Importance of cell division angle, position of cell proliferative area, and localization of AN3 in lateral organ morphology

Leaf meristem is a cell proliferative zone present in the lateral organ primordia, and it contributes to the expansion of lateral organ lamina. In this study, we investigated how the proliferative zone affects the final morphology of the lateral organs. We examined how cell proliferative zones differ in the primordia of polar-auxin transport inhibitor (PATI)-treated leaves and floral organs from normal foliage leaf primordia of Arabidopsis thaliana with focus on the spatial accumulation pattern of mRNA and protein of ANGUSTIFOLIA3 (AN3), a key element for leaf meristem positioning. As a result, we revealed that organ shape change by PATI treatment could not be attributed to changes in leaf-meristem positioning, size of the leaf meristem, or the expression pattern of AN3. Instead, it was attributed to altered cell division angles in the leaf meristem. In contrast, different shapes between sepals and petals compared with foliage leaves were observed to be correlated with both altered meristem position associated with altered AN3 expression patterns and different distributions of cell division angles. These results strongly indicate that lateral organ shapes are regulated via two aspects: position of meristem and cell division angles; the former is mainly governed by the AN3 expression pattern.

Integrated Transcriptome and Metabolome Analysis Reveals an Essential Role for Auxin in Hypocotyl Elongation during End-of-Day Far-Red Treatment of Cucurbita moschata (Duch. Ex Lam.)

Long, robust hypocotyls are important for facilitating greenhouse transplant production. The use of far-red light at the end of the day (end-of-day far-red, EOD-FR) is known to prompt hypocotyl elongation, but the mechanism of EOD-FR-mediated hypocotyl elongation in pumpkin remains unclear. Here, we found that hypocotyl length, parenchymal cell size in hypocotyls, and plant IAA levels were significantly greater in pumpkin after EOD-FR treatment. This effect was counteracted by the application of the polar auxin transport inhibitor 1-N-naphthylphthalamic acid. Integrated transcriptomic and metabolomic analysis of pumpkin hypocotyls revealed that the expression of auxin-related genes changed significantly after EOD-FR treatment, and the contents of the auxin biosynthetic precursors tryptophan and indole were also significantly higher. Our results show that auxin plays an essential role in EOD-FR-mediated hypocotyl elongation, shed light on the mechanisms of EOD-FR mediated hypocotyl elongation, and provide a theoretical basis for the use of EOD-FR in facility cultivation.

Auxin regulated metabolic changes underlying sepal retention and development after pollination in spinach

Background Pollination accelerate sepal development that enhances plant fitness by protecting seeds in female spinach. This response requires pollination signals that result in the remodeling within the sepal cells for retention and development, but the regulatory mechanism for this response is still unclear. To investigate the early pollination-induced metabolic changes in sepal, we utilize the high-throughput RNA-seq approach. Results Spinach variety ‘Cornel 9’ was used for differentially expressed gene analysis followed by experiments of auxin analog and auxin inhibitor treatments. We first compared the candidate transcripts expressed differentially at different time points (12H, 48H, and 96H) after pollination and detected significant difference in Trp-dependent auxin biosynthesis and auxin modulation and transduction process. Furthermore, several auxin regulatory pathways i.e. cell division, cell wall expansion, and biogenesis were activated from pollination to early developmental symptoms in sepals following pollination. To further confirm the role auxin genes play in the sepal development, auxin analog (2, 4-D; IAA) and auxin transport inhibitor (NPA) with different concentrations gradient were sprayed to the spinach unpollinated and pollinated flowers, respectively. NPA treatment resulted in auxin transport weakening that led to inhibition of sepal development at concentration 0.1 and 1 mM after pollination. 2, 4-D and IAA treatment to unpollinated flowers resulted in sepal development at lower concentration but wilting at higher concentration. Conclusion We hypothesized that sepal retention and development might have associated with auxin homeostasis that regulates the sepal size by modulating associated pathways. These findings advanced the understanding of this unusual phenomenon of sepal growth instead of abscission after pollination in spinach.

