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1.
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
2.
叶片的生理状态及添加物对大麦叶肉原生质体培养的影响 总被引：1，自引：0，他引：1
作为原生质体源的叶片的生理状态及培养基中的添加物对大麦叶肉原生质体培养均有明显的影响。试验中观察到叶片中、上段的原生质体比基部的具较强的感应力。叶片经黑暗预处理后，其原生质体培养时呈较好的稳定状态，并观察到添加１．０，２．０ｍｍｏｌ／Ｌ的抗坏血酸能提高原生质体的活力。在添加０．５ｍｇ／Ｌ２，４－Ｄ、１．０ｍｇ／ＬＮＡＡ及０．５ｍｇ／ＬＺＴ的ＲＭＳ培养基中，经微弱光照（１５０ｘ）诱导了细胞持续分裂并 相似文献
3.
Daniel S. Moura Francisco J. Zapata-Arias Akihiko Ando Augusto Tulmann Neto 《Euphytica》1997,94(1):01-05
A plant regeneration system from rice protoplasts using calli derived from mature embryos was established for the two Brazilian
modern rice cultivars IAC-201 and IAC-165. After 30 to 40 days of in vitro culture it was possible to obtain on average 6
million protoplasts per gram of callus. Microscopic selection of embryogenic calli was a key step for protoplast isolation.
The production of embryogenic calli increased when L-proline and casein hydrolysate were used in the callus induction medium.
The Oc or IR52 nurse cell lines were essential for protoplast division. Different regeneration media were studied and 139
plants were regenerated which set seed. Some of the regenerated plants showed morphological variation such as the presence
of awns in spite of the short time of the in vitro culture.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
4.
Protoplasts were isolated from leaves of Lotus corniculatus and Trifolium repens and from cotyledons of L. corniculatus. These protoplasts divided and produced colonies. The plating efficiency of protoplasts of both species was improved when agarose was used as a supporting medium. Plants were regenerated more regularly and in larger numbers from colonies of L. carniculatus than T. repens. The use of a culture line of T. repens that had been selected for its response in culture markedly increased the proportion of protoplast-derived cultures which regenerated shoots. One regenerant of T. repens (P6) was analysed for morphological and cytalogical variation. This plant was abnormal and highly aneuploid with a wide range of chromosome numbers. 相似文献
5.
Plants were regenerated from intergeneric somatic hybridization between embryogenic protoplasts of Microcitrus papuana Swingle and leaf-derived protoplasts of sour orange (Citrus aurantium L.) via electrofusion. The regenerated plants were morphologically similar to the leaf parent in growth vigor, leaf and branch
structure. FCM analysis showed that they were diploids. Simple-sequence-repeat (SSR) and cleaved-amplified-polymorphic-sequence(CAPS)
were employed for hybridity characterization. SSR banding patterns of the regenerated plants were identical to the leaf parent,
sour orange, indicating that they possessed nuclear component derived from sour orange. DNA amplification with chloroplast
and mitochondrial universal primers, followed by restriction endonuclease digestion, revealed polymorphism between the fusion
parents. Therefore, this method was used to determine the cytoplasmic compositions of the regenerated plants. Banding patterns
for all the polymorphic primer/enzyme combinations of the regenerated plants were similar to those of the embryogenic parent,
M. papuana, suggesting that only the cytoplasmic components derived from the embryogenic parent were present in the regenerated plants.
FCM, SSR and CAPS demonstrated that intergeneric diploid cybrids have been successfully obtained by symmetric fusion. Related
results concerning nuclear and cytoplasmic composition of previous diploid somatic hybrids and potential mechanism for regeneration
of such kind of plants are discussed herein.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
6.
