Abstract
Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas.
Highlights
Cumin (Cuminum cyminum L.) is an aromatic medicinal herb; dried seeds are consumed as spices, and the plant is well documented with various functional and nutritional properties in the scientific literature
The plant attains a height of 30–40 cm, thrives well in tropical regions because of the fibrous root, and growth period coincides with winter and springs rainfall that provides drought tolerance ability [6,7, 10]
Productivity and seed yield of cumin is highly susceptible to soil salinization i.e. salinity stress which is irresistible process in semi-arid regions due to natural occurrence of high amount of soluble salt [11,12], uneven rainfall distribution and poor quality water for irrigational purposes leading to excess salt deposition in the root zone of growing plants [7, 13,14]
Summary
Cumin (Cuminum cyminum L.) is an aromatic medicinal herb; dried seeds are consumed as spices, and the plant is well documented with various functional and nutritional properties in the scientific literature. Transgenic Cumin with Salt Stress Tolerance plant parts such as seed, fruit and flower are used for flavor enhancement and as food preservatives [1,2]. Productivity and seed yield of cumin is highly susceptible to soil salinization i.e. salinity stress which is irresistible process in semi-arid regions due to natural occurrence of high amount of soluble salt [11,12], uneven rainfall distribution and poor quality water for irrigational purposes leading to excess salt deposition in the root zone of growing plants [7, 13,14]. The germination of seed, seedling growth and other metabolic activities of this aromatic plant is severely affected by salinity stress which further decreases umbel formation and seed harvest index causing yield and productivity loss of cumin seeds [5, 15,16]. The present scenario of agricultural land highly threatens to productivity loss, as about 20% (62 million hectares) of irrigated land has already subjected to salt-induced degradation [17]
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