日本野漆树的组织培养
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摘要
日本野漆树(Toxicodendron succedaneum)是一种独特的木本油料树种,以盛产漆蜡而闻名。本研究以带腋芽的茎段为外植体,通过筛选外植体消毒方法,控制褐变措施,以及腋芽启动、壮苗培养和生根诱导等适宜培养基,建立日本野漆树组织培养技术体系,从而为该树种良种的工厂化育苗提供技术支撑。结果表明:较适宜的外植体消毒方法为75%酒精浸泡10 s,2%洁尔灭浸泡1.5 min,0.1%升汞浸泡4 min。在MS培养基中添加0.8g·L~(-1) AC能有效地降低外植体的褐变。适宜的腋芽启动培养基为MS+2.0 mg·L~(-1) 6-BA+0.5 mg·L~(-1) NAA+0.8 g·L~(-1) AC。壮苗培养基为MS+1.0 mg·L~(-1) 6-BA+0.5 mg·L~(-1) ZT+0.1 mg·L~(-1) TDZ。培养基MS+0.1 mg·L~(-1) NAA+0.1mg·L~(-1) ZT+1.0 mg·L~(-1) 6-BA适用于愈伤组织诱导。培养基1/2MS+1.0 mg·L~(-1) GA_3+0.5 mg·L~(-1) NAA适用于生根诱导培养,生根率达到93.33%。
Toxicodendron succedaneum is a unique oil-producing woody species that is famous for producing lacquer wax. The stems with axillary buds from new sprout were collected as initial explants to screen the explants sterilization method, measure to prevent explants from browning, and suitable media components for axillary buds initiation, strong seedling culture, callus differentiation and root induction. The aim was to establish the technical system of tissue culture of T. succedaneum, and provide technical support for the industrialized seedling raising of its elite variety. The results showed that the suitable sterilization condition was to be successively soak in 75% ethanol for 10 s, 2% benzalkonium chloride for 1.5 min, and 0.1% mercuric chloride for 4 min. The addition of 0.8 g·L~(-1) activated carbon(AC) in the medium could effectively reduce the browning of explants. MS+2.0 mg·L~(-1) 6-BA+0.5 mg·L~(-1) NAA+0.8 g·L~(-1) AC was suitable to axillary bud initiation. MS+1.0 mg·L~(-1) 6-BA+0.5 mg·L~(-1) ZT+0.1 mg·L~(-1) TDZ was suitable to strong seedling culture. MS+0.1 mg·L~(-1) NAA+0.1 mg·L~(-1) ZT+1.0 mg·L~(-1) 6-BA and 1/2 MS+1.0 mg·L~(-1) GA3+0.5 mg·L~(-1) NAA was good at callus differentiation induction and root induction, respectively. The rooting rate reached 93.33%.
Toxicodendron succedaneum is a unique oil-producing woody species that is famous for producing lacquer wax. The stems with axillary buds from new sprout were collected as initial explants to screen the explants sterilization method, measure to prevent explants from browning, and suitable media components for axillary buds initiation, strong seedling culture, callus differentiation and root induction. The aim was to establish the technical system of tissue culture of T. succedaneum, and provide technical support for the industrialized seedling raising of its elite variety. The results showed that the suitable sterilization condition was to be successively soak in 75% ethanol for 10 s, 2% benzalkonium chloride for 1.5 min, and 0.1% mercuric chloride for 4 min. The addition of 0.8 g·L~(-1) activated carbon(AC) in the medium could effectively reduce the browning of explants. MS+2.0 mg·L~(-1) 6-BA+0.5 mg·L~(-1) NAA+0.8 g·L~(-1) AC was suitable to axillary bud initiation. MS+1.0 mg·L~(-1) 6-BA+0.5 mg·L~(-1) ZT+0.1 mg·L~(-1) TDZ was suitable to strong seedling culture. MS+0.1 mg·L~(-1) NAA+0.1 mg·L~(-1) ZT+1.0 mg·L~(-1) 6-BA and 1/2 MS+1.0 mg·L~(-1) GA3+0.5 mg·L~(-1) NAA was good at callus differentiation induction and root induction, respectively. The rooting rate reached 93.33%.
