RNA-Sequencing Analysis Reveals Critical Roles of Hormone Metabolism and Signaling Transduction in Seed Germination of Andrographis paniculata
Andrographis paniculata is a traditional medicinal plant widely used in South-east Asia. It is cultivated from seed; however, seed germination—even of viable seeds—is unreliable. Understanding the factors controlling germination would enable growers to devise means to improve the efficiency of cultivation. In this study, the transcriptome changes at three time points in A. paniculata seed germination process, namely 0, 28, and 48 h after sowing were analyzed using high-throughput mRNA-sequencing assay (RNA-seq). A total of 198.5 million clean reads were generated and assembled into 84,749 unigenes by merging the de novo and reference assemblies. Transcriptome comparison analysis revealed that the majority of transcriptome changes occurred in the early stage of the germination process. Gene ontology (GO) and Kyoto Encyclopedia of Gene and Genomes (KEGG) pathway analyses indicated that the differentially expressed genes are mainly involved in butanoate, galactose, glycerophospholipid, and carbon metabolism, and in plant hormone signal transduction. Remarkably, the expression levels of genes involved in gibberellin, abscisic acid and ethylene metabolism, and signal transduction were altered in the germination process. Expression of genes related to gibberellin catabolism was down-regulated, expression of genes related to ethylene signal transduction was activated. Especially, in the initial stage of germination, there was a high expression level of NCED5. This abscisic acid biosynthesis-related gene may play a key role in restraining seed germination. The transcriptional expression levels of related genes were verified by qRT-PCR analysis. That means the critical roles of hormone metabolism and signaling transduction in the germination of A. paniculata seeds were found. On a practical level, the results suggest that gibberellin or/and ethylene treatment in combination with presoaking seeds in warm water may effectively promote germination. This study is the first global overview of gene expression involved in seed germination of A. paniculate and provides transcriptome data for further research on the molecular regulation mechanism of seed germination.
KeywordsTranscriptome analysis Seed germination Andrographis paniculata
This work was supported by the National Natural Science Foundation of China (Grant No. 31401277); the Youth Elite Project of Guangzhou University of Chinese Medicine (Grant No. QNYC20140113).
Compliance with Ethical Standards
Conflict of interest
All authors declare no conflict of interest.
- Arc E, Sechet J, Corbineau F, Rajjou L, Marion-Poll A (2013) ABA crosstalk with ethylene and nitric oxide in seed dormancy and germination. Front Plant Sci 4:63Google Scholar
- Chinese Pharmacopoeia Commission (2015) Pharmacopoeia of the People’s Republic of China. Chinese Pharmacopoeia Commission, Beijing p 268Google Scholar
- Haas BJ, Papanicolaou A, Yassour M, Grabherr M, Blood PD, Bowden J, Couger MB, Eccles D, Li B, Lieber M, MacManes MD, Ott M, Orvis J, Pochet N, Strozzi F, Weeks N, Westerman R, William T, Dewey CN, Henschel R, LeDuc RD, Friedman N, Regev A (2013) De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis. Nat Protoc 8:1494–1512CrossRefGoogle Scholar
- He R, Tong JY, Zhang XL, Zhan RT, Chen WW (2011) Study on the quality grading standard of Andrographis paniculata seeds. Med Plant 8:17–20Google Scholar
- Kumari A, Lal RK, Singh KLB (2012) Comparative study of seed germination and seed vigour test in Andrographis paniculata (Acanthaceae). Bot Serbica 36:49–52Google Scholar
- Li LM, Li M (2011) Study on the effects of different treatments on the germination of Andrographis paniculata seed. J Guangdong Pharm Univ 27:371–374Google Scholar
- LI ZJ, GUO JX, LI YW, HE R, WEI JS, XU H (2013) Selection of reference genes for real-time quantitative PCR in Andrographis paniculata (Burm. f.) Nees. J Guangzhou Univ Tradit Chin Med 2:240–244Google Scholar
- Linkies A, Muller K, Morris K, Tureckova V, Wenk M, Cadman CS, Corbineau F, Strnad M, Lynn JR, Finch-Savage WE, Leubner-Metzger G (2009) Ethylene interacts with abscisic acid to regulate endosperm rupture during germination: a comparative approach using Lepidium sativum and Arabidopsis thaliana. Plant Cell 21:3803–3822CrossRefGoogle Scholar
- Rawat R, Vashistha DP (2011) Seed germination improvement in Andrographis paniculata. Int J Med Aromat Plants 3:348–350Google Scholar
- Talei D, Kadir MA, Yusop MK, Valdiani A, Abdullah MP (2012) Physico-protein based dormancy in medicinal plant of Andrographis paniculata. J Med Plants Res 6:2170–2177Google Scholar
- Tong JY, Zhang XL, He R, Zhan RT (2011) Study on germination testing standardization of Andrographis paniculata (Burm. f.) Nees seed. Seed 30:1–3Google Scholar
- Valdiani A, Talei D, Lattoo SK, Ortiz R, Rasmussen SK, Batley J, Rafii MY, Maziah M, Sabu KK, Abiri R, Sakuanrungsirikul S, Tan SG (2017) Genoproteomics-assisted improvement of Andrographis paniculata: toward a promising molecular and conventional breeding platform for autogamous plants affecting the pharmaceutical industry. Crit Rev Biotechnol 37:803–816CrossRefGoogle Scholar
- Zhang XR (2002) WHO monographs on selected medicinal plants. World Health Organization, Geneva, p 12Google Scholar