Rice Genomics and Genetics 2025, Vol.16, No.2, 86-95 http://cropscipublisher.com/index.php/rgg 91 6.2 Application in hybrid breeding If breeders consider simplifying the process and improving the efficiency of seed production, then OsTms6 is a very good choice. The "two-line method" breeding using the TGMS line does not require a maintenance line, which is easier to operate (Lee et al., 2005). At the same time, the genetic information and positioning research of OsTms6 itself also give people a clearer understanding of its mechanism of action, which provides a basis for the development of new TGMS lines (Lee et al., 2005). Moreover, similar genes, such as TMS5, can be precisely edited by CRISPR/Cas9, which also means that similar methods are likely to work on OsTms6 and can speed up the generation of "non-transgenic" breeding materials (Zhou et al., 2016). 6.3 Field performance and trait stability However, whether a line can be truly used depends on its performance in the field. TGMS materials with OsTms6 have shown strong drought resistance in actual environments, which is an advantage for areas with tight water resources (Wen et al., 2021). Some studies have also found that the transcription factor OsAL5, which regulates similar genes (such as OsTMS5), can also enhance the drought resistance and temperature sensitivity of plants if it is highly expressed. Although this conclusion is based on other genes, the idea is the same, indicating that OsTms6 may also be able to enhance its practicality through similar means (Wen et al., 2021). In addition, under suitable environmental conditions, this type of TGMS line can also maintain relatively good fertility, which is also critical for the stable production of hybrid seeds (Kadirimangalam et al., 2019). 6.4 Attempts and prospects of gene editing Regarding OsTms6, the use of CRISPR/Cas9 tools has given everyone a lot of room for imagination. Previously, someone had successfully performed site-directed mutagenesis on TMS5 using this technology, indicating that it is not a fantasy to use it to manipulate OsTms6 (Figure 2) (Zhou et al., 2016). As long as the target site is designed in the coding region of this gene, its function can be accurately changed, and even new traits can be introduced. The TGMS strains produced in this way are not only fast to develop, but also avoid transgenic problems and meet the requirements of commercial promotion (Zhou et al., 2016). With the continuous deepening of the understanding of the relevant mechanisms of TGMS, the application value of OsTms6 may be further explored (Jiang et al., 2015; Fan and Zhang, 2017). Figure 2 Pollen fertility of TGMS lines induced by the TMS5ab construct at different temperatures (Adopted from Zhou et al., 2016)
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