Study on Rapid Seedling-raising Technology of Tree Peony Embryo Culture

doi:10.7525/j.issn.1673-5102.2017.05.008

Abstract

Abstract: This study used Paeonia ostii ‘Fengdan’ and Paeonia rockii as explants, established efficient method of Seedling-raising of tree peony embryo culture through the study of hormones, light conditions, medium ingredients and activated carbon(AC) on the in vitro embryo culture. The main result:In vitro embryos radicle germination in 10 d, grow true leaves in 20 d, root system developed in 25 d, transplanting in 40 d, after 3 months the seedlings grow well in the medium of improved MS(calcium doubled)+0.2 g·L^-1AC+0.5 mg·L^-1GA+0.5 mg·L^-1BA. Suitable medium for ‘Fengdan’:improved MS(calcium doubled,macroelement doubled)+0.6 g·L^-1AC+1.0 mg·L^-1GA; medium for P.rockii:improved MS(calcium doubled,macroelement doubled)+0.6 g·L^-1 AC+0.5 mg·L^-1GA; The seedling rate of ‘Fengdan’ was 63.88%, the survival rate was 66.34%, and the rate of seedling of P.rockii was 93.75%, and survival rates vary among breeds, with a maximum of 100%.

Key words: embryo culture, Paeonia ostii ‘Fengdan’, Paeonia rockii, activated carbon

CLC Number:

S567.1+5

XU Li, CHENG Fang-Yun, ZHONG Yuan. Study on Rapid Seedling-raising Technology of Tree Peony Embryo Culture[J]. Bulletin of Botanical Research, 2017, 37(5): 690-699.

