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1.
M. Q. Wang Z. Y. Peng M. C. Wang W. Guo J. S. Zhao D. Y. Zhi G. M. Xia 《Russian Journal of Plant Physiology》2011,58(4):615-621
Protoplasts of Bupleurum scorzonerifolium irradiated with 380 μW/cm2 UV for 5 min were fused by the PEG-mediated method with untreated protoplasts of Arabidopsis thaliana. The fusion products were cultured in the P5 liquid medium for single hybrid cell clone formation. As a total, 81 independent
putative hybrid clones (cell lines) were obtained, and seventeen of them were identified as somatic hybrids by chromosome
counting, GISH, RAPD, and SSR analyses. More than 80 B. scorzonerifolium-like green introgressed plants and leaves were regenerated from 49 somatic hybrid cell lines, which contained chromatin and
DNA characteristic of A. thaliana. To assess the UV tolerance of both parents with chromatin exclusion and introgression, their protoplasts were UV-irradiated
(380 μW/cm2 for 0 and 5 min), and the protoplasts of A. thaliana were more sensitive to UV than those of B. scorzonerifolium as judged by Single Cell Gel Electrophoresis analysis. The possible relationship between UV resistance of B. scorzonerifolium and A. thaliana chromosome elimination and the formation of somatic introgressed hybrid plants is discussed. 相似文献
2.
In our early experiments, a variety of Bupleurum scorzonerifolium-like somatic hybrid plants were obtained from protoplast fusion between Arabidopsis thaliana and UV-treated/untreated B. scorzonerifolium. To compare the effects of UV and γ-ray irradiation on the B. scorzonerifolium partner and obtain Arabidopsis-like hybrids, we designed a novel combination of somatic hybridization between A. thaliana and B. scorzonerifolium. Before protoplast isolation and fusion, the suspension cells of B. scorzonerifolium were irradiated by gamma ray (60Co, 50 Gy with 1.3 Gy min−1). Both parental protoplasts lost regeneration capacity, but over 100 somatic hybrids restored the capacity and developed
to Arabidopsis-like inflorescences and flowers with some characteristics of B. scorzonerifolium. Some hybrid flowers showed yellow sepal, petal, or carpel, whose color was similar to the petal of B. scorzonerifolium; the others had silique of Arabidopsis with angularity of B. scorzonerifolium, and their parts possessed five stamens, the same as B. scorzonerifolium. Cytological analysis showed that three hybrids had Arabidopsis-like karyotypes. Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeats (SSR) profiles revealed that both parental
fragments were amplified from these hybrids. These results indicated chromatin introgression from B. scorzonerifolium to A. thaliana, which may be related to the complementation of hybrid inflorescence and flower generation. 相似文献
3.
M. Wang Z. Peng L. Wang J. Zhao J. Che G. Xia 《Russian Journal of Plant Physiology》2011,58(1):133-141
The protoplasts of tall fescue (Festuca arundinacea Schreb.) were fused with those of Bupleurum scorzonerifolium Willd. The latter were irradiated with UV at an intensity of 380 μW/cm2 for 0 s (combination I), 30 s (combination II), and 60 s (combination III) before fusion. Putative hybrid calli, leaves,
and shoots were generated from the fusion products. They were recognized as somatic hybrids by a combined analysis of chromosome
numbers, isozyme, RAPD, and 5S rDNA spacer sequence. The hybrid calli with morphogenetic ability and leaves/shoots differentiation
had the B. scorzonerifolium phenotype, whether they were derived from symmetric fusion (UV 0 s) or asymmetric fusion (UV 30 s/60 s). Cytological tests
revealed that these hybrids contained the complete set (12) of B. scorzonerifolium chromosomes and 0–4 partner tall fescue chromosomes. The tall fescue chromosomes were rapidly eliminated in combinations
II and III, but gradually lost in combination I. It was noted that the green leaves and shoots were produced earlier, and
the differentiation frequency was higher in combinations II and III than in combination I, which corresponded to the speed
of elimination of the tall fescue chromosomes in the hybrids. Therefore, UV irradiation can indirectly promote elimination
of tall fescue chromosomes and hybrid differentiation. B. scorzonerifolium can repel partner chromosomes with mechanism that differs from UV. 相似文献
4.
