Identification of a protein secreted by the blister rust fungus Cronartium ribicola in infected white pines and its cDNA cloning and characterization

1999 ◽  
Vol 77 (6) ◽  
pp. 800-808
Author(s):  
Abul Kalam Mohammed Ekramoddoullah ◽  
Yingchun Tan ◽  
Xueshu Yu ◽  
Doug William Taylor ◽  
Santosh Misra
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Protein Sequence ◽  
Rust Fungus ◽  
Major Protein ◽  
Protein Sequence Analysis ◽  
White Pine ◽  
Blister Rust ◽  
Pine Seedlings ◽  
Sugar Pine

Previously we showed that a white pine protein Pin m III (a member of PR10 family of pathogenesis-related proteins) is up-regulated by infection in the white pine blister rust pathosystem. In this study, a blister rust fungal protein, Cro r I, which is similar in size to Pin m III (19 kDa), was detected in the infected white pine tissues. The N-terminal amino acid sequence of Cro r I isolated from infected pine foliage and from fungal mycelia was identical. Rabbit antibody was prepared to a synthetic N-terminal peptide and was purified by immunoaffinity. The purified antibody was used in a Western immunoblot to quantify the amount of Cro r I in various tissues. In western white pine seedlings the amount of Cro r I was significantly (p < 0.0001) higher in infected tissues of cankered seedlings than the infected tissues of resistant seedlings. In sugar pine seedlings, the amount of Cro r I was also significantly (p < 0.01) higher in infected tissues of susceptible seedlings than in resistant seedlings. Furthermore, Cro r I is secreted by the blister rust fungus and was found to be translocated to the healthy tissues of cankered white pines. Cro r I is a major protein that could be extracted from infected foliage by vacuum infiltration. The level of Cro r I detected in the mycelium of different isolates varied. The cDNA of Cro r I was isolated by reverse transcription - polymerase chain reaction. Comparison of the DNA sequence and the deduced protein sequence with data bases revealed that it is a previously undescribed protein. The calculated molecular weight from the deduced protein sequence of Cro r I was 16.7 kDa and the calculated isoelectric point was 9.55. Protein sequence analysis showed that Cro r I has two potential N-linked glycosylation sites in its sequence.Key words: translocation, elicitor, antibody, amino acid sequence.

scholarly journals Amino acid sequence of Fel dI, the major allergen of the domestic cat: protein sequence analysis and cDNA cloning.

1991 ◽  
Vol 88 (21) ◽  
pp. 9690-9694 ◽  
Author(s):  
J. P. Morgenstern ◽  
I. J. Griffith ◽  
A. W. Brauer ◽  
B. L. Rogers ◽  
J. F. Bond ◽  
...  
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Amino Acid Sequence ◽  
Cdna Cloning ◽  
Protein Sequence ◽  
Major Allergen ◽  
Domestic Cat ◽  
Protein Sequence Analysis

Characterisation of a fall protein of sugar pine and detection of its homologue associated with frost hardiness of western white pine needles

1995 ◽  
Vol 25 (7) ◽  
pp. 1137-1147 ◽  
Author(s):  
Abul K.M. Ekramoddoullah ◽  
Doug Taylor ◽  
Barbara J. Hawkins
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Pine Needles ◽  
Frost Hardiness ◽  
White Pine ◽  
Terminal Amino Acid ◽  
Sugar Pine ◽  
Western White Pine ◽  
Terminal Amino ◽  
Terminal Amino Acid Sequence

A 19-kDa protein of sugar pine (named Pinl I; i.e., protein I of Pinuslambertiana Dougl.) was detected in increasing amounts in the fall. This protein was separated by SDS–PAGE and also by two-dimensional gel electrophoresis. Pinl I was composed of two acidic isoforms. This protein was subjected to N-terminal amino acid sequence analysis. The two isoforms had an identical N-terminal amino acid sequence. The N-terminal peptide was synthesized and purified, and the purity of the synthetic peptide was greater than 95%. The peptide was conjugated to a carrier protein, keyhole limpet hemocyanin (KLH). Rabbits were immunized with peptide–KLH and the antipeptide antibody was purified from the crude antisera by immunoaffinity chromatography. The antibody was shown to bind specifically to PinI I. This anti-Pin I I antibody was used in a Western immunoblot to detect the homologues of Pin1 1 in two other five-needled pines: western white pine (Pinusmonticola Dougl.; named Pinm III) and eastern white pine (Pinusstrobus L.). The antibody was also used to monitor the seasonal variation of Pinm III in western white pine needles. Pinm III was shown to be associated with overwintering of western white pine seedlings. A significant correlation was observed between the frost hardiness of western white pine foliage and the content of Pinm III.

Cotyledon test for major gene resistance to white pine blister rust in sugar pine

1980 ◽  
Vol 58 (17) ◽  
pp. 1912-1914 ◽  
Author(s):  
Bohun B. Kinloch Jr. ◽  
Mardi Comstock
Keyword(s):  
Major Gene ◽  
White Pine Blister Rust ◽  
White Pine ◽  
Blister Rust ◽  
Pine Seedlings ◽  
Major Gene Resistance ◽  
Sugar Pine ◽  
Time Required ◽  
Resistant Genotypes

Major gene resistance (hypersensitivity) to white pine blister rust can be detected on cotyledons of inoculated sugar pine seedlings shortly after germination. The cotyledon test reduces the time required for evaluating resistant genotypes from a few years to a few weeks.

