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1.
Cinchona officinalis (Rubiaceae) is an endemic species of the Loja Valley in southern Ecuador with medicinal uses. Because of over-exploitation in the nineteenth century and more recent disturbances to its ecosystem, C. officinalis populations are threatened. Currently, natural regeneration of the populations is low, despite its high plant regeneration and seed formation capacity. In the present study, an efficient protocol for germination, shoot proliferation and plantlets regeneration was developed for this species. Phenolic content and germination rate of C. officinalis seeds were compared with a control species, C. pubescens. Nodal segments from seedlings of C. officinalis were cultured on Gamborg medium supplemented with different combinations of plant growth regulators. Because the phenol content is high in C. officinalis, the phenolic should be removed with hydrogen peroxide or water washes to stimulate germination. Shoots and callus developed from nodal segments within 45 days using most of the tested combinations of plant growth regulators. The best rates of shoot proliferation, callus formation and adventitious buds were obtained in medium supplemented with 5.0 mg L?1 6-benzyl-aminopurine and 3.0 mg L?1 indole-3-butyric acid. 相似文献
2.
《林业研究》2019,(6)
This research reports on an efficient shoot proliferation and callus regeneration system for bamboo.Young, semi-lignified branches with one lateral bud from Drepanostachyum luodianense(Yi et R. S. Wang) Keng f.were used as explants. Disinfection with 0.1% HgCl_2 for 8 min was the optimum treatment and the best medium for bud initiation was Murashige and Skoog(MS) medium containing 3.0 mg L~(-1)6-benzyladenine(BA). Multiple shoots were induced from nodal shoot segments on MS medium containing 5.0 mg L~(-1) BA, 0.5 mg L~(-1) kinetin(Kin), and 1.0 mg L~(-1) naphthaleneacetic acid(NAA). The highest frequency of callus formation(65.6%) was on MS medium containing 4.0 mg L~(-1)2,4-dichlorophenoxyacetic acid(2, 4-D), 0.5 mg L~(-1) NAA, and 0.1 mg L~(-1) thidiazuron(TDZ). The optimum medium for callus proliferation was MS medium with 4 mg L~(-1)2,4-D, 0.5 mg L~(-1) TDZ and 0.5 mg L~(-1) NAA, and the optimum hormone combination was 4 mg L~(-1) BA ? 0.5 mg L~(-1) NAA for callus redifferentiation(up to 85.6%). A 100% rooting was achieved on MS medium supplemented with 2.0 mg L~(-1) NAA and 0.5 mg L~(-1)3-indole butyric acid(IBA). Rooted plantlets were acclimatized in a greenhouse in humus soil ? perlite(1:1) substrate. These micropropagated callus induction and regeneration systems for bamboo will be useful for genetic engineering and multiplication. 相似文献
3.
《林业研究》2021,32(2)
The induction and proliferation of embryogenic callus are key steps for large-scale propagation of somatic embryogenesis pathway and long-term preservation of coniferous germplasm.Callus can be induced from immature embryos of Korean pine(Pinus koraiensis Sieb.et Zucc.;Pinaceae) as explants,but there are problems,such as low proliferation efficiency,loss of embryogenicity,poor vigor;thus,best conditions for proliferation and culture of immature embryos of Korean pine are not yet clear.To solve the problems with somatic embryogenesis of Korean pine and determine the best culture conditions for callus induction and proliferation,we varied hormone concentration,subculture cycle of proliferation and other plant growth regulators combinations in media to induce callus formation by megagametophytes of three Korean pine families at different developmental stages,then analyzed the effects on embryogenic callus retention and cell proliferation using a quadratic regression orthogonal rotation design.The results showed that the family origin and collection date of explants significantly affected callus induction(induction rate reached1.67%).Embryogenic maintenance and callus proliferation were best on DCR medium supplemented with 0.25 mg L~(-1)6-benzyl adenine,1 mg L~(-1) naphthaleneacetic acid,30 g L~(-1)sucrose,500 mg L~(-1),L-glutamine,500 mg L~(-1) casein hydrolysis and 6.5 g L~(-1) agar.In addition,the combination of 2,4-dichlorophenoxyacetic acid+6-benzyl adenine also had a better proliferative effect on callus.The effects of different combinations of growth regulators on callus proliferation efficiency were significantly different.Transfer to new medium every 13-15 days not only maintained robust callus vigor,but also yielded a larger proliferation coefficient.The techniques and conditions for embryogenic callus induction and proliferation of Korean determined here will serve as a foundation for establishing a large-scale system for somatic embryogenesis and propagation of Korean pine. 相似文献
4.
