Modified hormonal balance in rooting-recalcitrant rac mutant tobacco shoots

ABSTRACT

The rooting-recalcitrant rac tobacco mutant has been multiplied in vitro via outgrowth of axillary buds in parallel to the D8 wild-type. The mutant shoots grew at a lower rate and did not root whatever the treatments, whereas the wild-type shoots rooted spontaneously during the culture cycle without auxin treatment. The mutant and wild-type shoots showed similar peroxidase variations along the culture cycle (21 days) but with higher levels of activity for the rac mutant: minimum peroxidase activity occurred at day 14 in whole shoots of both tobacco genotypes, but already at day 7 in the basal parts of the stems (where roots appear) of the wild-type tobacco, while it was delayed in the mutant. Free and conjugated auxin and polyamine levels were also determined in whole shoots and basal parts of the stems. The rac mutant was characterised by higher auxin and polyamine contents. A peak of auxins and polyamines appeared at day 14 in the whole shoots whatever the tobacco genotype. This peak was delayed in the basal parts of the rac stems compared to the wild-type ones. The mutant shoots contained higher levels of benzyladenine and isopentenyladenosine at the end of the culture cycle, whereas zeatin riboside was more abundant in wild-type shoots. In response to increasing concentrations of indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA), only the wild-type shoots responded by an increase in growth rate followed by inhibition at high concentrations. The rac shoot responses were very low or nonexistent. Peroxidase activity was also measured in the basal parts of tobacco stems grown in the presence of IBA. Results suggest growth inhibition related to auxin accumulation, possibly combined with elevated putrescine content. Second, rooting induction seems to take place in both tobacco genotypes; however, the process of root formation is blocked in the mutant. The lack of initiation and expression phases of rooting in relation to auxin content in the mutant is discussed.     

Distribution and survival of overwintering adults of the Dutch elm disease vector, Hylurgopinus rufipes (Coleoptera: Scolytidae), in American elm trees in Manitoba

Abstract 1 The native elm bark beetle, Hylurgopinus rufipes, is the principal vector of Dutch elm disease in Manitoba, Saskatchewan and North Dakota, and disease management measures include applying residual insecticides to the lower part of elm tree trunks where the adult beetles overwinter. 2 In American elm trees in southern Manitoba, we counted entrance holes produced by beetles then felled and dissected trees to determine numbers of tunnels and numbers and survival of overwintering beetles. 3 Densities of entrance holes, tunnels and beetles followed a logistic relationship with tree trunk diameter; densities were near zero at diameters < 10 cm and reached a site‐specific asymptote at diameters > 20 cm. 4 Asymptotic densities of holes, tunnels and beetles in samples from 55 to 190 cm above the ground were, respectively, 22%, 22% and 0.7% of those within 25 cm of the ground. 5 Within the height range 0–190 cm, the proportion of living beetles declined steeply with increasing height. 6 Average density of holes at height 0–25 cm estimated from a sample of several trees of diameter ≥15 cm could be used to predict the asymptotic maximum density of overwintering beetles in the site; predictions of beetle densities for individual trees were not reliable.     

Cytokinins in shoots of the chestnut tree

Five cytokinins, trans-zeatin, 9-β-d-ribosyl-trans-zeatin, O-β-d-glucosyl-trans-zeatin, N⁶-(3-methyl-but-2-enyl)adenine and N⁶-(3-methyl-but-2-enyl)adenosine were identified in shoots of the chestnut tree.     

激素对香椿腋芽增殖生长的效应

在MS基本培养其中附加不同种类的外源激素,对其在香椿芽增殖培养中的效果进行了对比试验,结果表明：单一激素对芽增殖不利,细胞分裂素是导致植株玻璃化的主要因素,只有6-BA和GA3配合使用芽增殖效果最好;GA3对6-BA有明显增效作用,既能促进芽增殖生长,又能防止、减轻玻璃化苗产生。     

Radiocarbon wiggle-dating of elm declines in northwest Denmark and their significance

Four elm declines were found in a pollen diagram from a small lake in northwest Denmark. Matching consecutive radiocarbon dates with the dendro-chronological calibration curve indicated a reservoir effect of 120 years, and dates for the four elm declines were obtained (4530, 4130, 3870, 3410 cal B.C.). The occurrence of apophytes (native plants encouraged by human activities) and increased vegetation diversity during the four elm declines indicates human disturbance. The first and second elm declines coincide with traces of early agriculture in northern Germany. The third and fourth elm declines are contemporary with the transitions to the Early Neolithic and the Middle Neolithic Funnel Beaker Culture in Denmark. The possible influence of outbreaks of elm disease is discussed.     

