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1.
A disease resistance related gene, MbR7, was identified in the wild apple species, Malus baccata. The MbR7 gene has a single open reading frame (ORF) of 3,288 nucleotides potentially encoding a 1,095-amino acid protein. Its deduced amino acid sequence resembles the N protein of tobacco and the NL27 gene of potato and has several motifs characteristic of a TIR-NBS-LRR R gene subclass. Ectopic expression of MbR7 in Arabidopsis enhanced the resistance against a virulent pathogen, Pseudomonas syringae pv. tomato DC3000. Microarray analysis confirmed the induction of defense-related gene expression in 35S::MbR7 heterologous Arabidopsis plants, indicating that the MbR7 gene likely activates a downstream resistance pathway without interaction with pathogens. Our results suggest that MbR7 can be a potential target gene in developing a new disease-resistant apple variety. 相似文献
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V. G. M. Bus D. Chagné H. C. M. Bassett D. Bowatte F. Calenge J.-M. Celton C.-E. Durel M. T. Malone A. Patocchi A. C. Ranatunga E. H. A. Rikkerink D. S. Tustin J. Zhou S. E. Gardiner 《Tree Genetics & Genomes》2008,4(2):223-236
Woolly apple aphid (WAA; Eriosoma lanigerum Hausm.) can be a major economic problem to apple growers in most parts of the world, and resistance breeding provides a sustainable
means to control this pest. We report molecular markers for three genes conferring WAA resistance and placing them on two
linkage groups (LG) on the genetic map of apple. The Er1 and Er2 genes derived from ‘Northern Spy’ and ‘Robusta 5,’ respectively, are the two major genes that breeders have used to date
to improve the resistance of apple rootstocks to this pest. The gene Er3, from ‘Aotea 1’ (an accession classified as Malus sieboldii), is a new major gene for WAA resistance. Genetic markers linked to the Er1 and Er3 genes were identified by screening random amplification of polymorphic deoxyribonucleic acid (DNA; RAPD) markers across DNA
bulks from resistant and susceptible plants from populations segregating for these genes. The closest RAPD markers were converted
into sequence-characterized amplified region markers and the genome location of these two genes was assigned to LG 08 by aligning
the maps around the genes with a reference map of ‘Discovery’ using microsatellite markers. The Er2 gene was located on LG 17 of ‘Robusta 5’ using a genetic map developed in a M.9 × ‘Robusta 5’ progeny. Markers for each of
the genes were validated for their usefulness for marker-assisted selection in separate populations. The potential use of
the genetic markers for these genes in the breeding of apple cultivars with durable resistance to WAA is discussed. 相似文献
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Multiple virus resistance in transgenic plants conferred by the human dsRNA-dependent protein kinase
Pyung Ok Lim Ung Lee Jong Sang Ryu Jang Kyung Choi Ara Hovanessian Cheol Soo Kim Baik Ho Cho Hong Gil Nam 《Molecular breeding : new strategies in plant improvement》2002,10(1-2):11-18
We have developed a new strategy for engineering resistance to multipleviruses in plants. The strategy exploits the human double stranded (ds)RNA-dependent protein kinase (PKR). PKR is one of theinterferon-induced enzymes. It confers viral resistance in mammals byinhibitingviral replication through the inactivation of the translational initiationfactor, eIF-2, upon activation by dsRNA. The humanPKR gene was fused to the promoter of theArabidopsis blue copper binding protein gene(BCB) that is induced rapidly in response to wounding. Thechimeric gene cassette was introduced into tobacco plants. Expression of thePKR gene in transgenic tobacco plants was demonstrated byRNA gel blot analysis and autophosphorylation assay of anM
r 68,000 protein. The transgenic plantsexpressing the PKR gene showed significantly reduced viralsymptoms or no viral symptoms at all, when challenged by different plant RNAviruses, such as Cucumber mosaic virus, Tobaccoetch virus, or Potato virus Y. Thus, expressionof a single component in the human interferon pathway, thePKR gene, can effectively confer resistance to multipleviruses in transgenic plants. 相似文献
7.
