Increase in urinary markers during the acute phase reflects the degree of chronic tubulointerstitial injury after ischemia-reperfusion renal injury

Context: Acute kidney injury (AKI) could lead to progressive chronic kidney disease (CKD). Objectives: To demonstrate that urinary markers in AKI are associated with the degree of persistent renal injury. Material and methods: Human L-FABP chromosomal transgenic (T_g) mice were subjected to ischemia-reperfusion (I/R) clamping renal pedicle for 20?min or 30?min. Kidneys were obtained at one and 40 days after I/R. Results: Urinary L-FABP, NGAL, Kim-1 and albumin levels increased during the acute phase and were significantly correlated with the degree of tubulointerstitial fibrosis during the chronic phase. Discussion and conclusion: These markers could detect higher risk of progression to CKD.     

Urinary mRNA expression of CCN2/CCN3 as a noninvasive marker for monitoring glomerular structure changes in nondiabetic chronic kidney disease

Background: We investigated whether urinary mRNA of connective tissue growth factor (CCN2) and nephroblastoma overexpressed gene (CCN3) can provide clinical insight into the management of patients with nondiabetic CKD.

Methods: Urinary mRNA expression of CCN2 and CCN3 were measured by Real-time PCR in 35 CKD patients and 12 controls.

Results: Urinary mRNA of CCN2 and CCN3 were distinctively greater in CKDs than healthy controls. Urinary CCN3/CCN2 mRNA ratio correlated to the degree of glomerular histological changes in those who received renal biopsy.

Conclusion: Urinary CCN3/CCN2 mRNA ratio may be a useful noninvasive biomarker for evaluating patients with nondiabetic CKD prior to renal biopsy.     

Flower Bud Culture of Shallot (Allium cepa L. Aggregatum group) with Cytogenetic Analysis of Resulting Gynogenic Plants and Somaclones

Unpollinated flower culture was applied for induction of gynogenesis and somatic organogenesis in three shallot strains, ‘Dili-white’, ‘Yogya’ and ‘Dili-red’, from Indonesia. Chromosome surveys were performed on the plants obtained. From a total of 6,812 flowers, 89 plantlets were obtained by gynogenesis, of which 10 could be acclimated. Most of the plantlets were induced from ‘Dili-white’. Of the gynogenetic plants examined, two were haploid (2n = 8), four were naturally doubled haploid (2n = 16) and the remaining four were mixoploid (2n = 8, 16 or 2n = 16, 32, 64). ‘Dili-red’ showed the highest frequency of somatic organogenesis. Fifty-nine directly regenerated plants and 293 callus-derived plants were obtained from somatic organogenesis from the cultured flowers. Based on the chromosome number, frequencies of somaclonal variation were high both in the directly regenerated plants and the callus-derived plants. The frequency of tetraploid plants (2n = 32) in the former (50%) was higher than in the latter (33%). From these results we conclude that unpollinated flower culture is an effective method for chromosome doubling simultaneously with haploid induction in shallot.     

Karyotype with 210 chromosomes in guaraná (<Emphasis Type="Italic">Paullinia cupana</Emphasis> ‘Sorbilis’)

The genus Paullinia includes the economically important P. cupana, known as guaraná in Brazil and more recently in the world market. Native Americans of the Maué and Andirá tribes cultivated P. cupana ‘Sorbilis’ in central Amazon, and the Barés cultivated the ‘Typica’ variety in the upper Negro River (Brazil). Cytological studies in the Sapindaceae family have concentrated on the diversity in number (from 2n = 14 to 96) and size of the chromosomes. In Paullinia, seven species have been karyotyped and all show 2n = 24. Meristem maceration, cellular dissociation and air-drying techniques were used for cytogenetic preparations and DNA content was determined by flow cytometry. Chromosome characterization and DNA content of Paullinia cupana Kunth ‘Sorbilis’ (Mart.) Ducke (Sapindaceae) were studied. The high chromosome number (2n = 210) fall into two cytomorphological groups: (a) a metacentric and submetacentric group showing 25 sets of three pairs of chromosomes (2–76); (b) a group containing only acrocentric showing 12 sets of two pairs of chromosomes (82–105), a homologous submetacentric pair (1) and an acrocentric pair (81). Mean nuclear DNA content of guaraná was 2C = 22.8 pg. A karyogram was set up showing a high chromosome number complement.     

