Transcriptome analysis of North American sweet birch (Betula lenta) revealed a higher expression of genes involved in the biosynthesis of secondary metabolites than European silver birch (B. pendula).

The North American Betula lenta L. (sweet birch) has been used for medicinal reasons for centuries by native Americans. Although sophisticated technologies have rapidly been developed, a large information gap has been observed regarding genetic regulators of medicinally important compounds in sweet birch. Very little is known on the different genes involved in secondary metabolic biosynthesis in sweet birch. To gain a deeper insight into genetic factors, we performed a transcriptome analysis of each three biological samples from different independent trees of sweet and European silver birch (B. pendula Roth). This allowed us to precisely quantify the transcripts of about 24,000 expressed genes including 29 prominent candidate genes putatively involved in the biosynthesis of secondary metabolites like terpenoids, and aromatic benzoic acids. A total number of 597 genes were differentially expressed between B. lenta and B. pendula, while 264 and 210 genes showed upregulation in the bark and leaf of B. lenta, respectively. Moreover, we identified 39 transcriptional regulatory elements, involved in secondary metabolite biosynthesis, upregulated in B. lenta. Our study demonstrated the potential of RNA sequencing to identify candidate genes interacting in secondary metabolite biosynthesis in sweet birch. The candidate genes identified in this study could be subjected to genetic engineering to functionally characterize them in sweet birch. This knowledge can be beneficial to the increase of therapeutically important compounds.

Identification and analysis of key genes involved in methyl salicylate biosynthesis in different birch species.

Species of the perennial woody plant genus Betula dominate subalpine forests and play a significant role in preserving biological diversity. In addition to their conventional benefits, birches synthesize a wide range of secondary metabolites having pharmacological significance. Methyl salicylate (MeSA) is one of these naturally occurring compounds constitutively produced by different birch species. MeSA is therapeutically important in human medicine for muscle injuries and joint pain. However, MeSA is now mainly produced synthetically due to a lack of information relating to MeSA biosynthesis and regulation. In this study, we performed a comprehensive bioinformatics analysis of two candidate genes mediating MeSA biosynthesis, SALICYLIC ACID METHYLTRANSFERASE (SAMT) and SALICYLIC ACID-BINDING PROTEIN 2 (SABP2), of high (B. lenta, B. alleghaniensis, B. medwediewii, and B. grossa) and low (B. pendula, B. utilis, B. alnoides, and B. nana) MeSA-producing birch species. Phylogenetic analyses of SAMT and SABP2 genes and homologous genes from other plant species confirmed their evolutionary relationships. Multiple sequence alignments of the amino acid revealed the occurrence of important residues for substrate specificity in SAMT and SABP2. The analysis of cis elements in different birches indicated a functional multiplicity of SAMT and SABP2 and provided insights into the regulation of both genes. We successfully developed six prominent single nucleotide substitution markers that were validated with 38 additional birch individuals to differentiate high and low MeSA-producing birch species. Relative tissue-specific expression analysis of SAMT in leaf and bark tissue of two high and two low MeSA-synthesizing birches revealed a high expression in the bark of both high MeSA-synthesizing birches. In contrast, SABP2 expression in tissues revealed indifferent levels of expression between species belonging to the two groups. The comparative expression and bioinformatics analyses provided vital information that could be used to apply plant genetic engineering technology in the mass production of organic MeSA.

Use of near infrared spectroscopy in online-monitoring of feeding substrate quality in anaerobic digestion.

In order to keep the anaerobic process stably and uniformly producing biogas it needs to be supplied with either an even amount of substrate of stable quality or varying amounts according to variations in quality. Feeding amounts are usually adjusted manually as a reaction to changing rates of biogas production. Continuous information about the actual substrate quality is not available and feedstuff analyses are costly. Aim of this study was to assess the feasibility of near infrared spectroscopic (NIRS) online monitoring of substrate quality in order to find ways towards more exact control of biogas plant feeding. A NIRS sensor system was designed, constructed and calibrated for continuous monitoring of (RMSECV in brackets) dry matter (DM) (0.75%fresh matter (FM)), volatile solids (0.74%FM), crude fat (0.09%FM), crude protein (0.22%FM), crude fiber (1.50%DM) and nitrogen-free extracts (0.93%FM) of maize silage.

Use of near infrared spectroscopy in monitoring of volatile fatty acids in anaerobic digestion.

Recently biogas production from agricultural sources has rapidly developed. Therefore the demands on biogas plants to optimise the efficiency of the anaerobic digestion (AD) process have grown immensely. At present there is no online-supervision tool available to monitor the AD process, but costly and time-consuming chemical analyses are necessary. The possibility to use near-infrared spectroscopy (NIRS) in order to track relevant process parameters like total volatile fatty acids (VFA), acetic acid and propionic acid was investigated in the present research project. A NIR-sensor was integrated into a full-scale 1 MW biogas plant and NIR-spectra of the fermenter contents were recorded semi-continuously for 500 days. Weekly samples were taken and analysed for the above mentioned parameters. Calibration models were calculated, capable of following these parameters: VFA (r(2)=0.94), acetic acid (r(2)=0.69), propionic acid (r(2)=0.89).