Overexpression of annexin gene in rice ( Oryza sativa L.) for salinity and water stress
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PLANT TISSUE CULTURE
Overexpression of annexin gene in rice (Oryza sativa L.) for salinity and water stress Subhadra Rani Mallick 1 & Kundansing Rajpalsing Jadhao 1 & Gyana Ranjan Rout 1 Received: 30 April 2020 / Accepted: 20 July 2020 / Editor: Prakash Lakshmanan # The Society for In Vitro Biology 2020
Abstract The aim of the study is to develop transgenic rice with annexin genes (AnnBj2) to determine its salinity and water stress tolerance through Agrobacterium-mediated genetic transformation. The transgenic somatic embryos were developed on Murashige and Skoog medium fortified with 0.5 mg L−1 kinetin and 3.0 mg L−1 2,4-D. About 80% of the transformed somatic embryos germinated and was successfully established in the transgenic greenhouse. The transformed plantlets (T0 and T1 generation) were identified through PCR using NPT-II and AnnBj2 gene–specific primers as well as positive uidA reporter gene expression. Compared with non-transformed plants, the transgenic rice plants overexpressing AnnBj2 gene exhibited salt tolerance at the seedling stage. Seeds generated at T0 and T1 generations were further studied to determine their salinity tolerance through nutrient culture and pot culture experiments for water stress. When compared with the non-transgenic control, transgenic seeds (T0 and T1) had increased germination on a nutrient medium having 200 mM NaCl. The data showed that the shoot and root lengths were longer at 200 mM NaCl when compared with those of the non-transformed plants. Further, the biochemical assessment (photosynthetic pigment analysis, protein, proline content, and oxidative enzyme activity) was performed between transformed (T0 and T1) and non-transformed plantlets under stress. It was noted that the transformed plantlets were higher in chlorophyll and proline content as well as oxidative enzyme activity as compared with the non-transformed plants. Keywords Genetic transformation . Indica rice . Somatic embryogenesis . Water stress . Salinity stress
Introduction Rice (Oryza sativa L.) is one of the important staple foods consumed by more than three billion people worldwide, and people from Asian countries cultivate and consume more than 90% of available rice (IRRI 2001). About 60% of the world’s population depends on rice in daily life. It is cultivated under different agroclimatic regions and various ecologies including irrigated and rainfed upland and lowland, and deep water. Biotic and abiotic stresses affect crop production to a great extent (Wahab 2017; Bzour et al. 2018). Due to industrialization and urbanization, the fertile land for rice cultivation has been reduced and it is a great challenge to provide food to the population (Kumar 2017). Progress in genetic transformation of rice through Agrobacterium mediation has been * Gyana Ranjan Rout [email protected] 1
Department of Agricultural Biotechnology, College of Agriculture, Odisha University of Agriculture and Technology, Bhubaneswar, Odisha 751003, India
standardized by several researchers (Raineri et al. 1990; Chan et al. 1
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