爬行卫矛下胚轴高频离体再生体系的建立(英文)

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.     

不同植物生长调节剂对白木香扦插生根的影响

The rooting ability of the cuttings of semi-lignified branches of Aquilaria sinensis by different treatments were studied in this paper.The treatments included the solutions of IAA,IBA,NAA and ABT-1 with the concentrations of 500,1 000 mg·L-1 and 1 500 mg·L-1.The results showed that there were significant differences in the rooting ratio among different treatments.The rooting ratio of cuttings with IAA 1 500 mg·L-1 treatment was the highest at 43.08%,followed by the cuttings with IBA 1 500 mg·L-1.The advent...     

Regeneration and gene transformation systems of Robima pseudoacacia ''''Idaho'''' mediated by Agrobacterium tumefaciens

Robinia pseudoacacia 'Idaho' is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration. For this paper, we studied the effects of auxin and cytokine on Idaho locust in vitro regeneration and the establishment of gene transformation systems for plants mediated by Agrobacterium tumefaciens. Results showed that the ratios of cytokinin and auxin were the major factors affecting adventitious bud differentiation on a MS medium; the concentration of 0.5 mg·L-1 6-BA benefitted callus proliferation and 0.25 mg·L-1 IBA promoted shoot rooting; however, a higher IBA concentration will inhibit rooting. The most effective antitoxin for screening transgenic Idaho locust shoots was G418 and the most sensitive concentration of it was 8 mg·L-1.     

Effects of antibiotics on plantlet regeneration via organogenesis in Populus euphratica

Four antibiotics, kanamycin, geneticin, carbenicillin and cefotaxime,were evaluated for their effect on the regeneration of adventitious buds, shoot differentiation, rooting from regenerated shoots of Populus euphratica as well as on their control of Agrobacterium -mediated transformations. Results showed that the optimum concentration ranges of kanamycin and geneticin were 15-20 and 10-15 mg·L-1 at the stage of transgenic plantlet selection. The inhibitory effects of cefotaxime and carbenicillin varied among different genotypes of Agrobacterium. The inhibition of cefotaxime on Agrobacterium C58 was stronger than that of carbenicillin. The optimum concentration of cefotaxime was 100 mg·L-1. Cefotaxime and carbenicillin had similar effects on Agrobacterium LBA4404 and their optimum concentrations were both 150 mg·L-1.     

Effect of auxins on axillary and de novo shoot regeneration from in vitro shoot cultures derived from forced epicormic buds of teak (Tectona grandis L.)

In this presentation,we report on de novo and axillary shoot regeneration and rooting of shoots maintained over a long term,from cultures of Tectona grandis L.Shoot-tips of teak shoots forced from epicormic buds were used as the starting material for axenic shoot-culture establishment.Long term maintenance of such axenic shoot cultures was carried out by regular sub-culturing on MS media supplemented with N 6-benzyleadenine (BA,8.8 μmol L-1) and indole-3-butyric acid (IBA,2 μmol L-1) for 24 months.Vigorously growing shoot tips (2 3 cm long) were inoculated on the MS basal medium supplemented with different concentrations (0,1,2,4,6,8 or 10 μmol L-1) of either IBA or α-naphthaleneacetic acid (NAA) for rooting.Axillary and de novo shoots were developed from axillary and cut basal ends of shoots,respectively.Shoots growing on auxins were further sub-cultured (every 15 days) and maintained for 45 days.The greatest number of de novo (5.06) as well as axillary shoots (2.85) was observed on the MS medium supplemented with 10 μmol L-1 NAA or 8 μmol L-1 IBA,respectively,after 45 days.The combinations of both IBA (μmol L-1) + NAA (μmol L-1) were tested at different concentrations (4 + 4,6 + 6,8 + 8) supplemented to a half strength MS basal medium with 0.1% activated charcoal for rooting of decapitated and non-decapitated de novo and axillary shoots.Rooting from non-decapitated de novo shoots was highest (93.33%) with a mean number of roots of 4.61 on this medium,supplemented with 6 μmol L-1 IBA + 6 μmol L-1 NAA,after 36 days of initial culture.Individual auxin,however,was not effective for root induction.Rooted shoots were acclimatized in a green house and after four weeks plantlets were transferred to the field.     

