长穗偃麦草幼穗离体培养高频再生体系的建立
|
摘要
以长穗偃麦草(Elytrigia elongata)幼穗为外植体, MS为基本培养基,研究了不同植物生长调节物质配比对其愈伤组织诱导、分化和生根的影响。结果表明,长穗偃麦草幼穗诱导最佳培养基为MC+3 mg·L-1 2,4-D,诱导率达66.67%, 4周后可见淡黄色愈伤组织。最佳分化培养基为MC+0.1 mg·L-1 2,4-D+3mg·L-1 6-BA,分化率为64.44%, 4周后出现芽点,同时伴随根毛发生。最佳生根培养基为MR+0.5 mg·L-1 NAA,生根率为100%,移栽后全部成活?
The tissue culture and rapid propagation system of Elytrigia elongata was established by using the panicle as explants and MS as the basic callus. The effects of various plant grouth regulators combinations on callus induction, differentiation and rooting were studied. The results showed that the optimal medium for induction of panicles of E. elongata was MC+3 mg·L-1 2,4-D, and the induction rate was 66.67%. After 4 weeks, light yellow callus was observed. The optimal differentiation medium was MC+0.1 mg·L-1 2,4-D+3 mg·L-16-BA, and the differentiation rate was 64.44%. After 4 weeks, the bud point appeared, accompanied by root hair. The best rooting medium was MR+0.5 mg·L-1 NAA, the rooting rate was 100%, and all survived after transplanting.
The tissue culture and rapid propagation system of Elytrigia elongata was established by using the panicle as explants and MS as the basic callus. The effects of various plant grouth regulators combinations on callus induction, differentiation and rooting were studied. The results showed that the optimal medium for induction of panicles of E. elongata was MC+3 mg·L-1 2,4-D, and the induction rate was 66.67%. After 4 weeks, light yellow callus was observed. The optimal differentiation medium was MC+0.1 mg·L-1 2,4-D+3 mg·L-16-BA, and the differentiation rate was 64.44%. After 4 weeks, the bud point appeared, accompanied by root hair. The best rooting medium was MR+0.5 mg·L-1 NAA, the rooting rate was 100%, and all survived after transplanting.
引文
Bao W,Mi FG(2007).A study on induction of emdryogenic callus and plant regeneration from mature embryos of Elytrigia hybrid(E.elongata×E.intermedia).Acta Agrect Sin,24(11):348-351(in Chinese with English abstract)[鲍芫,米福贵(2007).长穗偃麦草与中间偃麦草杂种成熟胚离体培养与再生体系的建立.草地学报,24(11):348-351]
Chen LP,Xue MF,Chen LZ,et al(2016).Preliminary study on regeneration plantlet from matured embryo of barnyardgrass by callus tissue culture.Plant Physiol J,52(12):1915-1920(in Chinese with English abstract)[陈丽萍,徐明飞,陈列忠等(2016).稗草成熟胚诱导愈伤组织再生植株的初步研究.植物生理学报,52(12):1915-1920]
Fan SH,Guo AG(2000).A study on the origin of HMW-GS 14 and 15 in Xiaoyan 6.J Northwest Agric Univ,28(6):1-5(in Chinese with English abstract)[范三红,郭蔼光(2000).小偃麦6号高分子质量麦谷蛋白14和15亚基来源分析.西北农业大学学报,28(6):1-5]
Gao SQ,Wang YB,Tang YM,et al(2011).Preliminary function research of EeNAC9 gene from Elytrigia elongata.Biotechnol Bull,(6):47-52(in Chinese with English abstract)[高世庆,王永波,唐益苗等(2011).长穗偃麦草EeNAC9基因功能初步研究.生物技术通报,(6):47-52]
Guo Q,Meng L,Mao PC,et al(2015).Salt tolerance in two tall wheatgrass species is associated with selective capacity for K+over Na+.Acta Physiol Plant,17081(37):1708-1760
