Micropropagation of <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> × <Emphasis Type="Italic">E. urophylla</Emphasis> AEC 224 clone

Genetic transformation systems require protocols that allow regenerating transgenic plants from transformed tissues. This study aimed to establish a protocol for indirect organogenesis in leaf explants of a Eucalyptus grandis  ×  E. urophylla AEC 224 clone. During callogenesis stage, several concentrations of NAA and then NAA or 2,4-D combined with TDZ were tested in JADS culture medium for 30 days, followed by subculture of the explants in the regeneration medium, containing 5.0 µM BA and 0.5 µM NAA for another 30 days. In these media, the explant oxidation rate was high (95 %). Thus, in order to reduce oxidation, different culture media were compared: WPM, MS, JADS and modified QL, followed by explant transfer onto regeneration medium. The highest percentage of regeneration and the lowest oxidation rate were achieved on WPM medium. Then, NAA and 2,4-D were tested in combination with TDZ and also TDZ and BA combined with NAA in WPM medium. The most efficient culture media in terms of shoot regeneration were WPM supplemented with 0.25 µM TDZ and 0.1 µM NAA during 30 days for callus induction and then with 5.0 µM BA and 0.5 µM NAA for another 30 days. This protocol yielded a regeneration rate of 43 %, with a low oxidation of tissues. A rooting experiment was conducted using half strength MS medium and comparing three concentrations of IBA (2.46, 4.90 and 7.35 µM). The highest rooting percentage (35 %) was obtained on medium containing 2.46 µM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %.     

显脉金花茶无菌体系建立及增殖培养研究

以显脉金花茶幼嫩带芽茎段作为外植体,开展外植体灭菌、启动培养及增殖培育试验,结果表明:3月采集的外植体最易灭菌,其最佳灭菌方案为先用75%酒精浸泡20 s后用0.1%升汞浸泡9 min;最佳启动培养基配方为WPM+6-BA 7 mg.L-1+NAA 0.25 mg.L-1;最佳增殖培养基配方为:改良WPM(NO3-∶NH4+=2∶1)+6-BA 5 mg.L-1+NAA 0.05 mg.L-1。     

火焰卫矛组织培养及植株再生研究

以火焰卫矛成熟胚培养长出的子叶为外植体进行组织培养及植株再生研究,建立了火焰卫矛再生体系,成熟胚培养基为WPM+TDZ 0.5 mg.L-1;子叶愈伤组织诱导和不定芽分化培养基为WPM+6-BA 6 mg.L-1+IBA 0.2 mg.L-1;生根培养基为1/2MS+NAA 0.2 mg.L-1+活性碳0.5 g.L-1。     

铁皮石斛原球茎增殖条件的研究

研究了不同基本培养基、不同浓度的脱落酸和培养方式对铁皮石斛原球茎增殖的影响。结果表明在改良MS中加入0.5 mg.L-1脱落酸,进行固体培养或液体振荡培养,原球茎增殖的效果都比较好。     

乌桕茎段诱导组培快繁技术研究

通过对乌桕茎段诱导组培快繁技术研究结果表明:萌孽条中部未木质化部分的茎段是理想的外植体材料,采用含6%有效氯的次氯酸钠溶液与0.1%HgCl2溶液交替消毒,外植体灭菌效果好,存活率高;适合愈伤组织诱导的培养基为MS+6-BA 0.5 mg.L-1+2,4-D 1.0 mg.L-1;适合不定芽分化与增殖的培养基为MS+6-BA 2.0 mg.L-1+NAA 0.1 mg.L-1,分化系数达4.68;不定芽在1/2MS+IBA 0.5 mg.L-1+NAA 0.2 mg.L-1培养基上生根效果最好;生根苗移栽到珍珠岩、蛭石混合基质中,成活率可达90%以上。     

