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1.
从印度娃儿藤节间外植体获取愈伤组织,分析了糖、赤霉素(GA3)及脱落酸(ABA)对愈伤组织形成体细胞的影响。实验证明,含4 μmol/L 2, 4-二氯苯氧乙酸(2, 4-D)的MS培养基是获得具有成胚功能的愈伤组织的最佳培养基。在含有6 μmol/L激动素(Kn)的MS培养基上,高达69%的愈伤组织分化为体细胞胚,平均单位外植体(每克愈伤组织)产胚25个。在6 μmol/L Kn存在的条件下,分析了蔗糖、葡糖糖对胚产生的影响,不同的糖及不同糖浓度对体细胞胚的发生影响很大。6 μmol/L Kn 与200 mmol/L 蔗糖处理胚胎发生率最大(71%),单位外植体生成49个胚。然而葡萄糖与Kn、或者葡糖糖、蔗糖与Kn三者加在一起则降低成胚率及产胚数。一定浓度的GA3 和 ABA能促进体细胞胚的产生。在含200 mmol/L蔗糖的培养基中加10 μmol/L GA3胚的生成率为98%,单位外植体产胚51个。在含200 mmol/L蔗糖的培养基中加2 μmol/L ABA能显著增加体细胞胚的量,该培养基上每外植体平均生成44个胚,产率为95%。本研究显示,含200 mmol/L蔗糖的培养基中分别加入6 μmol/L Kn、10 μmol/L GA3 或者 2 μmol/L ABA能显著提高印度娃儿藤体细胞胚发生率,而单独的葡萄糖或葡糖糖和蔗糖则有抑制作用。得到的胚均能正常发育并分化为植株。  相似文献   

2.
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   

3.
‘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   

4.
本文以秦艽叶片和茎段作为外植体,通过离体培养对秦艽植株再生途径进行研究。愈伤组织在添加2mg/L 2,4-D和0.5mg/L BA的MS培养基上诱导,两周内可出现愈伤组织。愈伤组织在相同激素配比并附加500mg/L LH的MS培养基上继代。愈伤组织的分化在添加有0.1mg/L 2,4-D和0.5mg/L BA的MB培养基上进行。通过显微观测,疑似体细胞胚可以在叶片和茎段的愈伤组织上产生。形态学和组织学的分析进一步证实了秦艽离体再生过程中体细胞胚发生的现象。体细胞胚和合子胚一样,也经历球形、心形、鱼雷和子叶胚等发育时期。相对独立的结构说明秦艽的体细胞胚可能是单细胞来源。体细胞胚在愈伤组织的表面和内部都有出现。在本实验中,体细胞胚发生途径是在秦艽愈伤组织形成后观察到的唯一再生途径。  相似文献   

5.
白杄体细胞胚胎发生及其植株再生   总被引:9,自引:0,他引:9  
白杄(Picea meyeri Rehd.et Wils.)的成熟胚在4~6℃低温下保存1个月后,接种于改良LP 2mg/L 2,4-D 1mg/L 6-BA的培养基上,黑暗条件下培养1个月便可产生白色半透明的胚性愈伤组织。整体染色封片观察表明,胚性愈伤组织由很多很长的胚柄细胞及其顶端的胚细胞团组成,这种愈伤组织培养物称为胚性胚柄团。胚性胚柄团在MS 1mg/L 2,4-D 1mg/L KT的继代培养基上黑暗条件下可保持旺盛的增殖能力和分化潜力。当胚性胚柄团转到MS 5mg/L ABA 5mg/L AgNO_3的分化培养基上,1个月后可产生大量正常的体细胞胚。体细胞胚成熟以后转到含0.5%活性炭的无激素1/2MS基本培养基上约40d后可长出1.5~2.5cm的根,约60d后可长出真叶。光、ABA、蔗糖及AgNO_3浓度是影响体细胞胚发生的主要因素。  相似文献   

6.
影响籼稻体细胞胚胎发生几个因素的研究   总被引:13,自引:0,他引:13  
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。  相似文献   

