The NAM, ATAF1/2, and CUC2 (NAC) transcription factors form a large plant-specific gene family, which is involved in the regulation of tissue development in response to biotic and abiotic stress. To date, there have been no comprehensive studies investigating chromosomal location, gene structure, gene phylogeny, conserved motifs, or gene expression of NAC in pepper (Capsicum annuum L.). The recent release of the complete genome sequence of pepper allowed us to perform a genome-wide investigation of Capsicum annuum L. NAC (CaNAC) proteins. In the present study, a comprehensive analysis of the CaNAC gene family in pepper was performed, and a total of 104 CaNAC genes were identified. Genome mapping analysis revealed that CaNAC genes were enriched on four chromosomes (chromosomes 1, 2, 3, and 6). In addition, phylogenetic analysis of the NAC domains from pepper, potato, Arabidopsis, and rice showed that CaNAC genes could be clustered into three groups (I, II, and III). Group III, which contained 24 CaNAC genes, was exclusive to the Solanaceae plant family. Gene structure and protein motif analyses showed that these genes were relatively conserved within each subgroup. The number of introns in CaNAC genes varied from 0 to 8, with 83 (78.9%) of CaNAC genes containing two or less introns. Promoter analysis confirmed that CaNAC genes are involved in pepper growth, development, and biotic or abiotic stress responses. Further, the expression of 22 selected CaNAC genes in response to seven different biotic and abiotic stresses [salt, heat shock, drought, Phytophthora capsici, abscisic acid, salicylic acid (SA), and methyl jasmonate (MeJA)] was evaluated by quantitative RT-PCR to determine their stress-related expression patterns. Several putative stress-responsive CaNAC genes, including CaNAC72 and CaNAC27, which are orthologs of the known stress-responsive Arabidopsis gene ANAC055 and potato gene StNAC30, respectively, were highly regulated by treatment with different types of stress. Our results also showed that CaNAC36 plays an important role in the interaction network, interacting with 48 genes. Most of these genes are in the mitogen-activated protein kinase (MAPK) family. Taken together, our results provide a platform for further studies to identify the biological functions of CaNAC genes.

The presence of 7-epi zingiberene in wild tomatoes has been associated with arthropod resistance. Consequently, tomato breeders are attempting to introgress 7-epi zingiberene from wild to cultivated tomato requiring quantification of zingiberene. 7-Epi zingiberene likely absorbs UV light due to its conjugated double bonds and others have claimed that measurement of absorbance at 270 nm of tomato leaflet washes can be used to quantify zingiberene. However, this claim has never been critically evaluated. We initially evaluated this claim in an interspecific hybrid tomato generation that was segregating widely for zingiberene. Results indicated that the method does not obey the Beer–Lambert law. Consequently, we examined in detail aspects of the UV-absorbance of isolated zingiberenoids and leaflet washes obtained from parents and interspecific generations that were segregating for 7-epi zingiberene. Results indicated that isolated zingiberenoids, as well as leaflet washes containing zingiberenoids, have broad absorbance spectra with a λ_max of 264 nm. For isolated zingiberenoids, the relationship between abundance and absorbance at 264 nm did obey the Beer–Lambert law. Average absorbance spectra for leaflet washes from interspecific generation plants showed subtle λ_max shifts. Furthermore, the relationship between absorbance at 264 nm and zingiberenoid content of these generations did not obey the Beer–Lambert law. The use of multiple wavelengths for estimation of zingiberenoids in these breeding generations was explored and the inclusion of additional absorbances at one or two wavelengths always improved estimates. However, identified wavelength(s) differed among generations. Taken together, the results indicate that measurement of absorbance of tomato leaflet washes at a single wavelength is not a reliable quantitative estimate of zingiberenoids in leaflet washes. Estimates can be improved by utilizing absorbance at multiple wavelengths, but the particular wavelengths will vary among generations. Lastly, measurement of absorbance may be useful for identifying those relatively rare individuals in a generation that is widely segregating for zingiberenoid content. However, even in this situation, the determination of the actual 7-epi zingiberene content would need to be backstopped by a valid quantitative method.