Monitoring the viability of citrus rootstocks seeds stored under refrigeration

The citrus nursery tree is produced through the bud grafting process, in which rootstock is usually grown from seed germination. The objective of this research was to evaluate, in two dissimilar environmental conditions, the viability and polyembryony expression of five citrus rootstocks seeds stored in different periods under refrigeration. The rootstock varieties evaluated were: Rangpur lime (Citrus limonia Osb. cv. Limeira), Trifoliate orange (Poncirus trifoliata Raf. cv. Limeira), Citrumelo (P. trifoliata x C. paradisi Macf. cv. Swingle), Sunki mandarin (C. sunki Hort. ex Tanaka) and Volkamer lemon (C. volkameriana Ten. & Pasq. cv. Catania 2). The experimental design was the randomized blocks in a 11 x 5 x 2 factorial scheme, evaluating from time zero to the tenth month of storage, the five varieties of rootstock in two environments: germination and growth B.O.D type chamber (Biological Oxygen Demand - Eletrolab Brand Model FC 122) at 25 °C; and greenhouse seedbed with partial temperature control (22 °C to 36 °C) and humidity control (75-85%). The plot had 24 seeds in four replicates, using trays with substrate in greenhouse and Petri dishes with filter paper in B.O.D. chamber. The seed germination rate and polyembryony expression were evaluated monthly. It was concluded that Trifoliate and Citrumelo Swingle seeds can be stored for up to seven months, while Volkamer lemon, Rangpur lime and Sunki seeds can be stored for up to ten months. The polyembryony expression rate was slightly higher when measured in greenhouse than in B.O.D. chamber and remained stable in both environments until the seventh month, from which dropped sharply. Citrumelo Swingle seeds expressed the highest polyembryony rate (18.8%), followed by Rangpur lime and Volkamer lemon (average value of 13.7%), Sunki (9.4%) and Trifoliate (3.2%). Despite some differences among varieties, the viability of rootstock stored seeds can be monitored either in the greenhouse or in B.O.D. germination chamber, the latter being the faster and more suitable method.

Conservation of Campomanesia adamantium (CAMB.) O. berg seeds in different packaging and at varied temperatures

This article aims at evaluating the effects of different packaging and varied storage temperatures on the germination potential of seeds of Campomanesia adamantium Camb. O. Berg. The seeds were packaged in glass, aluminum foil and plastic containers, or maintained inside intact fruits at 5, 10 and 15 ºC during 0, 7, 14 and 21 days. After these periods the seeds were sown in Germitest® germination paper and maintained in incubation chambers at 25 ºC under constant white light for 42 days. Seed moisture contents were evaluated both before and after storage, as well as germination percentages, germination speed index, root and aerial portion of seedlings lengths, and total dry weights. All possible combinations of packing materials, temperatures and storage times were tested, with four repetitions of 25 seeds for each treatment. C. adamantium seeds showed initial water contents of 31.5%. Glass and aluminum packaging were efficient at maintaining the water content of the seeds, and provided greater germination speed index than the other packaging materials. Germination percentages, seedlings lengths and dry weights did not vary among the different temperatures tested. C. adamantium seeds can be stored for up to 21 days at temperatures between 5 and 15 ºC without altering their physiological quality. In terms of cost-benefit efficiencies, these seeds can be stored without significant damage for 21 days while still inside the fruits at temperatures of 5, 10 or 15 ºC.

Effect of GA3, KNO3, and removing of basal point of seeds on germination of sweet granadilla (Passiflora ligularis Juss) and yellow passion fruit (Passiflora edulis f. flavicarpa)

Passiflora seeds germinate erratically presenting difficulties for their handling in a greenhouse. The effect of removing of basal point of seeds (RB) and pre-imbibition of seeds of sweet granadilla and yellow passion fruit in 50, 100, 200, and 400 mg mL-1 solutions of gibberellic acid (GA3) or 0.1% KNO3 solution was studied. The experiment was conducted in greenhouses in La Plata, Colombia. Two accessions PrJ1 and PrJ2 of sweet granadilla were evaluated. There were calculated the final percentage of germination (PG), mean germination time (MGT), and the mean germination rate (MGR). The leaf area and dry mass of seedlings were measured 22 days after sowing (das); with this data, specific leaf area and relation root/shoot were calculated. In all cases, the highest germination percentages were achieved treating seeds with KNO3 (89, 92, and 87% for yellow passion fruit, PrJ2, and PrJ1, respectively), but the increase in MGR (3.3 germinated seeds per day) and the decrease in MGT (16 days) were only significant for PrJ1. RB had a significant reduction of PG in all cases (28, 12, and 33% for passion fruit, PrJ2 and PrJ1, respectively). With the increase in the concentration of GA3, PG was reduced for two accessions of sweet granadilla, for yellow passion fruit this trend was not clear, no treatment with GA3 showed significant differences with the control. Leaf area (24.07 cm2) and dry mass of seedlings (135 mg) were significantly higher than seeds previously treated with KNO3 only for PrJ1.The solution of KNO3 0,1% is recommended to improve the germination and initial growth of granadilla seedlings.

