Vacuolar H⁺-pumping pyrophosphatases (VPs) provide a proton gradient for Na⁺ sequestration in the tonoplast; however, the regulatory mechanisms of VPs in developing salt tolerance have not been fully elucidated. Here, we cloned a barley (Hordeum vulgare) VP gene (HVP10) that was identified previously as the HvNax3 gene. Homology analysis showed VP10 in plants had conserved structure and sequence and likely originated from the ancestors of the Ceramiales order of Rhodophyta (Cyanidioschyzon merolae). HVP10 was mainly expressed in roots and upregulated in response to salt stress. After salt treatment for 3 weeks, HVP10 knockdown (RNA interference) and knockout (CRISPR/Cas9 gene editing) barley plants showed greatly inhibited growth and higher shoot Na⁺ concentration, Na⁺ transportation rate and xylem Na⁺ loading relative to wild-type (WT) plants. Reverse transcription quantitative polymerase chain reaction and microelectronic Ion Flux Estimation results indicated that HVP10 likely modulates Na⁺ sequestration into the root vacuole by acting synergistically with Na⁺/H⁺ antiporters (HvNHX1 and HvNHX4) to enhance H⁺ efflux and K⁺ maintenance in roots. Moreover, transgenic rice (Oryza sativa) lines overexpressing HVP10 also showed higher salt tolerance than the WT at both seedling and adult stages with less Na⁺ translocation to shoots and higher grain yields under salt stress. This study reveals the molecular mechanism of HVP10 underlying salt tolerance and highlights its potential in improving crop salt tolerance.

Indica-japonica hybrid rice cultivars show great yield potential but poor eating quality and require more nitrogen (N) input relative to japonica rice. However, the effect of N levels on the eating quality of indica-japonica hybrid rice is little known. A field experiment was carried out to investigate the effects of four N levels on two indica-japonica hybrid rice cultivars (Yongyou12 and Yongyou17) differing in eating quality. The results showed that the contents of amylose chains and water-insoluble storage proteins, especially prolamin, increased largely under a high N level, leading to deterioration of the rice-eating quality, although a low N level (100 N kg/ha) had a less negative effect on the eating quality. Moreover, both of the indica-japonica hybrids had high ratios of inferior grains (IG), and the ratio of IG increased with the N level. Grain weight and the immature ratio of IG were reduced and increased with the N level, respectively, which are also factors for deterioration of the eating quality. The two cultivars differed greatly in the responses of eating quality to the N level, with Yongyou17 being more sensitive than Yongyou12. The current results indicated that a high N level deteriorates the eating quality of indica-japonica hybrid rice mainly due to a large increase of IG.

Plants are frequently exposed to various abiotic stresses, including aluminum, cadmium and salinity stress. Barley (Hordeum vulgare) displays wide genetic diversity in its tolerance to various abiotic stresses. In this study, small RNA and degradome libraries from the roots of a barley cultivar, Golden Promise, treated with aluminum, cadmium and salt or controls were constructed to understand the molecular mechanisms of microRNAs in regulating tolerance to these stresses. A total of 525 microRNAs including 198 known and 327 novel members were identified through high-throughput sequencing. Among these, 31 microRNAs in 17 families were responsive to these stresses, and Gene Ontology (GO) analysis revealed that their targeting genes were mostly highlighted as transcription factors. Furthermore, five (miR166a, miR166a-3p, miR167b-5p, miR172b-3p and miR390), four (MIR159a, miR160a, miR172b-5p and miR393) and three (miR156a, miR156d and miR171a-3p) microRNAs were specifically responsive to aluminum, cadmium and salt stress, respectively. Six miRNAs, i.e., miR156b, miR166a-5p, miR169a, miR171a-5p, miR394 and miR396e, were involved in the responses to the three stresses, with different expression patterns. A model of microRNAs responding to aluminum, cadmium and salt stresses was proposed, which may be helpful in comprehensively understanding the mechanisms of microRNAs in regulating stress tolerance in barley.