新疆天山雪莲体胚诱导与分化研究

以新疆天山雪莲的叶片为外植体,分别用不同配方培养基诱导愈伤组织,后进行体胚诱导和分化培养形成再生雪莲植株.结果表明,诱导愈伤组织的最适培养基为MS 2,4-D 0.5 mg/L BA 1.5 mg/L,诱导率可达到100%;愈伤组织转移至MS 2,4-D 0.5 mg/L BA 1.5 mg/L培养基进行继代培养,增殖后的愈伤组织转移到MS 2,4-D 0.2 mg/L的液体培养基后成功诱导出雪莲体胚,出胚率达40%;将体胚接至MS ABA 0.5 mg/L培养基后,结果分化生长出大量的再生雪莲幼苗.     

从稗草花序培养诱导的体细胞胚胎发生和植株再生

在含有各种浓度2,4-D的MS培养基上,从稗草(Echinochloa crusgalli L.)的幼嫩花序切断诱导出一种白色密实,有结构的胚性组织。组织学研究和扫描电镜观察证明此白色愈伤组织系由发育阶段不同的胚状体所组成。典型的胚状体具有二极性,并带有盾片、胚芽鞘和胚根鞘等结构。此胚性愈伤组织在四个月中经过多次继代培养仍然保持其胚性性质。降低2,4-D浓度或从培养基中撤除2,4-D,可诱导大量具有根、茎、叶的完整植株。     

狗牙根颖果胚性愈伤组织的诱导和胚性细胞的超微结构及植株再生

以普通狗牙根[Cynodon dacylon(L.)Pers.cv.'Suncitv']颖果为外植体,以MS为基本培养基,外加浓度在2.0～6.0mg/L的2,4-D,能高频率地诱导出高质量的胚性愈伤组织,其中以4.0 mg/L为最佳.胚性愈伤组织最佳继代及分化的培养方法为:用MS 2,4-D 4.0mg/L继代1～2次,然后转入1/2 MS 2,4-D 2.0 mg/L中继代1～2次,再在无激素的1/2MS中光照培养10 d,最后在MS 6-BA 3.0 mg/L中诱导分化,分化成苗率达31.7%.经电镜观察发现,胚性愈伤组织结构紧密,细胞较小,内容物丰富,而非胚性愈伤组织结构疏松,细胞巨大,内含一大液泡,几无细胞器.     

影响籼稻体细胞胚胎发生几个因素的研究

以 IR36、IR50、IR52及 IR54等品种的幼穗及成熟种子为材料,研究了蔗糖浓度、2,4-D、NAA、激动素及脱落酸对体细胞胚胎发生、结构的保持及植株分化的影响。6%蔗糖有利于胚性愈伤组织的诱导;3%的有利于胚性结构的保持及植株分化。当培养基中不含2,4-D,而含激动素与 NAA 时,幼穗直接出芽;当不含激动素而含2,4-D与 NAA 时,外植体产生非胚性愈伤组织;当不含 NAA 而含2,4-D 与激动素时,外植体产生胚性愈伤组织。认为,2,4-D与激动素是籼稻体细胞胚胎发生的基本因素,而 NAA 的作用是不明显的。不同外植体(幼穗与成熟种子)的体细胞胚胎发生,对2,4-D 与激动素的反应略有不同,幼穗更为敏感。在继代培养基中,加入低浓度的脱落酸有利于胚性结构的保持。随着继代世代的延续,分化培养中愈伤组织所表现出的绿色生长点状物不能发育成完整植株。     

糜子离体体细胞胚胎发生和植株再生 Ⅱ.各种因素的影响

本试验就各种因素对糜子胚性和非胚性愈伤组织诱导的影响及其植株再生进行了较为详细的研究。结果表明,2,4-D是诱导胚性愈伤组织所必须的;蒸糖浓度、酵母浸出汁和水解乳蛋白的含量、基本培养基组成、外植体来源和黑暗培养等因素也都有不同程度的影响;而且各种因素对胚性愈伤组织的影响比非胚性愈伤组织更大。诱导胚性愈伤组织最适宜的培养基组成是MS+2,4-D(2mg/l)+BA(0.5mg/l)+蔗糖(3%)+YE(0.3%)+LH(1600mg/l)+盐酸硫胺素(0.4mg/l)。两种愈伤组织转移到无或含少量2,4-D的MS培养基上,只能从胚性愈伤组织再生植株。再生植株经移栽生长成熟并结了种子。     

亚麻品种'双亚5号'的胚性愈伤组织诱导和体细胞胚胎发生

亚麻品种'双亚5号'的种子在MS 1.0 mg·L-1 2,4-D 30 g·L-1蔗糖培养基上可诱导出愈伤组织,将其转入MS 0.5mg·L-1KT 0.5mg·L-1NAA培养基上培养10周后诱导出大量胚性愈伤组织,结构较致密,浅黄色,表面有成团的紧密粘附在一起的小颗粒形态.但其继代周期不宜太长,继代次数不宜多,否则易回到非胚性化状态.胚性愈伤组织转A.MS 1.5 mg·L-1(T 1.0 mg·L-1 2,4-D分化培养基上培养40 d后可获得大量的球形胚状体.少量球形体胚可萌发形成正常的子叶胚初期形态,较多的球形体胚形成次生体胚或仅有单极性的畸形胚状体.组织解剖学观察表明,诱导出的是亚麻胚性愈伤组织和胚状体.     