Biotechnology applied to Annona species: a review

Annonaceae is an ancient family of plants including approximately 50 genera growing worldwide in a quite restricted area with specific agroclimatic requirements. Only few species of this family has been cultivated and exploited commercially and most of them belonging to the genus Annona such as A. muricata, A. squamosa, the hybrid A. cherimola x A. squamosa and specially Annona cherimola: the cherimoya, commercially cultivated in Spain, Chile, California, Florida, México, Australia, Ecuador, Peru, Brazil, New Zealand and several countries in South and Central America. The cherimoya shows a high degree of heterozygosis, and to obtain homogeneous and productive orchards it is necessary to avoid the propagation by seeds of this species. Additionally, the traditional methods of vegetative propagation were inefficient and inadequate, due to the low morphogenetic potential of this species, and the low rooting rate. The in vitro tissue culture methods of micropropagation can be applied successfully to cherimoya and other Annona sp to overcome these problems. Most of the protocols of micropropagation and regeneration were developed using the cultivar Fino de Jete, which is the major cultivar in Spain. First it is developed the method to micropropagate the juvenile material of cherimoya (ENCINA et al., 1994), and later it was optimized a protocol to micropropagate adult cherimoya genotypes selected by outstanding agronomical traits (PADILLA and ENCINA, 2004) and further it was improved the process through micrografting (PADILLA and ENCINA, 2011).At the present time we are involved in inducing and obtaining new elite genotypes, as part of a breeding program for the cherimoya and other Annonas, using and optimizing different methodologies in vitro: a) Adventitious organogenesis and regeneration from cellular cultures (ENCINA, 2004), b) Ploidy manipulation of the cherimoya, to obtain haploid, tetraploid and triploid plants (seedless), c) Genetic transformation, for the genes introduction to control the postharvest processes and the genes introduction to provide resistance to pathogen and insects and d) Micropropagation and regeneration of other wild Annona or related Annonaceae species such as: Annona senegalensis, A. scleroderma, A. montana, A. reticulata, A. glabra, A. diversifolia and Rollinia sp.

Soluble sugars and germination of Annona emarginata (Schltdl.) H. Rainer seeds submitted to immersion in GA3 up to different water contents

The objective of this study was to evaluate the effect of different water contents achieved by Annona emarginata (Schltdl.) H. Rainer seeds during immersion in GA3 solutions, in variation of soluble sugars levels and germination. Seeds with 10% of initial water content were submitted to imbibition in GA3 solutions with concentrations of 0; 250; 500; 750 and 1000 mg L-1 and when they reached the water content of 15%, 20%, 25%, 30% and 35%, the quantification of soluble sugars levels and germination test were performed. Seeds immersed up to they reach 15% of water with GA3 and immersed up to the water acquisition of 20% without GA3, presented higher soluble sugars levels and germination percentage, which were decreased when the seeds reached 30% and 35% of water, independently of the presence of the plant growth regulator. It was conclude that different water contents reached by the seeds in immersion treatments with GA3 affect the soluble sugars levels and germination percentage of Annona emarginata seeds. Thus, in treatments with Annona emarginata, the seeds must remain immersed in water without GA3 up to they reach 20% of water, as higher water contents (35%) reduce the soluble sugars levels and the seed germination percentage.

Water uptake by Annona diversifolia Saff. and A. purpurea Moc. & sessé ex dunal seeds (Annonaceae)

Annonaceae seeds are known by presenting dormancy mechanisms, whose reports ranging from coating impermeable to the physiological dormancy. By this way, the present study aimed to evaluate water uptake in Annona diversifolia Saff and Annona purpurea Moc & Sessé ex Dunal seeds. For this study, seeds were placed under immersion in distilled water, and used four replicates of 25 seeds of each species, which were weighed during the 480 hours that were immersed. To determine the place of purchase of water, Annona diversifolia seeds were sealed with paraffin at different locations. Based on the results, seeds from both species reached the phases I and II of water uptake, which indicates they are not hard; however, germination (Phase III) was not reached. Annona diversifolia seeds completed Phase I with, 50h and Annona purpurea with 70h from imbibitions begin, which shows that even slowly, water is acquire.

