黑果枸杞离体快繁技术研究
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摘要
【目的】探讨黑果枸杞的组织培养技术,为实现规模化高效离体快繁提供技术依据。【方法】以黑果枸杞当年生嫩茎为外植体,研究激素组合对黑果枸杞组培苗初代、增殖以及生根培养的影响。【结果】适合黑果枸杞初代培养的培养基为MS+6-BA1. 0 mg/L+NAA0. 2 mg/L,腋芽生长良好,萌芽率达88. 61%,且玻璃化情况较少;适合微枝增殖培养的培养基为MS+6-BA0. 1mg/L+NAA0. 2mg/L,微枝生长速度快,增殖倍数可达4. 8倍,少见玻璃化现象,且苗生长健壮,叶片嫩绿;适合微枝生根的培养基为1/2MS+IBA0. 2mg/·L,生根率达98%。适宜的移栽基质为营养土∶河沙∶蛭石=3∶1∶1(体积比)。【结论】黑果枸杞嫩茎离体培养和茎芽增殖可以获得再生植株实现离体快繁,本研究结果为黑果枸杞优良种质规模化繁殖提供了技术支持。
[Objective] To develop tissue culture technique for the Lycium ruthenicum,and provide technical basis for large scale and efficient in vitro rapid propagation. [Method]In this study,using tender stem of L. ruthenicum as explant,the effects of different factors on primary culture,subculture and rooting culture were studied. [Results]The results showed that: the suitable medium for axillary bud sprouting was MS + 6-BA1. 0 mg·L~(-1)+ NAA0. 2 mg·L~(-1),in which the axillary bud germination rate reached 88. 61%and the vitrification were rarely. The suitable medium for proliferation was MS + 6-BA 0. 1 mg·L~(-1)+ NAA0. 2 mg·L~(-1),in which the plantlets grew quickly. The proliferation rate was 4. 8 times,and there was no vitrification with robust seedling. The suitable medium for plantlet rooting was MS + IBA0. 2 mg·L~(-1),in which the rooting rate reached 98%. Suitable transplanting substrates were nutritional soil : river sand : vermiculite = 2: 1: 1. [Conclusion]The in vitro culture and stem bud subculture of Lycium ruthenicum can be used to obtain the in vitro rapid propagation of the regenerated plants. The results provide technical support for the large-scale propagation of the fine germplasm of Lycium ruthenicum.
[Objective] To develop tissue culture technique for the Lycium ruthenicum,and provide technical basis for large scale and efficient in vitro rapid propagation. [Method]In this study,using tender stem of L. ruthenicum as explant,the effects of different factors on primary culture,subculture and rooting culture were studied. [Results]The results showed that: the suitable medium for axillary bud sprouting was MS + 6-BA1. 0 mg·L~(-1)+ NAA0. 2 mg·L~(-1),in which the axillary bud germination rate reached 88. 61%and the vitrification were rarely. The suitable medium for proliferation was MS + 6-BA 0. 1 mg·L~(-1)+ NAA0. 2 mg·L~(-1),in which the plantlets grew quickly. The proliferation rate was 4. 8 times,and there was no vitrification with robust seedling. The suitable medium for plantlet rooting was MS + IBA0. 2 mg·L~(-1),in which the rooting rate reached 98%. Suitable transplanting substrates were nutritional soil : river sand : vermiculite = 2: 1: 1. [Conclusion]The in vitro culture and stem bud subculture of Lycium ruthenicum can be used to obtain the in vitro rapid propagation of the regenerated plants. The results provide technical support for the large-scale propagation of the fine germplasm of Lycium ruthenicum.
引文
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[5]章英才,张晋宁.两种盐浓度环境中的黑果枸杞叶的形态结构特征研究[J].宁夏大学学报(自然科学版),2004(4):365-367.ZHANG Y C,ZHANG J N. Study on the morphological structure ofleaves of Lycium ruthenicum[J]. Journal of Ningxia University(Natural Science),2004(4):365-367.
[6]鲁海,薛红霞,安君.枸杞组织培养及试管快繁试验初报[J].内蒙古林业,2002(7):32.LU H,XUE H X,AN J. Tissue culture and rapid growth test of Ly-cium barbarum[J]. Inner Mongolia Forestry,2002(7):32.
[7]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(6):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.