Callus-derived protoplasts of common wheat (Triticum aestivum L. cv. Hesheng 3) irradiated with ultraviolet light were fused by using the PEG method with cell suspension-derived protoplasts
of Arabidopsis thaliana. Regenerated calli and green plants resembling that of wheat were obtained. The hybrid nature of putative calli and plants
were confirmed by isozyme, random amplified polymorphic DNA and genomic in situ hybridization (GISH) analyses. GISH results
indicated that 1∼3 small chromosome fragments of A. thaliana were found introgression into the terminals of wheat chromosomes, forming highly asymmetric hybrids. Cytoplasmic genome tests
did not show any cytoplasmic genetic materials from A. thaliana. However, variations from the normal wheat cytoplasmic genome were found, indicating recombination or rearrangement occurred
during the process of somatic hybridization. The chromosome elimination in the asymmetric somatic hybridization of remote
phylogenetic relationship was discussed. A miniature inverted-repeat transposable element related sequence was found by chance
in the hybrids which might accompany and impact the process of somatic hybridization.
Jingyao Deng and Haifeng Cui provided same contribution to this work. 相似文献
5.
In the present investigation, the interspecific somatic hybridization between tuber mustard and red cabbage was established in order to introduce valuable genes from red cabbage (Brassica oleracea) into Brassica juncea. Prior to fusion treatment, protoplasts of red cabbage were inactivated with 2 mM iodoacetamide to inhibit cell division. Micro-calluses were obtained at a frequency of 10.3% after approximately 5 weeks culture following protoplast fusion. Some of the fusion-derived calluses possessed red pigmented cells after being transferred to proliferation medium, and they were presumably considered to be somatic hybrid cell lines. Plantlets were regenerated from 12 cell lines, of which nine plantlets exhibited characteristics intermediate of both parents in terms of plant morphology. With the exception of common protein bands featured by two parents, there were unique banding patterns produced in the hybrids by using SDS-PAGE analysis. By chromosome countings, it was showed that they ranged approximately from 2n=30 to 42 in chromosome numbers. Their hybridity were further confirmed by RAPD analysis revealing that genes of both parents were partially incorporated into the hybrids. Positively, all these hybrids were capable of seed-setting. The pod-setting was 4.2 in somatic hybrid H7 when backcrossed with tuber mustard. 相似文献
6.
Alien chromosome addition lines have been widely used for identifying gene linkage groups, assigning species-specific characters
to a particular chromosome and comparing gene synteny between related species. In plant breeding, their utilization lies in
introgressing characters of agronomic value. The present investigation reports the production of intergeneric somatic hybrids
Brassica
napus (2n = 38) + Orychophragmus violaceus (2n = 24) through asymmetric fusions of mesophyll protoplasts and subsequent development of B. napus-O. violaceous chromosome addition lines. Somatic hybrids showed variations in morphology and fertility and were mixoploids (2n = 51–67) with a range of 19–28 O. violaceus chromosomes identified by genomic in situ hybridization (GISH). After pollinated with B. napus parent and following embryo rescue, 20 BC1 plants were obtained from one hybrid. These exhibited typical serrated leaves of O. violaceus or B. napus-type leaves. All BC1 plants were partially male fertile but female sterile because of abnormal ovules. These were mixoploids (2n = 41–54) with 9–16 chromosomes from O. violaceus. BC2 plants showed segregations for female fertility, leaf shape and still some chromosome variation (2n = 39–43) with 2–5 O. violaceus chromosomes, but mainly containing the whole complement from B. napus. Among the selfed progenies of BC2 plants, monosomic addition lines (2n = 39, AACC + 1O) with or without the serrated leaves of O. violaceus or female sterility were established. The complete set of additions is expected from this investigation. In addition, O. violaceus plants at diploid and tetraploid levels with some variations in morphology and chromosome numbers were regenerated from the
pretreated protoplasts by iodoacetate and UV-irradiation.
Z. Zhao and T. Hu make equal contributions to this work. 相似文献
7.