Guo XY,Li HS,Peng KQ,et al(2011).Tissue culture and rapid propagation technology of Triathena lutarioriparia L.Liu sp.nov.Plant Physiol J,47(10):987-990(in Chinese with English abstract)[郭夏宇,李合松,彭克勤等(2011).南荻的组织培养与快速繁殖技术.植物生理学报,47(10):987-990]
He Y,Tian ZH,Zheng YL(2005).High frequency plant regeneration from mature embryos in vitro culture of tall fescue.Pratacult Sci,22(6):28-34(in Chinese with English abstract)[何勇,田志宏,郑用琏(2005).高羊茅成熟胚离体培养及高频植株再生.草业科学,22(6):28-34]
Huang K,Tang J,Liu ZZ,et al(2015).Influences of auxin and cytokine on propagation in vitro of Mulberry.J Southwest Univ,37(3):28-34(in Chinese with English abstract)[黄科,唐婧,刘自震等(2015).生长素和细胞分裂素对桑树离体繁育的影响.西南大学学报(自然科学版),37(3):28-34]
Huang W,Fu L,Li C,et al(2017).Quercetin,hyperin,and chlorogenicacid improve endothelial function by antioxidant,anti in flammatory,and ACE inhibitory effects.JFood Sci,82(5):1239-1246
Huo XW,Wei JH,Zhang H,et al(2004).Study of plant regeneration in wheatgrass(Agropyron Gaertn).Acta Agric Boreali Sin,19(1):17-20(in Chinese with English abstract)[霍秀文,魏建华,张辉等(2004).冰草属植物组织培养再生体系的建立.华北农学报,19(1):17-20]
Hussein Z,Dryanova A,Maret D,et al(2014).Gene expression analysis in the roots of salt-stressed wheat and the cytogenetic derivatives of wheat combined with the salt-tolerant wheatgrass,Lophopyrum elongatum.Plant Cell Rep,33(1):189-201
Jiang BN,Wang C,Pan JW(2014).Molecular regulatory mechanisms of hypocotyl elongation and phototropism in Arabidopsis.Plant Physiol J,50(10):1435-1444(in Chinese with English abstract)[姜楠,王超,潘建伟(1993).拟南芥下胚轴伸长与向光性的分子调控机理.植物生理学报,50(10):1435-1444]
Li WJ,Li XQ,Jia MM,et al(2012).Effects of 6-BA and 2,4-D on callus induction,growth and plantlet regeneration of shepherd’s purse[Capsella bursa-pastoris(L.)Medic].Plant Physiol J,48(2):141-146(in Chinese with English abstract)[李文静,李学强,贾毛毛等(2012).6-BA、NAA和2,4-D不同配比对荠菜愈伤组织诱导、生长及植株再生的影响.植物生理学报,48(2):141-146]
Li YJ,Liu JZ,Li B,et al(2010).Chromosomal location of the genes conferring the tolerance to phosphorus deficiency stress in Lophopyrum elongatum genome.J Genet Genomics,26(6):703-710(in Chinese with English abstract)[李玉京,刘建中,李滨等(2010).长穗偃麦草基因组中与耐低磷营养胁迫有关的基因的染色体定位.遗传学报,26(6):703-710]
Liu SY,Wang PW,Guan SY,et al(2013).Prelininary study on effects of 6-BA on callus induction of soybean.Hubei Agr Sci,52(16):3999-4001(in Chinese with English abstract)[刘思言,王丕武,关淑艳等(2013).6-BA对大豆愈伤组织诱导影响的初步研究.湖北农业科学,52(16):3999-4001]
Liu XL,Liu J,Guo AG,et al(2007).Influencing factors on wheat immature inflorescences culture.J Northwest A&F Univ,35(2):79-82(in Chinese with English abstract)[刘香利,刘缙,郭蔼光等(2007).小麦幼穗的离体培养及其影响因素研究.西北农林科技大学学报(自然科学版),35(2):79-82]
Ma L,Li Y,Chen Y,et al(2016).Improved drought and salt tolerance of Arabidopsis thaliana by ectopic expression of a cotton(Gossypium hirsutum)CBF gene.Plant Cell Tiss Org Cult,124(3):583-598
Meng L,Mao PC,Zhang XY,et al(2013).Somatic cell karyotypes of three Elytrigia elongate accessions.Acta Agrect Sin,21(4):821-827(in Chinese with English abstract)[孟林,毛培春,张晓燕等(2013).3份长穗偃麦草种质的体细胞核型分析.草地学报,21(4):821-827]
