盐节木同化枝的腋芽丛生与快速繁殖

本文通过同化枝腋芽丛生方式建立了盐生植物盐节木的快速繁殖体系。以在MS附加200 mmol·L-1 Na Cl的培养基上无菌种子萌发、生长共计4~5个月、高约3~4 cm的盐节木小苗为材料,从上截取0.5 cm长的同化枝小段为外植体,培养在MS附加不同浓度的6-BA与NAA的培养基中。结果表明:同化枝腋芽诱导与增殖最佳培养基均为MS+0.05 mg·L-1 6-BA+1mg·L-1 NAA或MS+1 mg·L-1 6-BA,初代培养30 d后,腋芽诱导率达97.5%,平均芽增殖系数为2.50;继代增殖培养时,将同化枝切成1 cm长的小段芽增殖系数提高到4.92;一次性芽苗伸长与生根培养基为MS+0.05 mg·L-1 IBA,生根率达99%;小苗经炼苗后移栽到沙子、营养土和蛭石体积比为1:1:1的基质中,成活率可达91%。     

马来沉香组织培养技术研究

以马来沉香茎段为外植体,分别对外植体的消毒、启动培养、增殖培养、壮苗培养、生根培养、炼苗移栽环节进行研究,着重探索马来沉香组织培养技术各个环节的最佳培养基配方,为马来沉香的工厂化育苗提供技术指导。结果表明:马来沉香最佳消毒方法是用0.1%升汞消毒4～5min;启动率最高的培养基配方是1/2MS+6-BA 0.2mg·L-1+NAA 0.1mg·L-1+蔗糖30g·L-1+琼脂5.8g·L-1,启动率达70.5%;增殖系数最高的培养基是1/2MS+0.1mg·L-16-BA+25g·L-1蔗糖+5.8g·L-1琼脂,增殖系数达2.9;最佳壮苗培养基是1/2MS+30g·L-1蔗糖+5.8g·L-1琼脂;最佳生根培养基为1/2MS+NAA 5.0mg·L-1+20g·L-1糖+6g·L-1琼脂,培养2d后移入1/2MS培养基继续培养,生根率为83%;马来沉香移栽较难成活,在泥炭土∶黄泥土(2∶1)的基质上成活率最高,移栽成活率65%。     

黑莓品种‘Chester’离体培养过程中适宜激素种类和浓度以及光照度的筛选

以黑莓(Rubus spp. )品种'Chester'带腋芽茎段为外植体,对初代培养基、不定芽增殖培养基和生根培养基中适宜的激素种类及浓度进行了筛选,并对增殖培养过程中适宜的光照度进行了研究.结果显示,在初代培养基中添加不同质量浓度的6-BA和NAA对侧芽的萌发和生长有明显的影响,其中,NAA不利于芽的萌发和生长,而添加适量的6-BA可促进芽的萌发和生长;适宜于'Chester'茎段的初代培养基为添加了1.00 mg·L-16-BA的MS培养基(含25 g·L-1蔗糖和5.9 g·L-1琼脂,pH 5.8).单因素实验结果显示,在 'Chester'不定芽的增殖过程中,细胞分裂素主要影响不定芽增殖,而生长素主要影响不定芽生长,以质量浓度低于1.0 mg·L-1的6-BA以及质量浓度低于0.3 mg·L-1的NAA较为适宜;经过进一步的组合筛选,确定适宜于'Chester'不定芽增殖的培养基为添加了0.5 mg·L-16-BA和0.1 mg·L-1NAA的MS培养基(含25 g·L-1蔗糖和5.9 g·L-1琼脂,pH 5.8).在生根培养基中添加高浓度NAA易诱导不定芽基部产生愈伤组织,不利于不定芽生根,因而,'Chester'不定芽适宜的生根培养基为添加了0.10 mg·L-1NAA的1/2MS培养基(含20 g·L-1蔗糖和5.9 g·L-1琼脂,pH 5.8).在不定芽的增殖培养过程中,光照度较强有利于不定芽生长,适宜的增殖培养条件为:光照度3 000 lx、光照时间15 h·d-1、温度(25±2) ℃、相对湿度约70%,此条件下不定芽的增殖系数(1.95)和平均芽长(1.44 cm)最高,芽生长状况良好.     

