A lysine-rich arabinogalactan protein in Arabidopsis is essential for plant growth and development, including cell division and expansion

Arabinogalactan proteins (AGPs), a family of hydroxyproline-rich glycoproteins, occur throughout the plant kingdom. The lysine-rich classical AGP subfamily in Arabidopsis consists of three members, AtAGP17, 18 and 19. In this study, AtAGP19 was examined in terms of its gene expression pattern and function. AtAGP19 mRNA was abundant in stems, with moderate levels in flowers and roots and low levels in leaves. AtAGP19 promoter-controlled GUS activity was high in the vasculature of leaves, roots, stems and flowers, as well as styles and siliques. A null T-DNA knockout mutant of AtAGP19 was obtained and compared to wild-type (WT) plants. The atagp19 mutant had: (i) smaller, rounder and flatter rosette leaves, (ii) lighter-green leaves containing less chlorophyll, (iii) delayed growth, (iv) shorter hypocotyls and inflorescence stems, and (v) fewer siliques and less seed production. Several abnormalities in cell size, number, shape and packing were also observed in the mutant. Complementation of this pleiotropic mutant with the WT AtAGP19 gene restored the WT phenotypes and confirmed that AtAGP19 functions in various aspects of plant growth and development, including cell division and expansion, leaf development and reproduction.     

Characterization and genetic analysis of a lettuce (Lactuca sativa L.) mutant,weary, that exhibits reduced gravitropic response in hypocotyls and inflorescence stems

A lettuce (Lactuca sativa L.) mutant that exhibits a procumbent growth habit was identified and characterized. In two wild type (WT) genetic backgrounds, segregation patterns revealed that the mutant phenotype was controlled by a recessive allele at a single locus, which was designated weary. Hypocotyls and inflorescence stems of plants homozygous for the weary allele exhibited reduced gravitropic responses compared with WT plants, but roots exhibited normal gravitropism. Microscopic analysis revealed differences in the radial distribution of amyloplasts in hypocotyl and inflorescence stem cells of weary and WT plants. Amyloplasts occurred in a single layer of endodermal cells in WT hypocotyls and inflorescence stems. By contrast, amyloplasts were observed in several layers of cortical cells in weary hypocotyls, and weary inflorescence stem cells lacked amyloplasts entirely. These results are consistent with the proposed role of sedimenting amyloplasts in shoot gravitropism of higher plants. The phenotype associated with the weary mutant is similar to that described for the Arabidopsis mutant sgr1/scr, which is defective in radial patterning and gravitropism.     

Cloning of an Arabidopsis patatin-like gene, STURDY, by activation T-DNA tagging

Activation T-DNA tagging can generate dominant gain-of-function mutants by overexpression of a particular endogenous gene. We identified an activation-tagged mutant, sturdy, exhibiting a stiff inflorescence stem, thicker leaves, shorter siliques, larger seeds, round-shaped flowers, and delayed growth. It is most important that unlike its wild-type counterpart, this mutant is less prone to lodging. Cloning of STURDY revealed that in sturdy, there is an open reading frame containing a single intron encoding a patatin-like homolog. The T-DNA is inserted into the 3' region of the second exon. The mutant phenotype was shown to be the result of overexpression of STURDY by mRNA analysis and transgenic studies. Preliminary histological studies have revealed an increase in cell number in the inflorescence stem of mutant plants; however, additional studies are needed to better understand the overexpression phenotype.     

Stamenless, a tomato mutant with homeotic conversions in petals and stamens

A tomato (Lycopersicon esculentum Mill.) monogenic semidominant mutation, stamenless (sl), which results in homeotic conversions in two adjacent floral whorls, was studied. When grown at standard temperature, flowers of sl/sl plants showed sepaloid petals in the second whorl and strong transformation of stamens to carpels in whorl three. These transformed carpels were fused with each other and with the genuine carpels in the fourth whorl to form a unique gynoecium. The mutation is semidominant since heterozygous plants showed a phenotype intermediate between that of the wild type (WT) and that of homozygous mutant plants, with nearly WT petals but with feminized stamens bearing naked ovules on the base of their adaxial face. The initiation and position of organ primordia in sl/sl flowers were not altered when compared with WT primordia although development of organ primordia in the second and third whorls deviated from WT at an early stage as observed by scanning electron microscopy. The mutant phenotype is temperature sensitive and when sl/sl plants were cultured at low temperature, the morphology of some flowers resembled that of the WT. This reversion of the mutant phenotype is also induced by treatment of young sl/sl plants with gibberellic acid, providing evidence that gibberellin synthesis or sensitivity could mediate the effect of low temperature on the mutant phenotype. Southern blot analyses using a Deficiens-homologous gene from Solanum tuberosum as a probe showed a restriction-fragment-length polymorphism (RFLP) linked to the sl mutation. This result indicates that the mutation affects a Deficiens-like gene that controls the identity of petals and stamens. Received: 10 December 1998 / Accepted: 29 March 1999     

