Effects of Rare Earth Element Lanthanum on Adventitious Buds Induction, Plantlets Growth and Synthesis of Secondary Metabolites of Dendrobium officinale

doi:10.7525/j.issn.1673-5102.2020.06.006

Abstract

Abstract:

The experiment was conducted to study the effects of rare earth element lanthanum on adventitious buds induction， plantlets growth and secondary metabolites synthesis of Dendrobium officinale by adding 30-150 μmol·L^-1 La（NO₃）₃ to the media. The effects of 70-90 μmol·L^-1 La³⁺on induction and growth of adventitious buds were the best. The treatment of 130 μmol·L^-1 La³⁺could significantly increase chlorophyll content of seedlings， and 110 μmol·L^-1 La³⁺ could significantly promote plant growth with 15.22 times of fresh weight， and 12.05∶100 of dry weight-fresh weight ratio， which was 64.39% higher than that of control. A certain concentration of La^{3 +} could significantly improve the content of four components in D.officinale. The polysaccharide content reached the peak value of 98.84 mg·g^-1 under 110 μmol·L^-1 La³⁺treatment. The contents of flavonoids， phenolic acid and bibenzyl reached maximum value of 4.31， 7.56 and 21.01 mg·g^-1， respectively， when 90 μmol·L^-1 La³⁺added to media. Both indexes of bibenzyl and flavonoids were highly correlated with other five indexes， while phenolic acid and chlorophyll content were little correlated with other indexes. This study laid the foundation for further exploring the mechanism of rare earth on promoting growth and improving quality of D.officinale， and provided a theoretical basis for scientific and rational utilization of rare earth elements in plant tissue culture.

Key words: Dendrobium officinale Kimura et Migo, lanthanum, culture in vitro, polysaccharides, total flavonoids, total phenols, bibenzyl

CLC Number:

S688.2

Yong-Mei MIAO, Yuan Tong, Da Fang, Yu-Hao Wang. Effects of Rare Earth Element Lanthanum on Adventitious Buds Induction, Plantlets Growth and Synthesis of Secondary Metabolites of Dendrobium officinale[J]. Bulletin of Botanical Research, 2020, 40(6): 839-845.