In this paper, we describe how Bupleurum scorzonerifolium/Triticum aestivum asymmetric somatic hybrids can be exploited to study the wheat genome. Protoplasts of B. scorzonerifolium Willd were irradiated with ultraviolet light (UV) and fused with protoplasts of common wheat (T. aestivum L.). All cell clones were similar in appearance to those of B. scorzonerifolium, while the regenerated plantlets were either intermediate or B. scorzonerifolium-like. Genotypic screening using isozymes showed that 39.3% of cell clones formed were hybrid. Some of the hybrid cell clones grew vigorously, and differentiated green leaves, shoots or plantlets. DNA marker analysis of the hybrids demonstrated that wheat DNA was integrated into the nuclear genomes of B. scorzonerifolium and in situ karyotyping cells revealed that a few wheat chromosome fragments had been introgressed into B. scorzonerifolium. The average wheat SSR retention frequency of the RH panel was 20.50%, but was only 6.67% in fusions with a non-irradiated donor. B. scorzonerifolium chromosomes and wheat SSR fragments in most asymmetric hybrid cell lines remained stable over a period of 2.5–3.5 years. We suggest the UV-induced asymmetric somatic hybrids between B. scorzonerifolium Willd and T. aestivum L. have the potential for use in the construction of an RH map of the wheat genome. 相似文献
8.
Tanveer Khan Vanga Siva Reddy Sadhu Leelavathi 《Plant Cell, Tissue and Organ Culture》2010,101(3):323-330
A highly efficient and reproducible regeneration system based on somatic embryogenesis in Gossypium hirsutum cv. Narasimha (NM), which has superior fiber qualities and is also used as a female parent in several hybrid cottons, has
been developed. Embryogenic callus was obtained form both hypocotyls and cotyledonary leaves on Murashige and Skoog (MS) medium
containing kinetin and 2,4-dichlorophenoxyacetic acid. Somatic embryogenesis was observed on hormone-free MS medium, but embryos
did not grow well beyond globular stage. However, somatic embryos germinated well on MS medium containing B5 vitamins; addition
of zeatin was found to be beneficial for their normal development. Most importantly, the media and culture conditions developed
for NM were also found to be suitable for high-frequency somatic embryogenesis in Coker 310. In addition, the newly developed
regeneration protocol has been successfully tested for genetic transformation through co-cultivation with Agrobacterium using embryogenic calli as explants. Molecular analysis confirmed the stable integration and expression of marker gene, green
fluorescent protein (GFP). These results show that it is now possible to introduce foreign gene(s) directly into elite cultivar
Narasimha with similar efficiency to in traditionally used Coker lines in a relatively short period of time. Development of
efficient regeneration and transformation systems as demonstrated here should augment the introduction of new traits directly
into cultivated varieties/hybrids, reducing the time required for back-crossing and the costs for seed production, besides
aiding genomic research in cotton. 相似文献
9.
Antifreeze proteins depress the non-equilibrium freezing point of aqueous solutions, but only have a small effect on the equilibrium
melting point. This difference between the freezing and melting points has been termed thermal hysteresis activity (THA).
THA identifies the presence and relative activity of antifreeze proteins. Two antifreeze protein cDNAs, dafp-1 and dafp-4, encoding two self-enhancing (have a synergistic effect on THA) antifreeze proteins (DAFPs) from the beetle Dendroides canadensis, were introduced into the genome of Arabidopsis thaliana via Agrobacterium-mediated floral dip transformation. Southern blot analysis indicated multiple insertions of transgenes. Both DAFP-1 and/or
DAFP-4 were expressed in transgenic A. thaliana as shown by RT-PCR and Western blot. Apoplastic fluid from T
3 DAFP-1 + DAFP-4-producing transgenic A. thaliana exhibited THA in the range of 1.2–1.35°C (using the capillary method to determine THA), demonstrating the presence of functioning
antifreeze proteins (with signal peptides for extracellular secretion). The freezing temperature of DAFP-1 + DAFP-4-producing
transgenic A. thaliana was lowered by approximately 2–3°C compared with the wild type. 相似文献
10.
S. Kumar Sunil Kumar S. P. Negi J. K. Kanwar 《In vitro cellular & developmental biology. Plant》2008,44(6):474-479
Callus cultures derived from leaf segments of chrysanthemum cultivar ‘Snow Ball’ which was susceptible to Septoria obesa were successfully used for in vitro selection for resistance to this pathogenic fungus. Resistant cell lines were selected by culturing callus on growth medium
containing various concentrations of S. obesa filtrate. Resistant calluses obtained after two cycles (30 d each cycle) of selection were used for plant regeneration. About
30% of the plants regenerated from the resistant calluses and 70–80% of the plants raised from cuttings had acquired considerable
resistance against the pathogen in the field. No phenotypic variation was observed in the selected regenerates. 相似文献
11.