Mono and diterpenes from seeds of Xylopia sericea

A monoterpene, 3beta,6beta-dihydroxy-p-menth-1-ene has been isolated from the seeds of Xylopia sericea along with four kaurane, one beyerene, one atisene and four trachylobane diterpenoids, including the trachyloban-18- and 19-methyl esters. The X-ray crystal structure of methyl ent-trachyloban-18-oate was determined in order to make an unambiguous distinction between the 18- and 19-esters. The 13C NMR data for ent-15alpha-hydroxy-trachyloban-19-oic acid has been revised.

Chemical composition of umbu (Spondias tuberosa Arr. Cam) seeds

The umbu tree (Spondias tuberosa Arr. Cam) is an important fruit tree the economy of the semi-arid northeastern region of Brazil. With the objective of finding use for the seeds, physical and chemical characterizations of the seeds from 2 cultivars in 2 maturation stages were carried out and their fatty acid and mineral profiles determined. The results showed no differences between the seeds analyzed. The yield was about 10% and the dimensions as follows: length from 1.48 to 2.11 cm and width from 0.76 to 1.16 cm. The average lipid content was 55% of which 69% was unsaturated and the average protein content was 24%. The seeds were a good source of the following minerals: P, K, Mg, Fe and Cu. The overall results indicated that the oil or the seeds could be used for food stuffs if no toxic agents were found.

Highly improved chromium (III) uptake capacity in modified sugarcane bagasse using different chemical treatments

The present paper focuses on improving chromium (III) uptake capacity of sugarcane bagasse through its chemical modification with citric acid and/or sodium hydroxide. The chemical modifications were confirmed by infrared spectroscopy, with an evident peak observed at 1730 cm-1, attributed to carbonyl groups. Equilibrium was reached after 24 h, and the kinetics followed the pseudo-second-order model. The highest chromium (III) maximum adsorption capacity (MAC) value was found when using sugarcane bagasse modified with sodium hydroxide and citric acid (58.00 mg g-1) giving a MAC value about three times greater (20.34 mg g-1) than for raw sugarcane bagasse.

Chemical characterization by GC-MS and phytotoxic potential of non-polar and polar fractions of seeds of Dioteryx odorata (Aubl.) Willd. from Venezuelan regions

Dipteryx odorata (Aubl.) Willd. is a tall arboreal species native to Central and Northern South America. This paper describes the chemical characterization and phytotoxic potential of polar and non-polar extracts from D. odorata seeds. Structural determinations were accomplished by chemical derivatization and analyzed by GC/MS. The chemical composition of the non-polar fraction (hexane and dichloromethane) presented fatty acids as major constituent. Medium polar and polar fractions (ethyl acetate and ethanol: water) contained carboxylic acid and high 6,7-Dyhidroxycoumarin-β-D-glucopyranoside content, not previously reported for seeds of D. odorata. Extracts showed a significant level of phytotoxic activity, correlated to the content of coumarin derivatives, predominantly in the polar fraction.

Improved method to obtain pfaffic acid as a marker for quality control

Pfaffic acid, a marker of Hebanthe eriantha (Brazilian ginseng), was first isolated in 1983 but is not yet commercially available. This lack of availability compromises the quality control of this plant and its derivatives. This paper proposes a process for pfaffic acid isolation from roots of H. eriantha at a purity suitable for analytical purposes. The steps involved in this process included extraction, hydrolysis, fractionation and purification by preparative HPLC. This process led to isolation of pfaffic acid with a chromatographic purity of 98.5% in a 0.25% yield from dried roots of H. eriantha; this yield is more than forty times higher than that of the current method in the literature.

