Limits...
Use of the de novo transcriptome analysis of silver-leaf nightshade (Solanum elaeagnifolium) to identify gene expression changes associated with wounding and terpene biosynthesis.

Tsaballa A, Nikolaidis A, Trikka F, Ignea C, Kampranis SC, Makris AM, Argiriou A - BMC Genomics (2015)

Bottom Line: Analysis of wounded S. elaeagnifolium leaves has shown significant increase of the concentration of (E)-caryophyllene and geranyl linalool, two terpenes implicated in stress responses.The increased production of (E)-caryophyllene was matched to the induced expression of the corresponding TPS gene.Analysis of genes and pathways involved in the plant's interaction with the environment will help to elucidate the mechanisms that underly the intricate features of this unique Solanum species.

View Article: PubMed Central - PubMed

Affiliation: Institute of Applied Biosciences, Center for Research and Technology Hellas (CERTH), P.O. Box 60361, Thessaloniki, 57001, Greece. afroditi.tsampalla@gmail.com.

ABSTRACT

Background: Solanum elaeagnifolium, an invasive weed of the Solanaceae family, is poorly studied although it poses a significant threat to crops. Here the analysis of the transcriptome of S. elaeagnifolium is presented, as a means to explore the biology of this species and to identify genes related to its adaptation to environmental stress. One of the basic mechanisms by which plants respond to environmental stress is through the synthesis of specific secondary metabolites that protect the plant from herbivores and microorganisms, or serve as signaling molecules. One important such group of secondary metabolites are terpenes.

Results: By next-generation sequencing, the flower/leaf transcriptome of S. elaeagnifolium was sequenced and de novo assembled into 75,618 unigenes. Among the unigenes identified, several corresponded to genes involved in terpene biosynthesis; these included terpene synthases (TPSs) and genes of the mevalonate (MVA) and the methylerythritol phosphate (MEP) pathways. Functional characterization of two of the TPSs showed that one produced the sesquiterpene (E)-caryophyllene and the second produced the monoterpene camphene. Analysis of wounded S. elaeagnifolium leaves has shown significant increase of the concentration of (E)-caryophyllene and geranyl linalool, two terpenes implicated in stress responses. The increased production of (E)-caryophyllene was matched to the induced expression of the corresponding TPS gene. Wounding also led to the increased expression of the putative 1-deoxy-D-xylulose-5-phosphate synthase 2 (DXS2) gene, a key enzyme of the MEP pathway, corroborating the overall increased output of terpene biosynthesis.

Conclusions: The reported S. elaeagnifolium de novo transcriptome provides a valuable sequence database that could facilitate study of this invasive weed and contribute to our understanding of the highly diverse Solanaceae family. Analysis of genes and pathways involved in the plant's interaction with the environment will help to elucidate the mechanisms that underly the intricate features of this unique Solanum species.

No MeSH data available.


Related in: MedlinePlus

GC/MS chromatograms. a GC/MS chromatogram with the substances produced by cl9841 expressed in yeast. b GC/MS chromatogram of the dodecane phase of yeast culture. The major product seems to be caryophyllene (26.26 min) as its peak area percentage reflexes a higher amount compared to α-humulene (27.23 min) which is also produced by yeast
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4492009&req=5

Fig3: GC/MS chromatograms. a GC/MS chromatogram with the substances produced by cl9841 expressed in yeast. b GC/MS chromatogram of the dodecane phase of yeast culture. The major product seems to be caryophyllene (26.26 min) as its peak area percentage reflexes a higher amount compared to α-humulene (27.23 min) which is also produced by yeast

Mentions: The yeast strain AM94 [31] was used to transform cl9841 putative monoterpene synthase together with the ERG20 (F96W-N127W) variant which shifts production towards GPP substrate [32]. For the expression of cl1310, the gene was co-expressed in AM238 cells together with copalyl diphosphate synthase from Salvia pomifera and a variant of yeast ERG20 (F96C) producing GGPP. For the characterization of the putative sesquiterpene cl7653, the yeast strain AM109 was used [31]. The cl7653 carrying plasmid was transformed either alone or together with a stabilised variant of HMG2(K6R) to increase substrate availability [33]. As seen in Fig. 3a, cl9841 is an active monoterpene synthase enzyme producing a range of monoterpenes with the most prominent being camphene (52.55 %), β-myrcene (11.01 %) and limonene (10.44 %) and several minor additional compounds. The cl1310 expressing cells did not produce any compounds. The cl7653 enzyme was active and less promiscuous than cl9841, producing mainly caryophyllene (86.4 %) and lesser amounts of α-humulene (Fig. 3b). The caryophyllene peak was additionally validated with the mass spectrum of a standard compound.Fig. 3


Use of the de novo transcriptome analysis of silver-leaf nightshade (Solanum elaeagnifolium) to identify gene expression changes associated with wounding and terpene biosynthesis.

