草麻黄植株再生体系的建立

目的:以草麻黄的子叶为材料,建立草麻黄植株再生体系.方法:采用组织培养的方法进行了愈伤组织诱导、愈伤组织分化和不定芽生根研究.结果:MS+2,4-D 2.0mg/1+6-BA 1.0mg/l为诱导子叶形成具有分化能力愈伤组织的理想培养基;MS+2,4-D 1.5mg/l+6-BA 1.5mg/l是愈伤组织的最佳继代培养基;MS+IAA 0.2mg/l+TDZ 2.0mg/l是愈伤组织不定芽分化的最佳培养基,分化率为75%;试管苗生根培养基为MS+2,4-D 1.0mg/l.结论:建立了草麻黄子叶再生系统,为开发和保护麻黄野生资源提供一定的材料来源和技术方法.     

红豆草体细胞胚胎发生的激素调控及L—脯氨酸对它的促进作用

在 LS 附加1mg/1 BA+1mg/l KT 的培养基上,红豆草(Onobrychis viciaefolia Scop.)无菌苗的下胚轴切段产生淡黄色的愈伤组织。愈伤组织转移到 LS 附加1mg/l BA 的培养基上,诱导体细胞胚胎发生,而在 LS 附加1mg/l KT 的培养基上抑制体细胞胚胎发生。同时,发现红豆草胚性愈伤组织中游离脯氨酸的含量仅为非胚性愈伤组织的2/5。向培养基中加入L-脯氨酸可以促进红豆草体细胞胚胎发生。最适浓度为1000mg/l。     

杜衡离体繁殖的研究

以马兜铃科植物杜衡为研究材料,试图通过茎尖培养来获得愈伤组织,继而诱导愈伤组织分化形成杜衡小植株。试验证明：杜衡茎尖在MS基本培养基附加6-BA 0.6mg/l和NAA0.1mg/;这一激素组合上可诱导茎尖基部形成愈伤组织;将愈伤组织分割转接到附加6-BA0.2mg/l和NAA0.01mg/l 的MS基本幅度基上可使愈伤组织分化形成小芽;分化形成的芽置于1/8-1/4MS(大量元素减少,其余成分不变)附加6-BA0.1mg/l和GA1mg/l的培养基上,可促使芽的正常生长;新生芽转接在1/4MS附加IBA0.5mg/l培养基上促使其生根。     

栀子组织和细胞培养生产天然食用色素的研究——Ⅰ.愈伤组织培养的研究

在诱导出愈伤组织的基础上,对各种不同的培养条件进行分析研究,考察它们对愈伤组织生长和栀子黄色素产生的影响,筛选出适宜的生长培养基:B_5+IBA 1mg/l+KT0.23mg/l、MG-5+IBA 1mg/l+KT 0.23mg/l、B_6+IAA 1.5mg/l和生产培养基:M-9+IAA 1mg/l、M-9+IAA 1.5mg/l.并且,获得了几个色素含量较高的愈伤组织系。另外,还研究了含色素和不含色素的愈伤组织在培养过程中的过氧化物同工酶的差异。     

防风悬浮细胞的原生质体再生植株

防风(Saposhnikovia divaricata(Turcz.)Schischk)试管苗的根尖,下胚轴或叶柄切段在含有1mg/l 2,4-D 的 MS 固体培养基上,形成含有胚性细胞团的愈伤组织。愈伤组织经液体振荡培养,形成含有大量胚性细胞团的悬浮培养物。用含有 Onozuka R-10 1.5%、Mace-rozyme R-10 0.3%、蜗牛酶0.5%、CaCl_2 5mmol/l 和甘露醇0.6 mol/l(pH=5.8)的酶液从胚性细胞团游离得到原生质体。原生质体在培养的第4天出现第一次分裂,50天左右形成的细胞团大小为1—2mm。这些细胞团在含有0.5 mg/l 2,4-D 的 MS 固体培养基上形成愈伤组织。在含有0.1 mg/l 6-BA 或0.1 mg/l 2,4-D+0.5mg/l 6-BA 的 MS 固体培养基上,原生质体再生的愈伤组织分化出胚状体。胚状体在不含任何生长调节剂的 MS 固体培养基上发育成完整的原生质体再生植株。     

