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1.
2,4-D、BA对人参体细胞胚胎发生过程的影响研究   总被引:1,自引:0,他引:1       下载免费PDF全文
以人参芽胞、二年生人参根、实生苗(茎、叶)为外植体研究了体细胞胚的发生条件,并对其发生过程中可溶性蛋白、相关酶活性及内源激素的变化等进行了研究。结果表明,诱导愈伤组织的培养基为MS+2,4-D4.0mg/L+BA0.2mg/L;在MS+2,4-D1.0mg/L+KT0.2mg/L培养基上继代培养,可获得胚性愈伤组织;在无2,4-D的培养基上可诱导出胚状体。将胚状体转入无任何激素的MS培养基上继续培养,之后转入1/2MS培养基上获得再生植株。在体细胞胚胎发生过程中,可溶性多糖和可溶性淀粉含量在早期胚时较低,可溶性蛋白含量、POD及PPO活性在早期胚时最高;IAA在早期胚时期含量最高,在成熟胚时期ABA含量最高,而ABA/IAA比值在成熟胚时较高,利于体细胞胚的发育成熟。  相似文献   

2.
‘SK4—316’胡萝卜体胚的诱导和培养   总被引:2,自引:0,他引:2  
以'SK4-316'胡萝卜无菌苗的下胚轴为外植体,研究不同培养基配方和培养条件对愈伤组织诱导、体细胞胚间接发生及其同步化培养的影响,以及不同脱分化时间、脱分化培养基及外植体续存时间对体细胞胚直接发生的诱导及其培养的影响.结果表明:含3%蔗糖、0.8%琼脂的1/2MS + 2,4-D 2.5 mg/L + 6-BA(或KT)0.5 mg/L + CH 300 mg/L是诱导愈伤组织的良好培养基;1/2MS + 2,4-D 1.25 mg/L + KT 0.25 mg/L + 6-BA 0.25 mg/L(含3%蔗糖)适于愈伤组织分化并诱导体胚发生,0.02% ABA对体胚的诱导有促进作用,0.06% ABA或15% PEG能促进体胚成熟;外植体在MS + 2,4-D 1.0 mg/L固体培养基上脱分化培养48 h,再转入MS + CH 300 g/L液体培养基中可诱导体胚直接发生,但随着外植体续存于诱导培养基中时间的延长,体胚发生变异的几率也渐增.  相似文献   

3.
云南大叶茶体细胞胚发生及体细胞胚苗形成体系的建立   总被引:9,自引:0,他引:9  
利用云南大叶茶(Camellia sinensis var.assamica Kitamura)胚性细胞系(CL_1)中悬浮培养物,建立了高频率同步化体细胞胚发生及体胚苗形成体系。以改良的MS为基本培养基,将CL_1中培养物由液体保持培养基(0.1mg/L 2,4-D 0.5mg/L 6-BA)继代转入液体诱导培养基(0.05mg/L 2,4-D 0.50mg/L6-BA),暗培养诱导28d,转入不含任何激素的液体分化培养基中再培养28d,获得了不同发育时期的体细胞胚,其发生频率为81.5%。不同发育时期的体细胞胚用不同目的细胞筛收集,在液体生长培养基(1/2 MS 1.0mg/L GA_3 0.5mg/L 6-BA)中培养发育成熟。ABA有利于高质量体细胞胚的形成。20~70月大小的体细胞胚在固体生长培养基中成苗转换率为75%。在液体悬浮培养条件下观察记录了体细胞胚发育过程,证实其过程与合子胚的形态发生过程相似。  相似文献   

4.
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。  相似文献   

5.
三七胚培养中的胚胎发生   总被引:5,自引:0,他引:5  
三七成熟胚培养于MS 1 mg/l IAA或NAA或2,4-D的培养基上。二月后,在MS 1mg/l IAA或NAA的培养基上由外植体可诱导产生胚状体,但在含2,4-D的培养基上只产生愈伤组织而无器官分化。胚状体转入MS GA_3 1mg/l IAA0.5 mg/l培养基上可发育成具胚根、根芽的小植株。  相似文献   