Tsaballa A, Nikolaidis A, Trikka F, Ignea C, Kampranis SC, Makris AM, Argiriou A - BMC Genomics (2015)

GC/MS chromatograms. a GC/MS chromatogram with the substances produced by cl9841 expressed in yeast. b GC/MS chromatogram of the dodecane phase of yeast culture. The major product seems to be caryophyllene (26.26 min) as its peak area percentage reflexes a higher amount compared to α-humulene (27.23 min) which is also produced by yeast
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4492009&req=5

Fig3: GC/MS chromatograms. a GC/MS chromatogram with the substances produced by cl9841 expressed in yeast. b GC/MS chromatogram of the dodecane phase of yeast culture. The major product seems to be caryophyllene (26.26 min) as its peak area percentage reflexes a higher amount compared to α-humulene (27.23 min) which is also produced by yeast
Mentions: The yeast strain AM94 [31] was used to transform cl9841 putative monoterpene synthase together with the ERG20 (F96W-N127W) variant which shifts production towards GPP substrate [32]. For the expression of cl1310, the gene was co-expressed in AM238 cells together with copalyl diphosphate synthase from Salvia pomifera and a variant of yeast ERG20 (F96C) producing GGPP. For the characterization of the putative sesquiterpene cl7653, the yeast strain AM109 was used [31]. The cl7653 carrying plasmid was transformed either alone or together with a stabilised variant of HMG2(K6R) to increase substrate availability [33]. As seen in Fig. 3a, cl9841 is an active monoterpene synthase enzyme producing a range of monoterpenes with the most prominent being camphene (52.55 %), β-myrcene (11.01 %) and limonene (10.44 %) and several minor additional compounds. The cl1310 expressing cells did not produce any compounds. The cl7653 enzyme was active and less promiscuous than cl9841, producing mainly caryophyllene (86.4 %) and lesser amounts of α-humulene (Fig. 3b). The caryophyllene peak was additionally validated with the mass spectrum of a standard compound.Fig. 3

Bottom Line: Analysis of wounded S. elaeagnifolium leaves has shown significant increase of the concentration of (E)-caryophyllene and geranyl linalool, two terpenes implicated in stress responses.The increased production of (E)-caryophyllene was matched to the induced expression of the corresponding TPS gene.Analysis of genes and pathways involved in the plant's interaction with the environment will help to elucidate the mechanisms that underly the intricate features of this unique Solanum species.

View Article: PubMed Central - PubMed

Affiliation: Institute of Applied Biosciences, Center for Research and Technology Hellas (CERTH), P.O. Box 60361, Thessaloniki, 57001, Greece. afroditi.tsampalla@gmail.com.

ABSTRACT

Background: Solanum elaeagnifolium, an invasive weed of the Solanaceae family, is poorly studied although it poses a significant threat to crops. Here the analysis of the transcriptome of S. elaeagnifolium is presented, as a means to explore the biology of this species and to identify genes related to its adaptation to environmental stress. One of the basic mechanisms by which plants respond to environmental stress is through the synthesis of specific secondary metabolites that protect the plant from herbivores and microorganisms, or serve as signaling molecules. One important such group of secondary metabolites are terpenes.

Results: By next-generation sequencing, the flower/leaf transcriptome of S. elaeagnifolium was sequenced and de novo assembled into 75,618 unigenes. Among the unigenes identified, several corresponded to genes involved in terpene biosynthesis; these included terpene synthases (TPSs) and genes of the mevalonate (MVA) and the methylerythritol phosphate (MEP) pathways. Functional characterization of two of the TPSs showed that one produced the sesquiterpene (E)-caryophyllene and the second produced the monoterpene camphene. Analysis of wounded S. elaeagnifolium leaves has shown significant increase of the concentration of (E)-caryophyllene and geranyl linalool, two terpenes implicated in stress responses. The increased production of (E)-caryophyllene was matched to the induced expression of the corresponding TPS gene. Wounding also led to the increased expression of the putative 1-deoxy-D-xylulose-5-phosphate synthase 2 (DXS2) gene, a key enzyme of the MEP pathway, corroborating the overall increased output of terpene biosynthesis.

Conclusions: The reported S. elaeagnifolium de novo transcriptome provides a valuable sequence database that could facilitate study of this invasive weed and contribute to our understanding of the highly diverse Solanaceae family. Analysis of genes and pathways involved in the plant's interaction with the environment will help to elucidate the mechanisms that underly the intricate features of this unique Solanum species.

No MeSH data available.


Related in: MedlinePlus