Deficiency in fatty acid synthase leads to premature cell death and dramatic alterations in plant morphology

An Arabidopsis mosaic death1 (mod1) mutant, which has premature cell death in multiple organs, was isolated. mod1 plants display multiple morphological phenotypes, including chlorotic and curly leaves, distorted siliques, premature senescence of primary inflorescences, reduced fertility, and semidwarfism. The phenotype of the mod1 mutant results from a single nuclear recessive mutation, and the MOD1 gene was isolated by using a map-based cloning approach. The MOD1 gene encodes an enoyl-acyl carrier protein (ACP) reductase, which is a subunit of the fatty acid synthase complex that catalyzes de novo synthesis of fatty acids. An amino acid substitution in the enoyl-ACP reductase of the mod1 mutant causes a marked decrease in its enzymatic activity, impairing fatty acid biosynthesis and decreasing the amount of total lipids in mod1 plants. These results demonstrate that a deficiency in fatty acid biosynthesis has pleiotropic effects on plant growth and development and causes premature cell death.     

Pollen tube and root-hair tip growth is disrupted in a mutant of Arabidopsis thaliana.

The expansion of both root hairs and pollen tubes occurs by a process known as tip growth. In this report, an Arabidopsis thaliana mutant (tip1) is described that displays defects in both root-hair and pollen-tube growth. The root hairs of the tip1 mutant plants are shorter than those of the wild-type plants and branched at their base. The tip1 pollen-tube growth defect was identified by the aberrant segregation ratio of phenotypically normal to mutant seeds in siliques from self-pollinated, heterozygous plants. Homozygous mutant seeds are not randomly distributed in the siliques, comprising only 14.4% of the total seeds, 5.3% of the seeds from the bottom half, and 2.2% of the seeds from the bottom quarter of the heterozygous siliques. Studies of pollen-tube growth in vivo showed that mutant pollen tubes grow more slowly than wild-type pollen through the transmitting tissue of wild-type flowers. Cosegregation studies indicate that the root-hair and pollen-tube defects are caused by the same genetic lesion. Based on these findings, the TIP1 gene is likely to encode a product involved in a fundamental aspect of tip growth in plant cells.     

A Mutation in the Arabidopsis TFL1 Gene Affects Inflorescence Meristem Development

We present the initial phenotypic characterization of an Arabidopsis mutation, terminal flower 1-1 (tfl1-1), that identifies a new genetic locus, TFL1. The tfl1-1 mutation causes early flowering and limits the development of the normally indeterminate inflorescence by promoting the formation of a terminal floral meristem. Inflorescence development in mutant plants often terminates with a compound floral structure consisting of the terminal flower and one or two subtending lateral flowers. The distal-most flowers frequently contain chimeric floral organs. Light microscopic examination shows no structural aberrations in the vegetative meristem or in the inflorescence meristem before the formation of floral buttresses. The wild-type appearance of lateral flowers and observations of double mutant combinations of tfl1-1 with the floral morphogenesis mutations apetala 1-1 (ap1-1), ap2-1, and agamous (ag) suggest that the tfl1-1 mutation does not affect normal floral meristems. Secondary flower formation usually associated with the ap1-1 mutation is suppressed in the terminal flower, but not in the lateral flowers, of tfl1-1 ap1-1 double mutants. Our results suggest that tfl1-1 perturbs the establishment and maintenance of the inflorescence meristem. The mutation lies on the top arm of chromosome 5 approximately 2.8 centimorgans from the restriction fragment length polymorphism marker 217.     