The chromosome constitution of hybrids and chromatin patterns of Agropyron elongatum (Host)Neviski in F5 somatic hybrid lines -1–3 and I-1-9 between Triticum aestivum L. and A. elongatum were analyzed. Based on the statistic data
of pollen mother cells, F5 I-1-9 and-1-3 had 20–21 bivalents with a frequency of 84.66% and 85.28%, of which, 89.83% and 89.57% were ring bivalents.
The result indicated that both hybrid lines were basically stable in the chromosome constitution and behavior. RAPD analysis
showed that the two hybrids contained biparental and integrated DNA. GISH (Genome in situ hybridization) revealed that in
the form of small chromosome segments, A. elongatum chromatin was scattered on 4–6 wheat chromosomes near by the region of
centromere and telomere in the two hybrid lines. SSR analysis indicated that A. elongatum DNA segments were distributed on
the 2A, 5B, 6B and 2D wheat chromosomes in the hybrids, which was in accordance with the GISH results that small-segments
intercalated poly-site. 相似文献
12.
Karin Sonntag Brigitte Ruge-Wehling Peter Wehling 《Plant Cell, Tissue and Organ Culture》2009,96(3):297-305
A method for isolation and shoot regeneration from electrofused protoplasts of L. angustifolius and L. subcarnosus was developed. Viable protoplasts were isolated from leaves of in-vitro grown seedlings at an average yield of 6 × 105 protoplasts g−1 fresh weight. Liquid and agarose solidified B5 media were used for protoplast culture. In the liquid-culture system, all
tested media, VKM, P1 and KM8p, were applicable for inducing cell division (84% of all tested petri dishes at four weeks)
and colony formation. Media containing additional carbohydrates were suitable to produce compact calli with green and brown
pigmentations in different combinations. Analysis of callus with molecular markers allowed to identify six somatic hybrids.
However, none of the parental-protoplast derived cell colonies could develop shoots. This is the first report on protoplast
fusion of L. angustifolius and L. subcarnosus with subsequent shoot regeneration. 相似文献
13.
Anber Hassanein Latifa Hamama Karine Loridon Noëlle Dorion 《Plant cell reports》2009,28(10):1521-1530
Direct genetic transformation of mesophyll protoplasts was studied in Pelargonium × hortorum. Calcein and green-fluorescent protein (GFP) gene were used to set up the process. Electroporation (three electric pulses
from a 33-μF capacitor in a 250-V cm−1 electric field) was more efficient than PEG 6000 for membrane permeation, protoplast survival and cell division. Transient
expression of GFP was detected in 33–36% of electroporated protoplasts after 2 days and further in colonies. A protoplast
suspension conductivity of >1,500 μS cm−1 allowed high colony formation and plant regeneration. Stable transformation was obtained using the plasmid FAJ3000 containing
uidA and nptII genes. When selection (50 mg l−1 kanamycin) was achieved 6 weeks after electroporation, regenerated shoots were able to grow and root on 100 mg l−1 kanamycin. The maximum transformation efficiency was 4.5%, based on the number of colonies producing kanamycin-resistant
rooted plants or 0.7% based on the number of cultured protoplasts. Polymerase chain reaction (PCR) analysis on in vitro micropropagated
plants showed that 18 clones out of 20 contained the nptII gene, while the uidA gene was absent. These results were confirmed after PCR analyses of five glasshouse-acclimatized clones. 相似文献
14.
In the present paper, attempts were made to explore the possibility of employing ultraviolet (UV) irradiation in citrus asymmetric
fusion for transfer of limited amount of favorable traits from a desirable cultivar to a target one. Exposure of Satsuma mandarin
(Citrus unshiu Marc.) embryogenic protoplasts to UV at an intensity of 300 μW cm−2 led to reduced viability, especially under long irradiation duration. The protoplasts could not grow during culture when
they were irradiated for over 30 s. Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) assay revealed
extensive DNA fragmentation in the UV-irradiated protoplasts compared with those without UV treatment. Electrofusion between
UV-irradiated protoplasts of Satsuma mandarin (donor) with those of Jincheng (C. sinensis Osbeck, recipient), a local cultivar of superior quality, gave rise to regeneration of several lines of shoots, which failed
to root despite enormous endeavors. Ploidy analysis via flow cytometry and chromosome counting showed that four selected shoots
were either diploid, triploid or tetraploid. Random amplified polymorphism DNA (RAPD) and amplified fragment length polymorphism
(AFLP) confirmed the shoots, irrespective of their ploidy level, as putative somatic hybrids. Cleaved amplified polymorphism
sequences (CAPS) demonstrated that the shoots predominantly got their cytoplasmic components, in terms of chloroplast (cp)
and mitochondrion DNA, from Jincheng, along with possible recombination of cpDNA in some shoot lines. The current data indicated
that UV-based asymmetric fusion could also be employed in citrus somatic hybridization with the intention of creating novel
germplasms, which may provide an alternative approach for cultivar improvement. 相似文献
15.
