Microarray analysis of vegetative phase change in maize

Vegetative phase change is the developmental transition from the juvenile phase to the adult phase in which a plant becomes competent for sexual reproduction. The gain of ability to flower is often accompanied by changes in patterns of differentiation in newly forming vegetative organs. In maize, juvenile leaves differ from adult leaves in morphology, anatomy and cell wall composition. Whereas the normal sequence of juvenile followed by adult is repeated with every sexual generation, this sequence can be altered in maize by the isolation and culture of the shoot apex from an adult phase plant: an 'adult' meristem so treated reverts to forming juvenile vegetative organs. To begin to unravel the as-yet poorly understood molecular mechanisms underlying phase change in maize, we compared gene expression in two juvenile sample types, leaf 4 and culture-derived leaves 3 or 4, with an adult sample type (leaf 9) using cDNA microarrays. All samples were leaf primordia at plastochron 6. A gene was scored as 'phase induced' if it was up- or downregulated in both juvenile sample types, compared with the adult sample type, with at least a twofold change in gene expression at a P-value of < or =0.005. Some 221 expressed sequence tags (ESTs) were upregulated in juveniles, and 28 ESTs were upregulated in adults. The largest class of juvenile-induced genes was comprised of those involved in photosynthesis, suggesting that maize plants are primed for energy production early in vegetative growth by the developmental induction of photosynthetic genes.     

The viviparous8 mutation delays vegetative phase change and accelerates the rate of seedling growth in maize

Post-embryonic shoot development in plants can be divided into a juvenile vegetative, an adult vegetative, and a reproductive phase, which are expressed in different domains on the shoot axis. The number and position of the phytomers in each phase are determined by the time at which a plant begins and ceases making phytomers of a particular phase and the rate at which phytomers are made during that phase. The viviparous8 (vp8) mutation of maize increases the number of juvenile vegetative phytomers and decreases the number of adult vegetative phytomers by affecting both of these processes. vp8 increases the number of juvenile vegetative phytomers by increasing the rate of leaf initiation early in shoot development and delaying the juvenile-to-adult transition (vegetative maturation). It reduces the number of adult phytomers because the delay in vegetative maturation is not matched by a corresponding delay in flowering time; vp8 plants produce a tassel at the same time as wild-type plants. Thus, Vp8 normally controls the production of a factor that functions both to repress the rate of growth early in shoot development and to promote vegetative maturation, but which has no major role in floral induction. vp8 dramatically enhances the phenotypes of the dwarf and Teopod mutants and requires a functional Glossy15 gene to prolong the expression of juvenile epidermal traits. Evidence suggesting that vp8 does not affect phase change by reducing the level of abscisic acid is discussed.     

miRNA control of vegetative phase change in trees

After germination, plants enter juvenile vegetative phase and then transition to an adult vegetative phase before producing reproductive structures. The character and timing of the juvenile-to-adult transition vary widely between species. In annual plants, this transition occurs soon after germination and usually involves relatively minor morphological changes, whereas in trees and other perennial woody plants it occurs after months or years and can involve major changes in shoot architecture. Whether this transition is controlled by the same mechanism in annual and perennial plants is unknown. In the annual forb Arabidopsis thaliana and in maize (Zea mays), vegetative phase change is controlled by the sequential activity of microRNAs miR156 and miR172. miR156 is highly abundant in seedlings and decreases during the juvenile-to-adult transition, while miR172 has an opposite expression pattern. We observed similar changes in the expression of these genes in woody species with highly differentiated, well-characterized juvenile and adult phases (Acacia confusa, Acacia colei, Eucalyptus globulus, Hedera helix, Quercus acutissima), as well as in the tree Populus x canadensis, where vegetative phase change is marked by relatively minor changes in leaf morphology and internode length. Overexpression of miR156 in transgenic P. x canadensis reduced the expression of miR156-targeted SPL genes and miR172, and it drastically prolonged the juvenile phase. Our results indicate that miR156 is an evolutionarily conserved regulator of vegetative phase change in both annual herbaceous plants and perennial trees.     

