Bioenergy sorghum stem growth regulation: intercalary meristem localization, development, and gene regulatory network analysis.

Bioenergy sorghum is a highly productive drought tolerant C4 grass that accumulates 80% of its harvestable biomass in approximately 4 m length stems. Stem internode growth is regulated by development, shading, and hormones that modulate cell proliferation in intercalary meristems (IMs). In this study, sorghum stem IMs were localized above the pulvinus at the base of elongating internodes using magnetic resonance imaging, microscopy, and transcriptome analysis. A change in cell morphology/organization occurred at the junction between the pulvinus and internode where LATERAL ORGAN BOUNDARIES (SbLOB), a boundary layer gene, was expressed. Inactivation of an AGCVIII kinase in DDYM (dw2) resulted in decreased SbLOB expression, disrupted IM localization, and reduced internode cell proliferation. Transcriptome analysis identified approximately 1000 genes involved in cell proliferation, hormone signaling, and other functions selectively upregulated in the IM compared with a non-meristematic stem tissue. This cohort of genes is expressed in apical dome stem tissues before localization of the IM at the base of elongating internodes. Gene regulatory network analysis identified connections between genes involved in hormone signaling and cell proliferation. The results indicate that gibberellic acid induces accumulation of growth regulatory factors (GRFs) known to interact with ANGUSTIFOLIA (SbAN3), a master regulator of cell proliferation. GRF:AN3 was predicted to induce SbARF3/ETT expression and regulate SbAN3 expression in an auxin-dependent manner. GRFs and ARFs regulate genes involved in cytokinin and brassinosteroid signaling and cell proliferation. The results provide a molecular framework for understanding how hormone signaling regulates the expression of genes involved in cell proliferation in the stem IM.

Histopathology of the Shoot Apex of Sugarcane Colonized by Leifsonia xyli subsp. xyli.

Colonization of the xylem of sugarcane by Leifsonia xyli subsp. xyli results in the occlusion of the vessels by a gum-like compound and compromises the elongation of the stalk leading to stunted plants. However, no study has been performed in the apical tissue where the elongation of the stalks initiates at the intercalary meristem (IM). Microscopic and histochemical analyses were performed in plants with lower and higher bacterial titers and revealed that in both cases L. xyli subsp. xyli is present in this tissue and colonizes the forming xylem vessels in a similar way as observed in developed internodes. In both cases, it was observed adhering to the secondary walls, but only in plants with higher titers were a mild degradation of the walls and a granular material filling the vessels observed. The mixed composition of lipids, proteins, and pectin indicates that the filling is not a bacterial extracellular polymeric substance. Plants with higher bacterial populations also presented lower starch content in the ground parenchyma at the node elements, possibly resulting from the reported downregulation of photosynthesis and increased accumulation of phenolics. Their second and third IMs presented fewer cells and reduced expression of genes related to the cell cycle and to the synthesis of ABA in the apical tissue. These results indicate that increased L. xyli subsp. xyli colonization affects the development of the IM, which ultimately would reduce the length of the internodes, resulting in the main symptom of the disease.

Cellular and molecular characterizations of the irregular internode division zone formation of a slow-growing bamboo variant.

The key molecular mechanisms underlying the sectionalized growth within bamboo or other grass internodes remain largely unknown. Here, we genetically and morphologically compared the culm and rhizome internode division zones (DZs) of a slow-growing bamboo variant (sgv) having dwarf internodes, with those of the corresponding wild type (WT). Histological analysis discovers that the sgv has an irregular internode DZ. However, the shoot apical meristems in height, width, outside shape, cell number and cell width of the sgv and the WT were all similar. The DZ irregularities first appeared post apical meristem development, in 1-mm sgv rhizome internodes. Thus, the sgv is a DZ irregularity bamboo variant, which has been first reported in bamboo according to our investigation. Transcriptome sequencing analysis finds that a number of cell wall biogenesis and cell division-related genes are dramatically downregulated in the sgv DZ. Interestingly, both transcriptomic and brassinosteroid (BR) contents detection, as well as quantitative real-time PCR analyses show that these irregularities have resulted from the BR signaling pathway defects. Brassinosteroid defect might also cause the erect leaves and branches as well as the irregular epidermis of the sgv. These results suggest that BR signaling pathway plays critical roles in bamboo internode DZ and leaf development from a mutant perspective and also explain the upstream mechanisms causing the dwarf internode of the sgv bamboo.

