三角梅扦插枝化学诱变育种
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摘要
三角梅是一种重要的热带和亚热带观赏植物。由于三角梅有性繁殖困难,诱变育种是目前比较可行的育种方式。本研究采用甲基磺酸乙酯(EMS)、叠氮化钠(NaN3)和平阳霉素(PYM)3种不同类型诱变剂对‘伊娃夫人’和‘马尼拉小姐’2个三角梅品种扦插枝进行了化学诱变。研究发现‘伊娃夫人’和‘马尼拉小姐’20 h EMS半致死剂量分别为0.22%和0.62%。随EMS浓度增加,2个品种扦插枝发芽数和最长分枝长呈下降趋势,叶绿素和类胡萝卜素含量逐渐增加;‘伊娃夫人’叶片逐渐向暗色调变化,‘马尼拉小姐’叶片向红色色调变化。20 h酸性NaN3处理下,‘伊娃夫人’无法存活,‘马尼拉小姐’成活率为82%～92%;随NaN3浓度升高,‘马尼拉小姐’发芽数呈显著下降趋势。相比对照,20 h PYM显著提高了2个三角梅品种扦插枝最大分枝长度,但对扦插枝存活率无显著影响。中高浓度EMS和PYM处理‘马尼拉小姐’三角梅出现了叶缘缺刻、叶表褶皱和双叶尖等变异株。
Bougainvillea Comm. ex Juss is an important ornamental plant in the tropical and sub-tropical regions of China. Because the sexual reproduction of Bougainvillea is difficult, mutation breeding is an available breeding method.The present study used ethylmethane sulfonate(EMS), sodium azide(NaN3) and pingyangmycin(PYM) to mutagenize the cuttings of B. glabra 'Mrs. Eva' and B. × buttiana 'Miss Manila', respectively. In 20 h EMS treatments, the half lethal dose of 'Mrs. Eva' and 'Miss Manila' cuttings was 0.22% and 0.62%, respectively. With the increasing of EMS concentration, the shoot number and length of the longest branch of two Bougainvillea varieties showed a decreasing trend, while the chlorophyll and carotenoid content increased gradually. With the increasing of EMS concentration, the leaf hue of 'Mrs. Eva' changed to grayish tone and that of 'Miss Manila' changed to red tone gradually. Almost all 'Mrs.Eva' cuttings died in 20 h NaN3 treatments, while the survival rate of 'Miss Manila' cuttings was 82%-92%. With the increasing of NaN3 concentration, the shoot number of 'Miss Manila' decreased significantly. 20 h PYM treatments significantly improved the length of the longest branch of two Bougainvillea varieties in contrast to the control, but had no significant influence on the survival rate. Moreover, variant seedlings were found in EMS and PYM treatments, representing transformative leaf margin, wrinkled leaf surface or double leaf tips.
Bougainvillea Comm. ex Juss is an important ornamental plant in the tropical and sub-tropical regions of China. Because the sexual reproduction of Bougainvillea is difficult, mutation breeding is an available breeding method.The present study used ethylmethane sulfonate(EMS), sodium azide(NaN3) and pingyangmycin(PYM) to mutagenize the cuttings of B. glabra 'Mrs. Eva' and B. × buttiana 'Miss Manila', respectively. In 20 h EMS treatments, the half lethal dose of 'Mrs. Eva' and 'Miss Manila' cuttings was 0.22% and 0.62%, respectively. With the increasing of EMS concentration, the shoot number and length of the longest branch of two Bougainvillea varieties showed a decreasing trend, while the chlorophyll and carotenoid content increased gradually. With the increasing of EMS concentration, the leaf hue of 'Mrs. Eva' changed to grayish tone and that of 'Miss Manila' changed to red tone gradually. Almost all 'Mrs.Eva' cuttings died in 20 h NaN3 treatments, while the survival rate of 'Miss Manila' cuttings was 82%-92%. With the increasing of NaN3 concentration, the shoot number of 'Miss Manila' decreased significantly. 20 h PYM treatments significantly improved the length of the longest branch of two Bougainvillea varieties in contrast to the control, but had no significant influence on the survival rate. Moreover, variant seedlings were found in EMS and PYM treatments, representing transformative leaf margin, wrinkled leaf surface or double leaf tips.
引文
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[8]张蒙蒙.枇杷潜伏芽的EMS诱变及ISSR分析[D].成都:四川农业大学,2013.
[9]罗丽.利用EMS诱变获得甜橙抗溃疡病突变体的研究[D].长沙:湖南农业大学,2015.
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[12]陈宇.平阳霉素诱导尾巨桉突变及突变体耐寒性的初步探讨[J].亚热带农业研究,2013,9(3):173-176.
