Cassava storage roots are among the most important root crops worldwide and represent one of the most consumed staple foods in Sub-Saharan Africa. The vegetatively propagated tropical shrub can form many starchy tuberous roots from its stem. These storage roots are formed through the activation of secondary root growth processes. However, the underlying genetic regulation of storage root development is largely unknown. Here we report on distinct structural and transcriptional changes occurring during the early phases of storage root development. A pronounced increase in auxin-related transcripts and the transcriptional activation of secondary growth factors, as well as a decrease in gibberellin-related transcripts was observed during the early stages of secondary root growth. This was accompanied by increased cell wall biosynthesis, increased most notably during the initial xylem expansion within the root vasculature. Starch storage metabolism was activated only after the formation of the vascular cambium. The formation of non-lignified xylem parenchyma cells and the activation of starch storage metabolism coincided with increased expression of the KNOX/BEL genes KNAT1, PENNYWISE and POUND-FOOLISH, indicating their importance for proper xylem parenchyma function.

The enzyme phytochelatin synthase (PCS) has long been studied with regard to its role in metal(loid) detoxification in several organisms, i.e., plants, yeasts, and nematodes. It is in fact widely recognized that PCS detoxifies a number of heavy metals by catalyzing the formation of thiol-rich oligomers, namely phytochelatins, from glutathione and related peptides. However, recent investigations have highlighted other possible roles played by the PCS enzyme in the plant cell, e.g., the control of pathogen-triggered callose deposition. In order to examine novel aspects of Arabidopsis thaliana PCS1 (AtPCS1) functions and to elucidate its possible roles in the secondary metabolism, metabolomic data of A. thaliana wild-type and cad1-3 mutant were compared, the latter lacking AtPCS1. HPLC-ESI-MS analysis showed differences in the relative levels of metabolites from the glucosinolate and phenylpropanoid pathways between cad1-3 and wild-type plants. Specifically, in control (Cd-untreated) plants, higher levels of 4-methoxy-indol-3-ylmethylglucosinolate were found in cad1-3 plants vs. wild-type. Moreover, the cad1-3 mutant showed to be impaired in the deposit of callose after Cd exposure, suggesting that AtPCS1 protects the plant against the toxicity of heavy metals not only by synthesizing PCs, but also by contributing to callose deposition. In line with the contribution of callose in counteracting Cd toxicity, we found that another callose-defective mutant, pen2-1, was more sensitive to high concentrations of Cd than wild-type plants. Moreover, cad1-3 plants were more susceptible than wild-type to the hemibiotrophic bacterial pathogen Pseudomonas syringae. The metabolome also revealed differences in the relative levels of hydroxycinnamic acids and flavonols,with consequences on cell wall properties and auxin content, respectively. First, increased lignification in the cad1-3 stems was found, probably aimed at counteracting the entry of Cd into the inner tissues. Second, in cad1-3 shoots, increased relative levels of kaempferol 3,7 dirhamnoside and quercetin hexoside rhamnoside were detected. These flavonols are endogenous inhibitors of auxin transport in planta; auxin levels in both roots and shoots of the cad1-3 mutant were in fact lower than those of the wild-type. Overall, our data highlight novel aspects of AtPCS1 functions in A. thaliana.

The involvement of auxin, abscisic acid (ABA), polyamines (PAs), and proline in adaptation to long-term exposure of woody plants to high levels of heavy metals in soil has received scant attention, even in poplar which is a good candidate for phytoremediation of metal-polluted soils and is regarded as a model for basic research in tree species. Three poplar clones (M1, PE19/66, and B229) were comparatively analyzed in a pot experiment for their responses to 300 mg kg-1 Cu(NO3)2 at morphological, physiological, and biochemical levels. After 4 months, despite the prevalent accumulation of Cu in roots, where the metal reached potentially toxic concentrations, the three clones showed distinct Cu accumulation and translocation capacities, whereas they did not display evident toxicity symptoms or growth inhibition. Several protective mechanisms, namely decreased photosynthetic functionality, enhanced guaiacol peroxidase (GPOD) activity, and accumulation of proline and PAs, were differentially activated in Cu-treated plants in an organ- and clone-specific manner. Overall, a positive relationship between root Cu concentration with GPOD, proline, and PAs was observed. In M1, higher Cu accumulation in roots and leaves compared with other clones was reflected in stimulation of GPOD activity in both organs and in enhanced proline, and PA levels. In PE19/66, these responses were observed only in roots concomitant with high Cu accumulation. Clone B229 accumulated very low amounts of Cu, therefore, these defense responses were attenuated compared with other clones. Enhanced ABA concentrations in response to Cu were observed in PE19/66 and B229; this was likely responsible for stomatal limitation of photosynthesis in PE19/66, whereas in B229 this effect may have been counteracted by increased IAA. Essentially unchanged leaf auxin levels under Cu stress may account for the lack of shoot growth inhibition observed in all three clones; B229 was the only clone that displayed Cu-induced IAA accumulation in roots. Results are discussed in terms of clone-specific adaptive mechanisms to Cu stress in tolerant poplars.

