兴安百里香茎芽增殖途径的植株再生
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摘要
为探索野生兴安百里香组织培养最佳培养条件获得完整再生植株,以五大连池野生兴安百里香为研究对象,研究外植体类型、消毒时间和培养基种类对兴安百里香茎芽增殖与微枝生根的影响。结果表明:带腋芽茎段为兴安百里香组培的最佳外植体,采用质量浓度0. 1%升汞浸泡4 min的方法消毒效果最佳;最适愈伤组织诱导培养基为MS+6-BA1. 0 mg/L+2,4-D0. 5 mg/L+蔗糖30 g/L+琼脂粉6 g/L;最适茎芽增殖培养基为MS+6-BA1. 0 mg/L+2,4-D0. 5 mg/L;最适微枝生根培养基为MS+6-BA 1. 0 mg/L+2,4-D 0. 8 mg/L。此研究结果为通过工厂化育苗技术培育野生兴安百里香优质观赏苗木奠定了基础。
In order to explore the optimal culture conditions for wild Thymus dahuricus tissue culture to obtain a complete regenerated plant,we studied the effects of explant type,disinfection time and medium variety on stem bud proliferation and microbranched rooting of Thymus dahuricus with Wudalianchi wild Thymus dahuricus. The results showed that: the best explants for tissue culture of Thymus dahuricus were axillary bud stem segment. The best disinfection effect was achieved by soaking for 4 min with a mass concentration of 0. 1% mercury. The best medium for callus induction was: MS + 6-BA1. 0 mg/L + 2,4-D0. 5 mg/L + saccharose 30 g/L + powdered agar 6 g/L. The most suitable medium for stem-bud multiplication was: MS + 6-BA1. 0 mg/L + 2,4-D0. 5 mg/L. The best medium for rooting of microtwigs was: MS + 6-BA 1. 0 mg/L + 2,4-D 0. 8 mg/L. The results laid a foundation for the cultivation of high quality ornamental seedlings of wild Thymus dahuricus by means of industrial seedling raising technology.
In order to explore the optimal culture conditions for wild Thymus dahuricus tissue culture to obtain a complete regenerated plant,we studied the effects of explant type,disinfection time and medium variety on stem bud proliferation and microbranched rooting of Thymus dahuricus with Wudalianchi wild Thymus dahuricus. The results showed that: the best explants for tissue culture of Thymus dahuricus were axillary bud stem segment. The best disinfection effect was achieved by soaking for 4 min with a mass concentration of 0. 1% mercury. The best medium for callus induction was: MS + 6-BA1. 0 mg/L + 2,4-D0. 5 mg/L + saccharose 30 g/L + powdered agar 6 g/L. The most suitable medium for stem-bud multiplication was: MS + 6-BA1. 0 mg/L + 2,4-D0. 5 mg/L. The best medium for rooting of microtwigs was: MS + 6-BA 1. 0 mg/L + 2,4-D 0. 8 mg/L. The results laid a foundation for the cultivation of high quality ornamental seedlings of wild Thymus dahuricus by means of industrial seedling raising technology.
引文
[1]赵波.辽宁省百里香属(Thymus L.)植物生物学特性及挥发油特性研究[D].沈阳:沈阳农业大学,2015.ZHAO B. Studies on biological and essential oil characteristics of Thymus L. in Liaoning Province[D]. Shenyang:Shenyang Agricultural University,2015.
[2]杨敏.百里香研究进展[J].园艺与种苗,2018,38(11):68-70.YANG M. Research advances on Thymus mongolicus[J]. Horticulture&Seed,2018,38(11):68-70.
[3]于二汝,王少铭,侯颖辉,等. 5种黔引百里香的形态特征和蒸馏法提取精油的化学型比较研究[J].热带作物学报,2018,39(1):84-92YU E R,WANG S M,HOU Y H,et al. Botanical characters and composition of essential oils from five varieties of Thyme grown in Guizhou area[J]. Chinese Journal of Tropical Crops,2018,39(1):84-92.
[4] SAIDI M,GHAFOURIAN S,ZARINABAADI M,et al. In vitro antimicrobial and antioxidant activity of black thyme(Thymbra spicata L.)essential oils[J]. Roum Archives of Microbiology and Immunology,2012,71(2):61-69.
[5] ZABOROWSKA Z A,PRZYGONSKI K,BILSKA A. Antioxidative effect of thyme(Thymus vulgaris)in sunflower oil[J]. Acta Sci.Pol.,Technol. Aliment,2012,11(3):283-291.
