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MADS-box proteins, a well-conserved family of transcription factors in eukaryotic organisms, specifically regulate a wide range of cellular functions, including primary metabolism, cell cycle, and cell identity. However, little is known about roles of the MADS-box protein family in the fungal pathogen Sclerotinia sclerotiorum. In this research, the S. sclerotiorum MADS-box gene SsMADS was cloned; it encodes a protein that is highly similar to Mcm1 orthologs from Saccharomyces cerevisiae and other fungi, and includes a highly conserved DNA-binding domain. MADS is a member of the MADS box protein SRF (serum response factor) lineage. SsMADS function was investigated using RNA interference. Silenced strains were obtained using genetic transformation of the RNA interference vectors pS1-SsMADS and pSD-SsMADS. SsMADS expression levels in silenced strains were analyzed using RT-PCR. The results showed that SsMADS mRNA expression in these silenced strains was reduced to different degrees, and growth rate in these silenced strains was significantly decreased. Infecting tomato leaflets with silenced strains indicated that SsMADS was required for leaf pathogenesis in a susceptible host. Our results suggest that the MADS-box transcription factor SsMADS is involved in S. sclerotiorum growth and virulence.

The jacalin-related lectins (JRLs) are widely distributed in plants and are involved in plant development and multiple stress responses. However, the characteristics of the HvJRL gene family at the genome-wide level and the roles of JRLs in barley’s response to low-nitrogen (LN) stress have been rarely reported. In this study, 32 HvJRL genes were identified and unevenly distributed at both ends of the seven chromosomes in barley. HvJRL proteins generally exhibited low sequence similarity but shared conserved jacalin domains by multiple sequence analysis. These proteins were classified into seven subfamilies based on phylogenetic analysis, with a similar gene structure and conserved motifs in the same subfamily. The HvJRL promoters contained a large number of diverse cis-elements associated with hormonal response and stress regulation. Based on the phylogenetic relationships and functionally known JRL homologs, it was predicted that some HvJRLs have the potential to serve functions in multiple stress responses but not nutrition deficiency stress. Subsequently, nine differentially expressed genes (DEGs) encoding eight HvJRL proteins were identified in two barley genotypes with different LN tolerance by transcriptome analysis. Furthermore, 35S:HvHorcH transgenic Arabidopsis seedlings did enhance LN tolerance, which indicated that HvHorcH may be an important regulator of LN stress response (LNSR). The HvJRL DEGs identified herein could provide new candidate genes for LN tolerance studies.

Improvement of low nitrogen (LN) tolerance or nitrogen use efficiency (NUE) in crops is imperative for environment-friendly agriculture development. The basic helix-loop-helix (bHLH) transcription factors are involved in multiple abiotic stresses and are suitable as candidate genes for improving LN tolerance. Few studies were performed on the characterization of the HvbHLH gene family and their function in response to LN stress in barley. In this study, 103 HvbHLH genes were identified through genome-wide analysis. HvbHLH proteins were classified into 20 subfamilies based on phylogenetic analysis in barley, which was supported by conserved motifs and gene structure analysis. The stress-related cis-element analysis in the promoters showed that HvbHLHs are probably involved in multiple stress responses. By phylogenetic analysis of HvbHLHs and bHLHs in other plants, some HvbHLHs were predicted to play roles in response to nutrition deficiency stress. Furthermore, at least 16 HvbHLHs were differentially expressed in two barley genotypes differing in LN tolerance under LN stress. Finally, overexpression of HvbHLH56 enhanced LN stress tolerance in transgenic Arabidopsis, suggesting it is an important regulator in LN stress response. The differentially expressed HvbHLHs identified herein may be valuable for the breeding of barley cultivars with LN tolerance.

Broussonetia papyrifera has a high lignocellulose content leading to poor palatability and low digestion rate of ruminants. Thus, dynamic profiles of fermentation lignocellulose characteristics, microbial community structure, potential function, and interspecific relationships of B. papyrifera mixing with wheat bran in different ratios: 100:0 (BP100), 90:10 (BP90), 80:20 (BP80), and 65:35 (BP65) were investigated on ensiling days 5, 15, 30, and 50. The results showed that adding bran increased the degradation rate of hemicellulose, neutral detergent fiber, and the activities of filter paper cellulase, endoglucanase, acid protease, and neutral protease, especially in the ratio of 65:35. Lactobacillus, Pediococcus, and Weissella genus bacteria were the dominant genera in silage fermentation, and Pediococcus and Weissella genus bacteria regulated the process of silage fermentation. Compared with monospecific B. papyrifera silage, adding bran significantly increased the abundance of Weissella sp., and improved bacterial fermentation potential in BP65 (p < 0.05). Distance-based redundancy analysis showed that lactic acid bacteria (LAB) were significantly positive correlated with most lignocellulose content and degrading enzymes activities, while Monascus sp. and Syncephalastrum sp. were opposite (p < 0.05). Co-occurrence network analysis indicated that there were significant differences in microbial networks among different mixing ratios of B. papyrifera silage prepared with bran. There was a more complex, highly diverse and less competitive co-occurrence network in BP65, which was helpful to silage fermentation. In conclusion, B. papyrifera ensiled with bran improved the microbial community structure and the interspecific relationship and reduced the content of lignocellulose.

Salinity is one of the limiting factors of wheat production worldwide. A total of 334 internationally derived wheat genotypes were employed to identify new germplasm resources for salt tolerance breeding. Salt stress caused 39, 49, 58, 55, 21 and 39% reductions in shoot dry weight (SDW), root dry weight (RDW), shoot fresh weight (SFW), root fresh weight (RFW), shoot height (SH) and root length (RL) of wheat, respectively, compared with the control condition at the seedling stage. The wheat genotypes showed a wide genetic and tissue diversity for the determined characteristics in response to salt stress. Finally, 12 wheat genotypes were identified as salt-tolerant through a combination of one-factor (more emphasis on the biomass yield) and multifactor analysis. In general, greater accumulation of osmotic substances, efficient use of soluble sugars, lower Na+/K+ and a higher-efficiency antioxidative system contribute to better growth in the tolerant genotypes under salt stress. In other words, the tolerant genotypes are capable of maintaining stable osmotic potential and ion and redox homeostasis and providing more energy and materials for root growth. The identified genotypes with higher salt tolerance could be useful for developing new salt-tolerant wheat cultivars as well as in further studies to underline the genetic mechanisms of salt tolerance in wheat.