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1.
Molecular markers linked to the blast resistance gene <Emphasis Type="Italic">Pi-z</Emphasis> in rice for use in marker-assisted selection 总被引:6,自引:0,他引:6
Conaway-Bormans CA Marchetti MA Johnson CW McClung AM Park WD 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2003,107(6):1014-1020
Rice blast, caused by the fungal pathogen Pyricularia grisea, is a serious disease affecting rice-growing regions around the world. Current methods for identification of blast-resistant germplasm and progeny typically utilize phenotypic screening. However, phenotypic screens are influenced by environmental conditions and the presence of one resistance gene can sometimes phenotypically mask other genes conferring resistance to the same blast race. Pi-z is a dominant gene located on the short arm of chromosome 6 that confers complete resistance to five races of blast. Using sequence data found in public databases and degenerate primer pairs based on the P-loop, nucleotide binding sites and kinase domain motifs of previously cloned resistance genes, we have developed PCR-based DNA markers that cosegregate with the gene. These markers are polymorphic in a wide range of germplasm, including the narrow crosses characteristic of applied rice-breeding programs. They can now be used as a low cost, high-throughput alternative to conventional phenotypic screening for direct detection of blast resistance genes, allowing rapid introgression of genes into susceptible varieties as well as the incorporation of multiple genes into individual lines for more-durable blast resistance.Electronic Supplementary Material Supplementary material is available in the online version of this article at Communicated by D. Mackill 相似文献
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Toriba T Harada K Takamura A Nakamura H Ichikawa H Suzaki T Hirano HY 《Molecular genetics and genomics : MGG》2007,277(5):457-468
Members of the YABBY gene family have a general role that promotes abaxial cell fate in a model eudicot, Arabidopsis thaliana. To understand the function of YABBY genes in monocots, we have isolated all YABBY genes in Oryza sativa (rice), and revealed the spatial and temporal expression pattern of one of these genes, OsYABBY1. In rice, eight YABBY genes constitute a small gene family and are classified into four groups according to sequence similarity, exon-intron structure, and organ-specific expression patterns. OsYABBY1 shows unique spatial expression patterns that have not previously been reported for other YABBY genes, so far. OsYABBY1 is expressed in putative precursor cells of both the mestome sheath in the large vascular bundle and the abaxial sclerenchyma in the leaves. In the flower, OsYABBY1 is specifically expressed in the palea and lemma from their inception, and is confined to several cell layers of these organs in the later developmental stages. The OsYABBY1-expressing domains are closely associated with cells that subsequently differentiate into sclerenchymatous cells. These findings suggest that the function of OsYABBY1 is involved in regulating the differentiation of a few specific cell types and is unrelated to polar regulation of lateral organ development. 相似文献
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To clarify the diversity and function of isozymes of ascorbate peroxidase (APX) in plants, a method of producing large quantities of these proteins is needed. Here, we describe an Escherichia coli expression system for the rapid and economic expression of two rice APX genes, APXa and APXb (GeneBank accession Nos. D45423 and AB053297, respectively). The two genes were cloned into the pGEX-6p-3 vector to allow expression of APX as a glutathione-S-transferase (GST) fusion protein. The GST-APXa and GST-APXb fusion proteins were purified by affinity chromatography using a glutathione-Sepharose 4B column, with final yields of 40 and 73 mg g–1 dry cells, respectively. Specific activities were 15 and 20 mM ascorbate min–1 mg–1 protein, respectively. The Km values for ascorbate were 4 and 1 mM, respectively, and those for H2O2 were 0.3 and 0.7 mM, respectively indicating that the two rice isoenzymes have different properties.Revisions requested 27 September 2004; Revisions received 12 November 2004 相似文献
