降低黄芩再生苗玻璃化率及其生根的研究

doi:10.7525/j.issn.1673-5102.2016.01.013

摘要/Abstract

摘要： 为了降低黄芩组培苗的玻璃化率并提高其生根率，本研究以黄芩无菌苗茎段诱导的不定芽为实验材料，分别研究了6-苄基嘌呤（6-BA）、蔗糖、琼脂及多效唑（PP333）等组培条件对黄芩不定芽玻璃化的影响以及吲哚丁酸（IBA）对再生苗生根的影响。研究结果表明：低浓度的6-BA有利于降低不定芽的玻璃化率，当培养基中添加0.2 mg·L^-1 6-BA时，不定芽的玻璃化率最低且增殖系数比对照增加1倍。随着培养基中蔗糖浓度的升高，不定芽的玻璃化率显著降低，其增殖系数也有所降低。培养基中蔗糖浓度为25 g·L^-1时，黄芩不定芽长势最好且玻璃化率为零，增殖系数也最高。培养基中添加7.5 g·L^-1的琼脂，不定芽的玻璃化率较低，增殖系数也较高。培养基中添加多效唑（PP333）有利于缓解黄芩再生不定芽玻璃化状态，随着PP333的浓度增加，玻璃化率也逐渐降低。0.2 mg·L^-1 PP333对黄芩不定芽的壮苗起到了很好的作用，再生苗明显变得粗壮。0.1 mg·L^-1的IBA最有利于黄芩再生苗的生根，生根率为100%，平均生根数为7，移栽成活率达95%以上。

关键词: 黄芩, 不定芽, 玻璃化, 生根

Abstract: For reducing hyperhydric rate and improving rooting percentage of in vitro regenerated shoots from Scutellariae baicalensis Georgi, with the regenerated shoots induced from aseptic seedling stems, we studied the effect of N⁶-Benzyladenine(6-BA), sucrose, agar and paclobotrazol(PP333) on hyperhydricity of regenerated shoots in S.baicalensis and the effects of IBA on rooting of regenerated shoots. The hyperhydric rate significantly decreased with lower 6-BA concentration on the medium. The lowest hyperhydric rate and the highest multiplication rate were on the medium containing 0.2 mg·L^-1 6-BA. With the sucrose concentration increasing, the hyperhydric rate of S.baicalensis adventitious shoots was reduced, while their multiplication coefficient was also reduced. The best multiplication coefficient in vitro shoots without hyperhydricity were found on the medium containing 25 g·L^-1 sucrose. On the medium containing 7.5 g·L^-1 agar, the hyperhydric rate of S.baicalensis adventitious shoots was the lowest with higher proliferation coefficient. The medium containing PP333 positively affected the hyperhydric occurrence of regenerated shoots. With the increase of PP333 concentration on the medium, the hyperhydric rate was decreased gradually and in vitro shoots were stronger and healthier. The optimal rooting was got from the medium containing 0.1 mg·L^-1 IBA, with the rooting rate of 100% and seven roots per shoot. The survival rate of rooting plantlets was over 95% after transferred to soil.

Key words: Scutellariae baicalensis Georgi, adventitious shoots, hyperhydricity, rooting

韩淑兰；王慧梅*；张丹. 降低黄芩再生苗玻璃化率及其生根的研究[J]. 植物研究, 2016, 36(1): 90-96.

HAN Shu-Lan；WANG Hui-Mei*；ZHANG Dan. Decreasing the Hyperhydricity and Rooting of Scutellariae baicalensis Georgi[J]. Bulletin of Botanical Research, 2016, 36(1): 90-96.

参考文献

1.罗毓健,袁媛,黄璐琦.两来源黄芩再生体系培养条件的研究［J］.中国中医科学院中药研究所,2008,07(1):607-611.

2.Cao H B,Jiang Y J,Chen J J,et al.Arsenic accumulation in Scutellaria baicalensis Georgi and its effects on plant growth and pharmaceutical components［J］.Journal of Hazardous Materials,2009,171(06):508-513.

3.Mahmood N,Pizza C,Aquino R,et al.Inhibition of HIV infection by flavanoids［J］.Antiviral Res,1993,22:189-99.

4.张延红,高素芳,陈红刚,等.黄芩变异种质的生根壮苗研究［J］.中药材,2013,36(4):511-514.

5.Brearley T A,Vaidya B N,Joshee N.Cytokinin,Carbon Source,and Acclimatization Requirements for in Vitro Propagation of Scutellaria barbata D.Don and Scutellaria racemosa Pers［J］.American Journal of Plant Sciences,2014,5:3662-3672.

6.Ivanova M,Van Staden J.Influence of gelling agent and cytokinins on the control of hyperhydricity in Aloe polyphylla［J］.Plant Cell Tiss Organ Cult,2011,104:13-21.

7.Gaspar T,Kevers C,Bisbis B,et al.Loss of plant organogenic totipotency in the course of in vitro neoplastic progression［J］.In Vitro Cell Dev Biol Plant,2000,36(3):171-181.

