LITERATURE REVIEW ON SALT RESISTANT TRANSGENIC RICE VARIETY IN BANGLADESH

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Literature Review on Salt Resistant Transgenic rice variety in Bangladesh

BRAC UniversityDepartment of Mathematics and Natural Sciences

Submitted by:

Md. Waliullah [email protected]

Salt Resistant Transgenic rice variety in Bangladesh A Literature review

Introduction

Rice is the seed of the grass species Oryza sativa (Asian rice) or Oryza glaberrima (African rice). As a cereal grain, it is the most widely consumed staple food for a large part of the world's human population, especially in Asia including Bangladesh. It is the agricultural commodity with the third-highest worldwide production, after sugarcane and maize, according to 2012 FAOSTAT data.

Since a large portion of maize crops are grown for purposes other than human consumption, rice is the most important grain with regard to human nutrition and caloric intake, providing more than one-fifth of the calories consumed worldwide by humans.

Transgenic Rice or Genetically modified rice are rice strains that have been genetically modified. Rice plants have been modified to increase micronutrients such as vitamin A, accelerate photosynthesis, tolerate herbicides, resist pests, increase grain size, generate nutrients, flavours or produce human proteins.

The natural movement of genes across species, often called horizontal gene transfer or lateral gene transfer, can also occur with rice through gene transfer mediated by natural vectors. However, the cultivation and use of genetically modified varieties of rice remains controversial and is not approved in some countries.

Objective of Transgenic rice production:

Objective of Transgenic rice production is to given bellow-

High-yielding varieties

Flood-tolerant rice

Drought-tolerant rice

Salt-tolerant rice

Environment-friendly rice

Future potential

Golden rice

Expression of human proteins

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LiteratureName

Outcome

01. Zeba I. Seraj et al., (2015); reported Constitutive Overexpression of the Plasma MembraneNa+/H+ Antiporter for Conferring Salinity Tolerance inRice.

The rice variety BRRI dhan28 is popular for high yielding, but salt sensitive, was transformed with the OsSOS1 gene isolated from salt tolerant Pokkali rice and driven by the constitutive promoter, CaMV35S. Future work will contain transformation of both the OsSOS1 and OsNHX1 genes together; with the hope for enhancing the tolerance level compared to currently available transgenic rice. At the cellular level, the Salt Overly Sensitive (SOS) signaling pathway comprising SOS3, SOS2, and SOS1 has been proposed to mediate cellular signaling under salt stress to maintain ion (Na+) homeostasis. The new created transgenic rice was shown to be salt tolerant compared to wild.

02. Zeba I. Seraj et al., (2016); reported Cloning, characterization and analysis of the Arabidopsis RD29A promoter for its inducible expression in rice under salinity and drought stress.

In this study, a stress inducible promoter RD29A from Arabidopsis was characterized by RD29A-GUS construct and transformed in Binnatoa rice variety. Histochemical assay was performed at seedling and reproductive stages of rice to test the inducibility of RD29A promoter under salt stress (100 mM and 200 mM). In both seedling and reproductive stage maximal GUS expression was observed at 24 hours at 100 mM salt stress, whereas GUS expression was found relatively higher under high salt stress condition. This timing concluded that the RD29A promoter is most suitable for driving transgenes for conferring salt tolerance.

03. Zeba I. Seraj et al., (2010) reported Vacuolar Na+/H+ Antiporter Expression and SaltTolerance Conferred by Actin1D and CaMV35S areSimilar in Transgenic Binnatoa Rice

This study shows that the performance of Actin1D and CaMV35S promoter are similar on vacuolar Na+/H+ antiporter (OsNHX1) of rice. Semi‐quantitative RT‐PCR are used to confirms the over‐expression of the gene under both promoters. Salinity tolerance assessment was done at seedling and reproductive stages to compare the performance of the two transgenic lines. The expression of the antiporter gene with the two different promoters resulted in lower K+/Na+ ratios in both the transgenic lines compared to controls, which demonstrates the similar performance of the two promoters to drive the same gene OsNHX1 for reporting salt tolerance in rice.

04. Zeba I. Seraj et al., (2014) reported Cloning and Characterization of

In the current study, with the help of Gateway Technology the upstream regions of the Adh gene from the genomic DNA of Arabidopsis cloned into a promoter characterization binary destination vector and by using PCR,‐ restriction digestion and sequencing a positive clone are confirmed. The construct was then transformed into LBA4404 strain of Agrobacterium by

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A Literature review on Salt resistant transgenic rice variety in Bangladesh


AlcoholDehydrogenase (Adh) Promoter Region for ExpressionUnder Submergence and Salinity Stress

electroporation and then infected to rice calli. In both salt and submergence stresses, Adh could selectively express GUS gene activity compared to control. These results can be further fine-tuned using quantitative measures of GUS activity as well as checking induction in the specific tissues of plants regenerated from the positive calli.