小麦遗传转化受体系统建立的研究

选用‘小偃22’和‘宁春16’小麦品种的成熟胚和幼胚进行培养,研究不同种类的胚和培养因子对愈伤组织诱导和分化的影响。结果表明,幼胚和成熟胚的愈伤组织诱导率无明显差异,但较高浓度的2,4-D有利于成熟胚的诱导,而幼胚培养时2,4-D浓度的影响效果因品种而异;两种外植体分化率的高低与KT/IAA的配比均有密切关系,但高浓度的激素水平不利于成熟胚的分化;诱导培养基中低浓度的2,4-D有利于所诱导的愈伤组织的分化。同时,在诱导培养基中添加低浓度的KT能显著提高两品种成熟胚愈伤组织的分化率;各种培养基处理与品种间都存在显著的互作效应,‘小偃22’成熟胚培养的最佳培养基组合为MSD 3.0 mg/L 2,4-D和MSD 0.5 mg/LIAA 1.0 mg/L KT,幼胚培养为MSD 4.0 mg/L 2,4-D和MSD 0.5 mg/L IAA 1.0 mg/L KT;‘宁春16’成熟胚培养为MSD 4.0 mg/L 2,4-D和MSD 1.0 mg/L IAA 1.0 mg/L KT,幼胚培养时为MSD 1.0 mg/L 2,4-D和MSD 2.0 mg/L IAA 2.0 mg/L KT。     

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。     

两个苏丹草品系成熟种子的组织培养和植株再生研究

该研究以苏丹草品系S722和Sa的成熟种子为外植体、MS培养基为基础培养基,2,4-D和NAA各3个浓度共6个处理对这两个苏丹草品系成熟种子进行愈伤诱导,探讨不同品系在不同植物生长物质浓度及植物生长物质组合中诱导愈伤组织和继代培养以及分化的能力。结果表明：苏丹草S722和Sa成熟种子的愈伤诱导率差异不显著,平均诱导率为17.19％。诱导培养基中2,4-D浓度为0.5或1 mg?L-1时,诱导效果最佳,而添加NAA不能提高愈伤诱导率。在继代培养中,设定2,4-D和6-BA各两个浓度共4个处理组合,处理1(2,4-D 1 mg?L-1＋6-BA 0 mg?L-1)的继代培养效果最佳。为了解不同植物生长物质对愈伤分化的影响,设定6-BA、NAA 各两个不同浓度、KT 3个不同浓度共5个处理组合对继代培养的愈伤进行分化培养。在5个处理中,处理1(6-BA 2 mg?L-1＋NAA 0 mg?L-1＋KT 0 mg?L-1)对 S722成熟种子诱导的愈伤分化率最高,达33.3％。在这两个苏丹草品系中,S722更容易分化培养。综上结果表明,2,4-D浓度为1 mg?L-1时诱导愈伤和继代培养效果较好,6-BA浓度为2 mg?L-1时分化效果较好。另外,针对不同苏丹草品系进行组织培养和植株再生时,适当调整植物生长物质浓度能提高植株再生的成功率。     

刺葡萄幼胚愈伤组织诱导及其高产原花青素细胞系筛选

以刺葡萄幼胚为材料,研究不同培养方式、培养基配方和培养条件对其愈伤组织诱导的影响,采用正交试验设计法筛选刺葡萄愈伤组织继代增殖的培养基配方,并对继代保持的培养条件和方式进行优化,同时进行了高产原花青素刺葡萄愈伤组织细胞系的筛选研究。结果表明,刺葡萄幼胚以平放的方式接种到MS＋1.0 mg·L^-1 2,4-D或MS＋1.0 mg·L^-1 2,4-D＋0.5mg·L^-1 KT的固体培养基上,在黑暗的条件下,能有效的诱导出愈伤组织,诱导效率为80%;刺葡萄愈伤组织继代增殖以MS＋1.5 mg·L^-1 2,4-D或MS＋1.5 mg·L^-1 2,4-D＋0.5 mg·L^-1 KT的固体培养基为佳,并且采用此两种培养基交替继代培养,在光照条件下能长期保持旺盛且生长一致的刺葡萄愈伤组织;筛选出了紫红色松脆状的高产原花青素的刺葡萄愈伤组织细胞系,培养35 d后每克鲜样的原花青素含量可达1 671.16μg。     