Moving on up - controlling internode growth.

Plant reproductive success depends on making fertile flowers but also upon developing appropriate shoot internodes that optimally arrange and support the flowering shoot. Compared to floral morphogenesis, we understand little about the networks directing internode growth during flowering. However, new studies reveal that long-range signals, local factors, and age-dependent micoRNA-networks are all important to harmonize internode morphogenesis with shoot development. Some of the same players modulate symplastic transport to seasonally regulate internode growth in perennial species. Exploring possible hierarchical control amongst symplastic continuity, age, systemic signals and local regulators during internode morphogenesis will help elucidate the mechanisms coordinating axial growth with the wider plant body.

Maize brachytic2 (br2) suppresses the elongation of lower internodes for excessive auxin accumulation in the intercalary meristem region.

BACKGROUND: Short internodes contribute to plant dwarfism, which is exceedingly beneficial for crop production. However, the underlying mechanisms of internode elongation are complicated and have been not fully understood. RESULTS: Here, we report a maize dwarf mutant, dwarf2014 (d2014), which displays shortened lower internodes. Map-based cloning revealed that the d2014 gene is a novel br2 allele with a splicing variation, resulting in a higher expression of BR2-T02 instead of normal BR2-T01. Then, we found that the internode elongation in d2014/br2 exhibited a pattern of inhibition-normality-inhibition (transient for the ear-internode), correspondingly, at the 6-leaf, 12-leaf and 14-leaf stages. Indeed, BR2 encodes a P-glycoprotein1 (PGP1) protein that functions in auxin efflux, and our in situ hybridization assay showed that BR2 was mainly expressed in vascular bundles of the node and internode. Furthermore, significantly higher auxin concentration was detected in the stem apex of d2014 at the 6-leaf stage and strictly in the node region for the ear-internode at the 14-leaf stage. In such context, we propose that BR2/PGP1 transports auxin from node to internode through the vascular bundles, and excessive auxin accumulation in the node (immediately next to the intercalary meristem) region suppresses internode elongation of d2014. CONCLUSIONS: These findings suggest that low auxin levels mediated by BR2/PGP1 in the intercalary meristem region are crucial for internode elongation.

Characterization of the developmental dynamics of the elongation of a bamboo internode during the fast growth stage.

Previous studies on the fast growth of bamboo shoots mainly focused on the entire culm. No work about the fast elongation of a single internode, which is the basic unit for the fast growth of bamboo shoots, has been reported so far according to our knowledge. In this study, we have systematically investigated the regulating mechanisms underlying the fast growth of a single bamboo internode of Bambusa multiplex (Lour.) Raeusch. ex Schult. We discovered that the growth of the internode displays a logistic pattern, and the two sections located in the bottom of the internode, one for cell division and, another for cell elongation, each with an ~1-cm length, comprise the effective zones for the internode growth. RNA-Seq analysis identified a number of genes potentially involved in regulating the fast growth of bamboo internode such as those that have positive roles in promoting cell growth or division, which were dramatically down-regulated in the internode at fast growth decreasing stage. Further analysis revealed that sugar plays an important role in promoting the fast growth of bamboo internodes through inhibition of BmSnf1. Mechanical stress is found to be involved in the triggering of the internode growth decrease through activation of the generation of reactive oxygen species by upregulating Calmodulins. These results provide systematic insight into the biological mechanisms underlying the fast growth of bamboo shoots based on the behavior of a single internode.

APETALA2 control of barley internode elongation.

