Phloem loading is the first step in sucrose transport from source leaves to sink organs. The phloem loading strategy in rice remains unclear. To determine the potential phloem loading mechanism in rice, yeast invertase (INV) was overexpressed specifically in the cell wall by 35S promoter to block sugar transmembrane loading in rice. The transgenic lines exhibited obvious phloem loading suppression characteristics accompanied by the accumulation of sucrose and starch, restricted vegetative growth and decreased grain yields. The decreased sucrose exudation rate with p-chloromercuribenzenesulfonic acid (PCMBS) treatment also indicated that rice actively transported sucrose into phloem. Moreover, the expression level of OsSUT1 was much higher than that of other plasma membrane localized OsSUTs in the source leaf. Cross sections of the GUS transgenic plant showed that the signals of OsSUT1 and OsSUT5 occurred in the phloem companion cells. The ossut1 and ossut4 mutants presented a decrease of grain yield, implying important roles of OsSUTs in phloem loading. Based on these results, we conclude that rice uses the apoplastic loading as a major phloem loading strategy.

Iron (Fe) is an essential mineral element required for plant growth, and when soil availability of Fe is low, plants show symptoms of severe deficiency. Under conditions of Fe deficiency, plants alter several processes to acquire Fe from soil. In this study, we used rice cultivars H 9405 with high Fe accumulation in seeds and Yang 6 with low Fe accumulation in seeds to study their physiological responses to different conditions of Fe availability. In both shoots and roots, the responses of ROS enzymes, leaf and root ultrastructure and photosynthetic system to iron deficiency in Yang 6 were much sensitive than those in H 9405. For the distribution of iron, the iron content was much higher in roots of Yang 6, in contrast to higher shoot content in H 9405. Differential responses were shown with the Fe content in roots and shoots, which were the opposite in the two varieties; thus, we proposed the existence of long-distance signals. Then split root and shoot removal experiments were used to demonstrate that a long-distance signal was involved in the iron-deficient rice plant, and the signal strength was highly correlated with the functional leaves.

Sucrose signaling pathways were rapidly induced in response to early iron deficiency in rice plants, and the change of sucrose contents in plants was essential for the activation of iron deficiency responses. Sucrose is the main product of photosynthesis in plants, and it functions not only as an energy metabolite but also a signal molecule. However, a few studies have examined the involvement of sucrose in mediating iron deficiency responses in rice. In this study, we found that the decrease in photosynthesis and total chlorophyll concentration (SPAD values) in leaves occurred at a very early stage under iron deficiency. In addition, the sucrose was increased in leaves but decreased in roots of rice plants under iron deficiency, and also the sucrose transporter (SUT) encoded genes' expression levels in leaves were all inhibited, including OsSUT1, OsSUT2, OsSUT3, OsSUT4, and OsSUT5. The carbohydrate distribution was changed under iron deficiency and sucrose might be involved in the iron deficiency responses of rice plants. Furthermore, exogenous application of sucrose or dark treatment experiments were used to test the hypothesis; we found that the increased endogenous sucrose would cause the repression of iron acquisition-related genes in roots, while further stimulated iron transport-related genes in leaves. Compared to the exogenous application of sucrose, the dark treatment had the opposite effects. All the above results highlighted the important role of sucrose in regulating the responses of rice plants to iron deficiency.