References

1. Li S S,Yuan R Y,Chen L G,et al. Systematic qualitative and quantitative assessment of fatty acids in the seeds of 60 tree peony (Paeonia section Moutan DC.) cultivars by GC-MS[J]. Food Chemistry,2015,173:133-140.
2. 李嘉珏. 中国牡丹[M]. 北京:中国大百科全书出版社,2011. Li J J. Tree Peony of China[M]. Beijing:China Encyclopedia Publishing House,2011.
3. 成仿云,杜秀娟. 低温与赤霉素处理对‘凤丹’牡丹种子萌发和幼苗生长的影响[J]. 园艺学报,2008,35(4):553-558. Cheng F Y,Du X J. Effects of Chilling and Gibberellic Acid on the Seed Germination and Seed-ling Growth in Paeoniaostii ‘FengDan’[J]. Acta Horticulturae Sinica,2008,35(4):553-558.
4. 成仿云,李嘉珏,陈德忠,等. 中国紫斑牡丹[M]. 北京:中国林业出版社,2005:77-84. Cheng F Y,Li J J,Chen D Z,et al. Chinese Flare Mudan[M]. Beijing:China Forestry Publishing House,2005:77-84.
5. 潘晓,陈瑞丹. ‘淡丰后’梅胚培养影响因素的研究[J]. 北京林业大学学报,2010,2(32):88-92. Pan X,Chen R D. Factors affecting embryo culture of Prunusmume ‘Dan Fenghou’[J]. Journal of Beijing Forest University,2010,2(32):88-92.
6. Shen M M,Tang Z J,Da Silva Jat,et al. Induction and proliferation of axillary shoots from in vitro culture of Paeonia lactiflora Pall. mature zygotic embryos[J]. New Zealand Journal of Crop and Horticultural Science,2015,43(1):42-52.
7. 杜学梅,李登科,王永康,等. 枣胚培技术体系的建立[J]. 园艺学报,2005,32(3):496-499. Du X M,Li D K,Wang Y K,et al. Estabilishment of Embryo Culture Technical System of Chinese Jujube (Zizyphus jujube Mill.)[J]. ActaHorticulturae sinica,2005,32(3):496-499.
8. 王际轩,李淑珍,李博文,等. 山楂砧木苗的种胚培养[J]. 园艺学报,1982,9(4):7-10. Wang J X,Li S Z,Li B W,et al. embryo culture of Hawthorn (Crataegus pinnatifida) rootstocks[J]. Acta Horticulturae Sinica,1982,9(4):7-10.
9. 王元裕,周碧英,赵安祥,等. 柑桔胚培养技术的研究Ⅰ,-成熟种子胚的分离培养[J]. 园艺学报,1979,6(1):3-8,35. Wang Y Y,Zhou B Y,Zhao A X,et al. Studies on the technique of citrus embryo culture Ⅰ,The segregate culture of citrus embryos of mature seed[J]. ActaHorticulturae sinica,1979,6(1):3-8,35.
10. Zilis M R,Meyer M M. Rapid in vitro germination of immature,dormant embryos[J]. Comb ProcAnnu Meet Int Plant Propag Soc,1976.
11. Buchheim J A T,Burkhart L F,Meyer M M Jr. Effect of exogenous gibberellic acid,abscisic acid,and benzylaminopurine on epicotyl dormancy of cultured herbaceous peony embryos[J]. Plant cell,Tissue and Organ Culture,1994,36(1):35-43.
12. Wang H,Van Stadenj. Establishment of in vitro cultures of tree Peonies[J]. South African Journal of Botany,2001,67(2):358-361.
13. Brukhin V B,Batygina T B. Embryo culture and somatic embryogenesis in culture of Paeonia anomala[J]. Phytomorphology,1994,44(34):151-157.
14. 刘会超,贾文庆,王坤. 牡丹胚培养及丛生苗继代培养研究[J]. 北方园艺,2010,(6):172-174. Liu H C,Jia W Q,Wang K. The Culture of Peony Embryo and Subculture of Multiple Bud[J]. Northern Horticulture,2010,(6):172-174.
15. 王莹,何桂梅,韩丽晓. 紫斑牡丹胚培养及幼苗生长的研究[J]. 湖南农业科学,2012(9):103-106. Wang Y,He G M,Han L X. Study on Embryo-culture and Seedling Growth for Paeonia rockii[J]. Hunan Agricultural Sciences,2012(9):103-106.
16. Jana S,Sivanesan I,Lim M Y,et al. In vitro zygotic embryo germination and somatic embryogenesis through cotyledonary explants of Paeonia lactiflora Pall[J]. Flower Research Journal,2013,21(1):17-22.
17. James A C,Harris R A,Mantell S H. Paeonia Species (Tree Peonies)[M]. //Bajaj Y P S. Trees IV. Berlin Heidelberg:Springer,1996:244-268.
18. Fridborg G,Pedersen M,Landstrom L E,et al. The Effect of Activated Charcoal on Tissue Cultures:Adsorption of Metabolites Inhibiting Morphogenesis[J]. Physiologia Plantarum,1978,43(2):104-106.
19. Weatherhead M A,Burdon J,Hensgaw G G. Effects of Activated Charcoal as an Additive Plant Tissue Culture Media:Part 2[J]. ZeitschriftFür Pflanzenphysiologie,1979,94(5):399-405.
20. Kohlenbach H W,Wernicke W. Investigations on the Inhibitory Effect of Agar and the Function of Active Carbon in Anther Culture[J]. ZeitschriftFür Pflanzenphysiologie,1978,86(5):463-472.
21. Maene L,Debergh P. Liquid medium additions to established tissue cultures to improve elongation and rooting in vivo[J]. Plant Cell,Tissue and Organ Culture (PCTOC),1985,5(1):23-33.
