桤叶唐棣组织培养研究

以桤叶唐棣的带芽茎段为外植体,对桤叶唐棣离体培养技术进行了初步研究,结果表明：初代培养基为MS IAA0．1mg／L 6-BA 0．5mg／L KT 0．5mg／L;增殖培养基为MS IAA 0．1mg／L 6-BA 2.0mg／L,增殖芽数最大,达到5．4;GA3对茎芽长无促进作用;在1／4MS无激素培养基上,生根率达87．9％。     

剥鳞和激素处理对大樱桃花芽休眠解除及内源激素变化的影响

采用高效液相色谱法(HPLC)分析了剥鳞与激素处理对大樱桃花芽休眠解除及内源生长素(IAA)、赤霉素(GAD、玉米素(ZT)和脱落酸(ABA)变化的影响。结果表明,花芽中的ABA主要分布于鳞片内,鳞片中的GA3和ZT含量远低于去鳞芽,也低于完整芽。剥鳞能明显增加休眠花芽中内源GA2和ZT的含量,降低ABA的含量,对IAA含量的影响不大。剥鳞降低了ABA／GA3、ABA／ZT的比值,使花芽向促进生长、抑制休眠的方向转化。同时,休眠前、后期剥鳞均能明显提高萌芽率,中期剥鳞效果不明显。剥鳞后施用外源激素随休眠时期不同而有不同的破眠效果,早期剥鳞GA3的效果最好,6-BA次之,IAA最差;中期破眠效果不如早期,GA。和6-BA没有明显差别;后期以6-BA效果最好,其次是GA3和IAA;3次处理中ABA均明显抑制花芽萌发。     

百合鳞片组织培养研究

目的:建立百合鳞片组织培养的繁殖技术。方法:研究不同培养基对百合增殖的影响,不同浓度的激素组合对百合鳞片不定芽的诱导、芽的增殖、壮苗和生根的影响。结果:MS培养基为百合最适增殖培养基;最佳诱芽培养基为MS+1.0 mg/L 6-苄基腺嘌呤(6-BA)+0.5 mg/Lα-萘乙酸(NAA),诱导率为62%,苗长势良好;最适增殖培养基为MS+1.0 mg/L 6-BA+0.1 mg/L NAA,增殖倍数最高达3.90,苗健壮,长势良好,叶色浓绿;最佳壮苗培养基为MS+1.0mg/L 6-BA+0.1 mg/L NAA+0.1 mg/L赤霉酸(GA3);最佳生根培养基为MS+0.1 mg/L吲哚丁酸(IBA),平均生根数达4.17。结论:获得了百合鳞片组织培养的最佳培养条件,为百合的资源保护和利用提供了参考依据。     

山竹组织培养技术

本研究以山竹的合子胚为组织培养的外植体材料,研究不同激素浓度配比对山竹得出芽诱导/增殖诱导及生根诱导的影响,以获得一套山竹组织快繁技术,结果表明:GA3能有效提高山竹合子胚的出芽率,最适的山竹合子胚组织培养的基本培养基为WPM;诱导芽的最适培养基为WPM+3.0 mg/L 6-BA+GA31.0 mg/L,增殖最适培养基为WPM+3.0 mg/L 6-BA+1.0 mg/L GA4+7。生根最适培养基为1/2 WPM+1.0 mg/L IBA+1.0 g/L AC。     

甜樱桃(Prunus avium)矮化砧木ZY-1休眠枝条的芽

(1)从生芽诱导培养基:MS 6-BA1.0 mg·L-1(2)增殖培养基:MS 6-BA 1.5 IAA 1.0 GA0.5;(3)壮苗培养基:MS IAA 1.2.     

降低黄芩再生苗玻璃化率及其生根的研究

为了降低黄芩组培苗的玻璃化率并提高其生根率,本研究以黄芩无菌苗茎段诱导的不定芽为实验材料,分别研究了6-苄基嘌呤(6-BA)、蔗糖、琼脂及多效唑(PP333)等组培条件对黄芩不定芽玻璃化的影响以及吲哚丁酸(IBA)对再生苗生根的影响。研究结果表明:低浓度的6-BA有利于降低不定芽的玻璃化率,当培养基中添加0.2 mg·L~(-1)6-BA时,不定芽的玻璃化率最低且增殖系数比对照增加1倍。随着培养基中蔗糖浓度的升高,不定芽的玻璃化率显著降低,其增殖系数也有所降低。培养基中蔗糖浓度为25 g·L~(-1)时,黄芩不定芽长势最好且玻璃化率为零,增殖系数也最高。培养基中添加7.5 g·L~(-1)的琼脂,不定芽的玻璃化率较低,增殖系数也较高。培养基中添加多效唑(PP333)有利于缓解黄芩再生不定芽玻璃化状态,随着PP333的浓度增加,玻璃化率也逐渐降低。0.2 mg·L~(-1)PP333对黄芩不定芽的壮苗起到了很好的作用,再生苗明显变得粗壮。0.1 mg·L~(-1)的IBA最有利于黄芩再生苗的生根,生根率为100%,平均生根数为7,移栽成活率达95%以上。     