Small plot mating disruption trials for tufted apple bud moth,Platynota idaeusalis, in Pennsylvania apple orchards

Mating disruption treatments for the tufted apple bud moth (TABM),Platynota idaeusalis (Walker), were tested in small plot trials in apple orchards in Pennsylvania. Treatments were evaluated by fruit injury and by capture of male TABM in traps baited with synthetic pheromone sources or virgin females. The TABM pheromone is a two component isomeric blend ofE-11-tetradecen-1-ol (E11-14:OH) andE-11-tetradecenyl acetate (E11-14:Ac). A 50∶50 ratio of these two components was used in standard monitoring septa and in mating disruption treatments released from either hollow fibers (‘fiber’) or PVC tubes (‘PVC’). Other pheromone blends tested included a 90∶10 ratio of E11-14: Ac and E11-14:OH (‘EAc’) and its reverse (‘EOH’), mixture ‘EAc’ with 30% of the Z-isomers (‘low AEc’), and a blend similar to the preceding with 2% Z9-12:Ac (‘generic’). These other blends were released from multi tube tape (‘tape’) or Shin-Etsu type rope (‘rope’) dispensers. Seasonal dispenser release rate in mg ha⁻¹ h⁻¹ was ca. 30 for the ‘rope’ dispensers, 14 for ‘PVC’ and 6 for ‘fiber’. ‘EAc-tape’ and ‘EOH-tape’ were equally effective in reducing catches of males in traps baited with synthetic lures and in traps baited with virgin females. Both treatments also reduced fruit injury. ‘EAc-rope’, ‘fiber’ and ‘PVC’ also were generally effective; whereas, the ‘low EAc’ and ‘generic’ treatments reduced trap capture less than 90% and did not reduce fruit injury. Dispenser density was positively correlated with reduction in trap capture for the ‘low EAc-rope’ and ‘genericrope’ treatments. Traps loaded with ‘fiber’ dispensers captured more male TABM than the other treatments in non-pheromone permeated environments. Trap capture of other tortricids was reduced in pheromone treatments. ‘EAc-rope’ and the ‘TABM’ treatments provided mean (s.e.) percent reduction in trap catch of 99.5 (0.4) and 42.9 (10.1), respectively, for the redbanded leafroller,Argyrotaenia velutinana (Walker), and 90.4 (6.8) and 90.4 (1.3), respectively, for the obliquebanded leafroller,Choristoneura rosaceana (Harris).     

A dynamic model of motor basal ganglia functions

 Fast aiming movements were measured in a choice reaction paradigm in a healthy control group and in Parkinsonian patients. The patients were tested without (‘off ’) and with 3,4-dihydroxyphenylalanine (‘on’) (L-dopa) medication. The movement trajectories were used to estimate the parameters of a dynamic linear model. The model is based on the functional structure of the basal ganglia-thalamocortical circuit with direct and indirect pathways linking the putamen to the basal ganglia output nuclei (Albin et al. 1989). The output of the circuit is connected to a model for the motor neuron-musculo-skeletal system. The gain k _d for the direct pathway and the gain k _i for the indirect pathway were estimated. They were found to be significantly decreased for Parkinsonian patients in ‘off ’ compared with the control group. L-dopa therapy in Parkinsonian patients increased the gains of the direct and the indirect pathway almost to normal values which implies that the long-term dopamine level in the striatum was excitatory for the direct and for the indirect pathway. This result is restricted to movements of correct size. For movements of diminished size, which are typical for Parkinsonian patients, the model predicts that the dopamine level in the striatum is excitatory for the direct pathway but inhibitory for the indirect pathway. The simulated values for neuronal activities are in agreement with expected values according to the experimental data. The proposed model of the ‘motor’ basal ganglia thalamocortical circuit implies that information about biomechanical properties of the musculo-skeletal system is stored in the ‘motor’ basal ganglia-thalamocortical circuit, and that the basal ganglia are involved in computation of the desired movement amplitude. Received: 24 April 1996/Accepted in revised form: 25 February 1997     

Effects of different methods of acid hydrolysis on the nitrogen distribution in two soils