牡丹愈伤组织的诱导及愈伤褐化抑制的研究

以凤丹为试验材料,比较不同外植体,不同基本培养基,不同激素组合对愈伤诱导率的影响;结果表明,以子叶和胚轴为外植体,在培养基1/2改良WPM+2,4-D1.0 mg·L-1+6-BA1.0 mg·L-1+水解酪蛋白500mg·L-1+L-谷氨酰胺150 mg·L-1+蔗糖30g·L-1+琼脂7g·L-1的培养基中诱导率最高,可达100%。同时研究了愈伤褐化抑制,结果表明,以1/2改良WPM为最佳基本培养基;PVP和植物凝胶的效果最好,褐化指数分别为31.11%与30.00%;黑暗、低温、较短的继代周期都能不同程度的减缓愈伤褐化。     

速生优良杉木组培快繁技术试验

应用正交设计法研究外植体的最佳消毒方式,同时考察了培养基、6-BA、KT、NAA等4个因子对杉木组培苗芽增殖的影响和IBA、NAA、ABT1#、蔗糖等4个因子对组培苗生根培养的影响。结果表明:以茎尖为外植体材料,消毒方式采用70%乙醇处理40 s后无菌水冲洗一遍,再用0.1%升汞溶液处理7 min后用无菌水冲洗5次以上;最佳的增殖培养基是1/2MS+6-BA 0.8 mg·L-1,最适于生根的培养基是MS培养基中+蔗糖30 g.L-1+IBA 1.2 mg·L-1+NAA 0.2 mg·L-1+ABT1#1.5 mg·L-1。     

Modulation of in vitro morphogenesis in nodal segments of Salix tetrasperma Roxb. through the use of TDZ, different media types and culture regimes

A comparative performance of two different media formulations (woody plant medium (WPM) and Murashige and Skoog??s (MS) medium) for their ability to inflict in vitro shoot development in nodal segments of Salix tetrasperma Roxb. has been carried out. Thidiazuron (TDZ) in various concentrations was used as a supplement to the basal media. Media types, TDZ concentrations, exposure duration and culture regimes played an important role in affecting multiple shoot production. WPM supplemented with 2.5???M TDZ for 4?weeks exposure was found to be the best for maximum (4.53?±?0.27) shoots production in vitro. Transfer to a secondary medium consisting of 6-benzyladenine (1.0???M) and ??-naphthalene acetic acid (0.5???M) enhanced the multiplication rate and by the end of 12?weeks, 20.33?±?0.33 shoots with shoot length, 4.70?±?0.26?cm were produced on WPM. Rooting of the regenerated shoots was achieved on half strength basal media (either WPM or MS) containing 0.5???M indole-3-butyric acid. In all the experiments, different growth parameters were scored and WPM was found to be superior to MS medium. The regenerated plantlets were successfully acclimatized in the field with about 81?% survival.     

赤桉组培快繁技术研究

以赤桉优株的带芽茎段为外植体进行组织培养技术研究,成功建立了赤桉组培快繁体系。研究结果表明：改良MS是赤桉腋芽增殖启动培养的适宜培养基,启动率为77.3%。改良MS＋6-BA1.5mg·L-1＋NAA0.1mg·L-1是赤桉继代增殖培养的适宜培养基,20d后增殖系数可达4.17,平均芽长2.9cm。改良1/2MS＋IBA1.0mg·L-1＋NAA0.5mg·L-1是赤桉生根培养的最适培养基,生根率达99.5%。3000～6000lux是赤桉增殖的最适光照强度,1500～3000lux是其生根的最适光照强度。     

史密斯桉组织培养技术研究

本文探讨了史密斯桉组织培养技术。试验表明,经HgCl2消毒后用1%的氯化钙浸洗处理,能有效控制外植体褐化的发生,褐化死亡率仅为12.5%;始芽萌动以及初期继代培养阶段,采用F＋6-BA 0.5 mg.L-1＋IBA 0.3mg.L-1培养基,有利于外植体的不定芽诱导以及初期的继代扩大培养,增殖系数达3.7;在增殖与伸长生长阶段,采用改良H＋6-BA 0.125～0.25 mg.L-1＋IBA 0.25 mg.L-1＋VC 20 mg.L-1＋VB210 mg.L-1培养基,在保证较高增值系数的情况下,有利于芽的伸长生长;以1/3 MS为基本培养基,添加0.25 mg.L-1IBA对史密斯桉的根系诱导较为有效,生根率达37.8%。     

