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1.
The effects of glucose concentrations, different sugars and combinations of 2,4-D and kinetin on cell division and colony formation were examined in cultures of protoplasts isolated enzymatically from suspension cultures of Iris hollandica N6 medium supplemented with 1 mg/l 2,4-D, 1 mg/l kinetin, 200 mg/l casein hydrolysate, 250 mg/l proline, 0.3– 0.5 M glucose and 20 g/l agarose was suitable for cell division and colony formation. When colonies formed were transferred to hormone-free MS medium, many shoots were induced. In addition, when induced shoots were transferred to MS medium with 1 mg/l NAA, root induction was observed. A plant regeneration system from protoplasts of I. hollandica was thus established. 相似文献
2.
P. B. Kirti 《Plant Breeding》1988,100(3):222-224
By using a system of agirose plating and agarose bead culture, it was possible to induce efficient somatic embryogenesis in protoplast-derived calli of two rapeseed varieties, ‘Ceres’ and ‘Duplo’. Protoplasts were isolated from hypocotyls. For the initial protoplast culture a modified 8P medium was employed containing 2,4D (1.0 mg/l), NAA (0.1 mg/ 1), BAP (0.4 mg/l) and mannitol (7 %). After microcalli were obtained in four weeks, somatic embryos were induced by a two-step method. This involved a modified MS medium containing 2,4D (3.0 mg/l) in the first step and no 2,4D, but BAP (3.0 mg/l) and GA3 (0.1 mg/l) in the second. This procedure also secured plant regeneration. 相似文献
3.
Plant regeneration from protoplasts of Iris germanica L. 总被引：1，自引：0，他引：1
Summary Protoplasts were isolated enzymatically from suspension cultures derived from embryogenic calli induced by leaf base culture of Iris germanica. In protoplast culture, the effects of glucose concentration, different sugars and combinations of 2,4-D and KIN on protoplast division and colony formation were examined. N6 medium supplemented with 0.1–1 mg/l 2,4-D, 1 mg/l KIN, 200mg/l casein hydrolysate, 250 mg/l proline, 0.2 M glucose and 20 g/l agarose was suitable for protoplast division and colony formation. When colonies formed were transferred onto hormone-free MS medium, many plantlets were regenerated through somatic embryogenesis. Thus, we could establish a plant regeneration system from protoplasts of I. germanica.Contribution from the Laboratory of Plant Breeding, Faculty of Agriculture, Miyazaki University, Japan, No. 95. 相似文献
4.
Optimization of mature embryo-based high frequency callus induction and plant regeneration from elite wheat cultivars grown in China 总被引：2，自引：0，他引：2
We have developed an efficient procedure for plant regeneration of elite wheat cultivars using mature embryos. Firstly, we established the optimal combination of basal media, inoculation method and pretreatment method using biostatistical methods. The results indicated that the combination of MS medium and longitudinally bisected mature embryos showed the highest culture efficiency, whereas the pretreatment method had no significant effects on callus induction or plant regeneration. A 70% primary callus induction rate was achieved on MS medium containing 2 mg/l 2,4‐d for all tested cultivars. Primary calli were then transferred onto the subculture medium to initiate embryogenic calli. Supplementation of the subculture medium with the appropriate combination of phytohormones (2.0 mg/l 2,4‐d , 0.5 mg/l BA and 0.1 mg/l NAA) significantly enhanced embryogenic callus production. The addition of AgNO3 (10 mg/l) in regeneration medium promoted plant regeneration, whereas CuSO4 stimulated root formation. The use of this protocol achieved successful plant regeneration in eight tested cultivars. The culture efficiency ranged from 15.3% to 36.8%, suggesting this regeneration system may be an effective alternative for wheat genetic transformation. 相似文献
5.
Development of a technique for in vitro unpollinated ovary culture in rice, Oryza sativa L. 总被引：2，自引：0，他引：2
A simple and efficient technique for in vitro unpollinated ovary culture in rice which is also applicable for indica genotypes
was developed for breeding and genetic studies. Sampling explants at the auricle distance of 7–12 cm between the two uppermost
leaves of a tiller, providing a chilling pretreatment and ovaries with 1/3 of the hulls intact gave optimum response to culture.