葡萄柚种子无菌苗组培快繁体系的建立

以葡萄柚种子在无菌条件下萌发的幼苗作为外植体供体,子叶切断作为丛生芽初代诱导培养的接种材料,以MT作为基本培养基,通过调节不同植物生长调节剂及其浓度组合、不同质量浓度的蔗糖,选择最佳初代培养基、继代培养基和生根培养基,对葡萄柚组培快繁技术体系进行研究。结果表明:(1)葡萄柚种子经预处理后,先用75%酒精表面灭菌15 s,再用0.1%Hg Cl2浸泡20 min的消毒效果最好,污染率为18.33%,萌发率为89.91%;(2)初代培养的最适培养基为MT+1.0 mg·L-16-BA(6-苄氨基腺嘌呤)+0.2 mg·L-1IBA(吲哚丁酸)+0.2 mg·L-1GA(赤霉素)+蔗糖40 g·L-1,腋芽诱导率较高,为93.33%;(3)浓度为0.2 mg·L-1的GA能提高初代培养的腋芽诱导率,但还达不到显著水平;(4)继代培养的最适培养基为MT+0.5 mg·L-16-BA+0.2 mg·L-1IBA+0.2 mg·L-1GA+蔗糖40 g·L-1,丛生芽增殖系数达到4.25,芽深绿色,茎粗壮,节间较长,生长旺盛;(5)浓度为0.2 mg·L-1的GA能显著提高继代培养的增殖系数;(6)最适合继代培养的蔗糖浓度为40 g·L-1,丛生芽长势极好;(7)最佳生根培养基为1/2 MT+NAA 0.2 mg/L+蔗糖40 g·L-1+AC 0.1%,生根率达68.89%,根粗壮;(8)生根苗移栽30 d后成活率在75%以上。该研究建立了葡萄柚的组织培养快繁技术体系,为葡萄柚的规模化生产提供了可行的技术依据。     

濒危植物盘龙参种子的非共生萌发及种苗的快繁研究

以盘龙参种子为材料,筛选离体条件下适宜种子非共生萌发、种苗快繁的培养基。结果表明:盘龙参种子在无植物生长物质的培养基中能够萌发,但不能发育成苗;在含有1.0mg·L-1 KT、0.1mg·L-1 IAA和0.1mg·L-1 GA3的培养基中萌发,并能进一步发育形成种苗;在较高浓度生长素(1.2mg·L-1 NAA)的培养基中不能萌发。种苗在较高浓度细胞分裂素和较低浓度生长素配比的培养基中能够增殖,最高增殖系数可达到2.8,转入壮苗生长培养基中培养80d后可以移栽温室。最适增殖培养基为1/2MS+12.0mg·L-1 6-BA+0.1mg·L-1 NAA+10.0mg·L-1腺嘌呤;最适壮苗生根培养基为1/2MS+1.0mg·L-1 KT+0.1mg·L-1 IAA+10.0mg·L-1腺嘌呤。     

烤烟‘K326’的组织培养与快速繁殖

为了提供市场所需的大量种苗,适应云南烤烟(Nicotiana tabacum)规模化生产的需要,以田间烤烟的叶片、茎段、培养瓶内种子萌发的叶片和茎段为外植体进行组织培养实验.实验比较了5种培养基,筛选出烤烟的最佳愈伤组织诱导和愈伤组织不定芽分化培养基为MS 6-BA 2 mg·L-1 NAA0.2 mg·L-1 Sugar30 g·L-1 Agar6 g·L-1;生根培养基为MS Sugar30g·L-1 Agar6 g·L-1,生根率为100%;用营养土移栽后,存活率为100%.     

烤烟‘K326’的组织培养与快速繁殖

为了提供市场所需的大量种苗,适应云南烤烟(Nicotiana tabacum)规模化生产的需要,以田间烤烟的叶片、茎段、培养瓶内种子萌发的叶片和茎段为外植体进行组织培养实验。实验比较了5种培养基,筛选出烤烟的最佳愈伤组织诱导和愈伤组织不定芽分化培养基为MS+6-BA 2 mg.L-1+NAA 0.2 mg.L-1+Sugar 30 g.L-1+Agar 6 g.L-1; 生根培养基为MS+Sugar 30 g.L-1+Agar 6 g.L-1,生根率为100%;用营养土移栽后,存活率为100%。     

