Embryonic Callus Induction and Plant Regeneration of Lilium leucanthum

doi:10.7525/j.issn.1673-5102.2019.03.003

Abstract

Abstract: With Lilium leucanthum Bakeri bulbs as explants, L₉(3⁴) orthogonal test was used to study the effects of different hormones and their concentrations on its callus induction, bulblet differentiation and rooting culture. Histological observation of embryogenic callus was carried out by paraffin sectioning technique. The optimal medium combinations for inducing callus of L.leucanthum under light and dark conditions were MS+TDZ 0.5 mg·L^-1+NAA 1.0 mg·L^-1+2, 4-D 0.05 mg·L^-1 and MS+TDZ 0.5 mg·L^-1+NAA 1.5 mg·L^-1+2, 4-D 0.1 mg·L^-1, with the induction rates of 62.690 and 59.423, respectively. The medium suitable for bulblet rooting was 1/2MS+NAA 0.2 mg·L^-1+1 g AC. After 60-day culture, the seedlings were transferred to peat:vermiculite:perlite=1:1:1 in a medium and cultured in a light incubator for 35 days with the survival rate of 90%. Our experiment successfully established the regeneration system of squamous callus in L.leucanthum, which laid a foundation for the research of tissue culture transgenic seedling and the innovation of germplasm resources.

Key words: Lilium leucanthum, embryonic callus, regeneration system

CLC Number:

Q949.71

ZHANG Lu, PAN Yuan-Zhi, LIU Shi-Liang, CHEN Yuan, SHEN Li-Jia. Embryonic Callus Induction and Plant Regeneration of Lilium leucanthum[J]. Bulletin of Botanical Research, 2019, 39(3): 338-346.