Dominant-negative receptor uncovers redundancy in the Arabidopsis ERECTA Leucine-rich repeat receptor-like kinase signaling pathway that regulates organ shape

Arabidopsis ERECTA, a Leu-rich repeat receptor-like Ser/Thr kinase (LRR-RLK), regulates organ shape and inflorescence architecture. Here, we show that a truncated ERECTA protein that lacks the cytoplasmic kinase domain (DeltaKinase) confers dominant-negative effects when expressed under the control of the native ERECTA promoter and terminator. Transgenic plants expressing DeltaKinase displayed phenotypes, including compact inflorescence and short, blunt siliques, that are characteristic of loss-of-function erecta mutant plants. The DeltaKinase fragment migrated as a stable approximately 400-kD protein complex in the complete absence of the endogenous ERECTA protein and significantly exaggerated the growth defects of the null erecta plants. A functional LRR domain of DeltaKinase was required for dominant-negative effects. Accumulation of DeltaKinase did not interfere with another LRR-RLK signaling pathway (CLAVATA1), which operates in the same cells as ERECTA but has a distinct biological function. Both the erecta mutation and DeltaKinase expression conferred a lesser number of large, disorganized, and expanded cortex cells, which are associated with an increased level of somatic endoploidy. These findings suggest that functionally redundant RLK signaling pathways, including ERECTA, are required to fine-tune the proliferation and growth of cells in the same tissue type during Arabidopsis organogenesis.     

The Arabidopsis compact inflorescence genes: phase-specific growth regulation and the determination of inflorescence architecture

During their life cycle, higher plants pass through a series of growth phases that are characterized by the production of morphologically distinct vegetative and reproductive organs and by different growth patterns. Three major phases have been described in Arabidopsis: juvenile vegetative, adult vegetative, and reproductive. In this report we describe a novel, phase-specific mutant in Arabidopsis, compact inflorescence (cif). The most apparent aspect of the cif phenotype is a strong reduction in the elongation of internodes in the inflorescence, resulting in the formation of a floral cluster at the apical end of all reproductive shoots. Elongation and expansion of adult vegetative rosette leaves are also compromised in mutant plants. The onset of the cif trait correlates closely with morphological changes marking the phase transition from juvenile to adult, and mutant plants produce normal flowers and are fully fertile. Hence the cif phenotype appears to be adult vegetative phase-specific. Histological sections of mutant inflorescence internodes indicate normal tissue specification, but reduced cell elongation compared to wild-type. compact inflorescence is inherited as a two-gene trait involving the action of a recessive and a dominant locus. These two cif genes appear to be key components of a growth regulatory pathway that is closely linked to phase change, and specifies critical aspects of plant growth and architecture including inflorescence internode length.     

The Evergreen gene is essential for flower initiation in carnation

One of the leading cut-flower crops in the world, the greenhouse carnation (Dianthus caryophyllus), has been subjected to intense breeding efforts for the past few hundred years. As an ornamental crop, flowering and flower architecture are major breeding targets that are constantly in demand. In an ongoing breeding program aimed at improving these characteristics, two mutants heterozygous for a mutation in a gene termed evergreen (e) were identified. In these mutants, spike-like clusters of bracteoles subtend each flower. Genetic analysis of the mutants confirmed the semidominant nature of this nuclear mutation and that the two original mutants were allelic at the evergreen locus. In homozygous mutant plants, a more severe phenotype was observed. Flower formation was completely blocked and spikelike clusters of bracteoles did not subtend any flowers. Morphological characterization of mutant plants revealed that vegetative growth and inflorescence structure are not affected by the mutant allele. In plants heterozygous for the evergreen mutation, fertility, petal and pistil length, calyx diameter, and stamen number were not affected. However, flowers from these heterozygous plants had a reduced number of petals, suggesting an intriguing link between evergreen and the double flower (d) gene that determines petal number in carnation. The control by evergreen of bracteole formation, floral meristem initiation, and petal number in carnation is discussed in comparison to the recessive leafy (lfy) and floricaula (flo) mutants of Arabidopsis and Antirrhinum, respectively.     

Formation of corymb-like inflorescences due to delay in bolting and flower development in the corymbosa2 mutant of Arabidopsis

Among the wild-type ecotypes of Arabidopsis thaliana whose shape of inflorescence is categorized as raceme, the ecotype Landsberg harboring the erecta (er) mutation shows a corymb-like inflorescence, namely, a compact inflorescence with a flattened arrangement of flower buds at the tip. The fact that the ER gene encodes a receptor-like protein kinase implies the presence of a signaling cascade responsible for the inflorescence morphology of flowering plants. We report here the characterization of another mutant with a corymb-like inflorescence, named corymbosa2 (crm2), and the isolation of the CRM2 gene. While the er mutation causes a severe reduction in the length of pedicels, the crm2 mutation results in a significant delay in the initiation of internode elongation and in the development of flowers, despite having little effect on the timing of floral induction. Consequently, the number of flower buds is apparently increased at the tip of crm2 inflorescence. The crm2 er double mutant shows an additive phenotype. These results suggest that CRM2 and ER may act in different ways to generate wild-type inflorescence. The CRM2 gene was isolated by positional cloning and appears to encode a polypeptide with no significant homology to known sequences.     