Figures/Tables 5

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Fig.1

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References 26

1	刘亚娟.铁皮石斛多糖对小鼠胚胎干细胞生长的影响及抗癌免疫活性研究［D］.杭州：浙江工业大学，2014.
	Liu Y J.Effect of Dendrobium officinale polysaccharides on mouse embryonic stem cells and study on its antitumor and immune activity［D］.Hangzhou：Zhejiang University of Technology，2014.
2	陈世锋.铁皮石斛的快速繁殖及其次生代谢产物的研究［D］.武汉：华中农业大学，2006.
	Chen S F.Study on the rapid propagation and secondary metabolites of Dendrobium officinale Kimura et Migo［D］.Wuhan：Huazhong Agricultural University，2006.
3	刘慧雯.LED光质对铁皮石斛组培拟原球茎和幼苗生长及主要有效成分的影响［D］.泰安：山东农业大学，2017.
	Liu H W.Effects of LED light qualities on the growth and main effective ingredients of protocorm-like bodies and seedlings in vitro of Dendrobium officinale［D］.Tai’an：Shandong Agricultural University，2017.
4	张金艳.铁皮石斛两步培养法及有效成分积累的研究［D］.新乡：河南师范大学，2013.
	Zhang J Y.Studies on two-step cultivation and the accumulation of active component of Dendrobium candidum［D］.Xinxiang：Henan Normal University，2013.
5	林弋凯.三种栽培对铁皮石斛C13生长与功效成分的影响［D］.杭州：浙江农林大学，2017.
	Lin Y K.Effects of three cultivation environments on the growth and functional components in Dendrobium catenatum C13［D］.Hangzhou：Zhejiang A&F University，2017.
6	林啸.光质诱导钙信号对铁皮石斛原球茎生长及多糖积累的影响研究［D］.雅安：四川农业大学，2015.
	Lin X.Influence of calcium signal on the growth and polysaccharide accumulation of Dendrobium officinale protocorm under photoinduction［D］.Ya’an：Sichuan Agricultural University，2015.
7	朱波，刘京晶，斯金平，等.铁皮石斛内生真菌对宿主组培苗生长与代谢成分的影响［J］.中国中药杂志，2016，41（9）：1602-1607.
	Zhu B，Liu J J，Si J P，et al.Effects of endophytic fungi from Dendrobium officinale on host growth and components metabolism of tissue culture seedlings［J］.China Journal of Chinese Materia Medica，2016，41（9）：1602-1607.
8	卢敏.石斛种子内生真菌及对石斛苗生长和耐高温的影响［D］.重庆：西南大学，2018.
	Lu M.Endophytes isolated from Dendrobium seeds and their effects on growth and high temperature resistance of Dendrobium seedlings［D］.Chongqing：Southwest University，2018.
9	段晓宇，汪维双，杨红，等.硝酸镧、硝酸铈对细茎石斛组培苗生长的影响［J］.四川农业大学学报，2012，30（2）：186-189.
	Duan X Y，Wang W S，Yang H，et al.Effects of La（NO₃）₃ and Ce（NO₃）₃ on the growth of Dendrobium moniliforme plantlets in vitro［J］.Journal of Sichuan Agricultural University，2012，30（2）：186-189.
10	张桂芳，闫小巧，王艳，等.稀土元素镧、铈对铁皮石斛组培苗生长影响的研究［J］.广东药学院学报，2016，32（5）：559-564.
	Zhang G F，Yan X Q，Wang Y，et al.Effects of rare-earth elements La，Ce on test-tube seedling growth of Dendrobium officinale Kimura et Migo［J］.Journal of Guangdong Pharmaceutical University，2016，32（5）：559-564.
11	徐晓燕，唐迪，蔡菁雯，等.镧对铁皮石斛幼苗生理特性及品质的影响［J］.湖北农业科学，2015，54（12）：2935-2938，3014.
	Xu X Y，Tang D，Cai J W，et al.Effect of La³⁺ on physiological characteristics and quality of Dendrobium officinale seedlings［J］.Hubei Agricultural Sciences，2015，54（12）：2935-2938，3014.
12	李玲，邓晓兰，赵兴兵，等.铁皮石斛化学成分及药理作用研究进展［J］.肿瘤药学，2011，1（2）：90-94.
	Li L，Deng X L，Zhao X B，et al.Advances in studies on chemical constituents in Dendrobium candidum and their pharmacological effects［J］.Anti-Tumor Pharmacy，2011，1（2）：90-94.
13	苗永美，孙佳琦，徐荣华，等.广东石豆兰多糖的提取工艺及其抗氧化活性［J］.天然产物研究与开发，2019，31（5）：779-785.
	Miao Y M，Sun J Q，Xu R H，et al.Optimization of the extraction process of polysaccharide from Bublophyllum kwangtungense and its antioxidant activity［J］.Natural Product Research and Development，2019，31（5）：779-785.
