蝴蝶兰原球茎增殖分化影响因子探讨

采用3个月胚龄的种胚播种形成的原球茎,初代培养时间在2.5~3个月之间换瓶转接,原球茎生长状况较好,分化形成的芽苗生长健壮;以1/2MS为基本培养基培养30d,原球茎增殖率达332%,MS培养基更有利于分化,培养2个月分化发芽率达90.6%;附加不同细胞分裂素原球茎的增殖率为6-BA >KT >Ad+KT >Ad;NAA浓度在0~1.0mg/L,对原球茎增殖分化影响不明显。     

安诺兰无菌播种组培快繁技术研究

安诺兰(Anota hainanensis)种子无菌播种试验表明:在MS+6-BA 0.1 mg/L培养基上种子能形成原球茎团;1/2 MS培养基有利于原球茎增殖,加6-BA 1.0 mg/L+NAA 0.1 mg/L原球茎增殖效果较佳,增殖率达793.3%;在1/2 MS+IBA 1.0 mg/L+2.0 g/L活性碳+100 g/L香蕉匀浆+100 ml/L椰子水的培养基上,试管苗生根效果最好;试管苗移栽至椰糠上,成活率达91.4%。     

兰属"小金童"组织培养研究

兰属小金童(Cymbidium Golden elf. 'Sundust')茎尖组织培养研究表明:茎尖外植体在不添加植物生长调节剂的花宝1号培养基上诱导产生原球茎;原球茎在花宝1号3g/L + 6-BA 1.5mg/L的液体培养基上培养,能形成快速增殖的丛生原球茎;不同的培养方式对原球茎的增殖效果有显著差异;原球茎在不附加植物生长调节剂的1/2MS或花宝1号培养基上分化成无根小苗,然后在花宝1号+NAA 0.2mg/L+IBA 0.6mg/L培养基上诱导生根,进而再形成完整植株.     

兰属“小金童”组织培养研究

兰属小金童(Cymbidium Golden elf.‘Sundust’)茎尖组织培养研究表明:茎尖外植体在不添加植物生长调节剂的花宝1号培养基上诱导产生原球茎;原球茎在花宝1号3g/L 6-BA 1.5mg/L的液体培养基上培养,能形成快速增殖的丛生原球茎;不同的培养方式对原球茎的增殖效果有显著差异;原球茎在不附加植物生长调节剂的1/2MS或花宝1号培养基上分化成无根小苗,然后在花宝1号 NAA 0.2mg/L IBA 0.6mg/L培养基上诱导生根,进而再形成完整植株。     

Study on technology of aseptic sowing and rapid propagation of Demdrobium officinale

以铁皮石斛的蒴果为外植体,采用种子→原球茎→完整植株→移栽的途径快速成苗进行工厂化生产,对各阶段培养基进行筛选,以及其他一些影响因子进行比较研究.结果表明:人工授粉后生长60～180 d的铁皮石斛种子在离体条件下均能萌发,其中授粉150～180 d种子的萌发效果最好,萌发率为87.2%～94.4%,适宜的萌发培养基为MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+马铃薯汁200 g/L+AC 1.0 g/L;原球茎增殖的最佳培养基为MS+6-BA 1.5 mg/L+NAA 0.1 mg/L+香蕉汁100 g/L+AC 1.0 g/L,繁殖系数约为20倍/50 d;原球茎在MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+马铃薯汁200 g/L+AC 1.0 g/L培养基上进行分化培养,分化的同时还能进行一定的增殖;将已分化的芽苗转接到壮苗培养MS+6-BA 0.5 mg/L+NAA 0.2 mg/L+香蕉汁100 g/L+AC 1.0 g/L上培养1代后,转接到生根培养基1/2MS+NAA 0.8 mg/L+无机盐A 0.2～0.5 mg/L +香蕉汁100 g/L+AC 1.0 g/L上,培养50～70 d后,生根率100%,无机盐A可以有效地控制愈伤或原球茎的形成,明显提高生根苗的数量和质量.在桂林地区,生根苗以3～5月和9～10为最佳移栽期,以通过高温处理并堆沤腐熟的松树皮为基质,移栽成活率可达90%.     