Gibberellins in apical shoot meristems of flowering and vegetative sugarcane

Gibberellins A₁, A₃, iso-A₃, A₄, A₁₉, A₂₀, and A₃₆ were identified by gas chromatography-selected ion monitoring in apices of sugarcane (Saccharum spp. hybrids). Flowering apices (i.e., 2–4 cm panicle) contained 8–9 times more (estimated by bioassay) endogenous gibberellins A and iso-GA₃ (ratio of 1:6:8, respectively; in total 51 ng g^–1 fresh weight) than vegetative apices (6.4 ng g^–1 fresh weight). Vegetative apices contained small but significant levels of GA₁₉, which could not be detected in flowering apices; vegetative apices also contained approximately four times more of a GA₃₆-like substance than flowering apices. Since the two apex types developed under the same photoperiod, the increased levels of GA and iso-GA₃ and the reduced levels of GA₁₉ and GA₃₆-like substances are correlated with the flowering state rather than with photoperiod or photoperiod changes per se. Since there were relatively high levels of C₁₉ GAs along with low levels of C₂₀ GAs in flowering apices, and since the converse is true in vegetative apices, metabolism of C₂₀ to C₁₉ GAs may be enhanced in flowering apices.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

The early phase change gene in maize

Recessive mutations of the early phase change (epc) gene in maize affect several aspects of plant development. These mutations were identified initially because of their striking effect on vegetative phase change. In certain genetic backgrounds, epc mutations reduce the duration of the juvenile vegetative phase of development and cause early flowering, but they have little or no effect on the number of adult leaves. Except for a transient delay in leaf production during germination, mutant plants initiate leaves at a normal rate both during and after embryogenesis. Thus, the early flowering phenotype of epc mutations is explained completely by their effect on the expression of the juvenile phase. The observation that epc mutations block the rejuvenation of leaf primordia in excised shoot apices supports the conclusion that epc is required for the expression of juvenile traits. This phenotype suggests that epc functions normally to promote the expression of the juvenile phase of shoot development and to suppress the expression of the adult phase and that floral induction is initiated by the transition to the adult phase. epc mutations are epistatic to the gibberellin-deficient mutation dwarf1 and interact additively with the dominant gain-of-function mutations Teopod1, Teopod2, and Teopod3. Genetic backgrounds that enhance the mutant phenotype of epc demonstrate that, in addition to its role in phase change, epc is required for the maintenance of the shoot apical meristem, leaf initiation, and root initiation.     

Gibberellins and heterosis in maize : I. Endogenous gibberellin-like substances

Under controlled environment and/or field conditions, vegetative growth (height, internode length, leaf area, shoot dry weight, grain yield) was greater in an F₁ maize hybrid than in either parental inbred. Endogenous gibberellin (GA)-like substances in apical meristem cylinders were also higher in the hybrid than in either inbred, both on a per plant and per gram dry weight basis. There were no apparent qualitative differences in GA-like substances, however. Levels of GA-like substances in all genotypes were highest prior to tassel initiation. Chromatographic comparisons of the GA-like substances and authentic standards of GA native to maize on gradient-eluted SiO₂ partition and reverse-phase C₁₈ high-pressure liquid chromatography columns are described. No consistent differences in abscisic acid levels of the three genotypes were observed. This correlation of heterosis for endogenous GA-like substances with heterosis for growth suggests that amounts of endogenous GA may be related to hybrid vigor in maize.     

Toward positional cloning of Vgt1, a QTL controlling the transition from the vegetative to the reproductive phase in maize

Vgt1 (Vegetative to generative transition 1) is a quantitative trait locus (QTL) for flowering time in maize (Zea mays L.). Vgt1 was initially mapped in a ca. 5-cM interval on chromosome bin 8.05, using a set of near-isogenic lines (NILs) in the genetic background of the late dent line N28, with the earliness allele introgressed from the early variety Gaspé Flint. A new large mapping population was produced by crossing N28 and one early NIL with a ca. 6-cM long Gaspé Flint introgression at the Vgt1 region. Using PCR-based assays at markers flanking Vgt1, 69 segmental NILs homozygous for independent crossovers near the QTL were developed. When the NILs were tested in replicated field trials for days to pollen shed (DPS) and plant node number (ND), the QTL followed a Mendelian segregation. Using bulk segregant analysis and AFLP profiling, 17 AFLP markers linked to the QTL region were identified. Statistical analysis indicated a substantial coincidence of the effects of Vgt1 on both DPS and ND. Vgt1 was mapped at ca. 0.3 cM from an AFLP marker. As compared to DPS, the higher heritability of ND allowed for a more accurate assessment of the effects of Vgt1. The feasibility of the positional cloning of Vgt1 is discussed.     

vanishing tassel2 encodes a grass-specific tryptophan aminotransferase required for vegetative and reproductive development in maize