Summary An inorganic (Bainsville) and an organic (Farnham) soil were hydrolyzed by continuous and stepwise hydrolysis with hot 3N HCl for 1, 2, 3, 4, 11, 15, 18 and 24 h and continuously with hot 6N HCl for 24 h. The following nitrogen forms were determined: total N, hydrolyzable-N, amino acid-N, amino sugar-N and ammonia-N. Proportions of ‘unknown’ N were computed from these data. Continuous hydrolysis with 3N HCl yielded more amino acid-N and less ‘unknown’ N than did stepwise hydrolysis with the same acid strength. But continuous hydrolysis for 24 h with 6N HCl produced more amino acid-N and less ‘unknown’ N than did hydrolysis with 3N HCl by either method. It was estimated that 33 and 54% of the total N in the inorganic and organic soil, respectively, was protein-N. The ‘unknown’ N in the inorganic and organic soil constituted 51 and 37% of the total N, respectively. From our work it appears that the ‘unknown’ N is not proteinaceous. It can be readily degraded chemically and microbiologically to NH₃ and N-gases. More attention needs to be given to identifying the ‘unknown’ N which constitutes a large portion of the total soil-N. A more adequate knowledge of the chemical constitution of the ‘unknown’ soil N may lead to the development of technologies that will make more efficient use of the N in soils.     

Characterization of soluble thrombomodulin levels in patients with stage 3–5 chronic kidney disease

Abstract

Objective: This study aims to test the serum levels of soluble thrombomodulin (TM) in patients with chronic kidney disease (CKD)3–5 and to assess their connection with the different stages and severity of disease.

Methods: Sixty-seven patients with CKD are included, disease severity was evaluated accordingly to CKD staging and clinical data is collected. Nineteen healthy volunteers served as healthy controls. Serum soluble TM is analyzed by ELISA.

Results: The levels of soluble TM in all patients with CKD were significantly higher than those of healthy controls (p?<?0.001). CKD5 patients showed higher serum levels of soluble TM, in comparison to CKD4 patients (p?=?0.001), CKD3 patients (p?<?0.001), and healthy controls (p?<?0.001). The correlation analysis revealed significant correlation between serum soluble TM and disease severity (r?=?0.714, p?<?0.001). Serum soluble TM was found to be correlated with eGFR (r?=??0.766; p?<?0.001) and serum creatinine (r?=?0.778, p?<?0.001).

Conclusion: Soluble TM concentrations significantly increase in the CKD patients and are associated with the severity of the disease. Soluble TM may play critical roles in the development of CKD, as a biomarker of endothelial cells damage, anticoagulation and anti-inflammation.     

Impact of sulfur on density of <Emphasis Type="Italic">Tetranychus pacificus</Emphasis> (Acari: Tetranychidae) and <Emphasis Type="Italic">Galendromus occidentalis</Emphasis> (Acari: Phytoseiidae) in a central California vineyard

Sulfur is the oldest and most widely used fungicide in the vineyards of California, where it is used for control of powdery mildew (Uncinula necator [Schw.] Burr). For decades, sulfur use has been associated with outbreaks of Tetranychus pacificus McGregor (Acari: Tetranychidae) on cultivated grapes in the San Joaquin Valley. I undertook large-scale field studies to test this association, to evaluate the impact of sulfur on Galendromus occidentalis (Nesbit) (Acari: Phytoseiidae), a major predator of T. pacificus, and to determine if timing of sulfur applications with respect to grape bloom has an impact on T. pacificus density. The studies took place in a 32 ha vineyard in Fresno County, and all fungicide applications were made with commercial-scale equipment. In 1998 a ‘high sulfur’ treatment, a combination of wettable sulfur and sulfur dust, was compared to ‘low sulfur,’ in which demethylation inhibitor (DMI) fungicides partially substituted for sulfur. In 1999 treatments were ‘sulfur,’ ‘DMI,’ ‘sulfur pre-bloom’ (here sulfur was applied prior to grape bloom, in late May, and then DMIs were applied until mid-season) and ‘sulfur post-bloom’ (the reverse of ‘sulfur pre-bloom’). In each year, the T. pacificus population increase came after the end of fungicide applications, and results clearly show a relationship between sulfur use and T. pacificus density. In 1998, mean T. pacificus density was 2.7 times higher and mean G. occidentalis density 2.5 times higher in ‘high sulfur’ compared to ‘low sulfur.’ In 1999, the highest T. pacificus counts were in the ‘sulfur’ and ‘sulfur pre-bloom’ treatments, 4.8 times higher than ‘sulfur post-bloom’ and 2 times higher than ‘DMIs.’ Density of G. occidentalis was 2.3 times as high in ‘sulfur’ or ‘sulfur pre-bloom’ than ‘DMIs.’ The predator/prey ratio was not significantly different among treatments in 1998, but in 1999 it was highest in the ‘sulfur pre-bloom’ treatment. In 1999, density of Homeopronematus anconai (Baker) (Acari: Tydeidae) was 2.7 times higher in ‘sulfur pre-bloom’ compared to ‘sulfur,’ and higher by 2.7 times in ‘DMI’ compared to ‘sulfur post-bloom,’ suggesting a negative effect of sulfur on this tydeid. These results do not support the hypotheses that the cause of the increase in T. pacificus density is due to negative effects of sulfur on phytoseiids or tydeids. Rather, it appears that a plant-based explanation is likely, first, because of the differences in pre-bloom versus post-bloom sulfuring, and second, because of the long lag time between the end of the sulfur applications and the corresponding increase in spider mite density.     

