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1.
莴苣子叶培养中RNA,蛋白质及过氧化物酶的变化   总被引:2,自引:1,他引:1  
莴苣子叶在含BA2mg/L(单位下同)和NAA0.2或BA0.2和NAA2的培养基中,均能形成胚性愈伤组织,在2,4-D2中形成非胚性愈伤组织。前一组合的RNA、蛋白质含量及过氧化物酶活性明显大于后一组合,过氧化物酶同工酶谱谱带宽且色深,具有一条特异酶带。由BA和NAA两种组合培养基诱导的胚性愈伤组织的三种生化指标变化的总趋势相同,同工酶谱相似,高BA组比低BA组的生化指标略高些。  相似文献   

2.
在 LS 附加1mg/1 BA+1mg/l KT 的培养基上,红豆草(Onobrychis viciaefolia Scop.)无菌苗的下胚轴切段产生淡黄色的愈伤组织。愈伤组织转移到 LS 附加1mg/l BA 的培养基上,诱导体细胞胚胎发生,而在 LS 附加1mg/l KT 的培养基上抑制体细胞胚胎发生。同时,发现红豆草胚性愈伤组织中游离脯氨酸的含量仅为非胚性愈伤组织的2/5。向培养基中加入L-脯氨酸可以促进红豆草体细胞胚胎发生。最适浓度为1000mg/l。  相似文献   

3.
魏琴  曹有龙  陈放  周黎军  陈东林   《广西植物》2000,20(2):168-171+203
枸杞髓组织在 MS+6 - BA0 .1mg/ L+NAA0 .5mg/ L培养基上诱导愈伤组织发生。在 MS+6 - BA0 .1mg/ L+NAA0 .5mg/ L+CH50 0 mg/ L培养基上继代培养 ,再转入 MS+6 - BA2 mg/L +NAA 0 .5mg/ L的分化培养基上进行分化培养。显微观察表明 ,在培养过程中愈伤组织细胞由非胚性细胞转变为胚性细胞 ,直至发育成体细胞胚胎和完整植株 ;电泳结果显示 ,体细胞胚胎发生的各阶段 ,其过氧化物酶同工酶发生相应的变化。  相似文献   

4.
通过转移洋桔梗非胚性愈伤组织到含有1.0mg/L2,4-D的MS培养基(ecIM)上诱导了洋桔梗胚性愈伤组织形成,而非胚性愈伤组织在含1.0mg/L2,4-D和0.5mg/LKT的MS培养基(necSM)上继代培养。本研究比较分析了洋桔梗愈伤组织在ecIM和necSM上的超氧化物歧化酶(SOD)活性及其同工酶酶谱、酯酶同工酶酶谱随着培养天数的变化。实验结果表明在ecIM和necSM上培养的洋桔梗愈伤组织的超氧化物歧化酶(SOD)活性在培养早期较低,然后随着培养天数增加而升高,维持在较高水平上,但SOD活性变化无明显规律性;另一方面,SOD同工酶在第4天后出现一新的同工酶谱带;此外,在ecIM和necSM上培养洋桔梗愈伤组织的酯酶(EST)同工酶在培养至第16~20天期间呈现显著缺失。  相似文献   

5.
楸树体细胞胚胎发生过程中4种同工酶分析   总被引:1,自引:0,他引:1  
采用不连续聚丙烯酰胺凝胶电泳技术对楸树体细胞胚胎发生过程中的酯酶(EST)、过氧化物酶(POD)、淀粉酶(AMY)及ATP酶4种同工酶进行分析。结果表明,EST及POD同工酶酶带在楸树体细胞胚胎发生不同时期呈现规律性变化,胚性愈伤组织中EST、POD同工酶酶带较非胚性愈伤组织多且表达活跃,子叶胚时期活性最强,表明这一时期细胞内代谢旺盛。EST、POD、AMY及ATP同工酶在楸树胚性与非胚性愈伤组织中谱带差异明显,表明这4种同工酶与体胚发生具有密切关系,可以作为楸树胚性愈伤组织和体胚发生的重要标志。  相似文献   

6.
本文报道石刁柏胚性愈伤组织的可溶性蛋白质含量与组分、过氧化物酶和酯酶的活力及同工酶带均比其体细胞胚的要少。而在体细胞胚胎发生过程中,过氧化物酶和酯酶活力、可溶性蛋白质含量均以球形胚为最低,子叶分化期胚为最高而呈递增趋势;可溶性蛋白质组分以子叶分化期胚、成熟胚为最多,球形胚、香蕉形胚为最少;过氧化物酶同工酶带以梨形胚为最多,子叶分化期胚、成熟胚为最少;酯酶同工酶则以子叶分化期胚为最多,成熟胚为最少。胚性愈伤组织与体细胞胚均有其特异性可溶性蛋白质及同工酶带,可作为体细胞胚胎发生的分子标记。  相似文献   

