抗生素对火炬松的器官发生植株再生的影响(英文)

本文研究了氨苄青霉素、羧苄青霉素和头孢霉素等3种抗生素对火炬松愈伤组织的生长和分化及不定芽生根的影响。结果表明,头孢霉素最有利于愈伤组织的诱导和生长,羧苄青霉素最有利于芽的分化,氨苄青霉素降低了不定芽的生根频率。所有试验的3种抗生素提高愈伤组织的形成和芽再生,但降低了芽的生根频率。这些结果表明,选择合适的抗生素对优化火炬松遗传转化体系有重要作用。图3表4参25。     

Influences of antibiotics on plantlet regeneration via organogenesis in Ioblolly pine （Pinus taeda L.）

Three antibiotics ampicillin, carbenicillin, and cefotaxime were evaluated for their effects on induction, growth, and differentiation of organogenic calli, as well as rooting of regenerated shoots of three Ioblolly pine (Pinus taeda L.) genotypes. Of the antibiotics administered, cefotaxime maximally increased the frequency of callus formation and growth rate of organogeni ccalli, carbenicillin maximally increased the frequency of shoot regeneration and the average number of adventitious shoots per piece of organogenic callus, ampicUlin maximally decreased the rooting frequency of regenerated shoots and mean number of roots per regenerated shoot, in comparison with antibiotic-free media. Compared with the control, ampicillin minimally increased the frequency of callus formation, cefotaxime minimally increased the frequency of shoot regeneration, and carbenicillin minimally decreased the rooting frequency of regenerated shoots in three Ioblolly pine genotypes tested. All three antibiotics increased the frequencies of callus formation and shoot regeneration, and reduced the rooting frequency ot regenerated shoots suggested that the establishment of an efficient Agrobacterium tumefaciens-mediated transformation protocol for stable integration of foreign genes into Ioblolly pine need to select a suitable antibiotic. This investigation could be useful for optimizing genetic transformation of conifers.     

Caulogenic callus induction and adventitious bud formation from embryos of long-term stored seeds ofPicea jezoensis

Caulogenic calli with a high differentiation potency were induced from mature embryos ofPicea jezoensis seeds stored over a long time, for 29 years, resulting in the active formation of adventitious buds. Embryos began to induce calli within 3 weeks of cultivating on LP medium containing 3 μM BAP and 1 μM 2,4-D. Then, the calli proliferated and transformed into caulogenic calli with bud primordia in 8 weeks. The caulogenic calli increased actively with the addition of 500 mg/l ofl-glutamine in the medium. Furthermore, caulogenic calli, induced on LP medium containingl-glutamine, resulted in the formation of adventitious buds, which elongated after transferring the calli into LP medium with 0.1 μM BAP, but withoutl-glutamine. It appears that the number of adventitious buds and the process of shoot elongation are influenced by the kind of nitrogen contained in the medium for callus induction. A part of this study was presented at the 107th Annual Meeting of the Japanese Forestry Society (1996).     

Effect ofl-phenylalanine on rooting of shoots from axillary buds of adultLarix leptolepis

Rooting of shoots derived from axillary buds was examined to establish an efficient shoot culture system of clonal micropropagation in adult tree ofLarix leptolepis Gord. (Japanese larch). Nine out of ten shoots induced calli (90%) on their shoot bases, and the two of them formed root primordia with a red pigment (20%) on the calli surface within 5 weeks after culturing on modified Murashige and Skoog (MS) medium supplemented with 1.5 μM of indolebutyric acid (IBA) and 1.5 μM of naphthaleneacetic acid (NAA). However, the primordia did not elongate actively. The addition of 10 mMl-phenylalanine in the MS medium with the auxins resulted in the formation of roots at high frequency, about 80%, and they elongated actively. Although callus was formed in all the shoots cultured on the medium withl-phenylalanine, it appeared that the callus development was less as compared to the medium withoutl-phenylalanine. Consequently, the rooting might be associated with the suppression of the induced callus.     