Lourdes Castelblanque Begoña García-Sogo Benito Pineda Vicente Moreno 《Plant Cell, Tissue and Organ Culture》2010,100(1):107-112
A simple and efficient protocol for plant regeneration from protoplasts of the potted plant Kalanchoe blossfeldiana Poelln. is reported. Mesophyll protoplasts were isolated from axenic leaves after a preculture. The enzymatic digestion of
the tissue with a solution containing 0.4% Cellulase Onozuka R-10 and 0.2% Driselase yielded 6.0 × 105 protoplasts per gram fresh weight after density gradient purification. Protoplasts were cultured in the dark at an initial
density of 1 × 105 protoplasts per milliliter in a liquid medium with 320 mM mannitol, 130 mM sucrose, 2.3 μM 2,4-dichlorophenoxy acetic acid
(2,4-D), 5.4 μM 1-naphthaleneacetic acid (NAA) and 2.2 μM 6-benzyladenine (BA). Cell wall regeneration was observed within
4 days of culture and cell division began after 5–7 days. When cultured in a liquid medium with 5.4 μM NAA and 8.9 μM BA,
protoplast-derived colonies proliferated until small visible calli, and adventitious buds appeared after transfer to photoperiod
conditions. Developed shoots were rooted on a solid medium supplemented with 0.6 μM indole-3-acetic acid (IAA) and successfully
established under greenhouse conditions. The process required 4 months from isolation to rooted plants and the best conditions
found gave a plant regeneration efficiency of 6.4 plants per 1 × 105 protoplasts. This is the first protocol reported for plant regeneration from protoplasts for a Crassulaceae family species. 相似文献
16.
A storage root-bearing somatic hybrid was produced for the first time by protoplast fusion between sweetpotato (Ipomoea batatas (L.) Lam.) cv. Kokei No. 14 and its wild relative I. triloba L. Protoplasts isolated from embryogenic suspension cultures of Kokei No. 14 were fused with petiole protoplasts of I. triloba L. using polyethylene glycol-mediated protocol. Fusion products were cultured in a modified Murashige and Skoog medium containing
0.05 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg l−1 kinetin. A total of 176 plants were obtained from 42 out of 134 calluses derived from fused protoplasts, and 91 of these
plants were confirmed to be somatic hybrids through peroxidase isozyme, random amplified polymorphic DNA, amplified fragment
length polymorphism, and cytological analyses. Upon transfer into soil and grown in the greenhouse and then to the field,
100% survival was observed. A single plant, designated KT1, was found to produce storage roots. Genomic in situ hybridization
analysis confirmed presence of chromosomes from both parents and recombinant chromosomes in KT1. Drought tolerance, dry matter
content, soluble sugar content, and fertility of this somatic hybrid were evaluated for potential use in sweetpotato breeding. 相似文献
17.
Jagesh K. Tiwari Poonam D. Sarkar SK. Pandey Jai Gopal S. Raj Kumar 《Plant Cell, Tissue and Organ Culture》2010,103(2):175-187
Interspecific potato somatic hybrids between Solanum tuberosum L. (di)haploid C-13 and 1 endosperm balance number non-tuberous wild species S. etuberosum Lindl. were produced by protoplasts electrofusion. The objective was to transfer virus resistance from this wild species
into the cultivated potatoes. Post-fusion products were cultured in VKM medium followed by regeneration of calli in MS13 K medium at 20°C under a 16-h photoperiod, and regenerants were multiplied on MS medium. Twenty-one somatic hybrids were
confirmed by RAPD, SSR and cytoplasm (chloroplast/mitochondria) type analysis possessing species-specific diagnostic bands
of corresponding parents. Tetraploid nature of these somatic hybrids was determined through flow cytometry analysis. Somatic
hybrids showed intermediate phenotypes (plant, leaves and floral morphology) to their parents in glass-house grown plants.