铁皮石斛无菌播种产业化繁育技术研究

以铁皮石斛的蒴果为外植体,采用种子→原球茎→完整植株→移栽的途径快速成苗进行工厂化生产,对各阶段培养基进行筛选,以及其他一些影响因子进行比较研究。结果表明:人工授粉后生长60～180d的铁皮石斛种子在离体条件下均能萌发,其中授粉150～180d种子的萌发效果最好,萌发率为87.2%～94.4%,适宜的萌发培养基为MS+6-BA 1.0mg/L+NAA 0.1mg/L+马铃薯汁200g/L+AC 1.0g/L;原球茎增殖的最佳培养基为MS+6-BA 1.5mg/L+NAA 0.1mg/L+香蕉汁100g/L+AC 1.0g/L,繁殖系数约为20倍/50d;原球茎在MS+6-BA 1.0mg/L+NAA 0.1mg/L+马铃薯汁200g/L+AC 1.0g/L培养基上进行分化培养,分化的同时还能进行一定的增殖;将已分化的芽苗转接到壮苗培养MS+6-BA 0.5mg/L+NAA 0.2mg/L+香蕉汁100g/L+AC 1.0g/L上培养1代后,转接到生根培养基1/2MS+NAA 0.8mg/L+无机盐A 0.2～0.5mg/L+香蕉汁100g/L+AC 1.0g/L上,培养50～70d后,生根率100%,无机盐A可以有效地控制愈伤或原球茎的形成,明显提高生根苗的数量和质量。在桂林地区,生根苗以3～5月和9～10为最佳移栽期,以通过高温处理并堆沤腐熟的松树皮为基质,移栽成活率可达90%。     

云南火焰兰种子无菌萌发和试管育苗

对云南火焰兰进行无菌播种技术试验.结果表明,适宜萌动的培养基为1/4MS 椰子水20% 蛋白胨1 g L-1 蔗糖10 g L-1,播种30 d时萌动率可达93.1%;适宜萌发的培养基为VW 椰子水10%,培养75 d时萌发率达36.6%.盐浓度较高的MS培养基不适合种子萌发和原球茎生长,低盐的KC培养基上种子能较好地萌发和成苗;添加椰子水可明显提高种子萌动率,并有利于原球茎的生长和分化.光培养和暗培养对种子萌动的影响差异不明显,但暗培养3周会对种子后期发育不利.添加150 g L-1的苹果匀浆有利于幼苗生长.试管苗移栽成活率在95%以上.     

同色兜兰的非共生萌发与快速繁殖研究

以授粉后不同发育时期的同色兜兰种子为材料,观察其形态特征和萌发过程,并探讨建立同色兜兰高效快繁体系的最佳条件。结果表明,种子发育中后期即授粉后210~240d为较适宜的采收期,授粉后210d的种子萌发率最高(达77.79%);1/4 MS和1/2MS为同色兜兰适宜的基本培养基,添加100mL/L椰乳或1g/L蛋白胨对种子萌发及原球茎生长和分化有明显的促进作用;添加1g/L活性炭对原球茎褐化有一定的抑制作用,但添加剂量不宜过大;添加香蕉汁和苹果汁对同色兜兰种子萌发和原球茎生长分化有抑制作用;暗处理对同色兜兰种子萌发无影响;分化后的原球茎在壮苗和生根培养基上培养120d即可得到4~5片叶、高3~5cm的同色兜兰健壮试管苗。     

白芨胚发育与种子萌发的关系

随着白芨胚龄增大,胚成熟度不断增加,有胚率和萌发率逐渐提高,种子萌发时间逐渐变短。胚龄为20周后采收最好,有胚率最高,萌发率为100%,萌发时间只需1周。种子萌发时胚先发育成原球茎,原球茎再分化出叶片和根系,形成完整的小植株;花宝一号1g/L+花宝二号2g/L+10%椰子汁培养基可促进种子萌发及幼苗生长,比常用的基本培养基加生长调节剂更简捷,幼苗生长健壮。     

