Does light spectral quality affect survival and regeneration of potato (Solanum tuberosum L.) shoot tips after cryopreservation?

Cryopreservation, the storage of germplasm at ultra-low temperature is the most reliable tool for long-term preservation of plant genetic resources. Cryopreservation techniques are widely applied but the effect of light spectra on plant recovery after cryopreservation is largely unknown. Therefore, we investigated the effect of different light spectral qualities on survival and regeneration of shoot tips of potato (Solanum tuberosum L.) cultivars Agrie Dzeltenie, Maret, Bintje, Désirée and Anti cryopreserved by the DMSO-droplet method. Prior to cryopreservation, the plants were stored under cool white fluorescent light (CW). Post-cryopreservation, the plants were allowed to regenerate under six different light qualities: CW, warm white light (HQI), blue LEDs (B), red LEDs (R), red with 10 % of blue (RB) and RBF - red with 10 % of blue with addition of 20 % of far-red LEDs. The light spectral quality had a significant effect on the survival and regeneration of potato shoot tips after cryopreservation. The combination of red light with 10 % of blue (RB) doubled the regeneration percentage of all cultivars, whereas red light (R) was not suitable for regeneration after cryopreservation. Specifically, the regeneration percentages were increased in RB compared to CW from 25.5 to 52.6 % for ‘Agrie Dzeltenie’, 25.0–43.6 % for ‘Maret’, 8.1–26.1 % for ‘Bintje’, 0.0–17.1 % for ‘Anti’ and 18.2–36.6 % for ‘Désirée‘. Therefore, the modification of light spectra during the recovery phase is a promising tool for increasing the regeneration of potato shoot tips after cryopreservation.     

Effects of drought stress on seed germination of ornamental sunflowers

The response to drought stress on germination was investigated on three hybrids of ornamental sunflower, ‘Hadar’, ‘Pazit’, and ‘Zohar’. Different levels of water potential [Ψ: 0.0 (control), ?0.15, ?0.30, ?0.45, ?0.60, ?0.75, and ?0.90 MPa] were adopted using polyethylene glycol-6000 (PEG6000) at four germination temperatures (15, 20, 25, and 30 °C). Final germination percentage, mean germination time, germination index, germination rate index, and germination stress tolerance index were used to evaluate the genotype response to PEG-induced water stress. Shoot and root length and fresh and dry weight were measured on seeds germinated at 20 °C under the different levels of water potentials. During germination, the three ornamental sunflowers showed to be more sensitive to suboptimal temperature than to supraoptimal. Decreasing water potential of imbibition solution progressively inhibited and delayed seed germination. Among cultivars, ‘Hadar’ and ‘Pazit’ performed better at temperature lower than 30 °C. ‘Zohar’ showed a lower sensitivity to PEG-induced water stress at all temperature conditions. Water stress during seed germination depressed the following seedling growth under favourable conditions. As a result, shoot and root length and fresh and dry weight was significantly lower in seedlings from seed germinated at ψ ≤ 0.45 MPa.     

Assessment of effective pollination period in olive (Olea europaea) by means of fluorescence microscopy and plant response to sequential pollinations: limitations and drawbacks of current methodologies

Key message

The olive tree response to sequential pollinations is a more accurate procedure in the assessment of effective pollination period and flower life span than procedure based on the calculation of the components of the pollen-pistil interaction.

Abstract

Effective pollination period (EPP) was defined by Williams (1965) as the period during which pollination can result in fruit set. EPP was proposed to be the time the ovules remain fertile minus the time needed for a pollen tube to reach the ovule, providing that this value does not exceed the length of stigma receptivity. The duration of the EPP in fruit trees can be estimated by the microscopic analyses of these three components, but also by the plant response to sequential pollinations in term of fruit set. Here, we compare the estimation of EPP duration under cross-pollination in the main olive cultivars (Olea europaea L.) grown for oil production in Croatia (‘Lastovka’, ‘Leccino’, and ‘Levantinka’) based on these two procedures to discuss the advantages and disadvantages of each estimation and propose the best methodology to assess EPP duration and the capacity of flowers of different age to set fruit. Our comparison highlights that the different procedures provide a contrasting duration of EPP in all olive cultivars here analyzed. EPP based on initial fruit set and final fruit set was between 2 and 3 days, while that based on the microscopic study of EPP components was between 5 and 12 days, depending on the cultivar. We defend that EPP based on fruit set is more accurate and more relevant for growers, and believe that the longer duration of EPP when the estimation is based on the study of the components could be due to wrong assessments of the duration of particular components. In this sense, pollen germination may be a better parameter than pollen adhesion to estimate stigma receptivity. Ovule longevity assessment based on the fluorescence emitted could be an overestimation of the actual ovule readiness for fertilization. Style suitability, not considered in Williams’ equation, plays an important role supporting pollen tube growth toward the ovules and might be also relevant in determining EPP duration.     