05. Zeba I. Seraj et al., (2009) reported Constitutive expression of OsNHX1 under the promoter Actin1D can improve the salttolerance and yield characteristics of Bangladeshi rice Binnatoa.

The aim of the study is to increasing the vacuolar osmotic potential as it would favor water movement from apoplast to the cell. Production and molecular characterization of OsNHX1-expressing transgenic rice pCAMBIA- 1305.1/Actin1D-OsNHX1construct was produced, cloned in E.coli DH5 , confirmed and then transformed into α Agrobacterium tumefaciens through electroporation. For transgenic status confirmation PCR and Southern blot analysis from T0 to T3 state ware used and over-expression of OsNHX1 was confirmed by semi-quantitative RT-PCR and Western Dot-Blot analyses. After transfection it showed better physiological status and enhanced tolerance and improved yield in salt stress compared to the wild-type.

06. Zeba I. Seraj et al.,(2013) reported Cloning of the Plasma Membrane Sodium/HydrogenAntiporter SOS1 for its Over expression in Rice.

In the current study, the coding sequence of SOS1 gene which is a critical component for salt tolerance among many species was amplified from a rice landrace, Pokkali, by applying the fusion PCR strategy. The amplified product of SOS1 gene was cloned in the pENTR/D TOPO cloning vector‐ and recombined with the binary vector, pH7WG2. The final destination vector was then used to do Agrobacterium mediated transformation. The newly created transgenic rice showed better germination and growth in saline environments due to presence of Na+/H+ antiporter gene.

07. Zeba I. Seraj et al.,(2009) reportedAgrobacterium‐mediated Transformation andConstitutive Expression of PgNHX1 from Pennisetumglaucum L. in Oryza sativa L. cv. Binnatoa

The Pennisetum glaucum vacuolar Na+/H+ antiporter gene (PgNHX1) was transformed by Agrobacterium mediated transformation and expressed in rice Binnatoa under the constitutive promoter CaMV35S. The confirmation of transgenic status of the plant was carried out by PCR and Southern blot hybridization. After successful transformation, transgenic seedlings showed enhanced tolerance at 160 mM NaCl stress compared to controls.

08. Zeba I. Seraj et al.,(2012) reported Over-expression of a DEAD-box helicase, PDH45, confers both seedling and reproductive

A DEAD-box helicase gene ware isolated from pea with a CaMV35S promoter and transformed into Binnatoa rice variety through Agrobacterium mediated transformation. The T2 progenies were considered for salinity tolerance at both seedling and reproductive stages. Under normal physiological conditions transgenic seedlings showed higher chlorophyll content, but

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stage salinity tolerance to rice.

decreased root length compared to wild type. At the reproductive stage, transgenic rice plants showed better fertility and produced higher grain yield then wild type plants under continuous salt stress.

09. Zeba I. Seraj et al.,(2002) reported Agrobacterium-mediated transformation of Bangladesh indica for conferring salt tolerance.

The suitable rice varieties were established by Agrobacterium-mediated transformation techniques with the goal of incorporation of genes for the production of osmoprotective compounds. Mannitol-1-phosphate dehydrogenase for the production of mannitol and trehalose synthase and phosphatase for the production of trehalose was acquired. Transformation experiments using the two above genes are in progress with BR5331 that shows moderately salt tolerant rice variety.

10. Zeba I. Seraj et al.,(2010) reported Enhanced salinity tolerance and improved yield properties in Bangladeshi rice Binnatoa through Agrobacterium-mediated transformation of PgNHX1 from Pennisetum glaucum.

Using Agrobacterium-mediated transformation method transgenic rice cv. Binnatoa was accomplished with the Pennisetum glaucum vacuolar Na+/H+ antiporter gene (PgNHX1) under a constitutive CaMV35S promoter. PCR and RT-PCR were performed for the molecular analysis of putative transgenic plants. The enginerred transgenic rice plants showed better physiological status and completed their life cycle and produced higher grain yields, while wild-type plants exhibited rapid chlorosis and growth inhibition under salt stress.

