Acquired thermotolerance and expression of the HSP100/ClpB genes of lima bean.

Acquired thermotolerance (AT) is the ability of cells to survive a normally lethal temperature treatment as a consequence of pretreatment at an elevated but sublethal temperature. In yeast and cyanobacteria, the expression of the HSP100/ClpB protein is required for the AT response. To determine whether the HSP100/ClpB protein is associated with this response in lima bean (Phaseolus lunatus), we have cloned an HSP100/ClpB homolog and assessed expression of the two gene copies under heat stress conditions, which induce AT. Transcription of the cytoplasmically localized HSP100/ClpB protein genes is stringently controlled by heat stress in both of the laboratory and field heat stress conditions. From a heat-induced cDNA library, we identified a clone of a putative chloroplast-targeted (cp) HSP100/ClpB protein gene sequence. The cp HSP100/ClpB protein genes are constitutively expressed, but transcript levels increase post-heat stress in laboratory heat stress experiments. In field conditions the genes for the cp HSP100/ClpB are constitutively expressed. Although we were unable to correlate differences in the timing of AT response with the expression or genetic structure of the HSP100/ClpB genes in heat-tolerant or -sensitive varieties of lima bean, we clearly demonstrate the association of expression of HSP100/ClpB proteins with heat response in this species.

Research notes: Comparison of disease susceptibility and resistance in three lines of chickens experimentally infected with infectious laryngotracheitis virus.

The susceptibility of three F1 hybrid lines of chickens to graded doses of infectious laryngotracheitis virus (ILTV) was investigated. The three F1 hybrid lines, each produced from mating two inbred lines, included the SC (B2B2) and TK (B15B21) lines and the 15I5 x 7(1) (B2B15) line. Although at 1 d of age all three lines were susceptible to ILTV, SC birds were significantly less susceptible (10%) than TK (80%) or 15I5 x 7(1) (50%) birds when exposed to 5,000 pfu of virus at 4 wk of age. The ability of each inbred F1 hybrid line to establish a protective immune response to ILTV was also determined. The SC birds required a smaller immunizing dose of virus (500 pfu) to mount a protective immune response to ILTV than the 15I5 x 7(1) line (5,000 pfu). A 5,000 pfu immunizing dose did not elicit a protective immune response in the TK line to a 10(6) pfu challenge dose of ILTV. These results also correlated with the ability to produce ILTV-specific antibodies. This study confirms and expands on observations that lines of chickens differ with respect to their susceptibility and resistance to ILTV.

Comparison of in vitro and in vivo systems for propagation of Rift Valley fever virus from clinical specimens.

Several cell cultures and animals were compared for their relative sensitivity as primary isolation systems for Rift Valley fever virus (RVFV) and to determine if virulence characteristics of the isolates were altered in these systems. Eleven human sera from known cases of Rift Valley fever (RVF) were obtained from the 1987 epidemic in Mauritania and served as the source of virus for these studies. Sera were inoculated directly into cell cultures (Vero, C6/36 and DBS-FRhL-2) and animals (ICR suckling mice, Lak:LVG(SYR) hamsters and WF rats) concurrently. The cell lines provided a quick method to propagate, quantitate and identify these specimens without prior adaption. The isolates were highly virulent for suckling mice and hamsters, but not for WF rats, even after cell culture passage, which indicated that the Mauritanian isolates more closely resembled those strains from sub-Saharan Africa than those from the 1977-78 Egyptian epidemic.