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2.
Santy Peraza-Echeverria James L. Dale Rob M. Harding Mike K. Smith Chris Collet 《Molecular breeding : new strategies in plant improvement》2008,22(4):565-579
Most plant disease resistance (R) genes encode proteins with a nucleotide binding site and leucine-rich repeat structure (NBS-LRR).
In this study, degenerate primers were used to amplify genomic NBS-type sequences from wild banana (Musa acuminata ssp. malaccensis) plants resistant to the fungal pathogen Fusarium oxysporum formae specialis (f. sp.) cubense (FOC) race 4. Five different classes of NBS-type sequences were identified and designated as resistance gene candidates (RGCs).
The deduced amino acid sequences of the RGCs revealed the presence of motifs characteristic of the majority of known plant
NBS-LRR resistance genes. Structural and phylogenetic analyses grouped the banana RGCs within the non-TIR (homology to Toll/interleukin-1
receptors) subclass of NBS sequences. Southern hybridization showed that each banana RGC is present in low copy number. The
expression of the RGCs was assessed by RT-PCR in leaf and root tissues of plants resistant or susceptible to FOC race 4. RGC1,
3 and 5 showed a constitutive expression profile in both resistant and susceptible plants whereas no expression was detected
for RGC4. Interestingly, RGC2 expression was found to be associated only to FOC race 4 resistant lines. This finding could
assist in the identification of a FOC race 4 resistance gene. 相似文献
3.
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
4.
A high efficient protocol of Agrobacterium-mediated transformation of Musa acuminata cv. Mas (AA), a major banana variety of the South East Asia region, was developed in this study. Male-flower-derived embryogenic
cell suspensions (ECS) were co-cultivated in liquid medium with Agrobacterium strain EHA105 harboring a binary vector pCAMBIA2301 carrying nptII and gusA gene in the T-DNA. Depending upon conditions and duration of co-cultivation in liquid medium, 0–490 transgenic plants per
0.5 ml packed cell volume (PCV) of ECS were obtained. The optimum duration of inoculation was 2 h, and the highest transformation
frequency was achieved when infected ECS were co-cultivated in liquid medium first for 12 h at 40 rpm and then for 156 h at
100 rpm on a rotary shaker. Co-cultivation for a shorter duration (72 h) or shaking constantly at 100 rpm at the same duration
gave 1.6 and 1.8 folds lower transformation efficiency, respectively. No transgenic plants were obtained in parallel experiments
carried on semi-solid media. Histochemical GUS assay and molecular analysis in several tissues of the transgenic plants demonstrated
that foreign genes were stably integrated into the banana genome. Compared to semi-solid co-cultivation transformation in
other banana species, it is remarkable that liquid co-cultivation was much more efficient for transformation of the Mas cultivar,
and was at least 1 month faster for regenerating transgenic plants. 相似文献
5.
Inflorescence apices are suitable explants for the rapid in vitro propagation of Musa spp. However, the diploid and triploid banana cultivars showed different in vitro responses with respect to the hormone combinations
in Murashige and Skoog medium. The diploid cultivar (Sannachenkadali, AA) induced a maximum number of multiple shoots in 8.9 μM
6-benzyl adenine (BA) whereas the triploid cultivar (Red banana, AAA) exhibited maximum multiplication in 22.2 μM 6-benzyl
adenine. MS medium supplemented with 11.4 μM indole acetic acid and 17.8 μM BA was also suitable for shoot proliferation in
triploid cultivar but not in the diploid cultivar. The regenerated shoots were rooted in Murashige and Skoog basal medium
within 10–15 days. The rooted plantlets were transferred to vermiculite and maintained at a temperature of 25 ± 2°C for 10 days
and then at room temperature (30–32°C) for 2 weeks before transferring to potted soil compost mixture. The plantlets showed
100% survival. 相似文献
6.
