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Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (10): 2045-2052.doi: 10.3724/SP.J.1006.2021.02079

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Cloning and functional identification of gene OsATS in rice

LI Xiao-Xu(), WANG Rui(), ZHANG Li-Xia, SONG Ya-Meng, TIAN Xiao-Nan, GE Rong-Chao*()   

  1. College of Life Science, Hebei Normal University, Shijiazhuang 050024, Hebei, China
  • Received:2020-11-21 Accepted:2021-03-22 Online:2021-10-12 Published:2021-04-07
  • Contact: GE Rong-Chao E-mail:jiandanxiaoxiao@163.com;1915435558@qq.com;grcgp@sina.com
  • Supported by:
    National Natural Science Foundation of China(30900104);Natural Science Foundation of Hebei Province(C2016205158)

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Abstract:

The plant embryo specific protein ATS3 is closely related to osmotic stress response in plants. Here, the stress resistance related gene OsATS was preliminarily studied in rice. Fluorescence quantitative PCR showed that the relative expression level of OsATS increased significantly after salt stress in rice. The overexpression vector of OsATS was constructed and transformed into Arabidopsis thaliana. The stress resistance test revealed that the overexpression of OsATS gene could significantly improve the salt tolerance of Arabidopsis thaliana at germination and adult stages. After that, the overexpression vector p1300-35s:OSATS and RNA interference vector pTCK303-OsATS-RNAi were transferred into rice. The stress tolerance analysis indicated that the salt tolerance of OsATS overexpression rice lines significantly increased at germination stage and seedling stage, while the salt tolerance of OsATS RNAi rice lines significantly decreased. Results of qRT-PCR and physiological index detection demonstrated that the relative expression levels of OSATS gene might regulate the protein content of proline and LEA cells by regulating the expression of OsP5CS1, OsLEA3-1 and OsPDH, thus affecting the salt tolerance in rice. This study preliminarily revealed the stress resistance function of OSATS gene, which laid a foundation for improving rice stress resistance by adjusting the relative expression level of OSATS gene.

Key words: rice, OsATS genes, overexpression, RNA interference, physiological indexes

Fig. 1

Relative expression levels of OsATS genes under NaCl stress in rice The significant difference is evaluated by the Student’s t-test. *P< 0.05, **P < 0.01."

Fig. 2

Subcellular localization of OsATS in tobacco epidermis"

Fig. 3

PCR detection of OsATS overexpression in Arabidopsis"

Fig. 4

Relative expression levels of OsATS genes in Arabidopsis The significant difference is evaluated by the Student’s t-test. *** P < 0.001."

Fig. 5

Seed germination rate of OsATS overexpression under the salt stress in Arabidopsis The significant difference is evaluated by the Student's t-test. * P < 0.05, ** P < 0.01."

Fig. 6

Salt tolerance detection of OsATS overexpression of adult plants in Arabidopsis"

Fig. 7

Relative expression levels of OsATS overexpressed and OsATS-RNAi transgenic plants in rice WT: wild type rice; OX: OsATS overexpression transgenic rice lines; Ri: RNAi transgenic rice lines; The significant difference is evaluated by the Student’s t-test. *** P < 0.001."

Fig. 8

Seed germination rate of OsATS overexpressed and OsATS-RNAi transgenic plants under NaCl stress in rice"

Fig. 9

Salt tolerance detection of the OsATS transgenic rice"

Fig. 10

Physiological indexes detection of the OsATS overexpression and OsATS-RNAi transgenic plants in rice A: proline content; B: MDA content; C: relative electrolyte leakage; The significant difference is evaluated by the Student’s t-test. * P< 0.05, ** P < 0.01, *** P < 0.001."

Fig. 11

Relative expression levels of the salt tolerance related genes in OsATS transgenic rice The significant difference is evaluated by the Student’s t-test. * P < 0.05, ** P < 0.01, *** P < 0.001."

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