Meng L,Shang CY,Mao PC,et al(2009).A comprehensive evaluation of salt tolerance for germplasm and materials of Elytrigia at the seedling stage.Acta Pratacult Sin,18(4):67-74(in Chinese with English abstract)[孟林,尚春艳,毛培春等(2009).偃麦草属植物种质材料苗期耐盐性综合评价.草业学报,18(4):67-74]
Meng L,Yang HX,Mao PC,et al(2011).Assessment of interspecies drought resistance of Elytrigiaat the seedling stage.Acta Pratacult Sin,20(5):67-74(in Chinese with English abstract)[孟林,杨宏新,毛培春等(2011).偃麦草属植物种间苗期抗旱性评价.草业学报,20(5):67-74]
Meng L,Zhang L,Guo Q,et al(2016).Cloning and transformation of EeHKT1;4 gene from Elytrigia elongata.Protein Peptide Lett,23(5):488-494
Mujib A,Tonk D,Ali M(2014).Plant regeneration from protoplasts in Indian local Coriandrum sativum L.:scanning electron microscopy and histological evidences for somatic embryogenesis.Plant Cell Tiss Org Cult,117(3):323-334
Qiong YJ,Gui R,Lang ML(2011).Cloning and sequence analysis of DREB-like gene EeA P2.2 in Thinopyrum ponticum.J Plant Genet Resour,12(5):764-769(in Chinese with English abstract)[默韶京,桂茹,郎明林(2011).长穗偃麦草DREB类基因EeAP2.2的克隆与序列分析.植物遗传资源学报,12(5):764-769]
Qu RD,Chen Y(1983).A preliminary research on the function of enhancement of callus induction frequency by cold pretreatment in rice anther culture.Plant Physiol J,9(4):375-381(in Chinese with English abstract)[渠荣达,陈英(1983).低温预处理提高水稻花粉愈伤组织诱导频率的作用.植物生理学报,9(4):375-381]
Sahrawat A K,Chand S(2004).High frequency plant regeneration from coleoptile tissue of barley(Hordeum vulgare L.).Plant Sci,167(1):27-34
Wang JR,Yan ZH,Wei YM,et al(2004).Identification and molecular cloning of y-type high-molecular-weight glutenin subunit genes from Elytrigia elongate(Host)Nevski.Chin J Agric Biotechol,12(2):143-146(in Chinese with English abstract)[王际睿,颜泽洪,魏育明等(2004).长穗偃麦草Y型高分子量谷蛋白基因的鉴定与分子克隆.农业生物技术学报,12(2):143-146]
Wu GS,Hu YL,Song FP,et al(2006).Study on callus induction and plantlet regeneration of creeping bentgrass(Agrostis palustris Huds.).Biotechnolo Bull,4(22):16-21(in Chinese with English abstract)[吴桂胜,胡鸢雷,宋福平等(2006).剪股颖愈伤组织诱导与植株再生.生物技术通报,4(22):16-21]
Zhang LL,Chen SQ,Li HF,et al(2016).Development of wheat-Thinopyrum elongatum translocation lines resistant to Fusariumhead blight.Sci Agric Sin,49(18):3477-3488(in Chinese with English abstract)[张璐璐,陈士强,李海凤等(2016).小麦-长穗偃麦草7E抗赤霉病易位系培育.中国农业科学,49(18):3477-3488]
Zhang WJ,Yang CH,Liu F,et al(2010).The effects of different phytohormone combinations on callus induction of Poapratensis L.J Sichuan Agr Univ,28(2):187-190(in Chinese with English abstract)[张文君,杨春华,刘帆等(2010).不同植物生长调节物质配比对草地早熟禾愈伤组织形成的影响.四川农业大学学报,28(2):187-190]
Chen LP,Xue MF,Chen LZ,et al(2016).Preliminary study on regeneration plantlet from matured embryo of barnyardgrass by callus tissue culture.Plant Physiol J,52(12):1915-1920(in Chinese with English abstract)[陈丽萍,徐明飞,陈列忠等(2016).稗草成熟胚诱导愈伤组织再生植株的初步研究.植物生理学报,52(12):1915-1920]
Fan SH,Guo AG(2000).A study on the origin of HMW-GS 14 and 15 in Xiaoyan 6.J Northwest Agric Univ,28(6):1-5(in Chinese with English abstract)[范三红,郭蔼光(2000).小偃麦6号高分子质量麦谷蛋白14和15亚基来源分析.西北农业大学学报,28(6):1-5]