THE RELATION OF GROWTH REGULATORS TO THE DEVELOPMENT OF SYMPTOMS AND THE EXPRESSION OF STEM RESISTANCE IN WHITE PINE INFECTED WITH BLISTER RUST

1967 ◽  
Vol 45 (4) ◽  
pp. 501-513 ◽  
Author(s):  
Michael G. Boyer
Keyword(s):  
Gibberellic Acid ◽  
Growth Regulators ◽  
Indoleacetic Acid ◽  
Rust Fungus ◽  
Gall Formation ◽  
White Pine ◽  
Anatomical Studies ◽  
Blister Rust ◽  
Pine Seedlings ◽  
Stem Resistance

The application of the growth regulators indoleacetic acid and gibberellic acid to stems of white pine seedlings produced many symptoms characteristic of the disease caused by the blister rust fungus, thus suggesting their possible involvement in gall formation. When a kinin-like compound, benzimidazole, was applied, a wound periderm was initiated in seedlings infected with the blister rust fungus but not in healthy seedlings. This effect was augmented by indoleacetic acid and gibberellic acid. It seems to be a phenomenon very similar to that observed in stem-resistant white pine, and a possible relationship is proposed based on anatomical studies.

scholarly journals Chitin-binding proteins in potato (Solanum tuberosum L.) tuber. Characterization, immunolocalization and effects of wounding

1992 ◽  
Vol 283 (3) ◽  
pp. 813-821 ◽  
Author(s):  
D J Millar ◽  
A K Allen ◽  
C G Smith ◽  
C Sidebottom ◽  
A R Slabas ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Binding Proteins ◽  
Solanum Tuberosum L ◽  
Major Protein ◽  
Post Translational Modification ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Terminal Amino Acid Sequence

Tubers of potato (Solanum tuberosum L.) contain a number of chitin-binding proteins which have possible functions in defence against pathogens. A major protein of the tuber is the chitin-binding lectin which has been further characterized with respect to its antigenicity and N-terminal amino acid sequence. By using an antiserum monospecific for tuber lectin in unwounded potato the protein was found in the cytoplasm and vacuole, unusually for a hydroxyproline-rich glycoprotein, but consistent with its soluble nature in subcellular extracts. Little increased synthesis of the lectin precursor or the post-translationally modified form could be demonstrated in excised potato tuber discs. However, after wounding there is increased synthesis of another hydroxyproline-containing glycoprotein of Mr 57,000, which binds to chitin and shares common epitopes with the lectin. In comparison with the tuber lectin, this novel glycoprotein contains less hydroxyproline, but from its overall composition it is clearly not an underhydroxylated form of the tuber lectin. It differed in its N-terminal amino acid sequence and was much less glycosylated, although arabinose was still present. Synthesis of the Mr-57,000 polypeptide began after the initial burst of protein synthesis and increased, reaching a peak at 24 h after wounding. The protein was produced with its enzymes of post-translational modification, prolyl hydroxylase and arabinosyltransferase, concomitantly with the marker enzymes for wounding, phenylalanine ammonia-lyase and membrane-bound phenol oxidase and peroxidase.

scholarly journals Use of triadimefon to control white pine blister rust

1996 ◽  
Vol 72 (6) ◽  
pp. 637-638 ◽  
Author(s):  
Jean A. Bérubé
Keyword(s):  
Cronartium Ribicola ◽  
White Pine Blister Rust ◽  
White Pine ◽  
Growth Season ◽  
Effective Protection ◽  
Rust Infection ◽  
Blister Rust ◽  
Pine Seedlings ◽  
Second Growth ◽  
Bare Root

White pine seedlings were treated with triadimefon two weeks prior to natural inoculation with Cronartium ribicola and were observed for two growth seasons. During the second growth season in the greenhouse the incidence of blister rust symptoms was 70.8% for the untreated controls, whereas only 3.8% of the treated seedlings showed symptoms of blister rust. Triadimefon offers effective protection against white pine blister rust infection and would enable the production of bare root seedlings in areas prone to blister rust infection.

scholarly journals Genetic Specificity in the White Pine-Blister Rust Pathosystem

2002 ◽  
Vol 92 (3) ◽  
pp. 278-280 ◽  
Author(s):  
Bohun B. Kinloch ◽  
Gayle E. Dupper
Keyword(s):  
North America ◽  
Great Basin ◽  
Single Gene ◽  
Low Frequency ◽  
White Pine Blister Rust ◽  
White Pine ◽  
Mendelian Segregation ◽  
Blister Rust ◽  
Sugar Pine ◽  
Bristlecone Pine

Four of eight white pine species native to western North America surveyed for resistance to white pine blister rust by artificial inoculation showed classical hypersensitive reactions (HR) at frequencies ranging from very low to moderate. Mendelian segregation, indicating a single dominant allele for resistance (Cr3), was observed in southwestern white pine (Pinus strobiformis), as it was previously in sugar pine (P. lambertiana, Cr1) and western white pine (P. monticola, Cr2). HR was present at a relatively high frequency (19%) in one of five bulk seed lot sources of limber pine (P. flexilis), and was also presumed to be conditioned by a single gene locus, by analogy with the other three species. HR was not found in whitebark pine (P. albcaulis), Mexican white pine (P. ayacahuite), foxtail pine (P. balfouriana), or Great Basin bristlecone pine (P. longaeva), but population and sample sizes in these species may have been below the level of detection of alleles in low frequency. When challenged by (haploid) inocula from specific locations known to harbor virulence to Cr1 or Cr2, genotypes carrying these alleles and Cr3 reacted differentially, such that inoculum virulent to Cr1 was avirulent to Cr2, and inoculum virulent to Cr2 was avirulent to Cr1. Neither of these two inocula was capable of neutralizing Cr3. Although blister rust traditionally is considered an exotic disease in North America, these results, typical of classic gene-for-gene interactions, suggest that genetic memory of similar encounters in past epochs has been retained in this pathosystem.

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