柳桉组培快繁技术 总被引：1，自引：0，他引：1
以3年生柳桉优树环割促萌枝条为外植体,以MS为基本培养基,通过激素种类与浓度、大量元素配比等试验,探讨了柳桉腋芽诱导、继代增殖、生根培养基配方。结果表明:外植体宜采用中等幼嫩的茎段,用75%酒精消毒30 s、0.1%升汞溶液消毒4 min效果最佳;在3/4 MS+0.3 mg.L-16-BA+0.2 mg.L-1NAA培养基上进行侧芽诱导,柳桉外植体出芽率最高;在改良MS+0.2 mg.L-16-BA+0.2 mg.L-1NAA继代培养基上,柳桉增殖效果最好,其芽体健壮,生长正常,增殖率3倍以上;应用1/2MS培养基附加ABT 1#0.5 mg.L-1和IBA 0.1 mg.L-1,其生根率可达95%以上,生根3~4条,苗木生长健壮,移栽成活率可达92%以上。这些配方已成功地应用于柳桉优良无性系苗木的工厂化生产,单个超净台月苗木生产能力达7万株以上。 相似文献
5.
Juniperus thurifera L.is an endemic Cupres saceae from the Aure`s Mountains of north eastern Algeria and endangered,in part,due to the scarcity of viable seeds It is threatened by other abiotic factors and the lack of an effective management strategy will increase its risk o extinction.The dearth of information on its in vitro regeneration impedes its application in forest managemen programs.We therefore developed a micropropagation protocol using microcuttings with auxiliary buds.Cuttings were grown on different combinations of media supplemented with plant growth regulators at different concentrations.The highest number of shoots and branches regenerated from original shoots was obtained on Woody Plant Medium(WPM)supplemented with 6-benzylaminopurine(BAP)(0.5 mg L-1)and 2,4-dichlorophe noxyacetic acid(2,4-D)(0.25 mg L-1).The best elongation of shoots was achieved with WPM supplemented with0.5 mg L-1of BAP and 0.25 or 1 mg L-1 of 2,4-D.On the second subculture,shoots had a higher number of branches than those of the first.The highest rooting rate,38.8%,was obtained with shoots cultured in 1/2 Murashige and Skoog(MS)medium supplemented with 5.0 mg L-1each of indol-3-butyric(IBA)and naphthalene acetic acid(NAA).Similarly,the highest root numbers and lengths were produced on 1/2 MS medium supplemented with IBA and NAA(5.0 mg L-1each).During transfer to acclimatization,rates of plant losses of 50% occurred.The second part of the experiment showed that the best shoot callusing was on WPM supplemented with BAP and 2,4-D,with either the combination 0.5+0.25 or 0.25+0.25 mg L-1.The results of this research provide a starting point for further studies on in vitro regeneration of J.thurifera for the sustainable management of its unique ecosystem in the Mediterranean basin. 相似文献
6.
