Cytological and Transcriptomic Analysis Provide Insights into the Formation of Variegated Leaves in Ilex × altaclerensis ‘Belgica Aurea’
Abstract
:1. Introduction
2. Results
2.1. Leaf Anatomical Characteristics and Ultrastructure
2.2. Pigment Content in Yellow and Variegated Leaves
2.3. Localization of Pigments in the Leaves
2.4. RNA-Seq Analysis
2.5. Gene Annotation and Function Classification
2.6. Gene Expression Analysis
2.7. GO Enrichment Analysis of DEGs
2.8. KEGG Enrichment Analysis of DEGs
2.9. Analysis of the Genes Involved in Chlorophyll Metabolic Pathway and Chloroplast Integrity and Function
2.10. Validation of RNA Sequencing Data by qRT-PCR
3. Discussion
4. Materials and Methods
4.1. Plant Materials
4.2. Measurements of Contents of Chlorophyll, Carotenoids and Anthocyanins
4.3. Transmission Electron Microscopic Observation
4.4. Leaf Histology
4.5. Laser Scanning Confocal Microscopy
4.6. RNA Extraction and cDNA Library Preparation
4.7. Transcriptome Assembly and Functional Annotation
4.8. Analysis of Differentially Expressed Genes (DEGs)
4.9. Quantitative Real-Time PCR
4.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Yellow | Green | Variegation | |
---|---|---|---|
Chl a (mg·g−1 FW) | 0.09 ± 0.01 b | 1.28 ± 0.09 a | 0.11 ± 0.01 b |
Chl b (mg·g−1 FW) | 0.05 ± 0.01 c | 0.38 ± 0.01 a | 0.10 ± 0.01 b |
Chl a+b (mg·g−1 FW) | 0.14 ± 0.01 b | 1.66 ± 0.10 a | 0.21 ± 0.01 b |
Chl a/b | 1.63 ± 0.07 b | 3.33 ± 0.12 a | 1.14 ± 0.03 c |
Car (mg·g−1 FW) | 0.05 ± 0.01 b | 0.46 ± 0.03 a | 0.06 ± 0.01 b |
Chl a+b/Car | 2.91 ± 0.12 c | 3.65 ± 0.04 a | 3.31 ± 0.09 b |
Ant (mg·g−1 FW) | 0.10 ± 0.01 b | 0.145 ± 0.01 a | 0.10 ± 0.01 b |
Database | Number of Annotated Unigenes | Percentage of Annotated Unigenes |
---|---|---|
NR | 59,752 | 75.98% |
NT | 52,105 | 66.26% |
Swiss-Prot | 45,066 | 57.31% |
KEGG | 47,967 | 61.00% |
KOG | 47,369 | 60.24% |
Pfam | 46,101 | 58.63% |
GO | 34,793 | 44.25% |
Intersection | 20,462 | 26.02% |
Overall | 62,275 | 79.19% |
Gene ID | Log2 Fold Change | Annotation | ||
---|---|---|---|---|
Variegation vs. Green | Yellow vs. Green | Yellow vs. Variegation | ||
CL5332.Contig4_All | 3.20 | 2.16 | −1.04 | magnesium-protoporphyrin IX monomethyl ester [oxidative] cyclase |
CL374.Contig9_All | 8.64 | 3.97 | −4.66 | chlorophyll(ide) b reductase |
CL8272.Contig2_All | 7.34 | 2.73 | −4.61 | chlorophyll(ide) b reductase |
CL8272.Contig3_All | −3.41 | −2.15 | 1.26 | chlorophyll(ide) b reductase |
Unigene31820_All | 8.64 | 2.79 | −5.85 | monogalactosyldiacylglycerol synthase |
CL3869.Contig6_All | 2.09 | 8.83 | 6.73 | digalactosyldiacylglycerol synthase |
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Zhang, Q.; Huang, J.; Zhou, P.; Hao, M.; Zhang, M. Cytological and Transcriptomic Analysis Provide Insights into the Formation of Variegated Leaves in Ilex × altaclerensis ‘Belgica Aurea’. Plants 2021, 10, 552. https://doi.org/10.3390/plants10030552
Zhang Q, Huang J, Zhou P, Hao M, Zhang M. Cytological and Transcriptomic Analysis Provide Insights into the Formation of Variegated Leaves in Ilex × altaclerensis ‘Belgica Aurea’. Plants. 2021; 10(3):552. https://doi.org/10.3390/plants10030552
Chicago/Turabian StyleZhang, Qiang, Jing Huang, Peng Zhou, Mingzhuo Hao, and Min Zhang. 2021. "Cytological and Transcriptomic Analysis Provide Insights into the Formation of Variegated Leaves in Ilex × altaclerensis ‘Belgica Aurea’" Plants 10, no. 3: 552. https://doi.org/10.3390/plants10030552