Abstract
Introduction Salinity is a complex environmental stress that affects the growth and production of rice worldwide. But there are some rice landraces in coastal regions that can survive in presence of highly saline conditions. An understanding of the molecular attributes contributing to the salinity tolerance of these genotypes is important for developing salt-tolerant high yielding modern genotypes to ensure food security. Therefore, we investigated the role and functional differences of two K + salt-responsive transporters. These are OsTPKa or Vacuolar two-pore potassium channel and OsHAK_like or a hypothetical protein of the HAK family. These transporters were selected from previously identified QTLs from the tolerant rice landrace genotype (Horkuch) and sensitive genotype (IR29). Methods In silico comparative sequence analysis of the promoter sequences of two these genes between Horkuch and IR29 was done. Real-Time expression of the selected genes in leaves and roots of IR29 (salt-sensitive), I-14 and I-71 (Recombinant Inbred Lines of IR29(♀)× Horkuch), Horkuch and Pokkali (salt-tolerant) under salt-stress at different time points was analyzed. For further insight, OsTPKa and OsHAK_like were chosen for loss-of-function genomic analysis in Horkuch using the CRISPR/Cas9 tool. Furthermore, OsTPKa was chosen for cloning into a sensitive variety by Gateway technology to observe the effect of gain-of-function. Results The promoter sequences of the OsTPKa and OsHAK_like genes showed some significant differences in promoter sequences which may give a survival advantage to Horkuch under salt-stress. These two genes were also found to be overexpressed in tolerant varieties (Horkuch and Pokkali). Moreover, a coordinated expression pattern between these two genes was observed in tolerant Horkuch under salt-stress. Independently transformed plants where the expression of these genes was significantly lowered, performed poorly in physiological tests for salinity tolerance. On the other hand, positively transformed T 0 plants with the OsTPKa gene from Horkuch consistently showed growth advantage under both control and salt stress. Discussion The poor performance of the transgenic plants with the down-regulated genes OsTPKa and OsHAK_like under salt stress supports the assumption that OsTPKa and OsHAK_like play important roles in defending the rice landrace Horkuch against salt stress, minimizing salt injury, and maintaining plant growth. Moreover, the growth advantage provided by overexpression of the vacuolar OsTPKa K + transporter, particularly under salt stress reconfirms its important role in providing salt tolerance. The QTL locus from Horkuch containing these two transporters maybe bred into commercial rice to produce high-yielding salt tolerant rice.
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Claims 7
Claim · model-written summary
OsTPKa promoter sequences differed between Horkuch and IR29.
- Taxon
- rice
Oryza sativat0 - Treatment
- salt-stresse0
- Gene
- OsTPKae0
- Outcome
- differences in promoter sequencese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The promoter sequences of the OsTPKa and OsHAK_like genes showed some significant differences in promoter sequences which may give a survival advantage to Horkuch under salt-stress.”
t0“rice”
Claim · model-written summary
OsHAK_like promoter sequences differed between Horkuch and IR29.
- Taxon
- riceOryza sativat0
- Treatment
- salt-stresse0
- Gene
- OsHAK_likee0
- Outcome
- differences in promoter sequencese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The promoter sequences of the OsTPKa and OsHAK_like genes showed some significant differences in promoter sequences which may give a survival advantage to Horkuch under salt-stress.”
t0“rice”
Claim · model-written summary
OsTPKa was overexpressed in tolerant varieties.
- Taxon
- riceOryza sativat0
- Material
- Horkuch and Pokkalie0
- Gene
- OsTPKat1
- Outcome
- overexpressede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“These two genes were also found to be overexpressed in tolerant varieties (Horkuch and Pokkali).”
t0“rice”
t1“These are OsTPKa or Vacuolar two-pore potassium channel and OsHAK_like or a hypothetical protein of the HAK family.”
Claim · model-written summary
OsHAK_like was overexpressed in tolerant varieties.
- Taxon
- riceOryza sativat0
- Material
- Horkuch and Pokkalie0
- Gene
- OsHAK_liket1
- Outcome
- overexpressede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“These two genes were also found to be overexpressed in tolerant varieties (Horkuch and Pokkali).”
t0“rice”
t1“These are OsTPKa or Vacuolar two-pore potassium channel and OsHAK_like or a hypothetical protein of the HAK family.”
Claim · model-written summary
The two genes showed coordinated expression in Horkuch under salt stress.
- Taxon
- riceOryza sativat0
- Material
- Horkuche0
- Treatment
- salt-stresse0
- Outcome
- coordinated expression patterne0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a coordinated expression pattern between these two genes was observed in tolerant Horkuch under salt-stress.”
t0“rice”
Claim · model-written summary
Lowered expression of the two genes accompanied poor performance in salinity tolerance tests.
- Taxon
- riceOryza sativat0
- Treatment
- expression of these genes was significantly lowerede0
- Outcome
- physiological tests for salinity tolerancee0
- Direction
- poorlye0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Independently transformed plants where the expression of these genes was significantly lowered, performed poorly in physiological tests for salinity tolerance.”
t0“rice”
Claim · model-written summary
Plants transformed with Horkuch OsTPKa showed growth advantage under control and salt stress.
- Taxon
- riceOryza sativat0
- Treatment
- positively transformed T 0 plants with the OsTPKa gene from Horkuche0
- Gene
- OsTPKae0
- Outcome
- growth advantagee0
- Direction
- advantagee0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“positively transformed T 0 plants with the OsTPKa gene from Horkuch consistently showed growth advantage under both control and salt stress.”
t0“rice”
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