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1.
地生兰(Cymbidium)个体发生途径研究   总被引:1,自引:0,他引:1  
地生兰的类原球茎体(PLB)在不加激素的1/2 MS培养基上驯化培养,获得“无需外源激素仍能增殖的无性系(PLB-O)”,已继代5年,不分化芽和根。连续光照有利于PLB-O分生区的增殖,8h光照的次之,黑暗中增殖速度最低。PLB培养在含10%椰乳的1/2 BMS液体培养基中,随继代培养进程,形成分枝更多的丛生型PLB,约4个月后,在PLB顶端开始形成植株。兰花离体培养中,在同一个培养物中,同时存在着处于不同发育时期的分生区、原球茎、芽和小植株。PLB培养在含1/2BMS+6BA 2mg/L+NAA 0.2 mg/L的培养基中,同一个丛生型PLB上,因分生区和PLB的发育年龄的不同,形成不同发育年龄的幼芽。  相似文献   

2.
对卡德丽亚兰种子非共生萌发条件及萌发过程中原球茎发育进行了研究。结果表明,低离子浓度培养基(KC、RE)利于卡德丽亚兰种子萌发;适宜种类和浓度的激素(1.0mg/LBA和0.1mg/LNAA或1.0mg/LKT和0.1mg/LNAA)对种子萌发有良好的促进作用。细胞学研究表明:卡德丽亚兰种子萌发过程的本质是未分化胚细胞团通过细胞的不断分裂形成原球茎,原球茎出现极性,随之分化形成芽端分生区,最终形成实生苗。  相似文献   

3.
铁皮石斛原球茎常温保存研究   总被引:2,自引:0,他引:2  
以铁皮石斛原球茎为材料,通过不同培养基、蔗糖浓度、继代周期、保存时间等多种因素对原球茎在保存过程中增殖生长和分化成苗的影响,进行铁皮石斛原球茎常温保存的研究。结果表明:铁皮石斛原球茎常温((25±2)℃)保存的适宜培养基为1/2MS、蔗糖浓度为1%,继代周期可达10个月;原球茎在5年内能保持分化和增值能力,随着保存年限的增加,分化率越来越低,可通过复壮和成苗培养提高分化成苗率。  相似文献   

4.
墨兰原球茎生长的研究   总被引:22,自引:0,他引:22  
以墨兰种子胚经培养诱导萌发形成的原球茎为材料,研究其生长的最适条件。结果表明:最佳基本培养基是MS培养基;NAA0.5mgL+BA1.0mgL-1有利于生长和分化;在培养基中加入1%蔗糖和0.1%活性炭最有利于原球茎生长;pH5.0偏酸环境适合于生长;每天给予15.5umolm-2S-116h的光照最佳。如将上述单因子组合在一起,其原球茎鲜重增长率比对照(MS培养基)高46%。在不同切割方式中,掰开法鲜重增长率比纵切法或横切法都高。  相似文献   

5.
以文心兰切花品种‘南茜’原球茎(PLB)及试管苗为材料,在培养基MS+6-BA 2 mg/L+Ad 1 mg/L+NAA 0.1 mg/L+蔗糖30 g/L基础上,利用增殖系数及不同形态指标加权分析研究马铃薯、芋头、南瓜、香蕉及活性炭(AC)5种培养基添加物及其组合对‘南茜’PLB增殖及试管苗生长的影响。结果表明,适宜文心兰PLB增殖培养的添加物为马铃薯泥50 g/L,培养45 d后,PLB增殖系数是对照的3.64倍;适宜无菌小苗(约1.5 cm)增殖及生长的添加物为香蕉泥50 g/L+AC 1 g/L,加权总分是对照的1.53 倍;适宜无菌中苗(约2.5 cm)增殖及生长的添加物为马铃薯泥25 g/L+香蕉泥25 g/L,加权总分是对照的1.55倍。  相似文献   

6.
以大花蕙兰类原球茎发生发育不同阶段的培养物为试材,采用石蜡切片法对其进行组织学研究。结果表明,类原球茎形成过程中,表层薄壁细胞首先经脱分化恢复分生能力,形成愈伤组织,随后在其表面形成瘤状突起--类原球茎;类原球茎继续发育,在其周围形成多个不规则突起,并进一步发育形成新的类原球茎;而原来的类原球茎开始进入器官分化阶段,逐渐形成完整的小植株。根据观察,类原球茎起源可能存在两条途径:体细胞胚胎发生和器官发生。  相似文献   

7.
1植物名称千代兰(Ascocenda‘Fuchs Gold'). 2材料类别幼嫩花梗. 3培养条件(1)原球茎诱导培养基:VW 6-BA 2.0mg·L-1(单位下同) KT 0.5;(2)原球茎增殖和芽分化培养基:VW 6-BA 1.0 椰乳(CM)50 mL·L-1;(3)生根培养基:1/2MS IBA 1.0 NAA 0.1 香蕉100g·L-1.上述培养基均加入2%蔗糖、0.5%琼脂,pH 5.5.培养温度(25±2)℃,光照12 h·d-1,光照度2 000 lx左右.  相似文献   