8.Kataeva N V,Alexandrova I G,Raisa G,et al.Effect of applied and internal hormones on vitrification and apical necrosis of different plants cultured in vitro［J］.Plant Cell,Tissue and Culture,1991,27(2):149-154.

9.Perez-Tornero O,Egea J,Olmos E,et al.Control of hyperhydricity in micropropagated apricot cultivars［J］.In vitro cell Dev Biol.-plant,2001,37(2):250-254.

10.Ivanova M,van Staden J.Effect of ammanium ions and cytokinins on hyperhydricity and multiplication rate of in vitro regenerated shoots of Aloe polyphylla［J］.Plant Cell Tiss Organ Cult,2008,92:227-231.

11.Ziv M,Ariel T.Vitrification in relation to stomatal deformation and malfunction in carnation leaves in vitro［M］.//Debergh P C,Zimmerman R H.Micropropagation:technology and application.Kluwer Academic Publishers,Dordrecht,The Netherlands,1973:45-69.

12.Fujiwara K,Kozai T.Physical microenvironment and its effects［M］.//Aitken-Christie J,Kozai T,Smith M L,et al.Automation and Environmental Control in Plant Tissue Culture.Dordrecht:Kluwer Academic Publishers,1995:319-369.

13.吴晓玲,谢亚军,巫鹏举,等.植物生长调节剂对银柴胡细胞生长特性的影响［J］.植物研究,2005,25(2):173-176.

14.Smith M A,Spomer L A.Vessels,gels,liquid media,and support systems［M］.//AitkenChristie J,Kozai T,Smith M L,et al.Automation and Environmental Control in Plant Tissue Culture.Dordrecht:Kluwer Academic Publishers,1995:371-404.

15.王庭辉,马晖玲.和田苜蓿组织培养中玻璃化现象研究［J］.草原与草坪,2012,32(5):58-61.

16.王文静,李维强,王鹏.多效唑预处理对红金银花直接培养成壮苗的影响［J］.北方园艺,2012(09):184-185.

17.邢琳,李青.中国石竹试管苗玻璃化影响因素的研究［M］//.张启翔.中国观赏园艺研究进展.北京:中国林业出版社,2009:198-201.

18.Fracro F,Echeverrigaray S.Micropagation of Cunila galioides,a popular medicinal plant of South Brazil［J］.Plant Cell Tiss Org Cult,2001,64(1):1-4.

19.周书利,汤浩茹,代利娟,等.不同因素对草莓试管苗玻璃化的影响［J］.北方园艺,2013,(3):131-134.

20.Mayor M L,Nestares G,Zorzoli R,et al.Reduction of hyperhydricity in sunflower tissue culture［J］.Plant Cell,Tissue and Organ Culture,2003,72(1):99-103.

21.Debergh P,Aitken-Christie J,Cohen D,et al.Reconsideration of the term‘vitrification’as used in micropropagation［J］.Plant Cell Tissue Organ Cult,1992,30(2):135-140.

22.Brand M H.Agar and ammonium nitrate influence hyperhydricity,tissue nitrate and total nitrogen content of serviceberry(Amelanchier arborea) shoots in vitro［J］.Plant Cell,Tissue and Organ Culture,1993,35(3):203-209.

23.Casanova E,Moysset L,Trillas M I.Effects of agar concentration and vessel closure on the organogenesis and hyperhydricity of adventitious carnation shoots［J］.Biologia Plantarum,2008,52(1):1-8.

24.Wang Y L,Wang X D,Zhao B,et al.Reduction of hyperhydricity in the culture of Lepidium meyenii shoots by addition of rare earth elements［J］.Plant Growth Regul,2007,52(2):151-159.

25.吕复兵,朱根发.芦荟的组织培养与快繁技术［J］.北方园艺,2000(4):32-33.

26.王爱勤,何龙飞,裴润梅,等.组培条件对不同品种芦荟试管苗玻璃化的影响［J］.中国农学通报,2002,18(5):46-52.

27.Olmos E,Piqueras A,Martõ Ânez-Solano J R,et al.The subcellular localization of peroxidase and the implication of oxidative stress in hyperhydrated leaves of regenerated carnation plants［J］.Plant Sci,1997,130(1):97-105.

28.Krizan B,Ondrušiková E,Dradi G,et al.The effect of paclobutrazol on in vi tro rooting and growth of GF-677 hy brid peach rootstock［J］.Acta Physiologiae Plantarum,2006,28(1):21-26.

29.袁利利,张林,王厚新,等.华北五角枫‘京2’插穗生根过程中内源激素变化［J］.中国农学通报,2012,28(13):61-64.

30.Kesley D,Chaldecott M A.The role of endogenous auxin in root initiation.Ⅱ:Sensitivity,and evidence from studies on transgenic plant tissues［J］.Plant growth regulation,1993,13(1):77-84.

31.Van Der Krieken W M,Breteler H,Visser M,et a1.The role of the conversion of IBA into IAA on root regeneration in apple:Introduction of test system［J］.Plant Cell Rep,l993,12:203-206.

32.宋鹏飞,陈华江,姜燕琴,等.IBA对兔眼蓝浆果嫩枝扦插生根及内源激素变化的影响［J］.中国农学通报,2014,0(16):117-122.

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