The role of phytohormones ethylene and auxin in plant-nematode interactions

The phytohormones ethylene and auxin regulate many important processes in plants, including cell differentiation, cell expansion, and responses to abiotic stresses. These hormones also play important roles in many plant-pathogen interactions, including regulation of plant defense responses and symptom development. Sedentary plant-parasitic nematodes, which require the formation of a complex feeding site within the host root, are among the world’s most destructive plant pathogens. Nematode-induced feeding sites show dramatic changes in host cell morphology and gene expression. These changes are likely mediated, at least in part, by phytohormones. In the present review, current knowledge of the roles of ethylene and auxin will be explored in two main areas: the specific role of phytohormones in mediating feeding site development by plant-parasitic nematodes and the general role of phytohormones in affecting the ability of parasitic nematodes to cause disease. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 1, pp. 3–7. This article was presented in original.     

松嫩草原榆树疏林对不同干扰的响应

榆树疏林是中国东北松嫩草原沙地植被演替的顶极群落,其对维持物种多样性,保持水土,涵养水源等都发挥着重要的功能。过去20a间,榆树疏林在人为干扰(农耕和放牧)下,其群落结构和物种组成发生了巨大变化。在1983年和2004年,对松嫩草原不同人为干扰下榆树疏林群落结构和物种组成等进行调查,结果显示:农耕和放牧一方面使松嫩草原榆树疏林群落多物种消失、生物量和多样性显著下降、植被结构简单化;另一方面,也是导致建群种——家榆低矮化,灌木化的主要因素。     

松嫩平原杂类草草甸和榆树疏林草原大油芒种群的年龄结构

对松嫩平原杂类草草甸和榆树疏林草原上大油芒种群的年龄结构及各龄级构件的物质生产力和营养繁殖力进行了研究.结果表明:在2003和2006年中,2个生境的大油芒种群分蘖株均由2～3个龄级组成,根茎由4个龄级组成,分蘖株和根茎均以1a和2a所占比例最大,年龄结构呈增长型或稳定型;分蘖节芽以1a或2a者占优势,根茎顶端芽在芽库中所占比例为29.4%～45.0%,对翌年种群的更新具有重要作用;分蘖株、根茎的物质生产力和营养繁殖力均以1a或2a构件最大,根茎比分蘖节具有更旺盛的营养繁殖力.     

Host responses and metabolic profiles of wood components in Dutch elm hybrids with a contrasting tolerance to Dutch elm disease

Background and Aims

Changes occurring in the macromolecular traits of cell wall components in elm wood following attack by Ophiostoma novo-ulmi, the causative agent of Dutch elm disease (DED), are poorly understood. The purpose of this study was to compare host responses and the metabolic profiles of wood components for two Dutch elm (Ulmus) hybrids, ‘Groeneveld’ (a susceptible clone) and ‘Dodoens’ (a tolerant clone), that have contrasting survival strategies upon infection with the current prevalent strain of DED.

Methods

Ten-year-old plants of the hybrid elms were inoculated with O. novo-ulmi ssp. americana × novo-ulmi. Measurements were made of the content of main cell wall components and extractives, lignin monomer composition, macromolecular traits of cellulose and neutral saccharide composition.

Key Results

Upon infection, medium molecular weight macromolecules of cellulose were degraded in both the susceptible and tolerant elm hybrids, resulting in the occurrence of secondary cell wall ruptures and cracks in the vessels, but rarely in the fibres. The ¹³C nuclear magnetic resonance spectra revealed that loss of crystalline and non-crystalline cellulose regions occurred in parallel. The rate of cellulose degradation was influenced by the syringyl:guaiacyl ratio in lignin. Both hybrids commonly responded to the medium molecular weight cellulose degradation with the biosynthesis of high molecular weight macromolecules of cellulose, resulting in a significant increase in values for the degree of polymerization and polydispersity. Other responses of the hybrids included an increase in lignin content, a decrease in relative proportions of d-glucose, and an increase in proportions of d-xylose. Differential responses between the hybrids were found in the syringyl:guaiacyl ratio in lignin.

