Nitrate reductase (NR) is an enzymatic source of nitric oxide (NO) in plants, and it needs Mo for the Mo-cofactor to be activated. Because NR-deficient mutants are not always available in some species, a cheap and simple pharmacological application of tungstate, which substitutes for Mo in the Mo-cofactor as a competitive antagonist, is widely used as a NR inhibitor in plant NO research. However, evidence indicates that tungstate not only inactivates NR but also inhibits other molybdate-dependent enzymes in plants. In addition, a number of investigations have shown that tungstate also inhibits root growth, affects cortical microtubule formation, and induces programmed cell death (PCD) in plants, just like other heavy metals do. Therefore, tungstate has been shown to exert many other effects that are not connected with the inhibition of NR activity. The origin and mechanism of using tungstate as a NR inhibitor in plants is reviewed here and the progress regarding tungstate toxicity to plants and the possible problems involved in using tungstate as a NR inhibitor in plant NO research are analysed. In summary, the use of tungstate as a NR inhibitor in plant NO research must be treated with caution, keeping in mind that it is not completely specific. It is necessary to search for more NR-deficient mutants and new, specific NR inhibitors. A combination of pharmacological and biochemical analysis with a genetic approach will be necessary in order to investigate the roles of NO in plants.

Iron (Fe) is essential for rice growth and humans consuming as their staple food but is often deficient because of insoluble Fe(III) in soil for rice growth and limited assimilation for human bodies, while cadmium (Cd) is non-essential and toxic for rice growth and humans if accumulating at high levels. Over-accumulated Cd can cause damage to human bodies. Selecting and breeding Fe-rich but Cd-free rice cultivars are ambitious, challenging and meaningful tasks for researchers. Although evidences show that the mechanisms of Fe/Cd uptake and accumulation in rice are common to some extent as a result of similar entry routes within rice, an increasing number of researchers have discovered distinct mechanisms between Fe/Cd uptake and accumulation in rice. This comprehensive review systematically elaborates and compares cellular mechanisms of Fe/Cd uptake and accumulation in rice, respectively. Mechanisms for maintaining Fe homeostasis and Cd detoxicification are also elucidated. Then, effects of different fertilizer management on Fe/Cd accumulation in rice are discussed. Finally, this review enumerates various approaches for reducing grain Cd accumulation and enhancing Fe content in rice. In summary, understanding of discrepant cellular mechanisms of Fe/Cd accumulation in rice provides guidance for cultivating Fe-fortified rice and has paved the way to develop rice that are tolerant to Cd stress, aiming at breeding Fe-rich but Cd-free rice.

Aeration and water management increasing rhizosphere oxygen amount significantly promote rice (Oryza sativa) growth and yield, but the effect of root aeration on cadmium (Cd) toxicity and accumulation in rice seedlings under hydroponic culture remains unclear. Results showed that aeration promoted rice seedling growth and alleviated Cd toxicity. Transverse section discovered that Cd accelerated root mature and senescence while aeration delayed the mature and senescence of roots. Non-invasive Micro-test Technology (NMT) showed that aeration increased net O2 and Cd2+ influxes on the surface of roots while decreased net Cd2+ influx in xylem. Perls blue staining showed that aeration and Cd treatments increased iron plaque formation on the surface of roots. Results of metal concentration analysis showed that besides increasing Cd retention in iron plaque, aeration also increasing Cd retention in the cell wall of rice roots. Cell wall component analysis showed that aeration not only increased pectin content but also decreased pectin methylesterification degree (PMD) by increasing pectin methylesterase (PME) activity. All of these results indicate that aeration not only delays root mature and senescence but also increases Cd retention in roots by enhancing iron plaque formation and regulating pectin synthesis in the roots of rice seedlings.