Gao SQ,Wang YB,Tang YM,et al(2011).Preliminary function research of EeNAC9 gene from Elytrigia elongata.Biotechnol Bull,(6):47-52(in Chinese with English abstract)[高世庆,王永波,唐益苗等(2011).长穗偃麦草EeNAC9基因功能初步研究.生物技术通报,(6):47-52]
Guo Q,Meng L,Mao PC,et al(2015).Salt tolerance in two tall wheatgrass species is associated with selective capacity for K+over Na+.Acta Physiol Plant,17081(37):1708-1760
Guo XY,Li HS,Peng KQ,et al(2011).Tissue culture and rapid propagation technology of Triathena lutarioriparia L.Liu sp.nov.Plant Physiol J,47(10):987-990(in Chinese with English abstract)[郭夏宇,李合松,彭克勤等(2011).南荻的组织培养与快速繁殖技术.植物生理学报,47(10):987-990]
He Y,Tian ZH,Zheng YL(2005).High frequency plant regeneration from mature embryos in vitro culture of tall fescue.Pratacult Sci,22(6):28-34(in Chinese with English abstract)[何勇,田志宏,郑用琏(2005).高羊茅成熟胚离体培养及高频植株再生.草业科学,22(6):28-34]
Huang K,Tang J,Liu ZZ,et al(2015).Influences of auxin and cytokine on propagation in vitro of Mulberry.J Southwest Univ,37(3):28-34(in Chinese with English abstract)[黄科,唐婧,刘自震等(2015).生长素和细胞分裂素对桑树离体繁育的影响.西南大学学报(自然科学版),37(3):28-34]
Huang W,Fu L,Li C,et al(2017).Quercetin,hyperin,and chlorogenicacid improve endothelial function by antioxidant,anti in flammatory,and ACE inhibitory effects.JFood Sci,82(5):1239-1246
Huo XW,Wei JH,Zhang H,et al(2004).Study of plant regeneration in wheatgrass(Agropyron Gaertn).Acta Agric Boreali Sin,19(1):17-20(in Chinese with English abstract)[霍秀文,魏建华,张辉等(2004).冰草属植物组织培养再生体系的建立.华北农学报,19(1):17-20]
Hussein Z,Dryanova A,Maret D,et al(2014).Gene expression analysis in the roots of salt-stressed wheat and the cytogenetic derivatives of wheat combined with the salt-tolerant wheatgrass,Lophopyrum elongatum.Plant Cell Rep,33(1):189-201
Jiang BN,Wang C,Pan JW(2014).Molecular regulatory mechanisms of hypocotyl elongation and phototropism in Arabidopsis.Plant Physiol J,50(10):1435-1444(in Chinese with English abstract)[姜楠,王超,潘建伟(1993).拟南芥下胚轴伸长与向光性的分子调控机理.植物生理学报,50(10):1435-1444]
Li WJ,Li XQ,Jia MM,et al(2012).Effects of 6-BA and 2,4-D on callus induction,growth and plantlet regeneration of shepherd’s purse[Capsella bursa-pastoris(L.)Medic].Plant Physiol J,48(2):141-146(in Chinese with English abstract)[李文静,李学强,贾毛毛等(2012).6-BA、NAA和2,4-D不同配比对荠菜愈伤组织诱导、生长及植株再生的影响.植物生理学报,48(2):141-146]
Li YJ,Liu JZ,Li B,et al(2010).Chromosomal location of the genes conferring the tolerance to phosphorus deficiency stress in Lophopyrum elongatum genome.J Genet Genomics,26(6):703-710(in Chinese with English abstract)[李玉京,刘建中,李滨等(2010).长穗偃麦草基因组中与耐低磷营养胁迫有关的基因的染色体定位.遗传学报,26(6):703-710]
Liu SY,Wang PW,Guan SY,et al(2013).Prelininary study on effects of 6-BA on callus induction of soybean.Hubei Agr Sci,52(16):3999-4001(in Chinese with English abstract)[刘思言,王丕武,关淑艳等(2013).6-BA对大豆愈伤组织诱导影响的初步研究.湖北农业科学,52(16):3999-4001]
Liu XL,Liu J,Guo AG,et al(2007).Influencing factors on wheat immature inflorescences culture.J Northwest A&F Univ,35(2):79-82(in Chinese with English abstract)[刘香利,刘缙,郭蔼光等(2007).小麦幼穗的离体培养及其影响因素研究.西北农林科技大学学报(自然科学版),35(2):79-82]
