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1.
RFLP analysis to identify putative chromosomal regions involved in the anther culture response and callus formation of maize 总被引:4,自引:0,他引:4
Y. Wan T. R. Rocheford J. M. Widholm 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1992,85(2-3):360-365
Summary RFLP analysis was performed with anther culture-derived callus lines developed from the maize F1 hybrids Pa91 x FR16 (PF), H99 x Pa91 (HP) and H99 x FR16 (HF). Relatively evenly spaced RFLP markers were selected to cover the maize genome with 52, 58 and 35 RFLP markers for the PF, HP and HF callus lines, respectively. The results from populations PF and HP combined with limited information from HF showed that six chromosomal regions on chromosomes 1, 2 (two regions), 3, 6 and 8 appear to be associated with the formation of embryo-like structures (ELSs) from microspores or the subsequent formation of regenerable callus from the ELSs. Regions at the end of the long arm of chromosome 2 and on the long arm of chromosome 8 appear to be associated with ELS formation, and the other regions appear to be associated with either ELS or regenerable callus formation or both. Certain regions that we have identified are the same as those found in other studies to be important for friable, embryogenic callus formation (chromosomes 1 and 3 and near the centromere of 2) and for ESL formation (chromosomes 1 and 3). This study has provided evidence for the genetic basis of the maize anther culture response and callus formation. 相似文献
2.
V. M. Peschke R. L. Phillips 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1991,81(1):90-97
Summary Previous experiments have revealed that the maize transposable element Activator (Ac) may become active during tissue culture. The objective of the present study was to determine whether a second transposable element, Suppressor-mutator (Spm), could also be activated in tissue culture and detected in regenerated maize plants. Approximately 500 R1 progeny of 143 regenerated plants (derived from 49 embryo cell lines) were crossed as males onto an Spm-responsive tester stock. Spm activity was observed in two R1 progeny of a single regenerated plant. This plant had been regenerated from Type II (friable embryogenic) callus of an A188 × B73 genetic background after 8 months in culture; the absence of Spm activity in four other plants regenerated from this same callus demonstrates that Spm activity was not present before culturing. Approximately 20 Spm-homologous DNA sequences were detected in each of the inbreds used to initiate the tissue cultures; it is presumed that one of these became active to give rise to Spm activity. 相似文献
3.
Peng Jiao Ruiqi Ma Zhuo Qi Zhenzhong Jiang Siyan Liu Jing Qu Shuyan Guan Yiyong Ma 《Phyton》2020,89(1):121-130
This research uses the immature embryos of inbred maize lines
(GSH9901, Hi01, Hi02, and Chang 7-2) as receptor materials to establish the callus
induction system. These inbred lines provide the receptor materials for the genetic
regeneration of maize and the verification of the genetic functions of maize. The
factor experiment and orthogonal experiments were used to investigate the impacts
of different genotypes, immature embryo size, shield orientation, 2, 4-D
concentration, proline concentration, and folic acid concentration on the induction
rate of embryogenic callus tissue. A sensitivity experiment testing glyphosate (Bar)
and an antibiotic (Cefotaxime sodium) were also conducted. The results indicate
that the immature embryos of inbred maize line GSH9901 were the most effective
for callus tissue induction, and the immature embryos with a length of 1.6-2.0 mm
produce the best result. The upward shield face is more successful for the
formation of induced callus. Using orthogonal analysis, we found that the optimal
combination for the induction system was A3 (2,4-D concentration 0.25 mg mL-1
),
B1C3 (proline concentration 0.8 mg mL-1
), and D2 (folate Concentration 0.5 mg
mL-1) and the induction rate reached 84%. We found that cold storage at 4 °C for 1
d is more conducive for the formation of embryogenic callus than the other
treatments tested. The sensitivity experiment for callus tissue screening revealed
the critical concentration of glyphosate to be 10 mg ml-1
, and the critical
concentration of antibiotic is 250 mg ml-1
. Using this combination of glyphosate
and antibiotic resulted in regenerated plants. This study established the optimal
conditions for immature embryo callus tissue induction in maize. 相似文献
4.