Stems of the <Emphasis Type="Italic">Arabidopsis pin1-1</Emphasis> mutant are not deficient in free indole-3-acetic acid

The pin1-1 mutant of Arabidopsis thaliana has been pivotal for studies on auxin transport and on the role of auxin in plant development. It was reported previously that when whole shoots were analysed, levels of the major auxin, indole-3-acetic acid (IAA) were dramatically reduced in the mutant, compared with the WT (Okada et al. 1991). The cloning of PIN1, however, provided evidence that this gene encodes a facilitator of auxin efflux, raising the question of how the pin1-1 mutation might reduce overall IAA levels as well as IAA transport. We therefore re-examined IAA levels in individual parts of pin1-1 and WT plants, focusing on inflorescence stems. Our data show that there is in fact no systemic IAA deficiency in the mutant. The previously reported difference between mutant and WT may have been due to the inclusion of reproductive structures in the WT harvest: we show here that the inflorescence itself contains high levels of IAA. We reconcile the normal IAA levels of pin1-1 inflorescence stems with their (previously-reported) reduced ability to transport IAA by presenting evidence that the auxin in mutant stems is not imported from their apical portion. Our data also indicate that levels of another auxin, indole-3-butyric acid (IBA), are very low in stems of the genotypes used in this study.     

Arabidopsis thickvein mutation affects vein thickness and organ vascularization, and resides in a provascular cell-specific spermine synthase involved in vein definition and in polar auxin transport

Polar auxin transport has been implicated in the induction of vascular tissue and in the definition of vein positions. Leaves treated with chemical inhibitors of polar auxin transport exhibited vascular phenotypes that include increased vein thickness and vascularization. We describe a recessive mutant, thickvein (tkv), which develops thicker veins in leaves and in inflorescence stems. The increased vein thickness is attributable to an increased number of vascular cells. Mutant plants have smaller leaves and shorter inflorescence stems, and this reduction in organ size and height is accompanied by an increase in organ vascularization, which appears to be attributable to an increase in the recruitment of cells into veins. Furthermore, although floral development is normal, auxin transport in the inflorescence stem is significantly reduced in the mutant, suggesting that the defect in auxin transport is responsible for the vascular phenotypes. In the primary root, the veins appear morphologically normal, but root growth in the tkv mutant is hypersensitive to exogenous cytokinin. The tkv mutation was found to reside in the ACL5 gene, which encodes a spermine synthase and whose expression is specific to provascular cells. We propose that ACL5/TKV is involved in vein definition (defining the boundaries between veins and nonvein regions) and in polar auxin transport, and that polyamines are involved in this process.     

Changes in growth and photosynthetic characteristics of Ocimum sanctum under growth regulator treatments

A field experiment was conducted to investigate the effect of growth regulators on growth characteristics such as root length, shoot length, total leaf area, number of inflorescence per plant, number of flower per inflorescence, whole plant fresh weight and whole plant dry weight. Photosynthetic characteristics were also analyzed based on the same experiment. For this, various photosynthetic pigment contents such as chlorophyll, carotenoid, anthocyanin and xanthophyll content were calculated. The conventional growth regulator abscisic acid (ABA) and non-conventional growth regulator triazole compound paclobutrazol (PBZ) were used. Root length increased due to growth regulator treatment, but shoot length decreased. Leaf area was decreased due to growth regulator treatment. The number of inflorescence increased in ABA treated plants, but it was decreased in PBZ treated plants. In ABA treated plants, the number of flowers per inflorescence was increased. In PBZ treated plants the number of inflorescence was reduced. The whole plant fresh weight (FW) and dry weight (DW) were increased in ABA and PBZ treated plants. There was an increase in chlorophyll content in growth regulator treated plants compared to control, and it was more in PBZ treated plants. The carotenoid content was also increased in ABA and PBZ treated plants.     

