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Investigation of the multifunctional gene AOP3 expands the regulatory network fine-tuning glucosinolate production in Arabidopsis.

Jensen LM, Kliebenstein DJ, Burow M - Front Plant Sci (2015)

Bottom Line: In this study, we use transgenic plants in combination with natural variation to investigate the regulatory role of the AOP3 gene found in GS-AOP locus previously suggested to contribute to the regulation of glucosinolate defense compounds.Phenotypic analysis and QTL mapping in F2 populations with different AOP3 transgenes support that the enzymatic function and the AOP3 RNA both play a significant role in controlling glucosinolate accumulation.Furthermore, we find different loci interacting with either the enzymatic activity or the RNA of AOP3 and thereby extend the regulatory network controlling glucosinolate accumulation.

View Article: PubMed Central - PubMed

Affiliation: DNRF Center DynaMo, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen Frederiksberg, Denmark ; Copenhagen Plant Science Centre, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen Frederiksberg, Denmark.

ABSTRACT
Quantitative trait loci (QTL) mapping studies enable identification of loci that are part of regulatory networks controlling various phenotypes. Detailed investigations of genes within these loci are required to ultimately understand the function of individual genes and how they interact with other players in the network. In this study, we use transgenic plants in combination with natural variation to investigate the regulatory role of the AOP3 gene found in GS-AOP locus previously suggested to contribute to the regulation of glucosinolate defense compounds. Phenotypic analysis and QTL mapping in F2 populations with different AOP3 transgenes support that the enzymatic function and the AOP3 RNA both play a significant role in controlling glucosinolate accumulation. Furthermore, we find different loci interacting with either the enzymatic activity or the RNA of AOP3 and thereby extend the regulatory network controlling glucosinolate accumulation.

No MeSH data available.


Related in: MedlinePlus

Allele specific interactions of AOP3 and QTLs controlling different glucosinolates. (A) Average leaf levels of 3msp dependent on the genetic state of the interaction of AOP3 and X122. (B) Levels of 4msb dependent on AOP3 and X122. (C) Accumulation of 8mso depends on the state of AOP3. (D) Levels of I3M controlled by the interaction of AOP3 and X186. “0” indicates homozygous for absence of AOP3 or the Col-0 allele, “1” heterozygous for AOP3 or the marker, and “2” homozygous for presence of AOP3 or the Gie-0 allele.
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Figure 7: Allele specific interactions of AOP3 and QTLs controlling different glucosinolates. (A) Average leaf levels of 3msp dependent on the genetic state of the interaction of AOP3 and X122. (B) Levels of 4msb dependent on AOP3 and X122. (C) Accumulation of 8mso depends on the state of AOP3. (D) Levels of I3M controlled by the interaction of AOP3 and X186. “0” indicates homozygous for absence of AOP3 or the Col-0 allele, “1” heterozygous for AOP3 or the marker, and “2” homozygous for presence of AOP3 or the Gie-0 allele.

Mentions: Based on the results from the mapping in the individual populations, where AOP3 UT showed epistasis with different QTLs in the UT2 and UT10 populations for different glucosinolates (Table 3), we made combined models only including main effect QTLs and epistatic interactions significant in the pooled UT populations (Table S1). This allowed us to test across different insertion sites and other population effects. The analysis revealed that there is a consistently significant interaction of AOP3 UT and a locus near marker X122 on chromosome 2 for controlling 3msp accumulation across the populations (Table 4). Allele-specific analysis showed a semi-dominant effect of the presence of AOP3 and the Col-0 allele for X122 for 3msp (Figure 7). A similar effect is not seen for accumulation of the main C4 glucosinolate, 4-methylsulfinylbutyl glucosinolate (4msb), illustrating the fine-tuning regulatory effect of the AOP3 RNA. Additionally, the AOP3 UT construct shows up as significant for controlling the main LC glucosinolate 8-methylsulfinyloctyl glucosinolate (8mso), in the combined populations. However, in this case, no significant interacting loci were found (Table 4). The highest levels of 8mso is found in plants homozygous for AOP3, suggesting a dose-dependent effect of the RNA. We also tested for the levels of the indole glucosinolate I3M and found that the AOP3 RNA also influences the accumulation by interaction with X186 on chromosome 4 (Table 4). The allele-specific interactions showed a semi-dominant pattern of interaction between the AOP3 RNA and the Col-0 allele of X186.