Androgenic lines of Brassica juncea cv. PR-45 raised by anther culture, were screen for genetic variation. 393 androgenic plants were transferred to pots to study the R0 generation. These plants showed substantial variation for different characters. Seed progenies of 27 lines of R0 plants were sown in the field to study R1 generation. Androgenic plants within lines were significantly homogeneous for the various agronomic characters studied. Two lines were shorter (18 – 20 %) than the control plants, with a remarkable feature of early maturation. Three lines showed 27 – 31 % higher yield than the parent cultivar. 相似文献
5.
Fertile plants have been obtained from maize (Zea mays L.) embryogenic suspension culture protoplasts. Friable, embryogenic callus initiated from an immature embryo from a cross involving the genotypes A188, B73, and Black Mexican sweetcorn was used to establish a rapidly growing embryogenic suspension culture. After nine months in culture, high yields of viable protoplasts (30×106/ gram fresh weight) were obtained following a 1.5 hour enzymatic digestion. Protoplasts cultured with feeder cells divided and formed embryogenic callus, from which male and female fertile plants were regenerated. Protoplast-derived R1 plants were self-pollinated and immature R2 embryos isolated for callus initiation. Female fertile plants have also been produced from protoplasts isolated from an R2-derived suspension culture. Significant interactions between protoplast and feeder-cell lines were observed.Abbreviations BC
backcross
- BMS
Black Mexican Sweetcorn
- 2,4-D
2,4-dichlorophenoxyacetic acid
- PWS
protoplast wash solution (0.2 M mannitol, 80 mM CaCl2)
- FDA
fluorescein diacetate
- ABA
abscisic acid 相似文献
6.
To develop a less genotype-dependent maize-transformation procedure, we used 10-month-old Type I callus as target tissue for microprojectile bombardment. Twelve transgenic callus lines were obtained from two of the three anther-culture-derived callus cultures representing different gentic backgrounds. Multiple fertile transgenic plants (T0) were regenerated from each transgenic callus line. Transgenic leaves treated with the herbicide Basta showed no symptoms, indicating that one of the two introduced genes, bar, was functionally expressing. Data from DNA hybridization analysis confirmed that the introduced genes (bar and uidA) were integrated into the plant genome and that all lines derived from independent transformation events. Transmission of the introduced genes and the functional expression of bar in T1 progeny was also confirmed. Germination of T1 immature embryos in the presence of bialaphos was used as a screen for functional expression of bar; however, leaf painting of T1 plants proved a more accurate predictor of bar expression in plants. This study suggests that maize Type I callus can be transformed efficiently through microprojectile bombardment and that fertile transgenic plants can be recovered. This system should facilitate the direct introduction of agronomically important genes in to commercial genotypes. 相似文献
7.
This study was conducted to determine the reciprocal effects for anther culture response in wheat (Triticum aestivum L.) using a set of 4 × 4 full diallel crosses. Both reciprocal and nuclear genetic effects were highly significant for anther
culture response and useful for selection and breeding purposes. General combining ability (GCA) effects were predominant
for all investigated anther culture traits. Also, significant differences for specific combining ability (SCA) effects were
detected between reciprocal crosses. Although significant reciprocal differences for responding anther, callus number and
green plant regeneration were recorded in some reciprocal crosses, there were no significant reciprocal differences for albino
plant regeneration. The use of one parent as male or female could lead to change at the production of green plants from the
F1 hybrids and screening of inbred lines for response to anther culture, without reciprocal effects, could decrease the utilization
of breeding material. 相似文献
8.
