Differential expression of heat shock protein genes in sorghum (Sorghum bicolor L.) genotypes under heat stress

Abstract Various types of sorghum were subjected to thermal stress to reveal the mode of expression of genes of the heat shock protein (hsp) family. In silico sequence determination of hsp genes in related cereal species led to the selection of appropriate primers for PCR amplification of a segment corresponding to the hsp90 gene from sorghum. Deduced sequence information allowed the design of gene specific primers for quantification of hsp90 gene expression by means of real-time quantitative polymerase chain reaction (RTqPCR). Fourteen days-old plants were exposed to a temperature of 47°C for a time period ranging from 10 to 180 min. Total RNA was extracted from stressed and control plants and subjected to reverse transcription and RT-qPCR analysis. The actin gene was used as an internal standard. Gene expression was assessed by using cDNA from all types of plant material and for all the different durations of heat stress exposure. Data from RT-qPCR analyses were analyzed using REST software. The highest level of hsp90 gene expression was realized upon exposure to heat for either 60 or 30 min, while expression levels differed among the genotypes studied. In addition, overall levels of hsp90 gene expression were significantly different among varieties tested. Information of such genotypic variation in expression levels of hsp90 gene under heat stress, coupled with related field performance data, could potentially be exploited in breeding programs

Nicotiana benthamiana, a popular model for genome evolution and plant–pathogen interactions

Nicotiana benthamiana originates from northern Australia and belongs to the Suaveolentes section. It is used extensively as a model organism for many types of research, including plant–pathogen interactions, RNA interference, and functional genomics. Recent publications that used N. benthamiana as a model for plant–pathogen interactions focused mainly on bacteria, viruses, oomycete, and fungi. Two different N. benthamiana whole genome assemblies were published in 2012. These assemblies have been improved and structurally annotated in later versions but are still incomplete. The lineage most widely used in research originates from a population that has retained a loss-of-function mutation in Rdr1 (RNA-dependent RNA polymerase 1) that makes it highly susceptible to viruses. In this chapter, we review some of the techniques used in N. benthamiana to study plant–pathogen interactions, including virus-induced gene silencing, transient protein expression by agroinfiltration, stable genetic manipulation, and transcriptomics analysis, and discuss some of the results. Descriptions and links to some of the most relevant online resources for N. benthamiana are also provided.Fil: Pombo, Marina Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Rosli, Hernan Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Fernandez Pozo, Noé. University of Marburg; AlemaniaFil: Aureliano, Bombarely. Virginia Polytechnic Institute; Estados Unidos. Università degli Studi di Milano; Itali