7.
香果树体细胞胚胎发生过程中4种同工酶的研究   总被引:5,自引:1,他引:4  
用非变性聚丙烯凝胶电泳技术对香果树体细胞胚胎发生及形态建成过程中过氧化物酶(POD)、酯酶(EST)、淀粉酶(AMY)和超氧化物歧化酶(SOD)4种同工酶进行分析.结果表明:香果树体细胞胚胎发生及形态建成过程中,POD、EST、AMY和SOD活性变化与胚性愈伤组织的诱导及体细胞胚的发生发育密切相关.非胚性愈伤组织和胚性愈伤组织酶谱差异明显,胚性愈伤组织中EST和AMY同工酶酶带多且活性高,非胚性愈伤组织中缺乏EST和AMY同工酶表达,AMY同工酶可作为胚性细胞分化和发育的重要标志.香果树体细胞胚形态建成过程中,球形胚时期的AMY、POD、EST同_T酶活性最强,表明这一时期生理代谢旺盛,是体细胞胚形态建成的关键时期;POD、AMY和SOD 3种同工酶的酶谱及表达强弱在形态建成的不同时期呈现有规律的变化,可作为香果树体细胞胚发生发育特定时期的参考标记. 与胚性愈伤组织的诱导及体细胞胚的发生发育密切相关.非胚性愈伤组织和胚性愈伤组织酶谱差异明显,胚性愈伤组织中EST和AMY同工酶酶带多且活性高,非胚性愈伤组织中缺乏EST和AMY同工酶表达,AMY同工酶町作为胚性细胞分化和发育的重要标志.香果树体细胞胚形态建成过程 ,球形胚时期的AMY、POD、EST同_T酶活性最强,表明这一时期生理代谢旺盛,是体细胞胚形态建成的关键时期;POD、AMY和SOD 3种同工酶的酶谱及表达强弱在形态建成的不同时期呈现有规律的变化,可作为香果树体细胞胚发生发育特定时期的参考标记. 与胚性愈伤组织的诱导及体细胞胚的发生发育密切相关.非胚性愈伤组织和胚性愈伤组织  相似文献   

8.
苜蓿组织培养中球形胚发生时特异蛋白质和同工酶分析   总被引:10,自引:2,他引:8  
试验在苜蓿组织培养中,对球形胚形成过程中特异蛋白质表达的模式、过氧化物酶及酯酶同工酶酶谱变化进行研究,结果表明:苜蓿组织培养中从胚性愈伤组织到球形胚发育的进程中,顺序消失和出现了11种中小分子量多肽;过氧化物酶同工酶酶谱发生了显著的变化;酯酶同工酶酶谱变化不大,但其总活力对于维持体细胞胚胎发生是必须的。  相似文献   

9.
2,4-D浓度对胡萝卜体细胞胚胎发生及其同工酶的影响   总被引:1,自引:0,他引:1  
在含有不同浓度2,4-D的MS培养基上生长的胡萝卜组织,培养26天后,不含2,4-D培养基上的培养物,其酯酶同工酶谱比在含有2,4-D培养基上的培养物少一条酶带8(图3)。过氧化物酶同工酶谱中,发现前者出现的酶带1(图4)在后者的酶谱中看不到。 胡萝卜的培养组织和完整的小植物皆具有分泌过氧化物酶和酯酶的能力。总过氧化物酶活力的变化与愈伤组织、胚状体和小植物的形成有一定关系。  相似文献   

10.
红豆草(Onebrychis viciaefolia Scop.)下胚轴切段在含有1mg/IBA、1mg/1 KT 的 LS培养基上培养,两周后产生愈伤组织,通过筛选、克隆得到大量的具有胚胎发生潜力的非胚性愈伤组织,当将其转移到含1mg/1BA 的 LS 培养基上后可诱导体细胞胚胎发生。应用放射性同位素液体闪烁技术测得在胚性培养的前2天,RNA 合成速度迅速上升,随后下降,第五天后又呈缓慢上升趋势,尔后平稳。蛋白质合成速度在胚性培养的第三天达到高峰,升高很快。而 DNA 合成速度变化平缓,只是在胚性培养的第五天出现一较小的峰。胚性培养过程的 DNA、RNA 蛋白质合成速度均高于非胚性培养。  相似文献   