8.
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%。  相似文献   

9.
蝴蝶兰原球茎增殖分化影响因子探讨   总被引:14,自引:1,他引:13  
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。  相似文献   

10.
1植物名称小白及(Bletilla formosana). 2材料类别 种子. 3培养条件基本培养基为MS.(1)种子萌发培养基:MS KT 1.0~3.0mg.L-1(单位下同) NAA 1.0~3.0;(2)原球茎增殖培养基:1/2MS 6-BA 2.0;(3)原球茎分化培养基:1/2MS 6-BA 2.0 NAA 0.5;(4)生根培养基:1/2MS NAA 0.5.上述培养基均附加30g·L-1蔗糖、8 g·L-1琼脂,pH 5.8左右,培养温度为25℃左右,光照12 h·d-1,光照度2 0001x.  相似文献   

11.
Rapid propagation of I. malabarica (Reichb. f.) J D Hook, an endemic and endangered orchid of the Western Ghats of Kerala, India through conversion of axillary buds to protocorm-like bodies (PLBs), and subsequent plant regeneration was achieved. Growth regulators and sugar displayed significant influence in the induction of PLBs. In vitro derived shoots from field grown rhizomes of Ipsea cultured on Murashige and Skoog (MS) medium with 13.3 microM N6-benzyladenine (BA) containing 2% commercial grade sugar turned the axillary buds to PLBs within 25 days, and developed a mean of 33.1 PLBs within 50 days. Kinetin (KIN) did not induce PLBs, but facilitated axillary bud proliferation. Transfer of PLBs on medium having same concentration of BA and sugar facilitated rapid multiplication, and developed a mean of 47.5 PLBs. No decline of PLB proliferation was observed up to 10th subculture. Half strength MS medium with 6.97 microM KIN facilitated conversion of 98% PLBs to plantlets. On this media, a mean of 5.8 roots were also developed per shoot. Shoots developed bulbs during culture were grown to rhizomes. Increase of sugar to 6 or 8% hastened the development of bulbs/rhizomes. Reintroduction of PLB-derived plantlets in the natural habitat i.e. at Vellarimala (at 1300 m height) of the Western Ghats of Kerala was attempted as a means to assist in situ conservation. This is the first report of conversion of axillary buds to PLBs. The protocol enables to surmount the threat of extinction of this endemic and endangered orchid.  相似文献   

12.
A mixed culture comprised of both embryonic globules and nonembryogenic callus. was derived from seedling hypocotyls of Daucus carota cv. Scarlet Nantes on 2,4-D-containing medium using well-established methods. Then the mixed cultures were transferred to, and serially subcultured on, a hormone-free medium near pH 4. The medium contained 1 m M NH+ as the sole nitrogen source. When cultured in this way, embryonic globules were able to multiply without development into later embryo stages Nonembryogenic callus did not survive. Continuous culture of embryonic globules on this low pH hormone-free medium yielded cultures consisting entirely of preglobular stage proembryos (PGSPs). PGSP cultures have been maintained as such with continuous multiplication for nearly 2 years without loss of embryogenic potential. These hormone-free-maintained PGSPs continue their development to later embryo stages when cultured on the same hormone-free medium buffered at pH 5.8. We show that hormone-free medium near pH 4 can replace 2.4-D in its ability to sustain multiplication of 2,4-D-initiated embryogenic cells of carrot at an acceptable growth rate without their development into later embryo stages. This procedure provides selective conditions that do not permit the growth of nonembryogenic cells while providing an adequate environment for embryogenic cell proliferation and should prove invaluable in studying habituation.  相似文献   

13.
Large-scale in vitro propagation protocol for Dendrobium hybrids Sonia 17 and 28, two highly prized commercial cut flower cultivars through shoot multiplication using flower stalk node explants and protocorm-like bodies (PLBs) formation was accomplished. Both hybrids did not exhibit significant differences in initiation, multiplication, rooting, and field establishment. Flower stalk nodes cultured on half strength Murashige and Skoog (MS) medium supplemented with 6.97 microM kinetin (Kn), or 15% coconut water (CW) or 13.3 microM of N6-benzyladenine (BA) evoked bud break. Kn showed better growth of the initiated bud. Excision and culture of the initiated shoots on medium having same amount of Kn developed more than 5 shoots per shoot directly from the base. Subsequent culture enhanced the rate of shoot induction. Transfer of isolated shoots onto 44.4 microM of BA enriched medium displayed induction of more than 6 PLBs from the base within 60 days. PLBs underwent rapid multiplication upon transferral to medium having the same concentration of BA (44.4 microM). Subsequent culture increased the proliferation of PLBs. No decline was observed in the proliferation of shoots as well as PLBs up to 15th subculture. PLBs transferred onto half strength MS medium with 6.97 microM of Kn underwent conversion of more than 90% PLBs to shoots. The shoots were rooted at the best on half strength MS medium with 2 g l(-1) activated charcoal. Survival rate of the plantlets of the two hybrid cultivars after acclimatization was more than 80%.  相似文献   