Many plants dramatically elongate their stems during flowering, yet how this response is coordinated with the reproductive phase is unclear. We demonstrate that microRNA (miRNA) control of APETALA2 (AP2) is required for rapid, complete elongation of stem internodes in barley, especially of the final 'peduncle' internode directly underneath the inflorescence. Disrupted miR172 targeting of AP2 in the Zeo1.b barley mutant caused lower mitotic activity, delayed growth dynamics and premature lignification in the peduncle leading to fewer and shorter cells. Stage- and tissue-specific comparative transcriptomics between Zeo1.b and its parent cultivar showed reduced expression of proliferation-associated genes, ectopic expression of maturation-related genes and persistent, elevated expression of genes associated with jasmonate and stress responses. We further show that applying methyl jasmonate (MeJA) phenocopied the stem elongation of Zeo1.b, and that Zeo1.b itself was hypersensitive to inhibition by MeJA but less responsive to promotion by gibberellin. Taken together, we propose that miR172-mediated restriction of AP2 may modulate the jasmonate pathway to facilitate gibberellin-promoted stem growth during flowering.

Efficient regeneration system for rapid multiplication of clean planting material of Ensete ventricosum (Welw.) Cheesman.

Enset (Ensete ventricosum (Welw.) Cheesman) is an economically important staple food crop in Ethiopia, especially in the southern and southwestern regions. It is called "false banana" due to its resemblance to banana, but inability to produce any edible fruit. The crop is clonally propagated using field-grown suckers. This study reports the development of a robust regeneration technique to propagate large numbers of plantlets using corm discs containing intercalary meristematic tissues. Hundreds of shoot buds were induced from corm discs of enset cultivar 'Bedadeti' cultured on Murashige and Skoog (MS) medium supplemented with 1.5 mg L-1 2,4-dichlorophenoxyacetic acid, 0.216 mg L-1 zeatin, and 2 g L-1 activated charcoal. The shoot buds were regenerated into complete plantlets when transferred onto MS medium supplemented with 1 mg L-1 6-benzylaminopurine and 2 g L-1 activated charcoal. More than 100 plantlets were generated in 4 mo from corm discs isolated from a single in vitro mother plantlet. Well-rooted plantlets were acclimatized in soil with 100% success, and did not show any apparent phenotypic abnormalities under glasshouse conditions. This efficient regeneration system could be very useful for the rapid multiplication of clean pathogen-free planting material.

The ontogenetic bases for variation in ovary position in Melastomataceae.

PREMISE OF THE STUDY: Although the ovary position is considered a stable character in angiosperms, Melastomataceae species have perigynous flowers in which the ovary varies from superior to inferior. Thus, we investigated the ontogenetic process involved in variation of the ovary position in Melastomataceae. We focused on histogenesis of the floral apex in search of developmental patterns for each type of ovary position. METHODS: Six species in which the ovary varies from superior to inferior were chosen: Henriettea saldanhae, Leandra melastomoides, Miconia dodecandra, Microlicia euphorbioides, Rhynchanthera grandiflora, and Tibouchina clinopodifolia. Buds and flowers were processed for surface and histological examinations. KEY RESULTS: The floral apex changes from convex to concave, resulting in a perigynous hypanthium. Cell divisions in the margins of the floral apex form an annular intercalary meristem that elevates the base of the primordia of almost all whorls. The joint growth of the carpel base with the gynoecial hypanthium originates semi-inferior ovaries in Leandra melastomoides, Miconia dodecandra, and Tibouchina clinopodifolia and inferior ovaries in Henriettea saldanhae. In Microlicia euphorbioides and Rhynchanthera grandiflora, the carpels are not affected by this hypanthial growth; flowers have a superior ovary. CONCLUSIONS: Changes in ovary position of Melastomataceae are due to intercalary meristematic activity, which is one of the main mechanisms for the origin of morphological innovations among plants. Our data illustrate the importance of the intercalary meristems in floral development, and we discuss the implications of this ontogenetic model for understanding the evolution of ovary position in Melastomataceae.