柠檬桉的组培快繁技术研究

以柠檬桉优株带芽茎段为外植体进行组培快繁技术体系研究。结果表明：晴天＋流水冲洗1h＋0．1％HgCl2（3＋4）min为柠檬桉的最佳外植体采集处理方案。培养基添加VC20．0mg.L-1或PVP2．0g.L-1是褐变控制的有效措施,不同防褐化剂的效果为：PVP=VC〉AC〉DTT〉CA。改良MS＋6-BA1．0mg·L-1＋KT0．2mg·L-1＋NAA0．05mg.L-1是柠檬桉扩繁增殖的最适培养基;改良MS＋6-BA0．5mg.L-1＋NAA0．1mg.L-1是柠檬桉壮苗增殖的最适培养基。自然光培养,光照强度3000～3500Lux是柠檬桉增殖培养的最适光照条件。     

龙芽葱木的组织培养及快速繁殖的研究

以龙芽葱木的芽为外植体进行组织培养试验,经试验对比筛选出龙芽葱木的外植体适宜诱导培养基为MS+6-BA1.0 mg.L-1+NAA 0.1 mg.L-1+Vc 30 mg.L-1,继代茎芽分化培养基为MS+6-BA 1.0 mg.L-1+ZT 0.1 mg.L-1+Vc50 mg.L-1,生根培养基为1/2MS+NAA 0.5 mg.L-1,以蛭石+森林腐殖土(1∶1)为培养基质移栽效果好。     

海南龙血树组织培养快速繁殖技术研究

以海南龙血树优株顶芽和茎段为外植体,通过不同细胞分裂素、生长素以及培养基不同浓度的组合对诱导芽的分化、增殖、伸长及生根的对比实验及试管苗移栽管理,筛选出MS 蔗糖30 g/L 6-BA 3．0 mg/L NAA 0．01 mg/L培养基诱导芽的形成,MS 蔗糖30 g/L 6-BA 2．0 mg/L NAA 0．01 mg/L培养基用于芽的增殖,MS 蔗糖30 g/L 6-BA 0．5 mg/L培养基用于壮芽伸长;1/2MS 蔗糖15 g/L NAA 0．2 mg/L用于诱导芽生根;选用泥炭土、椰糠和珍珠岩混合基质,于晚春和秋季移栽试管苗,成活率达90％左右,达到工厂化苗木生产的要求。     

大岩桐组培育苗试验

以大岩桐幼嫩叶片、老叶、带腋芽的茎段为外植体,进行试管诱导,并进行继代、生根、移栽试验.结果表明:大岩桐诱导外植体以带腋芽的茎段效果最好;适宜的启动培养基、增殖培养基和生根培养基分别为MS+6-BA 1.0mg· L-1+NAA0.2mg·L-1、MS+6-BA0.5 mg·L-1 +NAA0.1 mg·L-1和1/2...     

黄金宝树组培育苗技术研究

以黄金宝树基部半木质化萌芽条为外植体,MS为基本培养基,探讨6-BA、KT、NAA对黄金宝树组培育苗的影响,试验结果表明,黄金宝树组培育苗最佳培养基配方分别为:初代培养基MS+6-BA 0.3 mg.L-1+KT 0.5 mg.L-1+NAA 0.3mg.L-1+蔗糖30 g.L-1+卡拉胶8 g.L-1、增殖培养基MS+6-BA 0.5 mg.L-1+KT 0.3 mg.L-1+NAA 0.3mg.L-1+蔗糖30 g.L-1+卡拉胶8 g.L-1、生根培养基1/2 MS+NAA 0.5 mg.L-1+蔗糖20 g.L-1+卡拉胶8 g.L-1。初代培养诱导率达80%以上,有效芽条增殖率达3倍以上,生根率达95%以上,移栽成活率达90%以上。     

两种优良巴西杂交桉树的组织培养和快速繁殖

本研究以巴西尾巨桉和巨赤桉两种优良杂交桉树的带芽茎段为外植体进行离体培养和快速繁殖研究.结果表明:尾巨桉和巨赤桉再生芽诱导的最佳培养基为改良MS+6-BA 0.5 mg·L-1+NAA 0.2 mg·L-1,继代增殖的最适培养基为改良MS+6-BA 0.5 mg·L-1+NAA 0.15 mg·L-1.芽诱导和继代增殖中6-BA的浓度对两种杂交桉树有显著影响.生根培养中,尾巨桉和巨赤桉生根情况差异显著,尾巨桉茎芽最适生根培养基为1/2MS+NAA0.2 mg·L-1+IBA 1.0 mg·L-1,巨赤桉茎芽最适生根培养基为1/2MS+NAA 0.15 mg·L-1+IBA 1.0 mg·L-1.两种杂交桉树组织培养体系的建立,为桉树良种选育和遗传转化体系的研究提供技术参考.     