[6]吴爽,魏凤香,李红枝,等.柠檬百里香叶挥发油成分分析及对肝癌细胞毒性作用[J].中药材,2013,36(5):756-759.WU S,WEI F X,LI H Z,et al. Chemical composition of essential oil from Thymus citriodorus and its toxic effect on liver cancer cells[J]. Journal of Chinese Medicinal Materials,2013,36(5):756-759.
[7]刘光发,宋海燕,罗婉如,等.百里香-丁香罗勒精油抗菌纸对草莓的防腐保鲜效果[J],包装工程,2018,39(19):91-97.LIU G F,SONG H Y,LUO W R,et al. Effect of antimicrobial paper coated with Thymus vulgaris L. and Ocimum gratissimum L. essential oil on preservation of strawberry[J]. Packaging Engineering,2018,39(19):91-97.
[8]佟晶晶,王明阳,潘佳雯,等.植物精油抗菌和抗真菌活性的研究进展[J].中国饲料,2018,29(7):46-49.TONG J J,WANG M Y,PAN J W,et al. Research advances in antibacterial and antifungal activity of plant essential oils[J]. China Feed,2018,29(7):46-49.
[9]李晓东,王永飞,马三梅,等.东北百里香组培再生体系的建立普通百里香的组织培养和快速繁殖[J].湖北农业科学,2009,48(6):1294-1297.LI X D,WANG Y F,MA S M,et al. Tissue culture and rapid propagation of Thymus vulgaris[J]. Hubei Agricultural Sciences,2009,48(6):1294-1297.
[10]宋阳,王冲,魏岩.兴安百里香播种繁殖研究[J].种子,2014,33(1):116-118.SONG Y,WANG C,WEI Y. Study on characteristics of seeds and sowing of Thymus dahuricus Serg.[J]. Seed,2014,33(1):116-118.
[11]徐世千,李晓东,张建国.不同方法提取组培百里香精油质量及成分的比较分析[J].植物科学学报,2013,31(6):609-615.XU S Q,LI X D,ZHANG J G. Comparative analysis of essential oil quality and composition from tissue culture seedlings of Thymus vulgaris L using different extraction methods[J]. Plant Science Journal,2013,31(6):609-615.
[12]徐治朋.红花百里香组织培养体系优化及挥发油产量的调节研究[D].重庆:西南大学,2016.XU Z P. Study on optimization of tissue culture system and regulation of essential oil yield of Thymus serpyllum L.[D]. Chongqing:Southwest University,2016.
[13]王玲,杨丽鹏,张秀珍,等.东北百里香组培再生体系的建立[J].园艺学报,2011,38(6):1185-1190.WANG L,YANG L P,ZHANG X Z,et al. The establishment of the adventitious bud regeneration system of Thymus mandschuricus[J]. Acta Horticulturae Sinica,2011,38(6):1185-1190.
[14]赵庆臻,闫华超,程霜,等.地椒的组织培养及植株再生[J].植物生理学通讯,2002,38(4):357.ZHAO Q Z,YAN H C,CHENG S,et al. The tissue culture and plant regeneration of ground pepper[J]. Plant Physiology Communications,2002,38(4):357.
[15]李晓东,徐世千,鲁朝辉,等.苯丙氨酸和茉莉酸甲酯对百里香离体培养芽生长及其精油的影响[J].香料香精化妆品,2014,42(2):1-6.LI X D,XU S Q,LU Z H,et al. Study on the effects of phenylalanine and methyl jasmonate on profile of essential oil from tissue culture of Thymus vulgaris L[J]. Flavour Fragrance Cosmetics,2014,42(2):1-6.
[16]徐治朋,李晓东,鲁朝晖,等. Cu2+对离体培养的红花百里香不定芽生长及挥发油的影响[J].西南大学学报(自然科学版),2016,38(5):37-44.XU Z P,LI X D,LU Z H,et al. Effects of Cu(2+)on the growth of the adventitious buds of Thyme serpyllumin in vitro culture and their essential oil[J]. Journal of Southwest University(Natural Science Edition),2016,38(5):37-44.
[17]罗彩霞,田永强,潘彦彪.百里香研究进展[J].安徽农业科学,2014,42(33):11669-11672.LUO C X,TIAN Y Q,PAN Y B. Research progress of Thymus[J]. Journal of Anhui Agricultural Sciences,2014,42(33):11669-11672.
[18]穆丹,梁英辉.佳木斯地区百里香的引种栽培及园林应用研究[J].安徽农学通报,2013,19(16):31-32.MU D,LIANG Y H. Introduction and landscape application of Thyme in Jiamusi region[J]. Anhui Agricultural Science Bulletin,2013,19(16):31-32.
