Background: Actin depolymerizing factors (ADFs) are small actin-binding proteins. Many higher-plant ADFs has been known to involve in plant growth, development and pathogen defense. However, in rice the temporal and spatial expression of OsADF gene family and their relationship with abiotic stresses tolerance is still unknown. Results: Here we reported the first comprehensive gene expression profile analysis of OsADF gene family. The OsADF genes showed distinct and overlapping gene expression patterns at different growth stages, tissues and abiotic stresses. We also demonstrated that both OsADF1 and OsADF3 proteins were localized in the nucleus. OsADF1 and OsADF3 were preferentially expressed in vascular tissues. Under ABA or abiotic stress treatments, OsADF3::GUS activity was enhanced in lateral roots and root tips. Ectopically overexpressed OsADF3 conferred the mannitol- and drought-stress tolerance of transgenic Arabidopsis seedlings by increasing germination rate, primary root length and survival. Several drought-tolerance responsive genes (RD22, ABF4, DREB2A, RD29A, PIP1; 4 and PIP2; 6) were upregulated in transgenic Arabidopsis under drought stress. Conclusions: These results suggested that OsADF gene family may participate in plant abiotic stresses response or tolerance and would facilitate functional validation of other OsADF genes.

Shoot regeneration frequency in rice callus is still low and highly diverse among rice cultivars. This study aimed to investigate the association of plant hormone signaling and sucrose uptake and metabolism in rice during callus induction and early shoot organogenesis. The immatured seeds of two rice cultivars, Ai-Nan-Tsao 39 (ANT39) and Tainan 11 (TN11) are used in this study. Callus formation is earlier, callus fresh weight is higher, but water content is significant lower in ANT39 than in TN11 while their explants are inoculated on callus induction medium (CIM). Besides, the regeneration frequency is prominently higher in ANT39 (~80%) compared to TN11 callus (0%). Levels of glucose, sucrose, and starch are all significant higher in ANT39 than in TN11 either at callus induction or early shoot organogenesis stage. Moreover, high expression levels of Cell wall-bound invertase 1, Sucrose transporter 1 (OsSUT1) and OsSUT2 are detected in ANT39 at the fourth-day in CIM but it cannot be detected in TN11 until the tenth-day. It suggested that ANT39 has higher callus growth rate and shoot regeneration ability may cause from higher activity of sucrose uptake and metabolism. As well, the expression levels of ORYZA SATIVA RESPONSE REGULATOR 1 (ORR1), PIN-formed 1 and Late embryogenesis-abundant 1, representing endogenous cytokinin, auxin and ABA signals, respectively, were also up-regulated in highly regenerable callus, ANT39, but only ORR1 was greatly enhanced in TN11 at the tenth-day in CIM. Thus, phytohormone signals may affect sucrose metabolism to trigger callus initiation and further de novo shoot regeneration in rice culture.

This study aimed to clarify the possible mechanism of endogenous phytohormone signaling and carbohydrate metabolism during shoot organogenesis induced by osmotic stress in rice (Oryza sativa L. cv. Tainung 71) callus. Non-regenerable calli derived from Tainung 71 immature embryos were inoculated on Murashige and Skoog medium containing 10 μM 2,4-D. They turned to highly regenerable calli (HRC) (regeneration frequency more than 75 %) with lower calli fresh weight and water content when 0.6 M sorbitol was supplemented into the medium. The regeneration frequency was prominently decreased to 25 % while an auxin transport inhibitor, 2,3,5-triiodobenzoic acid (TIBA), was added into the sorbitol-treated medium. It suggested that endogenous auxin signal may be involved in the induction of HRC under osmotic stress treatment. As well, HRC showed high levels of glucose, sucrose, and starch and high expression of cell wall-bound invertase 1, sucrose transporter 1 (OsSUT1), OsSUT2, PIN-formed 1, and late embryogenesis abundant 1 (OsLEA1) genes. Their expressions are all dramatic inhibited except OsLEA1 under TIBA treatment. It suggests a key role of auxin may be linked to the effect of shoot regeneration under osmotic stress treatment. Therefore, we present a putative hypothesis for regenerable calli induction by osmotic stress treatment in rice. Osmotic stress may regulate endogenous levels of auxin interacting with abscisic acid, then affect carbohydrate metabolism to trigger callus initiation and further shoot regeneration in rice.