NAC transcription factors are one of the largest transcription factor families having functions in a variety of stress responses. Few NACs have been reported for interactions between wheat and the wheat rust fungus Puccinia triticina (Pt). In this study, based on analysis of RNA-seq data from wheat line TcLr19 inoculated by Pt, the NAC transcription factor TaNAC069 was cloned from wheat, and its transcriptional activity and homologous dimer formation were verified. Quantitative real-time PCR analysis showed that the expression of TaNAC069 was induced by Pt and associated signaling molecules. To further characterize the function of the TaNAC069 gene in wheat resistance to Pt, virus-induced gene silencing (VIGS) was utilized, and it revealed that Pt resistance in TaNAC069-silenced plants was significantly reduced. Potential interaction targets of TaNAC069 from wheat and Pt were screened and identified by yeast two-hybrid technology. Eukaryotic elongation factor eEF1A, CBSX3 protein, and cold acclimation protein WCOR410c were screened by yeast one-hybrid technology. The results indicate that the TaNAC069 gene plays a positive regulatory role in wheat resistance to Pt, laying a good foundation to analyze the molecular mechanisms of TaNAC069 and its functional role in wheat resistance to Pt.

Background The casein kinase 2 (CK2), one of the major multifunctional protein kinases in plant cells, is expressed ubiquitously and is essential for survival in response to different stresses. However, the role of CK2 in the interaction between wheat and Puccinia triticina (Pt) is still unknown. Results Our earlier studies have found that one casein kinase CK2 named TaCK2α was screened from RNA-seq data in wheat line TcLr19 inoculated by Pt. In this study, quantitative real-time PCR (qPCR) analysis showed that TaCK2αexpression was induced by Pt race PHNT and signaling molecules, suggesting that it might play a role in the interaction of wheat-Pt associated with abscisic acid (ABA) and ethylene (ETH) signaling molecules. To further determine the function of the TaCK2αgene in wheat resistance to Pt, protein-mediated phenotyping assays in vivo and antifungal activity assays in vitrodemonstrated that wheat leaves infiltrated with pure TaCK2α protein developed significantly less disease compared to control leaves. Then virus-induced gene silencing (VIGS) system confirmed that TaCK2α was involved in regulating wheat resistance to Pt. In addition, 24 candidates with potential interacting with TaCK2α (2 disease resistance related protein, 11 stress resistance related protein, 1 photosynthesis related proteins, and 10 functional indeterminate protein) were selected in wheat by pull-down assay combination with mass spectrometry (MS). Conclusions Overall, our results indicate that TaCK2α gene mediated wheat resistance to Pt, laying a good foundation to analyze the molecular mechanisms of CK2 in plant resistance to biotic stress.

Peach (Prunus persica [L.] Batsch) as an economically important fruit tree is widely cultivated in Shenzhou, China. In September 2021, peach rust was observed in the peach tree in Shenzhou City, Hebei Province (lat. 38°02'56'' N, long. 115°54'57'' E, altitude 22 m). We investigated a peach orchard with a planting area of 1334 m2, where a total of 162 peach trees were planted, and found that about 10% of peach trees exhibited severe disease symptoms. The leaves of infected plant developed 100% disease symptoms, in which 50% of the infected leaves showed about 10 small pale-yellow spots on the front of leaves and reddish-brown pustules on the corresponding abaxial surface of leaves. Urediniospores varied from obovoid to clavate in shape, sometimes in irregular shape. They were orange-brown, echinulate near base with spines smaller towards apex and often smooth at apex, with germ pores 3-4 at equator, size ranging from 25.4 to 38.6 × 10.1 to 18.7 µm (n=100), and with wall 1 to 1.5μm thick at sides and 5-7 μm thick at apex. Golden capitate paraphyses were present, ranging from 25 to 40 µm in length, with a head in diameter of 12 to 14 µm and a tail in width of 5.2 to 6.5 µm. Based on the rust morphological characters, this pathogen was primarily identified as Tranzschelia discolor (Fuckel) Tranzschel & Litv. (Hiratsuka et al. 1992). For molecular identification, total DNA was extracted from 2 isolates, respectively, and the internal transcribed spacer (ITS) region was PCR-amplified using the primer set ITS5-u and ITS4-u (Pfunder et al. 2001). Obtained sequences were compared with sequences in the GenBank repository using BLAST algorithm. BLAST showed a 100% sequence identify to T. discolor (accession nos. AB097449、MT786217、KU712078、KY764179、MH599069). The sequence has been deposited in GenBank with (accession NO. ON950745 and ON950747). Thus, combining morphological observations and molecular identification, the isolate was identified as T. discolor. The pathogenicity was verified by inoculating the abaxial surface of peach leaves with a suspension of 1 × 106 urediniospores/ml. Peach leaves sprayed with sterile water were used as controls. The inoculated peach trees were placed in a greenhouse at 20°C under dark for 24 hours and maintained at 100% relative humidity to promote disease development. Next, the peach trees were grown in a greenhouse at 20°C with a 12 h day length and symptoms were observed on the leaves 14 days after inoculation. In contrast, the control leaves were asymptomatic. Previous studies reported that peach rust occurred in Oman, Korea and Brazil was caused by T. discolor. (Deadman M L, et al.2007, Shin, H D, et al. 2019, Vidal G S, et al. 2021). To our knowledge, this is the first report of T. discolor as a causal agent causing peach leaf rust in Northern China, which will enable us to rapidly diagnose this disease, identify the occurrence of this disease and develop adequate management strategies to control it in China.