,2011,31(6):1 257-1 263.ZHANG C J,FAN S L,WU Z X,et al.Plantlets regeneration from mature leafs petiole of cotton[J].Acta Botanica Boreali-Occidentalia Sinica,2011,31(6):1 257-1 263.
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