For callus induction with the spontaneous breaking of ovaries, N6 media supplemented with NAA (2 mg/l) and DMSO (0.6–0.8%) gave a mean PCI value of 3.8% and range of 0.8–12.5% among genotypes.
Media combining 2,4,5-T or 2,4-D with NAA in N6 medium also has reasonably good callus induction. For calli induced inside, 2,4-D (0.2–0.5 mg/l), NAA (2 mg/l) and KT (1
mg/l) contained media were superior. The maximum green plant regeneration (PPR) of 77.3% was found with the medium containing
NAA 0.25 mg/l, IAA 0.5 mg/l and KT 2.0 mg/l. Significant genotype, medium and their interaction effects for per cent ovary
survival and callus induction were observed.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
6.
Summary Callus growth and its subsequent regeneration into complete plantlets was achieved from in vitro cultured anthers ofBrassica nigra (L.)Koch. Callus was induced on a modified N6 medium containing trace elements, organics of B5 medium and 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). Morphogenesis of callus in the form of shoots on MS medium containing indole-3-acetic acid (IAA) and N6-benzyladenine (BA) 0.5 mg/l each and embryoids on MS medium containing 0.5–1.0 mg/l IAA and 3.0–5.0 mg/l BA could be accomplished. Chromosomal analysis revealed presence of 41% haploids (n=8) amongst the regenerated plants. 相似文献
7.
In order to develop fully inbred chicory plants, dihaploid plants were raised from callus derived from microspores of three selected Witloof, Robin and Treviso types. Microspores were isolated from florets containing pollen at the uninuclear state and cultured in a modified MS medium plus 0.5mg/l 2,4-D, 0.5mg/l IAA and 2.0mg/l zeatin. During culture periods of up to 6 months, gametoplasts emerged from pollen grains, divided and started to form colonies and calli. These were subcultured on the same basal medium supplemented with 0.5mg/l BA and 0.5mg/l IAA. Shoot growth was enhanced on a low salt-containing medium supplemented with 0.4mg/l kinetin and 0.2mg/l IAA. Shoots were rooted on a half-strength Lepoivre medium plus 0.2mg/l IBA and finally transferred to soil. Florets were excised from 34 capitula, but only microspores from four of them developed into plants via callus. More than 450 plants were raised in the greenhouse and the field. Leaves from these plants were subjected to DNA fluorescence analysis via flow cytometry: a range of ploidy levels was detected. The cell composition of 44 of these plants was predominantly haploid, with a diploid background. Regenerant plant phenotypes were compared with the parent genotypes. The value of such haploids in commercial chicory breeding is discussed. 相似文献
8.
大豆幼荚子叶原生质体培养及植株再生 总被引：9，自引：0，他引：9
本文研究了13个栽培大豆（Glycine max L.）品种原生质体培养的再生能力。从大豆幼荚子叶酶解游离原生质体,用Gellan Gum进行株状包埋,悬浮在含2,4-D 0.1-0.2mg/L,BA0.5-1.0mg/L的改良MS液体培养基中,原生质体培养3天后开始第一次分裂,以后持续分裂。供试基因型间的10天植板率差异显著,变幅为33-67%。30天内形成大量的 相似文献
9.