Xue-Mei Dai Wang Xiao Xia Huang Jie-Tang Zhao Yun-Feng Chen Xue-Lin Huang 《In vitro cellular & developmental biology. Plant》2010,46(5):403-410
The ‘Da Jiao’ cultivar of banana (Musa paradisiacal ABB Linn.) is an ideal germplasm to produce new banana varieties resistant to Fusarium oxysporum f. sp. cubense (FOC) race 4, for this cultivar is not only a popular dessert banana in south China, but also bears high resistance to FOC
race 4. In this study, we established a homogeneous embryogenic cell suspension (ECS) of ‘Da Jiao’ and obtained regenerated
plants from ECS-derived protoplasts via somatic embryogenesis. The ECS was initiated from yellow friable callus induced from
immature male inflorescence on M1 medium. A pre-culture was used to select ECS in M2 medium without 2,4-dichlorophenoxyacetic
acid for 10 d. Addition of 1.0 mg L−1 abscisic acid to M3 medium could enhance the frequency of somatic embryogenesis by about 2.6-fold. Protoplasts, with a yield
range of 5–6 × 106 per milliliter, were isolated from the ECS. About 0.35% of the protoplasts formed microcallus, which contained about 100
cells, after 1 mo of feeder layer culture with ECS of Musa acuminate cv. Mas (AA) as nurse cells. Healthy plantlets (0.14%) were regenerated from the microcallus through somatic embryogenesis. 相似文献
7.
Banana streak virus (BSV) is a plant dsDNA pararetrovirus (family Caulimoviridae, genus badnavirus). Although integration is not an essential step in the BSV replication cycle, the nuclear genome of banana (Musa sp.) contains BSV endogenous pararetrovirus sequences (BSV EPRVs). Some BSV EPRVs are infectious by reconstituting a functional
viral genome. Recent studies revealed a large molecular diversity of episomal BSV viruses (i.e., nonintegrated) while others
focused on BSV EPRV sequences only. In this study, the evolutionary history of badnavirus integration in banana was inferred from phylogenetic relationships between BSV and BSV EPRVs. The relative evolution rates
and selective pressures (dN/dS ratio) were also compared between endogenous and episomal viral sequences. At least 27 recent independent integration events
occurred after the divergence of three banana species, indicating that viral integration is a recent and frequent phenomenon.
Relaxation of selective pressure on badnaviral sequences that experienced neutral evolution after integration in the plant
genome was recorded. Additionally, a significant decrease (35%) in the EPRV evolution rate was observed compared to BSV, reflecting
the difference in the evolution rate between episomal dsDNA viruses and plant genome. The comparison of our results with the
evolution rate of the Musa genome and other reverse-transcribing viruses suggests that EPRVs play an active role in episomal BSV diversity and evolution. 相似文献
8.
E. A. Popowich A. P. Firsov T. Y. Mitiouchkina V. L. Filipenya S. V. Dolgov V. N. Reshetnikov 《Plant Cell, Tissue and Organ Culture》2007,90(3):237-244
Transgenic plants of hyacinth (Hyacinthus orientalis L.) cvs. Edisson and Chine Pink have been obtained by Agrobacterium-mediated transformation. Leaf explants of the both hyacinth cultivars regenerated shoots on MS medium containing 2.2 μM BAP
and 0.3 μM NAA at a frequency of 95%. A. tumefaciens strain CBE21 carrying binary vector pBIThau35 was used for transformation. Plasmid pBIThau35 has been produced by cloning
preprothaumatin II cDNA into pBI121 instead of uidA gene. Inoculated leaf explants formed calli and shoots at high frequency on selective medium with 100 mg l−1 kanamycin. Four hyacinth transgenic lines of cv. Chine Pink and one line of cv. Edisson have been selected on medium containing 200 mg l−1 kanamycin. The insertion of thaumatin II gene into hyacinth genome has been confirmed by PCR-analysis. All transgenic plants
expressed substantial amounts of thaumatin II (between 0.06 and 0.28% of the total soluble protein). Hyacinth transgenic lines
were assayed for resistance to the pathogenic fungi Fusarium culmorum and Botrytis cinerea. There were no significant differences between nontransformed control and transgenic leaves of both cultivars. At the same
time the bulbs of the transgenic line Н7401 cv. Chine Pink showed the higher level of resistance to B. cinerea, the bulbs of the transgenic line Н7404 were more resistant to F. culmorum. In both cases the signs of the fungal disease were developed more slowly. The resistance of the bulbs cv. Edisson line to these fungi was not changed. All transgenic hyacinth plant were successfully transferred to soil for further evaluation. 相似文献
9.
