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1.
Carimi Francesco Tortorici Maria Concetta De Pasquale Fabio Crescimanno Francesco Giulio 《Plant Cell, Tissue and Organ Culture》1998,54(3):183-189
Somatic embryogenesis was induced and plant regeneration was obtained in 11 different genotypes of sweet orange navel group
[Citrus sinensis (L.) Osb.] from cultures of stigma/style explants and undeveloped ovules. Explants were cultured on 3 different
modifications of Murashige and Skoog medium: 500 mg l-1 malt extract; 500 mg l-1 malt extract and 4.6 μM kinetin; and 500
mg l-1 malt extract and 13.3 μM 6-benzylaminopurine. Sucrose (146 mM) was used as carbon source. Somatic embryogenesis occurred
1–3 months after culture initiation from undeveloped ovule and stigma/style cultures of all the genotypes tested. Somatic
embryos developed into plantlets with a high frequency (74%) after transfer to Murashige and Skoog medium supplemented with
146 mM sucrose and 500 mg l-1 malt extract. Plants were successfully transferred to soil.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
2.
Zygote implantation to cultured ovules leads to direct embryogenesis and plant regeneration of wheat 总被引:2,自引:0,他引:2
Jochen Kumlehn Reinhold Brettschneider Horst Lörz Erhard Kranz 《The Plant journal : for cell and molecular biology》1997,12(6):1473-1479
Direct embryogenesis and plant regeneration were obtained by implantation of individual wheat ( Triticum aestivum L.) zygotes into cultured ovules of wheat or barley. The zygotes were isolated mechanically from emasculated spikes, 3–9 h after hand-pollination. In 13 independent experiments, a total of 186 zygotes were implanted into excised ovules obtained from emasculated spikes which had been treated previously with 2,4-dichlorophenoxyacetic acid to induce parthenocarpic, embryoless ovary development. On average, 17.2% of the implanted zygotes gave rise to dorsiventrally differentiated embryos. The embryos resembled those growing in planta with no obvious deviation from the zygotic embryogenesis pathway. In contrast to previously described regeneration systems from individual zygotes of higher plants, this is the first study in which direct embryo formation is reproducibly obtained without intermediate tissue dedifferentiation. Most embryos germinated when transferred to regeneration medium, and later formed phenotypically normal, fully fertile plants. Regenerants were confirmed to be derived from the implanted zygotes by means of AFLP and/or morphological analyses. Although zygote implantation has long been established as a useful method in sexual animal reproduction, an equivalent technique for plants is described here for the first time. Since the zygotes enter the embryogenic pathway directly, the genome is presumably as stable as during embryogenesis in planta . With this new approach, isolated wheat zygotes are accessible to micromanipulation without affecting their subsequent embryonic development. 相似文献
3.
Azahara Barra-Jiménez Miquel Blasco Mar Ruiz-Galea Cristina Celestino Jesús Alegre Isabel Arrillaga Mariano Toribio 《Trees - Structure and Function》2014,28(3):657-667
Key message
Integuments from holm oak developing ovules were suitable initial explants to obtain embryogenic lines from which plants could be regenerated.Abstract
The implementation of multivarietal forestry as part of breeding strategies is expected to provide more productive forest plantations. To achieve this, a reliable and effective method for mass production of clonal plants is needed. Somatic embryogenesis is considered the enabling technology for implementing multivarietal forestry. The holm oak (Quercus ilex L.) is a Mediterranean evergreen tree of economic interests because of the acorn production for animal feed and edible fungi mycorrhization. The aim of this work was to obtain clonal plants by inducing somatic embryogenesis in tissues of female flowers from mature trees. The influence of the developmental stage of the explant, the genotype and medium composition, and the effect of arabinogalactan proteins on the induction frequency, were assessed. Somatic embryogenesis induction (frequency ranging from 1.2 to 3.2 %) was restricted to ovules excised at an advanced stage of development, when they were at least 3–4 mm wide and the rest of the ovules within the ovary had aborted. Somatic embryos arose from the integuments of those fertilized ovules. Embryogenic response was obtained on media with and without plant growth regulators. All the genotypes that were cultured on medium containing “as reported by Schenk and Hildebrandt (Can J Bot 50:199–204, 1972)” SH macronutrients could be captured. Treatments including Larix arabinogalactan proteins did not improve induction, while those from Acacia inhibited the embryogenic response. Several embryogenic lines were multiplied by repetitive embryogenesis on medium lacking plant growth regulators. Mature somatic embryos of three genotypes were germinated at frequencies ranging from 41 to 58 %, and converted into plants at frequencies from 11 to 30 %, depending on the genotype. 相似文献4.