多花兰种子无菌萌发及离体快繁研究

以多花兰种子为材料,研究了无机盐浓度、植物生长调节剂和光照条件对多花兰种子非共生萌发的影响,在此基础上,通过研究原球茎增殖和分化、芽苗壮苗和生根的培养基配方及培养条件,建立多花兰组培快繁技术体系.结果表明:多花兰种子萌发培养基为1/6 MS十NAA0.5 mg·L-1+6-BA 2.0 mg· L-1+马铃薯泥50g·L-1+AC 1.0g·L-1,光照度为1.25μmol·m-2·s-1,萌发率63.6％;原球茎增殖继代培养基为1/4 MS+6-BA2.0 mg·L-1+NAA 0.5 mg· L1+AC 1 g·L-1+PE 200 g·L-1,繁殖倍数6.5倍/60 d,芽分化率60.2％;再生芽分化培养基为1/4 MS十6-BA2.0mg·L-1+NAA 0.2 mg· L-1+AC 1 g·L-1+PE 200g· L-1,繁殖倍数4.0倍/60 d,芽分化率85.0％;芽苗壮苗和生根培养基为1/6 MS+6-BA 3.0 mg·L-1+NAA 1.0 mg·L-1+AC 1 g·L-1+蔗糖20g·L-1 +PE 200 g·L-1和1/4 MS+6-BA 2.0 mg·L-1+NAA1.2 mg·L-1+AC1 g·L-1十蔗糖20g·L-1+PE200g· L-1,生根率达100％,生根苗移栽成活率90％.此技术可用于多花兰种苗繁育和种质资源保护.     

石龙尾(Limnophila sessiliflora BI.)的组织培养与快速繁殖技术研究

以石龙尾(Limnophila sessiliflora BI.)沉水枝带节茎段为外植体进行离体培养,研究外植体灭菌方法以及培养基中不同生长调节剂的浓度对其增殖、生根的影响。结果表明:以0.1%的HgCl2为灭菌剂,采用4 min+4 min、间歇4 h的间歇灭菌法,可以获得成活的无菌外植体15%;在1/2 MS+6-BA 2.0 mg·L^-1+NAA 0.1~0.2 mg·L^-1的增殖培养基上培养35 d,试管苗的增殖系数可达30.8以上;在1/2 MS+6-BA 0.3 mg·L^-1+NAA0.5 mg·L^-1的生根培养基上培养28 d后,可获得具3~5个侧枝的生根苗,平均每株生根数4.8条;炼苗后移植成活率100%。     

新塔花的组织培养与快速繁殖

以新疆特有药用植物新塔花(Ziziphora bungeana Juz.)幼嫩茎段为外植体材料,筛选初代培养、继代增殖与生根培养的条件,建立了新塔花组织培养与快速繁殖体系。结果表明:新塔花种子最适萌发培养基为MS+0.25 mg·L~(-1) NAA,萌发率达57.78%;茎段诱导腋芽最适培养基为MS+0.5 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA,诱导率为90.47%;适宜芽增殖培养基为MS+0.5 mg·L~(-1 )6-BA+0.1 mg·L~(-1) NAA+0.3 mg·L~(-1) GA_3+35 g·L~(-1)蔗糖,20 d增殖系数达1.95;最佳诱导根生长培养基为1/2MS+0.5 mg·L~(-1) NAA+0.2%AC,生根率达60%,平均生根数为3.5,20 d株高达4.5 cm。移栽至混合基质(营养土:珍珠岩:蛭石=1:1:1)中,组培苗成活率为80.45%。     

软枣猕猴桃‘天源红''离体再生体系的建立

为了建立快速高效的全红型软枣猕猴桃离体再生体系,该研究以果皮、果肉均为红色的软枣猕猴桃新品种‘天源红’( Actinidia arguta)带腋芽茎段和幼嫩叶片、叶柄为外植体材料,采用组织培养的方法,研究适合其离体再生的外植体类型以及最佳植物生长物质组合。结果表明：初春带腋芽的茎段是最好的获得无菌苗的外植体材料,诱导腋芽出芽的最佳植物生长物质组合为MS＋6－BA 0.5 mg·L-1＋IBA 1.0 mg·L-1;研究发现叶柄比叶片更适合进行‘天源红’愈伤组织诱导,叶柄诱导的最佳植物生长物质组合为MS＋ZT 0.5 mg·L-1;同时,研究了不定芽增殖的最佳植物生长物质组合为MS＋ZT 1.0 mg·L-1;此外使用6－BA也可以达到较高的不定芽增殖率,在生产上可以替代ZT进行不定芽分化,即MS＋6－BA 2.0 mg·L-1＋IBA 0.5 mg·L-1;较适宜的生根培养基为1/2 MS＋NAA 0.2 mg·L-1;生根后的组培苗在珍珠岩∶泥炭∶细沙＝1∶1∶1的基质配比中能够达到98％的移栽成活率。该研究结果建立了全红型软枣猕猴桃的离体再生体系,为全红型软枣猕猴桃苗木快繁、工厂化育苗提供了技术支持,同时建立的再生体系为软枣猕猴桃遗传转化研究提供了基础。     

