Polyphosphate promotes oxidation resistance of ppk-expressing transgenic rice in low phosphorus culture.

Phosphorus (P) plays a crucial role in plant growth. Insufficient availability of inorganic phosphate (Pi) can significantly impact crop yields. To address this, we previously developed transgenic rice expressing the low polyphosphate kinase gene (ppk) - known as ETRS - to enhance the efficiency of P resource utilization. Previous studies have shown that ETRS thrives and presents high yields in the low P culture. ETRS and wild-type rice (WT) were cultivated to the heading stage at 15 µM of P in the low P (LP) culture and 300 µM of P in the normal culture (CK) to identify the molecular pathways behind low P tolerance. Our findings revealed that polyphosphate (polyP) significantly enhanced the growth performance of ETRS in the LP culture. This enhanced tolerance can be attributed to polyP's capacity to mitigate oxidative damage induced by LP. This was evidenced by the reduction in levels of superoxide radicals, hydrogen peroxide, and malondialdehyde. PolyP also improved the antioxidant capacity of ETRS under LP stress by regulating enzymatic antioxidants viz., superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as non-enzymatic antioxidants such as ascorbate (AsA) and glutathione (GSH). In addition, transcriptomics analysis suggested that polyP synthesis positively promoted the expressions of SOD, POD, and CAT related genes and played an active role in regulating the expression of AsA-GSH cycle system related genes in ETRS in the LP culture. These results strongly support the notion that polyP within ETRS mitigates oxidative damage through enhancement of the antioxidant system, ultimately bolstering tolerance to LP conditions.

The ppk-expressing transgenic rice floating bed improves P removal in slightly polluted water.

With significant economic advantages, the plant floating bed has been widely utilized in the ecological remediation of eutrophic water because of the excessive phosphorus (P) and nitrogen discharge in China. Previous research has demonstrated that polyphosphate kinase (ppk)-expressing transgenic rice (Oryza sativa L. ssp. japonica) (ETR) can increase the P absorption capacity to support rice growth and boost rice yield. In this study, the floating beds of ETR with single copy line (ETRS) and double copy line (ETRD) are built to investigate their capacity to remove aqueous P in slightly polluted water. Compared with the wild type Nipponbare (WT) floating bed, the ETR floating beds greatly reduce the total P concentration in slightly polluted water though the ETR floating beds have the same removal rates of chlorophyll-a, NO3--N, and total nitrogen in slightly polluted water. The P uptake rate of ETRD on the floating bed is 72.37% in slightly polluted water, which is higher than that of ETRS and WT on the floating beds. Polyphosphate (polyP) synthesis is a critical factor for the excessive phosphate uptake of ETR on the floating beds. The synthesis of polyP decreases the level of free intracellular phosphate (Pi) in ETR on the floating beds, simulating the phosphate starvation signaling. The OsPHR2 expression in the shoot and root of ETR on the floating bed increased, and the corresponding P metabolism gene expression in ETR was changed, which promoted Pi uptake by ETR in slightly polluted water. The Pi accumulation further promoted the growth of ETR on the floating beds. These findings highlight that the ETR floating beds, especially ETRD floating bed, have significant potential for P removal and can be exploited as a novel method for phytoremediation in slightly polluted water.

Further Mining and Characterization of miRNA Resource in Chinese Fir (Cunninghamia lanceolata).

In this study, we aimed to expand the current miRNA data bank of Chinese fir (Cunninghamia lanceolata (Lamb.) Hook.) regarding its potential value for further genetic and genomic use in this species. High-throughput small RNA sequencing successfully captured 140 miRNAs from a Chinese fir selfing family harboring vigor and depressed progeny. Strikingly, 75.7% (n = 106) of these miRNAs have not been documented previously, and most (n = 105) of them belong to the novel set with 6858 putative target genes. The new datasets were then integrated with the previous information to gain insight into miRNA genetic architecture in Chinese fir. Collectively, a relatively high proportion (62%, n = 110) of novel miRNAs were found. Furthermore, we identified one MIR536 family that has not been previously documented in this species and four overlapped miRNA families (MIR159, MIR164, MIR171_1, and MIR396) from new datasets. Regarding the stability, we calculated the secondary structure free energy and found a relatively low R2 value (R2 < 0.22) between low minimal folding free energy (MFE) of pre-miRNAs and MFE of its corresponding mature miRNAs in most datasets. When in view of the conservation aspect, the phylogenetic trees showed that MIR536 and MIR159 sequences were highly conserved in gymnosperms.

Efficient nitrate removal by Pseudomonas mendocina GL6 immobilized on biochar.