14	苗永美，简兴，汪雁，等.广东石豆兰不同溶剂提取物抗氧化及与总黄酮、总酚含量的关系［J］.核农学报，2020，34（5）：1038-1046.
	Miao Y M，Jian X，Wang Y，et al.Antioxidant activities and their relationship with contents of total flavonoids and total phenols of various solvent extracts from Bublophyllum kwangtungense［J］.Journal of Nuclear Agricultural Sciences，2020，34（5）：1038-1046.
15	贾芳，夏厚林，黄萍，等.紫外可见分光光度法测定叠鞘石斛中联苄类化合物的含量［J］.成都中医药大学学报，2014，37（1）：7-10.
	Jia F，Xia H L，Huang P，et al.Determination of bibenzyl compounds content in Dendrobinm denneanum by UV-visible spectrophotometry［J］.Journal of Chengdu University of Traditional Chinese Medicine，2014，37（1）：7-10.
16	高叶青，任冬梅.稀土元素对短叶对齿藓生理生化的研究［J］.植物研究，2018，38（5）：675-681.
	Gao Y Q，Ren D M.Influence of rare earth elements on physiological and biochemical characteristics of Didymodon tectorum［J］.Bulletin of Botanical Research，2018，38（5）：675-681.
17	王建安，沙莎，李艳芝，等.Ce（NO₃）₃对盾叶薯蓣茎段愈伤组织生长和不定根诱导影响的研究［J］.南京师大学报：自然科学版，2010，33（1）：94-97.
	Wang J A，Sha S，Li Y Z，et al.Effect of Ce（NO₃）₃ on callus growth and adventitious root of stem from Dioscorea zingberensis［J］.Journal of Nanjing Normal University：Natural Science Edition，2010，33（1）：94-97.
18	Ge F，Wang X D，Zhao B，et al.Effects of rare earth elements on the growth of Arnebia euchroma cells and the biosynthesis of shikonin［J］.Plant Growth Regulation，2006，48（3）：283-290.
19	王艳，郭鸽，闫小巧，等.稀土元素镧、铈对霍山石斛试管苗离体生长的影响［J］.中药材，2017，40（3）：528-531.
	Wang Y，Guo G，Yan X Q，et al.Effects of lanthanum and Cerium on in vitro growth of Dendrobium huoshanense［J］.Journal of Chinese Medicinal Materials，2017，40（3）：528-531.
20	De Oliveira C，Ramos S J，Siqueira J O，et al.Bioaccumulation and effects of lanthanum on growth and mitotic index in soybean plants［J］.Ecotoxicology and Environmental Safety，2015，122：136-144．
21	Hu H Q，Wang L H，Li Y L，et al．Insight into mechanism of lanthanum（Ⅲ） induced damage to plant photosynthesis［J］．Ecotoxicology and Environmental Safety，2016，127：43-50．
22	Wu J Y，Wang C G，Mei X G.Stimulation of taxol production and excretion in Taxus spp. cell cultures by rare earth chemical lanthanum［J］.Journal of Biotechnology，2001，85（1）：67-73.
23	杨世海，刘晓峰，果德安.稀土元素镧对掌叶大黄毛状根和非转化根生长及蒽醌产量的影响［J］.中草药，2004，35（10）：1171-1174.
	Yang S H，Liu X F，Guo D A.Effect of rare-earth elements La³⁺ on growth of hairy roots and normal roots of Rheum palmatum and their yield of anthraquinone［J］.Chinese Traditional and Herbal Drugs，2004，35（10）：1171-1174.
24	张儒，张变玲，黄雪梅，等.镧对人参发根中皂苷积累及合成关键酶基因表达的影响［J］.中国稀土学报，2019，37（3）：361-370.
	Zhang R，Zhang B L，Huang X M，et al.Effects of lanthanum on accumulation of ginsenosides and expression of key enzymes genes of ginsenosides biosynthesis in hairy roots of Panax ginseng［J］.Journal of the Chinese Society of Rare Earths，2019，37（3）：361-370.
25	边丽华，邹琳，周冰谦，等.稀土元素镧对丹参活性物质积累及其合成途径中酶基因表达的影响［J］.中国中药杂志，2016，41（23）：4244-4249.
	Bian L H，Zou L，Zhou B Q，et al.Effect of lanthanum on accumulation of active constituent and key enzymes expression of Salvia miltiorrhiza hairy root［J］.China Journal of Chinese Materia Medica，2016，41（23）：4244-4249.
26	罗建平，曹磊，潘利华，等.稀土元素对怀槐悬浮培养细胞异黄酮合成及氧化还原态的影响［J］.应用与环境生物学报，2008，14（3）：362-365.
	Luo J P，Cao L，Pan L H，et al.Effects of rare earth elements on isoflavone biosynthesis and redox status of maackia amurensis cells in suspension cultures［J］.Chinese Journal of Applied & Environmental Biology，2008，14（3）：362-365.