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
5.
Explants excised from the young shoots of Aralia elata (Miq.) Seem. were cultured on MS media. Calli were induced from the explants on MS medium supplemented with 0.5 mg/L 2, 4-D, 0.5 mg/L BA and 0.5 mg/L NAA. Then these calli were transferred onto the MS medium containing 2.0 mg/L 2,4-D + 0.5 mg/L BA + 0.5 mg/L NAA and 0.2% activated charcoal. Under these conditions the somatic embryoids were observed and regenerated plants were obtained from somatic embryogenesis. Then, a experimental system with stability and high regenerating efficiency has been set up for the propagation of the young plants, the cell breeding technology and the control of somatic embryogenesis of Aralia elata (Miq.). 相似文献
6.
Efficient plant regeneration via somatic embryogenesis has been developed in pigeonpea. Cotyledon and leaf explants from
10-day-old seedlings produced embryogenic callus and somatic embryos when cultured on Murashige and Skoog (MS) medium supplemented
with 10 μm thidiazuron (TDZ). Subsequent withdrawal of TDZ from the induction medium resulted in the maturation and growth of the embryos
into plantlets on MS basal medium. The rooted plantlets were transferred and acclimatized on vermiculite where they showed
normal morphological characters.
Received: 23 December 1996 / Revision received: 22 July 1997 / Accepted: 2 August 1997 相似文献
7.
C. Arias-Castro A. H. Scragg A. Stafford M. Rodriguez-Mendiola 《Plant Cell, Tissue and Organ Culture》1993,32(1):77-82
Sweet potato (Ipomoea batatas (L.) Lam.) breeding has been hampered by self-and cross-incompatibilities that are frequently encountered among the plants in the section Batatas. Ovule culture techniques were developed to assist in overcoming some of these incompatibilities. Ovules that contain embryos at the late globular to heart shaped stage of development were cultured on MS medium containing full strength or one-half strength salts with 3%, 8% or 12% sucrose. Ovules were cultured either intact or after slicing. Ovules of I. triloba and I. trifida were successfully cultured as early as 3 and 4 days after pollination while sweet potato ovules were successfully cultured 5 and 6 days after pollination. The percentage of ovules with developing embryos on the media tested ranged from 27.8% to 50.2%. The highest percentage of embryos developed when the ovules were sliced and cultured on medium containing one-half MS salts and 8% sucrose. Three plants were recovered from cultured ovules of incompatible interspecific crosses.Abbreviations DAP
days after pollination
- MS medium
Murashige and Skoog (1962) medium 相似文献
8.