K. D. Jermstad L. A. Sheppard B. B. Kinloch A. Delfino-Mix E. S. Ersoz K. V. Krutovsky D. B. Neale 《Tree Genetics & Genomes》2006,2(2):76-85
The nucleotide-binding-site and leucine-rich-repeat (NBS–LRR) class of R proteins is abundant and widely distributed in plants. By using degenerate primers designed on the NBS domain in lettuce, we amplified sequences in sugar pine that shared sequence identity with many of the NBS–LRR class resistance genes catalogued in GenBank. The polymerase chain reaction products were used to probe a cDNA library constructed from needle tissue of sugar pine seedlings. A full-length cDNA was obtained that demonstrated high predicted amino acid sequence similarity to the coiled coil (CC)–NBS–LRR subclass of NBS–LRR resistance proteins in GenBank. Sequence analyses of this gene in megagametophytes from two sugar pine trees segregating for the hypersensitive response to white pine blister rust revealed zero nucleotide variation. Moreover, there was no variation found in 24 unrelated sugar pine trees except for three single-nucleotide polymorphisms located in the 3′ untranslated region. Compared to other genes sequenced in Pinaceae, such a low level of sequence variation in unrelated individuals is unusual. Although, numerous studies have reported that plant R genes are under diversifying selection for specificity to evolving pathogens, the resistance gene analog discussed here appears to be under intense purifying selection.An erratum to this article can be found at 相似文献
8.
Insecticidal proteins are a potential resource to enhance resistance to insect pests in transgenic plants. Here, we describe
the generation and analysis of the apple cultivar ‘Royal Gala’ transgenic for Nicotiana alata (N. alata) proteinase inhibitor (PI) and the impact of this PI on the growth and development of the Epiphyas postvittiana (light-brown apple moth). A cDNA clone encoding a proteinase inhibitor precursor from N. alata (Na-PI) under the control of either a double 35S promoter or a promoter from a ribulose-1,5-bisphosphate carboxylase small
sub-unit gene (rbcS-E9 promoter) was stably incorporated into ‘Royal Gala’ apple using Agrobacterium-mediated transformation. A 40.3 kDa Na-PI precursor protein was expressed and correctly processed into 6-kDa proteinase inhibitors
in the leaves of transgenic apple lines. The 6-kDa polypeptides accumulated to levels of 0.05 and 0.1% of the total soluble
protein under the control of the rbc-E9 promoter and the double 35S promoter, respectively. Light-brown apple moth larvae fed with apple leaves expressing Na-PI
had significantly reduced body weight after 7 days of feeding and female pupae were 19–28% smaller than controls. In addition,
morphological changes such as pupal cases attached to the wing, deformed wings, deformed body shape, and pupal cases and curled
wings attached to a deformed body were observed in adults that developed from larvae fed with apple leaves expressing Na-PI,
when compared to larvae fed with the non-transformed apple leaves. 相似文献
9.
M. E. G. Suykerbuyk P. J. Schaap H. Stam W. Musters J. Visser 《Applied microbiology and biotechnology》1995,43(5):861-870
Rhamnogalacturonase was purified from culture filtrate ofAspergillus aculeatus after growth in medium with sugar-beet pulp as carbon source. Purified protein was used to raise antibodies in mice and with
the antiserum obtained a gene coding for rhamnogalacturonase (rhgA) was isolated from a λ cDNA expression library. The clonedrhgA gene has an open-reading frame of 1320 base pairs encoding a protein of 440 amino acids with a predicted molecular mass of
45 962 Da. The protein contains a potential signal peptidase cleavage site behind Gly-18 and three potential sites forN-glycosylation. Limited homology withA. niger polygalacturonase amino acid sequences is found. A genomic clone ofrhgA was isolated from a recombinant phage λ genomic library. Comparison of the genomic and cDNA sequences revealed that the coding
region of the gene is interrupted by three introns. Furthermore, amino acid sequences of four different peptides, derived
from purifiedA. aculeatus rhamnogalacturonase, were also found in the deduced amino acid sequence ofrhgA.A. aculeatus strains overexpressing rhamnogalacturonase were obtained by cotransformation using either theA. niger pyrA gene or theA. aculeatus pyr A gene as selection marker. For expression of rhamnogalacturonase inA. awamori theA. awamori pyrA gene was used as selection marker. Degradation patterns of modified hairy regions, determined by HPLC, show the recombinant
rhamnogalacturonase to be active, and the enzyme was found to have a positive effect in the apple hot-mash liquefaction process. 相似文献
10.
