Virulence of Leptographium serpens on Longleaf Pine Seedlings Under Varying Soil Moisture Regimes.

Recently, Leptographium serpens has been recovered from the roots of declining and dead longleaf pine (Pinus palustris) in stands associated with various abiotic stresses. Although most data suggest that L. serpens is pathogenic to various Pinus spp., there is little known of its virulence on longleaf pine or its relationship with abiotic stress in causing disease. These trials examined the effects of L. serpens infection coupled with drought stress. Trials began with wound inoculations of bareroot longleaf pine seedlings in spring 2006 and 2007 at the seedling stress facility at Auburn University. Soon after inoculation, seedlings were also subjected to adequate moisture, moderate drought, or severe drought. Sixteen weeks after inoculation, longleaf pine survival, L. serpens virulence, and seedling growth characteristics were measured. Longleaf pine seedlings inoculated with L. serpens had 33% mortality (138/420) which was significantly greater than nonwounded control seedlings (22%, 47/211). Survival and lesion size on longleaf pine suggests that L. serpens is moderately pathogenic to longleaf pine seedlings. Separately, moisture stress associated with low soil moisture also contributed to seedling mortality. Results suggest that L. serpens infection and moisture stress commonly experienced by southern pines act independently to stress longleaf pine.

Pathogenicity of Leptographium Species Associated with Loblolly Pine Decline.

Freshly lifted seedlings and 21-year-old trees of loblolly pine were wound-inoculated with Leptographium species recovered from the soil and/or roots of trees with loblolly decline symptoms in central Alabama. Seedlings inoculated with L. procerum in the greenhouse produced significantly fewer root initials and a smaller root mass than control seedlings. Vertical lesions produced in seedlings by L. serpens and L. terebrantis were significantly longer than in controls. Lesions produced in mature trees by L. serpens and L. lundbergii were significantly longer than in controls. Of the fungi tested, L. serpens, L. terebrantis, and L. lundbergii were the most aggressive and may pose the greatest threat to loblolly pines.

Tobacco mosaic virus replicase-mediated cross-protection: contributions of RNA and protein-derived mechanisms.

Specific sequences of the tobacco mosaic virus (TMV) RNA-dependent RNA-polymerase (RdRp) gene were investigated for their ability to confer cross-protection. Nine overlapping segments ranging from 713 to 1070 nucleotides in length and covering the methyltransferase, helicase, and polymerase (POL) domains of the TMV RdRp open reading frame were systemically expressed in Nicotiana benthamiana using a potato X virus (PVX) vector [Chapman, S., Kavanagh, T., and Baulcombe, D. C. (1992). Plant J., 1, 549-557]. PVX-infected plants were subsequently challenge inoculated with 10 microg of wild-type TMV and monitored for TMV accumulation. Mock inoculated plants and plants preinfected with the unmodified PVX vector rapidly accumulated high levels of challenge virus. In contrast, plants preinfected with PVX vectors expressing segments of the TMV RdRp open reading frame displayed either high or low levels of protection. High protection levels were observed for PVX constructs expressing segments of the TMV POL domain, whereas low protection levels were observed for PVX constructs expressing segments covering the methyltransferase and helicase domains. Frameshift mutations that blocked protein expression from RdRp segments disrupted only the high levels of protection derived from POL segments and not the low levels derived from the other segments. However, all RdRp segments conferred similarly high levels of protection against a TMV construct with restricted local movement. Thus both RNA and protein sequences in conjunction with the speed of the infecting challenge virus can affect the protection derived from the TMV RdRp gene.