Cloning and expression of a tau class glutathione S-transferase (ZjGSTU1) from Chinese jujube in response to phytoplasma infection

Phytoplasmas are associated with diseases of several hundred plant species. Jujube witches’ broom disease (JWB) is a destructive phytoplasma disease in Chinese jujube (Ziziphus jujuba Mill.). Ziziphus jujuba Mill. ‘J5’, an excellent strain with extremely high resistance to JWB, was selected by us. In our previous study, a GST (EC 2.5.1.18) fragment was sequenced from suppression subtractive hybridization library of ‘J5’ under JWB phytoplasma stress. Based on this result, a GST gene (ZjGSTU1, HM345954) was first isolated from jujube by homology cloning and RACE. ZjGSTU1 contains a complete open reading frame of 702 bp encoding a protein of 233 amino acid residues. Sequence alignment showed that ZjGSTU1 shared a typical conserved structure and high identity with tau GSTs from other plant species. Relative RT-PCR demonstrated that the expression of ZjGSTU1 mRNA in jujube leaves and branches could be triggered by JWB phytoplasma. Moreover, the expression of ZjGSTU1 mRNA in resistant strain ‘J5’ increased sooner and higher than that in sensitive strain ‘J9’ under JWB phytoplasma stress (‘J5’ and ‘J9’ are two strains from the same cultivar). Western blotting analyses showed that the expressions of ZjGSTU1 in ‘J5’ and ‘J9’ were dramatically up-regulated under JWB phytoplasma stress and its expression in ‘J5’ was also higher than that in ‘J9’ at protein level. Collectively, this paper highlights that ZjGSTU1 gene is responsive to phytoplasma infection. The possible roles of this gene were discussed in terms of regulatory process in resistance to phytoplasma infection.     

Influence of frost damage on the sugars and sugar alcohol composition in quince (Cydonia oblonga Mill.) floral nectar

Cold stress adversely affects growth and productivity, and triggers a series of morphological, physiological, biochemical and molecular changes in plants. Since sugars are present in all floral nectars in greater amounts than any other constituent, the aim of this study was to examine how frost exposure changes sugar metabolism and how it affects on the content of sugar components in the nectar of quince. Three quince cultivars (‘Vranjska’, ‘Triumph’ and ‘Leskova?ka’) were investigated in this study. The contents of sugars (glucose, fructose, sucrose, trehalose, maltose, isomaltose, rhamnose, arabinose, ribose, melezitose, raffinose, and panose) and sugar alcohols (sorbitol, erythritol, mannitol and galactitol) were analyzed by high performance anion exchange chromatography (HPAEC) with amperometric detection. The results showed that after late spring frosts and irreversible damage of flower parts, the nectar of the three quince cultivars contained elevated levels of fructose, trehalose, arabinose, ribose, rhamnose, raffinose, galactitol and mannitol, indicating an impairment of central carbohydrate metabolism. The ratios between individual sugars, such as the glucose/fructose ratio, were changed in the nectar of damaged flowers in all three quince cultivars. The examined cultivars showed similar sugar response to cold stress. The only exception was ‘Leskova?ka’ for the glucose and melezitose pathway, which means that composition of those two sugars changed significantly according to the genotype. The larger are the carbohydrates reserves in different parts of a fruit tree, the higher is the tolerance to any form of frost damage, the results of this study could help in the understanding of how different quince cultivars react to this kind of stress and how they modulate their sugar metabolism.     