西藏虎头兰高效植株再生体系的研究

为建立西藏虎头兰(Cymbidium tracyanum)的高效快速繁殖体系,该研究以野生西藏虎头兰种子为外植体,通过分析不同基本培养基和植物激素配比对原球茎诱导、增殖和分化的影响,以及光照时间和培养温度对试管苗生长的影响,筛选出适宜西藏虎头兰植株高效再生的条件。结果表明:适宜西藏虎头兰生长的基本培养基为1/2 MS;种子萌发和原球茎诱导的最适培养基为1/2 MS+1.0 mg·L~(-1)6-BA+0. 5 mg·L~(-1)NAA,培养50 d后,有95.00%的种子发育成原球茎;原球茎增殖的最适培养基为1/2 MS+2.0 mg·L~(-1)NAA,培养30 d,增殖倍数为4.25;原球茎的最适分化培养基为1/2 MS+2.0 mg·L~(-1)NAA+60 g·L~(-1)土豆泥+0.5g·L~(-1)活性炭,培养10 d,不定芽发生率为98.33%,培养40 d,幼苗生根率为94.67%;试管苗在温度20℃、光照时间12 h·d~(-1)、光照强度2 000 lx的条件下培养,苗长势好,叶片生理性焦尖发生率仅为3.33%;以腐殖土作为栽培基质,试管苗的移栽成活率为97.78%。该研究结果为保护西藏虎头兰野生资源和工厂化育苗提供了科学依据和技术支持。     

丽格海棠叶片离体培养与工厂化育苗体系的建立

利用丽格海棠叶片进行离体培养,建立工厂化育苗体系,B₅培养基适合于叶片诱导分化丛芽。丛芽在B₅+BA 0.5mg/L+IBA 0.05mg/L培养基中,增殖系数达6.06;B₅+IBA 0.5mg/L中,生根率达100%,平均根数4.5条。生根前无生长调节剂壮苗是获得健壮植株的有效方法。培养基中附加低浓度蔗糖对维持植株正常形态起关键作用。     

蓝猪儿组织培养和再生

光照有利于蓝猪儿(Torenia fournieri)种子的萌发。上胚轴转接在生芽培养基(MS+0.1 mg/L 4PU+2 mg/L NAA)上,25℃,光暗16/8h培养18d,芽发生,25d不定芽数目达7~8个。叶或茎形成愈伤组织的适宜培养基为MS+0.05 mg/L 2,4-D+0.2 mg/L NAA,愈伤组织再分化形成芽的能力较弱。1/2 MS培养基无论是否添加萘乙酸,蓝猪儿试管苗都能发生不定根,萘乙酸使不定根的数目增多,分布密集。     

杂交兰类原球茎增殖中芽分化的控制和快速繁殖

以‘玉女杂交兰’为材料,研究了培养时间、活性炭、切块大小和外源生长调节物质对类原球茎增殖和分化的影响。结果表明,培养时间、活性炭和切割处理对类原球茎增殖和分化影响显著,在培养基中添加活性炭或对类原球茎进行切割均能有效控制增殖过程中的芽分化。将类原球茎切成直径2~3 mm的小块接种到培养基MS＋1.0 mg.L-16-BA＋0.2 mg.L-1NAA＋0.3 g.L-1AC＋30 g.L-1蔗糖＋5.5 g.L-1琼脂中培养40 d,类原球茎增殖率为384.23%。将增殖后的类原球茎接种到培养基1/2MS＋1.5 mg.L-16-BA＋0.1 mg.L-1NAA＋20 g.L-1蔗糖＋5.5 g.L-1琼脂中培养40 d,芽分化率为463.06%。将分化的芽转入培养基1/2MS＋0.5 mg.L-1NAA＋0.5 g.L-1AC＋20 g.L-1蔗糖＋100 g.L-1土豆汁＋5.5 g.L-1琼脂中培养,生根率为100%,平均根分化数为2.80条.株-1。以泥炭土为基质,组培苗的移栽成活率可达97.78%。     