Tatsuo Kakimoto 《Journal of plant research》1998,111(2):261-265
Although cytokinin plays a central role in plant development, our knowledge about the signal transduction pathway initiated
by this plant hormone is fragmentary. By randomly introducing enhancer elements into theArabidopsis genome throughAgrobacterium-mediated transformation, 5 cytokinin independent mutant calli (cki1-1, −2, −3, −4 andcki2) were obtained. These mutants exhibit typical cytokinin responses, including rapid proliferation, chloroplast differentiation,
shoot induction and inhibition of root formation, in the absence of cytokinin. TheCKl1 gene encodes a product similar to the sensor histidine kinases of two-component systems, and its overexpression in plants
induces typical cytokinin responses (Kakimoto 1996). Here I report that overexpression of this gene did not alter the auxin
reqirement ofArabidopsis. Another mutant,many shoots, which was also identified on the same screening, produced many adventitious shoots on cotyledons, petioles and true leaves.
The extended abstract of a paper presented at the 13th International Symposium in Conjugation with Award of the International
Prize for Biology “Frontier of Plant Biology” 相似文献
11.
Paolo Galli Giovanni Antonio Lodovico Broggini Markus Kellerhals Cesare Gessler Andrea Patocchi 《Molecular breeding : new strategies in plant improvement》2010,26(4):561-572
The Rvi15 (Vr2) apple scab resistance locus found in the GMAL 2473 accession has been previously mapped to the top of the Linkage Group
2 (LG2) by analyzing 89 progeny plants of a cross between ‘Idared’ and GMAL 2473. A new population of 989 progeny plants,
derived from a cross between ‘Golden Delicious’ and GMAL 2473, has been analyzed with the two SSR markers CH02c02a and CH02f06,
previously found to be associated with Rvi15 (Vr2), and with two published markers derived from NBS sequences (ARGH17 and ARGH37) estimated to map close to the Rvi15 (Vr2) locus. ARGH17 and ARGH37, were found to be the closest markers to the resistance locus, bracketing it within an interval
of 1.5 cM. The SSRs mapped one on each side of Rvi15 (Vr2). CH02f06 mapped at 2.9 cM from ARGH37 while CH02a02a mapped at 1.7 from ARGH17. The position of Rvi15 (Vr2) respect to CH02a02a indicates that Rvi15 (Vr2) and Rvi4 (Vh4), a second apple scab gene mapped on the top of LG2, are two different resistance genes. In order to develop even more tightly
linked markers to Rvi15 (Vr2), ARGH17 was used as the starting point for chromosome walking through the Rvi15 (Vr2) homolog region of the cv. ‘Florina’. A single ‘Florina’ BAC clone, 36I17, was sufficient to span the homologous locus in
the new population’s recombinant progeny. Sequencing of the 36I17 BAC clone allowed identifying seven putative ORFs, including
two showing a TIR-NBS-LRR structure. Ten additional markers could be developed mapping within a 1.8 cM interval around the
Rvi15 (Vr2) resistance gene. ARGH17 and GmTNL1 markers, the latter also derived from NBS-LRR resistance gene homolog sequence, are the
closest markers to Rvi15 (Vr2) bracketing it within a 0.5 cM interval. The availability of 12 markers within the Rvi15 (Vr2) region, all within a small physical distance (kbp) in ‘Florina’, suggests that cloning of the Rvi15 (Vr2) apple scab resistance gene from GMAL 2473 will be possible. 相似文献
12.
