Seed-specific silencing of OsMRP5 reduces seed phytic acid and weight in rice.

Phytic acid (PA) is poorly digested by humans and monogastric animals and negatively affects human/animal nutrition and the environment. Rice mutants with reduced PA content have been developed but are often associated with reduced seed weight and viability, lacking breeding value. In the present study, a new approach was explored to reduce seed PA while attaining competitive yield. The OsMRP5 gene, of which mutations are known to reduce seed PA as well as seed yield and viability, was down-regulated specifically in rice seeds by using an artificial microRNA driven by the rice seed specific promoter Ole18. Seed PA contents were reduced by 35.8-71.9% in brown rice grains of transgenic plants compared to their respective null plants (non-transgenic plants derived from the same event). No consistent significant differences of plant height or number of tillers per plant were observed, but significantly lower seed weights (up to 17.8% reduction) were detected in all transgenic lines compared to null plants, accompanied by reductions of seed germination and seedling emergence. It was observed that the silencing of the OsMRP5 gene increased the inorganic P (Pi) levels (up to 7.5 times) in amounts more than the reduction of PA-P in brown rice. This indicates a reduction in P content in other cellular compounds, such as lipids and nucleic acids, which may affect overall seed development. Put together, the present study demonstrated that seed specific silencing of OsMRP5 could significantly reduce the PA content and increase Pi levels in seeds; however, it also significantly lowers seed weight in rice. Discussions were made regarding future directions towards producing agronomically competitive and nutritionally valuable low PA rice.

Seed production characters and germination performance of reed canary grass in Finland

Selostus: Ruokohelven siementuotanto-ominaisuudet ja itävyys Suomessa

Spore germination and development of young mycelia in some Rhizopogon species

Spore germination in Rhizopogon abietis, R. luteolus, R. roseolus and R. villosulus was induced in the presence of Rhodotorula glutinis and activated charcoal, in agar medium (N6:5). In one R. roseolus sample, 51% of spores germinated within 35 days, allowing observation of the course of spore germination and the different developmental patterns of homokaryotic mycelia. In these plates, spores showed two times of germination. The spores that germinated early produced an apical germ tube. Later other spores germinated in proximity to young mycelium, by forming a germ vesicle. One of the hyphal growth patterns obtained (interruption-swelling-ramification) is similar to that reported for other fungi. With this technique for inducing spore germination, it is possible to obtain enough monosporic cultures to perform mating tests. Key words: Basidiomycotina, Hypogeous, Monosporic Cultures.

Biochemical changes associated to soybean seeds osmoconditioning during storage

A work was carried out with the purpose of verifying the biochemical changes associated to soybean (Glycine max (L.) Merrill) seeds osmoconditioning. Seeds of the UFV 10, IAC 8 and Doko RC cultivars harvested at R8 development stage and submitted to different treatments were used. The biochemical evaluations were performed during seed storage, after the hydration-dehydration process. Initially, seeds were osmoconditioned in a polyethylene glycol (PEG 6000) solution, with the osmotic potential of -0.8 MPa and 20ºC, for a period of four days. After that, seeds were dried back until the initial moisture content (10-11%) and stored in natural conditions for three and six months. Two controls were used: untreated seeds (dry seeds) and water soaked seeds. Seed changes in protein and lipid, hexanal accumulation and fatty acids contents were evaluated. The results showed that seed storage under laboratory natural conditions caused reduction in protein, lipid and polyunsaturated fatty acids content and promoted hexanal production. Storage periods reduced protein levels for all treatments, however the PEG 6000 treatment showed lower protein reduction. The soybean seed storage increased hexanal production, but hexanal levels were smaller with osmoconditioning comparing to the other imbibition treatments.

Composition, ileal amino acid digestibility and nutritive value of organically grown legume seeds and conventional rapeseed cakes for pigs

Selostus: Palkoviljojen ja rypsipuristeiden koostumus, aminohappojen ohutsuolisulavuus sekä rehuarvo sikojen ruokinnassa

Bioassay for detection of transgenic soybean seeds tolerant to glyphosate

Glyphosate is a systemic, nonselective, postemergence herbicide that inhibits growth of both weeds and crop plants. Once inside the plant, glyphosate interferes with biosynthesis of aromatic amino acids phenylalanine, tyrosine, and tryptophan, by inhibiting the activity of 5enolpyruvylshikimate-3-phosphate synthase (EPSPS), a key enzyme of the shikimate pathway. The objective of this work was to develop a simple, effective and inexpensible method for identification of transgenic soybean tolerant to glyphosate. This technique consisted in germinating soybean seeds in filter paper moistened with 100 to 200 muM of glyphosate. Transgenic soybean seeds tolerant to glyphosate germinated normally in this solution and, between 7 and 10 days, started to develop a primary root system. However non-transgenic seeds stopped primary root growth and emission of secondary roots.