11.
Liu C  Xia X  Yin W  Huang L  Zhou J 《Plant cell reports》2006,25(7):621-628
A rapid and effective system of somatic embryogenesis and organogenesis from the in vitro needles of redwood (Sequoia sempervirens (D.Don.) Endl.) had been established. The influences of plant growth regulators (PGRs) and days of seedlings in vitro on adventitious bud regeneration and somatic embryogenesis were studied. The process of somatic embryo formation was also observed. The results showed that embryogenic callus was induced and proliferated on Schenk and Hildebrandt (SH) medium with BA (0.5 mg/l), KT (0.5 mg/l) and IBA (1.0 mg/l). SH medium containing BA (0.5 mg/l), KT (0.2 mg/l) and IBA (0.2 mg/l) effectively promoted adventitious bud regeneration. The highest frequency (66.3%) of direct somatic embryogenesis was obtained in the combination of BA (0.5 mg/l) and IBA (0.5 mg/l). The optimal days of seedling in vitro for adventitious bud and somatic embryogenesis were 30 days and 30–40 days, respectively. The developments of somatic embryos were similar to that of zygotic embryogenesis. The result of histocytological studies indicated that proteins were gradually accumulated in the process of somatic embryo formation and there were two peaks of starch grains accumulation that one was in the embryogenic callus and the other was in the globular embryos. These results indicated that starch and protein were closely related with the energy supply and the molecular base of somatic embryogenesis, respectively.  相似文献   

12.
Somatic embryogenesis from stem and leaf explants of Quercus robur L.   总被引:2,自引:0,他引:2  
Internodal and leaf segments from pedunculate oak (Quercus robur L.) seedlings were used as explant source to induce somatic embryogenesis. Auxin treatment influenced embryogenic response, which only occurred in explants initially cultured on media containing 4 mg/l naphthaleneacetic acid (NAA) and different benzyladenine (BA) concentrations. After 6 weeks of culture on induction medium, the explants were transferred to medium supplemented with 0.1 mg/l BA and 0.1 mg/l NAA, and 4 weeks later, they were subcultured in a growth-regulator-free medium, in which somatic embryos arose through indirect regeneration on the surface of a nodular callus. Somatic embryos were induced in explants of two out of four seedling provenances. The induction frequency ranged from 16% in leaf explants to 4% in internodal explants. Somatic embryos developed two cotyledons, which were translucent or opaque-white in appearance, but anomalous morphologies were also observed. Different embryogenic lines were established and maintained by repetitive embryogenesis in multiplication medium containing 0.1 mg/l BA plus 0.05 mg/l NAA. These results indicate that tissues from explants other than Q. robur zygotic embryos are able to produce embryogenic cultures. Received: 14 July 1998 / Revision received: 2 November 1998 / Accepted: 6 November 1998  相似文献   

13.
红江橙体胚发生及影响因素的研究   总被引:4,自引:0,他引:4       下载免费PDF全文
贺红  潘瑞炽  韩美丽  李耿光   《广西植物》1998,(4):343-346
红江橙花后4周的幼果,直径约1cm左右,这时期的胚珠最适于胚性愈伤组织的诱导。以MT为基本培养基,加上适当浓度的IAA、BA和ME可显著提高胚性愈伤组织诱导率。体胚发生,以MT+IAA0.1mg1-1+BA(ZT)1mg1-1培养基较好,能诱导产生正常体胚,且频率较高。胚性愈伤组织继代次数对体胚发生也有一定的影响,继代6次以内,胚性愈伤组织具有旺盛产生体胚能力;继代至第9次时,产生体胚的能力明显下降;至第12代时,不能产生胚状体。成熟胚转换成小植株,以MT+GA2mg1-1+NAA0.1mg1-1培养基成苗率最高。  相似文献   

14.
Plant regeneration in Arachis pintoi was obtained via two developmental pathways: organogenesis and somatic embryogenesis. Organogenic callus cultures were initiated from pieces of leaf on MS medium supplemented with NAA or 2,4-D in combination with BA, KIN or 2iP. The most suitable combination for plant regeneration through organogenesis was an initial medium composed of 10 mg/l NAA+1 mg/l BA followed by transfer of the callus to a shoot induction medium (MS+1 mg/l BA). Rooting of regenerated shoots was readily achieved by culture on MS+0.01 mg/l NAA. Embryogenic callus cultures were initiated from pieces of leaf on MS medium supplemented with PICL in combination with KIN, ZEA, BA or 2iP, and the most suitable combinations were 20 mg/l PICL+1 mg/l BA or 2iP. When pieces of embryogenic callus were subcultured on MS+1 mg/l BA, somatic embryos were differentiated and developed further into well-developed plants in MS+1 g/l AC followed by MS medium devoid of plant growth regulators. Received: 29 April 1999 / Revision received: 24 November 1999 / Accepted: 18 December 1999  相似文献   