Summary Calluses of spring and winter wheats (Triticum aestivum L.) were selected for Fusarium resistance in vitro, using the double-layer culture technique. Potato-dextrose agar medium in vials was inoculated with mycelia of Fusarium graminearum and F. culmorum. After one week, fungal cells were killed by autoclaving and the agar medium containing the thermostable toxic metabolites was overlayered with MS callus-growing medium. Later, wheat calluses were placed on the upper medium for 4–5 weeks, and from the surviving calluses plants were regenerated. R2 seedling populations from self-fertilized R1 plants of 4 varieties were tested for Fusarium resistance by artificial infections in the greenhouse, and 3% of the regenerated R2 plants have been found to be more resistant than the original cultivars. 相似文献
7.
Somatic hybrids were obtained from electrofused protoplasts derived from embryogenic suspension cultures of tetraploid cotton (G. hirsutum L. cv. Coker 201) and embryogenic callus of diploid wild cotton G. davidsonii. The regenerants were initially identified as hybrids by RAPD (random amplified polymorphic DNA) analysis. Subsequently, observation on chromosome counting, morphology and SSR (simple sequence repeat) confirmed the hybrid status. Cytological investigation of the metaphase root-tip cells of the regenerated plants revealed there were 74 to 84 chromosomes in the plants, close to the expected 78 chromosomes. SSR analysis revealed the regenerated plants contained specific genomic fragments from both fusion partners, further confirmed their hybridity. The morphology of the plants was intermediate between the two fusion partners. The regenerants were difficult to develop into mature plants because their roots browned and they wilted from the stem apex before forming 3 to 5 true leaves. The hybrid plants were transferred to soil by grafting in vitro onto rootstocks. 相似文献
8.
甘薯和Ipomoea lacunosa的种间体细胞杂种植株再生及鉴定 总被引：10，自引：0，他引：10
用ＰＥＧ融合法融合甘薯品种高系１４号和近缘野生种Ｉｐｏｍｏｅａ ｌａｃｕｎｏｓａ的原生质体，将融合原生质体培养在含有０．０５ｍｇ／Ｌ２，４－Ｄ和０．５ｍｇ／Ｌ ＫＴ的ＭＳ培养基上，愈伤组织迅速增殖。将其中的７０个愈伤组织培养在添加３．０ｍｇ／Ｌ ＢＡＰ的ＭＳ培养基上，并进一步培养在ＭＳ基本培养基上，获得９株再生植株。过氧化物酶同工酶、酯酶同工酶和ＲＡＰＤ分析表明，其中２株再生植株（ＫＬ１和ＫＬ３） 相似文献
9.
G. Spangenberg Z. Y. Wang G. Legris P. Montavon T. Takamizo R. Pérez-Vicente M. P. Vallés J. Nagel I. Potrykus 《Euphytica》1955,85(1-3):235-245
Summary Intergeneric symmetric and asymmetric somatic hybrids have been obtained by fusion of metabolically inactivated protoplasts from embryogenic suspension cultures of tall fescue (Festuca arundinacea Schreb.) and unirradiated or 10–500 Gy-irradiated protoplasts from non-morphogenic cell suspensions of Italian ryegrass (Lolium multiflorum Lam.). Genotypically and phenotypically different somatic hybrid Festulolium mature flowering plants were regenerated.Species-specific sequences from F. arundinacea and L. multiflorum being dispersed and evenly-represented in the corresponding genomes were isolated and used for the molecular characterization of the nuclear make-up of the intergeneric, somatic Festulolium plants recovered. The irradiation of Italian ryegrass protoplasts with 250 Gy X-rays prior to fusogenic treatment favoured the unidirectional elimination of most or part of the donor chromosomes. Irradiation of L. multiflorum protoplasts with 500 Gy produced highly asymmetric (over 80% donor genome elimination) nuclear hybrids and clones showing a complete loss of donor chromosomes.The RFLP analysis of the organellar composition in symmetric and asymmetric tall fescue (+) Italian ryegrass regenerants confirmed their somatic hybrid character and revealed a bias towards recipient-type organelles when extensive donor nuclear genome elimination had occurred.Approaches aimed at improving persistence of ryegrasses based on asymmetric somatic hybridization with largely sexually-incompatible grass species (F. rubra and Alopecurus pratensis), and at transferring the cytoplasmic male sterility trait by intra- and inter-specific hybridization in L. multiflorum and L. perenne, have been undertaken.Abbreviations cpDNA
chloroplast DNA
- CMS
cytoplasmic male sterility
- 2,4-D
2,4-dichlorophenoxy-acetic acid
- IOA
iodoacetamide
- mtDNA
mitochondrial DNA
- RFLP
restriction fragment length polymorphism 相似文献
10.