引文
Deng XA(1992).Cultivation of in vitro plantlets of lacquer tree in tissue culture.Guangxi Forest Sci,21:187-189(in Chinese without English abstract)[邓晓安(1992).漆树萌芽组织培养试管苗的培育.广西林业科技,21:187-189]
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Huang XZ,Jiang CF,Liao LL,et al(2006).Progress on molecular foundation of GA biosynthesis pathway and signaling.Chinese Bull Bot,23:499-510(in Chinese with English abstract)[黄先忠,蒋才富,廖立力等(2006).赤霉素作用机理的分子基础与调控模式研究进展.植物学通报,23:499-510]
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Luo YR,Han GW,Zhang X,et al(2017).Tissue culture of Toxicodendron vernicifluum.J Southwest For Univ,37:32-39(in Chinese with English abstract)[罗一然,韩国伟,张雪等(2017).小木漆组织培养技术研究.西南林业大学学报,37:32-39]
Thomas TD(2008).The role of activated charcoal in plant tissue culture.Biotechnol Adv,26:618-631
Wang DZ,Zhu ZR(1987).A preliminary report on tissue culture technology of lacquer tree.J Chin Lacquer,2:4-8(in Chinese with English abstract)[王道植,朱忠荣(1987).漆树组织培养技术研究初报.中国生漆,2:4-8]
Wang YR,Pan HY,Wu Y,et al(2014).Status and prospects of the utilization of lacquer tree resource.Furniture,35:27-31(in Chinese with English abstract)[王颖睿,潘海云,吴燕等(2014).漆树资源利用现状及发展前景.家具,35:27-31]
Yang SH(2011).Biological characteristics and prospect of development and utilization of Toxicodendron vernicifluum.Inner Mongolia Investiga Design,34:48-49(in Chinese without English abstract)[杨胜辉(2011).漆树的生物学特性及开发利用前景.内蒙古林业调查设计,34:48-49]
Zhang P,Liao SX,Cui K,et al(2013).Resources and species of Toxicodendron and its industry development prospects in China.World For Res,26:65-69(in Chinese with English abstract)[张鹏,廖声熙,崔凯等(2013).中国漆树资源与品种现状及产业发展前景.世界林业研究,26:65-69]
Zhang YL,Liu YJ,Huang JX,et al(2006).Tissue culture method of Anthurium andraeanum explants with entophytic microorganism.Southwest Hortic,34:9-11(in Chinese with English abstract)[张寅玲,刘艳军,黄俊轩等(2006).含内生菌外植体红掌组培方法的研究.西南园艺,34:9-11]
Zhao YM,Zhao YM,He CZ,et al(2015).Study on the prevention and cure of browning and contamination during tissue culture of Toxicodendron vernicifluum.Chin Agr Sci Bull,31:33-38(in Chinese with English abstract)[赵月明,赵雁鸣,何承忠等(2015).漆树组织培养中污染与褐变的防治.中国农学通报,31:33-38]
Zhao YY,Liu JY,Ma Z,et al(2017).Tissue culture and high-frequency regeneration of Lepidium sativum.Plant Physiol J,53:234-240(in Chinese with English abstract)[赵杨阳,刘佳钰,马征等(2017).家独行菜的组织培养与高频再生.植物生理学报,53:234-240]
Fu SY,Pan CM,Liu LT,et al(2015).Tissue culture and rapid propagation of medicinal plant Pithecellobium clypearia.Plant Physiol J,51:2195-2200(in Chinese with English abstract)[付姝颖,潘超美,刘良婷等(2015).药用植物猴耳环的组织培养与快速繁殖.植物生理学报,51:2195-2200]
Huang XZ,Jiang CF,Liao LL,et al(2006).Progress on molecular foundation of GA biosynthesis pathway and signaling.Chinese Bull Bot,23:499-510(in Chinese with English abstract)[黄先忠,蒋才富,廖立力等(2006).赤霉素作用机理的分子基础与调控模式研究进展.植物学通报,23:499-510]
Kim D H,Gopal J,Sivanesan I(2017).Nanomaterials in plant tissue culture:the disclosed and undisclosed.RSC Adv,7:36492-36505
Liu W,Xie BX,Yu JF,et al(2008).Extraction techniques of lacquer wax in Rhus succedanea.Nonwood Forest Res,26:58-61(in Chinese with English abstract)[刘伟,谢碧霞,余江帆等(2008).日本野漆树漆蜡萃取技术.经济林研究,26:58-61]
Luo YR,Han GW,Zhang X,et al(2017).Tissue culture of Toxicodendron vernicifluum.J Southwest For Univ,37:32-39(in Chinese with English abstract)[罗一然,韩国伟,张雪等(2017).小木漆组织培养技术研究.西南林业大学学报,37:32-39]
Thomas TD(2008).The role of activated charcoal in plant tissue culture.Biotechnol Adv,26:618-631
Wang DZ,Zhu ZR(1987).A preliminary report on tissue culture technology of lacquer tree.J Chin Lacquer,2:4-8(in Chinese with English abstract)[王道植,朱忠荣(1987).漆树组织培养技术研究初报.中国生漆,2:4-8]
Wang YR,Pan HY,Wu Y,et al(2014).Status and prospects of the utilization of lacquer tree resource.Furniture,35:27-31(in Chinese with English abstract)[王颖睿,潘海云,吴燕等(2014).漆树资源利用现状及发展前景.家具,35:27-31]
Yang SH(2011).Biological characteristics and prospect of development and utilization of Toxicodendron vernicifluum.Inner Mongolia Investiga Design,34:48-49(in Chinese without English abstract)[杨胜辉(2011).漆树的生物学特性及开发利用前景.内蒙古林业调查设计,34:48-49]
Zhang P,Liao SX,Cui K,et al(2013).Resources and species of Toxicodendron and its industry development prospects in China.World For Res,26:65-69(in Chinese with English abstract)[张鹏,廖声熙,崔凯等(2013).中国漆树资源与品种现状及产业发展前景.世界林业研究,26:65-69]
Zhang YL,Liu YJ,Huang JX,et al(2006).Tissue culture method of Anthurium andraeanum explants with entophytic microorganism.Southwest Hortic,34:9-11(in Chinese with English abstract)[张寅玲,刘艳军,黄俊轩等(2006).含内生菌外植体红掌组培方法的研究.西南园艺,34:9-11]
Zhao YM,Zhao YM,He CZ,et al(2015).Study on the prevention and cure of browning and contamination during tissue culture of Toxicodendron vernicifluum.Chin Agr Sci Bull,31:33-38(in Chinese with English abstract)[赵月明,赵雁鸣,何承忠等(2015).漆树组织培养中污染与褐变的防治.中国农学通报,31:33-38]
Zhao YY,Liu JY,Ma Z,et al(2017).Tissue culture and high-frequency regeneration of Lepidium sativum.Plant Physiol J,53:234-240(in Chinese with English abstract)[赵杨阳,刘佳钰,马征等(2017).家独行菜的组织培养与高频再生.植物生理学报,53:234-240]