Krens FA Schaart JG Groenwold R Walraven AE Hesselink T Thissen JT 《Transgenic research》2011,20(5):1113-1123
Introduction of sustainable scab resistance in elite apple cultivars is of high importance for apple cultivation when aiming
at reducing the use of chemical crop protectants. Genetic modification (GM) allows the rapid introduction of resistance genes
directly into high quality apple cultivars. Resistance genes can be derived from apple itself but genetic modification also
opens up the possibility to use other, non-host resistance genes. A prerequisite for application is the long-term performance
and stability of the gene annex trait in the field. For this study, we produced and selected a series of transgenic apple
lines of two cultivars, i.e. ‘Elstar’ and ‘Gala’ in which the barley hordothionin gene (hth) was introduced. After multiplication, the GM hth-lines, non-GM susceptible and resistant controls and GM non-hth controls were planted in a random block design in a field trial in 40 replicates. Scab resistance was monitored after artificial
inoculation (first year) and after natural infection (subsequent years). After the trial period, the level of expression of
the hth gene was checked by quantitative RT-PCR. Four of the six GM hth apple lines proved to be significantly less susceptible to apple scab and this trait was found to be stable for the entire
4-year period. Hth expression at the mRNA level was also stable. 相似文献
13.
The tomato geneRSI-1 was previously identified as a molecular marker for auxin-induced lateral root initiation. We have further characterized
the expression mode of theRSI-1 gene in tomato andArabidopsis thaliana. Northern blot analyses revealed that the gene was induced specifically by auxin in tomato roots and hypocotyls. For experiments
with transgenic plants, the 5′ flanking region of theRSI-1 gene was linked to a GUS reporter gene, then transformed into tomato andArabidopsis. In these transgenic tomato plants, GUS activity was detected at the sites of initiation for lateral and adventitious roots.
Expression of the fusion gene was auxin-dependent and tissue-specific. This was consistent with results from the northern
blot analyses. In transgenicArabidopsis, the overall expression pattern of theRSI-GUS gene, including tissue specificity and auxin inducibility, was comparable to that in transgenic tomato seedlings. These results
indicate that an identical regulatory mechanism for lateral root initiation might be conserved in both plants. Thus, the expression
mode of theRSI-CUS gene inArabidopsis mutants defective in lateral root development should be investigated to provide details of this process. 相似文献
14.
A mutation in theCENTRORADIALIS (CEN) gene ofAntirrhinum and in theTERMINAL FLOWER 1 (TFL1) gene ofArabidopsis causes their indeterminate inflorescence to determinate. We clonedCEN/TFL1 homologs fromNicotiana tabacum, the wild-type of which has a determinate inflorescence. TheCEN gene was expressed in the inflorescnece meristem and kept its inflorescence meristem identity, whereas the tobacco homolog
(NCH) was expressed at a low level throughout the plant’s development. AlthoughCEN andNCH are highly homologous genes, they may have been recruited to different developmental functions during their evolution. TwoNCH genes are derived from amphidiploidN. tabacum, but both of them hybridized with its diploid parents,N. sylvestris andN. tomentosiformis. Southern blotting, and the genomic organization ofTFL1 inArabidopsis revealed that anotherCEN homolog exists in the genome ofArabidopsis. These results suggest that there are two copies of theCEN homolog per diploid plant.