Copper tolerance of Nicotiana tabacum L. var. Xanthi in vitro was achieved through plant regeneration from leaf explants on Murashige and Skoog's (MS) medium supplemented with 0.5 mg/l BA, 0.1–0.25 mg/l IAA and 60 μ m Cu. Tolerant organogenic calli showed more vigorous growth in medium containing 60 μ m Cu than the non-tolerant calli. Standard growth parameters such as fresh and dry weight of organogenic callus, growth tolerance index (GTI), enzyme activity (peroxidase and catalase) and copper accumulation were used as indicators of copper tolerance. The activities of peroxidase and catalase as well as estimation of protein, total amino acid and chlorophyll were greater in tolerant calli than non-tolerant ones. The GTI in the 4 weeks after the beginning of treatments yielded significant differences among the tolerant and non-tolerant organogenic callus cultures. The accumulation of copper in the tolerant calli increased significantly with an increase in copper concentration in the medium. Shoot bud regeneration was achieved in both tolerant and non-tolerant organogenic calli on MS medium containing 0.5 mg/l BA and 0.1 mg/l IAA. The tolerant regenerated shoots were rooted on half-strength basal MS medium with 60 μ m Cu for selection of tolerant clones. This study may help in the selection and characterization of Cu-tolerant lines of N. tabacum cv. 'Xanthi' for building conservation strategies and also for phytoremediation programmes. 相似文献
10.
Protocols of plant regeneration have been developed for Brassica carinata for creating somaclonal variation for plant type and adaptability, so that this species can fit into cropping systems in Indian agriculture. The response of cotyledonary and stem explants was assessed for callus induction and shoot regeneration on MS and B5 basal media containing different combinations of auxin and cytokinin concentrations. MS medium supplemented with BA and NAA favoured callus induction. Supplementing MS with combinations of BA and IAA, as also with BA alone, regenerated shoots from the ex pi ants with a high frequency. The frequency of shoot regeneration and the mean number of shoots per explant were higher in cotyledons than in stem explants on identical growth regulator combinations. On B5 medium, supplemented with BA (2 mg/l) and IBA (0.4 mg/l), compact callus was produced which regenerated shoots on transfer to medium containing BA (0.8 mg/l). Genotypic differences among carinata accessions for regeneration were also observed. 相似文献
11.
Mohsen Hesami Mohammad Hosein Daneshvar 《Journal of Crop Science and Biotechnology》2018,21(2):129-136
The aim of this study is to introduce the suitable protocol for indirect regeneration from seedling-derived leaf segment of Ficus religiosa. The leaf explant successfully produced callus on MS medium containing various concentrations of auxin in combination with BAP. The maximum callus induction (100%) was achieved in MS medium containing 0.5 mg/l 2,4-D plus 0.05 mg/l BAP and MS medium containing 1.5 mg/l NAA plus 0.15 mg/l BAP as well. MS medium consisting of 2,4-D produced yellow-brownish and friable callus (type I) while the yellowish and compact calli (type II) were obtained in MS medium consisting of NAA. On the other hand, MS medium supplemented with IBA formed greenish and compact calli (type Ш). The regeneration rate in type II callus was less than the type I, and there was no shoot induction observed on type Ш calli. MS medium supplemented with 1.5 mg/l BAP in combination with 0.15 mg/l IBA had the highest regeneration frequency (100%) and maximum shoot numbers (5.16) as well as shoot length (2.56 cm) in type I callus. A maximum of 93.33% root induction was observed in MS medium supplemented with 2.0 mg/l IBA plus 0.1mg/l NAA. The plantlets were successfully transferred to the greenhouse. This system could be utilized for large-scale multiplication of Ficus religiosa. 相似文献
12.
A protocol for high frequency callus induction and plant regeneration from sunflower (Helianthus annuus L.) anthers is described.
Different variables using Murashige & Skoog (MS) basal medium supplemented with 2.0 mg/l α-naphthaleneacetic acid (NAA) and
1.0 mg/l N6-benzyladenine (BA) were tested for their ability to enhance the frequency of anther callusing and subsequent embryogenesis.
Of these, agar concentration, sucrose concentration, carbohydrate source had significant effect on callusing, while differences
due to incubation under dark vs light conditions, cold pretreatment of capitula for 1 to 6 days prior to anther inoculation
and genotype on callusing were non-significant. However, all these factors exerted highly significant influence on embryogenesis
when calli from the various media were transferred to medium supplemented with 0.1 mg/l NAA and 0.5 mg/l BA. With the procedure
developed, callusing as high as 100% and embryo formation at a frequency of 44% was achieved. Although complete embryos were
formed the frequency of their conversion to whole plantlets was low (14.3%). Hence, the embryogenic pathway was bypassed to
obtain multiple shoots by transferring embryogenic calli with developing embryos to MS medium supplemented with 0.5 mg/l BA.