Conclusions

In susceptible ‘Groeneveld’ plants, syringyl-rich lignin provided a far greater degree of protection from cellulose degradation than in ‘Dodoens’, but only guaiacyl-rich lignin in ‘Dodoens’ plants was involved in successful defence against the fungus. This finding was confirmed by the associations of vanillin and vanillic acid with the DED-tolerant ‘Dodoens’ plants in a multivariate analysis of wood traits.     

Interaction between day length and phytohormones in the control of potato tuberization in the in vitro culture

We studied the interaction of the day length, cytokinins, and gibberellins in the control of tuberization in potato (Solanum tuberosum L, cv. Desire) plants and derived transgenic plants with the inserted PHYB gene from Arabidopsis encoding the synthesis of phytochrome B apoprotein and put under the control of the 35S CaMV promoter. Plantlets were cultured in vitro on hormone-free MS medium containing 5% sucrose and kinetin (1 mg/l) or/and GA (0.5 and 1.0 mg/l), at long day (LD, a 16-h photoperiod), short day (SD, a 10-h photoperiod), or continuous darkness conditions. The content of cytokinins (Ck, zeatin, and zeatin riboside) in various plant organs was determined by the immunoenzyme method, and GA activity was measured in bioassay with dwarf pea. Potato plant transformation with the PHYB gene enhanced substantially tuber initiation inhibition by LD. Kinetin addition to culture medium enhanced tuberization and reduced Ck content in aboveground shoots and Ck redistribution in the favor of underground organs. GA addition to the culture medium suppressed tuberization and induced Ck accumulation in aboveground organs. We concluded that Ck role in tuberization depends on their predominant localization in above- or underground potato organs. The involvement of Ck and GA in the competitive relations between growing tubers and shoots is considered.     

Involvement of putrescine in the inductive rooting phase of poplar shoots raised in vitro

Micropropagated poplar shoots rooted 100% on a rooting medium (A) containing NAA, but they did not root in the absence of auxin (NA). Putrescine, but not spermidine and spermine, promoted rooting up to 42% when added to the NA medium. Cyclohexylamine (CHA), an inhibitor of spermine synthase, also promoted (up to 36%) rooting in the absence of auxin. The inhibitors of polyamine biosynthesis DFMA (α-difluoromethylarginine) and DFMO (α-difluoromethylomithine), aminoguanidine (AG) and methylglyoxal-bis-guanylhydrazone (MGBG), inhibited rooting when applied in the presence of auxin and had no effect in its absence.
The rooting inductive phase (in the presence of auxin) was determined by periodical transfer of shoots from A to NA medium, and by changes in peroxidase activity, to be 7 h. Putrescine (not spermidine and spermine) accumulated to a maximum during the inductive phase. Both putrescine and CHA promoted rooting on NA medium when applied during the first 7 h. In contrast DFMA and AG inhibited rooting during this period. The results point to the involvement of putrescine and its Δ¹-pyrroline pathway, in the inductive phase of rooting in poplar shoots.     

In vitro root induction in axillary microshoots of Quercus robur L.

Whilst considerable efforts have been made to optimise shoot multiplication and rooting in oak, little attention has been paid to the impact of conditions used for multiplication on subsequent root formation. An optimised technique for rooting of oak microshoots has been developed to assess the effect of cytokinin treatments applied to shoot multiplication cultures on the subsequent rooting of microshoots. We found IBA to be more effective at inducing root formation in microshoots than NAA. Efficient rooting of oak microshoots (80%) was achieved after 35 days on medium supplemented with 1.0 mg litre^-1 IBA. Lower concentrations of IBA reduced the frequency of root formation and significantly increased the time taken for microshoots to form roots. High concentrations of IBA (3.0 mg litre^-1) produced similar rooting frequencies but with significantly increased numbers of roots formed by each microshoot. However, high concentrations of IBA stimulated the production of basal callus. Rooting of microshoots was unaffected by the concentration of BA used during shoot multiplication, although basal callusing was greater in microshoots taken from multiplication medium supplemented with the highest concentration of BA (1.0 mg litre^-1) and rooted on medium supplemented with 3.0 mg litre IBA. Reducing the period of exposure to auxin to 7 days by transferring microshoots to auxin-free medium increased the frequency of root formation (84%), led to more rapid root formation and a reduction in basal callus formation.     

