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1.
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
2.
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
3.
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
4.
Seed dormancy mechanisms in warm season grass species 总被引：6，自引：0，他引：6
Embryogenic protoplasts of Dancy tangerine (Citrus reticulata Blanco) were X-ray irradiated at three doses and electrofused with iodoacetic acid-treated embryogenic protoplasts of Page
tangelo (C. reticulata Blanco × C. paradisi Macf.). Shoots could regenerate only from the fusion combination with the lowest irradiation dose, but were recalcitrant
to rooting. In vitro grafting was applied to obtain complete plants. Chromosome examination showed that the plants contained mainly diploid and
aneuploid cells, together with few tetraploid cells, indicating that they were mixoploids. Random amplified polymorphic DNA
analyses with 10-mer arbitrary primers confirmed the plants as true somatic hybrids. This is the first report on regeneration
of mixoploid hybrid plants via protoplast asymmetric fusion in Citrus. Negative effects of ionizing irradiation on regeneration of embryoids and plantlets and possible agronomic interest of the
mixoploid plants are also discussed.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
5.
V. Orbović M. Ćalović Z. Viloria B. Nielsen F. G. Gmitter Jr. W. S. Castle J. W. Grosser 《Euphytica》2008,161(3):329-335
Five populations of lemon plants [Citrus limon (L.) Burm] obtained from undeveloped ovules through different tissue culture procedures were examined for the presence of
somaclonal and irradiation-induced genetic variation. Tested groups were: (1) nucellar seedlings; (2) organogenic, regenerated
via adventitious buds from nucellar seedling internodes; (3) embryogenic population, regenerated from non-irradiated nucellar
callus via somatic embryogenesis; (4) embryogenic population, regenerated from irradiated nucellar callus via somatic embryogenesis;
and (5) protoplast-derived, regenerated via somatic embryogenesis. Genomic DNA samples from 360 plants (72 from each group)
were screened for polymorphism among RAPD fingerprints amplified by 10 decamer primers. Among all tested plants, genetic variation
was detected only within the group of plants recovered from irradiated embryogenic calli. Out of 72 plants from that group,
three had RAPD fingerprints different from the rest of the population, and fourth plant was found to be cytochimeric, consisting
of diploid and tetraploid cells as revealed by flow cytometry. In all other populations of regenerated plants, we did not
come across any plants with changed ploidy level. 相似文献
6.
The aim was to develop an efficient chromosome doubling method for Miscanthus sinensis to enable the production of triploids and so avoid seed dispersal to the environment. Antimitotic treatments with colchicine or oryzalin were tested in M. sinensis cl. MS‐88‐110 on: (1) in vitro shoots and plants established in soil; (2) during propagation of embryogenic callus; and (c) during the initial stages of callus induction. All systems produced chromosome‐doubled plants. A higher percentage of tetraploids was found after antimitotic treatment at the explant or callus level compared with treatment of in vitro shoots or plants established in soil. In general, oryzalin was more toxic to plant material than colchicine. A higher frequency of chimeras was found among plants with altered ploidy level when the target was formed shoot buds compared with adventitious shoot formation from callus. Antimitotic treatment of embryogenic callus from shoot apices also resulted in a high degree of albinism. 相似文献
7.
D. Sihachakr R. Haïcour J.M. Cavalcante Alves I. Umboh D. Nzoghé A. Servaes G. Ducreux 《Euphytica》1997,96(1):143-152
The application of new techniques for improvement of sweet potato crops, particularly including the exploitation of somaclonal
variation, gene transfer by genetic transformation and somatic hybridization, requires the control of plant regeneration from
tissue cultures. Shoots can easily be regenerated from explants of stems, petioles, leaves and roots, while callus cultures
do not produce any shoots. The potential of somatic embryogenesis and plant regeneration via embryogenesis was evaluated for
10 cultivars of sweet potato. Protocols for plant regeneration from cultured protoplasts have also been developed. Since mesophyll
was resistant to enzyme digestion, fragments of stems and petioles, callus and cell suspensions were used as source of protoplasts
of sweet potato. Series of transfers of protoplast-derived calluses, particularly those which had been obtained from in vitro
plants, to media containing a high level of zeatin resulted in successful formation of shoots in only two sweet potato cultivars.
