Research articleOsGERLP: A novel aluminum tolerance rice gene isolated from a local cultivar in Indonesia
Graphical abstract
Introduction
Rice (Oryza sativa L.) is an essential staple food in Indonesia and one of the largest food crops in the world. Its intensive cultivation is a common agricultural practice that aims at high yield. However, the decreasing paddy fields due to rapid conversion into settlement and industrial uses significantly limit rice production. An alternative solution to this problem is the extension of rice cultivation area to suboptimal lands, such as acid soil, which is abundant in Indonesia. However, high aluminum (Al) solubility in soil with a pH of less than 5 negatively affects rice cultivation in acid soil (Delhaize and Ryan, 1995; Delhaize et al., 2001). Al ions in the rhizosphere damage the root system (Ma and Hiradate, 2000), making plants more vulnerable to drought and mineral nutrition deficiency. These conditions inhibit growth and decrease production (Matsumoto, 2000).
The use of Al-tolerant varieties adapted to acid soil would significantly solve the Al toxicity problem worldwide. In Indonesia, several local cultivars are tolerant to Al, such as Grogol, Krowal, and Hawara Bunar, though they are not high yielding (Miftahudin et al., 2007; Miftahudin and Chikmawati, 2008). Hawara Bunar is considered the most Al-tolerant rice cultivar. Hence, it can be used to develop high yielding rice varieties tolerant to Al and acid soil.
Several genes control the Al tolerance trait in rice (Nguyen et al., 2003), and some of them have been identified and isolated (Yamaji et al., 2009; Tsutsui et al., 2011; Yokosho et al., 2011; Li et al., 2014). Miftahudin et al. (2005) reported microsynteny between an Al tolerance locus on rye (Secale cereale L.) chromosome arm 4RL and a BAC clone OSJNBa0091P11 from rice chromosome 3. Two markers, BCD1230 and B6, border both regions. Additionally, we identified a potential Al-tolerance candidate gene between markers in the rice BAC clone.
Citrate secretion contributes to the Al tolerance mechanisms in rice (Ma et al., 2005). The OsFRDL4 gene, which is upregulated by Al, controls the citrate secretion from the rice roots (Yokosho et al., 2011). According to Ma et al. (2014), the transcription factor, ART1, controls the OsFRDL4 gene expression. However, further analysis showed that Al did not regulate the ART1 gene. This indicates that an unknown upstream gene may regulate the ART1 gene. To understand Al tolerance mechanisms in rice, it is necessary to clone the new Al tolerance candidate gene from rice chromosome and analyze its role in controlling the Al-tolerance trait and regulating Al-related gene expression.
This paper reports on the cloning and characterization of an Al tolerance candidate gene, OsGERLP (GenBank: MH388027.1). The gene is isolated from rice cv. Hawara Bunar, a local Indonesian rice cultivar, using the microsynteny relationship between rye and rice. The gene's responses of the OsGERLP-silenced transgenic rice to Al stress are evaluated. Furthermore, the role in enhancing Al tolerance in transgenic tobacco (Nicotiana tabacum L.) is also evaluated.
Section snippets
Plant materials
This experiment used Rice cv. Hawara Bunar, IR64, and the F9 Recombinant Inbred Line Population derived between IR64 and Hawara Bunar, as well as T3 transgenic tobacco and its wild-type seeds. The T3 transgenic tobacco seeds with a homozygous OsGERLP gene were obtained from T2 plants. T1 transgenic rice seeds containing 3-UTR of the OsGERLP gene were used to generate OsGERLP-silenced T2 transgenic plants (Wahyuningtyas et al., 2016). The OsGERLP-silenced T2 homozygous plants were then used for
Rice root length characters under aluminum stress
The Al-tolerant rice cv. Hawara Bunar shows a higher TRL than the Al-sensitive rice cv. IR64 (Fig. 1). In previous studies, the TRL and PRL are Al tolerance characters which have be used to distinguish between several Al-tolerant and sensitive local cultivars of Indonesian rice (Siska et al., 2017; Fendiyanto et al., 2019a). To reconfirm the Al tolerance level of rice cv. Hawara Bunar and IR64, they were evaluated using PRL and TRL characters under 555 μM Al stress. Al-treated rice cv. Hawara
Discussion
Rice shows different tolerance levels to Al toxicity among cultivars. Data showed that rice cv. Hawara Bunar is more tolerant to Al than cv. IR64, supported by root length characters, Al accumulation, and citrate secretion data (Fig. 1). Additionally, based on the primary root length (PRL) and total root length (TRL) parameters, rice cv. Hawara Bunar shows only a slight decrease in both root length characteristics between Al-treated and control conditions. Conversely, rice cv. IR64 showed a
Author contributions
M.M. is the PI of the research project, designed and conducted the experiment, drafted, edited, and finalized the manuscript. T.C.H., S.S designed, conducted the experiment, and edited the manuscript. H.T.N. and J.P.G. collaborators, partially designed the experiment, provided facilities and expression vector, and edited the manuscript. D.I.R., A.H. identified, isolated, partially characterized the OsGERLP in rice, and transformed the tobacco. M.H.F. and R.D.S. conducted QTL mapping and
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgment
The research was funded by the International Collaboration and Scientific Publication Research Grant, F.Y. 2014–2016, and World Class Research (WCR) Grant, F.Y. 2019 awarded to Dr. Miftahudin from the Indonesian Ministry of Research and Technology, the Republic of Indonesia. We thank Dr. Utut Widyastuti for her suggestions and technical assistance in the molecular biology laboratory work.
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