Auxin plays a fundamental role in organogenesis in plants. Multiple pathways for auxin biosynthesis have been proposed, but none of the predicted pathways are completely understood. Here, we report the positional cloning and characterization of the vanishing tassel2 (vt2) gene of maize (Zea mays). Phylogenetic analyses indicate that vt2 is a co-ortholog of TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS1 (TAA1), which converts Trp to indole-3-pyruvic acid in one of four hypothesized Trp-dependent auxin biosynthesis pathways. Unlike single mutations in TAA1, which cause subtle morphological phenotypes in Arabidopsis thaliana, vt2 mutants have dramatic effects on vegetative and reproductive development. vt2 mutants share many similarities with sparse inflorescence1 (spi1) mutants in maize. spi1 is proposed to encode an enzyme in the tryptamine pathway for Trp-dependent auxin biosynthesis, although this biochemical activity has recently been questioned. Surprisingly, spi1 vt2 double mutants had only a slightly more severe phenotype than vt2 single mutants. Furthermore, both spi1 and vt2 single mutants exhibited a reduction in free auxin levels, but the spi1 vt2 double mutants did not have a further reduction compared with vt2 single mutants. Therefore, both spi1 and vt2 function in auxin biosynthesis in maize, possibly in the same pathway rather than independently as previously proposed.     

Gibberellins and bud break,vegetative shoot growth and flowering in Metrosideros collina cv. Tahiti

Applications of the growth promotive gibberellins (GAs) GA₄ and 2,2-dimethyl GA₄, and of C-16,17 endo-dihydro GA₅, which is known to promote flowering while inhibiting stem growth in the long-day grass Lolium temulentum, were made to micropropagated plants of Metrosideros collina cv. Tahiti, a highly ornamental cultivar with an intermittent flowering pattern. Gibberellin A₄ and 2,2-dimethyl GA₄ stimulated vegetative growth both in elongating shoots, and internodes of shoots developing from buds that were quiescent at the time of GA application. Abscission of the apices of expanding shoots, a feature of mature Metrosideros plants, was inhibited by these GAs, the rejuvenation of micropropagated plantlets being enhanced. However, C-16,17 endo-dihydro GA₅ differed from GA₄ and 2,2-dimethyl GA₄ by having no promotive effects on vegetative growth, and no inhibition of apical abscission. Notwithstanding this contrasting effect on vegetative growth, high doses of GA₄ or C-16,17 endo-dihydro GA₅ similarly reduced flowering on shoots to which either GA was applied. Reduced flowering in response to applied GAs is common in many woody angiosperms, and in this instance was probably the combined result of abortion of developing floral structures in quiescent buds, and a preferential inhibition of bud break for floral buds relative to vegetative buds, particularly by GA₄. Finally, both C-16,17 endo-dihydro GA₅ and GA₄ strongly inhibited bud break in this woody angiosperm, although GA₄ could initially stimulate bud break when applied to vegetative buds close to the expansion stage. The above findings, in toto, highlight the sensitivity of Metrosideros to both classes of GA in a variety of growth and development processes.     

The nuclear localization signal is required for the function of squamosa promoter binding protein-like gene 9 to promote vegetative phase change in Arabidopsis

Plant Molecular Biology - A mutation in the nuclear localization signal of squamosa promoter binding like-protein 9 (SPL9) delays vegetative phase change by disrupting its nuclear localization. The...     

Sunlight-induced anthocyanin pigmentation in maize vegetative tissues

Although, in maize, sunlight-regulated anthocyanin formation in vegetative tissues is observed only in the cultivars harbouring homozygous recessive pl loci, the identity of the photoreceptor mediating this process is not yet fully established. In this study the nature of photoreceptor(s) mediating this response was examined using an Indian hybrid maize cultivar (Kanchan-521). The etiolated maize seedlings of this cultivar on exposure to sunlight formed anthocyanin in all vegetative organs. Sunlight elicited photoinduction of anthocyanin with a slow increase between 4-16 h after the sunlight exposure, followed by a rapid increase between 16-24 h. The photoinduction of anthocyanin was primarily mediated by the UV-B component of sunlight and could be elicited by exposure to an artificial UV-B light source. The sunlight-mediated induction of anthocyanin was reduced if the sunlight exposure was terminated with a far-red pulse before transfer to darkness, indicating a coaction of phytochrome in this photoresponse. Exposure to sunlight also stimulated phenylalanine ammonia lyase (PAL) activity in all organs with two temporally separated peaks. The first peak of PAL between 4-12 h was induced by phytochrome, and the second peak of PAL between 12-24 h was induced by UV-B light. These results indicate that the photoinduction of anthocyanin in maize is mediated by a coaction of UV-B light and phytochrome.     