花烛组织培养的研究

为了找出一条适合花烛工厂化生产和最佳组织培养程序,以茎尖,叶,根等作为外植体诱志愈伤组织发生,进而诱导不定芽的发生。结果表明：黑暗有利于花烛愈伤组织的地和生长,其最佳培养基为ＭＳ＋６－ＢＡ０．７ｍｇ／Ｌ＋ＫＴ０．３ｍｇ／Ｌ。光照有利于芽的发生,其最佳培养基为ＭＳ＋６－ＢＡ０．５ｍｇ／Ｌ＋ＫＴ０．５ｍｇ／Ｌ。     

巨桉无性系EG5叶片高效再生体系的建立

以巨桉栽培无性系EG5无菌苗的叶片为外植体进行愈伤组织诱导与植株再生研究。结果表明:愈伤组织高效诱导和不定芽分化的最适培养基为改良MS+0.12 mg·L-1TDZ+0.25 mg·L-1NAA;硝酸铵对EG5叶片愈伤组织生长及植株分化的影响较大,最适宜质量浓度为0.412 5 g·L-1;最佳伸长培养基配方为改良MS+0.3 mg·L-16BA+0.05 mg·L-1IBA+0.3 mg·L-1IAA;暗培养10 d能促进不定芽分化,延缓愈伤组织老化和褐变速度。生根培养基为改良MS+0.4 mg·L-1NAA时生根率最高,为65.47%,移植成活率为90%以上。     

紫穗槐茎段愈伤组织诱导及再生体系的建立

以紫穗槐茎段为研究对象,确定了影响其愈伤组织诱导的关键因子、不定芽分化及生根培养的最佳条件,建立了紫穗槐稳定高频再生体系。试验结果表明：紫穗槐茎段愈伤组织的最适诱导培养基为MS＋6-BA4.0mg/L＋NAA2.0mg/L＋2,4-D0.5mg/L,诱导率为100%,经愈伤组织分化不定芽的最佳培养基为MS＋6-BA0.5mg/L＋NAA0.1mg/L＋KT2mg/L,分化率为96.2%,增殖倍数为7.3;不定芽最佳生根培养基为1/2MS＋酵母提取物0.5mg/L;再生植株移植在选用田园土、蛭石（5∶1）混合的基质上,光强3000lx,光照时间14h/d下生长,3周后成活率达85.67%。     

Non-aerated liquid culture promotes shoot organogenesis in <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> Labill

Eucalyptus is very recalcitrant to in vitro culture. In this research, an efficient shoot organogenesis system was developed using 60-day-old plants of Eucalyptus globulus grown in vitro and non-aerated liquid medium to improve shoot proliferation. Cultures were initiated with hypocotyls and leaf segments from plantlets cultivated on semisolid ½ MS modified medium supplemented with 4.44 µM 6-Benzyladenine (BA) and 16.1 µM 1-Naphthaleneacetic acid (NAA). Calli were transferred to shoot induction medium, with either 0.5 or 2.7 µM NAA. Shoot multiplication was carried out on 4.44 µM BA + 0.5 µM NAA medium, and semisolid and non-aerated liquid systems were compared for improving shoot proliferation. Rooting of adventitious shoots was evaluated on medium containing NAA or Indole-3-butyric acid -IBA (5 and 16 µM). Callogenesis was obtained from both types of explants, although shoot formation was only obtained from leaf-derived calli. Shoot proliferation on 4.44 µM BA + 0.5 µM NAA resulted in the most shoots/callus. Non-aerated liquid medium was more efficient in promoting shoot multiplication (53.5 shoots/callus) than was semisolid medium (28.5 shoots/callus). Levels of phenolic compounds were significantly reduced in the shoots cultivated in liquid medium. Efficient rooting (76%) was obtained using 16 µM IBA.     

In vitro plant regeneration from cotyledon-derived callus cultures of leguminous tree Gleditsia caspica Desf.

An in vitro regeneration system was developed using organogenic callus derived from in vitro grown cotyledonary explants of Gleditsia caspica Desf., an important leguminous tree. Murashige and Skoog (MS) basal medium augmented with 0.2 g L^?1 myo-inositol and various concentrations of either 2,4-dichlorophenoxyacetic acid (2,4-D), naphthaleneacetic acid, or indole-3-butyric acid (IBA) alone as well as combined with cytokinins was used for callus induction. The highest frequency of organogenic yellowish-white and nodular callus (93 %) was obtained from explants grown on medium supplemented with 13.5 μM 2,4-D and 4.4 μM benzyladenine (BA). The yellowish-white and nodular callus when transferred to MS medium supplemented with BA (2.2–17.7 μM) or kinetin (KT; 2.3–18.8 μM) solely or in combination with 2.3 μM 2,4-D produced several microshoots after 5 weeks culture. The calli cultured on MS medium with 4.4 μM BA singly showed superior growth response and produced both maximum shoot regeneration (94 %) and the highest mean number (4.3) of microshoots per callus. Transfer of regenerated microshoots onto modified MS basal medium fortified with 5.8 μM gibberellic acid and 4.4 μM BA resulted in the maximum number of internodes per shoot and the highest shoot elongation after a period of 6 weeks. Optimum rooting of 90 %, an average 6.1 roots per shoot, and a mean root length of 3.6 cm was observed when half-strength MS medium was supplemented with 9.8 μM IBA and 0.92 μM KT. The regenerated healthy plants with well-developed shoots and roots showed a survival rate of 77 % after acclimatization and transplanting to garden soil for a 10-week hardening period under ex vitro conditions.     

