Tissue Culture via Seed Embryo for Michelia macclurei

doi:10.7525/j.issn.1673-5102.2021.01.011

Abstract

Abstract:

In order to explore tissue culture and micro-propagation technical system of Michelia macclurei， the seeds from its elite families were used to carry out experimental studies at aspects of explants selection and disinfection， basic medium， PVP concentration， type and concentration of rooting promoter. The results showed that：①The embryos were well sterilized with 0.1% mercury solution for 6-9 min， and the aseptic germination rate could reach 60%-65%； ②LY medium was the best basic medium， and suitable for the proliferation and growth of most embryo lines； ③Application of PVP with concentration of 4-8 g·L^-1 could significantly reduce browning degree of medium and promote growth of the shoots； ④There were significant differences in rooting ability and rooting rate among tested embryo lines， the optimal concentration of both rooting promoters was 8.5 mg·L^-1， and the rooting induction effect of ABT2 root powder was significantly better than that of IBA（P<0.01）； ⑤The survival rate reached above 90% for rooting plantlets in 30 d， and more than 80% for plantlets with no root in 60 d after transplanted. This study provides a theoretical and technical basis for study on tissue culture and micro-propagation of M.macclurei clones and their industrial production.

Key words: Michelia macclurei Dandy, embryo clone, PVP, basic culture media, browning

CLC Number:

S722.3+7

Ying LIU. Tissue Culture via Seed Embryo for Michelia macclurei[J]. Bulletin of Botanical Research, 2021, 41(1): 79-88.