白刺花胚性愈伤组织诱导及体细胞胚发生和萌发

探讨不同因素对白刺花下胚轴、子叶2种外植体胚性愈伤组织诱导及体细胞胚发生和萌发的影响。以B5和MS为基本培养基,研究2,4-D、6-BA和TDZ对白刺花下胚轴和子叶胚性愈伤组织的诱导;在MS培养基上添加不同浓度2,4-D,研究胚性愈伤组织增殖情况;采用ABA,探究对体细胞胚发生的影响。结果表明:下胚轴比子叶更易诱导胚性愈伤组织,筛选出2种外植最佳的胚性愈伤组织诱导培养基均为MS+2.0 mg/L 2,4-D+0.5 mg/L TDZ+0.5 mg/L 6-BA,胚性愈伤组织诱导率分别为77.3%和41.0%。15.0 mg/L ABA、0.2 mg/L 2,4-D和2.0 mg/L 6-BA有利于体细胞胚发生,1/3MS+0.2 mg/L NAA+0.1 mg/L 6-BA+2.0 g/L活性炭+25 g/L蔗糖+7 g/L琼脂的培养基可使体细胞胚萌发率达80%以上,再生植株移栽成活率高达90%。白刺花外植体种类及培养基类型均会影响胚性愈伤组织的诱导,其中下胚轴诱导效果优于子叶;MS培养基较适合启动细胞脱分化形成愈伤组织,2,4-D对胚性愈伤组织的增殖保持有调控作用,ABA有利于体细胞胚的发生。     

伊贝母(Fritillariae pallidiflorae)愈伤组织的诱导和植株再生的研究

本文对伊贝母鳞瓣切块和鳞芽顶端愈伤组织的形成和鳞茎的再生进行了研究。结果在加有2.4—D1毫克/升和KT0.1毫克/升的MS培养基上诱导出了愈伤组织。在加有IAA1毫克/升和NAA0.2毫克/升及KT0.1毫克/升的MS培养基上分化出了小鳞茎。培养结果表明:小鳞茎有三个来源:(1)由鳞瓣切块的愈伤组织形成。(2)由鳞瓣切块直接发育成。(3)由鳞芽顶端形成。所形成的小鳞茎和大田栽培的没有什么不同。培养4个月的小鳞茎相当于种子繁殖2—3年的鳞茎一样大小。     

绿花百合胚性愈伤组织诱导与植株再生研究

建立绿花百合(Lilium fargesii Franch.)胚性愈伤组织诱导体系,为保护和合理利用这一重要植物资源提供高效、稳定的再生技术途径。以绿花百合鳞片为外植体,通过正交实验研究不同激素种类及其质量浓度对绿花百合胚性愈伤组织诱导、胚状体和小鳞茎分化及植株再生的影响。结果表明:较适宜诱导胚性愈伤组织的培养基为MS+6-BA 0.5 mg·L~(-1)+NAA 0.5 mg·L~(-1)+2,4-D 0.1 mg·L~(-1),出愈率达89.29%,小鳞茎发生系数亦达4.7;胚性愈伤组织增殖及小鳞茎发生培养基为MS+6-BA 1.0 mg·L~(-1)+2,4-D 0.1 mg·L~(-1),繁殖倍数5.0/35 d;根的诱导则在1/2MS+NAA 0.2 mg·L~(-1)的培养基上进行,生根率达100%,幼苗移栽至排水良好的沙土中,保温保湿培养35 d后,成活率可达90%以上。本研究为保持绿花百合优良品种特性、种苗繁育提供了有效途径,也为保护其野生资源,发展人工栽培和利用胚状体进行无性育种奠定了基础。     

小麦×天兰冰草杂种体细胞愈伤组织诱导及无性系建立

从普通小麦×天兰冰草杂种F_1的叶、茎、节、幼穗诱导出愈伤组织,建立了体细胞无性系,获得了大量试管苗并移栽成活。 愈伤组织诱导率及分化率以幼穗最高,茎、节、叶较低。完全展开的叶片不能形成愈伤组织,未伸展的幼叶能诱导出愈伤组织,分化程度越低的幼叶部位越容易脱分化。幼叶基部切段的愈伤组织诱导率可达90％以上。改良MS附加4mg/l 2,4-D、0.1mg/l KT、0.5mg/l NAA为最佳愈伤组织诱导培养基。最适宜的分化培养基为改良MS附加0.25mg/l KT、0.5 mg/l NAA、150 mg/l腺嘌呤核苷。杂种的愈伤组织诱导率及分化率高于两个亲本。杂种的愈伤组织长势旺盛,适应性强等都表现了小冰麦杂种在组织培养中的杂种优势,而且这种再生能力杂种优势可以通过无性系得到保持。     