22. Constantin M J,Henke R R,Mansur M A. Effect of activated charcoal on callus growth and shoot organogenesis in tobacco[J]. In Vitro Cellular&Developmental Biology-Plant,1977,13(5):293-296.
23. Scholl R L,Keathley D E,Baribault T J. Enhancement of Root Formation and Fertility in Shoots Regenerated From Anther and Seedling-Derived Callus Cultures of Arabidopsis thaliana[J]. ZeitschriftFür Pflanzenphysiologie,1981,104(3):225-231.
24. Johansson L,Andersson B,Eriksson T. Improvement of anther culture technique:Activated charcoal bound in agar medium in combination with liquid medium and elevated CO₂ concentration[J]. Physiologia Plantarum,2010,54(1):24-30.
25. Bourgin J P,Nitsch J P. Production of haploids nicotiana from excised stamens[J]. Annales De Physiologie Vegetale,1967,9(4):377-382.
26. Johansson L B,Calleberg E,Gedin A. Correlations between activated charcoal,Fe-EDTA and other organic media ingredients in cultured anthers of Anemone canadensis[J]. Physiologia Plantarum,1990,80(2):243-249.
27. Rogers A. Peonies:5th ed[M]. Oregon:Timber Press,1995:91-105.
28. 李萍,成仿云,张颖星. 防褐剂对牡丹组培褐化发生、组培苗生长和增殖的作用[J]. 北京林业大学学报,2008,30(2):71-76. Li P,Cheng F Y,Zhang Y X. Effects of browning antagonists on antibrowning,growth and multiplication of tissue culture of tree peony[J]. Journal of Beijing Forestry University,2008,30(2):71-76.
29. Pan M J,Van Staden J. The use of charcoal in in vitro culture-A review[J]. Plant growth regulation,1998,26(3):155-163.
30. Kuhne U,Gennerich S. Possibilities of improving in vitro rooting in pome and stone fruits[J]. Gartenbauwissenschaft,1988,35(6):180-182.
31. 卜学贤,陈维伦. 活性碳对培养基中植物生长调节物质的吸附作用[J]. 植物生理学报,1998,14(4):401-405. Bu X X,Chen W L. The effect of activated charcoal on the adsorption of plant regulators in culture medium[J]. ActaPhytophysiologicaSinica,1988,14:401-405.
32. 刘用生,曾兆云,王秀芹等,活性炭吸附作用的生物鉴定[J]. 植物生理学通讯,1995,31(2):123-126. Liu Y S,Zeng Z Y,Wang X Q. The Biological Assay for the Adsorption of Activated Charcoal[J]. Plant Physiology Communications,1995:123-125.
33. Takayama S,Misawa M. Differentiation in Liliumbulbscales grown in vitro. Effects of activated charcoal,physiological age of bulbs and sucrose concentration on differentiation and scale leaf formation in vitro[J]. Physiologia Plantarum,1980,48(1):121-125.
34. 王港,杨秀平,李周岐. 活性炭对组织培养中几种植物激素的吸附作用[J]. 林业科技开发,2006,20(6):26-27. Wang,Yang X P,Li Z Q. Absorption Effect of Activated Charcoal to Hormones during Plant Tissue Culture[J]. China Forestry Science and Technology,2006,20(6):26-27.
35. 何桂梅. 牡丹远缘杂交育种及其胚培养与体细胞胚发生的研究[D]. 北京:北京林业大学,2006. He G M. Studies on Distant Cross-breeding and Embryo in vitro Culture and Somatic Embryogenesis in Tree Peonies[D]. Beijing:Beijing Forestry University,2006.
36. Bouza L,Jacques M,Miginiac E. Requirements for in vitro rooting of Paeonia suffruticosa Andr. cv. ‘Mme de Vatry’[J]. Scientiahorticulturae,1994,58(3):223-233.
37. 曹小勇. 濒危植物紫斑牡丹胚离体培养[J]. 氨基酸和生物资源,2003,25(2):35-36. Cao X Y. In Vitro Culture of Embryo of Endangered Plant Paeonia rockii[J]. Amino Acids and Biotic Resources,2003,25(2):35-36.

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[3]	ZHAO Lian-Fei;CHEN Feng-Li;SUN Xi-Xiao;DU Xin-Qi;ZHAO Chun-Jian*;YAO Huan-Huan;YANG Lei;WANG Gui-Bin;ZU Yuan-Gang. Microwave-assisted Extraction of Paeoniflorin and Paeonol from Tree Peony Seed Coat and Content Determination [J]. Bulletin of Botanical Research, 2014, 34(4): 561-566.
[4]	ZHOU Qi-Chao;YAO Huan-Huan;CHEN Feng-Li;DU Xin-Qi;ZHAO Chun-Jian*;WANG Hua;YANG Lei;WANG Gui-Bin;ZU Yuan-Gang. Optimization of Microwave-assisted Extraction of Paeoniflorin and Paeonol from Peony Pod Using Response Surface Methodology [J]. Bulletin of Botanical Research, 2014, 34(2): 271-275.
[5]	REN Li;YUE Hua*. Cold Resistance of 4 types of Central Plains Tree Peony Cultivars in Harbin Area [J]. Bulletin of Botanical Research, 2010, 30(2): 228-237.