Limits...
Comparative metabolic and transcriptional analysis of a doubled diploid and its diploid citrus rootstock (C. junos cv. Ziyang xiangcheng) suggests its potential value for stress resistance improvement.

Tan FQ, Tu H, Liang WJ, Long JM, Wu XM, Zhang HY, Guo WW - BMC Plant Biol. (2015)

Bottom Line: By RNA-seq analysis, only 212 genes (0.8% of detected genes) are found significantly differentially expressed between 2x and 4x leaves.However, the effect of tetraploidization on transcriptome is limited.Compared to diploid, higher expression level of stress related genes and higher content of stress related metabolites in doubled diploid could be beneficial for its stress tolerance.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Polyploidy has often been considered to confer plants a better adaptation to environmental stresses. Tetraploid citrus rootstocks are expected to have stronger stress tolerance than diploid. Plenty of doubled diploid citrus plants were exploited from diploid species for citrus rootstock improvement. However, limited metabolic and molecular information related to tetraploidization is currently available at a systemic biological level. This study aimed to evaluate the occurrence and extent of metabolic and transcriptional changes induced by tetraploidization in Ziyang xiangcheng (Citrus junos Sieb. ex Tanaka), which is a special citrus germplasm native to China and widely used as an iron deficiency tolerant citrus rootstock.

Results: Doubled diploid Ziyang xiangcheng has typical morphological and anatomical features such as shorter plant height, larger and thicker leaves, bigger stomata and lower stomatal density, compared to its diploid parent. GC-MS (Gas chromatography coupled to mass spectrometry) analysis revealed that tetraploidization has an activation effect on the accumulation of primary metabolites in leaves; many stress-related metabolites such as sucrose, proline and γ-aminobutyric acid (GABA) was remarkably up-regulated in doubled diploid. However, LC-QTOF-MS (Liquid chromatography quadrupole time-of-flight mass spectrometry) analysis demonstrated that tetraploidization has an inhibition effect on the accumulation of secondary metabolites in leaves; all the 33 flavones were down-regulated while all the 6 flavanones were up-regulated in 4x. By RNA-seq analysis, only 212 genes (0.8% of detected genes) are found significantly differentially expressed between 2x and 4x leaves. Notably, those genes were highly related to stress-response functions, including responses to salt stress, water and abscisic acid. Interestingly, the transcriptional divergence could not explain the metabolic changes, probably due to post-transcriptional regulation.

Conclusion: Taken together, tetraploidization induced considerable changes in leaf primary and secondary metabolite accumulation in Ziyang xiangcheng. However, the effect of tetraploidization on transcriptome is limited. Compared to diploid, higher expression level of stress related genes and higher content of stress related metabolites in doubled diploid could be beneficial for its stress tolerance.

Show MeSH

Related in: MedlinePlus

Expression analysis of top 10 up-regulated functionally characterized DEGs in 4× Ziyang xiangcheng by qPCR.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4374211&req=5

Fig6: Expression analysis of top 10 up-regulated functionally characterized DEGs in 4× Ziyang xiangcheng by qPCR.

Mentions: To validate the RNA-seq data, the following top 10 up-regulated functionally characterized genes were selected for qPCR assays: Fe(II)/ascorbate oxidase (SRG1, Cs9g09290), UDP-glucoronosyl/UDP-glucosyltransferase family protein (UGT, Cs5g11620), myb family transcription factor (RL6, Cs3g24870), caffeic acid O-methyltransferase (COMT, orange1.1 t02085), aminocyclopropane 1-carboxylic acid oxidase (ACO, Cs9g08990), u-box armadillo repeat protein (PUB19, Cs7g08470), ethylene response factor (ERF4, Cs1g07950), tracheary element vacuolar protein (XCP1, Cs2g27860), glycosyltransferase (GATL9, Cs7g07900), ethylene response factor (ERF9, Cs2g05620) (Additional file 9). As shown in Figure 6, all the 10 genes were verified to be up-regulated by qPCR analysis, although their fold changes differed from the result of RNA-seq. Notably, six of these genes, namely, SRG1 [39], COMT [40], ACO [41], PUB19 [42], ERF4 [43] and ERF9 [44] were involved in abiotic stress response.Figure 6


Comparative metabolic and transcriptional analysis of a doubled diploid and its diploid citrus rootstock (C. junos cv. Ziyang xiangcheng) suggests its potential value for stress resistance improvement.