Figures/Tables 8

Table 1

Fig.1

Table 2

Table 3

Table 4

Table 5

Table 6

Table 7

References 24

1	李振问.火力楠林带的防火效果研究［J］.厦门大学学报：自然科学版，2002，41（5）：596-599.
	Li Z W.Studies on the effect of fire prevention of biological network of Michelia macclurei［J］.Journal of Xiamen University：Natural Science，2002，41（5）：596-599.
2	李清莹，陈伟俊，仲崇禄，等.火力楠种质资源早期综合评价与选择［J］.浙江理工大学学报，2019，41-42（6）：799-805.
	Li Q Y，Chen W J，Zhong C L，et al.Early comprehensive evaluation and selection of Michelia macclurei germplasm resources［J］.Journal of Zhejiang Sci-Tech University，2019，41-42（6）：799-805.
3	韦中绵，覃德文，吴敏，等.不同光照强度对火力楠幼苗生长及生理特性的影响［J］.西部林业科学，2018，47（2）：48-53.
	Wei Z M，Qin D W，Wu M，et al.Growth and physiological characteristics of Michelia macclurei in different light intensities［J］.Journal of West China Forestry Science，2018，47（2）：48-53.
4	姜清彬，李清莹，王里，等.四种方法对火力楠叶片油的提取分析［J］.分子植物育种，2018，16（6）：1985-2000.
	Jiang Q B，Li Q Y，Wang L，et al.Extraction of essential oil from leaves of Michelia macclurei dandy by four extraction methods［J］.Molecular Plant Breeding，2018，16（6）：1985-2000.
5	高柱，王小玲，唐军荣，等.我国木兰科植物组培快繁研究进展［J］.江西科学，2013，31（1）：53-57.
	Gao Z，Wang X L，Tang J R，et al.Advances in tissue culture of Magnoliaceaein China［J］.Jiangxi Science，2013，31（1）：53-57.
6	黄烈健，王鸿.林木植物组织培养及存在问题的研究进展［J］.林业科学研究，2016，29（3）：464-470.
	Huang L J，Wang H.Advances in tissue culture techniques of trees and the problems existed［J］.Forest Research，2016，29（3）：464-470.
7	黄燕芳，周国兰，赵华富.降低茶树组织培养中外植体褐化程度的研究［J］.西南农业学报，2009，22（5）：1492-1495.
	Huang Y F，Zhou G L，Zhao H F.Effect of different culture conditions on browning rate of explants in tissue culture of tea trees［J］.Southwest China Journal of Agricultural Sciences，2009，22（5）：1492-1495.
8	Fett-Neto A G，Dicosmo F，Reynolds W F，et al.Cell culture of Taxus as a source of the antineoplastic drug taxol and related taxanes［J］.Biotechnology，1992，10（12）：1572-1575.
9	柳蔓琼，梁士现.火力楠茎段的组织培养［J］.植物生理学通讯，1985，（1）：37.
	Liu M Q，Liang S X.Tissue culture by shoot segments for Michelia macclurei［J］.Plant Physiology Communications，1985，（1）：37.
10	李雪，王淑芬，蒋雄辉.醉香含笑的组织培养与植株再生［J］.植物生理学通讯，2005，41（6）：783.
	Li X，Wang S F，Jiang X H.Tissue culture and plantlet regeneration of Michelia macclurei Dandy［J］.Plant Physiology Communications，2005，41（6）：783.
11	侯金艳，李明浩，毛颖基，等.黄连木种子组织培养不同消毒方法的研究［J］.中国农学通讯，2013，29（10）：7-10.
	Hou J Y，Li M H，Mao Y J，et al.Effects of different sterile technique on seeds culture in Pistacia chinensis Bunge［J］.Chinese Agricultural Science Bulletin，2013，29（10）：7-10.
12	周芳，徐剑秋，安娟艳，等.短瓣金莲花种子无菌萌发与幼苗快繁技术初探［J］.植物研究，2018，38（4）：619-624.
	Zhou F，Xu J Q，An J Y，et al.Tissue culture and rapid propagation of Trollius ledebouri［J］.Bulletin of Botanical Research，2018，38（4）：619-624.
13	曾炳山，刘英，许煌灿，等.长嘴黄藤离体快繁研究［J］.福建林学院学报，2002，22（2）：169-171.
	Zeng B S，Liu Y，Xu H C，et al.Tissue culture of Daemonorops jenkinsiana［J］.Journal of Fujian College of Forestry，2002，22（2）：169-171.
14	陈菲，李黎，宫伟.植物组织培养的防褐化探讨［J］.北方园艺，2005，（2）：69.
	Chen F，Li L，Gong W.Study on the browning problen in plant tissue culture［J］.Northern Horticulture，2005，（2）：69.
15	孟雪.白玉兰的组织培养和快速繁殖［J］.植物生理学通讯，2005，41（3）：339.
	Meng X.Tissue culture and rapid propagation of Magnolia denudata［J］.Plant Physiology Communications，2005，41（3）：339.
16	陈金慧，施季森，诸葛强.杂交鹅掌楸的不定芽诱导及植株再生［J］.植物生理学通讯，2002，38（5）：459.
	Chen J H，Shi J S，Zhu G Q.Adventitious bud induction and plantlet regeneration of Liriodendron hybrid （L.chinense×L.tulipifera）［J］.Plant Physiology Communications，2002，38（5）：459.
17	高红兵，杜凤国，王欢.抗褐化剂对天女木兰芽外植体褐化与酚酸氧化的影响［J］.林业科学研究，2017，30（3）：525-532.
	Gao H B，Du F G，Wang H.Effects of browning inhibitors on bud explants browning and phenolic acids oxidation of Magnolia sieboldii K.Koch［J］.Forest Research，2017，30（3）：525-532.
18	王倩颖，唐佳妮，刘志高，等.景宁木兰组织培养外植体选择与抗褐化研究［J］.广西植物，2017，37（9）：1088-1095.
	Wang Q Y，Tang J N，Liu Z G，et al.Explant selecting and anti-browning of Magnolia sinostellata in tissue culture［J］.Guihaia，2017，37（9）：1088-1095.
19	Babbar S B，Gupta S C.Induction of androgenesis and callus formation in vitro cultured anthers of a myrtaceous fruit tree （Psidium guajiava L.）［J］.Journal of Plant Research，1986，99（1）：75-83.
20	刘玉壶.中国木兰［M］.北京：北京科学技术出版社，2004.
	Liu Y H.Magnolias of China［M］.Beijing：Peking Science and Technology Press，2004.
21	李艳，王青，李洪艳，等.3种玉兰的组织培养［J］.植物生理学通讯，2005，41（5）：633.
	Li Y，Wang Q，Li H Y，et al.Tissue culture of 3 species of Magnolia L.［J］.Plant Physiology Communications，2005，41（5）：633.
22	Fukaki H，Okushima Y，Tasaka M.Regulation of lateral root formation by auxin signaling in Arabidopsis［J］.Plant Biotechnology，2005，22（5）：393-399.
23	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 S L，Wang H M，Zhang D.Decreasing the hyperhydricity and rooting of Scutellariae baicalensis Georgi［J］.Bulletin of Botanical Research，2016，36（1）：90-96.
24	王媛媛，侯潇棐，李永霞，等.ABT 2号生根粉对西藏不同地区银白杨扦插生根及生长的影响［J］.种子，2019，38（3）：73-76.
	Wang Y Y，Hou X F，Li Y X，et al.Effects of ABT 2 on rooting and growth of Populus alba cuttings in different areas of Tibet［J］.Seed，2019，38（3）：73-76.