Nonenzymatic lipid peroxidation reprograms gene expression and activates defense markers in Arabidopsis tocopherol-deficient mutants.

Tocopherols (vitamin E) are lipophilic antioxidants that are synthesized by all plants and are particularly abundant in seeds. Two tocopherol-deficient mutant loci in Arabidopsis thaliana were used to examine the functions of tocopherols in seedlings: vitamin e1 (vte1), which accumulates the pathway intermediate 2,3-dimethyl-5-phytyl-1,4-benzoquinone (DMPBQ); and vte2, which lacks all tocopherols and pathway intermediates. Only vte2 displayed severe seedling growth defects, which corresponded with massively increased levels of the major classes of nonenzymatic lipid peroxidation products: hydroxy fatty acids, malondialdehyde, and phytoprostanes. In the absence of pathogens, the phytoalexin camalexin accumulated in vte2 seedlings to levels 100-fold higher than in wild-type or vte1 seedlings. Similarly, gene expression profiling in wild-type, vte1, and vte2 seedlings indicated that increased levels of nonenzymatic lipid peroxidation in vte2 corresponded to increased expression of many defense-related genes, which were not induced in vte1. Both biochemical and transcriptional analyses of vte2 seedlings indicate that nonenzymatic lipid peroxidation plays a significant role in modulating plant defense responses. Together, these results establish that tocopherols in wild-type plants or DMPBQ in vte1 plants limit nonenzymatic lipid peroxidation during germination and early seedling development, thereby preventing the inappropriate activation of transcriptional and biochemical defense responses.

The IBO germination quantitative trait locus encodes a phosphatase 2C-related variant with a nonsynonymous amino acid change that interferes with abscisic acid signaling.

Natural genetic variation is crucial for adaptability of plants to different environments. Seed dormancy prevents precocious germination in unsuitable conditions and is an adaptation to a major macro-environmental parameter, the seasonal variation in temperature and day length. Here we report the isolation of IBO, a quantitative trait locus (QTL) that governs c. 30% of germination rate variance in an Arabidopsis recombinant inbred line (RIL) population derived from the parental accessions Eilenburg-0 (Eil-0) and Loch Ness-0 (Lc-0). IBO encodes an uncharacterized phosphatase 2C-related protein, but neither the Eil-0 nor the Lc-0 variant, which differ in a single amino acid, have any appreciable phosphatase activity in in vitro assays. However, we found that the amino acid change in the Lc-0 variant of the IBO protein confers reduced germination rate. Moreover, unlike the Eil-0 variant of the protein, the Lc-0 variant can interfere with the activity of the phosphatase 2C ABSCISIC ACID INSENSITIVE 1 in vitro. This suggests that the Lc-0 variant possibly interferes with abscisic acid signaling, a notion that is supported by physiological assays. Thus, we isolated an example of a QTL allele with a nonsynonymous amino acid change that might mediate local adaptation of seed germination timing.

Environmental and genetic variation of isoflavone content of soybean seeds grown in Brazil

The objective of this work was to evaluate isoflavone concentrations in seeds of different Brazilian soybean cultivars grown in a range of locations and environmental conditions in Brazil. Seeds of 233 cultivars grown in Ponta Grossa, PR, Brazil, during the 2001/2002 soybean season, and of 22 cultivars sown in different locations of Brazilian Northeast, Southeast on South regions were analyzed for total isoflavones, including daidzin, glycitin, genistin and acetylgenistin. The total isoflavones ranged from 12 mg 100 g-1 (cv. Embrapa 48) to 461 mg 100 g-1 (cv. CS 305) among the 233 cultivars grown in Ponta Grossa, and the differences among them are due to genetic effects since all cultivars were grown and collected at the same locatation and year. This is an indication of the possibility of breeding for isoflavone content. Differences in isoflavone content observed in the cultivars grown in different locations permit the selection of locations for optimum isoflavone content (low or high), depending on the uses of soybean. In the Northeast region (5-8°S), higher concentrations of total isoflavones were observed at São Raimundo das Mangabeiras (232 mg 100 g-1) and Tasso Fragoso (284 mg 100 g-1) municipalities, and in the South (23-30°S), isoflavones were higher in Guarapuava, Canoinhas, Vacaria and Campos Novos municipalities, ranging from 130 to 409 mg 100 g-1.