All the somatic hybrids were male-fertile. ELISA assay of somatic hybrids after artificial inoculation of Potato virus Y (PVY)
infection reveals high PVY resistance. 相似文献
18.
This paper discusses a number of experiments performed, involving the fusion by an electric field of mesophyll protoplasts
from Solanum tuberosum cv. Bintje, S. tuberosum dihaploid clones 243, 299 and the wild tuberous disease-resistant species S. bulbocastanum and S. pinnatisectum. Three fusion experiments (S. bulbocastanum + S. tuberosum dihaploid 243, S. pinnatisectum + S. tuberosum cv. Bintje and S. pinnatisectum + S. tuberosum dihaploid 299) yielded 542 calli, the 52 ones of which produced shoots. Obtained regenerants were estimated by the flow-cytometry (FC)
and RAPD analysis to determine hybrid plants.The utilisation of the FC as a useful method for detecting somatic hybrids is
also discussed in this paper. The combination S. bulbocastanum + S. tuberosum dihaploid 243 led to the creation of eight somatic hybrids, the combination S. pinnatisectum + S. tuberosum cv. Bintje yielded four somatic hybrids and the combination S. pinnatisectum + S. tuberosum dihaploid 299 resulted in no hybrid regenerants. Morphology in vitro, growth vigour and production of tuber-like structures
were evaluated in hybrid plants. Plants were transferred in vivo for further estimation (acclimatization, habitus evaluation
and tuberization ability). 相似文献
19.
Jin-Ling Dai Xiao Tan Ya-Guang Zhan Yun-Qiang Zhang Shuang Xiao Ying Gao Dong-Wei Xu Tao Wang Xiao-Chun Wang Xiang-Ling You 《Plant Cell, Tissue and Organ Culture》2011,104(1):125-130
In this work, we established a rapid and repetitive plant regeneration system for Aralia elata Seem. via primary and secondary somatic embryogenesis. Primary somatic embryogenesis was induced using leaf disks, petiole,
and root segments, individually cultured for 5 weeks on Schenk and Hildebrandt (SH) (1972) medium with 0–5.0 mg/l indolebutyric acid (IBA). Our investigation demonstrated that optimal IBA concentrations of 3.0,
2.0, and 0.3 mg/l resulted in 100% somatic embryogenesis rates and averages of 11.3, 10.0, and 8.6 somatic embryos per explant
for leaf disks, petiole, and root segments, respectively. The primary somatic embryos were used to conduct secondary somatic
embryogenesis and the following treatments, in a gradient series, were examined: 0.3–4.0 mg/l IBA, 10–70 g/l sucrose and 0.2–3.0 mg/l
abscisic acid (ABA). The results indicated that IBA was more effective than sucrose and ABA, and 3.0 mg/l IBA was the most
suitable concentration for secondary somatic embryogenesis. Histological preparations indicated a multi-cellular origin of
secondary somatic embryos and different morphological developmental stages during secondary somatic embryogenesis. Primary
and secondary somatic embryos germinated readily and developed into normal plantlets after 2 weeks in woody plant medium (WPM,
Lloyd and McCown 1980) with 20 g/l sucrose. At 4–5 cm in length, plantlets were transferred to soil (1:1 v/v of peat moss and sand) and the survival
rate was 89% after 4 weeks under greenhouse conditions. This system provides a viable contribution to A. elata gene transformation, breeding and regeneration. 相似文献
20.
Yuqiang Sun Shoumei Liu Yu Wang Brian Joseph Jones Huizhong Wang Shuijin Zhu 《Plant Cell, Tissue and Organ Culture》2011,106(3):425-433
Somatic hybrids were produced through protoplast electrofusion between the tetraploid cotton Gossypium hirsutum L. cv. ZDM-3 and the wild diploid cotton G. klotzschianum. Hybrid plants were generated from 3 out of 24 callus lines that were derived from fused protoplasts. Hybrid plants were
initially identified as somatic hybrids by ploidy analysis: the plants from the 3 callus lines had chromosome numbers near
to sum of the two parents (78 = 52 + 26). The plants from the 3 lines were subsequently confirmed as hybrids by cytological,
molecular, histological and morphological analyses. The morphology of hybrids was distinct from that of the parents, with
elongated stigmas and malformed anthers lacking microspores and pollen, leading to male sterility. It is expected that the
male sterility resulted from the high number of univalent and irregular multivalent chromosome pairings per meiocyte. 相似文献