<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of Perilla (<Emphasis Type="Italic">Perilla frutescens</Emphasis>) 总被引:1,自引:1,他引:0
Byoung-Kyu?Lee Seung-Hee?Yu Yul-Ho?Kim Byung-Ohg?Ahn Han-Sun?Hur Sang-Chul?Lee Zhanyuan?Zhang Jang-Yong?Lee
Agrobacterium tumefaciens-mediated transformation system for perilla (Perilla frutescens Britt) was developed. Agrobacterium strain EHA105 harboring binary vector pBK I containing bar and γ-tmt cassettes or pIG121Hm containing nptII, hpt, and gusA cassettes were used for transformation. Three different types of explant, hypocotyl, cotyledon and leaf, were evaluated for transformation and hypocotyl explants resulted in the highest transformation efficiency with an average of 3.1 and 2.2%, with pBK I and pIG121Hm, respectively. The Perilla spp. displayed genotype-response for transformation. The effective concentrations of selective agents were 2 mg l−1 phosphinothricin (PPT) and 150 mg l−1 kanamycin, respectively, for shoot induction and 1 mg l−1 PPT and 125 mg l−1 kanamycin, respectively, for shoot elongation. The transformation events were confirmed by herbicide Basta spray or histochemical GUS staining of T0 and T1 plants. The T-DNA integration and transgene inheritance were confirmed by PCR and Southern blot analysis of random samples of T0 and T1 transgenic plants. 相似文献
10.
In Arabidopsis, NPR1 (non-expressor of pathogenesis related genes 1, AtNPR1) functions downstream of salicylic acid (SA) and modulates the SA mediated systemic acquired resistance. It is also involved
in a cross talk with the jasmonate pathway that is essential for resistance against herbivores and necrotrophic pathogens.
Overexpression of AtNPR1 in transgenic plants resulted in enhanced disease resistance. Recently, tobacco transgenic plants expressing AtNPR1 were shown to be tolerant to the early instars of Spodoptera litura (Meur et al., Physiol Plant 133:765–775, 2008). In this communication, we show that the heterologous expression of AtNPR1 in tobacco has also enhanced the oxidative stress tolerance. The transgenic plants exhibited enhanced tolerance to the treatment
with methyl viologen. This tolerance was associated with the constitutive upregulation of PR1, PR2 (glucanase), PR5 (thaumatin like protein), ascorbate peroxidase (APX) and Cu2+/Zn2+ superoxide dismutase (SOD). This is the first demonstration of the novel function of heterologous expression of AtNPR1 in oxidative stress tolerance in transgenic tobacco. 相似文献
11.