辣木资源利用中的悬浮细胞培养体系研究

辣木富含多种营养成分,在食品和药物开发方面有巨大的潜在开发价值。本文提供了一种可行的辣木细胞悬浮培养技术。由辣木的根诱导形成愈伤组织和叶诱导形成愈伤组织的合适细胞悬浮培养条件分别为MS培养基(MS)+1.0mg/L 2,4-二氯苯氧乙酸(2,4-D)+1.0mg/L激动素(KT)和MS+0.5mg/L 2,4-D+0.5mg/L KT,摇床转速均为50~100r/min,将愈伤组织添加到液体悬浮培养基中20d左右可得到大量悬浮细胞。本研究为辣木细胞水平的培养和研究提供了一条途径,为辣木潜在价值的开发利用提供新的思路。     

火百合花器的鳞茎诱导和增殖培养研究

采用4.5 ～6.5cm长、花苞青色未开放的火百合花蕾作材料,切取其子房、花柱、花丝为外植体进行鳞茎诱导,结果表明:花丝的鳞茎诱导率最高,其次是花柱,子房的诱导率最低。以子房和花柱为外植体诱导出的鳞茎个体大小不一(1～7mm) ,以花丝为外植体诱导出的鳞茎则普遍偏小,但较整齐一致(3～4mm) ;小鳞茎诱导的最佳6-BA浓度是1.0mg/L。研究了基本培养基、6-BA浓度、NAA浓度对小鳞茎增殖的效应及不同蔗糖浓度对小鳞茎增重的影响,结果表明:小鳞茎增殖的最佳培养基是:MS+6-BA2.0 mg/L+NAA0.2 mg/L,4 %蔗糖对鳞茎增重效果最好。     

Enhanced bud and bulblet regeneration from bulbs of<Emphasis Type="Italic"> Nerine sarniensis</Emphasis> cultured in vitro

Nerine (Nerine sarniensis) cv. Salmon Supreme in vitro-grown bulblets, 7-9 mm in diameter, were cut in half longitudinally and used for adventitious bud initiation following dissection of the roots and two-thirds of the upper part of the bulblets. The terminal apex was injured with a hot, sterile microscope dissecting needle. The highest number of buds formed (seven to nine buds per halved bulblet) on a semi-solid Murashige and Skoog (MS) basal salts medium supplemented with 3% sucrose and either 1 microM 6-benzylaminopurine (BA) and 1 microM alpha-naphthaleneacetic acid (NAA) or 0.5 microM BA and 0.1 microM NAA. Bulblet halves were cultured in the dark for 11-13 weeks with one subculture after 6 weeks. Anatomical studies indicated that the initiation of adventitious buds on the abaxial side of the inner scales of the halved bulblet was adjacent to the basal plate and started from the leaf primordium and a meristematic bulge. Buds developed directly into small bulblets after they were transferred to semi-solid MS basal salts medium supplemented with 6% sucrose, 10 microM indole-3-butyric acid (IBA) and 0.25% activated charcoal. Small bulblets cultured in liquid MS medium supplemented with additional KH(2)PO(4 )(170 mg l(-1)), 6% sucrose and 0.1 microM NAA under a 16/8-h (light/dark) photoperiod for 8 weeks grew into larger bulbs faster than those cultured on semi-solid medium. The bulbs were rooted on a semi-solid medium after 4 weeks and then transferred to the soil. As many as 18 bulblets developed and rooted from one in vitro-grown bulb after 25-27 weeks.     

In vitro bulblet production of Lachenalia

In vitro bulblet formation and subsequent transplanting of bulblets to soil were studied in order to develop a cost-effective method for the mass production of three Lachenalia varieties. Clumps of adventitious shoots regenerated from leaf explants were used. Bulblet formation was initiated after 2 weeks when shoots were subjected to low temperature (4–15 °C). The size (age) of the adventitious shoot affected the bulblet size, and shoots shorter than 4 mm did not form bulblets. Larger bulblets formed on medium containing 6% sucrose compared to 3% sucrose. Following bulblet initiation, illumination was not necessary for the completion of bulblet formation. Bulblets went into dormancy 3–4 months after they had been initiated or when the culture medium dried out, and they were released from dormancy when the natural night temperatures started to decrease in the late summer. The survival rate of the bulblets after transplanting was directly correlated to the size of the bulblets.The most important factors influencing in vitro bulblet formation of Lachenalia were sucrose concentration, temperature and length of explant shoots. Received: 12 June 1998 / Revision received: 8 September 1998 / Accepted: 23 September 1998     