Regeneration of plantlets from in vitro raised leaf explants of Cleisostoma racimeferum Lindl

Protocorm like bodies (PLBs), callus and shoot buds developed in culture from in vitro raised foliar explants of Cleisostoma racimeferum. Among the different basal media, better result was obtained on MS medium containing sucrose (3%) and BA (2 microM) with approximately 80% frequency after 40 days of culture. Young leaves (15 week old) produced better PLBs. Whole leaf placed vertically upside-up orientation can regenerate PLBs and shoot buds (80%). PLBs and shoot buds formed on entire surface of the leaves. Cultures on BA and NAA (2 and 2 microM respectively in combination) stimulated callus mediated regeneration (68%). The rooted plantlets regenerated within 8-10 week from PLBs and shoot buds on MS medium containing IAA and kinetin (2 microM each in combination). BA containing medium triggered multiple shoot bud formation, while NAA alone or in combination with other growth regulators was inhibitory. Incorporation of activated charcoal (0.01%) in the medium stimulated formation of repetitive PLBs and multiple shoot buds. Rooted plants were ready for harvest after 20-22 week of initiation of culture. About 65% of the potted plants survived after 3 months in the poly house.     

正交实验法在香果树种质离体保存中的应用

以香果树带芽茎段为外植体,采用正交实验法研究活性炭、琼脂、封口材料和培养容器对香果树试管苗离体保存的影响。结果表明,香果树带芽茎段用200 ml三角瓶,无菌培养容器封口膜封口,保存于MS+KT 2.0 mg/L+6-BA 2.0 mg/L+NAA 0.1 mg/L+30 g/L蔗糖+0.5 g/L活性炭+7.5 g/L琼脂培养基上6个月后,成活率可达86.3%。离体保存的试管苗没有发生遗传变异。     

怀山药类原球茎的诱导形成与植株再生

为提高山药离体繁殖的速度, 缩短繁殖周期, 以铁棍山药(Dioscorea opposita cv. ‘Tiegun’)带腋芽茎段为材料, 对类原球茎的诱导、增殖、分化与植株再生进行了研究。结果表明, 铁棍山药类原球茎诱导的最适培养基为MS+1.0 mg·L^-1 TDZ+30 g·L^-1蔗糖, 增殖的最适培养基为MS+9 mg·L^-1 6-BA+30 g·L^-1蔗糖, 分化的最适培养基为MS+2 mg·L^-1 KT+0.02 mg·L^-1 NAA+30 g·L^-1蔗糖, 最适生根培养基为1/4MS+0.05 mg·L^-1 NAA+1.0 mg·L^-1 PP333+15 g·L^-1蔗糖, 生根率达80%, 移栽成活率可达85%。类原球茎的诱导形成及植株再生体系的建立为怀山药种苗的快速繁殖提供了一条新途径。     

Plant regeneration via organogenesis from adventitious bud explants of a medicinal herb species, <Emphasis Type="Italic">Polygonatum cyrtonema</Emphasis>

Summary Explants derived from adventitious buds, rhizomes, stems, and leaves of a medicinal plant, Polygonatum cyrtonema, were studied for plantlet regeneration, and only adventitious bud explants were able to be regenerated into plantlets. Regeneration was also accompanied by the formation of rhizome-like tissue, the medicinal portion of the plant. The optimum hormone combination for plantlet regenertion was 4.44 μM benzyladenine plus 2.26 μM 2,4-dichlorophenoxyacetic acid, at which new adventitious buds were obtained from 96.6% of the adventitious bud explants, with an average of 5.2 buds per explant. The best medium for root induction was half-strength Murashige and Skoog medium with 4.57 μM α-naphthaleneacetic acid, as 92% of regenerated buds rooted. Regenerated plantlets were successfully transferred to a greenhouse with 86% survival. Histological observation indicated that new adventitious buds originated from the superficial meristematic cell cluster of the granular callus induced from adventitious bud explants via organogenesis.     

