An NLR paralog Pit2 generated from tandem duplication of Pit1 fine-tunes Pit1 localization and function.

NLR family proteins act as intracellular receptors. Gene duplication amplifies the number of NLR genes, and subsequent mutations occasionally provide modifications to the second gene that benefits immunity. However, evolutionary processes after gene duplication and functional relationships between duplicated NLRs remain largely unclear. Here, we report that the rice NLR protein Pit1 is associated with its paralogue Pit2. The two are required for the resistance to rice blast fungus but have different functions: Pit1 induces cell death, while Pit2 competitively suppresses Pit1-mediated cell death. During evolution, the suppression of Pit1 by Pit2 was probably generated through positive selection on two fate-determining residues in the NB-ARC domain of Pit2, which account for functional differences between Pit1 and Pit2. Consequently, Pit2 lost its plasma membrane localization but acquired a new function to interfere with Pit1 in the cytosol. These findings illuminate the evolutionary trajectory of tandemly duplicated NLR genes after gene duplication.

Seed Halopriming: A Promising Strategy to Induce Salt Tolerance in Indonesian Pigmented Rice.

Unfavorable environmental conditions and climate change impose stress on plants, causing yield losses worldwide. The Indonesian pigmented rice (Oryza sativa L.) cultivars Cempo Ireng Pendek (black rice) and Merah Kalimantan Selatan (red rice) are becoming popular functional foods due to their high anthocyanin contents and have great potential for widespread cultivation. However, their ability to grow on marginal, high-salinity lands is limited. In this study, we investigated whether seed halopriming enhances salt tolerance in the two pigmented rice cultivars. The non-pigmented cultivars IR64, a salt-stress-sensitive cultivar, and INPARI 35, a salt tolerant, were used as control. We pre-treated seeds with a halopriming solution before germination and then exposed the plants to a salt stress of 150 mM NaCl at 21 days after germination using a hydroponic system in a greenhouse. Halopriming was able to mitigate the negative effects of salinity on plant growth, including suppressing reactive oxygen species accumulation, increasing the membrane stability index (up to two-fold), and maintaining photosynthetic pigment contents. Halopriming had different effects on the accumulation of proline, in different rice varieties: the proline content increased in IR64 and Cempo Ireng Pendek but decreased in INPARI 35 and Merah Kalimantan Selatan. Halopriming also had disparate effects in the expression of stress-related genes: OsMYB91 expression was positively correlated with salt treatment, whereas OsWRKY42 and OsWRKY70 expression was negatively correlated with this treatment. These findings highlighted the potential benefits of halopriming in salt-affected agro-ecosystems.

Identification and characterization of drought-tolerant local pigmented rice from Indonesia.

Water is essential to support life. Because limited water availability may affect their life cycles, plants have developed multiple responses to drought stress. Plant physiological and metabolic changes during drought may reflect changes that occur at the level of gene expression. In this study, we investigated the variation in drought-mitigating strategies employed by pigmented rice (Oryza sativa) varieties and the genes involved in their possible drought tolerance. We screened 21 local pigmented rice cultivars from Indonesia for increased drought tolerance using the fraction transpirable soil water method to exert precise control of the drought stress imposed on plants. We then determined the expression of OsDREB1A, OsNAC6, OsNHX1, OsCuZnSOD2, OsOSCAT2, and OsCAT3 in plants grown under well-watered conditions and under moderate or severe drought stress. Among the pigmented rice cultivars, Merah Pari Eja had the greatest drought tolerance, while the red rice Inpari 24 had the highest mortality rate (60%). We also included the white rice cultivar Putih Payo, which is fully sensitive to drought (with 100% mortality under the conditions used) as a negative control. Gene expression profiling revealed a general upregulation of drought-related genes in Merah Pari Eja and a downregulation of such genes in the other two cultivars. Measurements of antioxidant enzyme activity, leaf damage, free radicals, chlorophyll, and anthocyanin contents provided further evidence that Merah Pari Eja is more drought tolerant than the other two cultivars. We conclude that OsDREB1A, OsNAC6, OsNHX1, OsCuZnSOD2, OsOSCAT2 and OsCAT3 expression patterns can reveal plants that have increased drought tolerance.