Exogenous ethylene influences flower opening of cut roses (<Emphasis Type="Italic">Rosa hybrida</Emphasis>) by regulating the genes encoding ethylene biosynthesis enzymes

The purpose of this paper is to investigate the differential responses of flower opening to ethylene in two cut rose cultivars, ‘Samantha’, whose opening process is promoted, and ‘Kardinal’, whose opening process is inhibited by ethylene. Ethylene production and 1-aminocyclopropane-1-carboxylate (ACC) synthase and oxidase activities were determined first. After ethylene treatment, ethylene production, ACC synthase (ACS) and ACC oxidase (ACO) activities in petals increased and peaked at the earlier stage (stage 3) in ‘Samantha’, and they were much more dramatically enhanced and peaked at the later stage (stage 4) in ‘Kardinal’ than control during vasing. cDNA fragments of three Rh-ACSs and one Rh-ACO genes were cloned and designated as Rh-ACS1, Rh-ACS2, Rh-ACS3 and Rh-ACO1 respectively. Northern blotting analysis revealed that, among three genes of ACS, ethylene-induced expression patterns of Rh-ACS3 gene corresponded to ACS activity and ethylene production in both cultivars. A more dramatic accumulation of Rh-ACS3 mRNA was induced by ethylene in ‘Kardinal’ than that of ‘Samantha’. As an ethylene action inhibitor, STS at concentration of 0.2 mmol/L generally inhibited the expression of Rh-ACSs and Rh-ACO in both cultivars, although it induced the expression of Rh-ACS3 transiently in ‘Kardinal’. Our results suggests that ‘Kardinal’ is more sensitive to ethylene than ‘Samantha’; and the changes of Rh-ACS3 expression caused by ethylene might be related to the acceleration of flower opening in ‘Samantha’ and the inhibition in ‘Kardinal’. Additional results indicated that three Rh-ACSs genes were differentially associated with flower opening and senescence as well as wounding.     

Linkage mapping of two mutations that reduce phytic acid content of barley grain

 This study describes the inheritance and linkage map positions of two low phytic acid barley (Hordeum vulgare) mutations, lpa1-1 and lpa2-1, that dramatically reduce grain phytic acid content and increase inorganic seed phosphorus (P). Wide-cross, F₂ mapping populations were constructed by mating six-rowed varieties, ‘Steptoe’ and/or ‘Morex’, with two-rowed ‘Harrington’lpa donor lines homozygous for either lpa1-1 or lpa2-1. The barley lpa1-1 mutation showed normal inheritance patterns, whereas a deficiency of homozygous lpa2-1/lpa2-1 F₂ plants was observed. We identified a codominant, STS-PCR marker (aMSU21) that cosegregated with lpa1-1 in a population of 41 F₂ plants. The aMSU21 marker was then mapped to a locus on barley chromosome 2H, using a North American Barley Genome Mapping Project (NABGMP) doubled haploid population (‘Harrington’בMorex’). We determined that lpa2-1 is located within a recombination interval of approximately 30 cM between two AFLP markers that were subsequently mapped to barley chromosome 7H by integration with the same NABGMP population. Recent comparative mapping studies indicate conserved genetic map orders of several homologous molecular marker loci in maize and the Triticeae species that also show corresponding linkage to the biochemically similar lpa2 mutations of maize and barley. This observation suggests that barley and maize lpa2 mutations may affect orthologous genes. No such evidence for correspondence of the phenotypically similar lpa1 mutations of barley and maize has been revealed. Received: 22 September 1997 / Accepted: 2 December 1997     