不同激素配比对麝香百合鳞茎芽诱导的影响

采用麝香百合的鳞茎片作为外植体进行组织培养,结果表明,不同激素浓度配比对百合鳞茎芽的诱导效果不同,供试的几个组合中,以MS+6-BA 1.0mg/L+NAA 0.5mg/L效果最好;在继代培养中,以MS+6-BA 2.0mg/L+NAA 0.1mg/L对百合鳞茎的增殖效果最好;在生根培养中,附加活性炭可促进根的生长。     

墨兰的无菌播种和植株再生

墨兰的无菌播种结果发现,在不添加细胞分裂素的培养基上,种子可以发芽,但只有原球茎和根状茎产生;不可能进一步分化成苗,只有在含有不同激素成分的MS或KnudsonC培养基上,才有可能诱导芽的分化,其中以附加6-BA 0.5-1.0mg/L+NAA 0.1mg/L诱导效果最佳,在附加6-BA 2.0mg/L+NAA 0.4mg/L的MS培养基中,能加速芽的增殖,根状茎转入含有相同激素成分的液体增殖培养基中振荡培养,可大大提高芽的分化速率。添加0.5%活性炭对芽的分化有明显增效作用。在附加NAA 0.2mg/L的MS培养基中;幼苗的生根效果最佳。     

杂种甜椒的离体快速繁殖（简报）

甜椒F1子叶柄的不定芽诱导率比子叶尖的高;暗培养5d后转入光照下培养,促进不定芽的形成和缩短培养周期;6-BA5.0mg@L-1与IAA0.5mg@L-1组合的不定芽诱导率较高;培养基中添加水解酪蛋白和AgNO3后,不定芽诱导率降低;增殖培养基中添加15%椰汁明显改善试管苗的玻璃化和脱叶现象.     

不同生长调节剂对狗肝菜愈伤组织诱导和离体快繁的影响

目的 :研究不同生长调节剂对狗肝菜愈伤组织诱导和离体快繁的影响。方法 :狗肝菜不同外植体在附加不同生长调节剂的培养基上诱导愈伤组织 ,比较愈伤组织的诱导率 ;用 3因子 5水平的正交实验 ,比较不同生长调节剂对丛生芽诱导的影响 ;在附加不同生长调节剂的培养基上比较芽增殖倍数 ;附加不同浓度NAA的培养基上比较生根效果。结果 :愈伤组织诱导率相对以叶片最高 ,茎段次之 ,最后为叶柄 ;愈伤组织诱导的最佳培养基为MS 6-BA0 .5 NAA1 .5 ;不同激素对茎段芽诱导的影响次序为 6-BA>KT >NAA ,芽诱导的最佳培养基为MS 6-BA2mg/L KT1mg/L NAA0 .5mg/L ;芽继代增殖的最佳激素组合是MS 6-BA2mg/L NAA2mg/L ,增殖倍数达 3.0 0 ,影响芽继代增殖的因素次序为 6-BA >NAANAA0 .5mg/L的生根效果较好。结论 :附加一定的生长调节剂能提高狗肝菜愈伤组织的诱导率和离体快繁的效率。     

Cryopreservation of in vitro Grown Shoot Tips of Red Bud Taro by Encapsulation-Vitrification