The performance of nitrate removal by Pseudomonas mendocina GL6 cells immobilized on bamboo biochar was investigated. The results showed that immobilized bacterial cells performed better nitrate removal than the free bacterial cells, and the nitrate removal rate increased from 6.51 mg/(L·h) of free cells to 8.34 mg/(L·h) of immobilized cells. The nitrate removal of immobilized bacterial cells fitted well to the zero-order kinetics model. Moreover, bath experiments showed that immobilized bacterial cells displayed more nitrate removal capacity under different conditions than free bacterial cells due to the protection of biochar carrier. The subsequent mechanistic study suggested that biochar promoted the expression level of denitrification functional genes (napA and nirK) and electron transfer genes involved in denitrification (napB and napC), which resulted in the increase of nitrate removal efficiency. Thus, biochar-immobilized P. mendocina GL6 has much potential to remove nitrate from wastewater via aerobic denitrification.

Bioaccumulations and potential human health risks assessment of heavy metals in ppk-expressing transgenic rice.

In order to reduce the phosphorus (P) resource consume, the polyphosphatekinase (ppk)-expressing transgenic rice (ETR) with high utilization efficiency of P fertilizer had been constructed. However, synthesis polyphosphates (polyP) mediated byppkin the plants have the ability of chelating heavy metals, so the potential hazards of the new elite rice variety have raised concerns. In the study, we planted ETR and wild-type Nipponbare (WT) in paddy fields in southern China. After harvest, the concentrations of eight heavy metals in rice tissues were measured, and health risks assessments were performed. The field experiment showed that the ppkexpressions were detected in the roots and straws of ETR plants but did not increase the concentrations of As, Cd, Cr, Ni and Pb in rice tissues. The Hg concentration in the ETRD root was 1.70-fold higher than that in WT, but the abundant Hg bioaccumulation in ETRD only occurred in the root. The bioaccumulation factors (BAFs) of all the detected heavy metals in the ETRS were no different from WT except for Cu and Zn. The results of human health risks assessment of heavy metals in brown rice showed that the non-carcinogenic risks of Cu or Zn in ETRD were higher than that in WT, while there was no difference in the total noncarcinogenic risk of the eight heavy metals in ETR. The carcinogenic risks of heavy metals in ETR were also comparable to that in WT. The results of this study indicated that the ppk expression in rice did not increase human health risks of heavy metals by consuming brown rice, which would provide a safety guarantee for agricultural and environmental applications of ETR not only with single-copy line but also with double-copy line.

Characterization of Aerobic Denitrifying Bacterium Pseudomonas mendocina Strain GL6 and Its Potential Application in Wastewater Treatment Plant Effluent.

To remove nitrate in wastewater treatment plant effluent, an aerobic denitrifier was newly isolated from the surface flow constructed wetland and identified as Pseudomonas mendocina strain GL6. It exhibited efficient aerobic denitrification ability, with the nitrate removal rate of 6.61 mg (N)·L-1·h-1. Sequence amplification indicated that the denitrification genes napA, nirK, norB, and nosZ were present in strain GL6. Nitrogen balance analysis revealed that approximately 74.5% of the initial nitrogen was removed as gas products. In addition, the response surface methodology experiments showed that the maximum removal of total nitrogen occurred at pH 7.76, C/N ratio of 11.2, temperature of 27.8 °C, and with shaking at 133 rpm. Furthermore, under the optimized cultivation condition, strain GL6 was added into wastewater treatment plant effluent and the removal rates of nitrate nitrogen and total nitrogen reached 95.6% and 73.6%, respectively. Thus, P. mendocina strain GL6 has high denitrification potential for deep improvement of effluent quality.

[Nutrient characteristics in leaves of broadleaved seedlings in south China].

Nutrient concentrations and contents in leaves of broadleaved Elaeocarpus sylvestris, Castanopsis hystrix, Ormosia pinnata, Michelia macclurei, Camellia semiserata, Liquidambar formosana, Castanopsis fissa, Mytilaria laosensis and Cinnamomum camphora seedlings were studied. Leaf N, P and K concentrations in Ormosia pinnata, Castanopsis fissa and Cinnamomum camphora were higher than those of other tree species, and leaf N, P and K contents in Camellia semiserata, Castanopsis fissa and Cinnamomum camphora were higher than those of other tree species. Mean leaf nutrient concentration in upper crown and lower crown of 9 tree species was N 16.67 and 17.09 g.kg-1, P 1.29 and 0.84 g.kg-1, K 11.77 and 6.65 g.kg-1, respectively. Mean leaf nutrient content in upper crown and lower crown of 9 tree species was N 0.871 and 1.069 g.m-2, P 0.065 and 0.052 g.m-2, K 0.608 and 0.426 g.m-2, respectively. Leaf P and K concentrations and contents in the upper crown were generally higher than those of the lower crown. From the view of nutrient contents, mixing Camellia semiserata with other tree species, mixing Castanopsis hystrix or Mytilaria laosensis with Elaeocarpus sylvestris, Liquidambar formosana, Ormosia pinnata, Castanopsis fissa, and Cinnamomum camphora, and mixing Michelia macclurei with Castanopsis fissa and Cinnamomum camphora were suitable.