Somatic embryogenesis from stigmas and styles of grapevine

Summary An in vitro protocol has been developed for callus indiction, somatic embryogenesis, and plant regeneration from stigma-style culture of grapevine. Four different grapevine cultivars (Vitis vinifera L.: cvs. ‘Bombino Nero’, ‘Greco di Tufo’, ‘Merlot’, and ‘Sangiovese’) were tested. Exlants were cultured on Nitsch and Nitsch medium (NN) supplemented with various combinations of 6-benzylaminopurine (BA: 4.5 and 9.0 μM) and β-naphthoxyacetic acid (NOA; 5.0 and 9.9 μM). Sucrose (88 mM) was used as the carbon source. Somatic embryogenesis was induced within 3–7 mo. after culture initiation. Even though explants of different origin (unfertilized ovules and anthers) regenerated somatic embryos, the higher embryogenic potential was observed in stigma and style explants, with the exception of ‘Merlot’, which regenerated somatic embryos only from unfertilized ovules. The percentages of stigma-style explants producing somatic embryos was 7% in ‘Bombino Nero’ (cultured on NN medium supplemented 9.0 μM BA and 9.9 μM NOA). 14% in ‘Greco di Tufo’ (4.5 μM BA and 9.9 μM NOA), and 8% in ‘Sangiovese’ (9.0 μM BA and 9.9 μM NOA). The presence of growth regulators (BA and NOA) in the medium was essential for induction of somatic embryogenesis. Plants were regenerated on hormone-free NN medium containing 88 mM sucrose.     

Hybrid origin of some ornamentals of Allium subgenus Melanocrommyum verified with GISH and RAPD

 Random amplified polymorphic DNA (RAPD) and genomic in situ hybridization (GISH) methods have been used to verify the hybridogenic origin and to identify the parental species of some ornamental cultivars in the subgenus Melanocrommyum of the genus Allium. The cultivars had been selected from seed obtained after uncontrolled pollination in breeders’ fields. The combination of GISH analysis with RAPD markers is very suitable for testing the hybridogenic origin of plants and to ascertain the parental species of the hybrids in such cases. As suspected, A. macleanii and A. cristophii are the parental species of ‘Globemaster’. The parental species of cultivar ‘Globus’ are A. karataviense and A. stipitatum, and not A. cristophii and A. giganteum as has been assumed on morphological grounds. Cultivars ‘Lucy Ball’ and ‘Gladiator’ are of hybrid origin, though only one of the parental species, A. hollandicum, could be confirmed. The cultivars ‘Purple Sensation’, ‘Mount Everest’, ‘White Giant’, ‘Michael H. Hoog’ and ‘Mars’ are not hybrids since neither GISH nor RAPD suggest the presence of a second genome. ‘Purple Sensation’ belongs to A. hollandicum, ‘Mount Everest’, ‘White Giant’ and ‘Mars’ to A. stipitatum,‘Michael H. Hoog’ to A. rosenorum. Received: 3 July 1997 / Accepted: 9 October 1997     

Analysis of hybrids of durum wheat with Thinopyrum junceiforme using RAPD markers