Establishment of in vitro culture of Populus euphratica Olivier

The purpose of our study was to establish a regeneration system for micropropagation of Populus euphratica Olivier. On the basis of an analysis of plant leaf mineral nutrients, a special medium was proposed, called MP2. In optimizing media for in vitro plant cultures including MS, B5 and MP2 media we employed hormones, auxin IAA, cytokine benzyladenine (BAP) and gibberellic acid (GA) in our factorial experiments on media. Adventitious shoots were derived from cuttings of adult plants taken from Xingjiang, west China, on selected media with MP2 0.5 mg·L-1 BA 0.1 mg·L-1 NAA. The shoots were elongated on a medium with 0.25 mg·L-1 BAP, 0.1 mg·L-1 NAA and 2 mg·L-1 GA and were then rooted on a medium with 0.2-0.5 mg·L-1 IBA. All the media were incorporated with 30 g·L-1 sucrose and an adjusted pH at 6.3.     

Low temperature, IBA concentrations and optimal time for adventitious rooting of Eucalyptus benthamii mini-cuttings

Eucalyptus benthamii is a forest species of economic interest that has difficulty with seed production and also is considered to have difficulty with adventitious rooting using propagation techniques,such as cutting or mini-cutting.We aimed to assess the adventitious rooting percentage under different storage times in low temperatures and at various IBA(indole-3-butyric acid) concentrations to determine the optimal time of permanence for rooting Eucalyptus benthamii minicuttings in a greenhouse.Shoots collected from mini-stumps cultivated in a semi-hydroponic system were used to obtain the mini-cuttings.For the first experiment,the mini-cuttings were stored at 4℃ for 0(immediate planting),24,48,72,96 and 120 h.The second experiment evaluated the rooting dynamic to determine the optimal time of permanence for minicuttings in a greenhouse.The basal region of the mini-cutting was treated with various IBA solutions:0(free of IBA),1,000,2,000,3,000 and 4,000 mg·L-1.Every seven days(0(immediate planting),7,14,21 and 28 days),destructive sampling of the mini-cuttings was performed to evaluate the histology of the adventitious rooting.Eucalyptus benthamii minicuttings should be rooted immediately after the collection of the shoots.The 2,000 mg·L-1 IBA concentration induced a greater speed and percentage of adventitious rooting,and an interval of 35 to 42 days was indicated for permanence of the mini-cuttings in the greenhouse.Exposure to low temperature induced adventitious root formation with diffuse vascular connections.     

Establishment of Efficient Regeneration System of Populus×euramericana cl.‘Bofeng’

Compared with the species of Leuce section,the difficulty in plant regeneration during tissue culture for the species of Aigeiros section is a key limiting factor for their application in genetic engineering of forest trees.In this study,the plant growth regulators combination,copper concentration,light intensity,and the selection pressures of kanamycin were investigated using leaves of Populus×euramericana cl.’Bofeng’ as explants,and a stable and high efficient regeneration system was established.Using this system,both the shoot regeneration rate and rooting rate were up to 100%,and the average number of differentiated shoots in each leaf explant was up to 20,and the survival rate of seedlings reached 98%.The optimal selection pressure of kanamycin in shoot inducement for leaf-explant was 40 mg·L^-1,and the optimal selection pressure of kanamycin in root inducement of adventitious bud was 20 mg·L^-1.The authors also found that differentiation rate of adventitious buds of P.×euramericana cl.’Bofeng’ could be significantly promoted by increased copper concentration,and this is the first report about the key role of copper（Cu） in explant regeneration of Poplars.This regeneration system with high frequency which was established by the authors was as good as that of species of Leuce section,and it provides a good technological platform for genetic engineering of forest tree which uses the species of Aigeiros section as model materials.     

Vegetative propagation of Dalbergia sissoo: effect of growth regulators, length, position of shoot and type of cuttings on rooting potential in stem cuttings

The effect of auxins (IAA or IBA at 100,200,500 mg L 1) on rooting and sprouting parameters differed significantly (p < 0.05).Rooting and sprouting percentages were higher in some treatments while other parameters were significantly higher in other treatments.The results of the present study revealed that rooting was significantly (p < 0.05) affected by the length of cuttings,as well as the position of shoots and type of cuttings.On average,86.0 per cent rooting was observed in the 30-cm long cuttings compared to the other two lengths of cuttings,i.e.,20 and 25 cm,irrespective of any auxin treatment.In a control set,without any auxin treatment,a maximum of 60.0% rooting was recorded in the 30-cm long cuttings and a minimum of 40.0% of the cuttings rooted in the 20-cm long cuttings.Similarly,cuttings collected from the lower and upper portions of shoots resulted in 72.5 and 52.5 per cent rooting respectively,irrespective of auxin treatments.Maximum rooting was observed in the lower portion (40.0%) of cutting compared to the upper portion (30.0%) in the control set without auxin treatment.Leafless branches produced maximum (72.0%) rooting compared to the leafy cuttings (35.0%),irrespective of the auxin treatment.The leafy cuttings produces a minimum of 15.0% sprouting and rooting in the control set.     

