Fruit dehiscence mechanism and release of dimorphic seeds with different germination properties in Commelina erecta.

Commelina erecta is a successful weed species. The aims of this study were to analyse the morpho-anatomy of the fruit and dimorphic seeds of the weed C. erecta, the dynamics and type of dormancy, and water entry. Flowers and fruits at different development stages were processed using standard anatomical techniques. Besides, experiments of imbibition, germinability and water entry were performed on both seed types. In the fruit of C. erecta, free and coated seeds are developed within dehiscent and indehiscent carpels, respectively. Dehiscent carpels open through a region of mechanical weakness in the dorsal vascular bundle. This region does not form in the indehiscent carpel. The main anatomical differences between the two seed types were observed in the testa and in the number of covering layers. Imbibition experiments showed that the covering of both seed types is water permeable, so these seeds lack physical dormancy and may exhibit physiological dormancy. Germinability experiments showed that the dormancy in free seeds is variable throughout the reproductive season, whereas, in coated seeds, it is high throughout the reproductive season. The embryotega is an area where the hardness of the seed coat is interrupted and facilitates water entry. Differences in the morpho-anatomy of carpels result in the formation of dimorphic seeds with different covering layers and different germination properties. These different properties allow some seeds germinate immediately after falling from the mother plant, and others to be incorporated into the seed bank. These results are useful for designing weed management strategies in agroecosystems.

Distribution of Glyphosate-Resistance in Echinochloa crus-galli Across Agriculture Areas in the Iberian Peninsula.

The levels of resistance to glyphosate of 13 barnyard grass (Echinochloa crus-galli) populations harvested across different agriculture areas in the Southern Iberian Peninsula were determined in greenhouse and laboratory experiments. Shikimate accumulation fast screening separated the populations regarding resistance to glyphosate: susceptible (S) E2, E3, E4, and E6 and resistant (R) E1, E5, E7, E8, E9, E10, E11, E12, and E13. However, resistance factor (GR50 E1-E13/GR50 E6) values separated these populations into three groups: (S) E2, E3, E4, and E6, (R) E1, E5, E7, E8, and E9, and very resistant (VR) E10, E11, E12, and E13. 14C-glyphosate assays performed on two S populations (E2 and E6) showed greater absorption and translocation than those found for R (E7 and E9) and VR (E10 and E12) populations. No previous population metabolized glyphosate to amino methyl phosphonic acid (AMPA) and glyoxylate, except for the E10 population that metabolized 51% to non-toxic products. The VR populations showed two times more 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity without herbicide than the rest, while the inhibition of the EPSPS activity by 50% (I50) required much higher glyphosate in R and VR populations than in S populations. These results indicated that different target-site and non-target-site resistance mechanisms were implicated in the resistance to glyphosate in E. crus-galli. Our results conclude that resistance is independent of climate, type of crop, and geographic region and that the level of glyphosate resistance was mainly due to the selection pressure made by the herbicide on the different populations of E. crus-galli studied.

Low temperatures enhance the absorption and translocation of 14C-glyphosate in glyphosate-resistant Conyza sumatrensis.

Influence of low temperatures on the glyphosate efficacy was studied in glyphosate-resistant (R) and -susceptible (S) Conyza sumatrensis biotypes. For this purpose, the physiological and enzymatic aspects involved were characterized under two growing temperature regimes [high (30/20 °C) and low 15/5 °C temperatures day/night]. The R biotype was 5.5 times more resistant than the S biotype at high temperatures; however, this R-to-S ratio decreased to 1.6 at low temperatures. At 96 h after treatment (HAT), the shikimic acid accumulation was higher in the S biotype in both temperature regimes (4.6 and 1.9 more shikimic acid at high and low temperatures, respectively), but the accumulation of the R biotype increased 2.6 times at low temperatures compared to high ones. From 24 to 96 HAT, the 14C-glyphosate absorption ranged from 28 to 65% (percentage reached from 48 HAT) at low temperatures, and from 20 to 50% at high temperatures (gradual increase), but there were no differences between C. sumatrensis biotypes within each temperature regime. At high temperatures, the 14C-glyphosate translocation was different between biotypes, where the R one retained at least 10% more herbicide in the treated leaves than the S biotype at 96 HAT. So, the S biotype translocated 40% of 14C-glyphosate absorbed to roots, and the R biotype translocated only 28% of herbicide at the same period. At low temperatures, there were no differences between biotypes, and at 96 HAT, the 14C-glyphosate found in treated leaves was ˜47% and up to ˜42% reached the roots, i.e., the resistance mechanism was suppressed. The basal and enzymatic activities of the 5-enolpyruvyishikimate 3-phosphate synthase were different between temperature regimes, but there was no differences between biotypes within each temperature regime, showing that target-site resistance mechanisms did not contribute in the glyphosate resistance of the R biotype. Low temperatures enhanced the absorption and translocation of glyphosate by suppressing the resistance mechanisms improving its efficacy on resistant plants. This is the first characterization about the role of temperatures in the glyphosate efficacy on C. sumatrensis.