以松塔景天幼嫩叶片作为外植体,利用含有不同浓度配比植物生长调节剂的培养基分别进行愈伤组织的诱导、不定芽的诱导、生根培养及驯化移栽,建立了松塔景天组织培养和快速繁殖体系。试验结果表明：诱导愈伤组织的最适培养基为MS＋6-BA 0．5 mg/L ＋NAA 0．05mg/L ,愈伤组织诱导率达87．78％;诱导不定芽的最适培养基为MS＋6-BA 1．0mg/L＋ NAA0．1 mg/L ,不定芽诱导率达88．89％;组培苗最佳生根培养基为1/2 M S＋IB A 0．3 m g/L ,生根率达91．33％;组培苗的移栽成活率达84％。 相似文献
7.
爬行卫矛下胚轴高频离体再生体系的建立(英文) 总被引：1，自引：0，他引：1
In this paper,a protocol for efficient shoot regeneration was successfully developed from hypocotyl explants of Euonymus fortunei var.radicans.Some factors that influenced shoot regeneration such as different combinations of plant growth regulators,types of medium and inoculation ways were studied in order to establish an efficient plant regeneration for transformation.The results showed that hypocotyl explants wero horizontally cultured on a basic medium composed of MS medium supplemented with 0.5 mg·L-1 BAP and 0.01 mg·L-1 NAA for induction and development of adventidous shoots.Ninety-four percent of regeneration frequency and 5.1 shoots per explants were obtmned after 30 days of culture.Regenerated shootsproliferated efficiently on a shoot multiplication medium consisting of MS medium containing 1.0 mg·L-1 BAP and 0.1 mg·L-1 NAA.Microshoots were rooted on a rooting medium made up of MS medium enriched with O.5 mg·L-1 IBA and O.5 mg·L-1IAA.After hardening,90% of plants were successfully established under greenhouse conditions.Histological observation revealed that shoot primordium originated from subepidermal cells of hypocotyl explants and directly developed into adventitious shoots without caHus formation. 相似文献
8.
9.
Rui Fang Wang Feng Lan Huang Jian Zhang Qiu Yan Zhang Li Na Sun Xing Shun Song 《Journal of Forest Research》2016,21(5):244-250
Cerasus humilis is a species of small, perennial, drought-resistant and multipurpose deciduous shrub grown in arid and semi-arid conditions in northern China. In this study, an efficient protocol for the rapid micropropagation of C. humilis has been standardized using stem and/or leaf explants. Direct multiple shoot induction was observed when the stem explants were cultured on Murashige and Skoog (MS) medium supplemented with different plant growth regulators. The highest shoot induction was obtained when stem explants from adult trees were cultured on MS medium supplemented with 2.0 mg L?1 6-benzyladenine (6-BA) and 0.9 mg L?1 α-naphthaleneacetic acid (NAA). The leaf and stem explants cultured on MS medium with 1.0 mg L?1 6-BA and 0.6 mg L?1 NAA, and 0.5 mg L?1 6-BA and 0.8 mg L?1 NAA, respectively, produced the highest induction frequency of callus. Maximum proliferation of callus was observed on MS medium containing a combination of 0.5 mg L?1 6-BA with 0.6 mg L?1 2,4-dichlorophenoxyacetic acid (2,4-d). Optimal shoots differentiated from callus were obtained on MS medium supplemented with 5.0mg L?1 6-BA and 0.9 mg L?1 NAA. In vitro rooting was achieved on half-strength (1/2) MS medium containing 0.5 mg L?1 NAA. Rooted plantlets were hardened under control conditions and successfully acclimatized under field conditions. 相似文献
10.
We examined the effect of zeatin on the formation of shoot buds from explants and callus tissues derived from stem segments of Actinidia polygama Miq. (matatabi or silver vine). Stem and petiole segments cultured on combined broad-leaved tree medium and woody plant medium (BW medium) supplemented with zeatin for 40 days formed many shoot buds. Callus tissues derived from the stem segments and subcultured on BW medium supplemented with 6-benzyladenine and 1-naphthaleneacetic acid formed shoot buds when the medium contained 13.7µM zeatin. BW medium containing low concentrations of sucrose strongly induced the formation of shoot buds from the callus. 相似文献
11.
