|Li, W. - AGRON DEPT, NANJING UNIV|
|Muthukrishnan, S. - BIOCHEM, KSU, MANHATTAN,|
|Liu, D. - AGRON DEPT, NANJING UNIV|
|Chen, P. - AGRON DEPT, NANJING UNIV|
|Gill, Bikram - PLANT PATH, KSU, MANHATTA|
Submitted to: Journal of Theoretical and Applied Genetics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: March 1, 2001
Publication Date: May 20, 2001
Citation: Li, W.L., Faris, J.D., Muthukrishnan, S., Liu, D.J., Chen, P.D., Gill, B.S. 2001. Isolation and characterization of novel cdna clones of acidic chitinases and beta-1,3-glucanases from wheat spikes infected by fusarium graminearum. Journal of Theoretical and Applied Genetics 102:353-362. Interpretive Summary: In response to a pathogen attack, plants activate genes that produce enzymes involved in combatting the pathogen. Two such defense response enzymes are chitinase and beta-1,3-glucanase. Fusarium head blight, or wheat scab, is caused by the fungus Fusarium graminearum and is an important disease of wheat. Very few wheat varieties possess resistance to Fusarium head blight, but the Chinese variety Sumai-3 has relatively high levels of resistance compared to most. To determine if defense response genes are involved in conditioning resistance to Fusarium head blight in Sumai-3, we cloned and characterized genes that encode chitinase and beta-1,3-glucanase from Sumai-3. Two genes representing two different classes of chitinases and two genes representing two different forms of beta-1,3-glucanase were isolated, and their DNA sequences and chromosomal locations were determined. Further analysis indicated that the expression of these genes was induced in wheat spikes upon infection with Fusarium graminearum indicating that they are involved in the defense response. Furthermore, the chitinase and beta-1,3-glucanase enzymes were produced more rapidly in Sumai-3 compared to a susceptible variety within the first 24 hours following infection by the fungus. This is the first report of the induction of these two classes of chitinases in cereals by a fungal pathogen.
Technical Abstract: Chitinases and beta-1,3-glucanases are important components of plant defense in response to attack by pathogens. To identify specific chitinases and beta-1,3-glucanases, we constructed a cDNA library using mRNA from wheat spikelets inoculated with conidia of Fusarium graminearum. Two chitinase and two beta-1,3-glucanase clones were isolated using a rice chitinase Ia gene and barley cDNA clones for chitinase II and beta-1,3-glucanase as probes. Sequence analysis showed that the cDNA clone SM194 encodes an acidic isoform of class VII chitinase, the cDNA clone SM383 encodes a class IV chitinase and the cDNA clones SM289 and SM638 encode two different acidic isoforms of beta-1,3-glucanase. Nulli-tetrasomic analysis indicated that SM194 and SM383 were located on all of the group-2 chromosomes of wheat. Genetic mapping showed that at least three copies of class-IV and/or class-VII chitinase genes were clustered on the long arm of chromosome 2D of Aegilops tauschii and that they mapped genetically close to the centromere. SM289 and SM638 were located on all of the group-3 chromosomes of wheat by nulli-tetrasomic analysis, and to the beta-1,3-glucanase clusters in the 3BL and 3DL chromosome arms of wheat by genetic mapping. Northern blot hybridization showed that the expression of these genes is induced upon infection with Fusarium graminearum. The accumulation of transcripts for these PR-proteins was more rapid in the resistant variety Sumai 3 than in its susceptible mutant during the first 24 h. This is the first report of the induction of class-IV and class-VII chitinases in cereals by a fungal pathogen.