The extended abstract of a paper presented at the 13th International Symposium in Conjugation with Award of the International
Prize for Biology “Frontier of Plant Biology”
These two authors contributed to this work equally. 相似文献
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Sibylle Stoeckli Karsten Mody Andrea Patocchi Markus Kellerhals Silvia Dorn 《Tree Genetics & Genomes》2009,5(1):257-267
The aim of this study was to assess the genetic basis of rust mite (Aculus schlechtendali) resistance in apple (Malus × domestica). A. schlechtendali infestation of apple trees has increased as a consequence of reduced side effects of modern fungicides on rust mites. An
analysis of quantitative trait loci (QTLs) was carried out using linkage map data available for F1 progeny plants of the cultivars ‘Fiesta’ × ‘Discovery’. Apple trees representing 160 different genotypes were surveyed for
rust mite infestation, each at three different sites in two consecutive years. The distribution of rust mites on the individual
apple genotypes was aggregated and significantly affected by apple genotype and site. We identified two QTLs for A. schlechtendali resistance on linkage group 7 of ‘Fiesta’. The AFLP marker E35M42-0146 (20.2 cM) and the RAPD marker AE10-400 (45.8 cM) were
closest positioned to the QTLs and explained between 11.0% and 16.6% of the phenotypic variability. Additionally, putative
QTLs on the ‘Discovery’ chromosomes 4, 5 and 8 were detected. The SSR marker Hi03a10 identified to be associated to one of
the QTLs (AFLP marker E35M42-0146) was traced back in the ‘Fiesta’ pedigree to the apple cultivar ‘Wagener’. This marker may
facilitate the breeding of resistant apple cultivars by marker assisted selection. Furthermore, the genetic background of
rust mite resistance in existing cultivars can be evaluated by testing them for the identified SSR marker.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
17.
Sibylle Stoeckli Karsten Mody Cesare Gessler Andrea Patocchi Mauro Jermini Silvia Dorn 《Tree Genetics & Genomes》2008,4(4):833-847
The rosy apple aphid (Dysaphis plantaginea), the leaf-curling aphid (Dysaphis cf. devecta) and the green apple aphid (Aphis pomi) are widespread pest insects that reduce growth of leaves, fruits and shoots in apple (Malus × domestica). Aphid control in apple orchards is generally achieved by insecticides, but alternative management options like growing
resistant cultivars are needed for a more sustainable integrated pest management (IPM). A linkage map available for a segregating
F1-cross of the apple cultivars ‘Fiesta’ and ‘Discovery’ was used to investigate the genetic basis of resistance to aphids.
Aphid infestation and plant growth characteristics were repeatedly assessed for the same 160 apple genotypes in three different
environments and 2 consecutive years. We identified amplified fragment length polymorphism (AFLP) markers linked to quantitative
trait loci (QTLs) for resistance to D. plantaginea (‘Fiesta’ linkage group 17, locus 57.7, marker E33M35–0269; heritability: 28.3%), and to D. cf. devecta (‘Fiesta’ linkage group 7, locus 4.5, marker E32M39–0195; heritability: 50.2%). Interactions between aphid species, differences
in climatic conditions and the spatial distribution of aphid infestation were identified as possible factors impeding the
detection of QTLs. A pedigree analysis of simple sequence repeat (SSR) marker alleles closely associated with the QTL markers
revealed the presence of the alleles in other apple cultivars with reported aphid resistance (‘Wagener’, ‘Cox’s Orange Pippin’),
highlighting the genetic basis and also the potential for gene pyramiding of aphid resistance in apple. Finally, significant
QTLs for shoot length and stem diameter were identified, while there was no relationship between aphid resistance and plant
trait QTLs.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
18.