In addition, the embryogenic potential was irreversibly lost through protoplast culture, since protoplasts isolated from embryogenic
cell suspensions developed into non-embryogenic callus. Consequently, an alternative protocol is being successfully developed
to improve plant regeneration from cultured protoplasts of sweet potato, involving first root formation from which shoots
can then be regenerated. Preliminary evaluation in field conditions in Gabon revealed that plants regenerated from cultured
protoplasts exhibited a great genetic variability in their growth and tuber formation in particular.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
8.
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
9.
Somatic embryogenesis (SE) is a critical step leading to plant regeneration in tissue culture of many plant species. The objective
of the present study was to analyze the inheritance of SE in cotton (Gossypium hirsutum L.) using leaf petioles as explants. A high embryogenic callus (HEC)—producing line, W10, was selected by petiole callus
culture from a commercial Chinese cotton cultivar CRI24 and crossed with a non embryogenic line, TM-1 and a low embryogenic
(LEC) commercial Chinese cotton cultivar, CRI12, respectively. The parental lines, F1 and F2 were grown in field conditions for sources of leaf petioles as explants. The F1 plants were similar to the HEC parent in embryogenic callus (EC) induction, indicating that high EC ability is dominant.
The classical Mendelian analysis showed that the high EC ability in the HEC line W10 is controlled by two independent dominant
genes with complementary effect, designated Ec
1
and Ec
2
, while the LEC line CRI12 contains one dominant gene Ec
2
. A joint segregation analysis confirmed that SE ability in cotton is controlled by two major genes with epistatic effects
along with other polygenes. A SSR marker analysis identified three quantitative trait loci (QTLs) on two linkage groups, one
of which harbored a major QTL (qEc1) which is assigned to the major gene Ec
1
. This qualitative and quantitative genetic study has provided an incentive to fine map the genes responsible for SE towards
the isolation of the SE genes in cotton. 相似文献
10.
为发掘枳橙同源四倍体资源,从而为柑橘砧木遗传改良提供新种质,本研究通过初步的形态学观察,从具多胚性的柑橘重要砧木资源枳橙实生苗群体750个单株中初步筛选出具典型多倍体特征的植株24株,其中流式细胞仪鉴定为枳橙四倍体22株,群体四倍体自然发生率达2.93%。下表皮气孔分析表明枳橙四倍体气孔密度显著低于二倍体对照。对获得的四倍体植株进行了SSR分子鉴定,19对引物扩增结果均显示枳橙四倍体与其二倍体对照的带型完全一致,表明获得的四倍体枳橙均为同源四倍体。本研究表明将形态学初选与流式细胞仪快速鉴定及SSR分析相结合是从多胚性柑橘资源实生群体获得同源四倍体的简便有效途径,本研究发掘的同源四倍体枳橙是柑橘砧木遗传改良相关研究的良好资源。 相似文献
11.
二倍体、四倍体小麦成熟胚组织培养研究初报 总被引：2，自引：0，他引：2
以栽培二粒小麦(Triticum dicoccum Schuble)、硬粒小麦(Triticum durum Desf.)以及普通六倍体小麦祖先种野生二粒小麦(Triticum dicoccoides Kom.)、野生一粒小麦(Triticum aegilopides Bal.)4个基因型麦属植物成熟胚(MEs)为外植体,对成熟胚愈伤组织诱导、分化及植株再生的组织培养效果做了初步研究,筛选出适合于非六倍体小麦成熟胚组织培养的基因型.结果表明,不同基因型成熟胚愈伤组织的诱导、分化及植株再生差异显著,具有很强的基因型效应和基因型依赖性.其中栽培二粒小麦的成熟胚表现出较好的组织培养特性,它的愈伤组织诱导率、分化率、成苗率以及组培效率分别为95.00%、90.00%、32.40%、27.70%,显著高于其它基因型.二倍体、四倍体小麦成熟胚组织培养特性的研究以及组织培养基因型的筛选,为遗传转化体系的建立、小麦功能基因的分离和克隆,以及小麦分子育种工作奠定了基础. 相似文献
12.