Ma L,Li Y,Chen Y,et al(2016).Improved drought and salt tolerance of Arabidopsis thaliana by ectopic expression of a cotton(Gossypium hirsutum)CBF gene.Plant Cell Tiss Org Cult,124(3):583-598
Meng L,Mao PC,Zhang XY,et al(2013).Somatic cell karyotypes of three Elytrigia elongate accessions.Acta Agrect Sin,21(4):821-827(in Chinese with English abstract)[孟林,毛培春,张晓燕等(2013).3份长穗偃麦草种质的体细胞核型分析.草地学报,21(4):821-827]
Meng L,Shang CY,Mao PC,et al(2009).A comprehensive evaluation of salt tolerance for germplasm and materials of Elytrigia at the seedling stage.Acta Pratacult Sin,18(4):67-74(in Chinese with English abstract)[孟林,尚春艳,毛培春等(2009).偃麦草属植物种质材料苗期耐盐性综合评价.草业学报,18(4):67-74]
Meng L,Yang HX,Mao PC,et al(2011).Assessment of interspecies drought resistance of Elytrigiaat the seedling stage.Acta Pratacult Sin,20(5):67-74(in Chinese with English abstract)[孟林,杨宏新,毛培春等(2011).偃麦草属植物种间苗期抗旱性评价.草业学报,20(5):67-74]
Meng L,Zhang L,Guo Q,et al(2016).Cloning and transformation of EeHKT1;4 gene from Elytrigia elongata.Protein Peptide Lett,23(5):488-494
Mujib A,Tonk D,Ali M(2014).Plant regeneration from protoplasts in Indian local Coriandrum sativum L.:scanning electron microscopy and histological evidences for somatic embryogenesis.Plant Cell Tiss Org Cult,117(3):323-334
Qiong YJ,Gui R,Lang ML(2011).Cloning and sequence analysis of DREB-like gene EeA P2.2 in Thinopyrum ponticum.J Plant Genet Resour,12(5):764-769(in Chinese with English abstract)[默韶京,桂茹,郎明林(2011).长穗偃麦草DREB类基因EeAP2.2的克隆与序列分析.植物遗传资源学报,12(5):764-769]
Qu RD,Chen Y(1983).A preliminary research on the function of enhancement of callus induction frequency by cold pretreatment in rice anther culture.Plant Physiol J,9(4):375-381(in Chinese with English abstract)[渠荣达,陈英(1983).低温预处理提高水稻花粉愈伤组织诱导频率的作用.植物生理学报,9(4):375-381]
Sahrawat A K,Chand S(2004).High frequency plant regeneration from coleoptile tissue of barley(Hordeum vulgare L.).Plant Sci,167(1):27-34
Wang JR,Yan ZH,Wei YM,et al(2004).Identification and molecular cloning of y-type high-molecular-weight glutenin subunit genes from Elytrigia elongate(Host)Nevski.Chin J Agric Biotechol,12(2):143-146(in Chinese with English abstract)[王际睿,颜泽洪,魏育明等(2004).长穗偃麦草Y型高分子量谷蛋白基因的鉴定与分子克隆.农业生物技术学报,12(2):143-146]
Wu GS,Hu YL,Song FP,et al(2006).Study on callus induction and plantlet regeneration of creeping bentgrass(Agrostis palustris Huds.).Biotechnolo Bull,4(22):16-21(in Chinese with English abstract)[吴桂胜,胡鸢雷,宋福平等(2006).剪股颖愈伤组织诱导与植株再生.生物技术通报,4(22):16-21]
Zhang LL,Chen SQ,Li HF,et al(2016).Development of wheat-Thinopyrum elongatum translocation lines resistant to Fusariumhead blight.Sci Agric Sin,49(18):3477-3488(in Chinese with English abstract)[张璐璐,陈士强,李海凤等(2016).小麦-长穗偃麦草7E抗赤霉病易位系培育.中国农业科学,49(18):3477-3488]
Zhang WJ,Yang CH,Liu F,et al(2010).The effects of different phytohormone combinations on callus induction of Poapratensis L.J Sichuan Agr Univ,28(2):187-190(in Chinese with English abstract)[张文君,杨春华,刘帆等(2010).不同植物生长调节物质配比对草地早熟禾愈伤组织形成的影响.四川农业大学学报,28(2):187-190]