Summary Three callus initiation media, B2-k, B2, and 7951, were used to study the effects of kinetin on callus initiation, morphology, histology, and regenerability in alfalfa (Medicago sativa L.). The presence of kinetin in callus initiation media retarded callus initiation, but enhanced division and differentiation of callus cells. Calluses induced on kinetin-containing media (B2 and 7951) had many compact cell aggregations, which were considered meristematic regions that might differentiate to plantlets on a regeneration medium. Visually, these calluses were compact and had many nodular structures. In contrast, most calluses induced on a kinetin-free medium were composed of large, individual cells and had friable structures without nodules. After transfer to a hormone-free medium, calluses induced on kinetin-containing media regenerated more frequently than those induced on a kinetin-free medium, but cytokinin (kinetin) autotrophism also occurred. Autotrophism was sexually transmissable and especially affected by the female parent. 相似文献
11.
苹果原生质体培养再生愈伤组织 总被引：2，自引：0，他引：2
以苹果试管苗叶片为原生质体分离材料,对影响原生质体分离和培养的因素进行了研究。结果表明适合叶片酶解的酶液组成是Cellulase-Onzuka R-10 0.8% + Pectinase 0.5% + PVP 1% + 甘露醇 0.65 mol/L + MES0.1%;以改良MT + BA 1.0 mg/L + 2,4-D 0.2 mg/L + 甘露醇0.65 mol/L + Vc 5.0 mg/L + Glu 500 mg/L + CH 100 mg/L + ME 500 mg/L + Arg 50 mg/L为培养基对原生质体进行培养,固液双层培养效果较好,最适培养密度为1×105 (个/ml),培养1-2 d原生质体变形,3-4 d第一次分裂,2 w分裂3-5次。平邑甜茶原生质体一个月后形成微细胞团,两个半月形成肉眼可见的微愈伤组织。鲁加5号和M7均只形成7-10个细胞的细胞团,嘎拉未见细胞分裂。 相似文献
12.
Formation of embryo autonomy of strawberry, plant regeneration fro membryo components, plant freezing conditions in vitro and the possibility to differentiate objectively genotypes by freezing them in vitro and in vivo were studied to create strawberry screening technology in vitro for cold resistance. It was established that autonomy of strawberry embryos manifests itself not earlier than on 14–16th day after pollination and full autonomy is reached on 20–22nd day. Plants regenerated from 26 days old embryos grew most intensively. At the highest rate strawberry plants regenerated
from an isolated embryo axis on MS medium without phytohormones, and from rescued cotyledons x on the medium with 1.0 BA and
0.5 NAA. The temperature interval, at which genotypes differentiated according to cold resistance in vitro, was -8 to 12 °C. Differentiation of strawberry genotypes according to this character conformed to their differentiation in vivo, provided hardening proceeded not less than 21 days. The correlation between cold resistance in vitro and in vivo reached 0.93. Domination of cold resistance manifested itself in strawberry seedlings from various crossing combinations.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
13.
Leaf mesophyll protoplasts of Medicago arborea (2n = 32) were electrofused with cell suspension and/or callus protoplasts of Medicago sativa. Heterokaryons were identified in agarose beds by their dual fluorescein isothiocianate—chlorophyll fluorescence and their coordinates were recorded. Hybrid minicalli were manually picked up and grown first in nurse culture and then in callus induction medium. Hybrid nature of the selected calli was confirmed by isoenzyme analyses. In order to verify whether morphogenesis of somatic hybrid calli was affected by cell incompatibility, mesophyll and cell suspension protoplasts, derived from the same plant of M. sativa with high embryogenic capacity, were fused. Only callus tissues derived from mesophyll protoplasts retained the highly embryogenic character of the M. sativa genotype, while hybrid cell lines were non-morphogenic and showed isoenzyme patterns similar to tissues derived from cell suspension protoplasts. The achievement of somatic hybrid plants in the genus Medicago is discussed. 相似文献
14.