摘要∶南丰蜜橘是原产于中国的古老柑橘品种,其果实皮薄而肉甜,深受消费者喜爱,当前南丰蜜橘的品质发生劣变,影响了其商品价值。为采用基因工程育种以改善南丰蜜橘品质,通过诱导其愈伤组织并以其为试材进行了转基因研究。南丰蜜橘自种子实生苗下胚轴处长产生出愈伤组织,挑选胚性愈伤组织进行增殖后,采用根癌农杆菌介导法进行开花相关基因AP1的转化。结果获得22团抗性愈伤组织,平均产出率为7.3团/皿,表明该品种有较高的转化率。抗性愈伤组织经PCR分析、Southern blot检测证实为转化子。本研究结果表明南丰蜜橘胚性愈伤组织是基因工程改良的良好受体,可用于其它基因的转化研究。 相似文献
13.
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
14.
C.E. van Schaik C. van der Toorn M.J. De Jeu C.J.J.M. Raemakers R.G.F. Visser 《Euphytica》2000,115(1):17-26
The successful application of plant biotechnology to Alstroemeria improvement will largely depend on the availability of an efficient regeneration/transformation system. Regeneration in Alstroemeria is accomplished from nodular embryogenic callus initiated from zygotic embryos. Histological studies of embryogenic callus
initiation from 4-weeks old cultured ovules revealed that the outermost layers of the protoderm of the embryogenic nodules
divided to form either a new nodule or aproembryo. Transient gene expression after particle bombardment of nodular embryogenic
callus was optimized using DNA of pAHC25. The highest β-glucuronidase expression was found when the GUS gene was under control of the maize ubiquitin promoter, the target tissue was placed 5 cm below the microcarrier launch assembly
and when the rupture disc-breakage point was between 650–900 psi. Kanamycin blocked regeneration of somatic embryos, however,
did not block growth of nodular embryogenic callus. With phosphinothricin both callus growth and regeneration were blocked.
Bombardment of nodular embryogenic callus with DNA of pAHC25 combined with selection on medium containing phosphinothricin
resulted in putative transgenic chimeric. Friable calli were selected from nodular embryogenic callus and used to initiate
suspensions. These cell suspensions were subjected to transformation by particle bombardment using DNA of pAHC25 and resulted
in a stable transformed friable callus line after selection based on luciferase activity. Even after 2 years of maintenance
this callus line was luciferase positive and the Polymerase Chain Reaction analysis demonstrated the presence of the introduced
gene in this friable callus line.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
15.
The behaviour of diverse Triticum genotypes in the tissue culture response of mature embryo callus was compared, and factors affecting tissue culture response were studied in this paper. Significant differences were detected in callus induction, embryogenic callus differentiation, plantlet regeneration and culture efficiency when mature embryos of 31 plants of different Triticum species were compared. These were the main wheat cultivars of the Chinese northern winter‐type wheat region and breeding lines (Triticum aestivum L.), durum wheat (Triticum durum Desf.), cultivable emmer wheat (Triticumdicoccum Schuble) and the common wheat progenitors Triticum dicoccoides and Triticum aegilopides. The genotype dependency was particularly high in tissue culture of mature embryos of these Triticum genotypes. The efficiency of induction, differentiation and regeneration of mature embryos callus was high in genotypes selected out. Mature embryo‐derived callus of HB341, TS021, SN2618, T. dicoccum, HB188, and T9817 showed better tissue culture response than the other genotypes. Plantlets can be regenerated from mature embryo‐derived callus of 31 genotypes, saving on growth facility resources and time required for the collection of other explants, and providing a solid basis for the genetic transformation and molecular plant breeding of Triticum plants. 相似文献
16.
A protocol was established for high frequency cyclic somatic embryogenesis for different varieties of cassava. An efficient
plant regeneration system was developed for the high cyanogenic variety PRC60a. Linamarin content and linamarase activity
were determined in various tissues of secondary somatic embryos and regenerated plants of PRC 60a. Both linamarin and linamarase
activity were not detected in embryogenic callus, roots induced from callus and somatic embryo tissues. The stems and leaves
of regenerated plants (in vitro) and storage roots and leaves of mature plants (in vivo), however, contained variable amounts of linamarin and linamarase activity whereas in the non storage root tissues (in vitro) only linamarin was detected. The present study suggested that the linamarin biosynthetic pathway may be absent or not switched
on in the embryogenic callus and somatic embryos. The ploidy level and somatic chromosome number of the regenerated plants
were found to be same as the source plants. The availability of this regeneration system would be useful not only for investigating
cyanogenesis but also for genetic manipulation in cassava.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
17.