[19]李艳霞,刘忠玲,刘建明,等.树莓品种“波拉纳”组培繁殖体系建立[J].森林工程,2018,34(2):26-29.LI Y X,LIU Z L,LIU J M,et al. Establishment of tissue culture and propagation system for raspberry(Rubus idaeus)cultivar"Polana"[J]. Forest Engineering,2018,34(2):26-29.
[20]李琳,杜倩,梁素钰.黑果枸杞无菌苗愈伤组织诱导体系的建立[J].林业科技,2018,43(06):1-3.LI L,DU Q,LIANG S Y. Establishment of callus induction of non-bacterial seedling of lycium ruthenicum[J]. Forestry Science&Technology,2018,43(6):1-3.
[21]覃杰明,何含杰,张党权,等. 6-BA和GA3对盐胁迫下红杆铁皮石斛幼苗生理生化影响[J].亚热带植物科学,2016,45(1):27-31.QIN J M,HE H J,ZHANG D Q,et al. Physiological and biochemical influences of 6-BA and GA3 on red-stem dendrobium officinale under salt stress[J]. Subtropical Plant Science,2016,45(1):27-31.
[22]李学东,华帅,刘长安. NAA与6-BA对黑金丝柚木组织培养的影响[J].亚热带植物科学,2017,46(4):379-382.LI X D,HUA S,LIU C A,Effects of NAA and 6-BA on tissue culture of Tecton grandis[J]. Subtropical Plant Science,2017,46(4):379-382.
[23]张虎,巫建新,许建民,等. 6-BA与ZT对芫花茎段外植体组织培养的影响[J].江苏林业科技,2018,45(4):29-32.ZHANG H,WU J X,XU J M,et al. Effects of 6-BA and ZT on the tissue culture of Daphne genkwa segment explant[J]. Journal of Jiangsu Forestry Science&Technology,2018,45(4):29-32.
[2]杨敏.百里香研究进展[J].园艺与种苗,2018,38(11):68-70.YANG M. Research advances on Thymus mongolicus[J]. Horticulture&Seed,2018,38(11):68-70.
[3]于二汝,王少铭,侯颖辉,等. 5种黔引百里香的形态特征和蒸馏法提取精油的化学型比较研究[J].热带作物学报,2018,39(1):84-92YU E R,WANG S M,HOU Y H,et al. Botanical characters and composition of essential oils from five varieties of Thyme grown in Guizhou area[J]. Chinese Journal of Tropical Crops,2018,39(1):84-92.
[4] SAIDI M,GHAFOURIAN S,ZARINABAADI M,et al. In vitro antimicrobial and antioxidant activity of black thyme(Thymbra spicata L.)essential oils[J]. Roum Archives of Microbiology and Immunology,2012,71(2):61-69.
[5] ZABOROWSKA Z A,PRZYGONSKI K,BILSKA A. Antioxidative effect of thyme(Thymus vulgaris)in sunflower oil[J]. Acta Sci.Pol.,Technol. Aliment,2012,11(3):283-291.
[6]吴爽,魏凤香,李红枝,等.柠檬百里香叶挥发油成分分析及对肝癌细胞毒性作用[J].中药材,2013,36(5):756-759.WU S,WEI F X,LI H Z,et al. Chemical composition of essential oil from Thymus citriodorus and its toxic effect on liver cancer cells[J]. Journal of Chinese Medicinal Materials,2013,36(5):756-759.
[7]刘光发,宋海燕,罗婉如,等.百里香-丁香罗勒精油抗菌纸对草莓的防腐保鲜效果[J],包装工程,2018,39(19):91-97.LIU G F,SONG H Y,LUO W R,et al. Effect of antimicrobial paper coated with Thymus vulgaris L. and Ocimum gratissimum L. essential oil on preservation of strawberry[J]. Packaging Engineering,2018,39(19):91-97.
[8]佟晶晶,王明阳,潘佳雯,等.植物精油抗菌和抗真菌活性的研究进展[J].中国饲料,2018,29(7):46-49.TONG J J,WANG M Y,PAN J W,et al. Research advances in antibacterial and antifungal activity of plant essential oils[J]. China Feed,2018,29(7):46-49.
[9]李晓东,王永飞,马三梅,等.东北百里香组培再生体系的建立普通百里香的组织培养和快速繁殖[J].湖北农业科学,2009,48(6):1294-1297.LI X D,WANG Y F,MA S M,et al. Tissue culture and rapid propagation of Thymus vulgaris[J]. Hubei Agricultural Sciences,2009,48(6):1294-1297.
[10]宋阳,王冲,魏岩.兴安百里香播种繁殖研究[J].种子,2014,33(1):116-118.SONG Y,WANG C,WEI Y. Study on characteristics of seeds and sowing of Thymus dahuricus Serg.[J]. Seed,2014,33(1):116-118.
