Roles for a soybean RAV-like orthologue in shoot regeneration and photoperiodicity inferred from transgenic plants.

The soybean gene Glyma10g34760 appears to encode a RAV2-like transcription factor orthologue (DQ147914; hereafter GmRAV) based on sequence similarity. The gene is a member of the ERF/AP2 transcription factor family that has been shown to be increased in transcript abundance by cytokinins (CKs). Transgenic GmRAV-overexpressing (-ox) tobacco plants exhibited increased CK signalling-related phenotypes including dwarfism, reduced apical dominance, extreme longevity, vigorous outgrowth of lateral buds, small and dark green leaves, reduced root growth, repressed flowering under both long- and short-day conditions, and altered sensitivity to daylength. In contrast, inhibition (-i) of GmRAV in soybean displayed the opposite phenotypic alterations which were consistent with defects in CK signalling. Phenotypes included earlier time of emergence; reduced numbers of branches, leaves, and flower buds; increased plant height; increased apical dominance; and earlier flowering and maturity. GmRAV-i soybean was less sensitive to cytokinin in hypocotyls and root growth inhibition assays. GmRAV-i soybean showed decreased frequency of adventious shoot formation in tissue culture in the presence of CKs, which might be attributed to the significantly decreased activities of CUC2, STM, and WUS involved in shoot meristem specification. GmRAV protein was localized in the nucleus in leaves. The GmRAV promoter-ß-glucuronidase (GUS) fusion was largely expressed in a meristematic region of the shoot apex, which was consistent with expressed sequence tag and microarray data. GmRAV was inferred to play a key role in CK and photoperiod signalling that subsequently regulated plant development.

A GmRAV ortholog is involved in photoperiod and sucrose control of flowering time in soybean.

Photoperiod and sucrose levels play a key role in the control of flowering. GmRAV reflected a diurnal rhythm with the highest expression at 4 h after the beginning of a dark period in soybean leaves, and was highly up-regulated under short-day (SD) conditions, despite of not following a diurnal pattern under long-day (LD) conditions. GmRAV-i (GmRAV-inhibition) transgenic soybean exhibited early flowering phenotype. Two of the FT Arabidopsis homologs, GmFT2a and GmFT5a, were highly expressed in the leaves of soybeans with inhibition (-i) of GmRAV under SD conditions. Moreover, the transcript levels of the two FT homologs in GmRAV-i soybeans were more sensitive to SD conditions than LD conditions compared to the WT plant. GmRAV-i soybeans and Arabidopsis rav mutants showed more sensitive hypocotyl elongation responses when compared with wild-type seedlings, and GmRAV-ox overevpressed in tobacco revealed no sensitive changes in hypocotyl length. These indicated that GmRAV was a novel negative regulator of SD-mediated flowering and hypocotyl elongation. Although sucrose has been suggested to promote flowering induction in many plant species, high concentration of sucrose (4% [w/v]) applied into media defer flowering time in Arabidopsis wild-type and rav mutant. This delayed flowering stage might be caused by reduction of LEAFY expression. Furthermore, Arabidopsis rav mutants and GmRAV-i soybean plants were less sensitive to sucrose by the inhibition assays of hypocotyls and roots growth. In contrast, transgenic GmRAV overexpressing (-ox) tobacco plants displayed more sensitivity to sucrose. In conclusion, GmRAV was inferred to have a fundamental function in photoperiod, darkness, and sucrose signaling responses to regulate plant development and flowering induction.