Differentiation in Lilium Bulbscales Grown in vitro. Effect of Various Cultural Conditions

Tissue cultures of Lilium auratum Lindl. and L. speciosum Thunb., which were derived from bulbscales, all appeared to differentiate organs. The effect of cultural conditions on the differentiation of bulblets and roots was examined. The best material for bulblet formation was bulbscales of intact or in vitro produced bulblets. The optimum temperature was 20°C and optimum pH was 6. Effect of irradiance on organ formation was not obvious but leaf emergence was stimulated. Higher kinetin concentrations stimulate the formation of numerous bulbscalcs. High NAA concentrations induce roots. On the other hand kinetin inhibits the NAA effect on root formation. A high sucrose concentration stimulated organ formation, but the number of bulblets was at a constant level in the medium containing between 10 and 90 g/l of sucrose. The formation of bulblets and their growth were stimulated at increasing strength of Murashige-Skoog's (MS) medium, but the length of roots was inhibited. Inter action of strength of MS medium and sucrose concentration was examined. High concentration of both components stimulated bulb lei growth, but the second strength of MS medium containing 90 or 120 g/l sucrose stimulated callus induction and inhibited the growth of bulblets. Maximum growth took 100 days for bulblets and about 50 days for roots. The change of fresh weight/dry weight ratio during differentiation is also discussed.     

Stimulating effect of spermine on bulblet formation in bulb-scale segments of Lilium longiflorum

When bulb-scale segments of Lilium longiflorum were cultured on a medium containing auxin and cytokinin, the proportion of the expiants with newly-formed bulblets was significantly increased by the application of different polyamines. The most effective polyamine was spermine, where more than 90% of segments formed an average of 5 bulblets as compared to controls where less than 50% explants formed an average of 1.5 bulblets. Application of arginine one of the precursors putrescine biosynthesis, slightly promoted bulblet formation. The putrescine-stimulated bulblet formation was strongly inhibited by simultaneous addition of an inhibitor of the spermidine synthase, cyclohexylamine. The spermidine-promoted bulblet formation, however, could not be suppressed by this inhibitor. The promotive effect of spermidine on bulblet formation was reversed by an inhibitor of the spermine synthase, N-(3-aminopropyl)cyclohexylamine, but application of this inhibitor with spermine did not show any apparent effect on the bulblet formation. Endogenous level of spermine increased in common during bulblet formation that were stimulated by exogenous polyamines. Thus, spermine seemed to be the main stimulating chemical on bulblet formation in lily bulb-scale segments.Abbreviations APCHA N-(3-aminopropyl)cyclohexylamine - Arg arginine - BA benzyladenine - CHA cyclohexylamine - MS Murashige and Skoog's - NAA naphthaleneacetic acid - Orn ornithine - Put putrescine - Spd spermidine - Spm spermine     

Lilium candidum bulblet and meristem development

Lilium candidum L., commonly known as the Madonna lily, is a wild Lilium species with medicinal properties and excellent potential as an ornamental crop, but one that has been scarcely investigated. The aim of this research was to study (1) tissue culture propagation of L. candidum bulblets, (2) early bulblet development, and (3) the effect of temperature and bulblet weight on bulblet and plant growth and meristem development. An investigation of the effect of explant type and temperature on in vitro bulblet propagation showed that scales were the most efficient explants for in vitro propagation and that exposing the regenerating bulblets to 15°C for 4 wk increased bulblet weight but reduced the number of bulblets produced. For bulblets planted in soil after 12 wk of exposure to 15°C or 25°C, the fastest growth was observed in the bulblets that had been exposed to 15°C and that had a larger initial size. Histological examination showed that young in vitro-grown bulblets had a rudimentary meristem comprising few cells with no layer organization. After 12 wk of growth, all bulblets showed a layered meristem, regardless of bulblet size or exposure to 15°C. However, an increased amount of leaf primordia was detected in larger bulblets. Furthermore, the histological examination revealed that in L. candidum, as opposed to other lily species, there had been no real "phase change" in the meristem and that the phase change from juvenile to vegetative adult occurred at a much later stage in L. candidum than in other species.     