 The objective of this study was to detect the presence of alien chromatin in intergeneric hybrids of durum wheat (Triticum turgidum, 2n=4x=28; AABB genomes) with the perennial grass Thinopyrum junceiforme (2n=4x=28; J₁J₁J₂J₂) using RAPD markers. The first step was to identify amplification of species-specific DNA markers in the parental grass species and durum wheat cultivars. Initially, the genomic DNA of five grass species (Thinopyrum junceiforme, Th. bessarabicum, Lophopyrum elongatum, Leymus karataviensis and Elytrigia pycnantha) and selected durum cultivars (‘Langdon’, ‘Durox’, ‘Lloyd’, ‘Monroe’, and ‘Medora’) was screened with 40 oligonucleotide primers (nano-mers). Three oligonucleotides that amplified DNA fragments specific to a grass species or to a durum cultivar were identified. Primer PR21 amplified DNA fragments specific to each of the five durum cultivars, and primers PR22 and PR23 amplified fragments specific to each of the grass species. Intergeneric hybrids between the durum cultivars ‘Langdon’, ‘Lloyd’ and ‘Durox’ and Th. junceiforme, and their backcross (BC) progeny were screened with all 40 primers. Six primers amplified parent-specific DNA fragments in the F₁ hybrids and their BC₁ progeny. Three primers, PR22, PR23 and PR41, that amplified Th. junceiforme DNA fragments in both F₁ and BC₁ were further analyzed. The presence of an amplified 1.7-kb Th. junceiforme DNA fragment in the F₁ hybrids and BC₁ progeny was confirmed using Southern analysis by hybridization with both Th. junceiforme genomic DNA and Th. junceiforme DNA amplified with primer PR41. With the exception of line BC₁F₂ no. 5, five selfed progeny of BC₁ and a BC₂ of line 3 (BC₁F₂ no. 3בLloyd’) from a cross of ‘Lloyd’×Th. junceiforme showed the presence of the 1.7-kb DNA fragment. All selfed BC₁ and BC₂ lines retained the 600-bp fragment that was confirmed after hybridization with Th. junceiforme DNA amplified with primer PR22. Other experiments using RFLP markers also showed the presence of up to seven Th. junceiforme DNA fragments in the F₁ hybrids and their BC progeny after hybridization with Th. junceiforme DNA amplified with primer PR41. These studies show the usefulness of molecular markers in detecting alien chromatin/DNA fragments in intergeneric hybrids with durum wheat. Received: 21 November 1996 / Accepted: 21 March 1997     

Genetic transformation of three sweet orange cultivars from explants of adult plants

The difficulty in adult tissue genetic transformation in woody species is still an obstacle to be overcome, including in most sweet orange cultivars of the Brazilian citrus industry. This work reports that, after in vitro culture adjustments, transgenic adventitious buds of ‘Hamlin’, ‘Pêra’, and ‘Valencia’ sweet oranges (Citrus sinensis L. Osbeck) were recovered using adult material as explant source, in genetic transformation experiments via Agrobacterium tumefaciens. The transgenic buds were identified by the GUS histochemical analysis and confirmed by PCR analysis, which indicated the presence of an amplified fragment of 817 bp corresponding to the uidA gene sequence. The efficiencies of genetic transformation for ‘Hamlin’, ‘Pêra’, and ‘Valencia’ sweet orange cultivars were 2.5, 1.4, and 3.7%, respectively. Media supplemented with auxins and cytokinins during co-culture, and media with high concentrations of cytokinins (3 mg L⁻¹) during transgenic selection led to the transformation and, consequently, the regeneration of adequate number of adventitious buds for the three cultivars. The use of sonication during the explant disinfection was not effective to reduce endophytic contamination and reduced transformation efficiency.     

Effect of genotype and medium composition on linseed (<Emphasis Type="Italic">Linum usitatissimum</Emphasis>) ovary culture

Breeding linseed (Linum usitatissimum L.) using haploid techniques allows breeders to develop new cultivars in a shorter time period. Many research groups successfully created new linseed genotypes through anther culture; however ovary culture has been the subject of only a few earlier studies. In the present study, the effect of genotype and growth regulators combination on callus induction and shoots regeneration in ovary culture of nine commercially important linseed cultivars was investigated. Ovaries were cultured on modified MS medium supplemented with three different combinations of plant growth regulators. Variable callogenic responses were expressed by all of the genotypes tested on different induction media. The results suggested that specific combination of growth regulators for callus induction must be designed for each genotype. Shoot regeneration from ovary derived callus is a critical phase of the whole gynogenetic process. Differences in adventitious shoot formation frequency among genotypes were demonstrated and four responsive genotypes have been selected. Ovary derived callus from cultivar ‘Mikael’ manifested the highest adventitious shoot formation frequency with a high number of shoots per explant. The optimum ratio of growth regulators for shoot regeneration was shown to depend on the genotype. Cultivars ‘Linola’, ‘Mikael’ and ‘Szaphir’ showed the highest shoot regeneration frequency when callus had originated on induction medium supplemented with 2 mg L⁻¹ BAP and 2 mg L⁻¹ NAA, while combination of 1 mg L⁻¹ BAP and 2 mg L⁻¹ IAA promoted shoot formation in ovary-derived callus of ‘Barbara’. The highest rate of shoots per explant has been obtained in second subculture.     