F. J. Stadelmann B. Boller G. Spangenberg R. Kölliker M. Messerli Z. Y. Wang J. Nösberger 《Plant Breeding》1998,117(1):37-43
The effect of regeneration of Lolium perenne and Festuca rubra from embryogenic suspension cells and protoplasts on fertility and growth was evaluated. Embryogenic suspension cultures were either routinely subcultured or cryopreserved and re-established. Phenology, morphology and fertility of regenerated plants were studied for two growing seasons in a replicated field experiment. Most regenerated L. perenne and F. rubra plants showed a delay in inflorescence emergence, a reduced seed yield and differences in morphological traits when compared with seed-grown plants. For L. perenne, performance of plants regenerated from cryopreserved suspension cultures and protoplasts was similar to that of respective plants regenerated from routinely maintained suspension cultures. However, differences in performance were observed for respective regenerants in F. rubra. The phenotypic deviation observed was partly reflected in the randomly amplified polymorphic DNA (RAPD) analysis performed. However, regenerants of both species showing similar, or even superior performance to the seed-grown plants were also found. Embryogenic suspension cells and corresponding protoplasts of L. perenne and F. rubra have the potential for producing fertile, well-performing plants which can be integrated in breeding programs. 相似文献
15.
Of 3272 plants regenerated from protoplasts of 10 Saintpaulia ionantha genotypes, 98.4% survived transfer to the greenhouse. The frequency of regenerants with chlorophyll deficiencies, i.e. variegated leaves or albinos, was low (1.5%). There was a higher number of polyploid, in most cases tetraploid plants, regenerated from protoplasts (16%) which were identified by their altered morphology. Measurements of stomatal length and counting the number of chloroplasts per guard cell also allowed a clear differentiation between diploid and polyploid plants. The classification was confirmed by DNA content determination using flow cytometry. Mechanisms leading to polyploidization included spontaneous protoplast fusion as well as chromosome doubling during callus growth and shoot regeneration. Two genotypes with instabilities in flower colour showed completely altered flower colours in plants regenerated from protoplasts as well as in plants regenerated on leaf explants in vitro. 相似文献
16.
Summary Protoplasts of three Rosa cultivars were fused with each other, with protoplasts of Prunus `Colt' and with protoplasts of Rubus laciniatus, using polyethylene glycol 4000 as a fusogen. Protoplasts of Prunus were incapable of cell division and those of Rosa and Rubus were disabled by treatments with metabolic inhibitors, either iodoacetate (IOA) or rhodamine 6-G (R6G). Parental protoplasts
were then fused in combinations that required complementation for their survival. RAPD analysis of 41 fusion-derived cell
lines showed that two lines resulting from fusions of Rosa + Rosa and one from a fusion of Rosa + Prunus, contained some DNA markers from both fusion partners. The others contained markers of only one fusion parent. This showed
that after protoplast fusion, the heterokaryons did not develop into cell lines with stable hybrid nuclei. Plants regenerated
from cell lines derived from Rosa + Prunus and Rosa + Rubus fusions contained DNA markers of only Rosa and their DNA amounts were no greater than that of the Rosa parent. However, they differed morphologically from the Rosa parent to a remarkable degree, possibly because they inherited undetected genes of Prunus or Rubus, or because they were somaclonal variants of the Rosa parent. Alternative strategies for the production of somatic hybrids are discussed. 相似文献
17.