红厚壳茎段丛生芽诱导与植株再生

红厚壳(Calophyllum inophyllum)为藤黄科红厚壳属多年生木本植物,有很高的药用价值。该研究以红厚壳带节茎段为外植体,探讨生长调节剂对腋芽萌发及丛生芽诱导、伸长和试管苗生根的影响。研究结果表明,外植体腋芽萌发和丛生芽诱导效果最好的培养基是MS+NAA1.0+TDZ0.5,在此条件下培养21天后,转入添加0.5 g·L–1活性炭且无生长调节剂的MS培养基,可有效促进不定芽的伸长。将带不定芽的外植体先在附加1.0 mg·L–1NAA的1/2MS培养基上进行生根诱导4周,之后转入附加1.0 g·L–1活性炭的无激素培养基进行根的伸长培养,这样的两步生根法能有效促进红厚壳生根。     

梨枣叶片和茎段再生体系的建立

用不同浓度配比的生长素和细胞分裂素诱导梨枣叶片和茎段愈伤组织的产生,并研究了不定芽诱导的最佳配方,建立了梨枣叶片和茎段的再生体系.结果表明,梨枣叶片愈伤组织诱导的最佳培养基为MS+2,4-D 1.5 mg·L-1+6-BA 0.5 mg·L-1;茎段为MS+2,4-D 1.0 mg·L-1+6-BA 0.5 mg·L-1.叶片不定芽诱导的最佳培养基为MS+IBA 0.1 mg·L-1+6-BA 1.5 mg·L-1. AgNO3能阻止叶片外植体褐化并有效地促进叶片愈伤组织分化.茎段能在同一培养基上产生愈伤组织并直接分化出不定芽.     

小叶龙竹（Dendrocalamus barbatus）愈伤组织诱导及植株再生

以小叶龙竹种子为外植体,通过研究MS培养基中不同植物生长调节剂浓度组合对外植体愈伤组织诱导和不定芽分化的影响以及不同配比对生根的作用,建立了稳定的繁殖再生体系。试验结果表明愈伤组织诱导的最适培养基为MS＋2,4-D5．0mg·L-1;不定芽分化的最适培养基为MS＋2,4-D0．5mg·L-1＋6．BA1．0mg·L-1＋KT0．25mg·L-1;小苗的最适生根培养基为1／2MS＋6-BA0．5mg·L-1＋NAA1．0mg·L-1＋IBA0．4mg·L-1,建立的繁殖再生体系将为进一步利用分子生物学技术对竹类植物进行遗传改良奠定基础。     

白桦成熟合子胚快速不定芽诱导体系的建立

以白桦成熟合子胚为外植体,建,3-7白桦合子胚快速不定芽诱导体系：合子胚经消毒纵切后,在培养基WPM＋2．0mg·L-16-BA＋0．2mg·L-1NAA中,可直接诱导形成不定芽,诱导率最高为93．3％,不定芽诱导数为8．7个;WPM＋4．0mg·L-1TDZ上也有较好的诱导效果,不定芽诱导率为90．2％,不定芽诱导数为7．6个,这表明WPM＋2．0mg·L-1 6-BA＋0．2mg·L-1 NAA是白桦成熟合子胚不定芽诱导的较优培养基。NAA能有效促进嫩茎生根,其最佳浓度为0．2mg·L-1。     

多花黄精诱导愈伤组织与不定芽培养基筛选

以多花黄精Polygonatum cyrtonema的根状茎为外植体,通过L9(34)正交试验比较不同植物生长调节剂及其组合对多花黄精愈伤组织诱导的影响,筛选最适生长调节剂配方,同时筛选通过器官发生方式直接成芽较为合适的培养基。结果表明,含有2,4-D和KT的培养基诱导愈伤组织效果显著,多花黄精愈伤组织诱导的最适培养基为MS + 6-BA 2.0 mg·L-1 + 2,4-D 0.5 mg·L-1 + NAA 0.1 mg·L-1 + KT 1.0 mg·L-1,该培养基的愈伤组织诱导率可达39.10%;培养基MS + 6-BA 4.0 mg·L-1 + NAA 0.2 mg·L-1可诱导多花黄精根状茎直接产生不定芽。     

Rapid in vitro propagation of Aloe barbadensis Mill

Axillary bud development and adventitious bud formation was obtained with decapitated shoot explants of Aloe barbadensis Mill. Maximal bud growth and rooting of shoots was obtained on a modified medium of Murashige and Skoog supplemented with 5 M IBA. More adventitious and axillary buds developed on nutrient media supplemented with IBA than with NAA. Axillary buds but not adventitious buds developed with IAA in the medium. Morphogenesis was inhibited by 2,4-D. Kinetin, benzyladenine and thidiazuron were toxic to the explants and did not stimulate the development of axillary of adventitious buds. The optimal temperature for bud growth and development was 25°C. Axillary bud growth and the formation of adventitious buds was slowed down at 10°C and totally inhibited by 30°C. The optimal sucrose concentration was 3% with the inhibition of bud growth and development by higher sucrose levels.     

