Interspecific divergence of circadian properties in duckweed plants

The circadian clock system is widely conserved in plants; however, divergence in circadian rhythm properties is poorly understood. We conducted a comparative analysis of the circadian properties of closely related duckweed species. Using a particle bombardment method, a circadian bioluminescent reporter was introduced into duckweed plants. We measured bioluminescence circadian rhythms of eight species of the genus Lemna and seven species of the genus Wolffiella at various temperatures (20, 25, and 30 °C) and light conditions (constant light or constant dark). Wolffiella species inhabit relatively warm areas and lack some tissues/organs found in Lemna species. Lemna species tended to show robust bioluminescence circadian rhythms under all conditions, while Wolffiella species showed lower rhythm stability, especially at higher temperatures. For Lemna, two species (L. valdiviana and L. minuta) forming a clade showed relatively lower circadian stability. For Wolffiella, two species (W. hyalina and W. repanda) forming a clade showed extremely long period lengths. The circadian properties of species primarily reflect their phylogenetic positions. The relationships between geographical and morphological factors and circadian properties are also suggested.

In planta genome editing with CRISPR/Cas9 ribonucleoproteins

In planta genome editing represents an attractive approach to engineering crops/varieties that are recalcitrant to culture-based transformation methods. Here, we report the direct delivery of CRISPR/Cas9 ribonucleoproteins into the shoot apical meristem using in planta particle bombardment and introduction of a semidwarf1 (sd1)-orthologous mutation into wheat. The triple knockout tasd1 mutant of an elite wheat variety reduced culm length by 10% without a reduction in yield.

Identification of Resistance Genes to Phytophthora sojae in Domestic Soybean Cultivars from China Using Particle Bombardment

Phytophthora root and stem rot caused by Phytophthora sojae is a destructive disease that afflicts soybean plants throughout the world. The use of resistant soybean cultivars is the primary means of managing this disease, as well as the most effective and economical approach. There are abundant soybean germplasm resources in China that could be deployed for breeding programs; however, the resistance genes (Rps genes) in most cultivars are unknown, leading to uncertainty concerning which are resistant cultivars for use. The resistance genes Rps1a, Rps1c, and Rps1k prevent root and stem rot caused by most P. sojae isolates within a Chinese field population. This study identified three Rps genes in Chinese domestic soybean cultivars using three related avirulence genes by particle bombardment. The complex genetic diversity of soybean cultivars and P. sojae strains has made it difficult to define single Rps genes without molecular involvement. Gene cobombardment is a method for identifying Rps genes quickly and specifically. We showed that cultivars Dongnong 60 and Henong 72 contained Rps1a, while Hedou 19, Henong 76, 75-3, Wandou 21020, Zheng 196, Wandou 28, Heinong 71, and Wandou 29 all contained Rps1c. The cultivars Jidou 12, Henong 72, Heinong 71, and Wandou 29 contained Rps1k. The cultivar Henong 72 contained both Rps1a and Rps1k, while Wandou 29 and Heinong 71 contained both Rps1c and Rps1k. We then evaluated the phenotype of 11 domestic soybean cultivars reacting to P. sojae using the isolates P6497 and Ps1. The 11 domestic cultivars were all resistant to P6497 and Ps1. This research provides source materials and parent plant strains containing Rps1a, Rps1c, and Rps1k for soybean breeding programs.