浓度 Concentration （μmol·L^-1）	出芽率 Germination percentage(%)	芽数 No.of buds	鲜重 Fresh weight (g)
0	54.17±5.96d	1.38±0.39b	1.40±0.58b
30	61.11±5.46c	1.56±0.92ab	1.16±0.36b
50	83.33±13.61a	1.75±0.10ab	3.97±1.28a
70	70.83±8.33b	2.83±0.24a	4.16±0.67a
90	66.67±1.24b	2.78±0.51a	3.99±1.00a
110	62.50±10.97c	2.42±0.97ab	3.20±1.71ab
130	62.50±8.46c	2.04±0.31ab	3.02±1.19ab
150	58.33±6.67c	1.96±0.67ab	2.41±0.73ab

浓度 Concentration（μmol·L^-1）	增重倍数 Weight gain multiplier	鲜重 Fresh weight (g)	干重 Dry weight (g)	干鲜比 Dry weight-fresh weight ratio(%)
0	7.33±0.42c	13.77	1.01	7.33
30	8.05±2.02bc	16.11	1.21	7.51
50	9.08±1.14bc	21.08	1.77	8.40
70	9.50±2.25bc	17.76	1.56	8.78
90	10.11±1.97bc	15.35	1.43	9.32
110	15.22±1.45a	16.43	1.98	12.05
130	11.55±1.60b	19.46	1.55	7.97
150	8.52±1.86bc	19.03	1.49	7.83

浓度 Concentration （μmol·L^-1）	多糖含量 Polysaccharide content (mg·g^-1)	总黄酮含量 Total flavonoids content (mg·g^-1)	总酚含量 Total phenols content (mg·g^-1)	联苄含量 Bibenzyl content (mg·g^-1)
0	77.24±0.30e	3.05±0.25d	6.67±0.06ed	12.12±0.12e
30	79.92±0.55d	3.42±0.21cd	6.81±0.17de	12.50±0.12e
50	82.08±0.62c	3.59±0.21bcd	7.15±0.14bc	14.67±0.24d
70	83.20±0.42c	4.12±0.31ab	7.38±0.10ab	17.84±0.12c
90	89.36±0.85b	4.31±0.16a	7.56±0.02a	21.01±0.31a
110	98.84±0.66a	3.83±0.08ab	6.97±0.04cd	19.91±0.08b
130	71.00±1.21f	3.40±0.25cd	6.46±0.17f	19.57±0.19b
150	37.32±1.07g	3.18±0.49cd	5.35±0.10g	12.43±0.19e

	叶绿素含量 chlorophyll content	增重倍数 Weight gain multiplier	干鲜比 Dry weight-fresh weight ratio	多糖含量 Polysaccharide content	总黄酮含量 Total flavonoids content	总酚含量 Total phenols content	联苄含量 Bibenzyl content
叶绿素含量Chlorophyll content	1.000	0.612	0.387	0.043	0.525	0.079	0.779^*
增重倍数Weight gain multiplier	0.612	1.000	0.895^**	0.474	0.404	0.194	0.754^*
干鲜比Dry weight-fresh weight ratio	0.387	0.895^**	1.000	0.593	0.576	0.365	0.671
多糖含量Polysaccharide content	0.043	0.474	0.593	1.000	0.602	0.894^**	0.548
总黄酮含量Total flavonoids content	0.525	0.404	0.576	0.602	1.000	0.761^*	0.769^*
总酚含量Total phenols content	0.079	0.194	0.365	0.894^**	0.761^*	1.000	0.532
联苄含量Bibenzyl content	0.779^*	0.754^*	0.671	0.548	0.769^*	0.532	1.000
平均值Mean	0.404	0.556	0.581	0.526	0.606	0.471	0.676