Pascal Montoro Sébastien Lagier Christelle Baptiste Benjamin Marteaux Valérie Pujade-Renaud Julie Leclercq Laurence Alemanno 《Plant Cell, Tissue and Organ Culture》2008,94(1):55-63
In this article we describe the identification of endophytic bacteria belonging to three groups isolated from shoot tip cultures
of banana cv. Grand Naine in a recent study (Thomas et al. 2008) based on partial 16S rRNA gene sequence homology analysis. The first group included banana stocks that displayed obvious
colony growth on MS based tissue culture medium during the first in vitro passage. The second group constituted stocks that
were tissue index-negative for cultivable bacteria initially but turned index-positive after a few to several (4–8) in vitro
passages while the third group formed one sub-stock that turned index-positive after about 18 passages. The organisms belonged
to about 20 different genera comprising of α, β, γ-proteobacteria, Gram-positive firmicutes and actinobacteria. Visibly expressing easily cultured organisms during the first
in vitro passage included Enterobacter, Klebsiella, Ochrobactrum, Pantoea, Staphylococcus and Bacillus spp. Organisms of second group that were not detected or non-culturable originally constituted Brevundimonas, Methylobacterium, Alcaligenes, Ralstonia, Pseudomonas, Corynebacterium, Microbacterium, Staphylococcus, Oceanobacillus and Bacillus spp. while the third group that turned cultivable after extended in vitro culturing included mostly non-filamentous actinobacteria
(Brachybacterium, Brevibacterium, Kocuria and Tetrasphaera spp.). The identification results suggested that the endophytes of second and third groups were not strictly obligate or
fastidious microbes but those surviving in viable but-non-culturable (VBNC) state and displaying gradual activation to cultivable
form during continuous tissue culturing. Several of the organisms isolated are known as beneficial ones in agriculture while
some organisms have possible implications in human health. The use of tissue cultures for isolating uncommon endophytes is
discussed.
Supply of live bacterial cultures or genetic material for research purpose is subject to their revival from glycerol stocks
(as some of the organisms showed poor tolerance) and the requestor obtaining written permission from the Director General,
Indian Council of Agricultural Research, New Delhi-110001. 相似文献
12.
A Sebacinales species was recovered from a clone library made from a pooled rhizosphere sample of Nicotiana attenuata plants from 14 native populations. Axenic cultures of the related species, Piriformospora indica and Sebacina vermifera, were used to examine their effects on plant performance. Inoculation of N. attenuata seeds with either fungus species stimulated seed germination and increased growth and stalk elongation. S. vermifera inoculated plants flowered earlier, produced more flowers and matured more seed capsules than did non-inoculated plants. Jasmonate treatment during rosette-stage growth, which slows growth and elicits herbivore resistance traits, erased differences in vegetative, but not reproductive performance resulting from S. vermifera inoculation. Total nitrogen and phosphorous contents did not differ between inoculated and control plants, suggesting that the performance benefits of fungal inoculation did not result from improvements in nutritional status. Since the expression of trypsin proteinase inhibitors (TPI), defensive proteins which confer resistance to attack from Manduca sexta larvae, incur significant growth and fitness costs for the plant, we examined the effect of S. vermifera inoculation on herbivore resistance and TPI activity. After 10 days of feeding on S. vermifera-inoculated plants, larval mass was 46% higher and TPI activity was 48% lower than that on non-inoculated plants. These results suggest that Sebacina spp. may interfere with defense signaling and allow plants to increase growth rates at the expense of herbivore resistance mediated by TPIs. 相似文献
13.
The biotransformation of racemic 1-phenylethanol (30 mg) with plant cultured cells of basil (Ocimum basilicum cv. Purpurascens, 5 g wet wt) by shaking 120 rpm at 25°C for 7 days in the dark gave (R)-(+)-1-phenylethanol and acetophenone in 34 and 24% yields, respectively. The biotransformation can be applied to other 1-arylethanols
and basil cells oxidized the (S)-alcohols to the corresponding ketones remaining the (R)-alcohols in excellent ee. 相似文献
14.