Tan FQ, Tu H, Liang WJ, Long JM, Wu XM, Zhang HY, Guo WW - BMC Plant Biol. (2015)

Expression analysis of top 10 up-regulated functionally characterized DEGs in 4× Ziyang xiangcheng by qPCR.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4374211&req=5

Fig6: Expression analysis of top 10 up-regulated functionally characterized DEGs in 4× Ziyang xiangcheng by qPCR.
Mentions: To validate the RNA-seq data, the following top 10 up-regulated functionally characterized genes were selected for qPCR assays: Fe(II)/ascorbate oxidase (SRG1, Cs9g09290), UDP-glucoronosyl/UDP-glucosyltransferase family protein (UGT, Cs5g11620), myb family transcription factor (RL6, Cs3g24870), caffeic acid O-methyltransferase (COMT, orange1.1 t02085), aminocyclopropane 1-carboxylic acid oxidase (ACO, Cs9g08990), u-box armadillo repeat protein (PUB19, Cs7g08470), ethylene response factor (ERF4, Cs1g07950), tracheary element vacuolar protein (XCP1, Cs2g27860), glycosyltransferase (GATL9, Cs7g07900), ethylene response factor (ERF9, Cs2g05620) (Additional file 9). As shown in Figure 6, all the 10 genes were verified to be up-regulated by qPCR analysis, although their fold changes differed from the result of RNA-seq. Notably, six of these genes, namely, SRG1 [39], COMT [40], ACO [41], PUB19 [42], ERF4 [43] and ERF9 [44] were involved in abiotic stress response.Figure 6

Bottom Line: By RNA-seq analysis, only 212 genes (0.8% of detected genes) are found significantly differentially expressed between 2x and 4x leaves.However, the effect of tetraploidization on transcriptome is limited.Compared to diploid, higher expression level of stress related genes and higher content of stress related metabolites in doubled diploid could be beneficial for its stress tolerance.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Polyploidy has often been considered to confer plants a better adaptation to environmental stresses. Tetraploid citrus rootstocks are expected to have stronger stress tolerance than diploid. Plenty of doubled diploid citrus plants were exploited from diploid species for citrus rootstock improvement. However, limited metabolic and molecular information related to tetraploidization is currently available at a systemic biological level. This study aimed to evaluate the occurrence and extent of metabolic and transcriptional changes induced by tetraploidization in Ziyang xiangcheng (Citrus junos Sieb. ex Tanaka), which is a special citrus germplasm native to China and widely used as an iron deficiency tolerant citrus rootstock.

Results: Doubled diploid Ziyang xiangcheng has typical morphological and anatomical features such as shorter plant height, larger and thicker leaves, bigger stomata and lower stomatal density, compared to its diploid parent. GC-MS (Gas chromatography coupled to mass spectrometry) analysis revealed that tetraploidization has an activation effect on the accumulation of primary metabolites in leaves; many stress-related metabolites such as sucrose, proline and γ-aminobutyric acid (GABA) was remarkably up-regulated in doubled diploid. However, LC-QTOF-MS (Liquid chromatography quadrupole time-of-flight mass spectrometry) analysis demonstrated that tetraploidization has an inhibition effect on the accumulation of secondary metabolites in leaves; all the 33 flavones were down-regulated while all the 6 flavanones were up-regulated in 4x. By RNA-seq analysis, only 212 genes (0.8% of detected genes) are found significantly differentially expressed between 2x and 4x leaves. Notably, those genes were highly related to stress-response functions, including responses to salt stress, water and abscisic acid. Interestingly, the transcriptional divergence could not explain the metabolic changes, probably due to post-transcriptional regulation.

Conclusion: Taken together, tetraploidization induced considerable changes in leaf primary and secondary metabolite accumulation in Ziyang xiangcheng. However, the effect of tetraploidization on transcriptome is limited. Compared to diploid, higher expression level of stress related genes and higher content of stress related metabolites in doubled diploid could be beneficial for its stress tolerance.

Show MeSH
Related in: MedlinePlus