11. Zeba I. Seraj et al.,(2016) reported Enhanced Salt Tolerance Conferred by the Complete 2.3kb cDNA of the Rice Vacuolar Na+/H+ AntiporterGene Compared to 1.9kb Coding Region with 5 UTR in Transgenic Lines of Rice

In this study, there were two separate transformation events of the OsNHX1 gene, one with 1.9 kb region containing the 5′ UTR with CDS and the other of 2.3 kb, including 5′ UTR, CDS, and the 3′ UTR regions were performed. Tissue culture responsive Binnatoa was transformed with both the constructs separately. Both transgenic lines were observed to be tolerant at both seedling and reproductive stages under salinity stress compared to WT plants. But, the transgenic lines that carry the CDS with both the 5′ and 3′ UTR showed significantly more tolerant compared to the transgenic lines containing OsNHX1gene without the 3′ UTR.

12. Kutubuddin Ali Molla et al., (2015) reported Identification and analysis of novel salt responsive

The present study represents the potential of salt responsive candidate gene based SSR (cgSSR) markers to be utilized as novel and remarkable candidate for diversity analysis among rice genotypes differing in salinity response. The gene bank locus numbers were retrieved and subsequently

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candidate gene based SSRs (cgSSRs) from rice.

sequences of all those genes were downloaded from the web resources of Rice Genome Annotation Project. The primer was designed from the flanking sequences of the identified microsatellite repeat region with some parameters. Allele scoring and sequencing was done by using Alpha View software. After analyzing salt responsive candidate gene based SSR markers ware established and applied to make the salinity tolerance rice.

13. Zeba I. Seraj et al.,(2011) reported Physiology and gene expression of the rice landrace Horkuch under salt stress.

Horkuch is popular salt tolerant rice variety was taken and characterized up to maturity with a modern variety (BRRI dhan41) and (BRRI dhan29) Pokkali under NaCl stress. The expression of salt-responsive genes was investigated by using microarray analysis to understand the physiology at the molecular level. Salt-stressed cDNA of Horkuch seedlings were hybridised with cDNA probes synthesised from Arabidopsis thaliana Heynh. The increased photosynthesis-related genes expression suggested that this could be the reason for the better yield performance of Horkuch.

14. Mohammad G. Mostofa et al., (2015) reported Hydrogen Sulfide Regulates Salt Tolerance in Rice by Maintaining Na+/K+ Balance, Mineral Homeostasis and Oxidative MetabolismUnder Excessive Salt Stress.

The study reveals the importance of the implication of gasotransmitters like H2S for the management of salt stress, thus assisting rice plants to adapt to adverse environmental changes. At the same time, the role of H2S in regulating Na+/K+ balance and mineral homeostasis was also examined. This results support the notion that H2S could serve as an important priming agent in enhancing rice tolerance to salt stress, particularly by inhibiting Na+ uptake, reestablishing redox and mineral homeostases, as well as reducing oxidative stress by stimulating ROS and MG detoxifications.

15. K. Miah et al., (2015); reported Agrobacterium Mediated Genetic Transformation of Rice for Salinity Tolerance for Combating Climate Change.

The objectives of this experiment were to developing a transformation protocol of transgenic rice by using Agrobacterium tumefaciens and transferring the salinity tolerance gene into modern rice cultivars. Salt tolerant transgenic rice was established by injecting mature embryos with Agrobacterium strain LBA4404 that carry a binary vector pCIPK with GUS and nptll gene. This gene was transformed into Binadhan-5 and Binadhan-6 rice varieties. After infection Binadhan-6 showed highest response to GUS assay. The best regeneration of shoots showed when the infection time was 25 minutes and 4 days co-cultivation period. This protocol helps to develop salinity tolerance rice line(s) for combat climate change.

16. Zeba I. Seraj et al.,(2015) reported Salinity and drought tolerance conferred by in planta transformation of SNAC1 transcription

The current study shown that the overexpression of stress-responsive NAC1 transcription factor significantly increases the salinity and drought tolerance transgenic rice. The indica rice variety BRRIdhan 55 was transformed with the SNAC1 TF from the rice landrace Pokkali by using the in planta method. Transformation efficiencies was increased by adding acetosyringone in the Agrobacterium suspension and co-culturing the media. The level of transgene expression was measured by real-time

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factor into a high-yielding rice variety of Bangladesh.

PCR. Phenotypic screening was conducted at T2 and T3 seedling stages where the transgenic lines exhibited much better tolerance compared to their control at 120 mM salt stress.

Conclusions:

Salinity is one of the most critical environmental problems that cause retardation of plant growth by disturbing intracellular ion homeostasis. Soil salinity has been a major threat in crop production worldwide and it exists approximately in 20% of world’s cultivable lands. For a densely populated country like Bangladesh desperately needs substantial increases in crop production to provide her teeming millions with food. The larger portions of land of south western districts of the country are affected by varying degrees of salinity. Agricultural land use of this area is very poor, which is roughly 50% of the country’s average. So, it is necessary to make salt tolerant rice variety to cope with the situation and to increase the productivity of rice. The hope is that, the rice variety BRRI dhan 40, BRRI dhan 41, BRRI dhan 47, BRRI dhan 53, BRRI dhan 54, BRRI dhan 55, BRRI dhan 61 and BRRI dhan 67 has been developed by BRRI that shows better resistance to salt stress as well as high yielding.