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[14]Cai Y,Sun M,Wu H,et al.Characterization and quantification of betacyanin pigments from diverse Amaranthus species[J].Journal of Agricultural&Food Chemistry,1998,46(6):2063-2070.
[15]周玉丽.甜叶菊EMS诱变及耐盐突变体的筛选与鉴定[D].合肥:安徽农业大学,2013.
[16]任磊,赵夏陆,许靖,等.4种茶菊对干旱胁迫的形态和生理响应[J].生态学报,2015,35(15):5131-5139.
[17]Fina J,Casadevall R,Abdelgawad H,et al.UV-B inhibits leaf growth through changes in growth regulating factors and gibberellin levels[J].Plant Physiology,2017,174(2):1110-1126.
[18]刘彧.自然生境下不同表型盐地碱蓬色素积累及光合特性的研究[D].济南:山东师范大学,2006.
[19]唐龙翔,康少杰,李文庆.干旱胁迫对果树开花的诱导及对生理生化性状的影响[J].北方园艺,2009,9:111-115.
[20]Blein T,Pautot V,Laufs P.Combinations of mutations sufficient to alter Arabidopsis leaf dissection[J].Plants,2013,2(2):230-247.
[21]Serrano-Cartagena J,Candela H,Robles P,et al.Genetic analysis of incurvata mutants reveals three independent genetic operations at work in Arabidopsis leaf morphogenesis[J].Genetics,2000,156(3):1363.
[22]Pinon V,Etchells J P,Rossignol P,et al.Three PIGGYBACK genes that specifically influence leafpatterning encode ribosomal proteins[J].Development,2008,135(7):1315-1324.
[23]狄佳春,赵亮,陈旭升.棉花叶片突变体的研究进展[J].中国农学通报,2016,32(36):78-86.
[24]Rinaldi M A,Liu J,Enders T A,et al.A gain-of-function mutation in IAA16 confers reduced responses to auxin and abscisic acid and impedes plant growth and fertility[J].Plant Molecular Biology,2012,79(4-5):359-373.
[25]Wang D,Yuan S,Yin L,et al.A missense mutation in the transmembrane domain of CESA9 affects cell wall biosynthesis and plant growth in rice[J].Plant Science,2012,196(11):117-124.
[26]Chen H,Cheng Z,Ma X,et al.A knockdown mutation of YELLOW-GREEN LEAF2 blocks chlorophyll biosynthesis in rice[J].Plant Cell Reports,2013,32(12):1855-1867.
[27]Suzuki M,Miyahara T,Tokumoto H,et al.Transposon-mediated mutation of CYP76AD3 affects betalain synthesis and produces variegated flowers in four o'clock(Mirabilis jalapa)[J].Journal of Plant Physiology,2014,171(17):1586-1590.
[28]Gupta M N.Mutation breeding in Bougainvillea[J].Indian Journal of Genetics Plant Breeding,1974,34:1295-1299.
[29]陈庭,范雅文,刘伟.Co60-γ射线诱变宝巾(Bougainvillea)的生物学效应研究[J].河南农业科学,2012,41(6):133-136.
[30]孙利娜,王华新,龚建英,等.宝巾花辐射诱变效应研究初报[J].北方园艺,2013,19:91-93.
[31]Swaroop K,Jain R,Janakiram T.Effect of different doses of gamma rays for induction of mutation in Bougainvillea cv Mahatma Gandhi[J].Indian Journal of Agricultural Sciences,2015,85(9):1245-1247.
[32]Roy R,Singh S,Rastogi R.Bougainvillea:Identification,gardening and landscape use[M].Lucknow:Army Printing Press,2015.
[33]Shu Q Y,Forster B P,Nakagawa H.Plant Mutation Breeding&Biotechnology[M].Vienna:FAO/IAEA Division of Nuclear Techniques in Food and Agriculture,2012:169-180.
[2]张瑞成,李魏,潘素君,等.化学诱变在种质资源改良上的应用[J].分子植物育种,2017,15(10):5189-5196.
[3]刘翔.EMS诱变技术在植物育种中的研究进展[J].激光生物学报.2014,23:197-201.
[4]Khan S,Al-Qurainy F,Anwar F.Sodium Azide:a chemical mutagen for enhancement of agronomic traits of crop plants[J].Environ.We Int.J.Sci.Tech.,2009,4:1-21.
[5]韩永亮,李世云,路正营,等.平阳霉素在作物育种中的应用[J].农技服务,2013,30(8),900.
[6]陈庭,王爱敏,刘运权,等.NaN3对宝巾的诱变效应初步研究[J].中国农学通报,2012,28(25):191-195.
[7]范芳绮.九重葛之组织培养化学诱变[D].台中:中兴大学,2012.