Partial endosperm development without paternal genome involvement was induced in unpollinated ovaries of wild-type Arabidopsis thaliana cultured in vitro. Unpollinated pistils were cultured on hormone-free Murashige and Skoog (MS) medium with addition of 6% sucrose and supplemented with: benzylaminopurine (BAP; 2 mg l–1) combined with naphthylacetic acid (NAA; 0.1 mg l–1), 2,4-dichlorophenoxyacetic acid (2,4-D; explants exposed to 1-h auxin shock 20 or 40 mg l–1, and transferred to hormone-free MS medium). Initiation of autonomous endosperm (AE) development was induced on all media used in 54 ovules from 39 cultured ovaries (26%), with an average frequency of 1.4 ovules/ovary. The highest frequency of partial endosperm formation occurred on media combining the two growth regulators BAP and NAA (59% of ovaries had ovules with AE), although endosperm development was also induced on hormone-free medium (in 20.5% of ovaries). The number of AE nuclei ranged from 2 to ~50, depending on the day of culture and medium; neither cellularization nor differentiation on specific regions typical for endosperm of wild-type Arabidopsis, were noted. Fertilization independent endosperm most probably originated from the secondary nucleus, but involvement of the polar nuclei could not be excluded, as indicated by nuclear size and structure. In vitro conditions did not influence egg cell proliferation. Gynogenic embryos were observed neither in the ovules with autonomous endosperm nuclei nor in ovules without endosperm induction. 相似文献
9.
以盐肤木(Rhus chinensis Mill.)幼胚为外植体,研究不同植物生长调节剂组合对其愈伤组织诱导及体细胞胚胎发生的影响,以建立盐肤木体细胞胚胎发生及植株再生体系。结果表明,最适愈伤组织诱导培养基为MS+6-BA 0.2 mg/L+2,4-D 1.0 mg/L,诱导率为84.57%,诱导出的初代愈伤组织白色或淡黄色,质地疏松,表面光滑,为非胚性愈伤。初代愈伤组织转移到1/2 MS+6-BA 2 mg/L+NAA 0.5 mg/L培养基上培养1个月后,长出淡黄色质地紧密的胚性愈伤组织,诱导率高达100%,在此培养基上胚性愈伤组织增殖倍数为854.73%。所获得的胚性愈伤组织转接到1/2 MS+6-BA 2 mg/L+NAA 0.5 mg/L+蔗糖4%的培养基上培养1个月后可诱导体细胞胚胎发生,诱导率可达32.67%。诱导得到的体细胞胚胎经历球形胚、心形胚、鱼雷胚、子叶胚进一步分化发育成苗。无菌苗炼苗后栽种到泥炭土∶蛭石∶珍珠岩为2∶1∶1的生长基质上,能100%稳定成活。经过细胞学观察分析,体细胞胚的发育与合子胚相似。 相似文献
10.
Somatic embryogenesis of European chestnut (Castanea sativa Mill.) was obtained using juvenile tissue cultured on P24 medium with 5 M 2,4-dichlorophenoxyacetic acid plus 0.5 M 6-benzylaminopurine (BA) for three weeks and then cultured on 0.89 M BA. Induction frequency with ovaries ranged from 2.0 to 19.1 % and was observed in tissue collected 2 to 8 weeks postanthesis, ovules used as a starting tissue gained 0.8 to 7.8 %, 3 to 9 weeks postanthesis. Zygotic embryos collected 5 to 10 weeks postanthesis formed 10.5 to 57.1 % somatic embryos, respectively. The culture lines were maintained via secondary embryogenesis on P24 medium with 0.89 M BA. Development and maturation were stimulated on P24 medium with increased agar concentration (1.1 %). Five plantlets were transferred to substrate and acclimatized successfully in greenhouse. 相似文献
11.
无核白葡萄胚挽救育种技术研究 总被引:19,自引:3,他引:16
无核白葡萄在授粉后20-50d,将胚珠接种在含不同激素的Nitsch培养基上,培养80-90d后转入胚荫发培养基中,待胚萌发20d左右,再转入成苗培养基中使其发育成正常幼苗。结果表明,胚萌发适宜的培养基是附加0.5mg/L IAA,1.5mg/L BA和0.5mg/LGA3的Nitsh培养基,在所试10个接种时期中,无授粉后39d胚的发育率最高。 相似文献
12.