Massiah Andrea Rong Hong Lin Brown Sherralyn Laurie Sophie 《Molecular breeding : new strategies in plant improvement》2001,7(2):163-173
Anther culture has been developed in the winter wheat cultivar Florida to achieve accelerated production and identification of homozygous transgenic lines. With untransformed, seed-derived plants to develop the culture system, it was shown that cold pre-treatment of spikes excised from donor plants and addition of 2,4-dichlorophenoxyacetic acid together with either kinetin or 6-benzylaminopurine in the callus induction medium improves the anther culture response. The procedure developed allowed production of fertile homozygous lines within 8–9 months, which includes an 8-week vernalisation period. With transgenic wheat plants produced by particle bombardment as donors, we show that the system can be used to produce homozygous transgenics, requiring one generation cycle. Both T0 tissue culture-derived plants and their T1 seed-derived descendents serve as suitable donors. We show that an anther culture response comparable to that of untransformed, seed-derived plants can be achieved with T0 tissue culture-derived plants. PCR and Southern molecular analyses of anther culture-derived transgenics show that the transgenes are stably inherited; there are no perturbations at the chromosomal level around the sites of transgene integration as a result of in vitro chromosome manipulation during anther culture. 相似文献
9.
Feiyu Tang Yazhong Tao Tianyong Zhao Guoying Wang 《Plant Cell, Tissue and Organ Culture》2006,84(2):100210-100214
Haploid induction has potential application for maize breeding. This paper reports that maize haploid plants have been induced
by in vitro culture of pollinated ovaries. From a total of 26,400 cultured ovaries, 24 haploid plants were obtained and two of them were
doubled after colchicine treatment. The maximum frequency of gynogenesis was 0.17% at 19.5 h post-pollination (HPP). The results
showed that HPP was an important factor affecting plant induction from ovaries. Regenerated diploid R0 plants were then subjected to genetic analysis using SSR molecular markers. One R0 plant, whose progeny revealed a high level of homogeneity for several agro-morphological traits, was homozygous at 20 loci
tested, with 11 showing paternal and 9 maternal banding pattern. This demonstrates that it is feasible to induce maize haploid
plants by in vitro culture of pollinated ovaries. 相似文献
10.
Summary Plants derived from anther culture are theoretically haploid, but diploid plants are also known to arise. Anther culture-derived
diploid plants are usually homozygous and are believed to be due to spontaneous doubling of chromosomes in either microsporocytes
or callus cells during the culture process. However, heterozygous diploid regenerants may also arise from a) regeneration
from cultured somatic cells, b) mutation occurring during or after a spontaneous doubling event, c) fusion of unlike haploid
cells in chimeric callus, and d) regeneration from diploid microsporocytes resulting from aberrant meioses. This study was
designed to elucidate the frequency and origin of diploid regenerants from rice anther culture. Regenerants were obtained
from 11 F1 genotypes. Progeny testing detected heterozygosity in 7 out of 211 regenerants. Each of the heterozygous regenerants were
from ‘Calrose 76’/waxy ‘M-101’, Half of the diploid regenerants from this cross were heterozygous. No heterozygous regenerants
arose from the other 10 F1 genotypes. Progeny testing indicated that two of the heterozygous regenerants were as heterozygous as the F1 plants for three parental characters. The other five regenerants exhibited decreased levels of heterozygosity. One of the
heterozygous regenerants exhibited evidence of mutation for a non-parental character. However, mutation is an unlikely cause
of the observed high levels of parental-type heterozygosity. No evidence for the occurrence of chimeric callus was detected,
making this an unlikely cause as well. The most likely origin of the observed partial heterozygosity is regeneration from
diploid microspores, which could also produce plants exhibiting complete parental-type heterozygosity. 相似文献
11.
Selection for increased anther culture response in maize 总被引:1,自引:0,他引:1
J. F. Petolino A. M. Jones S. A. Thompson 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1988,76(1):157-159
Summary Anther culture of a three-way cross, (H99 × FR16) × Pa91, resulted in the regeneration of two anther-derived plants which were crossed to produce an F1 progeny. Fourteen S1 families derived from this cross were evaluated for their anther culturability. Dramatic increases in the level of androgenesis, expressed as the percentage of cultured anthers which produced embryo-like structures, were observed. An overall mean response frequency of 23.4% was observed for the S1 families. This was compared to a 3.5% response in the original three-way cross. These results demonstrate that genetic improvement of in vitro androgenesis in maize is possible and that anther culture per se constitutes a procedure for selecting genes which favor increased levels of response. 相似文献
12.
