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Title: Gene expression in grass ovaries infected with seed born fungal endophyte Neotyphodium occultans analyzed by a next-generation sequencing system

Author
item SUGAWARA, KOYA - National Agricultural Research Organization - Japan (NARO)
item NAGABHYYRU, PADMAJA - University Of Kentucky
item Dinkins, Randy
item SCHARDL, CHRISTOPHER - University Of Kentucky
item JAROMCZYK, JOLANTA - University Of Kentucky
item ARAKAWA, AKIRA - National Agricultural Research Organization - Japan (NARO)
item SHIBA, TAKUYA - National Agricultural Research Organization - Japan (NARO)
item OKABE, IKUKO - National Agricultural Research Organization - Japan (NARO)
item TSUKIBOSHI, TAKAO - National Agricultural Research Organization - Japan (NARO)

Submitted to: Mycological Society Of Japan
Publication Type: Abstract Only
Publication Acceptance Date: 7/18/2011
Publication Date: 2/23/2012
Citation: Sugawara, K., Nagabhyyru, P., Dinkins, R.D., Schardl, C.L., Jaromczyk, J.W., Arakawa, A., Shiba, T., Okabe, I., Tsukiboshi, T. 2012. Gene expression in grass ovaries infected with seed born fungal endophyte Neotyphodium occultans analyzed by a next-generation sequencing system. Mycological Society Of Japan. C15.

Interpretive Summary:

Technical Abstract: Fungal endophytes of the genus Neotyphodium form symbiotic associations with many grass species of the subfamily Pooideae, including some important forage and turf species such as Lolium grasses. The endophytes are maintained in host plant communities by seed transmission from maternal plants to offsprings, but physiology of their invasion and settling in host seeds are yet unknown. To elucidate this important process, gene expression in infected grass ovaries were analyzed using Italian ryegrass (Lolium multiflorum) infected with N. occultans, by a next-generation sequencing system. A seed set from an infected plant were divided into two groups, and one of it was heat treated mildly (43°C 15min + 57°C x 40min) to kill the endophyte. After that, these seeds were grown to flower in a glasshouse with occasional checking by microscopy for their infection status. Immature ovaries were collected from selected five infected and two endophyte free plants, and pooled by their infection status and mRNA were extracted. From these two mRNA samples, the sequencing system obtained 27221673 reads from endophyte infected, and 25910556 reads from endophyte free ones. The sequence reads were assembled into 86468 contigs (minimum contig length set to 200 bp). Several hundred contigs were found to be differentially expressed between the endophyte infected and endophyte free plants. Experiments are ongoing to verify differential expression and identify the gene products in relation with genome information reported from related plant and fungi species.