Wang Xiao Xia Huang Qing Gong Xue-Mei Dai Jie-Tang Zhao Yue-Rong Wei Xue-Lin Huang 《Plant Cell, Tissue and Organ Culture》2009,97(3):313-321
To attempt to introduce genetic information of disease resistance from Musa acuminata cv. Mas (AA) to Musa silk cv. Guoshanxiang (AAB) and obtain somatic hybrids, we developed an asymmetric protoplast fusion with 20% (w/v) polyethylene
glycol (PEG). The protoplasts derived from embryogenic suspension cultural cells of cv. Guoshanxiang (AAB) and cv. Mas (AA)
were, respectively treated with 1.5 mM iodoacetamide (IOA) and with ultraviolet light (UV) at an intensity of 50 W/m2 for 120 s. A total of 47 regenerated green plants were obtained and eight of which were survived in greenhouse. Six of the
survived plants were identified as hybrids by RAPD analysis and only three hybrids were retained vigorously in field. The
hybrid nature of the three plants was further confirmed according to their ISSR (inter-simple sequence repeat) patterns and
the results indicated that they were true somatic hybrids. Chromosome analysis revealed that the three hybrids possessed an
aneuploid chromosome number (2n = 34). 相似文献
15.
A bacterial artificial chromosome (BAC) library of banana (Musa acuminata) was used to select BAC clones that carry low amounts of repetitive DNA sequences and could be suitable as probes for fluorescence
in situ hybridization (FISH) on mitotic metaphase chromosomes. Out of eighty randomly selected BAC clones, only one clone gave a
single-locus signal on chromosomes of M. acuminata cv. Calcutta 4. The clone localized on a chromosome pair that carries a cluster of 5S rRNA genes. The remaining BAC clones
gave dispersed FISH signals throughout the genome and/or failed to produce any signal. In order to avoid the excessive hybridization
of repetitive DNA sequences, we subcloned nineteen BAC clones and selected their ‘low-copy’ subclones. Out of them, one subclone
gave specific signal in secondary constriction on one chromosome pair; three subclones were localized into centromeric and
peri-centromeric regions of all chromosomes. Other subclones were either localized throughout the banana genome or their use
did not result in visible FISH signals. The nucleotide sequence analysis revealed that subclones, which localized on different
regions of all chromosomes, contained short fragments of various repetitive DNA sequences. The chromosome-specific BAC clone
identified in this work increases the number of useful cytogenetic markers for Musa. 相似文献
16.
The species Solanum surattense Burm.f. has importance in ayurvedic medicine and also as vegetable. Streptomycin-resistant plantlets were induced showing chloroplast encoded mutants in S. surattense from mutagenised (ethyl methane sulphonate and gamma-rays) cotyledon explants. Chloroplast encoded – streptomycin resistant – shoots were developed from green (unbleached) sectors of the cotyledons. The streptomycin-resistant plants were similar to parental plants in morphology and ploidy level (2n=2x=24). Reciprocal crosses between streptomycin-resistant and the original streptomycin sensitive plants have shown the non-Mendelian transmission under the control of chloroplast – DNA. These antibiotic resistant plants are useful in designing biochemical selection schemes aimed at somatic hybrid/cybrid recovery in S. surattense. 相似文献
17.
Three constructs harbouring novel Bacillus thuringiensis genes (Cry1C, Cry2A, Cry9C) and bar gene were transformed into four upland cotton cultivars, Ekangmian10, Emian22, Coker201 and YZ1 via Agrobacterium-mediated transformation. With the bar gene as a selectable marker, about 84.8 % of resistant calli have been confirmed positive by polymerase chain reaction (PCR)
tests, and totally 50 transgenic plants were regenerated. The insertions were verified by means of Southern blotting. Bioassay
showed 80 % of the transgenic plantlets generated resistance to both herbicide and insect. We optimized conditions for improving
the transformation efficiency. A modified in vitro shoot-tip grafting technique was introduced to help entire transplantation. This result showed that bar gene can replace antibiotic marker genes (ex. npt II gene) used in cotton transformation. 相似文献
18.