Embryogenic Callus Formation and Plantlet Regeneration of Piragmites communis

The present paper reports the embryogenic callus formation and plantlet regeneration of Phragmites communis. The results have been obtained as follows: The efficiency of callus induction was much higher, if reed seeds were used as explants. No dedifferentiation was observed by using leaf sheath and leaf blade as explants. The optim, um concentration of sucrose was 4% in medium. VB group and inositol had beneficial effects on callus growth. But yeast extract inhibited callus induction and callus growth markedly. For this inhibited reaction, the higher concentration, the more obviously the callus growth was inhibited. Higher levels of 2,4-D had unfavourable effects on callus growth in callus subculture. The concentration of 2,4-D in dedifferentiation medium had relation to embryogenic callus formation. Embryogenic callus had higher frequency of differentiation for long-term subculture. On the other hand, nonembryogenic callus most often lost their morphogenetic competence. Authors found that the surface structure of the two types of calluses was different by means of observation by scanning electron microscope. The peroxidase and the esterase isoenzyme pat- terns, as well as the soluble protein of both types of calluses were different too.     

百合体细胞胚胎发生和植株再生

以切花百合(Lilium)品种‘黄天霸’(‘Manissa’)花器官为外植体诱导体细胞胚胎发生与植株再生。结果表明,不同花器官、不同激素配比对愈伤组织形成均具有显著影响。花丝为最佳外植体,激素对愈伤组织诱导的影响效应为NAA>6-BA>2,4-D,最适培养基为MS+1.0 mg.L-1NAA+0.2 mg.L-16-BA;激素诱导体细胞胚胎发生的影响效应为2,4-D>KT>6-BA,最佳培养基配方为MS+1.0 mg.L-12,4-D+0.2 mg.L-1KT+1.0 mg.L-16-BA;MS培养基添加IBA可促进体细胞胚萌发成苗,体细胞胚芽成苗的最佳培养基为MS+0.2 mg.L-16-BA+1.0 mg.L-1IBA。     

苦丁茶愈伤组织的诱导与褐变抑制

以叶片为材料,对苦丁茶愈伤组织的诱导,继代培养及褐变调控的研究表明：（１）苦丁茶愈伤组织的诱导及继代培养均以ＭＳ附加ＢＡ ２．０ｍｇＬ＾－１和ＮＡＡ ４．０ｍｇＬ＾－１的培养基效果最好;（２）０．１％的植酸可明显促进愈伤组织生长、抑制褐变,而硫代硫酸钠效果最差;（３）连续培养４０－５０ｄ愈伤组织增长倍数达到最大值;（４）继代２７次以后愈伤组织生长速度开始下降。这些条件为下一步细胞培养生产苦丁茶甙等     

长柄双花木愈伤组织诱导及植株再生

以长柄双花木当年生嫩梢上的叶柄、嫩茎、嫩叶为外植体,对影响长柄双花木愈伤组织诱导和继代、分化主要因素进行研究。结果表明:在培养基MS+NAA0.5mg/L+2,4-D2.0mg/L上,三种外植体均可诱导出愈伤组织,其中叶片愈伤组织诱导率最高。该培养基还可作为愈伤组织继代培养基,但继代培养周期不超过2周。愈伤组织接种在MS+BA2mg/L上分化不定芽,根的诱导在1/2MS+IBA0.5mg/L培养基上进行。     

Plant regeneration from callus cultures in two ecotypes of reed (Phragmites communis Trinius)

Summary Different ecotypes of reed (Phragmites communis Trinius) provide an ideal resource for studies on plant environmental adaptations and presence of genes relating to stress resistance. Dune reed is a drought-tolerant reed ecotype growing in the desert regions of north-west China. In this work, in vitro culture systems of dune reed and local swamp reed (as control) were established by optimizing the culture conditions for each of them. Bright yellow calluses were induced on a Murashige and Skoog medium containing 4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 5.4 μM naphthaleneacetic acid and 2.2μM benzyladenine. Benzyladenine promoted callus induction, but was not required for callus maintenance. Four types of callus have been identified from each of the reed ecotypes. Two types of callus, i.e. type A (formed normal green shoots) and type C (formed albino plants), were both found as embryogenic calluses. The optimal concentrations of 2,4-D to maintain embryogenic callus were 2.3–4.5 μM for dune reed and 9.0–13.5 μM for swap reed. Plant regeneration was achieved from types A and C callus in a hormone-free medium. The embryogenic calluses of swamp reed have been maintained for over 2 yr and still retain their strong embryogenic potential; however, those of dune reed gradually lost their embryogenic potential after only 7 mo. of culture. Regenerated plants from the two reed ecotypes showed, after a growth season, similar morphology and the same chromosome number (2n=8x=96, octoploid) as the wild plants.     

