Skip to main navigation menu Skip to main content Skip to site footer
ASPHC Baner

Vol. 17 No. 6 (2018)

Articles

IDENTIFICATION AND EXPRESSION OF GA-RELATED GENES ASSOCIATED WITH in vitro MICRO-TUBERS FORMATION IN Pinellia ternata

DOI: https://doi.org/10.24326/asphc.2018.6.9
Submitted: December 20, 2018
Published: 2018-12-20

Abstract

Pinellia ternata is one of the most important herbs in traditional Chinese medicine. Recently, its yield is hard to meet the market demand, therefore the system of in vitro micro-tubers was developed for its propagation and endogenous GA was revealed probably function negatively during micro-tubers induction. However, the mechanism is still unknown. In this study, 70 mg/L chlormequat chloride (CCC) was successfully used to inhibit the endogenous GA content and promote the micro-tubers induction. Subsequently, suppression subtractive hybridization (SSH) was performed to identify GA-related genes associated with in vitro micro-tubers formation in P. ternata. The cDNAs of micro-tubers induced with and without CCC were used as the “tester”, and “driver”, respectively. SSH library sequencing yielded 300 expressed sequence tags (ESTs). Finally, 226 ESTs were retained after screening, 84 of which had no significant homology to any of previously identified genes and 39 of the remaining 142 ESTs represented singletons. Realtime quantitative RT-PCR analysis of the expression patterns showed that all 5 transcripts showed signal alteration during the process of in vitro micro-tubers formation. The sequences appeared to be highly homologous with 60S ribosomal protein, 26S ribosomal RNA gene, zinc transporter protein, 12kD storage protein and malate dehydrogenase, respectively. These results would facilitate the functional characterization of the GA-related genes associated with in vitro micro-tubers development and subsequent in vitro manipulation.