A simple procedure for cryopreservation of in vitro grown shoot tips of red bud taro (Colocasia esculenta L. Schott var. cormosus‘Hongyayu’) by encapsulation vitrification is investigated. Shoot tips were excised from 8 week old stock shoots and encapsulated into alginate gel beads. Encapsulated shoot tips were precultured in liquid MS medium supplemented with 35mg·L-1 6 BA, 05mg·L-1 IBA, 01mg·L-1 GA3 and 03mol·L-1 sucrose for 24h, then they were loaded with a mixture of 2mol·L-1 glycerol plus 04mol·L-1 sucrose for 30min at 25℃. After dehydration with PVS2 at 25℃ for 20min, the encapsulated and dehydrated shoot tips were plunged directly into liquid nitrogen. After rapidly rewarming in a 40℃ water bath for 3min, PVS2 was drained from the cryotubes and replaced third with liquid MS medium supplemented with 35mg·L-1 6 BA, 05mg·L-1 IBA, 01mg·L-1 GA3 and 12mol·L-1 sucrose and each kept for 10min at 25℃and then post cultured on solidified MS medium supplemented with 35mg·L-1 6 BA, 05mg·L-1 IBA and 01mg·L-1 GA3 in the dark for 3 days and then transferred to the light conditions. The average survival rate amounted to about 80%. Plantlets regenerated from cryopreserved shoot tips were morphologically uniform. This encapsulation vitrification procedure promises to become a routine method for the cryopreservation of shoot tips of Chinese genuine red bud taro.     

Efficient clonal propagation method for Decalepis hamiltonii, an endangered shrub, under the influence of phloroglucinol

A highly efficient two stage protocol was developed for induction of multiple shoots from single node in vitro shoot tip explants of Decalepis hamiltonii. It was found that phloroglucinol (PG) had synergistic effect on shoot multiplication when added with N6-benzyladenine and gibberellic acid. This protocol uses PG for both multiple shoot induction from nodal explants, elongation of primary shoots and initiation of adventitious shoot formation from primary shoots, which was more in presence of triacontanol (TRIA). Maximum number of shoots per culture was observed on the medium containing N6-benzyladenine (1.1 microM; BA), GA3 (5.8 microM) and PG (800 microM). Sub-culturing of the shoots onto MS medium containing optimum concentration of BA (5.6 microM), PG (200 microM) and TRIA (0.011 microM) produced elongated shoots along with secondary shoot formation. The long shoots were rooted on alpha-naphthalene acetic acid (5.38 microM; NAA) and PG (400 microM) containing medium. The rooted plantlets were hardened and their field survival rate was 80-90%.     

In vitro clonal multiplication of an apple rootstock by culture of shoot apices and axillary buds

In vitro clonal multiplication of apple rootstock MM 111 using axillary buds and shoot apices were carried out. Vegetative axillary buds of the size of 0.2-2.0 cm and shoot apices measuring 4 mm in length were initiated to shoot proliferation on MS medium supplemented with BA (0.5 - 1.0 mgl(-1)), GA3(0.5 mgl(-1)), with or without IBA(0.05 - 0.1 mgl(-1)). Small size explants showed less phenol exudation and less contamination. Following establishment phase, the small shoots emerged from explants were subcultured on MS medium supplemented with different combinations and concentrations of growth regulators. BA (1.0 mgl(-1)) and GA3 (0.5 mgl(-1)) combination showed highest multiplication rate (1:5), andcl also produced longer shoots. Two step rooting was done by transferring microcuttings to auxin free solid medium after root initiation in dark on 1/2 strength MS liquid medium containing IBA (0.5 mgl(-1) ). Rooted plantlets were transferred to peat containing paper cups and resulting plants of MM 111 acclimated successfully for transfer to field.     

Micropropagation ofFagus sylvatica L.

Summary An in vitro shoot multiplication system was established from juvenileFagus sylvatica L. tissues, and plantlets were regenerated. Embryonic axes were excised from beech seeds and germinated in vitro on media supplemented with 6-benzyladenine (BA) to obtain plantlets with axillary shoots. Shoot multiplication was maintained by sequential subculture of axillary shoot tips and basal segments on Woody Plant Medium supplemented with 0.5 mg/liter BA+2 mg/liter zeatin+0.2 mg/liter naphthaleneacetic acid (NAA). The effeciency of shoot multiplication clearly depended on the kind of explant used. Transfer to fresh medium every 2 wk during the 6-wk multiplication cycle improved multiplication rates. In the rooting stage, an initial 7-day dark period significantly improved rooting capacity and accelerated the emergence of roots on auxin-treated shoots. Adventitious buds were induced on the intact hypocotyls of the whole plantlets derived from the initial embryonic axis explants, especially on those cultured on medium with 1 mg/liter BA. Cotyledon and hypocotyl segments isolated from seedlings grown in vitro from embryos also exhibited capacity for adventitious bud formation, especially when cultured on media supplemented with 0.5 mg/liter BA + 0.1 mg/liter NAA.     