Comparative electrophoretical analysis of plasmid-like mitochondrial DNAs in Vicia faba and in some other legumes

Minicircular DNAs found in mitochondria of 6-d-old etiolated seedlings of Vicia faba L. were also present in mitochondria isolated from cell suspension cultures and from green leaves of this plant. These results support the suggestion that the plasmid-like molecules found in mitochondria of V. faba are an essential component of the mitochondrial genome. The minicircular DNAs were, apparently, peculiar for the species V. faba since they were found in all three cultivars of this species which were studied. The distribution pattern of plasmid-like DNAs in Vicia villosa L. was completely different and mitochondria from Medicago sativa L. also contained specific minicircular DNAs. Thus, minicircular DNAs are typical for the mitochondrial genomes of several legumes and different plant species have their specific mitochondrial plasmid-like DNAs     

Resistance of Citrus and Related Genera to Diaphorina citri Kuwayama (Hemiptera: Liviidae)

The present study was developed to evaluate the resistance of the following genotypes of Citrus and related genera to this pest: ‘Pera,’ ‘Natal’, and ‘Washington Navel’ oranges (Citrus sinensis), ‘Marsh Seedless’ grapefruit (Citrus paradisi), hardy orange ‘Rubidoux’ (Poncirus trifoliata), kumquat (Fortunella margarita Swingle), citrumelo ‘Swingle’ (C. paradisi x P. trifoliata), and citrange ‘Troyer’ (P. trifoliata x C. sinensis). The experiments were performed in greenhouses with plants grafted onto ‘Rangpur’ lime (Citrus limonia) and placed individually in voile cages. The preference for oviposition in a no-choice test, and the effect of genotype were evaluated. The egg-adult cycle was monitored to determine the effect of genotype on the biology of the insect. Poncirus ‘Rubidoux’ was the least preferred genotype for oviposition; reduced number of eggs was also found to occur on citrange ‘Troyer’, and ‘Marsh Seedless’ was the genotype with the most eggs. No significant variation in the duration of the embryonic period was observed; however, a difference in the viability of eggs was found, with the lowest egg viabilities on ‘Swingle.’ Kumquat and ‘Marsh Seedless’ genotypes were correlated with increased durations of the nymphal phase, however, there was no difference in the survival of this phase. Fecundity of females on ‘Troyer’, ‘Swingle’, and kumquat was reduced. Considering all of the evaluated parameters, it was concluded that cultivars of sweet orange are the most susceptible genotypes to Diaphorina citri. Regarding oviposition, P. trifoliata ‘Rubidoux’ showed resistance of the antixenosis type.     

Changes in the activities of catalase,peroxidase, and polyphenol oxidase in apple buds during bud break induced by thidiazuron

The breaking of dormancy in apple buds (Malus domestica Borkh cv. York Imperial) by thidiazuron (N-phenyl-N′-1,2,3,-thidiazol-5-ylurea) was investigated in relation to catalase, peroxidase, and polyphenol oxidase activities and their isoenzyme patterns. The activity and number of isoenzymic components of catalase increased progressively during bud break, then decreased after buds started to grow. Peroxidase activity was highest during dormancy and declined during bud swell, increased at bud break, and decreased after bud expansion. Several isoperoxidases were observed in gel electrophoresis. Similar patterns were found at different growth stages of apple buds except for one peroxidase isoenzyme, P3, which disappeared 12 days after thidiazuron treatment. There was an inverse relationship between the activities of polyphenol oxidase and peroxidase during the development of apple buds. Apple buds have a very similar polyphenol oxidase isoenzyme pattern throughout bud development. However, the appearance and disappearance of minor isoenzymes were also observed. Phloridzin, rutin, p-coumaric, epicatechin, naringin, chlorogenic acid, and catechol were found in apple buds. Among them, phloridzin, rutin, and p-coumaric were the dominant phenolic compounds. Dormant buds contained a high amount of phenolic substances which decreased after bud break (4 days after thidiazuron treatment) then increased until the start of bud expansion. Phenolic compounds are found to be potent modifiers of catalase, peroxidase, and polyphenol oxidase activity, as both inhibitors and stimulators in apple buds.     

The separation and comparison of albumin complexes of seed proteins in three cultivars ofPhaseolus vulgaris

Complexes of water soluble proteins (albumins) were investigated in three cultivarsof Phaseolus vulgaris, viz: Yeltruská Saxa, Vainica Saavegra B, and Krupnaya sakharnaya. The first two cultivars exhibit haemagglutinating activity against rabbit erythrocytes, but have different elution profiles on Sephadex G-100. Their individual peaks have a different subunit composition, as revealed by SDS gel electrophoresis, as well as a different immunoelectrophoretic pattern, although proteins I and II of the specificity Veltruská Saxa are present in both cultivars. The cultivar Krupnaya sakharnaya expressively differs from the preceding lectin cultivars; it has no erythroagglutinating activity, its albumin complex has a high-molecular component, absent in the preceding ones, and has no lectin peak in the region of molecular mass of 100 000 to 200 000. Immunoelectrophoresis gave no evidence of protein I and II of the specificity Veltruská Saxa.     