Multiple shoots were efficiently regenerated from cotyledonary node and shoot tip explants of Pisum sativum within 15 days on MS medium containing B5 vitamins and supplelmented with 2.0 mgl-1 6-benzylaminopurine. The elongated shoots produced on the same medium were excised and transferred to MS medium containing
half strength ammonium nitrate (8.25 gml-1) and supplemented with auxins (indole-3-butyric acid or naphthalene acetic acid) either alone or in combinations with gibberellic
acid. Rooting and flowering were observed on the 7th and 15th day after their transfer to rooting medium. The flowers self-fertilised
in vitro and produced mature pods within 25 days of rooting. These seeds were germinable both in vitro and in vivo. In vitro seeds sown in pots under field conditions developed into flowering plants, and subsequently produced pods with viable seeds.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
18.
Protoplasts from cell suspension cultures of ‘Bonnaza’ navel orange (Citrus sinensis (L.) Osbeck) were electrically fused with mesophyll protoplasts isolated from seedless ‘Red Blush’ grapefruit (Citrusparadisi). After 6 months of culture, a total of 20 plants were regenerated. Root tip chromosome counting revealed that 4 of them
were tetraploids (2n = 4x = 36)and the rest were diploids (2n = 2x = 18) morphologically resembling the mesophyll parent.
After 6 months of transplantation into the greenhouse, 4 of the diploidmesophyll regenerants unexpectedly flowered, but this
phenomenon disappeared in the next year. This is the first report of precocious flowering in citrus via protoplast fusion.
RAPD analysis further confirmed that the tetraploid regenerants were somatic hybrids while the diploid regenerants were mesophyll
parent type. This somatic hybrid will be utilized as a possible pollen parent for improving the seedy pummelo cultivars in
China by producing triploid seedless pummelo hybrid. The mechanism of early flowering was also discussed.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
19.
Using three varieties of Brassica rapa, cv. Hauarad (accession 708), cv. Maoshan-3 (714) and cv. Youbai (715), as the maternal plants and one variety of B. oleracea cv. Jingfeng-1 (6012) as the paternal plant, crosses were made to produce interspecific hybrids through ovary culture techniques. A better response of seed formation was observed when ovaries were cultured in vitro at 9–12 days after pollination on the basal MS and B5 media supplemented with 6-benzylaminopurine (BA) and naphthylacetic acid (NAA). The best response was observed for cross 714×6012 with the rate of seeds per ovary reaching 43.0%. Seeds for cross 715×6012 showed the best germination response (66.7%) on the regeneration medium (MS+1.0 mg l–1 BA+0.05 mg l–1 NAA). In all three cross combinations, good response in terms of root number and length of plants was observed on the root induction medium (MS+1.0 mg l–1 BA+0.1 mg l–1 NAA). A better response was observed for the regenerated plants cultured for 14 days than for 7 days. The ovary-derived plants with well-developed root system were hardened for 8 days and their survival rate reached over 80%. Cytological studies showed that the chromosome number of all plants tested was 19 (the sum of both parents), indicating that these regenerated plants were all true hybrids of B. rapa (n = 10) × B. oleracea (n = 9). The regenerated plants were doubled with colchicine treatment, and the best response in the crosses 708×6012, 714×6012 and 715×6012 was observed when treated with 170 mg l–1 colchicine for up to 30 h and their doubling frequency reached 52, 56 and 62%, respectively. 相似文献
20.
Summary This article reports the culture and plant regeneration of Tripsacum dactyloides. Mature embryos of Tripsacum dactyloides dactyloides were used to obtain embryogenic callus cultures. Currently, 180 normal plants have been regenerated from these cultures. Callus was initiated on MS medium supplemented with dicamba (10 mol or 20 mol) and sucrose (3% or 6%), and plants were regenerated on hormone free MS medium containing 2% sucrose. No significant differences were found in callus initiation frequency or in embryogenic response of cultures on the four combinations of sucrose and dicamba tested. The embryogenic cultures have been maintained for 9 months (12 subcultures) and have retained regeneration capacity. Plants regenerated from tissue culture of maize-by-Tripsacum hybrids could be useful in maize improvement. 相似文献