Establishment of an in vitro plant regeneration protocol for Casuarina cunninghamiana Miq. via indirect organogenesis

An in vitro plant regeneration protocol via indirect organogenesis from morphogenetic callus was established for Casuarina cunninghamiana Miq. Effects of plant growth regulator NAA (naphthaleneacetic acid) and BAP (6-benzylaminopurine), sucrose and AgNO₃ on callus induction, adventitious bud differentiation and shoot development were examined. Explants used were epicotyl fragments from 45-day-old seedlings. The largest callus (4.29 mm in diameter) was obtained after 1 month on a basic culture medium consisting of Murashige and Skoog ? macro- and full strength micro- elements, Nitsch and Nitsch vitamins, supplemented with 0.54 μM NAA, 3.30 μM BAP, and 30 g L⁻¹ sucrose. The calli were subcultured in the same medium above for 2 months. They were then cultured for another 2 months for adventitious bud differentiation and shoot development. The highest mean adventitious bud differentiation, number of shoots formed per callus and number of shoots ≥2 cm long per callus (47.50%, 27.38 and 4.75, respectively) were achieved on the above medium modified with NAA at 0.27 μM and supplemented with AgNO₃ 1 mg L⁻¹. Shoots were successfully rooted without plant growth regulator and the rooted plantlets survived and grew normally. This protocol for in vitro plant regeneration provides a tool not only for vegetative propagation but also for plant genetic transformation and gene function studies of C. cunninghamiana.     

中金系列杨树的组培技术研究

中金系列杨树是杨柳科杨属白杨派植物,选择其腋芽（或顶芽）、嫩茎段、嫩叶片等为外植体,进行愈伤组织诱导试验、不定芽分化试验、生根试验,筛选出了适合植株分化、生根的培养基成分;对生根试管苗进行移栽基质配比试验,确定组培苗最佳生长环境。总结出适合中金系列杨树组培的技术流程,保证了杨树的遗传稳定性,降低了培养成本,加快了培养速度。     

灯盏细辛离体叶片再生植株的研究

以灯盏细辛（Erigeron breiscapus）叶片为外植体,研究不同培养基配方对灯盏细辛离体培养过程中愈伤组织、不定芽形成,壮苗培养和离体植株生根等方面的影响。研究结果表明,低浓度的细胞分裂素与生长素配比可诱导灯盏细辛愈伤组织产生,其中MS＋BA1．00mg／L＋NAA0．50mg／L培养基配方,愈伤组织诱导率达到99．22％;不定芽诱导需要较高浓度的细胞分裂素,适宜培养基MS＋KT4．00mg／L＋IBA0．50mg／L的不定芽诱导率为38．51％,诱导系数为0．49;MS＋BA0．50mg／L＋NAA0．50mg／L＋水解酪蛋白1000．00mg／L＋PVP1000．00mg／L＋GA0．50mg／L培养基可促进不定芽分化生长,形成离体植株;离体植株生根需要高浓度生长素,适宜培养基为1／2MS＋BA0．50mg／L＋NAA3．00mg／L＋IBA3．00mg／L＋活性炭0．30％。     

野葛愈伤组织诱导与不定芽分化

采用MS为基本培养基，附加NAA、6-BA和IAA3种激素．诱导野葛不同外植体愈伤组织形成；再将愈伤组织接种在不同浓度6-BA的MS培养基上．分别在光照和黑暗条件下进行不定芽的诱导。结果表明．野葛幼叶、茎段和顶芽在适宜激素的诱导下均能形成愈伤组织，但不同激素组合其愈伤组织诱导率不同。幼叶愈伤组织诱导的最适培养基为MS NAA1．0mg／L 6-BA1．0mg/L．其诱导率为75％．茎段愈伤组织诱导的最适培养基为MS NAA1．0mg／L 6-BA3．0mg／L IAA0．2mg/L，其诱导率为70％。不同外植体形成的愈伤组织其不定芽的分化诱导率不同．茎段愈伤组织在不同浓度6-BA下均能诱导出不定芽；光照有利于芽的分化．在光培养条件下，茎段愈伤组织不定芽平均诱导率为66．7％。     