Production of embryogenic tissues and regeneration of transgenic plants in cassava (Manihot esculenta Crantz) 总被引：3，自引：0，他引：3
Nigel J. Taylor Munyaradzi V. Masona Rosa Carcamo Thao Ho Christian Schöpke Claude M. Fauquet 《Euphytica》2001,120(1):25-34
Disorganised embryogenic tissues have been utilised as target tissues for transgene insertion and transgenic plant regeneration
in cassava (Manihot esculenta). The production of friable embryogenic callus in fourteen geographically diverse cassava cultivars, from which eleven were
established as embryogenic suspension cultures, is reported. Embryogenic tissues were similar in nature in all cultivars tested
although there was variation in the time required to generate friable callus and the growth rates of suspension cultures.
Regeneration of plants has been achieved from eight cultivars but varied significantly in efficiency, with cv. TMS 60444and
Line 2 from Zimbabwe being the most responsive. Tissues from the remaining eight cultivars became arrested at globular and
torpedo stages of regeneration indicating that they most likely process an inherent ability to produce plants but require
further research to allow this to be realised. Significant numbers of transgenic plants containing transgenes for putative
resistance to important viral diseases of cassava in addition to visual marker genes have been regenerated. Transgenic plants
from three the cultivars TMS 60444, Bonoua Rouge and M.Col 1505 were recovered after particle bombardment of embryogenic suspension
cultures. Correlation's have been made between abnormal leaf morphology and plant vigour with the use of embryogenic suspension
cultures for transgene insertion. As an result friable embryogenic callus is now being successfully utilsed as the target
tissue for genetic transformation and plant regeneration at ILTAB.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
18.
The present investigation discusses the scope for transferring of resistance to leaf spot disease incited by Alternaria helianthi from two hexaploid wild species (H. tuberosus and H. resinosus) to diploid cultivated sunflower. Interspecific hybrids produced between sunflower and these two hexaploid species were partially fertile with tetraploid chromosome status. Backcrosses of these interspecific hybrids with cultivated sunflower resulted in the formation of sterile triploid plants. To overcome the problem of sterility and facilitate backcrosses with cultivated sunflower, anther culture of the tetraploid interspecific hybrids was carried out to bring down their chromosome number to diploid status. Anthers from both interspecific hybrids were cultured on basal Murashige and Skoog media supplemented with varying concentrations of organics and the growth regulators benzyladenine and naphthaleneacetic acid. Anthers of interspecific hybrids involving H. resinosus responded well and regenerated through an embryogenic route at a frequency of 98.7%. But in interspecific hybrids with H. tuberosus, anthers formed callus and subsequently regenerated shoots through an organogenic pathway. DNA ploidy analysis of anther culture plants of interspecific hybrids derived from H. tuberosus crosses was carried out to identify plants with desired diploid status. In vitro screening of parents, interspecific hybrids and anther culture plantlets against A. helianthi showed resistance in 68.5% of the anther culture plants of interspecific hybrids from H. tuberosus and in 24.3% of the plants derived from interspecific hybrids involving H. resinosus. 相似文献
19.
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
20.
Chloroplast microsatellite variations in tetraploid alfalfa 总被引：2，自引：0，他引：2
Four chloroplast simple sequence repeat (cpSSR) loci were identified in alfalfa (lucerne), Medicago sativa L. The occurrence of allelic variation at these loci was evaluated in 100 plants from 10 populations of tetra‐ploid alfalfa, belonging to four Italian ecotypes and to an Italian (‘Lodi’) and an Egyptian (‘Iside’) variety. Twenty‐four different alleles were identified, four of which were exclusive to the ‘Iside’ variety. The genetic relationship among plants and among populations was analysed by computing an analysis of molecular variance and an unweighted pair group means analysis clustering. This analysis allowed clear separation of the ‘Iside’ variety from Italian germplasm and the recognition of close relationships within the Italian populations. The data presented suggest that cpSSR analysis of tetraploid alfalfa could be used for germplasm polymorphism analysis. 相似文献