Preservation of Juniperus thurifera L.:a rare endangered species in Algeria through in vitro regeneration

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.     

长白山区珍稀濒危植物野生玫瑰植株再生体系的建立

玫瑰(Rosa rugosa Thunb.)又称玫瑰花,是蔷薇科 (Rosaceae) 蔷薇属(Rosa L.)落叶灌木.该种为国家三级重点保护植物.<中国珍稀濒危保护植物名录(第一册)>中定为濒危种,<中国物种红色名录>定为易危种[1],<吉林省野生动植物保护管理暂行条例>中定其为省级一类重点保护植物.是待开发的野生珍稀濒危药用植物,花、果、根入药,主治吐血、痛经、咯血、乳痈、月经不调等症.该种玫瑰具有喜光,耐寒,耐贫瘠,抗风等特性,可防止高山苔原带的水土流失,对维护生态平衡,美化天池环境以及引种驯化、开发利用于作护坡保土、园林绿化和培育玫瑰新品种;对研究该属植物的系统发育与地理分布也有重要科学价值.玫瑰在长白山区数量非常稀少,仅在珲春市敬信乡有零星分布.近年来,由于旅游设施及道路的修建等人为因素的干扰,其生境遭到严重破坏.又因玫瑰的天然更新能力非常弱,其种子成熟度小,种子繁殖可操作性极差,插条繁殖生根率和移栽成活率也极低,使其开发及利用受到极大限制.因此,在保护好现有野生资源的同时,本研究以玫瑰新生嫩茎为外植体,利用植物组织培养方法,采用均匀设计法,以期筛选出玫瑰嫩茎离体培养各阶段的培养基.目前,与其同属的其他种植物的离体培养已有报道,但关于该种玫瑰植株再生体系的研究迄今未见.     

白花泡桐优树试管嫁接幼化及组培快繁技术研究

以白花泡桐优树‘白优2号’为试验材料,通过组织培养和试管嫁接方法,对白花泡桐优树材料的幼化技术进行了研究.结果表明:外植体初代培养萌发的嫩芽为最适合的接穗;‘建始桐3号’为试管嫁接较合适的砧木;采用劈接进行;MS+ NAA0.3 mg·L-1+蔗糖30 g·L-1为试管嫁接培养基;继代增殖和生根培养基分别为1/2MS ...     

马占相思优树组培快繁技术研究*

相思(Acacia Mill.)类树种(俗称相思树)属含羞草科(Mimosaceae)金合欢属(Acacia Mill.),在我国广东、海南、广西、福建、云南和江西等地均有栽培。相思树植株高大挺拔,根系发达具根瘤,具有耐干旱贫瘠及抗风的特性,是水土保持及改良土壤的优良树种;同时,相思树还是优良的用材树种,其生     

中国特有濒危植物崖柏扦插繁殖研究

在重庆市城口县咸宜乡苗圃和中国林科院温室开展崖柏扦插繁殖试验,研究采穗母树年龄、不同激素种类及其组合、激素浓度、扦插基质、物理促根等不同处理方式下崖柏扦插生根效应.结果表明:(1)在咸宜乡苗圃,采用自动喷雾设施,以蛭石-泥炭土-珍珠岩(1∶1∶1)为扦插基质,用3种激素和清水处理崖柏幼树1年生枝条,扦插后160d的生根率为74.76％～96.70％,不同处理组合生根效应具有显著差异,2000 mg· L-1 IBA＞清水对照(CK)＞1 000mg·L-1 IBA＞1 000 mg·L-1IBA+ 500 mg· L-1NAA;(2)在林科院温室用2种激素和清水处理崖柏幼树1年生枝条,扦插于蛭石-泥炭土-珍珠岩(1∶1∶1)的基质,搭塑料拱棚,扦插后120 d的生根率远低于咸宜乡苗圃,为25.93％～45.37％,激素处理后的枝条与对照差异显著,1 000 mg·L-1 IBA+ 50 mg·L-1 NAA=2 000 mg·L-1 IBA＞清水对照(CK);(3)在咸宜乡苗圃用不同激素和物理促生根方法组成的27种组合处理的成年崖柏枝条,最高生根率仅为20.83％.生根率超过10％的处理组合是:清水处理,泥炭土∶蛭石∶珍珠岩(1∶1∶1);1 000 mg·L-1IBA+500 mg·L-NAA,泥炭土∶珍珠岩(2∶1);1 000 mg·L-1IBA,泥炭土∶蛭石∶珍珠岩(1∶1∶1),流水冲洗.     