Expression of a synthetic E. coli heat-labile enterotoxin B sub-unit (LT-B) in maize 总被引:2,自引:0,他引:2
Chikwamba Rachel McMurray Jennifer Shou Huixia Frame Bronwyn Pegg Sue Ellen Scott Paul Mason Hugh Wang Kan 《Molecular breeding : new strategies in plant improvement》2002,10(4):253-265
We have produced the B subunit of the enterotoxigenic Escherichia coli (ETEC) heat-labile enterotoxin (LT-B) in transgenic maize seed. LT-B is a model antigen that induces a strong immune response upon oral administration and enhances immune responses to conjugated and co-administered antigens. Using a synthetic LT-B gene with optimized codon sequence, we examined the role of promoters and the SEKDEL endoplasmic reticulum retention motif in LT-B accumulation in callus and in kernels. Two promoters, the constitutive CaMV 35S promoter and the maize 27 kDa gamma zein promoter, which directs endosperm-specific gene expression in maize kernels, regulated LT-B expression. Ganglioside-dependent ELISA analysis showed that using the constitutive promoter, maximum LT-B level detected in callus was 0.04% LT-B in total aqueous-extractable protein (TAEP) and 0.01% in R1 kernels of transgenic plants. Using the gamma zein promoter, LT-B accumulation reached 0.07% in R1 kernels. The SEKDEL resulted in increased LT-B levels when combined with the gamma zein promoter. We monitored LT-B levels under greenhouse and field conditions over three generations. Significant variability in gene expression was observed between transgenic events, and between plants within the same event. A maximum of 0.3% LT-B in TAEP was measured in R3 seed of a transgenic line carrying CaMV 35S promoter/LT-B construct. In R3 seed of a transgenic line carrying the gamma zein promoter/LT-B construct, up to 3.7% LT-B in TAEP could be detected. We concluded that maize seed can be used as a production system for functional antigens. 相似文献
13.
Type II callus cultures were initiated from immature tassels of a maize genotype with an A188/B73 genetic background using N6 medium containing 1.0 mg/liter 2,4-D, 100 mg/liter casamino acids, 25 mM proline, and 0.2% phytagel™. Inclusion of 10 μM AgNO3 in this medium significantly increased the frequency and vigor of the type II callus response. Friable calli emerged from these explants after two consecutive 2-week subculture intervals. Tassels from 10 to 30 mm long were capable of producing type II cultures. The plants regenerated from these cultures were green and indistinguishable from plants regenerated from immature embryo-derived calli. 相似文献
14.
J. C. Afele L. W. Kannenberg 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1990,80(4):459-464
Summary Anthers of two maize (Zea mays L.) inbred lines, DBTS (P1) and B73 (P2), their F1, F2 and first backcross generations — F1 x DBTS (B1), and F1 x B73 (B2) — were float cultured in YP medium to study the inheritance of corn anther culturability using generation mean analysis. Significant effects of generation were observed for the three traits measured: anther response (%), frequency of embryos (%) and anther productivity. Variation among the generations was similar for anther response and frequency of embryos: no significant differences were found among the P1, F1, F2 and B1 means, but the means of P2 and B2 were significantly lower than those of the other generations. For anther productivity, the F2 generation tended to have a slightly higher tendency for multiple embryo formation. A simple additive-dominance model was adequate in explaining the inheritance of anther response and frequency of embryos, but digenic epistasis (additive x dominance) was involved in the inheritance of anther productivity. Additive genetic variance was higher than non-additive genetic variance for all the traits; however, only environmental variance was significant. Narrow-sense heritability estimates were 65% and 75% for anther response and frequency of embryos, respectively. Significant inter-plant variation was observed within generations, even for the inbred line DBTS, but isozymic analysis involving five enzyme loci did not reveal any genotypic variability within the inbred lines DBTS and B73. 相似文献
15.
