Inhibitory effect of StCYP707A1 gene on tuberization in transgenic potato

다국어 초록 (Multilingual Abstract)

Tuber formation of potato is a complex biological process and is regulated by many factors including phytochrome, hormones, transcription factors, RNAs, microRNAs, etc. In our previous study, CYP707A1, an ABA 80-hydroxylase gene, was down-regulated in the StCOL antisense transgenic potato stolons, but its relation with potato tuberization has not been studied yet. In this study, to investigate the role of this gene in potato tuberization, we cloned a CYP707A1 gene from potato cultivar De ´sire ´e and constructed the StCYP707A1 over-expression and anti-sense potato lines by Agrobacterium-mediated transformation. In total, four over-expression transgenic lines and nine anti-sense transgenic lines were confirmed by PCR analysis. Semi-quantitative RT-PCR and qRTPCR analysis showed that the level of StCYP707A1 transcripts was significantly higher in the over-expression lines (A7 and A9) and lower in the anti-sense lines (F2 and F8), compared to the wild-type control. We further analyzed tuber formation in the transgenic lines and the wild-type control. The result showed that tuber yield per plant and average tuber weight were decreased in A7 and A9 and increased in F2. We also measured the content of ABA and GA3 in transgenic lines. ABA level was reduced in A7 and A9 and increased in F2. Contrariwise, the concentration of GA3 was higher in A7 and A9, and lower in F2 than in wild-type control. These results indicate that StCYP707A1 negatively affects potato tuberization through ABA regulation on gibberellic acid concentration.

참고문헌 (Reference)

1 Xu X, "The role of gibberellin, abscisic acid, and sucrose in the regulation of potato tuber formation in vitro" 117 (117): 575-584, 1998

2 Wang Y, "The role of abscisic acid in regulating cucumber fruit development and ripening and its transcriptional regulation" 64 (64): 70-79, 2013

3 Garcı´a MNM, "The protein phosphatase 2A catalytic subunit StPP2Ac2b acts as a positive regulator of tuberization induction in Solanum tuberosum L" 93 (93): 227-245, 2017

4 Martinez-Garcia JF, "The interaction of gibberellins and photoperiod in the control of potato tuberization" 20 : 377-386, 2002

5 Lin T, "The impact of the long-distance transport of a BEL1-like messenger RNA on development" 161 : 760-772, 2013

6 Kushiro T, "The Arabidopsis cytochrome P450CYP707A encodes ABA 80-hydroxylases: key enzymes in ABA catabolism" 23 (23): 1647-1656, 2004

7 Hofgen R, "Storage of competent cells for Agrobacterium transformation" 16 : 9877-, 1988

8 Ji K, "SlNCED1 and SlCYP707A2: key genes involved in ABA metabolism during tomato fruit ripening" 65 (65): 5243-5255, 2014

9 Hannapel DJ, "Potato biology and biotechnology: advances and perspectives" Elsevier Publishing 237-256, 2007

10 Gonzalez-Schain ND, "Potato CONSTANS is involved in photoperiodic tuberization in a graft-transmissible manner" 70 : 678-690, 2012

11 Hwang Bae Sohn, "Overexpression of jasmonic acid carboxyl methyltransferase increases tuber yield and size in transgenic potato" 한국식물생명공학회 5 (5): 27-34, 2011

12 Jackson SD, "Multiple signaling pathways control tuber induction in potato" 119 : 1-8, 1999

13 Zeevaart JAD, "Metabolism and physiology of abscisic acid" 39 : 439-473, 1988

14 Guivarc’h A, "Local expression of the ipt gene in transgenic tobacco (Nicotiana tabacum L. cv. SR1) axillary buds establishes a role for cytokinins in tuberization and sink formation" 53 (53): 621-629, 2002

15 Garcı´a MNM, "Heterologous expression of Arabidopsis ABF4 gene in potato enhances tuberization through ABA-GA crosstalk regulation" 239 : 615-631, 2014

16 Hu J, "Genetic analysis of sweetpotato and wild relatives using intersimple sequence repeats (ISSRs)" 53 : 297-304, 2003

17 Abelenda JA, "From the model to the crop:genes controlling tuber formation in potato" 22 (22): 287-292, 2011

18 Cutler AJ, "Formation and breakdown of ABA" 4 : 472-478, 1999

19 Chono M, "Field studies on the regulation of abscisic acid content and germinability during grain development of barley: molecular and chemical analysis of pre-harvest sprouting" 57 (57): 2421-2434, 2006

20 Hong Y, "Expression of the MIS-99m gene in transgenic potato plants confers resistance to Phytophthora Infestans and Ralstonia Solanacearum" 31 (31): 418-424, 2013

21 Destefano-Beltra´n L, "Effects of post harvest storage and dormancy status on ABA content, metabolism, and expression of genes involved in ABA biosynthesis and metabolism in potato tuber tissues" 61 (61): 687-697, 2006

22 Sarkar D, "Cytokinins antagonize the jasmonates action on the regulation of potato (Solanum tuberosum)tuber formation in vitro" 87 : 285-295, 2006

23 Abdala G, "Changes in jasmonate and gibberellin levels during development of potatoplants (Solanum tuberosum)" 36 : 121-126, 2002

24 Saito S, "Arabidopsis CYP707As encode (?)-abscisic acid 80-hydroxylase, a key enzyme in the oxidative catabolism of abscisic acid" 134 (134): 1439-1449, 2004

25 Nitsch LMC, "Abscisic acid levels in tomato ovaries are regulated by LeNCED1 and SlCYP707A1" 229 (229): 1335-1346, 2009

26 Mizutani M, "ABA 80-hydroxylase and its chemical inhibitors" 5 (5): 385-404, 2006

27 Murashige T, "A revised medium for rapid growth and bioassays with tobacco tissue cultures" 15 : 473-497, 1962

28 Krochko JE, "(?)-Abscisic acid 80-hydroxylase is a cytochrome P450monooxygenase" 118 : 849-860, 1998