7.
用大豆(Glycine max(L.)Merr.)未成熟胚为外植体,在含高浓度 NAA 或2,4-D 的培养基上可诱导绿色结构体和体细胞胚状体。叶状、花状和喇叭形状的绿色结构体在含30mg/l2,4-D 的 MS-1培养基上可重新愈伤组织化。这种由高浓度2,4-D 诱发并保持的愈伤组织,转到低激素水平的培养基上后,能诱导出大量的同类形状的新结构体。绿色结构体可以进一步发育成叶丛状,但顶端不伸长。使用含2m g/l GA,和0.1mg/l IBA 的培养基,能促进顶端伸长和根的发育,获得正常植株。由未成熟胚的子叶(其时种子长度约4—6mm)诱导的愈伤组织具有再生能力。在含高浓度2,4-D(5-30mg/l)的培养基上诱导出的愈伤组织具有较高的产生绿色结构的潜力。而高浓度 NAA(10mg/l)培养基产生的愈伤组织具有较强的产生根状体的潜力。培养的外植体或愈伤组织在含5mg/l 2,4-D 的培养基上,不需要经常的继代培养,即可长期保存。  相似文献   

8.
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   

9.
小麦幼穗胚性愈伤组织诱导及分化过程中内源激素的作用   总被引:19,自引:2,他引:17  
2,4-D的浓度及诱导时间的长短影响出愈率和早期体胚的形成,6-BA0.25mg/L或KT0.5mg/L抑制胚性愈伤组织的诱导及分化。胚性能力较强外植体的内源激素含量较高,同时分化过程中胚性愈伤组织的ABA、IAA含量高于非胚性意伤组织,而GA3含量则低于非胚性愈伤组织。  相似文献   

10.
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   

11.
Summary A procedure for the regeneration of cacao (Theobroma cacao) plants from staminode explants via somatic embryogenesis was developed. Rapidly growing calli were induced by culturing staminode explants on a DKW salts-based primary callus growth (PCG) medium supplemented with 20 g glucose per L, 9 μM 2,4-D, and thidiazuron (TDZ) at various concentrations. Calli were subcultured onto a WPM salts-based secondary callus growth medium supplemented with 20 g glucose per L, 9 μM 2,4-D, and 1.4 nM kinetin. Somatic embryos were formed from embryogenic calli following transfer to a hormone-free DKW salts-based embryo development medium containing sucrose. The concentration of TDZ used in PCG medium significantly affected the rate of callus growth, the frequency of embryogenesis, and the number of somatic embryos produced from each responsive explant. A TDZ concentration of 22.7 nM was found to be the optimal concentration for effective induction of somatic embryos from various cacao genotypes. Using this procedure, we recovered somatic embryos from all 19 tested cacao genotypes, representing three major genetic group types. However, among these genotypes, a wide range of variation was observed in both the frequency of embryogenesis, which ranged from 1 to 100%, and the average number of somatic embryos produced from each responsive explant, which ranged from 2 to 46. Two types of somatic embryos were identified on the basis of their visual appearance and growth behavior. A large number of cacao plants have been regenerated from somatic embryos and established in soil in a greenhouse. Plants showed morphological and growth characteristics similar to those of seed-derived plants. The described procedure may allow for the practical use of somatic embryogenesis for clonal propagation of elite cacao clones and other applications that require the production of a large number of plants from limited source materials.  相似文献   

12.
Efficient plant regeneration through somatic embryogenesis was achieved from callus cultures derived from semi-mature cotyledon explants of Dalbergia sissoo Roxb., a timber-yielding leguminous tree. Somatic embryos developed over the surface of embryogenic callus and occasionally, directly from cotyledon explants without intervening callus phase. Callus cultures were initiated from cotyledon pieces of D. sissoo on Murashige and Skoog (1962) medium supplemented with 4.52, 9.04, 13.57, and 18.09 mumol/L 2,4-dichlorophenoxyacetic acid and 0.46 mumol/L Kinetin. Maximum percentage response for callus formation was 89% on MS medium supplemented with 9.04 mumol/L 2,4-D' and 0.46 mumol/L Kn. Somatic embryogenesis was achieved after transfer of embryogenic callus clumps to 1/2-MS medium without plant growth regulators (1/2-MSO). Average numbers of somatic embryos per callus clump was 26.5 on 1/2-MSO medium after 15 weeks of culture. Addition of 0.68 mmol/L L-glutamine to 1/2-MSO medium enhanced somatic embryogenesis frequency from 55% to 66% and the number of somatic embryos per callus clump from 26.5 to 31.1. Histological studies were carried out to observe various developmental stages of somatic embryos. About 50% of somatic embryos converted into plantlets on 1/2-MSO medium containing 2% sucrose, after 20 days of culture. Transfer of somatic embryos to 1/29-MSO medium containing 10% sucrose for 15 days prior to transfer on 1/2-MS medium with 2% sucrose enhanced the conversion of somatic embryos into plantlets from 50 to 75%. The plantlets with shoots and roots were transferred to 1/2 and 1/4-liquid MS medium, each for 10 days, and then to plastic pots containing autoclaved peat moss and compost mixture (1:1). 70% of the plantiets survived after 10 weeks of transfer to pots. 120 regenerated plantlets out of 150 were successfully acclimatised. After successful acclimatisation, plants were transferred to earthen pots.  相似文献   