Phytophthora spp., some of the more important casual agents of plant diseases, are responsible for heavy economic losses worldwide. Plant
defensins have been introduced as transgenes into a range of species to increase host resistance to pathogens to which they
were originally susceptible. However, the effectiveness and mechanism of interaction of the defensins with Phytophthora spp. have not been clearly characterized in planta. In this study, we expressed the Dahlia merckii defensin, DmAMP1, in papaya (Carica papaya L.), a plant highly susceptible to a root, stem, and fruit rot disease caused by Phytophthora palmivora. Extracts of total leaf proteins from transformed plants inhibited growth of Phytophthora in vitro and discs cut from the leaves of transformed plants inhibited growth of Phytophthora in a bioassay. Results from our greenhouse inoculation experiments demonstrate that expressing the DmAMP1 gene in papaya plants increased resistance against P. palmivora and that this increased resistance was associated with reduced hyphae growth of P. palmivora at the infection sites. The inhibitory effects of DmAMP1 expression in papaya suggest this approach has good potential to impart transgenic resistance against Phytophthora in papaya. 相似文献
19.
S. Fondevilla D. Rubiales M. T. Moreno A. M. Torres 《Molecular breeding : new strategies in plant improvement》2008,22(2):193-200
Three genes, er1, er2 and Er3, conferring resistance to powdery mildew (Erysiphe pisi) in pea have been described so far. Because single gene-controlled resistance tends to be overcome by evolution of pathogen
virulence, accumulation of several resistance genes into a single cultivar should enhance the durability of the resistance.
Molecular markers linked to genes controlling resistance to E. pisi may facilitate gene pyramiding in pea breeding programs. Molecular markers linked to er1 and er2 are available. In the present study, molecular markers linked to Er3 have been obtained. A segregating F2 population derived from the cross between a breeding line carrying the Er3 gene, and the susceptible cultivar ‘Messire’ was developed and genotyped. Bulk Segregant Analysis (BSA) was used to identify
Random Amplified Polymorphic DNA (RAPD) markers linked to Er3. Four RAPD markers linked in coupling phase (OPW04_637, OPC04_640, OPF14_1103, and OPAH06_539) and two in repulsion phase
(OPAB01_874 and OPAG05_1240), were identified. Two of these, flanking Er3, were converted to Sequence Characterized Amplified Region (SCAR) markers. The SCAR marker SCW4637 co-segregated with the resistant gene, allowing the detection of all the resistant individuals. The SCAR marker SCAB1874, in repulsion phase with Er3, was located at 2.8 cM from the gene and, in combination with SCW4637, was capable to distinguish homozygous resistant individuals from heterozygous with a high efficiency. In addition, the validation
for polymorphism in different genetic backgrounds and advanced breeding material confirmed the utility of both markers in
marker-assisted selection. 相似文献
20.
Xiuli Shen Jianjun Chen Michael E. Kane Richard J. Henny 《Plant Cell, Tissue and Organ Culture》2007,91(1):21-27
The occurrence of somaclonal variation among regenerants derived through indirect shoot organogenesis from leaf explants of
three Dieffenbachia cultivars Camouflage, Camille and Star Bright was evaluated. Three types of somaclonal variants (SV1, SV2, and SV3) were
identified from regenerated plants of cv. Camouflage, one type from cv. Camille, but none from cv. Star Bright. The three
variants had novel and distinct foliar variegation patterns compared to cv. Camouflage parental plants. Additionally, SV1
was taller with a larger canopy and longer leaves than parental plants and SV2. SV2 and SV3 did not produce basal shoots (single
stem) but basal shoot numbers between SV1 and parental plants were similar ranging from three to four. The variant type identified
from regenerated cv. Camille had lanceolate leaves compared to the oblong leaves of the parent. This variant type also grew
taller and had a larger canopy than parental plants. The rates of somaclonal variation were up to 40.4% among regenerated
cv. Camouflage plants and 2.6% for regenerated cv. Camille. The duration of callus culture had no effect on somaclonal variation
rates of cv. Camouflage as the rates between plants regenerated from 8 months to 16 months of callus culture were similar.
The phenotypes of the identified variants were stable as verified by their progenies after cutting propagation. This study
demonstrated the potential for new cultivar development by selecting callus-derived somaclonal variants of Dieffenbachia. 相似文献