五味子组织培养技术初探

以五味子带芽鳞的休眠芽、不带芽鳞的休眠芽和水培的新梢茎段为外植体,接种于不同激素配比的培养基中,进行组织培养快繁技术研究。结果表明,水培的新梢茎段是诱导丛生芽的最佳外植体,在MS 6-BA1.5 mg.L-1 IBA0.2 mg.L-1、pH值5.8的初代培养基中,培养效果最为明显,分化率达到了95%。     

钟花樱组织培养繁殖研究

以钟花樱(Cerasus campanulata)的嫩枝作外植体进行组织培养技术研究。结果表明：适合不定芽诱导的培养基为 MS+6-BA 1.0 mg · L-1+NAA 0.1 mg · L-1+蔗糖30 g · L-1+卡拉胶6.5 g · L-1,诱导率为44.44%,适合增殖培养的培养基为 MS+6-BA 1.0 mg · L-1+NAA 0.05 mg · L-1+GA30.5 mg · L-1+蔗糖30 g · L-1+卡拉胶6.5 g · L-1,增殖倍数为3.70,适合生根培养的培养基为1/2 MS+IBA 1.5 mg · L-1+NAA 0.1 mg · L-1+蔗糖30 g · L-1+卡拉胶6.5 g · L-1+活性碳0.5 g · L-1,生根率为82.23%。     

Embryo culture is an efficient way to conserve a medicinally important endangered forest tree species Strychnos potatorum

The present study reports a protocol for germination of Strychnos potatorum (ver. Tel. Chilla) using zygotic embryo culture as an embryo rescue method. A 100% germination rate was obtained by culturing the embryos on full-strength Murashige and Skoog’s medium (MS) containing 20 g/L sucrose in comparison to McCown and Lloyd’s Woody Plant Medium (WPM). Germination rates decreased when the sucrose concentration was lower or higher than 20 g·L-1 . WPM/MS medium containing glucose at levels 30, 20, 15 g·L-1 showed a smaller percentage of germination and at quarter strength, WPM/MS medium with glucose did not respond. Multiple shoot formation was found at 1.0 2.0 mg/L BAP; 3.0 mg/L Kn; 2.0 mg/L TDZ on MS medium with 20 g·L-1 sucrose. Germination rates improved when the embryos were placed upright (vertically) in the medium. The in vitro germinated seedlings were acclimatized in a walk-in-chamber and maintained in the green house with the survival rate of 65% 75%. These plants were transferred to the field and were found to be phenotypically normal, healthy and similar to donor plants. This protocol will be useful to overcome seed dormancy and for rapid multiplication and conservation of S. potatorum using zygotic embryo culture.     

培养基和蔗糖对非洲菊增殖及其生长特性的影响

为提高非洲菊增殖培养效果,增强种苗质量,减少后代变异,在较低植物生长调节剂浓度条件下,研究了不同培养基含量和蔗糖浓度对非洲菊增殖及生长特性的影响情况,并对不同处理后组培苗的增殖系数和生长指标进行了测定。结果表明,在培养基含量为1/4MS~2MS的条件下,苗高、鲜质量、干质量、叶面面积和叶绿素总体上呈现出随培养基含量的增加而提高的趋势;不添加蔗糖,组培苗会停止生长直至死亡,当蔗糖浓度为1.5%~4.5%时,苗高、鲜质量、干质量、叶面面积和叶绿素总体呈现出随蔗糖浓度的增加而提高的趋势,但其增殖系数均表现为先增后降的趋势。据此认为,非洲菊增殖苗培养的最佳条件分别是培养基含量为MS、蔗糖浓度为3%。