6.
ABA诱导毛酸浆下胚轴体细胞胚状结构的直接发生   总被引:3,自引:0,他引:3  
研究了ABA与2,4-D等植物激素对毛酸浆组织培养的影响。其中,以浓度为1.0mg/L的2,4-D对毛酸浆下胚轴愈伤组织诱导结果最好。在子叶、子叶柄、下胚轴以及胚根四者当中,下胚轴对2,4-D的反应状况最好。结合ABA与2,4-D在MS培养基上能够有效地诱导出高质量的球形胚状结构。经过4周的培养,在2.0mg/L2,4-D和1.0mg/L ABA组合的情况下,球形胚状结构的诱导频率可达96.7%,每块外植体平均能产生6.7个球形胚状结构。球形胚状结构可以至少在5代以内反复发生。进一步讨论了ABA对毛酸浆体细胞胚发生的影响以及体细胞胚发生与环境胁迫的可能关系。  相似文献   

7.
防风组织培养中畸形胚状体的发生和控制   总被引:10,自引:0,他引:10  
采用组织培养和石蜡切片法,分析多种因素对防风畸形胚状体发生和发育过程的影响及控制。结果表明,诱导正常体细胞胚高频发生的培养基组合是:启动胚性愈伤组织的培养基是MS 0.5mg/L 2.4-D,蔗糖浓度3%;分化培养基是MS 1mg/L 6-BA 0.2mg/L NAA 0.5mg/L ABA,蔗糖浓度4%~5%;成熟培养基是MS培养基,蔗糖浓度3%。结论:一定浓度的生长素是诱导防风胚性愈伤组织的关键因素,细胞分裂素在体细胞胚的分化和发育过程中起协同作用;蔗糖浓度、ABA、接种方法以及适宜的培养条件和培养容器等均可有效降低畸形胚状体的发生率。  相似文献   

8.
枸杞悬浮培养条件下的胚状体发生   总被引:5,自引:0,他引:5       下载免费PDF全文
在MS培养基上诱导宁夏枸杞下胚轴切段形成愈伤组织,并进行细胞悬浮培养。观察了悬浮培养条件下胚状体的发生过程。<1>观察发现,细胞经悬浮培养几天以后,先形成胚性细胞团,再有胚性细胞团块形成一个或几个胚状体。较大的胚状体也可以形成新的次生胚状体。<1>影响胚状体形成的关键因素是激素的种类及其含量。实验采用的基本培养基为MS,比较适宜胚状体发生的激素是0.2mg/L 2,4-D。同样浓度的2,4-D,会抑制胚状体的进一步发育,用0.2mg/L6-BA代替2,4-D, 则胚状体可以进一步发育并形成小植株。  相似文献   

9.
黄瓜成熟胚离体培养中的胚状体诱导和植株再生(简报)   总被引:11,自引:0,他引:11  
以10个黄瓜品种为材料,取成熟胚外植体先经MS 2,4—D 1.0mg/L(单位下同) KT0.5诱导后转入MS IAA 0.1 KT0.5上分化,胚状体发生频率高并有植株再生。材料的基因型差异显著,“农大春光”分化率最高(40%)。双层培养有效地促进畸形胚状体的正常发育,5%蔗糖和1.0mg/L ABA促进胚性愈伤组织的继代增殖和胚性保持。  相似文献   

10.
三七为五加科的重要药用植物。以三七的种胚为材料接种于MS 补加2,4-D、IAA、NAA 各1 mg/L的培养基上。接种后约二个月,在MS 补加IAA 或NAA 的培养基上可以产生体细胞胚;在MS 加2,4-D 的培养基上只产生愈伤组织而无器官分化。体细胞胚在MS+IAA 0.5 mg/L+GA_3 1 mg/L 培养基上可发育成小植株。体细胞胚用4%海藻酸钠和2%氯化钙进行人工种皮包埋后,在无菌条件下,人工种子可转换成苗,转换率达89.7%。三七的体细胞胚起源于愈伤组织内或近表层的单个胚性细胞。体细胞胚经球形、心型、鱼雷形及子叶期等诸阶段发育成植株。通过PAS 法染色发现,胚性细胞、球形胚、早心形胚阶段有明显地淀粉积累,淀粉颗粒大而密集;到晚心形胚及子叶期体胚,淀粉颗粒小而少或消失。淀粉消长变化的规律与体细胞胚的发育有着密切的关系。  相似文献   