Aseesh Pandey 《Journal of Sustainable Forestry》2013,32(6):590-603
An efficient micropropagation protocol has been developed for tasar oak. Nodal segments from in vitro grown seedling were used for shoot multiplication. Best shoot multiplication response, in terms of number of shoots per explant as well as shoot length, was obtained in woody plant (WP) medium supplemented with 6-benzyladenine and indole-3-acetic acid (8.88 μM BA + 1.43 μM IAA); but the establishment of cultures was difficult due to basal callus formation and necrosis in due course of time. Out of the two used growth adjuvants, casein hydrolysate (CH, 500 mg L?1) promoted shoot multiplication rate significantly in comparison to silver nitrate and also eliminated the basal callus formation problem and necrosis faced during the later stage of shoot proliferation. In vitro rooting on WP medium supplemented with 100 μM indole-3-butyric acid (IBA) when applied for 48 hr gave the best results in comparison to prolonged exposure. Well-acclimatized plantlets were transferred to the field with 80% survival rate. This protocol could be useful not only to propagate and conserve this oak but can also uplift the socioeconomic status of the Himalayan people as its leaves are used to feed the tasar silk worm during rearing period. This method will also be helpful for propagation of high value trees for a reforestation program. 相似文献
12.
《林业研究》2017,(1)
We developed a shoot multiplication protocol for Syringa reticulata Blume var. mandshurica Hara from in vitro cultured seedlings that derived from in vitro germinated seeds. The shoots could be induced on Murashige and Skoog(MS) medium with proper plant growth regulator combinations of 6-benzylaminopurine(BA) and indole-3-butyric acid(IBA). The better medium for shoot multiplication and growth was MS + 5 mg L~(-1)BA +0.5 mg L~(-1)IBA + 20 g L~(-1)sucrose +7 g L~(-1)agar, and the corresponding shoot induction rate was 75 %. The plantlets grew well after rooting on 1/2MS medium(macro-elements of MS medium are at half-strength) supplemented with 1 mg L~(-1)IBA, and the survival percentage was 80 % at 16 weeks after transplanting. 相似文献
13.
白刺花胚性愈伤组织诱导及体细胞胚发生 总被引：1，自引：0，他引：1
【目的】探讨不同植物生长调节剂对白刺花胚性愈伤组织诱导的作用,以及培养基中氮源和无机盐浓度对白刺花体细胞胚发生和植株再生的影响,以期建立白刺花体细胞胚发生、发育及调控技术体系,为白刺花种苗快速繁殖体系建立及遗传转化研究提供参考。【方法】以白刺花叶片为外植体,研究生长调节剂2,4-D(1.0、2.0、3.0、4.0 mg ·L -1 )、NAA(0、0.5、0.8、1.0 mg ·L -1 )、6-BA(0.2、0.5、1.0、2.0 mg ·L -1 )和TDZ(0、0.2、0.5、1.0 mg ·L -1 )组合对胚性愈伤组织诱导,及NAA(0、0.2、0.5 mg ·L -1 )、6-BA(0、0.5、1.0 mg ·L -1 )和TDZ(0、0.2、0.5 mg ·L -1 )组合对体细胞胚发生的调控作用,筛选最优生长调节剂组合;并研究培养基中KNO 3和NH 4NO 3比例对体细胞胚发生的作用,及MS培养基中无机盐浓度(1/5MS 、1/4MS、1/3MS、1/2MS)对体细胞胚萌发的影响,筛选最佳的体细胞胚发育及成熟萌发条件。【结果】白刺花叶片外植体胚性愈伤组织诱导适宜培养基为MS + 2,4-D 3.0 mg ·L -1 + NAA 0.5 mg ·L -1 + 6-BA 0.2 mg ·L -1 + TDZ 1.0 mg ·L -1 +蔗糖40 g ·L -1 +琼脂7.0 g ·L -1 ,诱导率为42.0%。采用MS基本培养基时,最佳的体细胞胚发生培养基为MS + NAA 0.5 mg ·L -1 + 6-BA 1.0 mg ·L -1 + TDZ 0.5 mg ·L -1 +蔗糖40 g ·L -1 +谷氨酰胺100 mg ·L -1 +琼脂7.0 g ·L -1 ,体细胞胚发生率为78.46%,总胚数为对照的3.6倍;MS培养基中,KNO 3浓度提高1倍、NH 4NO 3降至1/2时,体细胞胚发生率可提高至91.33%,总胚数为采用MS基本培养基时的1.4倍;1/3MS培养基有利于体细胞胚的萌发,萌发率为82.75%,幼苗长势良好,单株平均鲜质量为76 mg,幼苗驯化移栽1个月后成活率达90%以上。【结论】白刺花叶片接种于添加2,4-D、NAA、6-BA和TDZ不同组合的诱导培养基上,可脱分化形成愈伤组织或胚性愈伤组织,2,4-D浓度对愈伤组织形态和质地有较大影响。TDZ有利于体细胞胚的形成,适宜浓度的生长素与细胞分裂素组合及硝态氮和铵态氮的比例对体细胞胚的形成和发育具有调控作用,降低MS无机盐浓度可提高体细胞胚萌发率,本试验体系的再生植株移栽成活率达90%以上。 相似文献