References

  1. Agrawal, L., Chakraborty, S., Jaiswal, D.K., Gupta, S., Datta, A., Chakraborty, N. (2008). Comparative proteomics of tuber induction, development and maturation reveal the complexity of tuberization process in potato (Solanum tuberosum L.). J. Proteome Res., 7, 3803–3817.
  2. Carpenter, M.A., Joyce, N.I., Genet, R.A., Cooper, R.D., Murray, S. R.,Noble, A.D., Butler, R.C., Timmerman-Vaughan, G.M. (2015). Starch phosphorylation in potato tubers is influenced by allelic variation in the genes encoding glucan water dikinase, starch branching enzymes I and II, and starch synthase III. Front. Plant Sci., 6, 143.
  3. Chang, L., Xu, Y.M. Xue, J.P. (2007). Formation of microtubers from the petiole of Pinellia ternata (Thunb.) Berit. in vitro and change of endogenous hormones content. J. Huazhong Agric. Univ., 05.
  4. Cui, X., Li, X., Meng, F. (2009). Progress on research of rapid propagation of Pinellia ternata. Chinese Agric. Sci. Bull., 25(1), 51–55.
  5. Dammann, R., Lucchini,R., Koller, T., Sogo, J.M. (1993). Chromatin structure and transcription of rDNA in yeast Saccharomyces cerevisiae. Nucleic Acids Res., 21, 2331–2338.
  6. Duan, Y., Xue, T., Li, J., Teng, J., Zhang, A., Sheng, W., Zhu, Y., Chang, L., Xue J., (2015). In vitro induction of yacon tuberous root and identification of genes associated with tuberous root expansion. Japs J. Anim. Plant Sci., 25(6), 1753−1763.
  7. Elrys, A.S., Abdo, A.I.E., Desoky, E.M. (2017). Potato tubers contamination with nitrate under the influence of nitrogen fertilizers and spray with molybdenum and salicylic acid. Environ Sci. Pollut Res. Int., 25(7), 7076−7089.
  8. Gombodorj, S., Yang, M.H., Shang, Z.C., Liu, R. H., Li, T. X., Yin, G.P., Kong, L.Y. (2017). New phenalenone derivatives from Pinellia ternata tubers derived Aspergillus sp. Fitoterapia, 120, 72–78.
  9. Han, M.H., Yang, X.W., Zhang, M., Zhong, G.Y. (2006). Phytochemical study of the rhizome of Pinellia ternata and quantification of phenylpropanoids in commercial Pinellia Tuber by RP-LC. Chromatographia, 64, 647–653.
  10. http://www.ncbi.nlm.nih.gov/BLAST
  11. Li, J., Fang, L., Xue, J. Sheng, W. (2010). [Effects of Pinellia ternata extracts on inhibiting of oviposition and ovicidal action against Plutella xylostella]. Zhongguo Zhong Yao Za Zhi, 35, 1108 –1111 [in Chinese].
  12. Lu, H., Xue, T., Zhang, A., Sheng, W., Zhu, Y. et al., (2013). Construction of an SSH Library of Pinellia ternata under heat stress, and expression analysis of four transcripts. Plant Mol. Biol. Rep., 31, 185–194.
  13. Lucchini, R., Sogo, J.M. (1992). Different chromatin structures along the spacers flanking active and inactive Xenopus rRNA genes. Mol. Cell. Biol., 12, 4288.
  14. Luo, L., Wang, J.N., Kong, L.D., Jiang, Q.G., Tan, R.X. (2000). Antidepressant effects of Banxia Houpu decoction, a traditional Chinese medicinal empirical formula. J. Ethnopharmacol., 73, 277–281.
  15. Qi, P., (2012). Research progress of ZIP transporters gene family. Biotechnol. Bull., 10, 15–18.
  16. Shan, J., Song, W., Zhou, J., Wang, X., Xie, C. et al. (2013). Transcriptome analysis reveals novel genes potentially involved in photoperiodic tuberization in potato. Genomics, 102, 388–396.
  17. Sheng, W., J. Xue, A. Zhang, L. Chang and C. Fan, (2010). [Variation of endogenous hormones in formation of microtubers of Pinellia ternata in vitro]. China J. Chinese Mater. Med., 51, 4 –5 [in Chinese].
  18. Shi, K., Wu, X., Ma, J., Zhang, J., Zhou, L. et al. (2017). Effects of planting and processing modes on the degradation of dithianon and pyraclostrobin in Chinese yam (Dioscorea spp.). J. Agric. Food Chem. 65, 10439–10444.
  19. Sun, P., Guo, Y., Qi, J., Zhou, L., Li, X. (2010). Isolation and expression analysis of tuberous root development related genes in Rehmannia glutinosa. Mol. Biol. Rep., 37, 1069–1079.
  20. Tsai, Y.T., Lin, C.I., Chen, H.K., Lee, K.M., Hsu. C.Y. et al. (2008). Chromatin tethering effects of hNopp140 are involved in the spatial organization of nucleolus and the rRNA gene transcription. J. Biomed. Sci., 15, 471–486.
  21. Wang, X., (2006). Research progress of plant malate dehydrogenase. Biotechnol. Bull., 4, 44–47.
  22. Wu, X., Xiong, E., An, S., Gong, F., Wang, W. (2012). Sequential extraction results in improved proteome profiling of medicinal plant Pinellia ternata tubers, which contain large amounts of high-abundance proteins. PLoS One, 7, e50497.
  23. Xiao-Yan, L.I., Dong-Ming, Q.I., Sun, C.R. (2008). Anti-feeding activitiy of ethanol extracts from Pinellia Ternata Breit against Henosepilachna vigintioctopunctata (Fabricius). J. China West Normal Univ., 29(1), 59–62.
  24. Xu, J.Y., Dai, C., Shan, J.J., Xie, T., Xie, H.H. et al. (2018). Determination of the effect of Pinellia ternata (Thunb.) Breit. on nervous system development by proteomics. J. Ethnopharmacol., 213, 221–229.
  25. Xue, J., Zhu, Y., Aimin, Z. Liu, J. (2004). Research on direct formation of microtubers from Pinellia ternata. Acta Agron. Sin., 30, 1060–1064.
  26. Xue, T., Dai, X., Wang, R., Wang, J., Liu, Z. et al. (2017). ARGONAUTE10 inhibits in vitro shoot regeneration via repression of miR165/166 in Arabidopsis thaliana. Plant Cell Physiol., 58, 1789.
  27. You, M.K., Hur, C.G., Ahn, Y.S., Suh, M.C., Jeong, B.C. et al. (2003). Identification of genes possibly related to storage root induction in sweet potato. Febs Lett., 536, 101.
  28. Zhang, S., Xie, S. (1998). Study on tissue culture and rapid propagation of Pinellia ternata. J. Xinyang Teach. Coll., 11(1), 86–88.
  29. Zhang, Y.N., He, P., Xue, J.P., Guo, Q., Zhu, X.Y. et al. (2017). Insecticidal activities and biochemical properties of Pinellia ternata extracts against the beet armyworm Spodoptera exigua. J. Asia-Pacific Entomol., 20, 469–476.

Downloads

Download data is not yet available.

Most read articles by the same author(s)

Similar Articles

<< < 1 2 3 4 5 6 7 8 9 10 > >> 

You may also start an advanced similarity search for this article.