Tissue and protoplast culture studies in Lycopersicon esculentum miller var. flammatum lehm. cv. ‚Bonner Beste‘ and its mutant chloronerva

Methods of shoot and plant development from cotyledon explants of the tomato cultivar ‘Bonner Beste’ and its mutant chloronerva are described. Cotyledon mesophyll protoplasts of the mutant chloronerva have been isolated and cultured, and induced to regenerate shoots and plants. By grafting regenerated shoots of the chloronerva mutant on rootstocks of the cultivar ‘Bonner Beste’ plants were obtained producing fruits and seeds. Plants derived from these seeds show all the typical characteristics of the chloronerva mutant.     

The synthesis of short- and long-chain acylarnitine by etio-chloroplasts of greening barley leaves

Etio-chloroplasts of barley, purified on sucrose density gradients were shown to possess carnitine long-chain acyltransferase (carnitine palmitoyltransferase, EC 2.3.1.21) activity and carnitine short-chain acyltransferase (carnitine acetyltransferase EC 2.3.1.7) activity. These enzymes may play a role in the transport of acyl groups as acylcarnitines through the membrane barrier of barley etio-chloroplasts and also ‘or alternatively’ may spare CoA by transferring short- and long-chain acyl groups from short-and long-chain acyl CoA to carnitine.     

Genome-wide single nucleotide polymorphisms and insertion–deletions of Oryza sativa L. subsp. japonica cultivars grown near the northern limit of rice cultivation

Single nucleotide polymorphisms (SNPs) and insertions–deletions (InDels) are valuable molecular markers for molecular breeding among genetically closely related cultivars. Rice (Oryza sativa L. subsp. japonica) cultivars grown in Hokkaido (45–42°N), the northernmost region of rice paddy cultivation in Japan, have been bred for over 100 years for adaptation to low summer temperatures together with high yield and good eating quality. In this study, for 10 closely related rice cultivars released in Hokkaido and cultivar Koshihikari, we identified genome-wide SNPs and InDels by next-generation sequencing. More than 29 million reads from the Hokkaido cultivars, each 101 nucleotides long, were uniquely mapped to the Nipponbare reference genome. The average of the total nucleotide length of all uniquely mapped reads corresponded to 10.9 times (3,978 Mb with genome coverage of 90.7 %) the Nipponbare reference genome. An average of 99,955 putative SNPs (1.8 times the number in Koshihikari) and 14,617 putative InDels (also 1.8 times the number in Koshihikari) were detected in Hokkaido cultivars relative to the Nipponbare genome, which enabled analyses of the inheritance of pedigree haplotypes of four cultivars, SNPs and InDels among closely related Hokkaido cultivars, and haplotype blocks unique to Hokkaido cultivars. The comprehensive SNP and InDel data provide DNA marker resources and will facilitate quantitative trait locus analysis of biparental mapping of very closely related Hokkaido cultivars. Furthermore, the haplotype blocks unique to Hokkaido cultivars represent ideal genetic regions for improvement of cultivars to be grown near the northern and southern limits of rice cultivation.     

Aspergillus insulicola sp. nov.

A strain of Aspergillus sp. is described and proposed as a new species under the name ‘Aspergillus insulicola sp. nov.’ Montemayor & Santiago, 1973. This strain was isolated from soil samples taken in ‘Aves Island’ during a scientific expedition. — Aves Island, situated at 15°, 40′, 42″ N and 63°, 36′, 47″ W, about 665 Km of the coast of Venezuela, has very special ecological conditions. Due to its smallness: 550 m long and 40 to 120 m across and to its low profile only 3 m over sea level, it is swept by the sea during the periodical storms and hurricanes in the area. It has thus a very interesting fauna and flora. We took a series of soil samples to study its mycological flora. Forty samples were inoculated by dilution method. In this first paper a species is described and proposed as a new species because of its macroscopic and microscopic characteristics, as well as by its biological properties, under the name ‘Aspergillus insulicola sp. nov.’. In its study we have tried to follow as closely as possible the methods recommended by Kennet B. Raper & Dorothy Fenell, world authorities on the genera Aspergillus and Penicillium. The strain is being kept in USB under the number T1, and has been sent to ATCC & CBSC to be incorporated in their collections.