Efficient production of doubled haploid plants through colchicine treatment of anther-derived maize callus 总被引:3,自引:0,他引:3
Y. Wan J. F. Petolino J. M. Widholm 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1989,77(6):889-892
Summary A chromosome doubling technique, involving colchicine treatment of an embryogenic, haploid callus line of maize (Zea mays L., derived through anther culture), was evaluated. Two colchicine levels (0.025% and 0.05%) and three treatment durations (24, 48, and 72 h) were used and compared to untreated controls. Chromosome counts and seed recovery from regenerated plants were determined. No doubled haploid plants were regenerated from calli without colchicine treatment. After treatment with colchicine for 24 h, the callus tissue regenerated about 50% doubled haploid plants. All of the plants regenerated from the calli treated with colchicine for 72 h were doubled haploids, except for a few tetraploid plants. No significant difference in chromosome doubling was observed between the two colchicine levels. Most of the doubled haploid plants produced viable pollen and a total of 107 of 136 doubled haploid plants produced from 1 to 256 seeds. Less extensive studies with two other genotypes gave similar results. These results demonstrate that colchicine treatment of haploid callus tissue can be a very effective and relatively easy method of obtaining a high frequency of doubled haploid plants through anther culture. 相似文献
16.
Production of transgenic maize plants and progeny by bombardment of hi-II immature embryos 总被引:4,自引:0,他引:4
D. D. Songstad C. L. Armstrong W. L. Petersen B. Hairston M. A. W. Hinchee 《In vitro cellular & developmental biology. Plant》1996,32(3):179-183
Summary Production of transgenic maize (Zea mays L.) callus, plants, and progeny from microprojectile bombardment of 2–5-d cultured Hi-II immature embryos is described. Histological
evidence indicates that these tissues are amenable to transformation due to surface layer cell division of the scutellum.
Two out of every 100 bombarded embryos produced transgenic callus and R0 transgenic plants were both male and female fertile. Expected segregation of transgenes was observed in progeny. The primary
advantage of bombarding these tissues is increased male and female fertility of transgenic plants compared with those produced
using long-term callus or suspension cultures. 相似文献
17.
H. Ma R. H. Busch O. Riera-Lizarazu H. W. Rines R. Dill-Macky 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,99(3-4):432-436
Anther culture and maize hybridization are two frequently used techniques for doubled haploid production in wheat (Triticum aestivum L.). Information on the field performance of lines derived from these techniques is limited. This study was conducted to
compare the performance of F4:6 lines obtained by single-seed descent with lines obtained by anther culture and maize (Zea mays L.) pollination from the same cross of spring wheat, ’Chris’/MN 7529. Thirty-three lines derived from each of those techniques
were evaluated in six environments for grain yield, protein content, test weight, heading date, kernel weight and plant height.
Mean performance of the single-seed descent lines exceeded performance of the anther culture lines for grain yield, kernel
weight and plant height with no apparent differences for grain protein content, test weight and heading date. No differences
between trait means for the single-seed descent and maize pollination lines were found except for plant height. The best 5
lines from each method for grain yield, protein content and test weight were similar in performance except that the protein
content was higher for the maize pollination lines than for the single-seed descent lines. Acceptable levels of agronomic
performance could be found among lines from each method. Wide acceptance of the doubled haploid technique for pure line production
in breeding programs may, however, be limited by the often poor efficiency of doubled haploid line production, resulting in
smaller population sizes for selection of desirable traits in comparison to the single-seed descent method.
Received: 31 July 1998 / Accepted: 28 November 1998 相似文献
18.