Evaluation of plant growth-promoting and bioprotecting rhizobacteria on wheat crop

Experiments were carried out under laboratory, growth chamber, and field conditions to evaluate the effect of Plant growth-promoting and bioprotecting rhizobacteria (PGPBR) seed treatment on seed pathogens, seed germination, plant growth, and grain yield of wheat (Triticum aestivum). Most of the PGPBR strongly reduced the recovery of the pathogens from infected wheat seeds. All treatments, except the chemical iprodione + thiram, significantly promoted plant growth over the nontreated control. Psudomonas putida biotype A (11) and P. agglomerans (14) showed the greatest effects. Field experiments, carried out at two locations, indicated that all treatments, except P. chlororaphis (42), significantly increased seedling emergence of wheat . In Pato Branco, PR, P. putida biotype A (11) and P. putida biotype B (44) presented the best results, both being superior to fungal biological and chemical treatments. In Passo Fundo P. putida biotype A (11) and P. putida biotype B (17 and 44) significantly improved yield over the nontreated control. Yield increases of these three PGPBR were similar to the chemical treatment iprodione + thiram. In Pato Branco, P. putida biotype A (11) and P. putida biotype B (17), as well as the chemical treatment, provided significant increase over the nontreated control. Yield increases by the PGPBR varied from 18% to 22% in Passo Fundo and from 27% to 28% in Pato Branco.

Detection of drechslera avenae in oat seeds

The fungus Drechslera avenae, the causal agent of Helminthosporium leaf spot on oats (Avena sativa), survives as mycelium in crop residues and in infected seeds. In trials carried out in the laboratory, ten methods were evaluated for their efficiency to detect D. avenae in oat seeds. In each experiment, groups of two or three methods were compared to a standard protocol, in which seeds were placed in Petri dishes containing the Reis selective medium and incubated at 25±2 °C for ten days. Data were submitted to analysis of variation and the means of the methods were compared using the Dunnett test at the 5% significance level. Overall, the highest levels of seed infection by D. avenae were observed on oat seeds plated in the osmotic, the oat-agar and the Reis media, or on seeds subjected to heat treatment previous to incubation in malt-agar. Therefore, these methods should be recommended for detection of D. avenae in oat seed testing.

Use of semi-seletive media for detection of Sclerotinia sclerotiorum on bean and soybean seeds

This work was aimed at evaluating the possibility of using bromophenol blue as an indicator for detecting the presence of Sclerotinia sclerotiorum in the seeds of dry-beans (Phaseolus vulgaris) and soybean (Glycine max), through incubation of the seeds on an agar medium and "blotter" substrates. The seeds were artificially inoculated with four S. sclerotiorum isolates, plated on the agar medium, named Neon, and on modified Neon agar media all incubated at 14 and 20 ºC for seven days in the dark. Half of the seeds inoculated were surface desinfested prior to plating on the medium. The seeds showing change of colour in the medium, from blue to light yellow, as well as formation of typical mycelium and sclerotia in some cases, were considered to be infected or contaminated by S. sclerotiorum. The two incubation temperatures compared did not show significant (P<0.05) differences in detection level for most of the isolates tested on the different media. According to results obtained in this study, the Neon agar medium with incubation at 14 or 20 ºC has proved to be a reliable and quick method for the detection of S. sclerotiorum mycelium in naturally infected seeds of bean and soybean.

Pathotype diversity of Pyricularia grisea from improved upland rice cultivars in experimental plots

A study was undertaken to examine the pathogenic diversity of Pyricularia grisea isolates retrieved from 14 upland rice (Oryza sativa) cultivars in experimental plots during a period of five years. Inoculations were performed on 32 genotypes with 85 monoconidial isolates under controlled greenhouse conditions. Based on the reaction pattern of eight international differentials, eleven pathotypes of P. grisea were identified. The predominant international races or pathotypes were IB-9 (56.4%), IB-1 (16.4%) and IB-41 (11.8%). A set of eight commercial upland rice cultivars ('Carajás', 'Confiança', 'Maravilha', 'Primavera', 'Progresso', 'Caiapó', 'IAC-47', 'IAC-201') was utilized as additional differentials for describing the virulence pattern of P. grisea. Twenty-six Brazilian pathotypes were identified on the basis of disease reaction on these differentials, in contrast to the 11 international pathotypes. The most predominant Brazilian pathotypes, BB-21 and BB-41 were represented by 28.2% and 17.6% of the isolates tested, respectively. Isolates virulent and avirulent to cultivar 'Primavera' were encountered within the pathotype IB-1. Utilizing Brazilian cultivars as differentials, the 14 isolates of the pathotype IB-1could be further classified into eight local pathotypes, BB-41, BB-13, BB-21, BB-9, BB-29, BB-61, BD-9 and BG-1. Virulence to improved rice cultivars 'Canastra', 'Confiança', 'Carisma', 'Maravilha', 'Primavera' and 'Bonança' was frequent in pathogen population. Some of the Brazilian pathotypes that showed differential reaction on commercial rice cultivars could be utilized for incorporating resistance genes in susceptible cultivars improved for grain quality, by conventional breeding methods.