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The SNAP25-type proteins belong to the superfamily of the SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors), and function as important components of the vesical trafficking machinery in eukaryotic cells. In this paper, we report the cloning and expression characterization of OsSNAP32 gene, and the subcellular localization of its encoded protein. The OsSNAP32 gene contains five exons and four introns, and is located between RFLP markers C12276S and S1917 on chromosome 2 in rice. The OsSNAP32 has a molecular weight of 31.3 kD, comprises 283 amino acid residues, and contains Qb-SNARE and Qc-SNARE domains in the N- and C-terminal, respectively. Multiple sequence alignment of the SNARE domains indicates that OsSNAP32 protein is homologous to HvSNAP34 and HvSNAP28 (63% and 55% of amino acid identity respectively) from barley. The transient expression method in onion epidermal cells, revealed that OsSNAP32 is located in the plasma membrane, like other SNAP25-type proteins. Semi-quantitative RT-PCR assay showed that the OsSNAP32 is highly expressed in leaves and culms, and low in roots of rice, while hardly detected in immature spikes and flowering spikes. The expression of OsSNAP32 was significantly activated in rice seedlings treated with H2O2, PEG6000, and low temperature or after inoculation with rice blast (Magnaporthe grisea strain Hoku 1). The results suggest that this gene belongs to a novel member of this gene family encoding SNAP25-type proteins, involved in the rice responses to biotic and abiotic stresses. 相似文献
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Genetic and physical mapping of <Emphasis Type="Italic">Pi36</Emphasis>(t), a novel rice blast resistance gene located on rice chromosome 8 总被引:12,自引:0,他引:12
Blast resistance in the indica cultivar (cv.) Q61 was inherited as a single dominant gene in two F2 populations, F2-1 and F2-2, derived from crosses between the donor cv. and two susceptible japonica cvs. Aichi Asahi and Lijiangxintuanheigu (LTH), respectively. To rapidly determine the chromosomal location of the resistance
(R) gene detected in Q61, random amplified polymorphic DNA (RAPD) analysis was performed in the F2-1 population using bulked-segregant analysis (BSA) in combination with recessive-class analysis (RCA). One of the three linked
markers identified, BA1126550, was cloned and sequenced. The R gene locus was roughly mapped on rice chromosome 8 by comparison of the BA1126550 sequence with rice sequences in the databases (chromosome landing). To confirm this finding, seven known markers, including
four sequence-tagged-site (STS) markers and three simple-sequence repeat (SSR) markers flanking BA1126550 on chromosome 8, were subjected to linkage analysis in the two F2 populations. The locus was mapped to a 5.8 cM interval bounded by RM5647 and RM8018 on the short arm of chromosome 8. This
novel R gene is therefore tentatively designated as Pi36(t). For fine mapping of the Pi36(t) locus, five additional markers including one STS marker and four candidate resistance gene (CRG) markers were developed
in the target region, based on the genomic sequence of the corresponding region of the reference japonica cv. Nipponbare. The Pi36(t) locus was finally localized to an interval of about 0.6 cM flanked by the markers RM5647 and CRG2, and co-segregated with
the markers CRG3 and CRG4. To physically map this locus, the Pi36(t)-linked markers were mapped by electronic hybridization to bacterial artificial chromosome (BAC) or P1 artificial chromosome
(PAC) clones of Nipponbare, and a contig map was constructed in silico through Pairwise BLAST analysis. The Pi36(t) locus was physically delimited to an interval of about 17.0 kb, based on the genomic sequence of Nipponbare. 相似文献
10.
NADP-malic enzyme (NADP-ME, EC 1.1.1.40) functions in many different pathways in plant and may be involved in plant defense
such as wound and UV-B radiation. Here, expression of the gene encoding cytosolic NADP-ME (cytoNADP-ME, GenBank Accession No. AY444338) in rice (Oryza sativa L.) seedlings was induced by salt stress (NaCl). NADP-ME activities in leaves and roots of rice also increased in response
to NaCl. Transgenic Arabidopsis plants over-expressing rice cytoNADP-ME had a greater salt tolerance at the seedling stage than wild-type plants in MS medium-supplemented with different levels
of NaCl. Cytosolic NADPH/NADP+ concentration ratio of transgenic plants was higher than those of wild-type plants. These results suggest that rice cytoNADP-ME confers salt tolerance in transgenic Arabidopsis seedlings. 相似文献
11.
Liu X Yang Q Lin F Hua L Wang C Wang L Pan Q 《Molecular genetics and genomics : MGG》2007,278(4):403-410
Blast, caused by the ascomycete fungus Magnaporthe oryzae, is one of the most devastating diseases of rice worldwide. The Chinese native cultivar (cv.) Q15 expresses the broad-spectrum
resistance to most of the isolates collected from China. To effectively utilize the resistance, three rounds of linkage analysis
were performed in an F2 population derived from a cross of Q15 and a susceptible cv. Tsuyuake, which segregated into 3:1 (resistant/susceptible)
ratio. The first round of linkage analysis employing simple sequence repeat (SSR) markers was carried out in the F2 population through bulked-segregant assay. A total of 180 SSR markers selected from each chromosome equally were surveyed.