14.
Cottonhead windhairdaisy (Saussurea laniceps Hand.-Mazz.) is one of the most famous and important medicinal herbs in China. Illegal collection from wild populations is increasingly threatening the present environment of S. laniceps. Establishment of an efficient method for micropropagation is the best way to change its endangered situation. When mature seeds of S. laniceps were cultured on hormone-free MS medium, plantlets were formed from germinated seeds in 7–10 d. Then 0.5 cm × 0.5 cm leaf explants were transplanted to MS medium supplemented with 1-naphthalene-acetic acid (NAA)/2,4-D and benzyladenine (BA)/KT and callus was achieved 10 d after transfer. Shoot bud regeneration occurred from callus cultured on MS medium supplemented with different growth regulators 20 d after culturing. The regeneration percentages varied with the different components of plant growth regulators. The percent regeneration from callus pretreated at low temperature of 5°C increased significantly compared with those incubated at 23/20°C directly. Optimal regeneration was observed with explants on media supplemented with 1.5 mg•L–1 BA plus 0.2 mg•L-1 NAA. In the presence of 0.2 mg•L–1 NAA in half-strength MS, 78% of the shoots formed roots. Plantlets from explants showed 63% survival after acclimatization. 相似文献
15.
Qingbin Jiang Yong Zhang Chonglu Zhong Bingshan Zeng Didier Bogusz Claudine Franche 《New Forests》2012,43(2):143-154
An in vitro plant regeneration protocol via indirect organogenesis from morphogenetic callus was established for Casuarina cunninghamiana Miq. Effects of plant growth regulator NAA (naphthaleneacetic acid) and BAP (6-benzylaminopurine), sucrose and AgNO3 on callus induction, adventitious bud differentiation and shoot development were examined. Explants used were epicotyl fragments
from 45-day-old seedlings. The largest callus (4.29 mm in diameter) was obtained after 1 month on a basic culture medium consisting
of Murashige and Skoog ? macro- and full strength micro- elements, Nitsch and Nitsch vitamins, supplemented with 0.54 μM NAA,
3.30 μM BAP, and 30 g L−1 sucrose. The calli were subcultured in the same medium above for 2 months. They were then cultured for another 2 months for
adventitious bud differentiation and shoot development. The highest mean adventitious bud differentiation, number of shoots
formed per callus and number of shoots ≥2 cm long per callus (47.50%, 27.38 and 4.75, respectively) were achieved on the above
medium modified with NAA at 0.27 μM and supplemented with AgNO3 1 mg L−1. Shoots were successfully rooted without plant growth regulator and the rooted plantlets survived and grew normally. This
protocol for in vitro plant regeneration provides a tool not only for vegetative propagation but also for plant genetic transformation
and gene function studies of C. cunninghamiana. 相似文献
16.