影响辣椒离体再生芽伸长因素研究

以辣椒子叶为外植体,比较不同浓度BA和IAA激素组合对辣椒再生芽诱导的差异,利用筛选出的高效芽诱导培养基为基础,研究了赤霉素、芽诱导时间、培养基有机成分、不同激素组合和品种等因素对辣椒不定芽伸长的影响。结果表明:不同基因型辣椒子叶再生能力不同,BA3.0mg.L-1 IAA0.5mg.L-1的激素配比对不定芽诱导频率最高;不定芽的伸长百分率随着GA3浓度的增加而增加,GA3的适合浓度为1.0～2.0mg.L-1;不定芽诱导时间对不定芽的伸长有一定的影响,诱导21d的不定芽,其伸长频率明显高于诱导14d的不定芽;B5有机成分在辣椒不定芽的伸长中效果优于MS有机成分;激素组合对不定芽伸长有一定的影响,Zeatin GA3激素组合对伸长效果最好,BA IAA GA3伸长效果较好,BA PAA(苯乙酸,phenylaceticacid) GA3伸长效果次之;不同品种辣椒不定芽的伸长能力有一定差异,楚风和苏椒五号再生芽伸长能力最佳。与IAA和NAA相比,IBA对再生芽生根效果较好。     

Micropropagation of meadowfoam (Limnanthes spp.)

Summary A rapid micropropagation system was developed for meadowfoam (Limnanthes spp. Brown) using four genotypes of three species. Murashige and Skoog (MS) medium supplemented with N⁶ benzyladenine (BA) and indole-3-acetic acid (IAA) at 0, 0.1, 0.5, 1.0 and 2.0 mg/l was tested for multiplication, shoot elongation and rooting. Expiants were taken from pot-grown plants. The most useful level for shoot growth and multiplication of both floral induced and non-induced plants was 0.5 mg/l BA. IAA failed to affect shoot growth or multiplication. Expiants from non-induced plants multiplied at moderate to high rates on 0.5 mg/l BA, while those from induced plants multiplied slowly and tended to elongate and flower. Non-induced plants on 2 mg/l BA produced large numbers of tiny shoots; induced plants did not respond. Shoots of all genotypes rooted on MS medium without hormones and all plants grew normally after transplanting to soil. This system provides a new tool for the development of meadowfoam as a crop plant.Abbreviations (BA) N ⁶ -benzyladenine - (IAA) indole-3-acetic acid - (MS) Murashige and Skoog medium, 1962     

Factors affecting recurrent shoot multiplication in in vitro cultures of 17- to 20-year-old douglas fir trees

Summary The effects of sucrose concentration, addition of ammonium nitrate, and exposure to N⁶-benzyl-adenine (BA) on multiplication potential with shoots derived from shoot cultures of 17- to 20-yr-old Douglas fir trees [Pseudotsuga menziesii (Mirb.) Franco] were compared. Each of these conditions, when compared independently, affected recurrent shoot multiplication and influenced shoot development, as measured by the abundance of shoot apices. Sucrose concentration was influential, the use of 25 g · liter⁻¹ providing twice the multiplication obtained with 20 g · liter⁻¹, and 14 × that obtained with the 30 g · liter⁻¹ concentration routinely used (tree 11). Ammonium nitrate usage also improved multiplication, a 2.5 times improvement being obtained after incorporation of 100 mg · liter⁻¹ NH₄NO₃ into the medium (tree 33). Shoot cultures were responsive but relatively sensitive to addition of BA, the best improvement in multiplication (5 times) being obtained with brief exposures to 3 mg · liter⁻¹ BA (tree 11). Although shoot cultures were responsive to the conditions investigated, differences in shoot multiplication and development were not displayed for several weeks. It was not possible therefore to repeat all the treatments with more than one genotype; however, when this was possible a genotype-dependent variation in response was evident.     

An efficient in vitro method for mass propagation of a woody ornamentalIxora coccinea L.

