Drought resistance improvement in rice: An integrated genetic and resource management strategy

Drought is the major constraint to rice production in rainfed areas across Asia and sub-Saharan Africa. In the context of current and predicted water scarcity, increasing irrigation is generally not a viable option for alleviating drought problems in rainfed rice-growing systems. It is therefore critical that genetic management strategies for drought focus on maximum extraction of available soil moisture and its efficient use in crop establishment and growth to maximize biomass and yield. Extensive genetic variation for drought resistance exists in rice germplasm. However, the current challenge is to decipher the complexities of drought resistance in rice and exploit all available genetic resources to produce rice varieties combining drought adaptation with high yield potential, quality, and resistance to biotic stresses. The strategy described here aims at developing a pipeline for elite breeding lines and hybrids that can be integrated with efficient management practices and delivered to rice farmers. This involves the development of high-throughput, high-precision phenotyping systems to allow genes for yield components under stress to be efficiently mapped and their effects assessed on a range of drought-related traits, and then moving the most promising genes into widely grown rice mega-varieties, while scaling up gene detection and delivery for use in marker-aided breeding.Rachid Serraj, Kenneth L. McNally, Inez Slamet-Loedin, Ajay Kohli, Stephan M. Haefele, Gary Atlin and Arvind Kuma

Large-scale production and evaluation of marker-free indica rice IR64 expressing phytoferritin genes

Biofortification of rice (Oryza sativa L.) using a transgenic approach to increase the amount of iron in the grain is proposed as a low-cost, reliable, and sustainable solution to help developing countries combat anemia. In this study, we generated and evaluated a large number of rice or soybean ferritin over-accumulators in rice mega-variety IR64, including marker-free events, by introducing soybean or rice ferritin genes into the endosperm for product development. Accumulation of the protein was confirmed by ELISA, in situ immunological detection, and Western blotting. As much as a 37- and 19-fold increase in the expression of ferritin gene in single and co-transformed plants, respectively, and a 3.4-fold increase in Fe content in the grain over the IR64 wild type was achieved using this approach. Agronomic characteristics of a total of 1,860 progenies from 58 IR64 single independent transgenic events and 768 progenies from 27 marker-free transgenic events were evaluated and most trait characteristics did not show a penalty. Grain quality evaluation of high-Fe IR64 transgenic events showed quality similar to that of the wild-type IR64. To understand the effect of transgenes on iron homeostasis, transcript analysis was conducted on a subset of genes involved in iron uptake and loading. Gene expression of the exogenous ferritin gene in grain correlates with protein accumulation and iron concentration. The expression of NAS2 and NAS3 metal transporters increased during the grain milky stage

Broad-spectrum resistance to bacterial blight in rice using genome editing

Bacterial blight of rice is an important disease in Asia and Africa. The pathogen, Xanthomonas oryzae pv. oryzae (Xoo), secretes one or more of six known transcription-activator-like effectors (TALes) that bind specific promoter sequences and induce, at minimum, one of the three host sucrose transporter genes SWEET11, SWEET13 and SWEET14, the expression of which is required for disease susceptibility. We used CRISPR-Cas9-mediated genome editing to introduce mutations in all three SWEET gene promoters. Editing was further informed by sequence analyses of TALe genes in 63 Xoo strains, which revealed multiple TALe variants for SWEET13 alleles. Mutations were also created in SWEET14, which is also targeted by two TALes from an African Xoo lineage. A total of five promoter mutations were simultaneously introduced into the rice line Kitaake and the elite mega varieties IR64 and Ciherang-Sub1. Paddy trials showed that genome-edited SWEET promoters endow rice lines with robust, broad-spectrum resistance