[8]张蒙蒙.枇杷潜伏芽的EMS诱变及ISSR分析[D].成都:四川农业大学,2013.
[9]罗丽.利用EMS诱变获得甜橙抗溃疡病突变体的研究[D].长沙:湖南农业大学,2015.
[10]祁伟.红掌离体化学诱变技术的研究[D].苏州:苏州大学,2016.
[11]王红,陈发棣,赵宏波,等.平阳霉素对盆栽小菊意大利红的诱变效应[J].南京农业大学学报,2007,30(1):39-43.
[12]陈宇.平阳霉素诱导尾巨桉突变及突变体耐寒性的初步探讨[J].亚热带农业研究,2013,9(3):173-176.
[13]Lichtenthaler H K.Chlorophylls and carotenoids:Pigments of photosynthetic biomembranes[J].Methods in Enzymology,1987,148:350-383.
[14]Cai Y,Sun M,Wu H,et al.Characterization and quantification of betacyanin pigments from diverse Amaranthus species[J].Journal of Agricultural&Food Chemistry,1998,46(6):2063-2070.
[15]周玉丽.甜叶菊EMS诱变及耐盐突变体的筛选与鉴定[D].合肥:安徽农业大学,2013.
[16]任磊,赵夏陆,许靖,等.4种茶菊对干旱胁迫的形态和生理响应[J].生态学报,2015,35(15):5131-5139.
[17]Fina J,Casadevall R,Abdelgawad H,et al.UV-B inhibits leaf growth through changes in growth regulating factors and gibberellin levels[J].Plant Physiology,2017,174(2):1110-1126.
[18]刘彧.自然生境下不同表型盐地碱蓬色素积累及光合特性的研究[D].济南:山东师范大学,2006.
[19]唐龙翔,康少杰,李文庆.干旱胁迫对果树开花的诱导及对生理生化性状的影响[J].北方园艺,2009,9:111-115.
[20]Blein T,Pautot V,Laufs P.Combinations of mutations sufficient to alter Arabidopsis leaf dissection[J].Plants,2013,2(2):230-247.
[21]Serrano-Cartagena J,Candela H,Robles P,et al.Genetic analysis of incurvata mutants reveals three independent genetic operations at work in Arabidopsis leaf morphogenesis[J].Genetics,2000,156(3):1363.
[22]Pinon V,Etchells J P,Rossignol P,et al.Three PIGGYBACK genes that specifically influence leafpatterning encode ribosomal proteins[J].Development,2008,135(7):1315-1324.
[23]狄佳春,赵亮,陈旭升.棉花叶片突变体的研究进展[J].中国农学通报,2016,32(36):78-86.
[24]Rinaldi M A,Liu J,Enders T A,et al.A gain-of-function mutation in IAA16 confers reduced responses to auxin and abscisic acid and impedes plant growth and fertility[J].Plant Molecular Biology,2012,79(4-5):359-373.
[25]Wang D,Yuan S,Yin L,et al.A missense mutation in the transmembrane domain of CESA9 affects cell wall biosynthesis and plant growth in rice[J].Plant Science,2012,196(11):117-124.
[26]Chen H,Cheng Z,Ma X,et al.A knockdown mutation of YELLOW-GREEN LEAF2 blocks chlorophyll biosynthesis in rice[J].Plant Cell Reports,2013,32(12):1855-1867.
[27]Suzuki M,Miyahara T,Tokumoto H,et al.Transposon-mediated mutation of CYP76AD3 affects betalain synthesis and produces variegated flowers in four o'clock(Mirabilis jalapa)[J].Journal of Plant Physiology,2014,171(17):1586-1590.
[28]Gupta M N.Mutation breeding in Bougainvillea[J].Indian Journal of Genetics Plant Breeding,1974,34:1295-1299.
[29]陈庭,范雅文,刘伟.Co60-γ射线诱变宝巾(Bougainvillea)的生物学效应研究[J].河南农业科学,2012,41(6):133-136.
[30]孙利娜,王华新,龚建英,等.宝巾花辐射诱变效应研究初报[J].北方园艺,2013,19:91-93.
[31]Swaroop K,Jain R,Janakiram T.Effect of different doses of gamma rays for induction of mutation in Bougainvillea cv Mahatma Gandhi[J].Indian Journal of Agricultural Sciences,2015,85(9):1245-1247.
[32]Roy R,Singh S,Rastogi R.Bougainvillea:Identification,gardening and landscape use[M].Lucknow:Army Printing Press,2015.
[33]Shu Q Y,Forster B P,Nakagawa H.Plant Mutation Breeding&Biotechnology[M].Vienna:FAO/IAEA Division of Nuclear Techniques in Food and Agriculture,2012:169-180.