14.
Genetic transformation of Cymbidium orchid by particle bombardment   总被引:13,自引:0,他引:13  
 A protocol is presented for genetically engineering Cymbidium orchid using particle bombardment. This protocol enabled the routine transformation of orchid plants that were previously difficult to transform. Liquid culture was used to generate a large number of protocorm-like bodies (PLBs) to be bombarded and to promote continued development of the bombarded meristematic tissue. Plasmid DNA (pKH200) carrying the GUS-INT and NPTII genes flanked by tobacco matrix attachment regions was introduced into the meristematic cells of PLBs by particle acceleration. The transformed PLBs were proliferated and selected for kanamycin resistance conferred by the introduced NPTII gene. Shoot regeneration was then induced from the kanamycin-resistant PLBs, and transgenic plantlets were produced. Both the kanamycin-resistant PLBs and regenerated shoots expressed the GUS-INT gene. The presence of the introduced gene in the transformed orchid plants was confirmed by PCR analysis, sequencing and Southern blot analysis of the PCR product. The recovered transgenic plants were established in soil and acclimatized in the greenhouse. Received: 20 July 1998 / Revision received: 2 December 1998 / Accepted: 17 December 1998  相似文献   

15.
A cell suspension culture of Picea glauca (White spruce) which continuously produces somatic embryos has been established. Embryogenic callus derived from cultured zygotic embryos was used to initiate the culture. Numerous embryos at various early stages of development were recognized; they exhibited a meristematic embryonic region and suspensor consisting of elongate, vacuolated cells. The culture also contained clumps of meristematic cells and large irregular — shaped cells. The culture could be readily re-established on solid medium.  相似文献   

16.
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.  相似文献   

17.
Embryogenic calli were initiated from embryonic explants of Pinus roxburghii using female gametophytes containing immature pre-cotyledonary embryos. Zygotic embryos were collected at different developmental stages and cultured on various media. Initiation of embryogenic calli was achieved in pre-cotyledonary zygotic embryos of a 0.1-mm to 1.2-mm embryonal head on Douglus fir cotyledon revised medium (DCR) medium supplemented with 2,4-D or NAA and BA. Embryogenic callus development was initiated from the suspensor region of immature embryos. The method of immature embryo culture was significant as rapid embryogenic callus development occurred in megagametophytes where the suspensor was stretched onto the medium from the cut micropylar end. Sixty embryogenic lines were established from 2500 explants cultured during one season. A pro-embryo with six to eight meristematic cells and suspensor of six to ten long, vacuolated cells dominated the early phase of the callus development. Cleavage polyembryony occurred in proliferating callus, constituting a method of multiplication of these somatic embryos. Somatic embryos developed to stage-I and stage-II embryos on DCR medium supplemented with 5 μM 2,4-D or 10 μM NAA. Received: 30 June 1999 / Revision received: 15 November 1999 / Accepted: 3 December 1999  相似文献   

18.
文心兰原球茎液体增殖培养研究   总被引:11,自引:1,他引:10  
以茎尖诱导形成的原球茎(protocorm-like bodies,PLBs)为外殖体,采用液体培养方式比较了不同浓度的激素配比、蔗糖浓度和添加不同量的新鲜椰汁对文心兰PLBs增殖的影响,并比较了相同培养基成分时液体培养PLBs增殖、分化成苗和固体培养PLBs增殖和分化成苗的差异。试验结果表明:不同浓度的外源激素及其配比对文心兰PLBs增殖生长影响较大,6-BA0.5 mg/L Ad 0.05 mg/L NAA0.05 mg/L的激素组合比较适合文心兰PLBs增殖;蔗糖浓度对文心兰PLBs增殖的影响也较大,适合文心兰PLB在液体培养条件下增殖的蔗糖浓度为7.5 g/L;添加5%新鲜椰汁不仅对文心兰PLBs增殖有促进作用,而且能改善PLBs的质量;适合文心兰PLBs增殖的培养基为MS 6-BA0.5 mg/L Ad 0.05 mg/L NAA0.05 mg/L 5%椰汁 蔗糖7.5 g/L。文心兰PLBs在5周内的增殖生长曲线呈倒"V"字形,第3周的增殖速度达最高峰,而固体培养基PLBs增殖速度较慢,生长曲线几乎成直线。液体增殖的PLBs分化成苗较固体培养增殖的PLBs差。  相似文献   

19.
林荣  邹琦丽   《广西植物》1988,(1):89-91+105
用金桔茎段为外植体,培养在附加1.0毫克/升BA和0.l毫克/升IBA的MS培养基上,诱导愈伤组织和芽形成。观察了愈伤组织和芽形成过程中的组织细胞学变化。培养一周后,在茎组织切口两端开始膨大,细胞增大和开始分裂。培养两周后,开始形成瘤状愈伤组织。在愈伤组织中有形成层状分生组织、维管组织结节和分生细胞团。培养四周后,表层的分生细胞团分化形成大量芽原基,同时愈伤组织深层也出现分生细胞团。带节茎段可从切口两端的愈伤组织分化形成芽,亦可从叶腋的潜伏芽直接形成芽。  相似文献   

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