In vitro regeneration of complete plants from nodal single-bud segments of 2-year-old Australian Cedar (Toona ciliata) trees were obtained under defined nutritional and environmental conditions. Explants were dissected from plants obtained by germination of seeds and growth in pots in a greenhouse. The best medium for shoot regeneration was that of Murashige and Skoog at 1/4 strength with 3% sucrose (1/4 MS), supplemented with 0.1 mg/l IBA and 0.5 mg/l BAP. Rooting of regenerated shoots was observed in MS medium with 0.1 mg/l IBA. Using mature tree material was more difficult. Forced flushing was used to induce shoot development on branches of a 10-year-old tree. Nodal segments of these epicormic shoots formed shoots in vitro on 1/4 MS + 0.01 mg/l IBA + 5 mg/l BAP, but rooting was never observed. 相似文献
17.
柳叶蜡梅叶片愈伤组织诱导实验研究 总被引：1，自引：0，他引：1
以不同方式处理柳叶蜡梅种子，观察其在MS培养基上萌发和生长的状况。结果显示：98％浓硫酸处理的种子发芽率达32％，其它处理萌发率低。以种子无菌苗叶为外植体，接种于2，4-D、NAA、6-BA组合不同浓度生长调节剂的MS固体培养基上，进行愈伤组织诱导实验；外植体愈伤组织诱导培养基MS＋NAA0．3mg·L^-1＋6-BA1．0mg·L^-1的诱导率为80％，生长稳定可分化出苗。 相似文献
18.
以增殖率较低的21号黑木相思无性系继代苗为材料，希望通过培养基调节，提高生产效率，为推动规模化发展打下基础。通过单因子试验，以苗高、新芽数、愈伤质量和苗木生物量为衡量指标，筛选出最佳的大量元素、6-BA、IBA及糖用量。结果表明：培养基中大量元素含量对愈伤质量和生物量影响显著，以MS大量最好；6-BA含量对苗高、新芽数、愈伤质量都产生显著影响，以不加6-BA苗高最高，6-BA 0.5 mg/L 获得最多5.4个新芽/苗，6-BA 1.0 mg/L愈伤质量最大，IBA浓度只对新芽数产生显著影响，以IBA 0.5 mg/L新芽数最多，糖对所有指标产生显著影响，当糖20 g/L时，苗木生长最高，糖20～30 mg/L时，新芽数、愈伤质量和生物量最大。 相似文献
19.
栓皮栎体胚的增殖、成熟和萌发 总被引：1，自引：0，他引：1
以栓皮栎实生苗叶片为外植体诱导体细胞胚胎发生,调查碳水化合物、渗透剂和植物生长调节剂对体胚增殖、成熟和萌发的影响,建立体胚增殖、成熟和萌发的培养基方案.叶片外植体在附加1.0 mg·L-1NAA和0.5mg·L-1BA的起始培养基上诱导形成前胚性团块.这些胚性团块在增殖培养基上培养6周,在附加1 mg·L-1BA、0.25 mg·L-1NAA和3%蔗糖的MS培养基上增殖效果最好.将单个体胚转接到成熟培养基上进行培养,蔗糖浓度对栓皮栎体胚成熟以及后续的萌发有显著影响.成熟培养基中附加5%的蔗糖,体胚成熟率和萌发率分别达到63.5%和33.8%.虽然在成熟培养基中附加ABA有利于体胚成熟,但对体胚的进一步萌发没有促进作用.为了提高萌发率,成熟体胚在附加植物生长调节剂的萌发培养基上进行培养,以及进行预冷处理.成熟体胚4℃冷处理没有促进胚根和上胚轴的发育萌发.在附加0.5 mg·L-1BA和0.25 mg·L-1 IBA的1/2 MS萌发培养基上,体胚萌发率达到65.9%,再生植株转化率达到9.4%. 相似文献