红籽鸢尾高频离体再生条件的初步筛选

红籽鸢尾(Iris foetidissima Linn.)为鸢尾科(Iridaceae)鸢尾属(Iris Linn.)多年生草本植物,原产于西欧和非洲北部。该种的叶鞘呈深绿色剑形,蒴果饱满且蒴果成熟开裂后露出红、橙黄和白等不同色泽的种子并一直垂挂到翌年春天,集观花、观叶和观果为一体,观赏价值较高。另外,该种还具有耐寒性好、在长江中下游地区四季常绿、适应性强及耐粗放管理等优     

In vitro differentiation and plant regeneration of Albizia chinesis (Osb.) Merr

Summary Petiolar and distal cotyledonary segments (PCS and DCS) of Albizia chinensis were cultured on Murashige and Skoog's (MS; 1962) medium and induced to form adventitious shoot buds in the presence of either cytokinins 6-benzylamino purine (BAP), kinetin (KN) or thidiazuron (TDZ). Superiority of BAP in inducing shoot bud and differentiation was observed. PCS was more morphogenic to shoot bud differentiation than DCS. TDZ was highly effective in inducing shoot buds, but arrested shoot growth, while KN produced more callus during differentiation of shoots. Rapid and high rate of shoot multiplication per explant was achieved through subculture in MS medium containing BAP (1.0 mg l⁻¹) and indole-3-acetic acid (IAA) (0.5 mg l⁻¹). BAP at low concentration was required to enhance shoot multiplication and elongation. Successful rooting of regenerated shoots was carried out in a two-step culture procedure in MS media with indole-3-butyric acid (IBA) (2.0 mg l⁻¹) and subsequent subculture in IBA-free medium.     

Floral-stimulus activity in tobacco stem pieces

The growth patterns of axillary buds of dayneutral tobacco (Nicotiana tabacum L. cv. Wisconsin 38) plants were assessed by using expiants of single buds attached to leafless stem cuttings and allowing the buds to grow to flowering without additional manipulation. Buds located 5, 10 and 15 nodes below the inflorescence were employed. For a given bud position, when a cutting had few internodes the growth pattern of a bud tended to fall into one of two groups: buds that produced few-noded shoots and buds that produced many-noded shoots. For example, in a group of 13 cuttings composed of bud 5 with 2 associated internodes, 11 buds produced 14.2 nodes (range, 11–17) and 2 buds produced 32.0 nodes (range, 30–34). As the number of internodes on the cutting increased, the number of buds producing few-noded shoots increased and the number of nodes produced decreased (e.g. in contrast to the data above, all 5th buds with 6 internodes produced 12.8 nodes; range 11–15). When cuttings from the 3 positions had the same number of internodes, the more apical cuttings had buds that produced fewer nodes (e.g. for cuttings with 6 internodes all 5th buds produced 12.8 nodes, all 10th buds produced 15.5 nodes and 85% of 15th buds produced few-noded shoots with 19.3 nodes). The number of nodes produced by a bud was a function of the original position of the stem piece and not the original position of the bud. That is, bud 5 associated with the 6 internodes below it produced 12.8 nodes and bud 10 associated with essentially the same 6 internodes (i.e. the 6 above it) produced 12.9 nodes while bud 10 associated with the 6 internodes below it produced 15.5 nodes. Thus, the number of nodes produced by a bud was dependent upon the original main-axis position of the cutting as well as the number of internodes on the cutting. Buds forced to grow out in situ on main axes devoid of leaves produced substantially more nodes than similar buds on cuttings. Buds isolated without associated internodes produced many-noded plants with a number of nodes similar to that of plants grown from seed. The simplest interpretation of these data is that stem pieces contain floral-stimulus activity and that this activity is present in a gradient with the highest activity being located in the apical part of the stem.We thank Susan Smith and Harry Roy (Rensselaer) for comments, and the National Science Foundation for financial support (IBN-9003739 to C.N.M.).