Investigation of the multifunctional gene AOP3 expands the regulatory network fine-tuning glucosinolate production in Arabidopsis.

Jensen LM, Kliebenstein DJ, Burow M - Front Plant Sci (2015)

Allele specific interactions of AOP3 and QTLs controlling different glucosinolates. (A) Average leaf levels of 3msp dependent on the genetic state of the interaction of AOP3 and X122. (B) Levels of 4msb dependent on AOP3 and X122. (C) Accumulation of 8mso depends on the state of AOP3. (D) Levels of I3M controlled by the interaction of AOP3 and X186. “0” indicates homozygous for absence of AOP3 or the Col-0 allele, “1” heterozygous for AOP3 or the marker, and “2” homozygous for presence of AOP3 or the Gie-0 allele.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4585220&req=5

Figure 7: Allele specific interactions of AOP3 and QTLs controlling different glucosinolates. (A) Average leaf levels of 3msp dependent on the genetic state of the interaction of AOP3 and X122. (B) Levels of 4msb dependent on AOP3 and X122. (C) Accumulation of 8mso depends on the state of AOP3. (D) Levels of I3M controlled by the interaction of AOP3 and X186. “0” indicates homozygous for absence of AOP3 or the Col-0 allele, “1” heterozygous for AOP3 or the marker, and “2” homozygous for presence of AOP3 or the Gie-0 allele.
Mentions: Based on the results from the mapping in the individual populations, where AOP3 UT showed epistasis with different QTLs in the UT2 and UT10 populations for different glucosinolates (Table 3), we made combined models only including main effect QTLs and epistatic interactions significant in the pooled UT populations (Table S1). This allowed us to test across different insertion sites and other population effects. The analysis revealed that there is a consistently significant interaction of AOP3 UT and a locus near marker X122 on chromosome 2 for controlling 3msp accumulation across the populations (Table 4). Allele-specific analysis showed a semi-dominant effect of the presence of AOP3 and the Col-0 allele for X122 for 3msp (Figure 7). A similar effect is not seen for accumulation of the main C4 glucosinolate, 4-methylsulfinylbutyl glucosinolate (4msb), illustrating the fine-tuning regulatory effect of the AOP3 RNA. Additionally, the AOP3 UT construct shows up as significant for controlling the main LC glucosinolate 8-methylsulfinyloctyl glucosinolate (8mso), in the combined populations. However, in this case, no significant interacting loci were found (Table 4). The highest levels of 8mso is found in plants homozygous for AOP3, suggesting a dose-dependent effect of the RNA. We also tested for the levels of the indole glucosinolate I3M and found that the AOP3 RNA also influences the accumulation by interaction with X186 on chromosome 4 (Table 4). The allele-specific interactions showed a semi-dominant pattern of interaction between the AOP3 RNA and the Col-0 allele of X186.

Bottom Line: In this study, we use transgenic plants in combination with natural variation to investigate the regulatory role of the AOP3 gene found in GS-AOP locus previously suggested to contribute to the regulation of glucosinolate defense compounds.Phenotypic analysis and QTL mapping in F2 populations with different AOP3 transgenes support that the enzymatic function and the AOP3 RNA both play a significant role in controlling glucosinolate accumulation.Furthermore, we find different loci interacting with either the enzymatic activity or the RNA of AOP3 and thereby extend the regulatory network controlling glucosinolate accumulation.

View Article: PubMed Central - PubMed

Affiliation: DNRF Center DynaMo, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen Frederiksberg, Denmark ; Copenhagen Plant Science Centre, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen Frederiksberg, Denmark.

ABSTRACT
Quantitative trait loci (QTL) mapping studies enable identification of loci that are part of regulatory networks controlling various phenotypes. Detailed investigations of genes within these loci are required to ultimately understand the function of individual genes and how they interact with other players in the network. In this study, we use transgenic plants in combination with natural variation to investigate the regulatory role of the AOP3 gene found in GS-AOP locus previously suggested to contribute to the regulation of glucosinolate defense compounds. Phenotypic analysis and QTL mapping in F2 populations with different AOP3 transgenes support that the enzymatic function and the AOP3 RNA both play a significant role in controlling glucosinolate accumulation. Furthermore, we find different loci interacting with either the enzymatic activity or the RNA of AOP3 and thereby extend the regulatory network controlling glucosinolate accumulation.

No MeSH data available.


Related in: MedlinePlus