Demeter Lásztity

Effect of short-term and long-term low temperature stress on polyamine biosynthesis in wheat genotypes with varying degrees of frost tolerance

Summary Two series of experiments were carried out to examine the short- and long-tetm effects of low temperature on polyamine biosynthesis in wheat. In the first series, studies were made on the polyamine accumulation in the leaves, crowns and roots of winter wheat varieties with varying degrees of frost toletance subjected to short-term low temperature stress (6h, -2 °C). A marked accumulation of Put was observed. Agm accumulation was also examined and found comparable to that of Put. This suggests that Agm, which is an intermediate product of Put synthesis only in higher plants, may play an important role during short-term cold treatment. The second series of experiments was aimed at discovering the effect of wheat chromosomes 5A and 7A, which contain major genes responsible for frost resistance, on the polyamine synthesis taking place in various parts of the seedlings during long periods of cold treatment, and especially on the alternative metabolic pathway present only in higher plants.

S-methylmethionine reduces cell membrane damage in higher plants exposed to low-temperature stress

S-methylmethionine (SMM), an important intermediate compound in the sulphur metabolism, can be found in various quantities in majority of plants. The experiments were designed to determine the extent to which SMM is able to preserve cell membrane integrity or reduce the degree of membrane damage in the course of low-temperature stress. By measuring electrolyte leakage (EL), it was proved that SMM treatment reduced cell membrane damage, and thus EL, during low-temperature stress in both the leaves and roots of peas, maize, soy beans and eight winter wheat varieties with different levels of frost resistance. Investigations on the interaction between SMM and polyamine biosynthesis revealed that SMM increased the quantities of agmatine (Agm) and putrescine (Put) as well as that of spermidine (Spd), while it had no effect on the quantity of spermine (Spn). Using a specific inhibitor, methylglyoxal-bis-guanyl hydrazone (MGBG), it was proved that the polyamine metabolic pathway starting from methionine played no role in the synthesis of Spd or Spn, so there must be an alternative pathway for the synthesis of SMM-induced polyamines.

Tests of species concepts of the small, white, European group of Tuber spp. based on morphology and rDNA ITS sequences with special reference to Tuber rapaeodorum

Several taxonomic problems arise in the group of small, white European truffles, probably due to the over-emphasized significance of certain morphological features of ascomata. The distinction between Tuber rapaeodorum Tul. & C. Tul. and Tuber borchii Vittad. and Tuber puberulum Berk. & Broome has not been accepted in several recent studies. Furthermore, the existence of T. rapaeodorum been questioned in some recent synopses of the genus. We conducted microscopic and ITS sequence investigations of 31 herbarium specimens. Using morphological features such as peridium structure, form and size of spores and dermatocystidia and spore numbers per ascus, we could distinguish T. borchii, T. foetidum Vittad., T. maculatum Vittad., Tuber puberulum, and T. rapaeodorum. Analysis of whole ITS sequences showed sharp differences among the morphologically separated groups. Neighbour-joining and parsimony methods produced highly supported branches and confirmed the identity of these species.

Changes in the Content of Modified Nucleotides in Wheat rRNA during Greening

The modified nucleotide content of the ribosomal RNAs in wheat is greatly influenced by light. The rRNAs of etiolated seedlings contain far fewer modified derivatives. The modified nucleotide composition characteristic of green plants develops gradually as a result of irradiation. In the course of the experiments changes in the state of modification of 5.8S and 18S rRNAs were examined during the greening of etiolated wheat seedlings. Three types of minor nucleotides, O2′-methyladenosine, O2′-methylguanosine and pseudouridine were found in the 5.8S rRNA of green wheat leaves, none of which was detected in etiolated wheat. The minor nucleotides appeared in the 5.8S rRNA only after 48 h irradiation. The sequences of 5.8S rDNA, TTS1, ITS2 and 18S rDNA were also determined and the presence of the hyper-modified nucleotide 1-methyl-3-(α-amino-α-carboxypropyl)-pseudouridine was detected in green wheat 18S rRNA. This minor component was not demonstrable in etiolated wheat 18S rRNA, but appeared after irradiation for 48 h.

Are either SOD and catalase or the polyamines involved in the paraquat resistance of Conyza canadensis

Paraquat/atrazine coresistant (PqAR) and paraquat resistant (PqR) horseweed (Conyza canadensis L. Cronq.) plants showed - in the first hour after 0.5 mM paraquat spraying - a decreased catalase activity followed by a slight increase. However, the enzyme activity remained always below the initial value Sensitive plants showed a significant increase of catalase activity in the first 4 hour after spraying The transient character of paraquat inhibition, the recovery of photosynthetic activity of the PqAR Conyza plants (characterized by variable fluorescence) after spraying remained unaffected by the superoxide dismutase (SOD) inhibitor, diethyldithiocarbamate This indicates that SOD is not involved in the resistance mechanism. Untreated resistant biotypes showed about 2.5 times higher total polyamine and putrescine level than the sensitive one. 100 μM of exogenously added putrescine was observed as having a protecting effect against paraquat in floated leaves of the sensitive biotype only. The resistant leaves were unaffected probably on account of their higher endogenous polyamine level. It is concluded that polyamines may play a role in the paraquat resistance of Coniza canadensis.

Paraquat resistance of weeds--the case of Conyza canadensis (L.) Cronq.