[1]	Mengqiao GUO, Xuanyu CHEN, Senrong HONG, Jiao LI, Jie FAN, Xinyu CHENG. The Correlation Between Leaf Phenotype Diversity and Total Flavonoids Content of Overground Part of Tetrastigma hemsleyanum Diels & Gilg [J]. Bulletin of Botanical Research, 2022, 42(5): 876-885.
[2]	Tongtong Sun, Zhongjuan Wang, Xiujuan Zhang, Shihan Yang, Xiaoqiang Chen, Ying Zhang. Optimization on Extraction of Total Phenols by Response Surface from Saxifraga stolonifera and Its Correlation in Inhibiting the Activity of PLA₂ in Deinagkistrodon acutus Venom [J]. Bulletin of Botanical Research, 2022, 42(2): 259-267.
[3]	ZHU Ya-Wei, WANG Li-Tao, AN Juan-Yan, Lü Mu-Jie, SUN Jian-Hang, GUO Na, FU Yu-Jie. Response Surface Optimized Enzymatic Hydrolysis-microwave Assisted Extraction Process of Mulberry Leaf Polysaccharide [J]. Bulletin of Botanical Research, 2020, 40(4): 635-640.
[4]	JIANG Shou-Gang, TAN Xiao, WU Lei, WU Bi, YUAN Ying-Jie, ZU Yuan-Gang. Study of the Extraction Technology and Antitumor Activity in vitro of Water-Soluble Polysaccharides from the Wick of Helianthus annuus L. [J]. Bulletin of Botanical Research, 2018, 38(5): 795-800.
[5]	ZHANG Yuan-Yuan, MENG Yong-Bin, XU Lei, JIA Nan, MENG Qing-Huan, XIANG Feng-Ying, LU Qi. Homogenate Extraction Process in Total Flavonoids from Elaeagnus angustifolia Fruit [J]. Bulletin of Botanical Research, 2018, 38(4): 625-631.
[6]	LUO Meng, HU Jiao-Yang, SONG Zhuo-Yue, MU Fan-Song, YU Xue-Ying, QIAO Qi, RUAN Xin, YANG Xuan, ZU Yuan-Gang. Seasonal Variation of Total Flavonoids from Crataegus pinnatifida and Correlation Analysis of Climatic Factors [J]. Bulletin of Botanical Research, 2016, 36(3): 476-480.
[7]	LUO Meng1,2；SONG Zhuo-Yue1,2；HU Jiao-Yang1,2；MU Fan-Song1,2；QIAO Qi1,2；ZU Yuan-Gang1,2. Optimization of Ultrasonic-assisted Extraction of Total Flavonoids from Crataegus pinnatifida* Leaves and Its Antioxidant Activities [J]. Bulletin of Botanical Research, 2015, 35(4): 632-637.
[8]	JIANG Hua-Jun;LIN Yong;CHANG Qiu;WANG Ying-Zi;HUANG Zhi-Qiang;LIU Zhong-Hua. Analysis of the Contents of Total and Three Major Flavonoids in Different Tissues of Mosla chinensis* [J]. Bulletin of Botanical Research, 2013, 33(2): 252-256.
[9]	LIU Jian-Xin;WANG Rui-Juan;JIA Hai-Yan;. Alleviation of SNP on Growth Inhibition and Root Oxidative Damage Caused by Lanthanum in Ryegrass Seedlings [J]. Bulletin of Botanical Research, 2012, 32(6): 680-684.
[10]	WANG Qiu-Xue;GU Cheng-Bo;*;FU Li-Nan;LI Wang;YUAN Xiao-Han;ZU Yuan-Gang;FU Yu-Jie;. Optimization of Cellulase-assisted Extraction Process of Total Flavonoids from Pigeon Pea Leaves Using Response Surface Methodology [J]. Bulletin of Botanical Research, 2012, 32(6): 765-769.
[11]	CHEN Jing;YANG Rui-Lin;ZHU Guang-Long;ZHANG Qiang;WEI Xue-Zhi. Anatomical Structure and Histochemical Localization of the Leaf of Prinsepia uniflora* Batal [J]. Bulletin of Botanical Research, 2012, 32(3): 275-278.
[12]	ZHANG Ying-Cai;YANG Jun;LI Rui. Accumulation and Distribution Characteristics of Rhubarb Polysaccharides in Root of Rheum palmatum* L. [J]. Bulletin of Botanical Research, 2011, 31(6): 744-749.
[13]	ZHAO En-Ze;JIN Shi;WEI Zuo-Fu;WEI Wei;FU Yu-Jie;*. Comparison of the Determination Methods for Total Flavonoids in Pigeon Pea Leaves [J]. Bulletin of Botanical Research, 2011, 31(4): 499-502.
[14]	GUO Cai-Zhen;XUE Dan;HOU Xue-Min;ZHANG Zhi-Feng;YAN Gui-Qin. Optimization of Enzyme-assisted Extraction of Total Flavonoids from Elaeagnus mollis* Diels Leaves by Using Response Surface Methodology [J]. Bulletin of Botanical Research, 2011, 31(3): 371-376.
[15]	SUI Yu-Xiang;ZHANG Jia-Ming;MU Nan;DU Zhen;YAN Gui-Qin. Separation and Purification Process of Total Flavonoids from Elaeagnus mollis* Leaves [J]. Bulletin of Botanical Research, 2010, 30(3): 373-378.