消毒时间 Disinfection time(min)	染菌率 Contamination rate(%)	胚死率 Mortality rate(%)	无菌胚成活率 Aseptic retention rate(%)	萌芽率 Germination rate(%)
3	90d	0a	10b	10b
6	35c	0a	65a	65a
9	20bc	17b	63a	60a
12	15b	65c	20b	17b
15	0a	100d	0c	0c

指标 Indicator	处理 Treatment	Ａ１	A２	A3	A４	A５	A６	A７	A８	Ｂ１	Ｂ2
平均芽数(个) Mean number of shoots	MS	2.01abD	1.30bE	2.70bB	1.30aE	2.87bB	2.80bB	2.85bB	2.34bC	2.69 bB	3.02bA
	3/4MS	2.00abD	1.35abE	2.80abB	1.28bcE	3.00abB	2.90abB	2.87abB	2.50abC	2.80abB	3.50abA
	1/2MS	1.67bD	1.30bE	2.50bB	1.11cE	2.60bB	2.50 bB	2.55 bB	2. 00 bC	2.40 bB	3.40bA
	1/4MS	1.50cD	1.10cE	2.00cB	1.00cE	2.30cB	2.23 cB	2.10 cB	1.67cC	2.05cB	2.50cA
	LY	2.71aD	1.56aE	3.31aB	1.28bE	3.30aB	3.30 aB	3.30 aB	2.80 aC	2.92aB	3.50aA
平均苗高 Mean seedling height （cm）	MS	0.70dC	0.50dD	0.98dB	0.80dB	0.78dC	1.20dB	0.78dC	0.60dD	1.23dA	1.18dAB
	3/4MS	0.90cC	0.61cCD	1.17cB	1.15cB	0.86cC	1.26cB	0.96 cC	0.60cD	1.30cA	1.27cAB
	1/2MS	1.20bBC	0.83bD	1.26bAB	1.26bB	1.09bC	1.38bA	1.00bCD	0.78bD	1.35bA	1.37bA
	1/4MS	1.25abC	1.20abD	1.38abB	1.33abBC	1.18abE	1.43abAB	1.18abE	0.86abF	1.54abA	1.44abAB
	LY	1.41aC	1.26aD	1.50aB	1.43aB	1.26Ea	1.57aAB	1.21 aE	0.90aF	1.71aA	1.51aAB
≥3 cm苗占比 Percentage of plantlets ≥3 cm high（%）	MS	0	0	0	0	0	0	0	0	0	0
	3/4MS	0	0	0	0	0	0	0	0	0	0
	1/2MS	0	0	0	0	0	5.3cA	0	0	9.5cA	5.6cA
	1/4MS	5.1bA	0	15.6bA	10.8bA	0	15.7bA	0	0	15.7bA	20.7bA
	LY	30.3aAB	20.5aB	50.7aAB	30.6aAB	15.2aB	40.6aAB	20.3aB	0	50.8aA	30.8aAB