References

1. Mazri M A,Meziani R,Belkoura I,et al. A combined pathway of organogenesis and somatic embryogenesis for an efficient large-scale propagation in date palm(Phoenix dactylifera L.) cv. Mejhoul[J]. Biotech,2018,8(4):215.
2. Heringer A S,Santa-Catarina C,Silveira V. Insights from proteomic studies into plant somatic embryogenesis[J]. Proteomics,2018,18(5-6):1700265.
3. 孙安妮,张延龙,牛立新,等. 宜昌百合体细胞胚诱导及植株再生[J]. 西北农业学报,2011,20(8):142-146. Sun A N,Zhang Y L,Niu L X,et al. Somatic embryogenesis and plantlet regeneration of Lilium leucanthum[J]. Acta Agriculturae Boreali-occidentalis Sinica,2011,20(8):142-146.
4. Nhut D T,Van Le B,Minh N T,et al. Somatic embryogenesis through pseudo-bulblet transverse thin cell layer of Lilium longiflorum[J]. Plant Growth Regulation,2002,37(2):193-198.
5. Tribulato A,Remotti P C,Löffler H J M,et al. Somatic embryogenesis and plant regeneration in Lilium longiflorum Thunb[J]. Plant Cell Reports,1997,17(2):113-118.
6. 陈龙. 百合(Lilium)鳞茎薄层胚性愈伤组织诱导及再生植株遗传稳定性鉴定[D]. 杨凌:西北农林科技大学,2013. Chen L. Direct somatic embryogenesis from bublet thin cell layers of Lilium and assessment of genetic stability in regenerants by ISSR[D]. Yangling:Northwest A&F University,2013.
7. 胡凤荣,席梦利,刘光欣,等. 东方百合的器官发生与体胚发生研究[J]. 南京林业大学学报:自然科学版,2007,31(2):5-8. Hu F R,Xi M L,Liu G X,et al. Study on organogenesis and somatic embryogenesis of oriental lily[J]. Journal of Nanjing Forestry University:Natural Sciences Edition,2007,31(2):5-8.
8. 王杰,刘国锋,包满珠,等. 麝香百合胚性愈伤组织状态的调整与植株再生[J]. 园艺学报,2008,35(12):1795-1802. Wang J,Liu G F,Bao M Z,et al. Adjustment of the status of embryogenic callus and plant regeneration of Lilium longiflorum[J]. Acta Horticulturae Sinica,2008,35(12):1795-1802.
9. 杨柏云,杨慧琴,蔡奇英,等. 龙牙百合体细胞胚的诱导及植株再生[J]. 南昌大学学报:理科版,2005,29(6):536-539. Yang B Y,Yang H Q,Cai Q Y,et al. Somatic embryo induction and plant regeneration of Lilium longii[J]. Journal of Nanchang University:Natural Science,2005,29(6):536-539.
10. 薛冰洋. 两种中国原产百合体细胞胚发生发育的组织学初探[D]. 沈阳:沈阳农业大学,2017. Xue B Y. Histological research of somatic embryogenesis and development in two Lily species native to China[D]. Shenyang:Shenyang Agricultural University,2017.
11. 熊丹,陈发菊,梁宏伟,等. 宜昌百合的组织培养研究[J]. 福建林业科技,2007,34(4):91-94,97. Xiong D,Chen F J,Liang H W,et al. Studies on the tissue culture of Lilium leucanthum[J]. Journal of Fujian Forestry Science and Technology,2007,34(4):91-94,97.
12. 薛晓娜,张延龙,牛立新. 秦岭野生宜昌百合的组织培养研究[J]. 西北农林科技大学学报:自然科学版,2007,35(6):108-112. Xue X N,Zhang Y L,Niu L X. Tissue culture of wild ‘Qinling Yichang’ lily[J]. Journal of Northwest A&F University:Natural Science Edition,2007,35(6):108-112.
13. 刘宝骏,刘国锋. 安祖花胚性愈伤组织诱导及植株再生研究[J]. 热带亚热带植物学报,2018,26(4):407-414. Liu B J,Liu G F. Studies on embryogenic callus induction and plant regeneration of Anthurium andreanum[J]. Journal of Tropical and Subtropical Botany,2018,26(4):407-414.
14. 闫海霞,蒋月喜,邓杰玲,等. 3种东方百合离体培养[J]. 南方农业学报,2016,47(6):889-894. Yan H X,Jiang Y X,Deng J L,et al. In vitro culture for 3 varieties of oriental lily[J]. Journal of Southern Agriculture,2016,47(6):889-894.
15. 刘航空,韩明玉,禹婷,等. 影响油桃叶片产生胚性愈伤组织的因素[J]. 果树学报,2006,23(3):370-374. Liu H K,Han M Y,Yu T,et al. Factors affecting embryonic callus from leaves of early season nectarine cultivars[J]. Journal of Fruit Science,2006,23(3):370-374.
16. 祁宏英,徐洪国,杨伟丽. 用正交试验法优化香水百合试管鳞茎的诱导[J]. 江西农业学报,2018,30(11):32-34. Qi H Y,Xu H G,Yang W L. Optimization of test-tube bulblet inducement of lilium oriental hybrid ‘Casa Blanca’ through orthogonal test[J]. Acta Agriculturae Jiangxi,2018,30(11):32-34.
17. 任建宏,王鹏丽,殷丽娜,等. 平陆百合愈伤组织诱导的正交试验研究[J]. 山西农业大学学报:自然科学报,2017,37(3):189-193. Ren J H,Wang P L,Yin L N,et al. Callus induction of Lilium brownii var. viridulum by orthogonal test[J]. Journal of Shanxi Agricultural University:Natural Science Edition,2017,37(3):189-193.
18. 张艺萍,吴丽芳,吴学尉,等. 东方百合胚性愈伤组织诱导和植株再生研究[J]. 江西农业学报,2008,20(12):33-36,39. Zhang Y P,Wu L F,Wu X W,et al. Embryonic callus inducement and plantlet regeneration of oriental lily[J]. Acta Agriculturae Jiangxi,2008,20(12):33-36,39.
