Development of an inoculation technique for rapidly evaluating maize inbred lines for resistance to stalk rot caused by Fusarium spp. in the field

Maize (Zea mays L.) stalk rot, caused primarily by the soil-borne fungal pathogen Fusarium spp., reduces maize quality and yield worldwide. This study was undertaken to develop and utilize a rapid continuous injection inoculation technique to evaluate maize inbred lines for resistance to Fusarium spp. under field conditions, which could facilitate the identification and development of new sources of host resistance to manage the disease. Continuous injection inoculation is a rapid, stable, and simple method that can evaluate the resistance of maize inbred lines to Fusarium stalk rot (FSR) within 20 days. To verify the feasibility and reliability of inoculation method, Fusarium graminearum, F. proliferatum, and F. subglutinans were isolated, identified, and inoculated into maize at the six-leaf stage (V6) by a veterinary adjustable bottle continuous vaccination syringe. Our results showed that out of a total of 97 inbred maize lines, six (6.2%) showed high resistance to maize stalk rot, 20 showed resistance (20.6%), 32 were susceptible (33.0%), and 39 were very susceptible (40.2%). Based on simple sequence repeat (SSR) markers, an analysis of molecular variance indicated a significant correlation between population of the inbred maize line and resistance to FSR (P = 0.001). Overall, this study provided a systematic, rapid, stable, and simple identification method for maize inbred lines resistant to FSR in the field. At the same time, this method was also suitable for genetic diversity analysis of maize inbred lines resistant to FSR.

Transcriptomic Analysis Revealed the Common and Divergent Responses of Maize Seedling Leaves to Cold and Heat Stresses

Temperature stresses (TS), including cold and heat stress, adversely affect the growth, development, and yield of maize (Zea mays L.). To clarify the molecular mechanisms of the tolerance of maize seedling leaves to TS, we applied transcriptomic sequencing of an inbred maize line, B73, with seedlings exposed to various temperature conditions, including normal temperature (NT, 25 °C), cold (4, 10, and 16 °C), and heat (37, 42, and 48 °C) stresses. Differentially expressed genes (DEGs) were detected in different comparison between the NT sample and each temperature-stressed sample, with 5358, 5485, 5312, 1095, 2006, and 4760 DEGs responding to TS of 4, 10, 16, 37, 42, and 48 °C, respectively. For cold and heat stresses, 189 DEGs enriched in the hydrogen peroxidase metabolic process, cellular modified amino acid metabolic process, and sulfur compound metabolic process were common. The DEGs encoding calcium signaling and reactive oxygen species scavenging enzymes demonstrated similar expression characterizations, whereas the DEGs encoding transcription factors, such as ERF, ARF, and HSF, hormone signaling, and heat shock proteins, displayed divergent expression models, implying both common and divergent responses to cold and heat stresses in maize seedling leaves. Co-expression network analysis showed that functional DEGs associated with the core regulators in response to cold and heat stresses were significantly correlated with TS, indicating their vital roles in cold and heat adaptation, respectively. Our investigation focused on the response to gradient TS, and the results presented a relatively comprehensive category of genes involved in differential TS responses. These will contribute a better understanding of the molecular mechanisms of maize seedling leaf responses to TS and provide valuable genetic resources for breeding TS tolerant varieties of maize.