[8]田惠桥.枸杞茎尖培养[J].植物生理学通讯,1983(6):39.TIAN H Q. Culture of stem tip of Lycium barbarum[J]. PlantPhysiology Communications,1983(6):39.
[9]沈慧,米永伟,王龙强.外源硅对盐胁迫下黑果枸杞幼苗生理特性的影响[J].草地学报,2012,20(3):553-558.SHEN H,MI Y W,WANG L Q. Effects of exogenous silicon onphysiological characteristics of Lycium barbarum seedling under saltstress[J]. Acta Agrestia Sinica,2012,20(3):553-558.
[10] LV X P,WANG C J,CHENG Y,et al. Isolation and structuralcharacterization of a polysaccharide LRP4-A from Lycium rutheni-cum Murr[J]. Carbohydrate Research,2012,365(2):20-25.
[11]阿力同·其米克,王青锋,等.新疆产药用植物黑果枸杞遗传多样性的ISSR分析[J].植物科学学报,2013,31(5):517-524.A L T,WANG Q F,et al. ISSA Analysis on genetic diversity ofmedically important Lycium ruthenicum Murr. in Xinjiang[J].Plant Science Journal,2013,31(5):517-524.
[12]刘克彪,李爱德,李发明.四种生长调节剂对黑果枸杞嫩枝扦插成苗的影响[J].经济林研究,2014,32(3):99-103.LIU K B,LI A D,LI F M. Effects of four kinds of growth regula-tors softwood on cuttings seedling in Lycium ruthenicium[J]. Non-wood Forest Research,2014,32(3):99-103.
[13]马金平,李建国,王孝,等.黑果枸杞苗木快速繁育及建园技术[J].北方园艺,2013(9):185-187.MA J P,LI J G,WANG X,et al. Rapid breeding of Lycium ru-thenicium seedling and its gardening technology[J]. The NorthGarden,2013(9):185-187.
[14]葛红娟,梁美霞,戴洪义.玻璃化与正常苹果试管苗的叶片和茎的显微结构比较[J].植物生理学通讯,2010,46(3):223-227.GA H J,LIANG M X,DAI H Y. Comparison of micro-struc-tures of leaves and stems between vitrified and normal apple(Ma-lus pumila Mill.)plantlets in vitro[J]. Plant Physiology Commu-nications,2010,46(3):223-227.
[15]韩晶.‘月光”等5个枣品种的花药离体培养[D].石家庄:河北农业大学,2014.HAN J. In vitro anther culture of five Chinese jujube cultivars in-cluding‘Yueguang’[D]. Shijiazhuang:Agricultural Universityof Hebei,2014.
[16]高弘扬,夏秀英,张静雪,等.石竹玻璃化苗的显微结构及气孔运动特性[J].植物生理学报,2015,51(5):686-694.GAO H Y,XIA X Y,ZHANG J X,et al. Microstructure and sto-mate movement characteristic of Hyperhydric Pink(Dianthus chinen-sis L.)[J]. Plant Physiology Journal,2015,51(5):686-694.
[17]张瑞熙,申敏,殷家明,等.农杆菌介导转化油菜中几种影响玻璃化频率的因素[J].西南大学学报(自然科学版),2012,34(4):9-13.ZHANG R X,SHEN M,YIN J M,et al. Some factors affectingvitrification in brassicanapus transformation mediated by a grobac-terium tumefaciens[J]. Journal of Southwest University(NaturalScience),2012,34(4):9-13.
[2]矫晓丽,迟晓峰,董琦,等.柴达木野生黑果枸杞营养成分分析[J].氨基酸和生物源,2011,33(3):60-62.JIAO X L,CHI X F,DONG Q,et al. Analysis of nutrient compo-sition of Lycium ruthenicum of qaidam[J]. Amin Acids&BioticResources,2011,33(3):60-62.
[3]马继雄.道地药材黑果枸杞的应用研究进展及青海的发展前景[J].青海师范大学学报(自然科学版),2012,28(3):53-56.MA J X. The fruit of Lycium ruthenicum the application of the re-search development and prospect of the development of QinghaiProvince[J]. Journal of Qinghai Normal University(Natural Sci-ence),2012,28(3):53-56.
