REGULATORY ELEMENTS FOR TRANSAGENTS EXPRESSION IN PLANTS MERISTEMS

Application of promoter sites that provide tissue-specific expression of transgenes, allows accumulating foreign proteins in target tissues. This reduces the risk of bad influence on plants, human and environment. Otherwise, high-specific promoter sites often loose or change their specificity when being transferred to new genetic environment that requires beforehand checking of their expressive effect in transgenic plants by means of reporter genes. The paper aims at evaluating the nature and efficiency of reporter gene uidA expression when being managed by promoter sites of Arabidopsis thaliana, apеtala3 and rpt2a genes and transferring to genome Nicotiana tabacum. Gene apеtala3 is expressed in flower meristems and codes transcription factor of В group, which regulates flower development. Gene rpt2a codes 19S regulatory subunit 26S of proteasome and it is expressed in apical meristems of root and stalk mainly.The article shows that promoter sites of the genes provide tissue-specific expression of reporter gene uidA in conditions of allogeneic genome. The authors observed ectopical expression of transgene in relation to the type of promoter site used. This expression is restricted by meristem areas in the organs of transgenic plants. Comparative analysis of testing results on kanamycin resistance and hystochemical analysis has shown that GUS-coloring intensity was higher in derivatives of tabacco transformants, which carry two and more insertions of transgene in the genome. This speaks about possible additive effect of transgene insertions number that enhances expression and accumulation of reporter gene product in the tissues of transgene tabacco plants.

transgenesis, promoter site, genetic construct, tissue-specific expression, reporter gene, трансгенез, промотор, генетическая конструкция, тканеспецифическая экспрессия, репортерный ген

1. Potenza C., Aleman L., Sengupta-Gopalan Ch. Targeting transgene expression in research, agricultural, and environmental applications: promoters used in plant transformation // In Vitro Cell. Dev. Biol. Plant. - 2004. - Vol. 40. - P. 1-22.

2. Two tobacco AP1-like gene promoters drive highly specific, tightly regulated and unique expression patterns during floral transition, initiation and development / J. Zhang, G. Yan, Z. Wen [et al.] // Planta. - 2014. - Vol. 239. - P. 469-478.

3. Discrete spatial and temporal cis-acting elements regulate transcription of the Arabidopsis floral homeotic gene APETALA3 / Th.A. Hill, Ch.D. Day, S. C. Zondlo [et al.] // Development. - 1998. - Vol. 125. - P. 1711-1721.

4. Krizek B. A., Meyerowitz E. M. The Arabidopsis homeotic genes APETALA3 and PISTILLATA are sufficient to provide the B class organ identity function // Development. - 1996. - Vol. 122. - P. 11-22.

5. Regulation of leaf organ size by the Arabidopsis RPT2a 19S proteasome subunit / Y. Sonoda, K. Sako, Y. Maki [et al.] // The Plant Journal. - 2009. - Vol. 60. - P. 68-78.

6. The HALTED ROOT gene encoding the 26S proteasome subunit RPT2a is essential for the maintenance of Arabidopsis meristems / M. Ueda, K. Matsui, S. Ishiguro [et al.] // Development. - 2004. - Vol. 131. - P. 2101-2111.

7. A simple and general method for transferring genes into plants / R. B. Horsch, J. E. Fry, N. I. Hoffmann [et al.] // Science. - 1985. - Vol. 227. - P. 1229-1231.

8. Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method / X. Zhang, R. Henriques, S. Lin [et al.] // Nature Protocols. - 2006. - Vol. 1. - P. 641-646.

9. Дейнеко Е. В., Загорская А. А., Шумный В. К. Т-ДНК-индуцированные мутации у трансгенных растений // Генетика. - 2007. - Т. 43, № 1. - С. 5-17.

10. Дорохов Ю. Л. Умолкание генов у растений // Молекулярная биология. - 2007. - Т. 41, № 4. - С. 579-592.

11. Маренкова Т. В., Дейнеко Е. В. Инактивирование генов у растений на уровне транскрипции // Генетика. - 2010. - Т. 46, № 5. - С. 581-592.

12. Филипенко Е. А., Дейнеко Е. В., Шумный В. К. Особенности районов встраивания Т-ДНК у трансгенных растений // Генетика. - 2009. - Т. 45, № 11. - С. 1461-1475.

13. Causiera B., Schwarz-Sommerb Z., Daviesa B. Floral organ identity: 20 years of ABCs // Seminars in Cell & Developmental Biology. - 2010. - Vol. 21. - P. 73-79.

14. An intergenic region shared by At4g35985 and At4g35987 in Arabidopsis thaliana is a tissue specific and stress inducible bidirectional promoter analyzed in transgenic arabidopsis and tobacco plant / J. Banerjee, D. K. Sahoo, N. Dey [et al.] [Electronic resource] // PLoS One. - 2013. - Vol. 8, No 11. - e79622. - URL: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3834115.

15. Novel green tissue-specific synthetic promoters and cis-regulatory elements in rice / R. Wang, M. Zhu, R. Ye [et al.] [Electronic resource] // Scientific Reports. - 2015. - Vol. 5:18256. - DOI: 10.1038/srep18256. - URL: https://www.nature.com/articles/srep18256.pdf.