处理 Treatment	A1	A2	A3	A4	A5	A6	A7	A8	B1	B2
MS	5.60aA	5.58aA	5.30aB	5.80aA	5.75aA	3.75aB	5.90aA	5.68aA	3.68aB	5.40aB
3/4MS	4.50bA	4.80bA	4.20bAB	4.90bA	4.80bA	3.59bB	4.50bA	4.89bA	3.30bB	4.78bA
1/2 MS	4.00cA	4.25cA	2.66cB	4.21cA	4.00cA	3.23cAB	3.45cA	4.30cA	2.78cB	3.56cA
1/4MS	2.06dAB	2.80dA	1.86dB	2.38dA	2.78dA	1.98dB	3.32dA	3.20dA	1.96dB	2.40dA
LY	1.06eAB	1.56eAB	0.95eB	1.56eA	1.56eA	0.34eB	2.33eA	2.25eA	1.02eB	1.46eAB

浓度 Concentration (g·L^-1)	A1	A2	A3	A4	A5	A6	A7	A8	B1	B2
0	3.68aA	3.80aA	3.30aAB	3.56aA	3.88aA	1.90aB	3.55aA	3.56aA	2.68bB	3.40bAB
2	2.89bAB	3.35bA	2.87bAB	2.20bB	3.38bA	1.80bB	3.57aA	3.35bA	2.80aB	3.56aA
4	1.26cAB	1.56cAB	1.05cAB	1.56cAB	1.60cA	0.77cB	2.33bA	1.75cA	0.85cB	1.02cAB
6	1.28cA	1.55cA	1.08cAB	1.57cA	1.58cA	0.86cB	1.23cA	1.80cA	0.90cB	0.98cAB
8	1.23cA	1.67cA	1.01cAB	1.50cA	1.62cA	0.83cB	1.16cA	1.78cA	0.88cB	1.16cAB

指标 Traits	浓度 Concentration (g·L^-1)	A1	A2	A3	A4	A5	A6	A7	A8	B1	B2
平均芽数(个) Mean number of shoots	0	2.68aC	1.56aD	3.30 aB	1.30aD	3.40aAB	3.30 aB	3.30 aB	2.80aC	2.92aBC	3.60aA
	2	2.70aC	1.65aD	3.28aAB	1.35aD	3.30aAB	3.29aAB	3.25 aB	2.88aC	2.96aBC	3.55aA
	4	3.01aC	1.70aD	3.30aAB	1.65aD	3.35aAB	3.50 aA	3.23aAB	3.00aC	3.23 aB	3.48aA
	6	3.15aC	1.66aD	3.29aAB	1.60aD	3.29aAB	3.34aAB	3.27 aB	3.12aC	3.05 aC	3.49aA
	8	3.00aC	1.67aD	3.30aAB	1.58aD	3.27 aB	3.30aAB	3.29 aB	3.20aC	3.02 aC	3.50aA
平均苗高 Mean plantlet height (cm)	0	1.43bBC	1.26bC	1.50bB	1.43bBC	1.26bC	1.57bB	1.21bC	0.90bD	1.71bA	1.51bBC
	2	1.45bBC	1.28bC	1.55bBC	1.45bBC	1.35bC	1.60bB	1.25bC	1.15bD	1.73abA	1.56bB
	4	1.58aBC	1.49aC	1.70abBC	1.55aBC	1.50abC	1.75aB	1.47abC	1.20bD	2.07aA	1.87aB
	6	1.60aBC	1.47aC	1.75aB	1.58aBC	1.53aC	1.73aB	1.50aC	1.21aD	2.04aA	1.85aB
	8	1.57aC	1.50aC	1.73aBC	1.53aC	1.58aC	1.75aB	1.49aC	1.31aD	2.00aA	1.89aB
≥3 cm苗占比 Percentage of plantlets ≥3 cm high(%)	0	30.6bB	20.4bD	50.3bA	30.4bB	15.3bD	40.3bA	20.6bD	0	50.3bA	34.0bC
	2	30.4bD	22.3bE	53.6bA	33.5bB	20.4bE	40.7bA	23.5bE	0	52.4bA	35.6bC
	4	50.9aB	35.8aB	70.7aA	50.7aB	51.5aB	75.6aA	43.3aB	0	75.3aA	72.5aA
	6	50.7aCD	33.7aD	70.8aA	50.8aC	49.9aCD	73.5aAB	40.4aD	0	76.2aA	73.1aB
	8	50.8aCD	34.4aD	70.6aA	52.6aC	50.6aCD	75.1aAB	41.2aD	0	75.8aA	72.4aB