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Differential transcriptome analysis of leaves of tea plant (Camellia sinensis) provides comprehensive insights into the defense responses to Ectropis oblique attack using RNA-Seq

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Abstract

Tea is a very popular and healthy nonalcoholic beverage worldwide. As an evergreen woody plant, the cultivation of tea plants (Camellia sinensis) is challenged by biotic stresses, and one of which is feeding of Ectropis oblique. In China, E. oblique infestation causes serious damages in many tea cultivation areas. Tea plants have evolved sophisticated strategies to cope with attack by E. oblique. To elucidate the molecular mechanisms of the response to E. oblique in tea plants, the differential gene expression profiles between the E. oblique damage-induced tea plants and undamaged control using RNA sequencing (RNA-Seq) were obtained. A total of 1859 differentially expressed genes were identified, including 949 upregulated and 910 downregulated genes. Overall, 90 signal transduction genes, 100 anti-insect responsive transcription factors, 50 genes related to phenylpropanoid biosynthesis, 41 unigenes related to herbivore-induced plant volatiles (HIPVs) biosynthesis, and 8 caffeine biosynthesis genes were found to be differentially regulated. Metabolic pathway analysis indicated that plant secondary metabolites and the signaling pathways may play an important role in defense against insects, and a closer examination at the expression of some crucial genes revealed differential expression patterns after feeding by E. oblique. Furthermore, quantitative RT-PCR (qRT-PCR) analysis further confirmed the results of RNA-Seq. Our dataset provides the most comprehensive sequence resource available for studying the resistance to E. oblique in tea, which will benefit our understanding of the overall mechanisms underlying inducible defenses responses, and may be useful to create novel prevention measures against insects to reduce pesticide usage in eco-friendly tea farming.

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Abbreviations

AACT:

Acetoacetyl CoA thiolase

AAT:

Alcohol acyltransferase

ADH:

Alcohol dehydrogenase

ANR:

Anthocyanidin reductase

bZIP:

Basic-region leucine zipper protein

C:

Catechin

Cab:

Chlorophyll a/b binding protein

CAD:

Cinnamyl alcohol dehydrogenase

CaM:

Calmodulin

cDNA-AFLP:

cDNA-amplified fragment length polymorphisms

CDPKs:

Ca2+-dependent protein kinases

CHS:

Chalcone synthase

COG:

Clusters of orthologous groups of protein

CTAB:

Cetyltrimethyl ammonium bromide

DAHPS:

3-Deoxy-darabinoheptulosonate-7-p-synthase

DDRT-PCR:

Display of reverse transcriptase polymerase chain reaction polymerase chain reaction

DEGs:

Differentially expressed genes

DFR:

Dihydroflavonol-4-reductase

DXR:

1-Deoxy-d-xylulose-5-phosphate-reductoisomerase

EC:

Epicatechin

ECG:

Epicatechin gallate

EGC:

Epigallocatechin

EGCG:

Epigallocatechin gallate

EPPS:

5-Enolpyruvylshikimate 3-phosphate synthase

ET:

Ethylene

FDR:

False discovery rate

FPKM:

Fragments per kilobase of exon model per million mapped reads

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GC:

Gallocatechin

GGPPSase:

Geranylgeranyl diphosphate synthase

GLVs:

Green leaf volatiles

GO:

Gene ontology

HMGR:

3-Hydroxy-3-methylglutaryl-CoA reductase

HMGS:

3-Hydroxy-3-methylglutaryl-CoA synthase

HPL:

Hydroperoxide lyase

IFS:

Isoflavone synthase

HIPVs:

Herbivore induced plant volatiles

JA:

Jasmonic acid

JAZ:

Jasmonate ZIM-domain

KEGG:

Kyoto encyclopedia of genes and genomes

LOX:

Lipoxidase

LRR-RLKs:

Leucine-rich repeat receptor-like kinases

MAPKs:

Mitogen-activated protein kinases

MCS:

2-C-Methyl-d-erythtitol 2,4-cyclodiphosphate synthase

MEP:

2-C-Methyl-d-erythritol 4-phosphate

MVA:

Mevalonate

PAL:

Phenylalanine-ammonia lyase

PMD:

Pyrophosphomevalonate decarboxylase

qRT-PCR:

Quantitative reverse transcriptase Polymerase Chain Reaction

RNA-Seq:

RNA sequencing

ROS:

Reactive oxygen species

SA:

Salicylic acid

SAMS:

S-Adenosylmethionine synthase

SAMT:

Salicylic acid carboxyl methyltransferase

TCS:

Caffeine synthase

TFs:

Transcription factors

TPS:

Terpene synthase

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (NSFC) projects (31171608, 31300578), the Special Innovative Province Construction in Anhui Province (15czs08032), and the Doctoral Science Foundation of Anhui Agricultrual University (wd2016-02) grants. We thank Prof. Jian-Qiang Wu at Kuming Institute of Botany, Chinese Academy of Sciences, China, for discussions and critical reading of the manuscript.

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    1. State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang West Road, Hefei, Anhui, 230036, People’s Republic of China

      Ya-Nan Wang, Lei Tang, Yan Hou, Ping Wang, Hua Yang & Chao-Ling Wei

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    Ya-Nan Wang and Lei Tang contributed equally to this work.

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    Wang, YN., Tang, L., Hou, Y. et al. Differential transcriptome analysis of leaves of tea plant (Camellia sinensis) provides comprehensive insights into the defense responses to Ectropis oblique attack using RNA-Seq. Funct Integr Genomics 16, 383–398 (2016). https://doi.org/10.1007/s10142-016-0491-2

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