Desiccation, postharvest maturity and seed aging of tall oat-grass

The objectives of this work were to determine whether and at which seed physiological maturity stage the diquat desiccant affects the tall oat-grass (Arrhenatherum elatius) seed quality and yield, as well as the proper storage period between harvest and sowing. Diquat desiccation was evaluated in applications during milk or dough seed maturation stages. Seeds conventionally produced and stored under traditional storage conditions were used for the analyses. Seed samples were drawn every 30th day after harvest (DAH). After the 240th DAH, samples were drawn every 90th day up the to 690th DAH. The highest yield were obtained by desiccation applied at the beginning of the seed dough stage, with a satisfactory seed quality. Both final germination and seedling growth parameters achieved their maximum values between 180th and 240th DAH. Oat-grass seeds preserved satisfactory level of final germination (75%) up to 420th DAH. The application of diquat desiccant at the beginning of seed dough maturity stage can be a good solution for seed production of tall oat-grass. Early spring is the best sowing period for freshly harvested seeds of tall oat-grass regarding germination and seedling growth.

Potential of antimicrobial volatile organic compounds to control Sclerotinia sclerotiorum in bean seeds

The objective of this work was to evaluate the potential of an artificial mixture of volatile organic compounds (VOCs), produced by Saccharomyces cerevisiae, to control Sclerotinia sclerotiorum in vitro and in bean seeds. The phytopathogenic fungus was exposed, in polystyrene plates, to an artificial atmosphere containing a mixture of six VOCs formed by alcohols (ethanol, 3-methyl-1-butanol, 2-methyl-1-butanol and phenylethyl alcohol) and esters (ethyl acetate and ethyl octanoate), in the proportions found in the atmosphere naturally produced by yeast. Bean seeds artificially contamined with the pathogen were fumigated with the mixture of VOCs in sealed glass flasks for four and seven days. In the in vitro assays, the compounds 2-methyl-1-butanol and 3-methyl-1-butanol were the most active against S. sclerotiorum, completely inhibiting its mycelial growth at 0.8 µL mL-1, followed by the ethyl acetate, at 1.2 µL mL-1. Bean seeds fumigated with the VOCs at 3.5 µL mL-1 showed a 75% reduction in S. sclerotiorum incidence after four days of fumigation. The VOCs produced by S. cerevisiae have potential to control the pathogen in stored seeds.

Genetic control of potassium content of common bean seeds

The objective of this work was to investigate possible maternal effects on potassium content of common bean seeds, as well as to estimate the heritability and selection gains in early hybrid generations for this character and to evaluate the efficiency of genetic selection to improve the nutritional quality of common bean (Phaseolus vulgaris). Crosses with four cultivars from the Mesoamerican gene pool yielded the reciprocal F1 and F2 generations and the backcrossed populations (BCP1 and BCP2). The potassium content of the progenies was measured via nitric‑perchloric digestion and flame photometry. The potassium content in the tested progenies varied from 6.0 to 14.9 g kg-1 dry matter, and no significant maternal effect was observed. The narrow-sense heritability ranged from low (33.26%) to intermediate (43.05%). Partial dominance was observed for low potassium content in the seeds. No increase in potassium content was obtained through selection. Breeding common bean plants for increasing potassium content in seeds may be difficult because the local environment strongly influences the character.

Variability of ricin content in mature seeds of castor bean

The objective of this work was to evaluate ricin concentration in castor bean seeds (Ricinus communis) of 20 accessions from the Banco de Germoplasma de Mamoneira of the Embrapa Algodão, Campina Grande, PB, Brazil, using the Enzyme Linked Immunosorbent Assay. Significant differences were observed among accessions. BRA 3271 had the highest ricin concentration in seeds (32.18 ng µg-1), and BRS Paraguaçu had the lowest (3.53 ng µg-1). There is the possibility of selecting genotypes with different ricin concentrations, which can be used according on the interest of the breeding programs.