Influence of form of activated charcoal on embryogenic callus formation in coconut (<Emphasis Type="Italic">Cocos nucifera</Emphasis>)

Development of micropropagation protocols for Cocos nucifera has progressed slowly. Activated charcoal is included in the culture medium of each protocol, mainly to prevent tissue browning. Charcoal production procedures can affect the properties of different brands. In this study, eight types of activated charcoal were evaluated for their effects on free 2,4-dichlorophenoxyacetic acid level, pH, conductivity, and osmolarity of the culture medium and on the frequency of embryogenic callus induction. Moreover, the effect of particle size of the optimum charcoal type on embryogenic callus development was also studied. Charcoal type had a significant effect on (Y3) culture medium properties. Free 2,4-D was highest in Reactivos y Productos Químicos Finos-containing medium and pH was lowest in MERCK-containing medium. Charcoal type also influenced embryogenic callus induction, with acid washed for plant cell and tissue culture-, DARCO- and United States Pharmacopeia-containing media promoting ~60% embryogenic callus, but with different optimal 2,4-D concentrations. Particle size profiles varied among all charcoal types, although small particle fraction (<38 μm) was abundant in all. Use of small particle fractions produced higher frequencies of embryogenic callus (70%) than either large particle or whole charcoal fractions.     

Somatic embryogenesis and plant regeneration from cell suspension and tissue cultures of mature himalayan poplar (Populus ciliata)

Somatic embryogenesis and plantlet formation were obtained from callus and cell suspension cultures of 40-year- old Himalayan Poplar (Populus ciliata Wall ex Royle). Callus and cell suspensions were obtained by transfer of inoculum of semiorganized leaf cultures, which were maintained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP), to MS with 2,4-dichlorophenoxyacetic acid (2,4-D). Reduction of 2,4-D concentration during subsequent subculture of cell suspensions resulted in the formation of embryoids. These embryoids developed further only after being transferred to agar-based MS medium supplemented with BAP and naphthalene acetic acid. Loss of embryogenic potential was observed in cell suspensions after 6 subcultures. However, callus cultures retained the embryogenic potential even after repeated subcultures for more than a year. Plantlets could be successfully hardened and grown in natural outdoor conditions.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthalene acetic acid - MS Murashige and Skoog (1962) medium     

The role of auxins in differentiation of rice tissues culturein Vitro

Effects of various auxins on callus induction (dedifferentiation) and organ redifferentiation from the callus were studied by using various tissues of rice,Oryza sativa L. cv. Kyoto Asahi. 2,4-D, NAA and IAA were used as auxins for the test of their ability to induce callus. All of these were active. This callus induction by auxin was successful in all tissues used; seed, root, shoot nodule, anther and ovary. In all of the calluses induced by various auxins such as 2,4-D, NAA and IAA and derived from various tissues such as seed, root, shoot nodule, anther and ovary, organ redifferentiation, i.e., formation of shoots and roots was achieved by removing the auxins from the medium used for the callus calture. Cytokinins were not necessary for the organ redifferentiation in these calluses. These results suggest that auxin is the only exogenous factor that determines dedifferentiation and redifferentiation in rice plant tissues culturedin vitro.     

植物生长调节剂对南方红豆杉愈伤组织培养和紫杉醇合成的影响

通过不同种类和水平植物生长调节剂对南方红豆杉(Taxus chinensisvar.mairei)愈伤组织诱导、生长和紫杉醇合成能力影响的研究发现:诱导培养初期,以无植物生长调节剂的MS为基本培养基,在附加不同植物生长调节剂组合作用下愈伤组织产生的时间和生长、在相同植物生长调节剂组合作用下不同外植体愈伤组织的产生时间和生长均表现出较显著差异,2,4-D/NAA高于0.4时,不利于南方红豆杉愈伤组织的诱导。转换到附加不同植物生长调节剂组合的B5培养基上后,随培养继代次数的增加,生长差异逐渐缩小,直至不显著,表明参考不同文献报道最优配方所设计的各植物生长调节剂组合对南方红豆杉愈伤组织的生长均较适宜,有利南方红豆杉愈伤组织生长的植物生长调节剂优化组合没有唯一性。但不同调节剂组合作用下的同源愈伤组织中、相同调节剂作用下不同源愈伤组织中紫杉醇含量均存在着极显著差异,适当水平(2 mg/L)的2,4-D单用,或与适当水平的KT、6-BA、KT GA配合使用,对南方红豆杉愈伤组织紫杉醇的合成较有利,NAA则不太有利,幼茎和叶愈伤组织产紫杉醇的水平较其它愈伤组织为高。