Rhizobacteria with plant growth promoting ability exist in association with plant roots and ameliorate over all plant development and yield. Numerous species of rhizobacteria have been identified with plant growth promoting ability, which can be attributed to multiple microbial characteristics. In the current study rhizobacterial isolates with best plant growth promotion traits were subjected to screening for plant growth promotion under axenic condition. The results of lab assays revealed that out of five rhizobacterial isolates three of bacterial isolate were Gram -ve and two of them were Gram +ve bacterial group. All isolates found positive for the auxin production and ACC-demainase activity. The isolate HS9 showed highest ACC activity (331 ?-ketobutyrate nmol mg-1 biomass hr-1) and auxin production (3.85 without L-TRP). PGPR increase plant growth by reducing the ethylene release and its inhibitory effects, the role of isolates to decrease ethylene effects was affirmed via classical triple response assay on velvet bean. Furthermore, isolate were assessed for resistance test, three efficient strains (G9, HS9 and H38) exhibited antibiotic resistance for streptomycin, kanamycin and rifampicin at100 mg L-1in TSB medium. For the purpose of co-inoculation, all three isolates showed positive relation to grow together. The results concluded that rhizobacteria selected from rain fed areas were found effective to improve plant growth with their multiple growth enhancing traits. Therefore, PGPR with various characteristics could be a better option for inoculation and co-inoculation to improve plant growth in well watered and water stressed environment.

The homeodomain-leucine zipper (HD-Zip) class of transcription factors is unique to plants. HD-Zip proteins bind to DNA exclusively as dimers recognizing dyad symmetric sequences and act as positive or negative regulators of gene expression. On the basis of sequence homology in the HD-Zip DNA-binding domain, HD-Zip proteins have been grouped into four families (HD-Zip I-IV). Each HD-Zip family can be further divided into subfamilies containing paralogous genes that have arisen through genome duplication. Remarkably, all the members of the HD-Zip Ily and -6 clades are regulated by light quality changes that induce in the majority of the angiosperms the shade-avoidance response, a process regulated at multiple levels by auxin. Intriguingly, it has recently emerged that, apart from their function in shade avoidance, the HD-Zip Ily and -6 transcription factors control several auxin-regulated developmental processes, including apical embryo patterning, lateral organ polarity, and gynoecium development, in a white-light environment. This review presents recent advances in our understanding of HD-Zip ll protein function in plant development, with particular emphasis on the impact of loss-of-function HD-Zip II mutations on auxin distribution and response. The review also describes evidence demonstrating that HD-Zip By and -6 genes are directly and positively regulated by HD-Zip III transcription factors, primary determinants of apical shoot development, known to control the expression of several auxin biosynthesis, transport, and response genes. Finally, the interplay between HD-Zip II and III transcription factors in embryo apical patterning and organ polarity is discussed.

The Arabidopsis genome encodes ten Homeodomain-Leucine zipper (HD-Zip) II proteins. ARABIDOPSIS THALIANA HOMEOBOX 2 (ATHB2), HOMEOBOX ARABIDOPSIS THALIANA 1 (HAT1), HAT2, HAT3 and ATHB4 are regulated by changes in the red/far red light ratio that induce shade avoidance in most of the angiosperms. Here, we show that progressive loss of HAT3, ATHB4 and ATHB2 activity causes developmental defects from embryogenesis onwards in white light. Cotyledon development and number are altered in hat3 athb4 embryos, and these defects correlate with changes in auxin distribution and response. athb2 gain-of-function mutation and ATHB2 expression driven by its promoter in hat3 athb4 result in significant attenuation of phenotypes, thus demonstrating that ATHB2 is functionally redundant to HAT3 and ATHB4. In analogy to loss-of-function mutations in HD-Zip III genes, loss of HAT3 and ATHB4 results in organ polarity defects, whereas triple hat3 athb4 athb2 mutants develop one or two radialized cotyledons and lack an active shoot apical meristem (SAM). Consistent with overlapping expression pattern of HD-Zip II and HD-Zip III gene family members, bilateral symmetry and SAM defects are enhanced when hat3 athb4 is combined with mutations in PHABULOSA (PHB), PHAVOLUTA (PHV) or REVOLUTA (REV). Finally, we show that ATHB2 is part of a complex regulatory circuit directly involving both HD-Zip II and HD-Zip III proteins. Taken together, our study provides evidence that a genetic system consisting of HD-Zip II and HDZip III genes cooperates in establishing bilateral symmetry and patterning along the adaxial-abaxial axis in the embryo as well as in controlling SAM activity. © 2013 Published by The Company of Biologists Ltd.