长蕊杜鹃扦插内源激素变化及解剖结构观察

以野生长蕊杜鹃半木质化茎段为试材,采用植物生长激素KIBA,KNAA和Hormodin对插穗进行处理,测定不同生根期内源激素IAA、GA3、ZR、ABA含量,对扦插不同时期插穗茎段进行电镜扫描。结果表明:先快蘸液态KNAA(3 000mg·L-1),再迅速蘸取粉状Hormodin#1处理对插条生根具有显著影响,插穗生根率达到了81.25%;较高浓度的内源IAA含量有利于根原基分化形成,GA3含量上升与插穗愈伤组织的诱导、不定根的形成呈正相关性,低水平的ABA更有利于长蕊杜鹃扦插生根;插穗在根原基分化和愈伤组织形成时期消耗內源ZR,在不定根长出后自身合成ZR;通过对淀粉粒研究表明扦插期补充糖类物质是必要的,簇状晶体产生表明钙调控等生理机制形成。     

麻疯树叶盘法高效再生的研究

以温室中1年生麻疯树顶端幼嫩叶片为外植体,研究了在MS基本培养基中添加不同浓度的6-苄基腺嘌呤(6-BA)和吲哚丁酸(IBA)对不定芽再生的影响,并采用60天暗培养的方法筛选出愈伤组织诱导和不定芽诱导的最佳激素组合为MS+5 mg.L-16-BA+0.5 mg.L-1IBA,该组合的不定芽诱导率高达75.8%。将该组合诱导出的愈伤组织接种至MS+1.5 mg.L-16-BA+0.05 mg.L-1IBA的固体培养基中,研究了赤霉素对不定芽再生的影响,最佳赤霉素浓度为0.05 mg.L-1,麻疯树叶盘再生率达到90.9%,平均不定芽个数达到4.6个。     

卷荚相思组培快繁技术研究

[目的]建立16年生卷荚相思优树组培快繁技术体系。[方法]以16年生卷荚相思优树的当年新生枝条带腋芽茎段为材料,对卷荚相思外植体进行消毒、初代培养、增殖培养、生根培养和移植等。[结果]表明:通过对16年生卷荚相思成年优树采条进行扦插,以扦插苗建立采穗圃,选取采穗圃中当年生健康无病虫害枝条的中段为外植体,最佳消毒方式为75%的酒精处理0.5 min和0.1%的升汞处理18 min,其存活率达69.33%,芽诱导率达86.67%;最佳初代培养基为改良MS+蔗糖40 g·L-1,出芽率为91.33%。最佳增殖培养基为MS+6-BA 0.5 mg·L-1+NAA 0.1 mg·L-1+蔗糖30 g·L-1,35 d增殖倍数可达3.50倍;最佳生根培养基为1/2 MS+IBA 0.25 mg·L-1+NAA 0.5 mg·L-1+蔗糖40 g·L-1,15 d生根率为96.11%;将生根苗移植至以沙为基质的营养杯中,存活率为71.11%。[结论]研究解决了16年生卷荚相思成年优树外植体污染率高和芽诱导率低等问题,建立的组培快繁技术体系对今后加快卷荚相思良种选育及优质苗木大量扩繁具有重要作用。     

穗条生根剂育苗基质和季节对西南桦扦插生根的影响

西南桦(Betula alnoides Buch.-Ham.ex D.Don)是我国西南省(区)造林规模最大的乡土珍贵用材树种.西南桦具有材性优良、容易加工、色泽淡红、不易变形等木材特性,是高级家具、室内装修、木地板的优质材料,经济价值甚高.由于目前育苗用种多采自天然次生林,随着西南桦天然资源的日益贫乏,加剧了造林规模不断扩大与供种量逐渐减少的矛盾;而且,实生苗造林的林木个体间遗传变异较大,林分木材单产较低.随着良种选育工作的深入,西南桦人工林向"有性选育,无性利用"路径发展.     

激素对复叶槭茎段和叶片愈伤组织诱导及再生的影响

以复叶槭的茎段、叶片为外植体,以MS为基本培养基,附加多种激素配比试验,结果表明:叶外植体在各种培养基上均形成愈伤组织,但没有出现芽的分化。茎段外植体在多种培养基上均产生愈伤组织,但仅在MS+0.000 5mg.L-1TDZ+0.01 mg.L-1NAA培养基上出现再生芽,芽的分化率为22.8%,在MS+0.1 mg.L-1NAA上生根率为83.3%,移植后成活率为86.2%。