本文报告了黑穗醋栗(Ribes nigrum L.)三个栽培品种中的薄皮黑豆未受精胚珠(花粉发育到单核晚期)在MS基本培养基附加植物激素BA,2.4-D和GA_3中形成了体细胞胚状体。长度约为0.5—1.0cm大小的胚状体在生根培养基中可以形成完整的再生植株。经过筛选得到了体细胞胚性愈伤组织无性系,继代培养二年多仍能保持胚状体形成能力。试验结果表明:诱导体细胞胚胎发生受品种和接种时期的影响,适宜的接种时期为花粉发育到单核晚期。培养基中的BA为诱导胚胎发生和保持胚性愈伤组织无性系所必需。 相似文献
13.
14.
Production and cryoconservation of embryogenic cultures of mandarin and mandarin hybrids 总被引:1,自引:0,他引:1
Assays were performed to obtain embryogenic callus lines from nine mandarin and mandarin hybrid cultivars by in vitro culture
of ovules collected from immature fruits six weeks after anthesis. All cultivars produced embryos and loose friable callus.
The small proliferations of nucellar callus from cultured ovules were suitable to recover embryogenic callus lines by periodical
subculturing to fresh medium. The embryogenic callus cultures were subjected to cryoprotection with 10% (v/v) (DMSO), freezing
by slow cooling, storage in liquid nitrogen and thawing by fast warming. Whole plants from these cryopreserved cultures were
recovered through embryogenesis.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
15.
Plant regeneration from protoplasts isolated from friable embryogenic callus of cassava 总被引:2,自引:0,他引:2
E. Sofiari C. J. J. M. Raemakers J. E. M. Bergervoet E. Jacobsen R. G. F. Visser 《Plant cell reports》1998,18(1-2):159-165
Protoplasts were isolated from friable embryogenic callus (FEC) and from suspensions derived from FEC of cassava genotype
TMS60444. Suspensions yielded the highest number of protoplasts (1.5×106 protoplasts/g fresh weight). Protoplasts plated at a density of 105–106/ml in a medium supplemented with 0.5 mg/l α-naphthaleneacetic acid and 1 mg/l zeatin began dividing after 3 days, and after 30 days this resulted in an absolute plating
efficiency as high as 2.5%. After 2 months of culture, 60% of the developed calli were highly friable and in appearance identical
to the original FEC. The protoplast derived FEC was first purified through two rounds of selection of 3 weeks each before
beeing cultured for regeneration of plants. This was done by culturing the protoplast-derived FEC for 11 weeks on maturation
medium, yielding a maximum of 184 organized embryos per 10.000 initially cultured protoplasts. Most of the organized embryos
were torpedo shaped and matured after they had been isolated from the calli and transferred to fresh medium. Mature embryos
were multiplied by secondary somatic embryogenesis at high efficiency (>90%) on a medium supplemented with 8 mg/l 2,4-dichlorophenoxyacetic
acid. About 30% of the mature secondary somatic embryos developed into shoots after transfer to a medium supplemented with
1 mg/l N6-benzylaminopurine (BAP). Shoots rooted readily on a medium without BAP.
Received: 30 August 1996 / Revision received: 9 June 1997 / Accepted: 1 October 1997 相似文献
16.
龙牙楤木[Aralia elate(Miq.)Seem]是五加科楤木属的多年生乔本药用植物,又称刺老鸦,具有补气、活血、祛风、利湿、止痛等功效。主要分布于我国东北地区、朝鲜、日本和俄罗斯的西伯利亚地区。其幼嫩茎叶是有名的山菜,在我国及东南亚一些国家很受欢迎。但是,近些年来由于人为恶性采伐,使 相似文献
17.