Mapping and proteomic analysis of albumin and globulin proteins in hexaploid wheat kernels (Triticum aestivum L.)

Albumins and globulins of wheat endosperm represent 20% of total kernel protein. They are soluble proteins, mainly enzymes and proteins involved in cell functions. Two-dimensional gel immobiline electrophoresis (2DE) (pH 4-7) × SDS-Page revealed around 2,250 spots. Ninety percent of the spots were common between the very distantly related cultivars ‘Opata 85’ and ‘Synthetic W7984’, the two parents of the International Triticeae Mapping Initiative (ITMI) progeny. ‘Opata’ had 130 specific spots while ‘Synthetic’ had 96. 2DE and image analysis of the soluble proteins present in 112 recombinant inbred lines of the F9-mapped ITMI progeny enabled 120 unbiased segregating spots to be mapped on 21 wheat (Triticum aestivum L. em. Thell) chromosomes. After trypsic digestion, mapped spots were subjected to MALDI-Tof or tandem mass spectrometry for protein identification by database mining. Among the ‘Opata’ and ‘Synthetic’ spots identified, many enzymes have already been mapped in the barley and rice genomes. Multigene families of Heat Shock Proteins, beta-amylases, UDP-glucose pyrophosphorylases, peroxydases and thioredoxins were successfully identified. Although other proteins remain to be identified, some differences were found in the number of segregating proteins involved in response to stress: 11 proteins found in the modern selected cultivar ‘Opata 85’ as compared to 4 in the new hexaploid `Synthetic W7984’. In addition, ‘Opata’ and ‘Synthetic’ differed in the number of proteins involved in protein folding (2 and 10, respectively). The usefulness of the mapped enzymes for future research on seed composition and characteristics is discussed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Resistance to Phytophthora fragariae var. fragariae in strawberry: the Rpf2 gene

  Phytophthora fragariae var. fragariae is the causal agent of red stele (red core) root rot in strawberry (Fragaria spp.). The inheritance of resistance to one isolate of this fungus was studied in 12 segregating populations of F.×ananassa derived from crosses between four resistant cultivars (‘Climax’, ‘Redgauntlet’, ‘Siletz’, and ‘Sparkle’) and three susceptible cultivars (‘Blakemore’, ‘Glasa’, and ‘Senga’ Sengana’). The analysis clearly supports the hypothesis of a single segregating dominant resistance gene. It is proposed that this gene be designated Rpf2. Received 12 November 1996 / Accepted: 22 November 1996     

Localization of the rice stripe disease resistance gene, Stv-bi, by graphical genotyping and linkage analyses with molecular markers

 We used graphical genotyping and linkage analyses with molecular markers to determine the chromosomal location of the rice stripe disease resistance gene, Stv-b ⁱ . The stripe resistance gene from the indica rice (Oryza sativa) cv ‘Modan’ was introgressed into several Japanese rice varieties. We found 4 RFLP markers in ‘Modan’, five susceptible parental rice varieties (‘Norin No. 8’, ‘Sachihikari’, ‘Kanto No. 98’, ‘Hokuriku No.103’ and ‘Koganebare’) and four resistant progeny varieties (‘St. No. 1’, ‘Aichi No. 6’, ‘Aoisora’ and ‘Asanohikari’). Graphical genotyping of the resistant progeny revealed a chromosomal segment ascribable to ‘Modan’ and associated with stripe resistance. The chromosomal segment from ‘Modan’ was located at 35.85 cM on chromosome 11. Linkage analysis using 120 F₂ individuals from a cross between ‘Koshihikari’ (susceptible) and ‘Asanohikari’ (resistant) revealed another 8 RFLP markers in the same chromosome. We performed a bioassay for rice stripe resistance in F₃ lines of the F₂ individuals using infective small brown planthoppers and identified an 1.8-cM segment harboring the rice stripe disease resistance gene, Stv-b ⁱ , between XNpb220 and XNpb257/ XNpb254. Furthermore, Stv-b ⁱ was linked by 0.0 cM to a RFLP marker, ST10, which was developed on the basis of the results of RAPD analysis. These DNA markers near the Stv-b ⁱ locus may be useful in marker-assisted selection and map-based cloning of the Stv-b ⁱ gene. Received: 26 September 1997 / Accepted: 4 November 1997