19. Capelle S C,Mok D W S,Kirchner S C,et al. Effects of thidiazuron on cytokinin autonomy and the metabolism of N⁶-(Δ²-Isopentenyl)[8-14C] adenosine in callus tissues of Phaseolus lunatus L. [J]. Plant Physiology,1983,73(3):796-802.
20. 张翔宇,陈杰,吉云,等. 淡黄花百合珠芽诱导愈伤组织再分化出丛生芽的研究[J]. 北方园艺,2015,(19):101-105. Zhang X Y,Chen J,Ji Y,et al. Study on buibil induction callus differentiation from cluster buds of Lilium sulphureum[J]. Northern Horticulture,2015,(19):101-105.
21. Akram M,Aftab F. Establishment of embryogenic cultures and efficient plant regeneration system from explants of forced softwood shoots of teak(Tectona grandis L.)[J]. Horticultural Plant Journal,2016,2(5):293-300.
22. 高洁,王元忠,黄衡宇. 绿花百合胚性愈伤组织诱导与植株再生研究[J]. 植物研究,2016,36(1):52-57. Gao J,Wang Y Z,Huang H Y. Embryonic callus induction and plant regeneration of Lilium fargessi[J]. Bulletin of Botanical Research,2016,36(1):52-57.
23. 狄翠霞,安黎哲,张满效,等. 西伯利亚百合器官离体培养及结鳞茎的研究[J]. 西北植物学报,2005,25(10):1931-1936. Di C X,An L Z,Zhang M X,et al. In vitro culture and bulblet formations of Lilium×siberia Explants[J]. Acta Botanica Boreali-Occidentalia Sinica,2005,25(10):1931-1936.
24. 林娅,郑玉梅,刘青林. 影响月季愈伤组织诱导和分化的因素[J]. 分子植物育种,2006,4(2):223-227. Lin Y,Zheng Y M,Liu Q L. Factors in callus induction and differentiation on Rosa[J]. Molecular Plant Breeding,2006,4(2):223-227.
25. 高永超,王加宁,邱维忠,等. 不同光照和植物生长物质对地钱愈伤组织诱导及分化的影响[J]. 山东科学,2007,20(4):37-39,43. Gao Y C,Wang J N,Qiu W Z,et al. Effects of different light and plant growth substances on the induction and decomposition of callus of Marchantia polymorpha[J]. Shandong Science,2007,20(4):37-39,43.
26. 李伟. 西葫芦未受精子房离体培养成胚条件优化与成苗技术研究[D]. 杨凌:西北农林科技大学,2012. Li W. Study on the optimization of embryoids culture condition and plantlet formation in summer squash in vitro ovary culture techniques[D]. Yangling:Northwest A&F University,2012.
27. 魏海霞,刘德玺,王霞,等. 2苹果品种叶片胚状体再生体系的建立[J]. 西北林学院学报,2014,29(1):84-88,94. Wei H X,Liu D X,Wang X,et al. Establishment of regeneration system of embryoid from the leaves of two apple varieties[J]. Journal of Northwest Forestry University,2014,29(1):84-88,94.
28. 孔凡芹,李树忠,黄玮婷,等. 楤木胚状体的诱导及再生体系的建立[J]. 江苏农业科学,2018,46(7):36-39. Kong F Q,Li S Z,Huang W T,et al. Induction of embryos and establishment of regeneration system of Aralia chinensis[J]. Jiangsu Agricultural Science,2018,46(7):36-39.
29. 严瑞. 基于体细胞胚发生的五种切花百合高效再生体系建立[D]. 沈阳:沈阳农业大学,2017. Yan R. Establishment efficient regeneration system of five cut-flower varieties Lilium based on somatic embryogenesis[D]. Shenyang:Shenyang Agricultural University,2017.
30. 黄哲,向景葵. 内源激素对黄姜体细胞胚发生的影响[J]. 中国农学通报,2007,23(2):255-258. Huang Z,Xiang J K. A study on endogenous phytohormones changes during somatic embryogenesis in dioscorea zingiberensis wright[J]. Chinese Agricultural Science Bulletin,2007,23(2):255-258.
31. 鲁娇娇,严瑞,何香杉,等. 朱顶红‘Red Lion’胚性愈伤组织诱导及体细胞胚发生[J]. 园艺学报,2016,43(12):2451-2460. Lu J J,Yan R,He X S,et al. The embryonic callus induction and somatic embryogenesis of Hippeastrum vittatum ‘Red Lion’[J]. Acta Horticulturae Sinica,2016,43(12):2451-2460.
32. 肖望,涂红艳,张爱玲. 白姜花胚性愈伤组织的诱导与植株再生体系的建立[J]. 园艺学报,2016,43(8):1605-1612. Xiao W,Tu H Y,Zhang A L. Embryogenic callus induction and plant regeneration from Hedychium coronarium via Somatic Embryogenesis[J]. Acta Horticulturae Sinica,2016,43(8):1605-1612.
33. 曹雨,陈章辉,区丁文,等. 墨西哥玉米草组培体系的建立及愈伤组织切片观察[J]. 华南师范大学学报:自然科学版,2016,48(6):19-24. Cao Y,Chen Z H,Qu D W,et al. The establishment of teosinte tissue culture and callus section observation[J]. Journal of South China Normal University:Natural Science Edition,2016,48(6):19-24.
34. 袁雪,钟雄辉,李晓昕,等. 铁炮百合的胚性愈伤组织诱导和植株再生[J]. 核农学报,2012,26(3):454-460. Yuan X,Zhong X H,Li X X,et al. Embryonic callus induction and plant regeneration of Lilium longiflorum[J]. Journal of Nuclear Agricultural Sciences,2012,26(3):454-460.