Differential proteomic analysis during the vegetative phase change and the floral transition in Malus domestica

In order to identify the proteomic changes of apple (Malus domestica Borkh.) during the vegetative phase change and the floral transition, leaf protein of juvenile, adult vegetative and reproductive phase in a seedling ('Jonathan' × 'Golden Delicious') was extracted and analyzed by 2-D electrophoresis and Matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Seventy two gel spots with significant expression differences between ontogenetic phases were obtained. Five protein spots were only detected in leaves of juvenile phase and 11 were not; 17 spots were found exclusively in adult vegetative leaves; and only one spot solely appeared in reproductive leaves while 12 did not. Twenty six of the differentially expressed proteins identified were involved in photosynthesis. Seven enzymes were related to respiration and carbohydrate metabolism. Fifteen other proteins also presented qualitative or quantitative differences among developmental phases. The spatial distribution of one differentially expressed protein, serine hydroxymethyltransferase, was confirmed by enzyme linked immunosorbent assay and immunohistochemistry. These results strongly support the idea that the vegetative phase change and the floral transition are regulated independently during developmental process.     

An assessment of morphogenetic fluctuation during reproductive phase change in Arabidopsis

Background and Aims

Reproductive phase change in Arabidopsis thaliana is characterized by two transitions in phytomer identity, the differentiation of the first elongate internode (bolting transition) and of the first flower (floral transition). An evaluation of the dynamics of these transitions was sought by examining the precision of the corresponding phytomer identity changes.

Methods

The length of the first elongate internode and the frequency of chimeric inflorescence structures, e.g. paraclades not subtended by a leaf (no-leaf/paraclades) and flowers subtended by a bract (bract/flowers), were measured in the Wassilewskija (Ws) accession and 47 early flowering mutants under a wide range of photoperiods. The impact of photoperiodic perturbations applied to Ws plants at different times of development was also evaluated.

Key Results

In Ws, both types of characters were remarkably constant across photoperiods in spite of a high degree of interindividual variability. Bract/flowers were not normally produced in Ws, but they were observed in conditions that suggest enhanced light signalling, e.g. in response to continuous light perturbations and in mutants with reduced hypocotyl elongation. In contrast, no-leaf/paraclades were normally present in approx. 20 % of Ws plants, and their frequency was increased in conditions that suggest reduced light signalling, e.g. in mutants with altered specification of long-day responses. The length of the first elongate internode was unrelated to the rate of stem elongation and to the regulation of reproductive phase change.

Conclusions

Bract/flowers and no-leaf/paraclades corresponded to opposite effects on the floral transition that reflected different dynamics of progression to flowering. In contrast, the length of the first elongate internode was only indirectly related to the regulation of reproductive phase change and was mainly dependent on global morphogenetic constraints. This paper proposes that morphogenetic variability could be used to identify critical phases of development and characterize the canalization of developmental patterns.     

Effect of partial defoliation during the vegetative phase on subsequent growth and grain yield of maize

Maize was grown in two densities, 2–47 or 4–94 plants m^-2, and the following treatments imposed: untreated, plants partly defoliated 51 days after sowing, and alternate plants in a row partly defoliated 44 days after sowing. Plants flowered about 82 days after sowing. Leaf area was decreased by 60–64% by defoliation on day 51. Defoliation resulted in decreases in grain yield and grain number of 6–17%, though when alternate plants were defoliated in the higher density there was a substantial decrease in yield and number of grains in defoliated plants, which was largely offset by an increase in adjacent intact plants. When plants were defoliated on day 51 subsequent growth in leaf area was similar to, and that in leaf weight nearly as large as that in untreated plants, while increase in stem weight was substantially less than in untreated plants. By the time of flowering untreated and defoliated plots differed by c. 30% in leaf area. Increments of dry matter after flowering differed by c. 15% between untreated and defoliated plots. The fraction of these increments which entered the grain was c. 90% in both untreated and defoliated plots. When alternate plants in the row were partly defoliated on day 44 their subsequent increase in leaf area was probably 5–16% less than that of the adjacent intact plants. Increments of dry matter after flowering of plots with alternate plants defoliated were 93–95 % of those of untreated plots; leaf efficiency after flowering was slightly greater than in untreated plots. The fraction of the dry matter increment after flowering which entered the grain was c. 88 % in both intact and defoliated plants of the small density, but was 94% in intact plants and 86% in defoliated plants of the large density.