[11]徐世千,李晓东,张建国.不同方法提取组培百里香精油质量及成分的比较分析[J].植物科学学报,2013,31(6):609-615.XU S Q,LI X D,ZHANG J G. Comparative analysis of essential oil quality and composition from tissue culture seedlings of Thymus vulgaris L using different extraction methods[J]. Plant Science Journal,2013,31(6):609-615.
[12]徐治朋.红花百里香组织培养体系优化及挥发油产量的调节研究[D].重庆:西南大学,2016.XU Z P. Study on optimization of tissue culture system and regulation of essential oil yield of Thymus serpyllum L.[D]. Chongqing:Southwest University,2016.
[13]王玲,杨丽鹏,张秀珍,等.东北百里香组培再生体系的建立[J].园艺学报,2011,38(6):1185-1190.WANG L,YANG L P,ZHANG X Z,et al. The establishment of the adventitious bud regeneration system of Thymus mandschuricus[J]. Acta Horticulturae Sinica,2011,38(6):1185-1190.
[14]赵庆臻,闫华超,程霜,等.地椒的组织培养及植株再生[J].植物生理学通讯,2002,38(4):357.ZHAO Q Z,YAN H C,CHENG S,et al. The tissue culture and plant regeneration of ground pepper[J]. Plant Physiology Communications,2002,38(4):357.
[15]李晓东,徐世千,鲁朝辉,等.苯丙氨酸和茉莉酸甲酯对百里香离体培养芽生长及其精油的影响[J].香料香精化妆品,2014,42(2):1-6.LI X D,XU S Q,LU Z H,et al. Study on the effects of phenylalanine and methyl jasmonate on profile of essential oil from tissue culture of Thymus vulgaris L[J]. Flavour Fragrance Cosmetics,2014,42(2):1-6.
[16]徐治朋,李晓东,鲁朝晖,等. Cu2+对离体培养的红花百里香不定芽生长及挥发油的影响[J].西南大学学报(自然科学版),2016,38(5):37-44.XU Z P,LI X D,LU Z H,et al. Effects of Cu(2+)on the growth of the adventitious buds of Thyme serpyllumin in vitro culture and their essential oil[J]. Journal of Southwest University(Natural Science Edition),2016,38(5):37-44.
[17]罗彩霞,田永强,潘彦彪.百里香研究进展[J].安徽农业科学,2014,42(33):11669-11672.LUO C X,TIAN Y Q,PAN Y B. Research progress of Thymus[J]. Journal of Anhui Agricultural Sciences,2014,42(33):11669-11672.
[18]穆丹,梁英辉.佳木斯地区百里香的引种栽培及园林应用研究[J].安徽农学通报,2013,19(16):31-32.MU D,LIANG Y H. Introduction and landscape application of Thyme in Jiamusi region[J]. Anhui Agricultural Science Bulletin,2013,19(16):31-32.
[19]李艳霞,刘忠玲,刘建明,等.树莓品种“波拉纳”组培繁殖体系建立[J].森林工程,2018,34(2):26-29.LI Y X,LIU Z L,LIU J M,et al. Establishment of tissue culture and propagation system for raspberry(Rubus idaeus)cultivar"Polana"[J]. Forest Engineering,2018,34(2):26-29.
[20]李琳,杜倩,梁素钰.黑果枸杞无菌苗愈伤组织诱导体系的建立[J].林业科技,2018,43(06):1-3.LI L,DU Q,LIANG S Y. Establishment of callus induction of non-bacterial seedling of lycium ruthenicum[J]. Forestry Science&Technology,2018,43(6):1-3.
[21]覃杰明,何含杰,张党权,等. 6-BA和GA3对盐胁迫下红杆铁皮石斛幼苗生理生化影响[J].亚热带植物科学,2016,45(1):27-31.QIN J M,HE H J,ZHANG D Q,et al. Physiological and biochemical influences of 6-BA and GA3 on red-stem dendrobium officinale under salt stress[J]. Subtropical Plant Science,2016,45(1):27-31.
[22]李学东,华帅,刘长安. NAA与6-BA对黑金丝柚木组织培养的影响[J].亚热带植物科学,2017,46(4):379-382.LI X D,HUA S,LIU C A,Effects of NAA and 6-BA on tissue culture of Tecton grandis[J]. Subtropical Plant Science,2017,46(4):379-382.
[23]张虎,巫建新,许建民,等. 6-BA与ZT对芫花茎段外植体组织培养的影响[J].江苏林业科技,2018,45(4):29-32.ZHANG H,WU J X,XU J M,et al. Effects of 6-BA and ZT on the tissue culture of Daphne genkwa segment explant[J]. Journal of Jiangsu Forestry Science&Technology,2018,45(4):29-32.