油松体细胞无性系的建立 总被引:15,自引:0,他引:15
以油松(Pinustabulaeformis)种胚为外植体,通过器官发生途径建立体细胞无性系。以MS为基本培养基,在芽的诱导和增殖培养基中,附加植物激素,以6BA+KT和NAA效果最好,两者的配比为5-10:1,6BA和KT的浓度分别均不超过1mg/L。同时根据培养物的发育状态交替使用活性炭,则对芽的增殖有明显的促进作用。选择发育状态较幼嫩的刚抽茎的外植体,诱导生根,在1/2MS+KT0.1+IBA0.1+NAA0.1mg/L的培养基中,生根率达32.%。体细胞胚胎发生途径建立细胞无性系也已取得成功,筛选出可继代培养的胚性胚柄团无性系,在胚轴和子叶上诱导出成熟的体细胞胚并获得完整小植株。 相似文献
18.
High-frequency embryogenesis and plantlet regeneration from isolated microspores of indica rice 总被引:8,自引:0,他引:8
We report high-frequency embryogenesis and plantlet development from microspores isolated from anthers of two indica (IR-43,
IR-54) and a japonica (T-309) rice cultivars, without prior nutrient preculture of anthers. Pretreatment stress of anthers
with mannitol or a sugar-starvation medium, and use of maltose as the carbohydrate source in the microspore culture medium
were found to be critical. Co-culture of microspores with rice ovaries was found beneficial but not essential. More than 60%
of the microspores of the japonica variety Taipai-309 and 25–45% of the indica cultivars IR-54 and IR-43 showed induction
of non-gametophytic development. Consequently, in the best treatments for IR-43 and T-309, more than 500 microspore-derived
embryos could be obtained from a single dish (35 mm) containing about 80,000 microspores. Among the indica cultivars, the
maximum response was obtained in the basal medium M-019. Plantlet regeneration occurred in about 9% (T-309), 7% (IR-43) and
2% (IR-54) of the transferred embryo-like structures.
Received: 6 November 1996 / Revision received: 18 June 1997 / Accepted: 20 August 1997 相似文献
19.
百合未授粉子房离体培养胚胎形成及植株再生 总被引:2,自引:0,他引:2
未受精子房离体培养是诱导雌核产生单倍体的技术之一。以1个野生种和3个杂种系共7个百合(Lilium)基因型为实验材料, 探讨了基础培养基、花蕾取样时期和外源激素等因素对百合未授粉子房离体培养胚胎形成的影响。结果表明, CBM、MS和BDS三种基础培养基均可诱导百合未授粉子房胚胎形成, 但以BDS培养基诱导效果最佳; 开花前1天的花蕾较适于百合未授粉子房离体培养; 2 mg·L-1 2,4-D + 2 mg·L-1 6-BA和2 mg·L-1 2,4-D + 2 mg·L-1 KT两种外源激素配方均适用于百合未授粉子房离体培养诱导胚胎形成。在培养过程中, 大多数胚性胚珠中只含有1个胚胎, 位于珠孔端、合子端或极核处, 少数胚性胚珠中含有双胚胎。通过百合未授粉子房离体培养, 从5个基因型材料中共获得146株再生植株。采用根尖染色体计数法对其中的62株进行了倍性测定, 其中43株与母体植株染色体倍性不同。 相似文献
20.
Susan H. Woods Gregory C. Phillips John E. Woods Glenn B. Collins 《Plant cell reports》1992,11(5-6):257-261
An efficient protocol has been developed for the in vitro propagation of Mexican Weeping Bamboo through somatic embryogenesis from zygotic embryo explants. Mature seeds and excised embryos were cultured in the light or in the dark on both Murashige and Skoog's and Gamborg's B5 basal media with various supplements. Optimal somatic embryogenesis and plant regeneration were obtained by culture in the dark on Murashige and Skoog's basal medium supplemented with 3 mg/1 2,4-dichlorophenoxyacetic acid, 0.5 mg/1 6-benzylaminopurine and 2.0% sucrose. More than 95% of the germinating somatic embryos developed shoots and roots, and were transferred to soil with 85% success. 相似文献