Toshi Foster Chris Kirk William T. Jones Andrew C. Allan Richard Espley Sakuntala Karunairetnam Jasna Rakonjac 《Tree Genetics & Genomes》2007,3(3):187-197
The hormone gibberellic acid (GA) regulates growth and development throughout the plant life cycle. DELLA proteins are key
components of the GA signalling pathway and act to repress GA responses. The “DELLA” amino acid motif is highly conserved
among diverse species and is essential for GA-induced destruction of DELLA proteins, which relieves repression. Six genes
encoding the DELLA motif were identified within an apple expressed sequence tag (EST) database. Full-length cDNA clones were
obtained by RACE and these were designated MdRGL1a/b, MdRGL2a/b, and MdRGL3a/b. Sequence alignment of the predicted proteins indicates that the MdDELLAs are 37–93% homologous to one another and 44–65%
to the Arabidopsis DELLAs. The MdDELLAs cluster into three pairs, which reflect the presumed allopolyploid origins of the Maloideae. Expression analysis using quantitative real-time PCR indicates that all three pairs of MdDELLA mRNAs are expressed at the highest levels in summer arrested shoot tips and in autumn vegetative buds. Transgenic Arabidopsis expressing MdRGL2a have smaller leaves and shorter stems, take longer to flower in short days, and exhibit a reduced response to exogenous GA3, indicating significant conservation of gene function between DELLA proteins from apple and Arabidopsis.
Electronic supplementary material Supplementary material is available for this article at and is accessible for authorized users. 相似文献
19.
Common loci underlie field resistance to soybean sudden death syndrome in Forrest,Pyramid, Essex,and Douglas 总被引:8,自引:0,他引:8
Njiti VN Meksem K Iqbal MJ Johnson JE Kassem MA Zobrist KF Kilo VY Lightfoot DA 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2002,104(2-3):294-300
Soybean [Glycine max (L.) Merr.] sudden death syndrome (SDS) caused by Fusarium solani f. sp. glycines results in severe yield losses. Resistant cultivars offer the most-effective protection against yield losses but resistant
cultivars such as ’Forrest’ and ’Pyramid’ vary in the nature of their response to SDS. Loci underlying SDS resistance in ’Essex’ × Forrest
are well defined. Our objectives were to identify and characterize loci and alleles that underlie field resistance to SDS
in Pyramid×’Douglas’. SDS disease incidence and disease severity were determined in replicated field trials in six environments
over 4 years. One hundred and twelve polymorphic DNA markers were compared with SDS disease response among 90 recombinant
inbred lines from the cross Pyramid×Douglas. Two quantitative trait loci (QTLs) for resistance to SDS derived their beneficial
alleles from Pyramid, identified on linkage group G by BARC-Satt163 (261-bp allele, P=0.0005, R2=16.0%) and linkage group N by BARC-Satt080 (230-bp allele, P=0.0009, R2=15.6%). Beneficial alleles of both QTLs were previously identified in Forrest. A QTL for re- sistance to SDS on linkage group
C2 identified by BARC-Satt307 (292-bp allele, P=0.0008, R2=13.6%) derived the beneficial allele from Douglas. A beneficial allele of this QTL was previously identified in Essex. Recombinant
inbred lines that carry the beneficial alleles for all three QTLs for resistance to SDS were significantly (P≤0.05) more resistant than other recombinant inbred lines . Among these recombinant inbred lines resistance to SDS was environmentally
stable. Therefore, gene pyramiding will be an effective method for developing cultivars with stable resistance to SDS.
Received: 20 October 1999 / Accepted: 22 May 2001 相似文献
20.
A plant expression vector harboring four antifungal genes was delivered into the embryogenic calli of ‘9311’, an indica restorer
line of Super Hybrid Rice, via modified biolistic particle bombardment. Southern blot analysis indicated that in the regenerated
hygromycin-resistant plants, all the four antifungal genes, including RCH10, RAC22, β-Glu and B-RIP, were integrated into the genome of ‘9311’, co-transmitted altogether with the marker gene hpt in a Mendelian pattern. Some transgenic R1 and R2 progenies, with all transgenes displaying a normal expression level in the Northern blot analysis, showed high resistance
to Magnaporthe grisea when tested in the typical blast nurseries located in Yanxi and Sanya respectively. Furthermore, transgenic F1 plants, resulting from a cross of R2 homozygous lines with high resistance to rice blast with the non-transgenic male sterile line Peiai 64S, showed not only
high resistance to M. grisea but also enhanced resistance to rice false smut (a disease caused by Ustilaginoidea virens) and rice kernel smut (another disease caused by Tilletia barclayana). 相似文献