The paper gives an overview of literature on paraquat resistance of weeds and the proposed mechanism of resistance. New results we achieved on horseweed ( Conyza canadensis /L./, Cronq.) are discussed in detail. It was demonstrated that there is no significant constitutive difference related to the paraquat resistance between untreated susceptible and paraquat-resistant horseweed plants. The lower sensitivity of flowering resistant plants may be due to the fact that paraquat content in treated leaves of flowering resistant plants was only 25% as compared to those measured at rosette stage. Our results confirm that paraquat resistance is not based on elevated level and activity of antioxidant enzyme system. The hypothesized role of polyamines in the resistance mechanisms can be excluded. The higher putrescine and total polyamine content of paraquat treated resistant leaves can rather be regarded as a general stress response, than as a symptom of paraquat resistance. A paraquat-inducible protein is supposed to play a role in the resistance, which presumably functions by binding paraquat to an inactivating site and/ or by carrying paraquat to metabolically inactive cell compartment (vacuole, cell wall). From model experiments it is concluded that paraquat and diquat preferentially form hydrophylic interactions with proteins containing a higher amount of lysine and glutamic acid. Consequently, the reason for paraquat resistance in horseweed is probably a hydrophylic interaction of paraquat with a protein, leading to inactivation of paraquat through forming a conjugate and/or sequestration into the vacuole or the cell wall.

Intraspecific invariability of the internal transcribed spacer region of rDNA of the truffle Terfezia terfezioides in Europe

ITS regions (internal transcribed spacers—ITS1 andITS2—with the 5.8S gene of the nuclear rDNA) of 25 fruit body samples ofTerfezia terfezioides, originating from Hungary and Italy, were compared. The amplification and sequencing of the ITS region was successful with both theITS1-ITS4 andITSIF-ITS4 primer pairs. No differences of the restriction fragment length polymorphism profiles were detected among 19 samples collected in one place at the same time. The sequences of the ITS region of 9 samples collected in different localities were highly invariable, differing in only two bases. Thus the intraspecific homogeneity of the ITS region seems to be an important species-specific characteristic ofT. terfezioides in contrast to otherTerfezia species. As the samples of the species were collected from different and distant localities of Europe, the ITS sequence ofT. terfezioides can be considered a very conservative, reliable molecular marker of the fungus. *** DIRECT SUPPORT *** A00EN076 00008

Changes in the content of modified nucleotides of total transfer RNA of wheat seedlings during greening.

The contents in minor nucleotides of total transfer RNA (tRNA) of etiolated and light-grown wheat (Triticum aestivum L.) seedlings and of seedlings illuminated for 24 or 48 h were examined. The total tRNA of seedlings illuminated 24 h contained more, and that from seedlings illuminated 48 h still more modified nucleotides than that from etiolated ones. Thus, the appearance of the characteristic minor nucleotides of tRNA of light-grown wheat seedlings needs a rather long greening period, of at least 48 h.

Effect of S-methylmethionine on the photosynthesis in maize at different chilling temperatures

The effect of the natural compound S-methylmethionine (SMM) on the functioning of the photosynthetic apparatus, the efficiency of photosynthesis and the synthesis of stress-induced phenoloids and anthocyanins involved in defence was investigated in young maize plants exposed to moderate and severe chilling stress. Damage to PSII was observed as a reduction in the value of variable fluorescence (Fv/Fm) which could be detected even after few hours of mild chilling stress. At temperatures below 10°C, the reduction in Fv/Fm was more pronounced. Changes in the value of net photosynthesis exhibited a similar tendency. SMM has a moderating effect on this reduction and its protective effect was more pronounced under long-lasting chilling stress and at the lowest temperatures. Monitoring of fluorescence intensities and ratios correlated with the levels of stress defence compounds. The fluorescence intensities were found to increase over the course of chilling stress in response to SMM, with the highest values being recorded in plants exposed to the longest period of stress. A similar tendency was observed for the quantity of anthocyanins. The results confirm the complex role of SMM, which is manifested both in preserving the ability of the photosynthetic apparatus to function and in stimulating the synthesis of metabolites involved in stress defence.

Effect of long periods of low temperature exposure on protein synthesis activity in wheat seedlings

Abstract Long periods of low temperature exposure induce complex changes in the metabolism of nucleic acids and protein molecules in plants: i.e. new proteins; new tRNA isoacceptors and new mRNAs appear with altered minor nucleotide contents, implying that the components of the protein synthesising system change during cold treatment. To study the effect of changes in the RNA pool on the intensity of protein synthesis, different homologous and heterologous cell-free protein synthesising systems were constructed with polysome fractions and tRNAs isolated from non-treated wheat seedlings and from seedlings cold treated for a long period. The homologous cell-free protein synthesising systems contained polysome fractions from non-treated samples of the wheat cultivar Martonvasari 15 and from samples treated for 1, 5 or 7 weeks together with their own tRNA. Heterologous systems were constructed from the tRNA fractions of cold-treated seedlings with S 23 fractions of non-treated ones and vice versa. Cell-free protein synthesis was carried out at 4 and 30°C. The results demonstrate that independently of the length of the cold period the intensity of protein synthesis in homologous cold-treated systems at 4°C was as high as the intensity of homologous non-treated systems at 30°C. Combinations of cold-treated S 23 fractions with cold-treated tRNAs were about 30% more effective than cold-treated S 23 fractions with non-treated tRNAs at 4°C, while combinations of cold-treated tRNAs with non-treated S 23 fractions resulted in only a slight decrease in activity at 30°C. It can thus be concluded that long-term cold exposure leads to changes in the protein synthesising system, resulting in optimal synthesising capacity under the altered conditions.