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References:

1. Zeba I. Seraj, Farida Yasmin et al., (2015).Constitutive Overexpression of the Plasma Membrane Na+/H+ Antiporter for Conferring Salinity Tolerance in Rice; Plant Tissue Cult. & Biotech. 25(2): 257 272.‐

2. Zeba I. Seraj, Md Muntasir Ali et al., (2016). Cloning, characterization and analysis of the Arabidopsis RD29A promoter for its inducible expression in rice under salinity and drought stress. Bioresearch Communications, 2(1), 139-145.

3. Zeba I. Seraj et al., (2010). Vacuolar Na+/H+ Antiporter Expression and Salt Tolerance Conferred by Actin1D and CaMV35S are Similar in Transgenic Binnatoa Rice; Plant Tissue Culture & Biotech. 19(2): 257-262.

4. Zeba I. Seraj et al., (2014) reported Cloning and Characterization of Alcohol Dehydrogenase (Adh) Promoter Region for Expression Under Submergence and Salinity Stress, Plant Tissue Cult. & Biotech. 24(1): 111-120.

5. Zeba I.Seraj et al., (2009). Constitutive expression of OsNHX1 under the promoter Actin1D can improve the salt tolerance and yield characteristics of Bangladeshi rice Binnatoa. Australian Journal of Crop Science, 3(6):329-335 (2009), ISSN: 1835-2707.

6. Zeba I. Seraj et al.,(2013). Cloning of the Plasma Membrane Sodium/Hydrogen Antiporter SOS1 for its Over expression in Rice. Plant Tissue Culture & Biotech. 23(2): 263 273.‐

07. Zeba I. Seraj et al.,(2009). Agrobacterium mediated Transformation and‐ Constitutive Expression of PgNHX1 from Pennisetum glaucum L. in Oryza sativa L. cv. Binnatoa, Plant Tissue Culture & Biotech. 19(1): 25-33.

08. Zeba I. Seraj et al.,(2012). Over-expression of a DEAD-box helicase, PDH45, confers both seedling and reproductive stage salinity tolerance to rice. Mol Breeding 30:345–354.

09. Zeba I. Seraj et al.,(2002). Agrobacterium-mediated transformation of Bangladesh indica for conferring salt tolerance. Chapter- Prospects for Saline Agriculture; Volume 37, 167-176.

10. Zeba I. Seraj et al.,(2010). Enhanced salinity tolerance and improved yield properties in Bangladeshi rice Binnatoa through Agrobacterium-mediated transformation of PgNHX1 from Pennisetum glaucum.; Acta Physiol Plant (2010) 32:657–663.

11. Zeba I. Seraj et al.,(2016). Enhanced Salt Tolerance Conferred by the Complete 2.3kb cDNA of the Rice Vacuolar Na+/H+ Antiporter Gene Compared to 1.9kb Coding Region with 5 UTR in Transgenic Lines of Rice. Front. Plant Sci. 7:14. doi:10.3389/fpls.2016.00014.

12. Kutubuddin Ali Molla et al., (2015). Identification and analysis of novel salt responsive candidate gene based SSRs (cgSSRs) from rice. DOI 10.1186/s12870-015-0498-1

13. Zeba I. Seraj et al., (2011). Physiology and gene expression of the rice landrace Horkuch under salt stress. Functional Plant Biology; 38(4) 282-292.

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14. Mohammad G. Mostofa et al., (2015) reported Hydrogen Sulfide Regulates Salt Tolerance in Rice by Maintaining Na+/K+ Balance, Mineral Homeostasis and Oxidative Metabolism Under Excessive Salt Stress. Front. Plant Sci. 6:1055. doi: 10.3389/fpls.2015.01055.

15. K. Miah et al., (2015). Agrobacterium Mediated Genetic Transformation of Rice for Salinity Tolerance for Combating Climate Change. Volume 8, Issue 3 Ver. I, PP 28-35.

16. Zeba I. Seraj et al.,(2015). Salinity and drought tolerance conferred by in planta transformation of SNAC1 transcription factor into a high-yielding rice variety of Bangladesh. Acta physiologiae plantarum 37(4), DOI: 10.1007/s11738-015-1817-8.

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LITERATURE REVIEW ON SALT RESISTANT TRANSGENIC RICE VARIETY IN BANGLADESH

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