Elongated shoots rooted on half-strength MS medium supplemented with 0.5 mg/l NAA. Cytological analysis of embryogenic callus
and somatic embryos revealed haploids at a frequency of 30% while that of rooted plants showed haploid regenerants at a frequency
of 8.3%. Nevertheless, the frequency of putative haploid plants could be enhanced through mass multiplication using nodal
explants of the regenerants.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
13.
The effect of in planta TIBA and L-proline onin vitro seedlings and cell culture of sugar beet was investigated. Sterilized seeds were grownin vitro on 1/2 MS medium supplemented with 0 or3 mg/l TIBA. Calli obtained on young leaves cultured on MS medium containing 1 mg/l
BAP, were used for the initiation of cell suspension cultures using MS basal composition supplemented with 0 or 50 mM proline.
Aliquots of 1 ml from cell suspension culture were inoculated onto the first somatic embryo induction MS medium containing
TIBA 0.5 mg/l, BAP 1.0 mg/l, and proline at 0 or 50 mM. After three weeks of culture, embryogenic calli were transferred to
the second embryo induction medium supplemented with NAA and BAP at 0.2 and 0.5 mg/l, respectively. The frequency of somatic
embryos of calli obtained from in plantaTIBA together with proline treatments on average was20 which was higher than that of the other treatments.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
14.
Two different protocols for in vitro regeneration of cassava using zygotic embryos and nodal axillary meristems have been developed. In both cases, buds were regenerated directly from excised explants without an intervening callus phase after a two-step culture procedure. In cotyledonary explants derived from zygotic embryos, prolific shoot formation occurred within 2—3 weeks on MS medium supplemented with 0.5—5 mg/1 BAP alone or in combination with 0.1 mg/1 NAA. Nodal explants with axillary meristems derived from aseptically grown seedlings or stem cuttings were used to initiate a round compact bulb-like structure on MS medium containing 10 mg/1 BAP. These latter structures, when cultured on MS medium supplemented with 0.1 mg/1 NAA, 1 mg/1 BAP and 0.1 mg/1 GA3, produced multiple shoots. Somatic embryos isolated at the globular/torpedo stage from zygotic embryo explants were also capable of multiple shoot production on medium with 1 mg/1 BAP. Rooting of regenerated shoots exceeded 95 % in phytohormone-free MS medium. No change in their ploidy levels was observed. Therefore, the protocols developed should be of use in the particle gun and Agrobacterium-mediated genetic transformation of cassava. 相似文献
15.
本研究以迷迭香叶片为外植体,探索愈伤组织形成及再分化条件。结果表明:蔗糖含量较高的培养基可促进愈伤组织形成,其中以MS+蔗糖50g/L+6-BA0.5mg/L+NAA0.5mg/L效果最好,诱导率可达88%;愈伤组织再分化形成不定芽时,以MS+6-BA1.5mg/L+KT0.5mg/L+NAA0.5mg/L效果较好,再分化率达50%;MS+6-BA0.8mg/L+NAA0.5mg/L诱导不定芽增殖,增殖率可达到300%多;不定芽生根时,MS+NAA0.1mg/L效果较好,生根率可达65%。同时,研究发现,尽管外植体被消毒至无菌,但75%乙醇复合其它灭菌剂共同灭菌时,会导致外植体大量死亡。 相似文献
16.