Various factors that affect culture establishment, shoot growth, proliferation and rooting ofIxora coccinea L., a woody shrub, were studied. Stem cuttings (decapitated shoot, three nodes) were the most suitable explants for multiple-shoot proliferation, and when cultured on a woody plant medium (WPM) containing 2.5 M BA produced axillary shoots which branched repeatedly, yielding an average of 27 shoots per explant after 6 weeks in culture. Kinetin, 2-iP, zeatin and thidiazuron all induced multiple-shoot formation, but were less effective than BA. While the presence of IAA in the multiplication medium was detrimental to shoot proliferation, shoot growth was not affected by IAA. The production of large amounts of basal callus and vitrification of shoots were the major problems to be avoided in proliferating shoot cultures. Addition of TIBA to the multiplication medium markedly reduced basal callusing, while sealing the culture vessels with a fluorocarbon polymer (tetrafluoroethyleneperfluoroalkyl vinyl ether) film (Neoflon PFA film) almost completely eliminated vitrification. A reduction in the number of vitrified shoots was also achieved with AVG treatment. Following this protocol of using BA-supplemented WPM and Neoflon film, it would be possible to produce more than 100,000 plants from a single stem cutting in 1 year.Abbreviations AVG Aminoethoxyvinylglycine - BA N⁶-benzyladenine - BM basal medium - IAA indole-3-acetic acid - IBA indole-3-butyric acid - 2-tiP N⁶-(2-isopentenyl)adenine - KIN kinetin - MS Murashige and Skoog medium - NAA -naphthaleneacetic acid - SRM shoot regeneration medium - TDZ thidiazuron - TIBA 2,3,5-triiodobenzoic acid - WPM woody plant medium - ZEA zeatin     

An efficient protocol for micropropagation of lemon (<Emphasis Type="Italic">Citrus limon</Emphasis>) from mature nodal segments

The influence of the basal medium and different plant growth regulators on micropropagation of nodal explants from mature trees of lemon cultivars was investigated. Although the basal medium did not affect any of the variables, explants on DKW medium were greener. Several combinations of 6-benzyladenine (BA) and gibberellic acid (GA) were used to optimise the proliferation phase. The number of shoots was dependent on the BA and GA concentrations and the best results were obtained with 2 mg l⁻¹ BA and 1 or 2 mg l⁻¹ GA. Explants length was shorter with the higher BA concentrations and, in all genotypes, shoot length was greater with 2 mg l⁻¹ GA. The best results for productivity (number of shoots × the average shoot length) were obtained with 2 mg l⁻¹ BA and 2 mg l⁻¹ GA, although explants with chlorosis and narrow leaves were observed. The presence of BA and GA in the proliferation medium was essential for the explant multiplication but GA had a greater influence. The transfer of in vitro shoots to rooting media, containing different concentrations of indole butyric acid (IBA) and indole acetic acid (IAA) produced complete plantlets. Lemon shoots rooted well in all rooting combinations. The highest rooting percentages were obtained on media containing 3 mg l⁻¹ IBA alone or IBA in combination with 1 mg l⁻¹ IAA and on these media the highest numbers of roots were produced. The average root length was affected significantly by the IBA and IAA concentrations. Root length was greater when only 3 mg l⁻¹ IBA was used, and in this rooting medium explants had a better appearance, with greener and larger leaves. The success during the acclimatisation was close to 100% and the plantlets exhibited normal growth in soil under greenhouse conditions.     

Some Physiological Characteristics of Two Tomato Cultivars, One Tolerant and One Susceptible to Tobacco Mosaic Virus

A comparison was made of physiological activity in the tomatocv. Potentate, susceptible to tobacco mosaic virus (TMV) withthat in cv. Virocross which is tolerant. Growth analysis overa 5-week period showed Potentate to have a higher relative growthrate and a higher net assimilation rate than Virocross. In reciprocalgrafts scion growth, including dry weight, was greater on Potentaterootstocks than on Virocross. Growth responses were studiedin these grafts following foliar sprays of N-6 benzyladenine(BA) and gibberellic acid (GA). The response of the scion toGA was similar in both cultivars and there was no interactionbetween GA and rootstock for scion growth. An interaction betweenstock and BA was found for leaf growth. Bleeding sap from Virocross contained more gibberellins, morecytokinins, and a lower concentration of amino compounds, especiallynon-protein acids, than Potentate. There were more gibberellinsin the roots of Potentate but no difference between shoots ofthe two cultivars. In reciprocal grafts TMV multiplied more rapidly in Virocrossscions on Potentate rootstock than on their own, but Virocrossstocks did not affect virus multiplication in Potentate scions.Spraying Potentate plants with BA reduced the rate of virusmultiplication. It is suggested that virus multiplication in the shoot may becontrolled by two distinct root factors, viz, the supply ofnative cytokinins and the amount and nature of the nitrogenouscompounds which directly influence the rate of shoot growth.