The results revealed that only two polymorphic markers, RM247 and RM463, located on chromosome 12, were linked to the resistance
(R) gene. To further define the chromosomal location of the R gene locus, the second round of linkage analysis was performed using additional five SSR markers, which located in the region
anchored by markers RM247 and RM463. The locus was further mapped to a 0.27 cM region bounded by markers RM27933 and RM27940
in the pericentromeric region towards the short arm. For fine mapping of the R locus, seven new markers were developed in the smaller region for the third round of linkage analysis, based on the reference
sequences. The R locus was further mapped to a 0.18 cM region flanked by marker clusters 39M11 and 39M22, which is closest to, but away from
the Pita/Pita
2 locus by 0.09 cM. To physically map the locus, all the linked markers were landed on the respective bacterial artificial
chromosome clones of the reference cv. Nipponbare. Sequence information of these clones was used to construct a physical map
of the locus, in silico, by bioinformatics analysis. The locus was physically defined to an interval of ≈37 kb. To further
characterize the R gene, five R genes mapped near the locus, as well as 10 main R genes those might be exploited in the resistance breeding programs, were selected for differential tests with 475 Chinese
isolates. The R gene carrier Q15 conveys resistances distinct from those conditioned by the carriers of the 15 R genes. Together, this valuable R gene was, therefore, designated as Pi39(t). The sequence information of the R gene locus could be used for further marker-based selection and cloning.
Xinqiong Liu and Qinzhong Yang contributed equally to this work. 相似文献
12.
de Wilde C Uzan E Zhou Z Kruus K Andberg M Buchert J Record E Asther M Lomascolo A 《Transgenic research》2008,17(4):515-527
Laccases have numerous biotechnological applications, among them food processing. The widespread use of laccases has increased the demand for an inexpensive and safe source of recombinant enzyme. We explored the use of a rice-based system for the production of two fungal laccases derived from the ascomycete Melanocarpus albomyces and the basidiomycete Pycnoporus cinnabarinus. High-expression levels of active recombinant laccases were achieved by targeting expression to the endosperm of rice seeds. The laccase cDNAs were fused to a plant-derived signal sequence for targeting to the secretory pathway, and placed under the control of a constitutive seed-specific promoter fused to an intron for enhanced expression. This construct enabled the recovery of on average 0.1-1% of soluble laccase in total soluble proteins (TSP). The highest yields of recombinant laccases obtained in rice seeds were 13 and 39 ppm for riceMaL and ricePycL, respectively. The rice-produced laccases were purified and characterized. The wild-type and the recombinant proteins showed similar biochemical features in terms of molecular mass, pI, temperature and optimal pH and the N-terminus was correctly processed. Although presenting lower kinetic parameters, the rice-produced laccases were also suitable for the oxidative cross-linking of a food model substrate [maize-bran feruloylated arabinoxylans (AX)]. 相似文献
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Kobayashi S Fukuta Y Sato T Osaki M Khush GS 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2003,107(8):1350-1356
Rice (Oryza sativa L.) plants develop vertically with shoot elongation and horizontally with tillering. The purpose of this study was to identify and characterize genomic regions influencing the rice plant architecture by quantitative trait locus (QTL) analysis for the component traits: culm length (CL), panicle length (PnL), panicle number (PnN) and tiller number (TN). For this QTL analysis, 191 recombinant inbred lines (F7) derived from a cross of Milyang 23 (M23) and Akihikari (AK) were grown in 1995, 1996 and 1997 (May–Oct) in Joetsu, Japan (temperate climate), and in the 2000 dry season (Jan–Apr), the 2000 wet season (Jun–Oct) and the 2001 dry season in Los Baños, The Philippines (tropical climate). Results showed that rice plant architecture was influenced by 19 genomic regions categorized into five groups. In Group I, two regions (on chrs. 6 and 11) affected shoot elongation (CL and PnL) and tillering (PnN and TN) in opposite directions more significantly in Los Baños than in Joetsu. In Group II, two regions (chrs. 3 and 12) affected shoot elongation, whereas in Group III, five regions [chrs. 1 (two), 2, 3 and 9] affected only culm length (CL). Expressions of four regions of Group III were influenced by either tropical or temperate environments. In Group IV, seven regions (chrs. 1, 2, 4, 5, 6, 8 and 9) controlled panicle development (PnN or PnL), and in Group V, three regions (chrs. 1, 2 and 3) regulated tillering (PnN or TN). Characterizing these 19 genomic regions provided a detailed analysis of rice plant architecture with emphasis on the multiple effect and environmental responsive regions.Communicated by D. Mackill 相似文献