Seasonal changes in wood formation of Ulmus pumila and U. minor and its relation with Dutch elm disease

Elms containing narrow and scattered vessels have been reported to be more resistant to Ophiostoma novo-ulmi (Dutch elm disease pathogen) than elms with large and contiguous vessels. However, recent measurements in Ulmus pumila and U. minor showed a contrary trend. The pin method was applied to 4-yr-old branches of eight clones planted in Madrid. During 2002, radial growth increments and vessel diameters were measured monthly, and beetle trapping was undertaken weekly. U. minor formed larger vessels at the beginning of the season, coinciding with a peak of captured beetles, but, up to June 15, vessels were larger for U. pumila. The number of vessels per group, the transversal area per vessel group, and the mean theoretical hydraulic conductances were significantly higher for U. minor on most dates. Researchers should take into consideration the seasonal changes in vessel size. The results highlight that seasonal variation of vessel diameters and hydraulic parameters, in combination with beetle abundance, are the main factors that could explain the different susceptibility of both elm species to O. novo-ulmi.     

Comparison of effects of bacterial strains differing in their ability to synthesize cytokinins on growth and cytokinin content in wheat plants

The content of cytokinins and pigments together with the morphological parameters and fresh weight were estimated in durum wheat (Triticum durum Desf.) plants 2–4 days after introduction into their rhizosphere of an aliquot of Bacillus suspension using the strains that differed in their ability of producing cytokinins. The experiments were performed under laboratory conditions at the optimum light intensity and mineral nutrition. Inoculation with microorganisms incapable to synthesize cytokinins did not affect the total cytokinin content in the wheat plants, whereas the presence of cytokinin-producing microorganisms in the rhizosphere was accompanied by a considerable increase in the total cytokinin content and the accumulation of individual hormones. On the second day after inoculation, a dramatic increase in zeatin riboside and zeatin O-glucoside contents was observed in the roots, and at the next day the accumulation of zeatin riboside and zeatin was registered in the shoots of treated plants. The increase in cytokinin content promoted plant growth (the increased leaf length and width and a faster accumulation of plant fresh and dry weight). Plant treatment with a substance obtained from microorganisms incapable to synthesize hormones resulted in the insignificant growth stimulation. Plant treatment with a substance obtained from cytokinin-producing microorganisms increased leaf chlorophyll content; in this case, the level of chlorophylls was comparable to that observed in the plants treated with a synthetic cytokinin benzyladenine. The role of cytokinins of microbial origin as a factor providing for growth-stimulating effect of bacteria on plants is discussed.     

Grain or coleoptile tip as the source of IAA in cereal shoots?

Abstract. Current opinion is divided on whether the free IAA in cereal shoots is derived from the coleoptile tip or from the grain. To date these hypotheses have been considered to be mutually exclusive. In this paper the idea is advanced that both sources may function in the supply of IAA in the shoot. Critical review of the literature shows that such an explanation best fits all the available evidence. It is argued that the relative importance of each source may vary between species of the Gramineae, with the supply of IAA in Avena shoots being derived predominantly from the grain, whereas in Zea the coleoptile tip is the more important source.     

Effect of light on the organogenic ability of garlic roots using a one-step in vitro system

A simple and efficient garlic in vitro shoot regeneration protocol has been developed. This system uses axenic root tips cultivated from the beginning in the presence of light and does not require any change or refreshing of the original medium during the entire process. The application of light from the beginning of the culture process did not affect the callus formation rate but did significantly improve the explant regeneration ability. In a 2-month period it was possible to obtain up to 250 shoots per gram of callus.Abbreviations BAP: 6-Benzylaminopurine - 2,4-D: 2,4-Dichlorophenoxyacetic acid - NAA: -Naphthaleneacetic acid Communicated by L. Peña     