沟叶结缕草的组织培养和无性系的建立

通过研究不同基本培养基及植物生长调节组合对沟叶结缕草丛生芽诱导、增殖和愈伤组织诱导、分化的影响,建立起沟叶结缕草试管无性系。以地下匍匐根状茎的顶芽为外植株,在MS 6-BA 2 mg.L-1 NAA 0.1 mg.L-1培养基上诱导丛生芽形成,以MS KT 3 mg.L-1 NAA 0.1 mg.L-1作为继代增殖培养基,建立起丛生芽苗→不定芽发生→丛生芽苗速生试管无性系;以丛生芽苗基部为外植体,以MS 2.4-D 4 mg.L-1作为愈伤组织诱导培养基,以MS基本培养基作为愈伤组织分化培养基,建立起丛生苗→愈伤组织→愈伤组织分化→丛生苗试管无性系。试管无性系室外移栽成活率达95%以上。沟叶结缕草试管无性系的建立为细胞工程育种和基因转化的研究奠定了基础。     

花叶菖蒲组织培养与快繁技术试验

取花叶菖蒲的茎尖作外植体，培养于附加不同激索配比的MS培养基匕进行不定芽诱导、增殖及试管苗生根等试验，结果表明，6种配比培养基中，最适合花叶菖蒲不定芽诱导的培养基为MS＋6-BA5mg／L；增殖培养基为MS＋6-BA5Mg／L＋NAA0．5mg／L；试管苗生根诱导前在空白Ms培养基先壮苗培养一代，生根培养基为MS＋NAA0．5mg／L。试管苗不经炼苗可直接出瓶，移栽于蛭石：珍珠岩为1：1的混合基质中，成活率可达98％以上。     

柔枝松组培苗不定根形成的影响因子探讨

试验以组织培养获得的柔枝松不定芽为对象,对其进行不定根诱导研究。结果发现：IBA对不定根的诱导无明显影响。IBA和NAA配合使用时,形成了大量畸形不定根。NAA对不定芽的伸长生长有明显的促进作用,当培养基中添加0．05mg·L^-1NAA时,苗高达3．12cm,显著高于其他处理。不同浓度NAA及其不同诱导时间对不定根的形成无明显促进作用,反而促进了大量愈伤组织的形成。总之,柔枝松不定芽在无植物生长调节剂的1／2GD培养基能稳定诱导不定根形成,最高生根率为16．67％,说明柔枝松不定芽可能易诱导潜伏根原基,并在无植物生长调节剂的刺激的培养条件下伸长生长。IBA和NAA的使用浓度和诱导时间仍需进一步研究。     

In vitro regeneration of plantlets from mature embryos of Pinus ayacahuite

A plantlet regeneration protocol was developed for Pinus ayacahuite var. ayacahuite (Ehrenb.). Embryos from mature seeds from ten provenances were cultured in a 16-h photoperiod for 3 days on a medium containing 30 mM sucrose and 0.7% agar. Cotyledons from these embryos were subcultured onto MCM medium (Bornman 1983) supplemented with 50 micro M N(6)-benzyladenine and 90 mM sucrose for 2 weeks. Bud development and shoot elongation were maximized by subculturing the explants on half strength AE medium (von Arnold and Ericksson 1981), supplemented with 60 mM sucrose and 0.05% activated charcoal every 30 days. Seed source had a significant effect on the responses of the embryos to the bud induction protocol. For the provenance with the best response to bud induction, about 79% of the cultured cotyledons formed buds, and each cotyledon formed a mean of 9.1 buds, so that about 70 shoots could be induced from each seed. The best rooting response (40% rooting) was obtained by treating the shoots for 8 h with 100 micro M naphthalene acetic acid.     

红千层愈伤组织诱导及植株再生

以茎段、芽和叶片为材料,探讨了红千层愈伤组织诱导及植株再生的方法.结果表明:红千层的茎段、芽和叶片均可诱导出愈伤组织,通过继代培养可发育成绿色和粉红色2种类型的愈伤组织,其中绿色、致密的愈伤组织可以分化出不定芽;培养基1/2M S 6-BA 1.0 m g/L NAA 0.1 m g/L适宜愈伤组织不定芽的诱导,在培养基1/2M S IBA 0.25 m g/L上试管苗的生根率可达95%.     

红千层愈伤组织诱导及植株再生

以茎段、芽和叶片为材料,研究了红千层愈伤组织诱导及植株再生的方法。结果表明:红千层的茎段、芽和叶片均可诱导出愈伤组织,通过继代培养愈伤组织可发育成绿色和粉红色2种类型,其中绿色、致密的愈伤组织可以分化出不定芽。培养基1/2M S 6-BA 1.0 m g/L NAA 0.1 m g/L适宜愈伤组织不定芽的诱导,在培养基1/2M S IBA 0.25 m g/L上试管苗的生根率可达95%。