The Arabidopsis genome encodes 10 Homeodomain-Leucine Zipper (HD-Zip) II transcription factors that can be subdivided into 4 clades (alfa-delta). All the gamma (ARABIDOPSIS THALIANA HOMEOBOX 2 [ATHB2], HOMEOBOX ARABIDOPSIS THALIANA 1 [HAT1], HAT2) and delta (HAT3, ATHB4) genes are regulated by light quality changes (Low Red [R]/Far-Red [FR]) that induce the shade avoidance response in most of the angiosperms. HD-Zip II gamma and HD-Zip II delta transcription factors function as positive regulators of shade avoidance, and there is evidence that at least ATHB2 is directly positively regulated by the basic Helix-Loop-Helix (bHLH) proteins PHYTO CHROM E INT ERA CTIN G FACTOR 4 (PIF4) and PIF5. Recent evidence demonstrate that, in addition to their function in shade avoidance, HD-Zip II gamma and HD-Zip II delta proteins play an essential role in plant development from embryogenesis onwards in a white light environment. © 2013 Landes Bioscience.

The radial growth of plant stem is based on the development of cribro-vascular cambium tissues. It affects the transport efficiency of water, mineral nutrients and photoassimilates and, ultimately, also plant height. The rate of cambial cell divisions for the assembly of new xylem and phloem tissue primordia and the rate of differentiation of the primordia into mature tissues determine the amount of biomass produced and, in the case of woody species, the wood quality. These complex physiological processes proceed at a rate which depends on several factors, acting at various levels: growth regulators, resource availability and environmental factors. Several hormonal signals and, more recently, further regulatory molecules, have been shown to be involved in the induction and maintenance of cambium and the formation of secondary vascular tissues. The control of xylem cell patterning is of particular interest, because it determines the diameter of xylem ves- sels, which is central to the efficiency of water and nutrient transport from roots to leaves through the stem and may strongly influence the growth in height of the tree. Increasing scientific evidence have proved the role of other hormones in cambial cell activities and the study of the hormonal signals and their crosstalking in cambial cells may foster our understanding of the dynamics of xylo- genesis and of the mechanism of vessel size control along the stem. In this article, the role of the hormonal signals involved in the control of cambium and xylem develop- ment in trees and their crosstalking are reviewed.

With more than 450 species, Passiflora is the most important genus Of the family Passifloraceae. It comprises; many species grown for their edible fruits, for their high Ornamental value, and further for the therapeutic properties. With their striking exotic flowers, they are of particular interest for the floriculture market. With the aim of evaluating the in vitro propagation of an Italian ornamental hybrid, axillary tendrils of Passiflora "Guglielmo Betto" M. Vecchia (P. incarnata L. x P tucumanensis L) were sterilized and placed in vitro. Direct shoot regeneration was achieved from young tendrils Cultivated oil MS medium containing, either 4.43 mu M 6-benzylaminopurine (BAP) and 11.41 mu M indoleacetic acid (IAA), or 49.20 mu M 6-gamma-gamma-dimethylallylaminopurine (2iP) and 2.68 mu M alpha-naphthalene acetic acid (NAA), respectively. In vitro shoot Multiplication, rooting, and regenerated plant acclimatization protocols were established.

In the present paper we report on the effects of the insertion of the Agrobacterium rhizogenes rolC gene in the tomato (Solanum lycopersicum L., formerly Lycopersicon esculentum Mill.) cultivar Tondino. Several transgenic lines were successfully obtained, between which two clones, rolC1 and rolC3, were chosen for the analysis of morpho-productive traits as well as of the endogenous levels of auxin and abscisic acid. Consistent with the known phenotypic effect of this gene, the transformed tomato plants were significantly shorter than the corresponding controls. On the other hand, even if yield was not affected by the transformation in terms of average number of fruits produced, fruit weight was significantly lower in the transgenics with respect to the controls. Therefore, insertion of the rolC gene does not lead to an improvement in plant productivity. Furthermore, we have observed alterations in the hormonal levels in the shoot apices of the transgenic plants. In fact, quantifications of free and conjugated forms of indole-3-acetic acid (IAA) indicated a significant reduction of IAA levels in the shoot apical region of the transgenic clone rolC3, in comparison with both the control and the clone rolC1. Abscisic acid (ABA) concentration on the other hand was unchanged in the transgenics compared to the controls, but significantly lower in rolC3 with respect to rolC1 plants. The resulting ABA/IAA ratio was higher in both transgenic clones compared to the untransformed plants, indicating that the rolC gene affects the balance between these hormones in transformed tomato plants.