11.
Summary Immature embryos of 41 lines of barley were screened in vitro for callus induction and somatic embryogenesis on different media to establish totipotent cultures. The use of modified MS and CC media, both supplemented with 1 g/l casein hydrolysate, and the substitution of agarose for agar resulted in the highest frequencies of somatic embryo induction. Embryogenic callus was induced and plants regenerated from 23 of the lines tested. The auxins 2,4-D, dicamba, picloram and 2,4,5-T were suitable for embryogenic callus induction. High frequencies of somatic embryo germination occurred on CC medium supplemented with 1 mg/l IAA and 0.05 mg/l zeatin. A strong genotypic effect on the capacity and frequency of embryogenic callus formation was found. Cultivar Golden Promise always gave the best results. Experiments with field grown material in 3 consecutive years showed that environmental factors also strongly influenced the induction of somatic embryogenesis and plant regeneration.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - 2,4,5-T 2,4,5-trichlorophenoxyacetic acid - dicamba 3,6-dichloro-o-anisic acid - picloram 4-amino-3,6,6-trichloropicolinic acid - NAA naphtaleneacetic acid - IAA indole-3-acetic acid - ABA abscisic acid - BAP 6-benzyl amino purine - 2iP 6-(3-methyl-2 butenyl 1-amino)purine - GA3 gibberellic acid  相似文献   

12.
A procedure for rapid multiplication of Chrysanthemum morifolium RAMAT cv. Birbal Sahni using leaf callus and stem (nodal/internodal) callus as well as node and apical shoots has been developed. Murashige and Skoog's medium (1962) supplemented with 2mg/1 2,4-D yielded good green calli from both leaf and stem segments within 2 weeks. About 1 cm × 1 cm callus regenerated 2–3 shoots after 3 weeks on MS solid medium supplemented with 0.1 mg/l IAA and 0.2 mg/l BAP. Each of the regenerated shoots when transferred to the same shooting medium without agar yielded about 150 new shoots, which in turn regenerated roots after another week in MS half strength or modified White's media (Rangaswamy, 1961). It has been estimated that about 1014 plantlets could be produced in a year from one expiant following the proposed protocol.Abbreviations BAP 6-benzylaminopurine - IAA indole-3-acetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - MS Murashige and Skoog's (1962) medium  相似文献   

13.
We investigated the optimal levels of growth regulators, culture media, and pH on callus growth and organogenesis of in-vitro cultured ‘Kyoho’ grapes. Calli were induced by culturing leaf blades on an MS basal medium supplemented with 1 mg/IL BA and 0.01 mg/L 2,4-D. In addition, calli originating from the exocarp and mesocarp of grape fruits devel-oped on MS media supplemented with 0.1 mg/L IAA, NAA, or 2,4-D, or with 0.2 mg/L BA. In testing the potential for plant regeneration from shoot tips on various media, we found that the Nitsch medium, with I mg/L BA, was optimal for caulogenesis. The type of shoot development depended on the pH of the medium, with vigorous multiple-shoot devel-opment occurring at pH 6.0, and single shoots forming at pH 5.0. Finally, we were able to obtain rooted seedlings from the regenerated shoots that had been cultured on 1/4-strength Nitsch medium supplemented with 0.03 mg/L NAA.  相似文献   

14.
In order to investigate the effect of ABA on secondary embryogenesis from somatic embryos inAralia cordata Thunb., embryogenic callus and somatic embryos were induced from inflorescence on solid MS basal medium supplemented with 1.5 mg/L 2,4-D after eight weeks without subculture. For mass production of somatic embryos, embryogenic cell clumps were maintained in liquid MS medium supplemented with 1.0 mg/L 2,4-D, and then transferred to 2, 4-D-free medium. When developing embryos at various stages were cultured separately in liquid medium with ABA (0 to 2.0 mg/L) for three weeks, and then cultured in ABA-free liquid medium for two weeks, torpedo-shaped embryos exhibited secondary embryogenesis of 65.9% in only 0.2 mg/L ABA pretreatment. Cotyledonary embryos in cultures by 0.2, 0.5 and 1.0 mg/L ABA pretreatment also exhibited secondary embryogenesis (73%, 9.4% and 6.0%, respectively). However, globular and heart-shaped somatic embryos treated with ABA did not form secondary embryos on their hypocotyl surfaces. When cotyledonary embryos were cultured in ABA-free medium or 0.2 mg/L ABA treated medium for three weeks, and then in ABA-free liquid medium for 6 weeks, the germination frequency was lower in medium with 0.2 mg/L ABA (45.9%) than in hormone-free medium (56.8%). This result seems to be related to the high frequency of secondary embryogenesis. It is suggested that secondary embryogenesis by ABA application depends upon the stage of embryo cultured and the ABA concentration.  相似文献   