[4]陈红军,侯旭杰,白红进,等.黑果枸杞中的几种营养成分的分析[J].中国野生植物资源,2002(2):55.CHEN H J,HOU X J,BAI H J,et al. Analysis of several nutrientsin Lycium ruthenicum[J]. China’s Wild Plant Resources,2002(2):55.
[5]章英才,张晋宁.两种盐浓度环境中的黑果枸杞叶的形态结构特征研究[J].宁夏大学学报(自然科学版),2004(4):365-367.ZHANG Y C,ZHANG J N. Study on the morphological structure ofleaves of Lycium ruthenicum[J]. Journal of Ningxia University(Natural Science),2004(4):365-367.
[6]鲁海,薛红霞,安君.枸杞组织培养及试管快繁试验初报[J].内蒙古林业,2002(7):32.LU H,XUE H X,AN J. Tissue culture and rapid growth test of Ly-cium barbarum[J]. Inner Mongolia Forestry,2002(7):32.
[7]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(6):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.
[8]田惠桥.枸杞茎尖培养[J].植物生理学通讯,1983(6):39.TIAN H Q. Culture of stem tip of Lycium barbarum[J]. PlantPhysiology Communications,1983(6):39.
[9]沈慧,米永伟,王龙强.外源硅对盐胁迫下黑果枸杞幼苗生理特性的影响[J].草地学报,2012,20(3):553-558.SHEN H,MI Y W,WANG L Q. Effects of exogenous silicon onphysiological characteristics of Lycium barbarum seedling under saltstress[J]. Acta Agrestia Sinica,2012,20(3):553-558.
[10] LV X P,WANG C J,CHENG Y,et al. Isolation and structuralcharacterization of a polysaccharide LRP4-A from Lycium rutheni-cum Murr[J]. Carbohydrate Research,2012,365(2):20-25.
[11]阿力同·其米克,王青锋,等.新疆产药用植物黑果枸杞遗传多样性的ISSR分析[J].植物科学学报,2013,31(5):517-524.A L T,WANG Q F,et al. ISSA Analysis on genetic diversity ofmedically important Lycium ruthenicum Murr. in Xinjiang[J].Plant Science Journal,2013,31(5):517-524.
[12]刘克彪,李爱德,李发明.四种生长调节剂对黑果枸杞嫩枝扦插成苗的影响[J].经济林研究,2014,32(3):99-103.LIU K B,LI A D,LI F M. Effects of four kinds of growth regula-tors softwood on cuttings seedling in Lycium ruthenicium[J]. Non-wood Forest Research,2014,32(3):99-103.
[13]马金平,李建国,王孝,等.黑果枸杞苗木快速繁育及建园技术[J].北方园艺,2013(9):185-187.MA J P,LI J G,WANG X,et al. Rapid breeding of Lycium ru-thenicium seedling and its gardening technology[J]. The NorthGarden,2013(9):185-187.
[14]葛红娟,梁美霞,戴洪义.玻璃化与正常苹果试管苗的叶片和茎的显微结构比较[J].植物生理学通讯,2010,46(3):223-227.GA H J,LIANG M X,DAI H Y. Comparison of micro-struc-tures of leaves and stems between vitrified and normal apple(Ma-lus pumila Mill.)plantlets in vitro[J]. Plant Physiology Commu-nications,2010,46(3):223-227.
[15]韩晶.‘月光”等5个枣品种的花药离体培养[D].石家庄:河北农业大学,2014.HAN J. In vitro anther culture of five Chinese jujube cultivars in-cluding‘Yueguang’[D]. Shijiazhuang:Agricultural Universityof Hebei,2014.
[16]高弘扬,夏秀英,张静雪,等.石竹玻璃化苗的显微结构及气孔运动特性[J].植物生理学报,2015,51(5):686-694.GAO H Y,XIA X Y,ZHANG J X,et al. Microstructure and sto-mate movement characteristic of Hyperhydric Pink(Dianthus chinen-sis L.)[J]. Plant Physiology Journal,2015,51(5):686-694.
[17]张瑞熙,申敏,殷家明,等.农杆菌介导转化油菜中几种影响玻璃化频率的因素[J].西南大学学报(自然科学版),2012,34(4):9-13.ZHANG R X,SHEN M,YIN J M,et al. Some factors affectingvitrification in brassicanapus transformation mediated by a grobac-terium tumefaciens[J]. Journal of Southwest University(NaturalScience),2012,34(4):9-13.