Plant growth regulators are involved in the control of potato (Solanum tuberosum) tuber dormancy. Evidence concerning the role of IAA is controversial; we therefore investigated its role by analyzing two cultivars with varying lengths of dormancy. We examined the time course of free and conjugated IAA in tuber tissue isolates from the final stages of tuber growth to the end of dormancy, the distribution of free IAA in tuber tissues by in situ analysis, and the biosynthesis of the hormone by feeding experiments. The time course of free IAA showed marked differences between the examined cultivars, although the concentration of the auxin generally was the highest at the early stages of tuber dormancy. Immunodetection showed a similar pattern of IAA distribution in both genotypes: in dormant buds from freshly harvested tubers, the free hormone accumulated mostly in apical meristem, leaf and lateral bud primordia, and differentiating vascular tissues underlying the apical meristem, while at the end of the storage period only axillary bud primordia from growing buds displayed appreciable auxin levels. Feeding experiments indicated that changes in IAA biosynthesis rate were a major cause of auxin variation in buds. In both cultivars, dormancy apparently ceased when free IAA fell below a threshold value. Despite this, our data led us to conclude that IAA would not be directly responsible for inhibiting sprouting. Instead, auxin might shorten dormancy, in a cultivar-dependent manner, by enhancing early developmental processes in buds, ultimately leading to dormancy termination.

The paper deals with the effect of sugars on LeEXPA2 expression in tomato (Lycopersicon esculentum) hypocotyl segments, which is a well studied system for analysis of cell elongation. We tested the interactions between sugars and different plant hormones classically known to be involved in plant growth. We found the induction of LeEXPA2 transcript accumulation to be positively affected by the presence of sucrose and other metabolizable sugars. The effect mediated by sorbitol and by a non-metabolizable glucose analogue (3-O-methylglucose) is lower, while the structural analogue of sucrose, turanose, leads to any auxin-induced increase in LeEXPA2 transcript abundance.

Auxin plays a key role in lateral root formation, but the signaling pathway for this process is poorly understood. We show here that NAC1, a new member of the NAC family, is induced by auxin and mediates auxin signaling to promote lateral toot development. NAC1 is a transcription activator consisting of an N-terminal conserved NAG-domain that binds to DNA and a C-terminal activation domain. This faster activates the expression of two downstream auxin-responsive genes, DBP and AIR3. Transgenic plants expressing sense or antisense NAC1: cDNA show an increase or reduction of lateral roots, respectively. finally, TIR1-induced lateral root development is blocked by expression of antisense NAC1 cDNA, and NAC1 overexpression can restore lateral root formation in the auxin-response mutant tir1, indicating that NAC1 acts downstream of TIR1.

The level of auxin - both natural and synthetic -- in the medium has a strong effect on the level of 5-methyl-cytosine in the DNA of carrot cells in culture. This level may vary from approximately 15% to 70% of total cytosine without apparent effects on growth rate and cell morphology. No effect was seen with cytokinin. During somatic embryogenesis, in the absence of hormones, variations were seen in the level of methylation according to a characteristic pattern. If hypomethylation is induced with drugs such as azacytidine, ethionine or ethoxy-carbonyl-pyrimidine, embryogenesis is immediately blocked. A mutant was isolated which is resistant to the action of hypomethylating drugs. It shows variations in the methylation pattern and variations in indole-acetic acid metabolism. In addition its regeneration is often associated with the production of tumors.

We have investigated the relative role of auxin and of Agrobacterium rhizogenes T-DNA in the induction of hairy roots. By infecting carrot discs with suitably constructed bacterial strains containing different T-DNA complements, we have shown that both auxin and the presence of T-DNA in the carrot cells are required for root growth on the discs. Auxin added alone or in combination with cytokinin is not sufficient to induce rooting on uninfected discs. Also cells transformed by T-DNA containing only auxin synthetic genes very rarely differentiate into roots. On the other hand auxin is necessary for hairy root induction since A. rhizogenes devoid of T-DNA-borne auxin genes is not capable of eliciting symptoms in the absence of hormone. Auxin is not required for either T-DNA transfer or T-DNA expression in the transformed host. Cells infected in the absence of auxin, which do not respond by rooting, do contain T-DNA whose expression is shown by the synthesis of hairy root opines; subsequent addition of auxin to these quiescent transformed cells results in root development. A model for hairy root induction where the action of T-DNA is envisaged as conferring auxin responsiveness to the transformed cells is discussed. © 1987 Springer-Verlag.