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Berruyer R Adreit H Milazzo J Gaillard S Berger A Dioh W Lebrun MH Tharreau D 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2003,107(6):1139-1147
Rice blast disease is a major constraint for rice breeding. Nevertheless, the genetic basis of resistance remains poorly understood for most rice varieties, and new resistance genes remain to be identified. We identified the resistance gene corresponding to the cloned avirulence gene ACE1 using pairs of isogenic strains of Magnaporthe grisea differing only by their ACE1 allele. This resistance gene was mapped on the short arm of rice chromosome 8 using progenies from the crosses IR64 (resistant) × Azucena (susceptible) and Azucena × Bala (resistant). The isogenic strains also permitted the detection of this resistance gene in several rice varieties, including the differential isogenic line C101LAC. Allelism tests permitted us to distinguish this gene from two other resistance genes [Pi11 and Pi-29(t)] that are present on the short arm of chromosome 8. Segregation analysis in F2 populations was in agreement with the existence of a single dominant gene, designated as Pi33. Finally, Pi33 was finely mapped between two molecular markers of the rice genetic map that are separated by a distance of 1.6 cM. Detection of Pi33 in different semi-dwarf indica varieties indicated that this gene could originate from either one or a few varieties.Communicated by D.J. Mackill 相似文献
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Two rice cytosolic ascorbate peroxidases differentially improve salt tolerance in transgenic <Emphasis Type="Italic">Arabidopsis</Emphasis> 总被引:1,自引:0,他引:1
In order to determine the different roles of rice (Oryza sativa L.) cytosolic ascorbate peroxidases (OsAPXa and OsAPXb, GenBank accession nos. D45423 and AB053297, respectively) under salt stress, transgenic Arabidopsis plants over-expressing OsAPXa or OsAPXb were generated, and they all exhibited increased tolerance to salt stress compared to wild-type plants. Moreover, transgenic
lines over-expressing OsAPXb showed higher salt tolerance than OsAPXa transgenic lines as indicated by root length and total chlorophyll content. In addition to ascorbate peroxidase (APX) activity,
antioxidant enzyme activities of catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR), which are also
involved in the salt tolerance process, and the content of H2O2 were also assayed in both transgenic and wild-type plants. The results showed that the overproduction of OsAPXb enhanced and maintained APX activity to a much higher degree than OsAPXa in transgenic Arabidopsis during treatment with different concentrations of NaCl, enhanced the active oxygen scavenging system, and protected plants
from salt stress by equilibrating H2O2 metabolism. Our findings suggest that the rice cytosolic OsAPXb gene has a more functional role than OsAPXa in the improvement of salt tolerance in transgenic plants.
Zhenqiang Lu and Dali Liu contributed equally. 相似文献
16.
Zhen Wang Changbin Chen Yunyuan Xu Rongxi Jiang Ye Han Zhihong Xu Kang Chong 《Plant Molecular Biology Reporter》2004,22(4):409-417
In the last decade, RNA interferences (RNAi) has proven to be an effective strategy to knock out homologous genes in a wide
range of species. Based on its principle, a new generation of vectors containing an inverted target sequence separated by
an intron as a loop, developing simplifications to the procedure of RNAi construction are required to improve the efficiency
of gene inactivation techniques. Here, a novel polymerase chain reaction (PCR)—based RNAi vector pTCK303 with a maize ubiquitin
promoter, 2 specific multiple enzyme sites, and a rice intron was constructed for monocot gene silencing. With this vector,
only 1 PCR product amplified by a single pair of primers and 2 ligation reactions were needed to create an RNAi construct,
which shortened the time span before being transformed into the plant. To test the efficiency of vector pTCK303, a rice geneOsGAS1 was used, and its RNAi construct was introduced into rice calli. Southern blot analysis of the transgenic rice confirmed
the presence of theOsGAS1 RNAi structure. The decrease inOsGAS1 level in the transgenic rice was detected by Northern blot probed with anOsGAS1-specific sequence. Moreover, the rate of inhibition of the RNA expression level in RNAi transgenic rice was approximately
85% according to our real-time PCR. Therefore, the RNAi vector pTCK303 based on the homology-dependent gene-silencing mechanisms
facilitated the inhibition of endogenous genes in a monocot and was proven to be a practical and efficient platform for silencing
a rice gene.
