No abstract

Salinity and drought severely affect both plant growth and productivity, making the isolation and characterization of salinity- or drought-inducible promoters suitable for genetic improvement of crop resistance highly desirable. In this study, a 1468-bp sequence upstream of the translation initiation codon ATG of the promoter for ZmGAPP (maize Type-II H+-pyrophosphatase gene) was cloned. Nine 5´ deletion fragments (D1–D9) of different lengths of the ZmGAPP promoter were fused with the GUS reporter and translocated into tobacco. The deletion analysis showed that fragments D1–D8 responded well to NaCl and PEG stresses, whereas fragment D9 and CaMV 35S did not. The D8 segment (219 bp; -219 to -1 bp) exhibited the highest promoter activity of all tissues, with the exception of petals among the D1–D9 transgenic tobacco, which corresponds to about 10% and 25% of CaMV 35S under normal and NaCl or PEG stress conditions, respectively. As such, the D8 segment may confer strong gene expression in a salinity and osmotic stress inducible manner. A 71-bp segment (-219 to -148 bp) was considered as the key region regulating ZmGAPP response to NaCl or PEG stress, as transient transformation assays demonstrated that the 71-bp sequence was sufficient for the salinity or osmotic stress response. These results enhance our understanding of the molecular mechanisms regulating ZmGAPP expression, and that the D8 promoter would be an ideal candidate for moderating expression of drought and salinity response genes in transgenic plants.

Salinity and drought often affect plant growth and crop yields. Cloning and identification of salinity and drought stress inducible promoters is of great significance for their use in the genetic improvement of crop resistance. Previous studies showed that phosphatidylinositol synthase is involved in plant salinity and drought stress responses but its promoter has not been characterized by far. In the study, the promoter (pZmPIS, 1834 bp upstream region of the translation initiation site) was isolated from maize genome. To functionally validate the promoter, eight 5′ deletion fragments of pZmPIS in different lengths were fused to GUS to produce pZmPIS::GUS constructs and transformed into tobacco, namely PZ1–PZ8. The transcription activity and expression pattern obviously changed when the promoter was truncated. Previous studies have demonstrated that NaCl and PEG treatments are usually used to simulate salinity and drought treatments. The results showed that PZ1–PZ7 can respond well upon NaCl and PEG treatments, while PZ8 not. PZ7 (467 bp) displayed the highest transcription activity in all tissues of transgenic tobacco amongst 5′ deleted promoter fragments, which corresponds to about 20 and 50% of CaMV35S under normal and NaCl or PEG treatment, respectively. This implied that PZ7 is the core region of pZmPIS which confers high-level gene expression and NaCl or PEG inducible nature. The 113 bp segment between PZ7 and PZ8 (-467 to -355 bp) was considered as the key sequence for ZmPIS responding to NaCl or PEG treatment. GUS transient assay in tobacco leaves showed that this segment was sufficient for the NaCl or PEG stress response. Bioinformatic analysis revealed that the 113 bp sequence may contain new elements that are crucial for ZmPIS response to NaCl or PEG stress. These results promote our understanding on transcriptional regulation mechanism of ZmPIS and the characterized PZ7 promoter fragment would be an ideal candidate for the overexpression of drought and salinity responsive gene to improve crop resistance.

The effects of nitrification inhibitor in tea gardens with different urea–N rates have rarely been assessed. For eight months, a glasshouse experiment was conducted to investigate the effects of a nitrification inhibitor (3, 4–dimethylpyrazole phosphate, DMPP) on the changes of soil pH and inorganic N loss. Urea (0, 300, 500, and 800 kg N ha−1) with or without DMPP (1% of urea–N applied) were added to pots that hosted six plants that were three years old. Next, three leaching events were conducted with 600 mL of water after 7, 35, and 71 days of intervention while soil samples were collected to determine pH and inorganic N. Averaged across sampling dates, urea–N application at an increasing rate reduced soil pH with the lowest values at 800 kg urea–N ha−1. Adding DMPP increased soil pH up to a rate of 500 kg ha−1. Irrespective of the addition of DMPP, gradient urea–N application increased the leaching loss of inorganic N. On overage, DMPP increased soil pH and decreased leaching losses of total inorganic N, suggesting a higher soil N retention. Therefore, we believe that this increase in soil pH is associated with a relatively lower proton release from the reduced nitrification in the DMPP–receiving pots. This nitrification reduction also contributed to the N loss reduction (NO3−–N). Altogether, our results suggest that DMPP can reduce N leaching loss while maintaining the pH of tea–cultivated soils. Therefore, DMPP application has a significant potential for the sustainable N management of tea gardens.

In order to understand the response of nitrate metabolism in seedlings of oilseed rape (Brassica napus L.) to low oxygen stress (LOS), two cultivars were studied at different light, LOS time and exogenous nitrate concentration under hydroponic stress. Results show that N-uptake and dry matter of rape seedlings were decreased after LOS stress while nitrate accumulation (NA) under LOS was induced by darkness. Nitrate accumulation peaked at 3d while root activity (RA, defined as dehydrogenase activity) decreased with prolonged waterlogging exposure. Exogenous nitrate significantly elevated NA and RA. Tungstate (TS) and LOS inhibited nitrate reductase (NR) activity while NR transcription and activity were enhanced by exogenous nitrate. Low oxygen stress stimulated the activity of superoxide dismutase (SOD) and peroxidase (POD) slightly, but inhibited that of catalase (CAT). B. napus L. Zhongshuang 10 (ZS10), a LOS tolerant cultivar, displayed smaller decrease upon dry matter under LOS, higher NA in darkness and lower NA in light than B. napus L. Ganlan CC (GAC), a LOS sensitive variety. ZS10 had lower NA and higher RA after waterlogging and exogenous nitrate treatment, and higher NR activity under TS inhibition than GAC, but the activity of antioxidant enzymes did not change under LOS. The results indicate that nitrate metabolism involved tolerance of rape seedlings to LOS, with lower accumulation and higher reduction of nitrate being related to higher LOS tolerance of rape seedlings exposed to waterlogging.