Cytosolic Glyceraldehyde-3-Phosphate Dehydrogenase Is Phosphorylated during Seed Development.

Cytosolic glyceraldehyde-3-phosphate dehydrogenase (NAD-GAPDH) is involved in a critical energetic step of glycolysis and also has many important functions besides its enzymatic activity. The recombinant wheat NAD-GAPDH was phosphorylated in vitro at Ser205 by a SNF1-Related protein kinase 1 (SnRK1) from wheat heterotrophic (but not from photosynthetic) tissues. The S205D mutant enzyme (mimicking the phosphorylated form) exhibited a significant decrease in activity but similar affinity toward substrates. Immunodetection and activity assays showed that NAD-GAPDH is phosphorylated in vivo, the enzyme depicting different activity, abundance and phosphorylation profiles during development of seeds that mainly accumulate starch (wheat) or lipids (castor oil seed). NAD-GAPDH activity gradually increases along wheat seed development, but protein levels and phosphorylation status exhibited slight changes. Conversely, in castor oil seed, the activity slightly increased and total protein levels do not significantly change in the first half of seed development but both abruptly decreased in the second part of development, when triacylglycerol synthesis and storage begin. Interestingly, phospho-NAD-GAPDH levels reached a maximum when the seed switch their metabolism to mainly support synthesis and accumulation of carbon reserves. After this point the castor oil seed NAD-GAPDH protein levels and activity highly decreased, and the protein stability assays showed that the protein would be degraded by the proteasome. The results presented herein suggest that phosphorylation of NAD-GAPDH during seed development would have impact on the partitioning of triose-phosphate between different metabolic pathways and cell compartments to support the specific carbon, energy and reducing equivalent demands during synthesis of storage products.

Glyphosate-induced structural variations in Commelina erecta L. (Commelinaceae).

The purpose of this study was to analyze the structural variations of Commelina erecta, a glyphosate-tolerant weed, produced by this herbicide. Regrowth shoots (vegetative cloning axes) and seedlings and two glyphosate concentrations: 900 and 1800 grams of acid equivalent per hectare (ga.e. ha⁻¹) were used. The following variations were observed: foliar shape and size changes, changes in the length of internodes, variations in the quantity of inflorescences, low seed production, and differences in the number of branches. After treatment, different phenotypic responses were observed as a result of differential sensitivity to glyphosate. Most of the vegetatively-propagated plants treated with 900 g a.e. ha⁻¹ revealed the same morphological structure as that of controls (i.e. three clearly identifiable zones: a basal or branching zone, a middle zone where branch production is inhibited, and an apical zone where only floral branches are produced). By contrast, the 900 ga.e. ha⁻¹ treated seedlings and the 1800 g a.e. ha⁻¹ treated regrowth shoots and seedlings showed two phenotypic responses, exhibiting differences in the quantity and location of branches and in the survival time of axes. The main variation is seen in branch production in the region that is usually inhibited. The different phenotypic responses and morphological changes suggest that C. erecta shows a very high intraspecific variability, which allows it to survive and thrive even in environments with intensive glyphosate application. These results underscore the need to solve the problem of tolerant weeds by means of an integrated approach including alternative management practices grounded on knowledge of the species and their behavior.