ABSTRACT
As a consequence of climate change, heat waves in combination with extended drought periods will be an increasing threat to crop yield. Therefore, breeding stress tolerant crop plants is an urgent need. Breeding for stress tolerance has benefited from large scale phenotyping, enabling non-invasive, continuous monitoring of plant growth. In case of potato, this is compromised by the fact that tubers grow belowground, making phenotyping of tuber development a challenging task. To determine the growth dynamics of tubers before, during and after stress treatment is nearly impossible with traditional destructive harvesting approaches. In contrast, X-ray Computed Tomography (CT) offers the opportunity to access belowground growth processes. In this study, potato tuber development from initiation until harvest was monitored by CT analysis for five different genotypes under stress conditions. Tuber growth was monitored three times per week via CT analysis. Stress treatment was started when all plants exhibited detectable tubers. Combined heat and drought stress was applied by increasing growth temperature for 2 weeks and simultaneously decreasing daily water supply. CT analysis revealed that tuber growth is inhibited under stress within a week and can resume after the stress has been terminated. After cessation of stress, tubers started growing again and were only slightly and insignificantly smaller than control tubers at the end of the experimental period. These growth characteristics were accompanied by corresponding changes in gene expression and activity of enzymes relevant for starch metabolism which is the driving force for tuber growth. Gene expression and activity of Sucrose Synthase (SuSy) reaffirmed the detrimental impact of the stress on starch biosynthesis. Perception of the stress treatment by the tubers was confirmed by gene expression analysis of potential stress marker genes whose applicability for potato tubers is further discussed. We established a semi-automatic imaging pipeline to analyze potato tuber delevopment in a medium thoughput (5 min per pot). The imaging pipeline presented here can be scaled up to be used in high-throughput phenotyping systems. However, the combination with automated data processing is the key to generate objective data accelerating breeding efforts to improve abiotic stress tolerance of potato genotypes.
ABSTRACT
BACKGROUND: Starch is the principle constituent of potato tubers and is of considerable importance for food and non-food applications. Its metabolism has been subject of extensive research over the past decades. Despite its importance, a description of the complete inventory of genes involved in starch metabolism and their genome organization in potato plants is still missing. Moreover, mechanisms regulating the expression of starch genes in leaves and tubers remain elusive with regard to differences between transitory and storage starch metabolism, respectively. This study aimed at identifying and mapping the complete set of potato starch genes, and to study their expression pattern in leaves and tubers using different sets of transcriptome data. Moreover, we wanted to uncover transcription factors co-regulated with starch accumulation in tubers in order to get insight into the regulation of starch metabolism. RESULTS: We identified 77 genomic loci encoding enzymes involved in starch metabolism. Novel isoforms of many enzymes were found. Their analysis will help to elucidate mechanisms of starch biosynthesis and degradation. Expression analysis of starch genes led to the identification of tissue-specific isoenzymes suggesting differences in the transcriptional regulation of starch metabolism between potato leaf and tuber tissues. Selection of genes predominantly expressed in developing potato tubers and exhibiting an expression pattern indicative for a role in starch biosynthesis enabled the identification of possible transcriptional regulators of tuber starch biosynthesis by co-expression analysis. CONCLUSIONS: This study provides the annotation of the complete set of starch metabolic genes in potato plants and their genomic localizations. Novel, so far undescribed, enzyme isoforms were revealed. Comparative transcriptome analysis enabled the identification of tuber- and leaf-specific isoforms of starch genes. This finding suggests distinct regulatory mechanisms in transitory and storage starch metabolism. Putative regulatory proteins of starch biosynthesis in potato tubers have been identified by co-expression and their expression was verified by quantitative RT-PCR.