<Emphasis Type="Italic">Narcissus</Emphasis> bulblet formation <Emphasis Type="Italic">in vitro</Emphasis>: effects of carbohydrate type and osmolarity of the culture medium

Shoot clump cultures of Narcissus cultivars St. Keverne and Hawera were used to investigate the effects of culture medium carbon supply, type of carbohydrate and osmolarity on in vitro bulblet development. Increasing the medium osmolarity using mannitol or sorbitol, which did not act as substrates for growth, failed to stimulate bulblet formation with either cultivar. An exception to this was a relatively small increase in total bulblet dry weight per culture, in the cultivar Hawera only, caused by adding 30 g l ^–1 sorbitol in combination with 30 g l^–1 sucrose. Simultaneously increasing the medium osmolarity and carbon supply using the metabolisable carbohydrate sources, sucrose, glucose, fructose or an equimolar mixture of glucose and fructose stimulated bulblet production, total dry matter accumulation and partitioning into bulblets. At comparable levels of carbon supply up to 19.0 g l^–1, bulblet development of both cultivars was similar with monosaccharide and sucrose media. This indicates that substrate supply is more important for bulblet development than osmolarity of the culture medium. The cultivar Hawera also showed similar responses to monosaccharide and sucrose media supplying 37.9 g C l^–1, despite the high osmolarity of monosaccharide media (c. 650 m Osm kg^–1, equivalent to –1.6 MPa, compared to 380 m Osm kg^–1 for sucrose medium). However in St. Keverne total dry matter accumulation and dry weight per bulblet were further stimulated only by increasing the sucrose supply from 19.0 to 37.9 g C l^–1, not by increasing the monosaccharide supply. Implications of the findings for Narcissus micropropagation are discussed.     

VEGETATIVE DISPERSAL OF ALLIUM NEAPOLITANUM

Vegetative propagation and dispersal were studied in attached and detached increase bulblets of Allium neapolitanum. The bulblets were sown in 3 positions (upright, horizontal, and inverted) at various depths, and the directions of the contractile roots and movement in the soil were calculated. A thick contractile root develops spontaneously from the base of the bulblet, irrespective of planting level, even at normal depth. The root is ageotropic and its direction is chiefly determined by the bulblet position. It generally develops at a plane perpendicular to the longitudinal axis of the bulblet. Thus, in upright and inverted bulblets, the root is horizontal, while in horizontal ones it may grow in various directions but always on the plane perpendicular to the axis of the bulblet. Depth has a marked effect on the direction of the contractile roots, diverting them from their original route (upwards in deep-seated plants and downwards in shallow ones). Thus, the vegetative dispersal of Allium neapolitanum is tridimensional, within definite soil levels.     

High frequency of shoot multiplication and bulblet formation of garlic in liquid cultures

In vitro shoot proliferation and bulblet production of garlic (Allium sativum L.) was studied in liquid cultures. Shoots grown in vitro were used as explants and were cultured in MS medium supplemented with 2% (w/v) sucrose and 0.5 mg l^–1 2-iP. Three culture methods (semi-solid, liquid-immersion and raft) were compared for shoot proliferation. Explants in liquid (immersion) culture exhibited an increased multiplication rate and fresh weight of shoots after 3 weeks of culture as compared with the other treatments. Bulblet formation and growth were studied in liquid medium with different concentrations of sucrose (2–13%). MS medium containing 11% (w/v) sucrose was optimal for bulblet development and bulblets developed in this medium within 9 weeks in culture. The highest multiplication rate was (135 bulblets/explant) found when explants were cultured in bulbing medium (MS medium containing 0.1 mg l^–1 NAA+11% (w/v) sucrose) supplemented with 10 M JA. Growth retardants CCC, B-9, ABA also promoted induction and growth of bulblets. Darkness promoted the bulblet induction and growth compared to light conditions (16-h photoperiod of 50 mol m^–2 s^–1). The dormancy of bulblets was broken by cold treatment at 4 °C for 8 weeks.     

细叶百合无性繁殖条件的选择

以栽培的2 年生细叶百合(Lilium pumilum DC.)鳞茎为扦插材料, 将鳞茎分内、中、外三层剥取其鳞片, 观察鳞片在不同温度、光照强度、基质中的扦插生小鳞茎的效果。扦插40d 时, 鳞片基本枯萎, 此时的实验结果是:①影响扦插效果的重要因子是温度和光强, 25℃高温避光生鳞茎最佳;其次是鳞片位置, 中鳞片和外鳞片好于内鳞片;基质对扦插影响不大。②每百鳞片生小鳞茎数和小鳞茎直径呈正相关。③相同条件下, 相同部位的刀切鳞茎段所产生小鳞茎数量明显高于手掰的整片鳞片, 这一现象至今未见报导。