Crosstalk between auxin and gibberellin during stalk elongation in flowering Chinese cabbage

Plant growth and development are tightly regulated by phytohormones. However, little is known about the interaction between auxin and gibberellin acid (GA) during flower stalk elongation and how it is directly related to organ formation. Therefore, the effects of indole acetic acid (IAA) and GA3 treatments and their interaction on flower stalk elongation in flowering Chinese cabbage were investigated. The growth of flowering Chinese cabbage is regulated by IAA and GA3, and the opposite results were observed after treatments with uniconazole (GA synthesis inhibitor) and N-1-naphthylphthalamic acid (NPA) (auxin transport inhibitor). Anatomical analysis of the pith region in stalks revealed that IAA promoted expansion via signal transduction and transport pathways. GA3 regulated the elongation of flower stalks by controlling GA synthesis and partially controlling the IAA signaling pathway. GA3 also had a stronger effect on stalk elongation than IAA. The results of qRT-PCR and histological analysis revealed that GA3 and IAA induced the expansion of cell walls by activating the expression of genes encoding cell wall structural proteins such as Expansin (EXP). These findings provide new insights into the mechanism of stalk formation regulated by the combination of IAA and GA3.

Herbicide mixtures control glyphosate-resistant kochia (Bassia scoparia) in chemical fallow, but their longevity warrants careful stewardship

Glyphosate-resistant kochia [Bassia scoparia (L.) A.J. Scott], the first known glyphosate-resistant weed in western Canada, was confirmed initially in chemical fallow fields located in Warner County, Alberta in 2011. Further selection, lack of control, and rampant spread of this biotype contributed to its increased incidence, now present in about 50% of kochia populations sampled in Alberta. In 2014 and 2015, herbicide mixtures were evaluated based on control of glyphosate-resistant and susceptible kochia in chemical fallow fields near Lethbridge and Coalhurst, Alberta. The most consistent control (≥ 80% visual control in all environments with ≥ 80% biomass reduction in 2014) was observed with glyphosate + dicamba (450 + 580 g ae ha-1), glyphosate + dicamba/diflufenzopyr (450 + 150/50 g ai/ae ha-1), glyphosate + saflufenacil (450 + 50 g ai/ae ha-1), and glyphosate + carfentrazone + sulfentrazone (450 + 9 + 105 g ai/ae ha-1). Reduced efficacy was observed for several herbicide mixtures when they were applied to glyphosate-resistant compared with glyphosate-susceptible kochia accessions. Effective modes of action mixed with glyphosate include synthetic auxins (group 4), a combination of a synthetic auxin and an auxin transport inhibitor (group 19), or protoporphyrinogen oxidase inhibitors (group 14). In response to glyphosate-resistant kochia, many farmers in this region shifted their herbicide programs resulting in greater reliance on synthetic auxins; likely contributing to the recent discovery of auxinic herbicide-resistant kochia biotypes in Alberta in 2017. Careful herbicide stewardship is warranted to mitigate further selection of multiple herbicide-resistant kochia, suggesting an important role for integrated weed management.

Auxin Regulation and MdPIN Expression during Adventitious Root Initiation in Apple Cuttings

Adventitious root (AR) formation is critical for the efficient propagation of elite horticultural and forestry crops. Despite decades of research, the cellular processes and molecular mechanisms underlying AR induction in woody plants remains obscure. We examined the details of AR formation in the apple (Malus domestica) M.9 rootstock, the most widely used dwarf rootstock for intensive production, and investigated the role of polar auxin transport in post-embryonic organogenesis. AR formation begins with a series of founder cell divisions and elongation of interfascicular cambium adjacent to vascular tissues. This process was associated with a relatively high indole acetic acid (IAA) content and hydrolysis of starch grains. Exogenous auxin treatment promoted cell division, as well as the proliferation and reorganization of the endoplasmic reticulum and Golgi membrane. By contrast, treatment with the auxin transport inhibitor N-1-naphthylphthalamic acid (NPA) inhibited cell division in the basal region of the cutting and resulted in abnormal cell divisions during early AR formation. In addition, PIN-FORMED (PIN) transcripts were expressed differentially throughout the whole AR development process, with the up-regulation of MdPIN8 and MdPIN10 during induction, an up-regulation of MdPIN4, MdPIN5 and MdPIN8 during extension, and an up-regulation of all MdPINs during AR initiation. This research provides a deeper understanding of the cellular and molecular underpinnings of the AR process in woody plants.