13.
Direct somatic embryo induction was achieved from leaf and internodal explants of Solanum tuberosum (L.) cultivar ‘Kufri Chipsona 2’ on Murashige and Skoog (Physiol Plant 15:473–497, 1962) medium containing 10.0 µM silver nitrate (MS1 medium) supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D; 2.5 µM) and 6-benzyladenine (BA; 1.0 µΜ). It was observed that in absence of AgNO3, friable callus was induced from cut ends of the explants, which does not develop into any kind of organised structure; thus highlighting the requirement of AgNO3 for somatic embryogenesis in potato. Furthermore, the effect of medium strength, sucrose concentration and heat shock treatment on somatic embryogenic potential of explants was also investigated. When the strength of basal medium was reduced to half, the frequency of internodal segments differentiating somatic embryos was almost double in comparison to full strength MS medium. Sucrose concentration and heat shock treatment were found to have interactive effect on somatic embryo induction. Explants subcultured on medium containing 174 mM sucrose and subjected to heat shock (1 h; 50 °C) showed maximum somatic embryo differentiation. Although, the percent explants differentiating somatic embryos decreased sharply with increase in sucrose concentration (>?174 mM), yet the number of somatic embryos differentiated per explant were found to increase with further increase in sucrose concentration. Histological observations revealed that somatic embryos directly developed from epidermis of leaf explant and cut ends of internodal segments progressed from globular to cotyledonary stage after passing through intermediate embryogenic stages (heart shaped and torpedo shaped). Conversion of somatic embryos into plantlets (92%) was achieved on MS1 medium supplemented with BA (10.0 µM) and gibberellic acid (15.0 µM) and all regenerated plants were found to be phenotypically alike.  相似文献   

14.
贡蕉胚性细胞悬浮系的建立和植株再生   总被引:21,自引:0,他引:21  
鲜食蕉品种的高度不育性和多倍性制约了用传统育种方法培育生产实践中所需的新品种 ,建立稳定的胚性细胞悬浮系是香蕉生物技术育种的前提。以目前国内尚未建立该体系的鲜食蕉品种贡蕉 (AA)未成熟雄花序的第 1~ 15位花梳为外植体 ,对胚性细胞悬浮系的建立和植株再生体系进行了优化。结果表明 ,5~ 6个月的培养后可获得分生小球体和浅黄色、松散易碎的胚性愈伤组织。 9μmol/L 2,4 D对外植体愈伤组织的诱导效果最好 ,诱导率为 40.96 % ,胚性愈伤组织诱导率可达7.45 % ,其中5.79%的胚性愈伤组织来源于第 6~12号位置的花梳。胚性愈伤组织悬浮培养后 ,通过 3个月的筛选和继代培养 ,可得到均质的胚性细胞悬浮系。该培养体系合适继代周期为 15d ,继代时合适的起始接种量为每 30mL培养基加 2mLPCVECS。培养 6个月的胚性细胞在体细胞胚诱导培养基中培养15d后可见到白色半透明体细胞胚的发生 ,体细胞胚诱导率为 2 80× 103个 mLPCV。成熟体细胞胚的萌发率为 17 2 8% ,其中发育成正常的再生植株的百分率为 14 16 %。  相似文献   