These authors contributed equally to this work. 相似文献
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Shuifeng Ye Lei Wang Weibo Xie Bingliang Wan Xianghua Li Yongjun Lin 《Plant molecular biology》2009,70(3):311-325
Calcium-dependent protein kinases (CDPKs) control plant development and response to various stress environments through the
important roles in the regulation of Ca2+ signaling. Thirty-one CDPK genes have been identified in the rice genome by a complete search of the genome based upon HMM profiles. In this study,
the expression of this gene family was analyzed using the Affymetrix rice genome array in three rice cultivars: Minghui 63,
Zhenshan 97, and their hybrid Shanyou 63 independently. Twenty-seven tissues sampled throughout the entire rice life-span
were studied, along with three hormone treatments (GA3, NAA and KT), applied to the seedling at the trefoil stage. All 31
genes were found to be expressed in at least one of the experimental stages studied and revealed diverse expression patterns.
We identified differential expression of the OsCPK genes in the stamen (1 day before flowering), the panicle (at the heading stage), the endosperm (days after pollination)
and also in callus, in all three cultivars. Eight genes, OsCPK2, OsCPK11, OsCPK14, OsCPK22, OsCPK25, OsCPK26, OsCPK27 and OsCPK29 were found dominantly expressed in the panicle and the stamen, and five genes, OsCPK6, OsCPK7, OsCPK12, OsCPK23 and OsCPK31 were up-regulated in the endosperm stage. The OsCPK genes were also found to be regulated in rice seedlings subjected to different hormone treatment conditions, however their
expression were not the same for all varieties. These diverse expression profiles trigger the functional analysis of the CDPK family in rice.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
18.
Li S Li T Kim WD Kitaoka M Yoshida S Nakajima M Kobayashi H 《Biotechnology letters》2007,29(4):635-640
The putative raffinose synthase gene from rice was cloned and expressed in Escherichia coli. The enzyme displayed an optimum activity at 45°C and pH 7.0, and a sulfhydryl group was required for its activity. The enzyme
was specific for galactinol and p-nitrophenyl-α-d-galactoside as galactosyl donors, and sucrose, lactose, 4−β-galactobiose, N-acetyl-d-lactosamine, trehalose and lacto-N-biose were recognized as galactosyl acceptors. 相似文献
19.
Guosheng Shao Mingxue Chen Weixia Wang Guoping Zhang 《Journal of Plant Growth Regulation》2008,27(3):205-210
The influence of betaine aldehyde dehydrogenase (BADH) and salinity pretreatment on oxidative stress under cadmium (Cd) toxicity
was investigated in rice cv. Xiushui 11 and its BADH-transgenic line Bxiushui 11. The results showed that plants previously treated with 4.25 and 8.5 mM NaCl, respectively, for
5 days each had higher Cd concentrations in both roots and shoots of the two rice genotypes compared with the controls. Malondialdehyde
(MDA) content in both leaves and roots was increased by salinity pretreatment and was significantly lower in the salinity-pretreatment
plants than in the controls when the plants were consequently exposed to Cd stress. Salinity pretreatment also increased proline
content and the activities of superoxide dismutase (SOD) and peroxidase (POD) in both leaves and roots. It can be assumed
that salinity pretreatment enhances the defensive ability of plants against oxidative stress through increasing activities
of antioxidative enzymes. The BADH-transgenic line (Bxiushui 11) had lower Cd and MDA content, higher SOD and POD activities, and higher proline content than
its wild type (Xiushui 11). The current results suggest that betaine, a product of BADH expression, improves the tolerance of rice plants to Cd stress through increasing the activities of antioxidative enzymes
and osmoprotectant content. 相似文献
20.
Ventelon-Debout M Nguyen TT Wissocq A Berger C Laudie M Piégu B Cooke R Ghesquière A Delseny M Brugidou C 《Molecular genetics and genomics : MGG》2003,270(3):253-262
Several cDNA libraries were constructed using mRNA isolated from roots, panicles, cell suspensions and leaves of non-stressed Oryza sativa indica (IR64) and japonica (Azucena) plants, from wounded leaves, and from leaves of both cultivars inoculated with Rice Yellow Mottle Virus (RYMV). A total of 5549 cleaned expressed sequence tags (ESTs) were generated from these libraries. They were classified into functional categories on the basis of homology, and analyzed for redundancy within each library. The expression profiles represented by each library revealed great differences between indica and japonica backgrounds. EST frequencies during the early stages of RYMV infection indicated that changes in the expression of genes involved in energy metabolism and photosynthesis are differentially accentuated in susceptible and partially resistant cultivars. Mapping of these ESTs revealed that several co-localize with previously described resistance gene analogs and QTLs (quantitative trait loci). 相似文献