A protocol for in vitro induction of tetraploids via colchicine-treated somatic embryos from immature zygotic embryos of diploid grapevine (Vitis vinifera L.) is reported. Embryogenic callus was initiated from immature zygotic embryos cultured on Nitsch and Nitsch (NN) medium supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). The callus was transferred to NN medium containing 1.0 mg/l α-naphthalene acetic acid (NAA) and 0.5 mg/l benzyladenine (BA) to establish somatic embryogenesis. The vigorously growing globular embryos were selected and treated by 0, 10 or 20 mg/l colchicine for 1, 2 or 3 days, and then immediately transferred to NN medium supplemented with 0.03 mg/l NAA and 0.5 mg/l BA, for somatic embryo conversion and plant regeneration. The number of surviving embryos and regenerated plantlets following colchicine treatment decreased with increasing colchicine concentration and treatment time. Among 29 randomly investigated plantlets regenerated from colchicine-treated somatic embryos, five solid tetraploids (2n = 4× = 76) were identified by chromosome counting analysis; all others were diploid (2n = 2× = 38). Ploidy level of plant regenerated was also determined from leaves using flow cytometry. No chimeras with both 2C and 4C nuclei was produced from colchicine-treated somatic embryos. Significant differences in leaf stomata parameters were observed between diploid and induced tetraploid plantlets. 相似文献
17.
B. S. Ahloowalia 《Euphytica》1987,36(2):659-665
Summary Plants were regenerated from callus cultures initiated from immature embryos of barley, Hordeum vulgare L. Immature embryos from seven diverse genotypes were cultured on modified Murashige and Skoog (MS) medium supplemented with 1.5 mg 2,4-D and 6.5 mg IAA/l. Of the 249 embryos cultured, 30% initiated callus within 8 days. Subculture of callus for 80 to 100 days on half-MS medium supplemented with 0.5 mg/l 2,4-D and 1.0 mg/l zeatin resulted in organogenesis. Culture of organogenic calli for 30 days on half-MS medium without growth regulators produced plants which originated mostly via multiple shoot formation. Callusing response of the tested genotypes ranged from zero to 44%; however, only 23% of the calli were regenerative. Regenerated plants included variants for chlorophyll deficiency, plant height, stem thickness, spike shape, pollen fertility, seed set and ploidy. 相似文献
18.
冬花椰菜的小孢子胚胎发生研究 总被引：1，自引：0，他引：1
应用花药培养技术在四个冬花椰菜(Brassica oleracea L.ssp.botrytis)基因型中获得小孢子胚胎.培养基中加入硝酸银能明显促进胚胎发生.在含125mg/1硝酸银的培养基上胚产量最高.培养基中加入0.5g/1活性碳,能提高小孢子胚胎发生频率.另外在培养基中用琼脂糖代替琼脂,也能提高胚产量. 相似文献
19.
20.
Brent Tisserat 《Euphytica》1982,31(1):201-214
Summary A discussion of a suitable procedure to rapidly propagate free-living date palms (Phoenix dactylifera L.) from callus cultures is presented. Embryogenic callus derived from lateral bud explants was subjected to various auxin treatments in liquid and agar media including p-chlorophenoxyacetic acid. -naphthaleneacetic acid or 2,4-dichlorophenoxyacetic acid at 0.0, 0.1, 1.0 and 10.0 mg/l concentrations in order to obtain optimum growth. Subsequent plantlet initiation from callus was found to be related to initial auxin pretreatments upon subculture to medium devoid of hormones. Plantlet production from callus subcultured from media containing 0.0, 1.0 and 10.0 mg/l auxin concentrations was notably lower than from callus precultured on the 0.1 mg/l auxin levels. In order to improve in vitro adventitious rooting isolated plantlets were cultured on media containing 0.0, 0.1, 1.0 and 10.0 mg/l concentrations of indole-3-acetic acid or -naphthaleneacetic acid in various physical environments. Optimum adventitious rooting responses and survival in free-living conditions were obtained by culturing plantlets in medium containing 0.1 mg/l for 8–16 weeks prior to transplanting to soil. Axillary shoot outgrowths (offshoots) were found to be common in plantlets cultured on a variety of media once an adequate root-shoot system was developed. Mention of a trademark name or proprietary product does not constitute a guarantee or warranty of the product by the US. Department of Agriculture and does not imply its approval to the exclusion of other products that may be suitable. 相似文献