Role and function of cytokinin oxidase in plants

Cytokinin oxidase (CK oxidase) is widely distributed in plants and is the only enzyme that has been shown unequivocally to catalyze the catabolism of specific cytokinins (CKs) to inactive products that lack the N⁶-unsaturated side chain. Thus, the enzyme is thought to play a major role in controlling the level or species of CKs in plant tissues. However, despite its discovery more than 25 years ago, little attention has been given to the elucidation of its role and function in plant growth and development. This review seeks to bring in to context the current state of knowledge regarding the biochemical and molecular properties, regulation in undifferentiated and differentiated tissues, and recent results from studies using transgenic plants in an attempt to provide a more comprehensive understanding of the physiological significance of the enzyme in plants. Notwithstanding species, tissue and other specific differences, in general, CK oxidase appears to contribute to CK homeostasis in plants. However, complete clarity as to its function awaits purification of the protein to homogeneity and the ultimate development of requisite molecular probes.     

Growth and biochemical responses of jojoba (Simmondsia chinensis (Link) Schneid) explants cultured under mannitol-simulated drought stress in vitro

Jojoba explants were cultured under four levels of mannitol (control plus 50, 100, 250 and 500 mM mannitol)-induced osmotic stress during the proliferation stage in vitro. Explants grown under control condition exhibited the highest growth, while the more severe the stress was, the lower was the growth of explants. Electrolyte leakage, lipid peroxidation, H₂O₂ content and proline concentration were highest and relative water content lowest under the highest level of osmotic stress. Concentration of phenolic compounds (total phenolic compounds, o-diphenols and flavanols, as well as protocatechuic, vanillic, p-coumaric and ferulic acids and rutin), putrescine and total polyamines decreased with increasing stress level. Mannitol, glucose and pinitol concentrations increased, whereas that of inositol decreased with increasing stress level. Explants were transferred to the rooting stage, separately per stress treatment. Explants grown under stress conditions during the proliferation stage exhibited lower rooting percentage than controls, as the stress became more severe, the lower was the rooting response. Jojoba tolerated osmotic stress to some extent (till 100–250 mM mannitol), exhibiting sufficient growth rate and good rooting response as well as low oxidative damage (based on electrolyte leakage and lipid peroxidation indices).     

In vitro PPFD and media composition affect both in and ex vitro performance of Alstroemeria Butterfly-hybrids

The objective was to reduce in vitro production costs while retaining or improving plant quality, in particular the suitability for pot plant production. Plants were grown at photosynthetic photon flux densities (PPFD) of 0–40 μmol m^-2 s^-1 and sucrose concentrations of 3–7% during the multiplication phase and the effects of sucrose, BA, and NAA during root formation were investigated. Ex vitro growth were tested in both experiments. A small reduction in the rhizome multiplication rate was found with increasing PPFD and sucrose concentration. Increasing sucrose concentration reduced the number of aerial shoots. Aerial shoots were etiolated when cultured in darkness and their number increased with increasing PPFD at 3% sucrose, whereas PPFD did not affect the number of aerial shoots at 5 or 7% sucrose. During the multiplication phase a synergistic promoting effect of PPFD and sucrose was observed on root formation. Root formation after transfer to rooting medium was affected by sucrose and PPFD during the multiplication phase. PPFD did not influence root formation after propagation on 7% sucrose, whereas on 3 or 5 % sucrose root formation was gradually inhibited when PPFD was decreased below 17 μmol m^-2 s^-1. The formation of thick roots was promoted by propagation in light, and not influenced by sucrose concentration. Ex vitro growth was not affected by in vitro conditions, except for 7% sucrose during the multiplication phase that reduced flowering. Root formation on rooting medium was reduced by BA and promoted both by NAA and high levels of sucrose. The root inhibiting effect of BA could not completely be overcome by simultaneous application of NAA and high sucrose concentrations. Thick roots were only produced in the presence of NAA, and not affected by sucrose treatment. Ex vitro flowering was negatively influenced by the presence of BA during root formation and by high levels of sucrose if BA was absent in the rooting medium. High sucrose levels and NAA could partially compensate for the negative effect of BA on flowering. This revised version was published online in June 2006 with corrections to the Cover Date.