15.
目前转基因技术已成为植物定向遗传改良的重要手段,而建立稳定高频的离体再生系统是实现遗传转化的基础和前提.本试验以25 ~30 d苗龄的金养麦(Fagopyrum dibotrys)无菌苗叶片、茎节间、叶柄为外植体进行愈伤组织诱导与植株再生研究.结果表明:叶片在MS +2,4-D 4.0 mg/L +6-BA 1.0 mg/L培养基上愈伤组织诱导率达到89%.茎节间在MS +2,4-D 2.0 mg/L +6-BA 2.0 mg/L培养基上愈伤组织诱导率为87%.叶柄在MS +2,4-D 4.0 mg/L +6-BA 2.0 mg/L+ IBA 0.2 mg/L培养基上的最高诱导率仅为54%.愈伤组织分化不定芽的适宜培养基为MS +6- BA2.0 mg/L +TDZ0.2 mg/L +NAA0.2 mg/L;金荞麦不定芽在1/2 MS +NAA 0.5 mg/L的培养基上生根效果最好.组培再生植株经炼苗后移栽到田间成活率达80%以上,且生长表现正常.高频完整再生体系的建立,为金荞麦进一步遗传操作和扩大药材资源奠定了基础.  相似文献   

16.
新疆天山雪莲体胚诱导与分化研究   总被引:5,自引:0,他引:5  
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.  相似文献   

17.
三七成熟胚培养子MS+1 mg/L 1AA或NAA或2,4 — D的培养基上。二月后,在MS+1 mg/L 1AA或NAA的培养基上由外植体可诱导产生胚状体,但在含2,4 — D的培养基上只产生愈伤组织而无器官分化。胚状体转入MS+CA3 1 mg/L+IAAo.5 mg/L培养基上可发育成具胚根、根芽的小植株。  相似文献   

18.
Somatic embryos were obtained from callus cultures derived from leaf explants of the winged bean, Psophocarpus tetragonolobus (L.) DC. Initiation and development of the somatic embryos occurred with a two-step culture method. Callus cultures initiated on MS medium with NAA and BAP, upon transfer to a new medium with IAA and BAP, produced somatic embryos. Maximum embryogenesis of 60% was obtained on induction medium with 0.5 mg/l NAA plus 1.0 mg/l BAP followed by transfer to a secondary medium with 0.1 mg/l IAA and 2.0 mg/l BAP. Optimal embryo germination and plantlet development was achieved on MS medium with 0.2 mg/l BAP plus 0.1 mg/l IBA. The regenerated plants were successfully transferred to glasshouse conditions.Abbreviations MS Murashige and Skoog (1962) medium - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - IAA Indole-3-acetic acid - IBA Indole-3-butyric acid - BAP 6-benzylaminopurine - KN Kinetin  相似文献   

19.
Embryogenic callus and somatic embryos were induced from cotyledonary explants of African marigold (Tagetes erecta L.). Cotyledons were first cultured on MS medium supplemented with 2.0 mg l–1 2,4-D and 0.2 mg l–1 kinetin. After 5 weeks, calli were transferred to MS medium supplemented with 0.02 mg l–1 thidiazuron where compact embryogenic callus developed. Friable embryogenic callus developed when the compact embryogenic callus was transferred to medium containing 2,4-D and subcultured every 2 weeks. Friable embryogenic callus has been maintained for more than 2 years without losing the capacity to generate embryos. Embryo development was obtained when friable embryogenic callus was transferred to MS medium supplemented with 3 mg l–1 ABA and 60 g l–1 sucrose. The addition of 10–30 mM l-glutamine improved embryo development. Received: 13 May 1997 / Revision received: 24 February 1998 / Accepted: 28 March 1998  相似文献   

20.
Mature endosperm of Emblica Officinalis (Euphorbiaceae) formed a continously growing callus on MS medium supplemented with an auxin (2,4-D or IAA) and a cytokinin (K or BAP). Subculture of callus on MS with BAP (0.2 mg/l) and IAA (0.1 mg/l) resulted in formation of shoots and embryo-like structures in 50 and 8 per cent cultures, respectively. Regeneration of shoots was more frequent when both BAP (0.2 mg/l) and IAA (0.1 mg/l) were present than on BAP (0.2 mg/l) alone. The embryo-like structures produced plantlets.Abbreviations BAP benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - GA3 gibberellic acid - IAA indoleacetic acid - K kinetin - NAA naphthaleneacetic acid - PDB para-dichlorobenzene (née Arora)  相似文献   

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