Anthers of two rice (Oryza sativa L.) varieties, Lunhui 422 (P1) and Jinzao 5 (P2), their F1, F2 and first backcross generation-F1 x Lunhui 422 (B1), and F1 x Jinzao 5 (B2)-were cultured in L8 medium to study the inheritance of rice anther culturability using generation mean analysis. Significant effects of generation were observed for the four traits measured: anther response (%), frequency of callus induction (%), frequency of callus differentiation (%) and culture efficiency (%). Variation among the generations was similar for all traits: significant differences were found among six generations and the means of the P2 and B2 were significantly lower than those of the other generations. The frequency of callus differentiation showed a nonsignificant difference among the P1, F1, F2 and B1 generations which had slightly high values than the P2 and B2. Additive genetic variance (VA) was higher than non-additive genetic variance (VD) for anther response and frequency of callus induction. However, AV was lower than VD for frequence of callus differentiation and culture efficiency, VA was significant for all traits except for the culture efficiency, and VD was nonsignificant for all traits except for the frequency of callus differentiation. On the other hand, environmental variation (VE) was significant for the 4 traits. Narrow-sense heritability estimates were 95.52%, 82.19% and 13.54% for anther response, frequency of callus induction and culture efficiency, respectively.Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid 相似文献
19.
Developmental variability was introduced into Withania somnifera using genetic transformation by Agrobacterium rhizogenes, with the aim of changing withasteroid production. Inoculation of W. somnifera with A. rhizogenes strains LBA 9402 and A4 produced typical transformed root lines, transformed callus lines, and rooty callus lines with simultaneous
root dedifferentiation and redifferentiation. These morphologically distinct transformed lines varied in T-DNA content, growth
rates, and withasteroid accumulation. All of the lines with the typical transformed root morphology contained the TL T-DNA, and 90% of them carried the TR T-DNA, irrespective of the strain used for infection. Accumulation of withaferin A was maximum (0.44% dry weight) in the
transformed root line WSKHRL-1. This is the first detection of withaferin A in the roots of W. somnifera. All of the rooty callus lines induced by strain A4 contained both the TL and the TR-DNAs. In contrast, 50% of the rooty-callus lines obtained with strain LBA 9402 contained only the TR T-DNA. All the rooty callus lines accumulated both withaferin A and withanolide D. The callusing lines induced by LBA 9402
lacked the TL T-DNA genes, while all the callusing lines induced by strain A4 contained the TL DNA. Four of these callus lines produced both withaferin A (0.15–0.21% dry weight) and withanolide D (0.08–0.11% dry weight),
and they grew faster than the transformed root lines. This is the first report of the presence of withasteroids in undifferentiated
callus cultures of W. somnifera. 相似文献
20.
Improved Agrobacterium-mediated transformation of three maize inbred lines using MS salts 总被引:3,自引:0,他引:3
Frame BR McMurray JM Fonger TM Main ML Taylor KW Torney FJ Paz MM Wang K 《Plant cell reports》2006,25(10):1024-1034
Transformation technology as a research or breeding tool to improve maize is routinely used in most industrial and some specialized public laboratories. However, transformation of many inbred lines remains a challenging task, especially when using Agrobacterium tumefaciens as the delivery method. Here we report success in generating transgenic plants and progeny from three maize inbred lines using an Agrobacterium-mediated standard binary vector system to target maize immature embryos. Eleven maize inbred lines were pre-screened for transformation frequency using N6 salts. A subset of three maize inbred lines was then systematically evaluated for frequency of post-infection embryogenic callus induction and transformation on four media regimes: N6 or MS salts in each of two distinct media backgrounds. Transgenic plants recovered from inbred lines B104, B114, and Ky21 were analyzed for transgene integration, expression, and transmission. Average transformation frequencies of 6.4% (for B104), 2.8% (for B114), and 8% (for Ky21) were achieved using MS salts. Availability of Agrobacterium-mediated maize inbred line transformation will improve future opportunities for maize genetic and functional genomic studies. 相似文献