Expression of a Rap2.12 like-1 ERF gene during adventitious rooting of chestnut and oak microshoots

Adventitious rooting of cuttings is a complex developmental process in forest species, with several exogenous and endogenous factors influencing the outcome of the process. In this study we applied an in vitro working system, comprising two lines of microshoots with the same genotype but at a different ontogenetic stages, in two different tree species (chestnut and oak). We analyzed the expression of a gene encoding an AP2/ERF transcription factor from group VII in the initial hours of the adventitious rooting induction, both in rooting competent and incompetent microshoots. The analysis revealed that expression of this gene is related to wounding, ontogenetic stage and auxin in a complex and species-specific manner. Putative induction of the gene by auxin was also analyzed in the presence of naphthyl-phthalamic acid (NPA), an auxin transport inhibitor. In situ expression analysis in chestnut relates the gene activity to cambial divisions and root primordia in rooting competent microshoots, as well as in the root apex. The putative role of the gene during adventitious roots formation is discussed.

Fine control of aerenchyma and lateral root development through AUX/IAA- and ARF-dependent auxin signaling

Lateral roots (LRs) are derived from a parental root and contribute to water and nutrient uptake from the soil. Auxin/indole-3-acetic acid protein (AUX/IAA; IAA) and auxin response factor (ARF)-mediated signaling are essential for LR formation. Lysigenous aerenchyma, a gas space created by cortical cell death, aids internal oxygen transport within plants. Rice (Oryza sativa) forms lysigenous aerenchyma constitutively under aerobic conditions and increases its formation under oxygen-deficient conditions; however, the molecular mechanisms regulating constitutive aerenchyma (CA) formation remain unclear. LR number is reduced by the dominant-negative effect of a mutated AUX/IAA protein in the iaa13 mutant. We found that CA formation is also reduced in iaa13. We have identified ARF19 as an interactor of IAA13 and identified a lateral organ boundary domain (LBD)-containing protein (LBD1-8) as a target of ARF19. IAA13, ARF19, and LBD1-8 were highly expressed in the cortex and LR primordia, suggesting that these genes function in the initiation of CA and LR formation. Restoration of LBD1-8 expression recovered aerenchyma formation and partly recovered LR formation in the iaa13 background, in which LBD1-8 expression was reduced. An auxin transport inhibitor suppressed CA and LR formation, and a natural auxin stimulated CA formation in the presence of the auxin transport inhibitor. Our findings suggest that CA and LR formation are both regulated through AUX/IAA- and ARF-dependent auxin signaling. The initiation of CA formation lagged that of LR formation, which indicates that the formation of CA and LR are regulated differently by auxin signaling during root development in rice.

Effects of auxin-transport-inhibitor and defoliation on wood formation in locally-heated Abies homolepis

ABSTRACTTo understand the precise process of wood formation, it is necessary to identify the factors that regulate cambial activity and development of cambial derivatives. Here, we investigated the combined effects of localized-heating and auxin on cambial reactivation and the formation of earlywood tracheids in seedlings of the evergreen coniferAbies homolepisin winter. Three treatments were applied, namely heating (artificial increase in temperature 20–22 °C), heating-plus-auxin transport inhibitor N-(1-naphthyl) phthalamic acid (NPA) and heating-plus-defoliation (removal of needles and buds), with an approximate control, for investigations of cambial activity by light microscopy. After one week of heating, cambial reactivation occurred in the heating, heating-plus-NPA and heating-plus-defoliation treatments. In untreated controls, cambial reactivation occurred later than in heated stems. Earlywood tracheids were formed after three and six weeks of heating in the heating and heating-plus-NPA treatments, respectively. No tracheids were formed after eight weeks of heating in heated-defoliated seedlings. Numbers of new tracheids were reduced in heated stems by NPA. Our results suggest that an increase in the temperature of the stem is one of the most important limiting factors in cambial reactivation, which is independent of needles and buds and of the polar transport of auxin from apical sources. However, after cambial reactivation, initiation and continuous formation of earlywood tracheids require basipetally transported auxin and other endogenous factors originating in mature needles and buds.