Chemical induction of hypocotyl rooting reveals extensive conservation of auxin signalling controlling lateral and adventitious root formation.

Upon exposure to light, etiolated Arabidopsis seedlings form adventitious roots (AR) along the hypocotyl. While processes underlying lateral root formation are studied intensively, comparatively little is known about the molecular processes involved in the initiation of hypocotyl AR. AR and LR formation were studied using a small molecule named Hypocotyl Specific Adventitious Root INducer (HYSPARIN) that strongly induces AR but not LR formation. HYSPARIN does not trigger rapid DR5-reporter activation, DII-Venus degradation or Ca2+ signalling. Transcriptome analysis, auxin signalling reporter lines and mutants show that HYSPARIN AR induction involves nuclear TIR1/AFB and plasma membrane TMK auxin signalling, as well as multiple downstream LR development genes (SHY2/IAA3, PUCHI, MAKR4 and GATA23). Comparison of the AR and LR induction transcriptome identified SAURs, AGC kinases and OFP transcription factors as specifically upregulated by HYSPARIN. Members of the SAUR19 subfamily, OFP4 and AGC2 suppress HYS-induced AR formation. While SAUR19 and OFP subfamily members also mildly modulate LR formation, AGC2 regulates only AR induction. Analysis of HYSPARIN-induced AR formation uncovers an evolutionary conservation of auxin signalling controlling LR and AR induction in Arabidopsis seedlings and identifies SAUR19, OFP4 and AGC2 kinase as novel regulators of AR formation.

Genetic Dissection of Light-Regulated Adventitious Root Induction in Arabidopsis thaliana Hypocotyls.

Photomorphogenic responses of etiolated seedlings include the inhibition of hypocotyl elongation and opening of the apical hook. In addition, dark-grown seedlings respond to light by the formation of adventitious roots (AR) on the hypocotyl. How light signaling controls adventitious rooting is less well understood. Hereto, we analyzed adventitious rooting under different light conditions in wild type and photomorphogenesis mutants in Arabidopsis thaliana. Etiolation was not essential for AR formation but raised the competence to form AR under white and blue light. The blue light receptors CRY1 and PHOT1/PHOT2 are key elements contributing to the induction of AR formation in response to light. Furthermore, etiolation-controlled competence for AR formation depended on the COP9 signalosome, E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC (COP1), the COP1 interacting SUPPRESSOR OF PHYA-105 (SPA) kinase family members (SPA1,2 and 3) and Phytochrome-Interacting Factors (PIF). In contrast, ELONGATED HYPOCOTYL5 (HY5), suppressed AR formation. These findings provide a genetic framework that explains the high and low AR competence of Arabidopsis thaliana hypocotyls that were treated with dark, and light, respectively. We propose that light-induced auxin signal dissipation generates a transient auxin maximum that explains AR induction by a dark to light switch.

Arabidopsis Hypocotyl Adventitious Root Formation Is Suppressed by ABA Signaling.

Roots are composed of different root types and, in the dicotyledonous Arabidopsis, typically consist of a primary root that branches into lateral roots. Adventitious roots emerge from non-root tissue and are formed upon wounding or other types of abiotic stress. Here, we investigated adventitious root (AR) formation in Arabidopsis hypocotyls under conditions of altered abscisic acid (ABA) signaling. Exogenously applied ABA suppressed AR formation at 0.25 µM or higher doses. AR formation was less sensitive to the synthetic ABA analog pyrabactin (PB). However, PB was a more potent inhibitor at concentrations above 1 µM, suggesting that it was more selective in triggering a root inhibition response. Analysis of a series of phosphonamide and phosphonate pyrabactin analogs suggested that adventitious root formation and lateral root branching are differentially regulated by ABA signaling. ABA biosynthesis and signaling mutants affirmed a general inhibitory role of ABA and point to PYL1 and PYL2 as candidate ABA receptors that regulate AR inhibition.

Rhizobium symbiosis modulates the accumulation of arsenic in Medicago truncatula via nitrogen and NRT3.1-like genes regulated by ABA and linalool.

Arsenic (As) contamination is a worldwide problem and threatens human health. Here, we found that Rhizobium symbiosis can improve the tolerance to arsenate [As(V)], and a wild type R. meliloti Rm5038 symbiosis can significantly decrease the accumulation of As in Medicago truncatula shoots. The As content in plants could be decreased by nitrogen and the mutation of nitrate transporter NRT3.1. The expression of M. truncatula NRT3.1-like gene NRT3.1L1 could reverse the As(V)-tolerance phenotype of the Arabidopsis nrt3.1 mutant. Rm5038 symbiosis significantly increased the level of nitrogen in the shoot and reduced the expression of NRT3.1Ls in plants afflicted by As(V). The genetic analyses of aba2-1, pyr1/pyl1/2/4/5/8, and abi1-2/abi2-2/hab1-1/pp2ca-1 mutants revealed that abscisic acid (ABA) signaling regulates the tolerance of plants to As(V). ABA and linalool could promote the expression of NRT3.1Ls, however, their root biosynthesis was inhibited by ammonium, the first form of nitrogen fixed by Rhizobium symbiosis. Moreover, ABA and linalool may also control As and nitrate accumulation in Rhizobium symbionts via signaling pathways other than ammonia and NRT3.1Ls. Thus, Rhizobium symbiosis modulates the accumulation of As in plants via nitrogen and NRT3.1Ls regulated by ABA and linalool, which provides novel approaches to reduce As accumulation in legume crops.