15.
Brassinolide (BR), which is the most biologically active brassinosteroid, was used to examine the potential effect of hormone on cotton somatic embryogenesis. Ten-day-old cotton (Gossypium hirsutum L., cv. Cooker) seedlings were used for explant source and hypocotyls were removed and cultured on MS basal medium with B5 vitamins supplemented with 1 mg/L 6-benzylaminopurine + 0.5 mg/L kinetin for callus induction. After one month proliferating calli pieces were collected and cultured on MS basal medium containing various concentrations of BR (0.1, 0.5, 1.0 μM) with their controls. BR treatments were negatively effective on the fresh weight of calli when compared to control. Differential somatic embryogenesis maturation rates due to BR treatment were observed. Somatic embryogenesis was stimulated especially for transition to cotyledonary phase at 0.5 mg/L BR. Histological preparations from embryogenic calli and somatic embryos at different stages of development revealed the spontaneous polyploidisation during early somatic embryogenesis on BR-treated calli. Present results suggest that BR negatively effected calli growth, however, had a stimulating role in maturation of somatic embryos.  相似文献   

16.
An efficient in vitro plant regeneration system was established through somatic embryogenesis for Anoectochilus elatus Lindley, an endangered jewel orchid. Direct somatic embryogenesis was achieved from nodal explants (17.4 embryos per explant with 63.4% response) on Mitra medium supplemented with Morel vitamins, thidiazuron (4.54 µM) and ∞-naphthaleneacetic acid (2.69 µM). Simultaneously, a protocol was developed for indirect somatic embryogenesis from internodal explant, produced embryogenic calli and embryos (31.3 embryos with 76.4% response) on same medium amended with 50 mg/L peptone and 5% coconut water. Both types of embryogenic pathways, produced morphologically similar globular embryos in the form of protocorm like bodies and successfully germinated on hormone free Mitra medium supplemented with Morel vitamins. Morpho-histological investigation of the embryo revealed the initiation and developmental features of somatic embryos. In vitro regenerated plantlets were successfully established from heterotrophic to a photoautotrophic stage by reducing the nutrient content in culture media, adjusting temperature and humidity through three step method. During the process, no morphological and physiological abnormalities were observed. Hardened plantlets were successfully acclimatized at poly tunnel chamber with 95% of survival rate. Further, inter simple sequence repeats (ISSRs) molecular markers were used to analyse the genetic homogeneity of regenerated plants. Analysis with this method showed that the homogeneity is comparatively higher in direct somatic embryo regenerated plants (94.22%) as compared to plants elevated from an indirect somatic embryo (93.05%). The present study provides morpho-histological and genetically stable plants for germplasm conservation and further utility of this endangered jewel orchid.  相似文献   

17.
The system of high synchronous frequency of somatic embryogenesis and somatic embryo seedling formation was established by means of embryonic cell hne 1 ( CL1 ) of Camellia sinensis var. assamica Kitamura. Modified MS was used as the basic medium. Cultures of CL1 was transferred to the aqueous induced medium (0.05 mg/L 2,4-D + 0.50 mg/L 6-BA) from the maintenance medium (0.1 mg/L 2,4-D + 0.5 mg/L 6-BA) for somatic embryos induction under dark condition. 28 days later, they were cultured in the liquid differentiation medium. Various kinds of somatic embryos were obtained after another 28 days. The frequency of somatic embryos was 81.5 %. Various mesh sizes of sieves were applied to collect the somatic embryos in different developmental stages which could develop to mature stage in the aqueous growth medium ( 1/2 MS + 1.0 mg/L GA3 + 0.5 mg/L 6-BA). ABA was effective to promote the formation of highly qualified somatic embryo. The mature somatic embryos sized 20 to 70 mesh had the conversion frequency 75 %. The development of somatic embryogenesis studied under a cell suspension culture system was similar to the zygotic embryogenesis.  相似文献   

18.
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   

19.
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   

20.
白Pian体细胞胚悬浮培养的动力学研究   总被引:1,自引:0,他引:1  
白(PiceameyeriRehd.etWils.)是我国特有的云杉属树种,在林业生产和环境绿化中均具有重要地位。其体细胞胚胎发生的研究,一方面可用于优良种质的大规模快速繁殖,为植树造林和园林绿化提供优质苗木;另一方面可作为遗传转化的再生系统,进行树种遗传...  相似文献   

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