[1]	ZHANG Jing, NIU Zhe, FAN Wei-Fang, GAO Peng-Fei, WU Jian-Hui. Establishing Tissue Culture and Regeneration System of Scutellaria regeliana [J]. Bulletin of Botanical Research, 2020, 40(1): 50-57.
[2]	GAO Jie1；WANG Yuan-Zhong2；HUANG Heng-Yu1. Embryonic Callus Induction and Plant Regeneration of Lilium fargesii* [J]. Bulletin of Botanical Research, 2016, 36(1): 52-57.
[3]	DU Zhao-Wei；ZHENG Tang-Chun；LI Shuang；ZANG Li-Na；QU Guan-Zheng；YOU Xiang-Ling. Rapid Propagation and Regeneration System of Populus simonii×Populus nigra* [J]. Bulletin of Botanical Research, 2015, 35(6): 904-907.
[4]	LI Fang1；XIAO Xiao-Jun1；LIN Zhong-Quan2；DU Ming-Lin1；HU Zhang-Wei1；HUANG Zuo-Xi1. Establishment and Optimization of Wild Beltilla striata* Regeneration System [J]. Bulletin of Botanical Research, 2015, 35(6): 825-831.
[5]	ZHANG Jian-Ying；QI Yong-Hui；Lü Yue-Dong；LIU Jian-Ming；XING Ya-Juan. Regeneration System for Axillary Bud of Juglans mandshurica* Maxim. [J]. Bulletin of Botanical Research, 2015, 35(1): 22-26.
[6]	HONG Sen-Rong;YIN Ming-Hua;WANG Ai-Ping. Establishment of Callus Induction and Regeneration System of Jiangxi Yanshan Red Bud Taro (Colocasia esculenta* L. Schott var. cormosus CV.Hongyayu) Virus-Free Plantlet Corms [J]. Bulletin of Botanical Research, 2013, 33(6): 738-745.
[7]	YIN Ming-Hua;HONG Sen-Rong. Establishment of Regeneration System of Leaves and Stems of Dioscorea bulbifera* L. Virus-free Plantlets [J]. Bulletin of Botanical Research, 2009, 29(4): 492-499.
[8]	ZU Yuan-gang, KONG Jin, WU Shuang-xiu, NIE Ming-zhu. THE ESTABLISHMENT OF HYPOCOTYL REGENETATION SYSTEM OF BETULA PLATYPHYLLA [J]. Bulletin of Botanical Research, 2001, 21(4): 615-619.
[9]	ZHANG Dong-xiang, ZHANG Chong-hao, MEI Ling, LI Jie-fen. INDUCTION OF EMBRYONIC CALLUS AND FORMATION OF EMBRYOID RELATED TO ENDOGENOUS IAA AND ABA IN CELERY [J]. Bulletin of Botanical Research, 2001, 21(1): 70-73.