15.
Fast growing calli induced from hypocotyl segments ofGentiana crassicaulis were used for preparation of protoplasts. High yields of viable protoplasts were produced in an enzyme solution containing 1–2% cellulase, I% pecfinase, and 0.5% Hemicellulase. Protoplasts were cultured in KM8P medium containing 1 mg/l 2,4-D, 0.5 mg/l 6BA, 500 mg/l LH, 0.5 M glucose and 0.1 M mannitol by the solid-liquid dual layer culture method. First division occurred within 4–5 days of culture at a frequency of 17.8%. Sustained divisions led to callus formation. Periodically diluting the cultures with freshly prepared liquid medium containing 1% glucose was critical for colony formation. Protocolonies about 2 mm in size were transferred onto MS medium supplemented with 3 mg/l ZT, 2 mg/l 6BA, 1 mg/l GA3, 1 mg/l NAA and 6% sucrose to obtain embryogenic calli. Plantlets were regenerated via somatic embryogenesis at high frequency on hormone-free MS Medium.Abbreviations 6BA 6-benzylaminopurine - NAA naphthaleneacetic acid - 2,4-D 2,4 - dichlorophenoxyacetic acid - ZT zeatin - GA3 gibberellic acid - LH lactalbumin hydrolysate - MES 2-(N-morpholino)-ethane sulfonic acid - MS Murashige & Skoog's medium(1962)  相似文献   

16.
GU  ZHUPING 《Annals of botany》1987,60(3):309-313
Callus of sainfoin (Onobrychis viciifolia Scop.) was initiatedfrom stem and root explants which were obtained from seedlingsgrowing in vitro, on Linsmaier Skoog (LS) medium supplementedwith 1 mg l–1 2, 4-D and 1 mg l–1 BA or only 1 mgl–1 BA, and the Vacin and Went medium without hormones.Somatic embryos were formed on LS medium containing 1 m l–1BA. Embryos developed into complete plants on filter paper saturatedwith hormone-free LS medium. Onobrychis viciifolia, somatic embryogenesis, callus culture, plant regeneration  相似文献   

17.
Regeneration of Acacia mangium through somatic embryogenesis   总被引:2,自引:0,他引:2  
 Somatic embryogenesis and whole plant regeneration were achieved in callus cultures derived from immature zygotic embryos of Acacia mangium. Embryogenic callus was induced on MS medium containing combinations of TDZ (1–2 mg/l), IAA (0.25–2 mg/l) and a mixture of amino acids. Globular embryos developed on embryogenic callus cultured on the induction medium. Nearly 42% of embryogenic cultures with globular embryos produced torpedo- and cotyledonary-stage embryos by a two-step maturation phase. The first stage occurred on 1/2-strength MS basal medium containing 30 g/l sucrose and 5 mg/l GA3 followed by the second stage on 1/2-strength MS basal medium containing 50 g/l sucrose. Of the cotyledonary-stage somatic embryos, 11% germinated into seedlings that could be successfully transferred to pots. Light- and scanning electron microscopy showed that the somatic embryos originated from single cells of the embryogenic callus. Further, a single cell layer could be detected beneath the developing somatic embryos that appeared to be a demarcation layer isolating the somatic proembryonic structure from the rest of the maternal callus. A suspensor-like structure connected the globular embryos to the demarcation layer. This is the first successful report of plant regeneration through somatic embryogenesis for this economically important tropical forest species. Received: 20 January 2000 / Revision received: 28 September 2000 / Accepted: 29 September  相似文献   

18.
An efficient somatic embryogenesis system has been established in Catharanthus roseus (L.) G. Don in which primary and secondary embryogenic calluses were developed from hypocotyls and primary cotyledonary somatic embryos (PCSEs), respectively. Two types of calluses were different in morphology and growth behaviour. Hypocotyl-derived embryogenic callus (HEC) was friable and fast-growing, while secondary callus derived from PCSE was compact and slow-growing. HEC differentiated into somatic embryos which proliferated quickly on medium supplemented with NAA (1.0 mg l−1) and BA (1.5 mg l−1). Although differentiation and proliferation of somatic embryos were faster in primary HEC, maturation and germination efficiency were better in somatic embryos developed from primary cotyledonary somatic embryo-derived secondary embryogenic callus (PCSEC). At the biochemical level, two somatic embryogenesis systems were different. Both primary and secondary/adventive somatic embryogenesis and the role of plant growth regulators in two modes of somatic embryo formation have been discussed.  相似文献   

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