江西铅山红芽芋脱毒苗球茎愈伤组织诱导及其再生体系的建立

以江西铅山红芽芋脱毒苗为试材,研究不同因素对红芽芋脱毒苗球茎愈伤组织诱导及其再生体系的影响,以期对红芽芋脱毒苗的再生体系进行优化。结果表明,红芽芋脱毒苗球茎愈伤组织诱导的最佳培养基是MS+TDZ 2 mg·L^-1+2,4-D 1 mg·L^-1。红芽芋脱毒苗球茎愈伤组织分化的最佳培养基是MS+TDZ 2 mg·L^-1+NAA 1 mg·L^-1。红芽芋脱毒苗不定芽生根的最佳培养基是1/2MS+NAA 0.5 mg·L^-1+PP₃₃₃ 0.5 mg·L^-1。红芽芋再生苗最好的移栽基质为发酵后的腐锯木屑。红芽芋脱毒苗球茎愈伤组织再生苗移栽时最佳的PP₃₃₃浓度为20～50 mg·L^-1。本试验成功建立了红芽芋脱毒苗球茎愈伤组织的再生体系,为红芽芋脱毒苗转基因的研究和种质创新奠定了基础。     

Mass multiplication of protocorm-like bodies using bioreactor system and subsequent plant regeneration in Phalaenopsis

Protocorm-like bodies (PLBs) formed on leaf segmentsin vitro were used as explants for bioreactor cultures. Continuous immersion cultures (air lift column and air lift-balloon bioreactor), and temporary immersion cultures (with or without charcoal filter attached) were used for the culture of PLB sections. A temporary immersion culture with charcoal filter attached was most suitable for PLB culture. About 18,000 PLBs were harvested from 20 g of inoculum (∼1000 PLB sections) in 2 l Hyponex medium after 8 weeks of incubation. Aeration in a bioreactor at 0.5 or 2.0 volume of air per volume of medium min⁻¹ (vvm) yielded similar levels of biomass production. PLBs grown in bioreactors were cultured on solid Murashige and Skoog, Vacin and Went, Knudson C, Lindemann and Hyponex media. Hyponex medium was found to be suitable for conversion of PLBs into plantlets and 83% of PLBs transformed into plantlets on this medium. The feasibility of using PLBs for large-scale micropropagation was evaluated for scaled-up liquid cultures in bioreactors, rate of proliferation, and regeneration. This revised version was published online in June 2006 with corrections to the Cover Date.     

牛蒡组织培养和高频植株再生的研究

通过对牛蒡(A rctium lapp a L.)不同外植体、不同激素配比的比较研究,建立了牛蒡离体培养高效植株再生体系.牛蒡子叶与下胚轴切段在含2.0 m g/L 2,4-D和0.5~2.0 m g/L BA的M S培养基中愈伤组织诱导率可以达到87%~100%;在1.0~3.0 m g/L NAA和0.5~2.0 m g/L BA的M S培养基上通过愈伤组织间接分化或外植体直接分化形成不定芽,其中愈伤组织分化率可达100%;下胚轴的分化率明显高于子叶,在1.0 m g/L NAA和1.0 m g/L BA的M S培养基上下胚轴直接分化率达77.3%.组织学观察发现牛蒡再生有器官发生和体细胞胚发生两种途径.将生长状态良好的不定芽转至含1.0 m g/L IBA和1.0 m g/L NAA的1/2 M S培养基上生根,移栽,成活率达到93.3%.从诱导愈伤组织到组培苗在珍珠岩中过渡成活,大约需要13周.组培苗次年开花并结实,生长形态特征正常.