Flavonoids act as negative regulators of auxin transport in vivo in arabidopsis

Polar transport of the plant hormone auxin controls many aspects of plant growth and development. A number of synthetic compounds have been shown to block the process of auxin transport by inhibition of the auxin efflux carrier complex. These synthetic auxin transport inhibitors may act by mimicking endogenous molecules. Flavonoids, a class of secondary plant metabolic compounds, have been suggested to be auxin transport inhibitors based on their in vitro activity. The hypothesis that flavonoids regulate auxin transport in vivo was tested in Arabidopsis by comparing wild-type (WT) and transparent testa (tt4) plants with a mutation in the gene encoding the first enzyme in flavonoid biosynthesis, chalcone synthase. In a comparison between tt4 and WT plants, phenotypic differences were observed, including three times as many secondary inflorescence stems, reduced plant height, decreased stem diameter, and increased secondary root development. Growth of WT Arabidopsis plants on naringenin, a biosynthetic precursor to those flavonoids with auxin transport inhibitor activity in vitro, leads to a reduction in root growth and gravitropism, similar to the effects of synthetic auxin transport inhibitors. Analyses of auxin transport in the inflorescence and hypocotyl of independent tt4 alleles indicate that auxin transport is elevated in plants with a tt4 mutation. In hypocotyls of tt4, this elevated transport is reversed when flavonoids are synthesized by growth of plants on the flavonoid precursor, naringenin. These results are consistent with a role for flavonoids as endogenous regulators of auxin transport.     

A DEAD box RNA helicase is essential for mRNA export and important for development and stress responses in Arabidopsis

An Arabidopsis thaliana mutant, cryophyte, was isolated and found to have an enhanced cold stress-induction of the master regulator of cold tolerance, C-repeat binding factor 2 (CBF2), and its downstream target genes. The mutant is more tolerant to chilling and freezing stresses but is more sensitive to heat stress. Under warm but not cold growth temperatures, the mutant has a reduced stature and flowers earlier. Under long day conditions, flowering of the mutant is insensitive to vernalization. The mutant is also hypersensitive to the phytohormone abscisic acid. The mutation was found in a DEAD box RNA helicase gene that is identical to the previously identified low expression of osmotically responsive genes 4 (LOS4) locus, which was defined by the los4-1 mutation that reduces cold regulation of CBFs and their target genes and renders Arabidopsis plants chilling sensitive. We show evidence suggesting that the CRYOPHYTE/LOS4 protein may be enriched in the nuclear rim. In situ poly(A) hybridization indicates that the export of poly(A)+ RNAs is blocked in the cryophyte/los4-2 mutant at warm or high temperatures but not at low temperatures, whereas the los4-1 mutation weakens mRNA export at both low and warm temperatures. These results demonstrate an important role of the CRYOPHYTE/LOS4 RNA helicase in mRNA export, plant development, and stress responses.     

DETERMINATE (det) MUTANT OF PISUM SATIVUM (LEGUMINOSAE: PAPILIONOIDEAE) EXHIBITS AN INDETERMINATE GROWTH PATTERN

Inflorescence ontogeny and morphology of the det mutant of Pisum sativum L. were investigated using scanning electron microscopy. This mutation causes the production of a limited number of axillary flowers followed by the formation of an apparent terminal flower slightly offset from the vertical. Our study indicates that the apparent terminal flower arises from an axillary meristem. The terminal meristem senesces and differentiates hairs, forming a rudimentary stub in the same manner as axillary meristems of conventional (Det) and det plants. Thus the dramatic effect of the det gene on inflorescence architecture results from early apical arrest rather than conversion of the terminal meristem to a flower as implied by the symbol det. This mutant will be valuable in elucidating regulation of apical arrest.     

Changes in growth and photosynthetic characteristics of Ocimum sanctum under growth regulator treatments

A field experiment was conducted to investigate the effect of growth regulators on growth characteristics such as root length,shoot length,total leaf area,number of inflorescence per plant,number of flower per inflorescence,whole plant fresh weight and whole plant dry weight.Photosynthetic characteristics were also analyzed based on the same experiment.For this,various photosynthetic pigment contents such as chlorophyll,carotenoid,anthocyanin and xanthophyll content were calculated.The conventional growth regulator abscisic acid (ABA) and non-conventional growth regulator triazole compound paclobutrazol (PBZ) were used.Root length increased due to growth regulator treatment,but shoot length decreased.Leaf area was decreased due to growth regulator treatment.The number of inflorescence increased in ABA treated plants,but it was decreased in PBZ treated plants.In ABA treated plants,the number of flowers per inflorescence was increased.In PBZ treated plants the number of inflorescence was reduced.The whole plant fresh weight (FW) and dry weight (DW) were increased in ABA and PBZ treated plants.There was an increase in chlorophyll content in growth regulator treated plants compared to control,and it was more in PBZ treated plants.The carotenoid content was also increased in ABA and PBZ treated plants.