Genetic polymorphism detection in brazilian perennial cottons (Gossypium spp.) using an ISSR marker system and its application for molecular interspecific differentiation.

BACKGROUND: The semi-domesticated Brazilian perennial cotton (Gossypium spp.) germplasm is considered a source of variability for creating modern upland cotton varieties. Here we used Inter-simple Sequence Repeat (ISSR) markers to detect intra and interspecific genetic polymorphism in Gossypium hirsutum L. r. marie-galante and Gossypium barbadense L. and to use molecular data to assessing genetic diversity and molecular discrimination of these species. METHODS AND RESULTS: The sets contained 12 G. barbadense genotypes and 16 G. hirsutum genotypes from a Brazilian collection. The 11 ISSR primers were used for genotyping yielded 101 bands (polymorphism = 47.5%) and were classified as moderately informative (PIC = 0.304). The ISSR markers exposed a greater diversity in G. hirsutum (P = 24.72%; HE =0.071 and I = 0.111) as compared to G. barbadense (P = 17.98%, HE = 0.043 and I = 0.070). The AMOVA analysis showed that 89.47% of the genetic variation was partitioned within species which is supported by Nei's genetic differentiation (Gst = 0.598) and gene flow (Nm = 0.338), suggesting that strong reproductive barriers between species. The UPGMA Cluster Analysis, Principal Coordinate Analysis and Bayesian Model-Based Structural Analysis divided the 28 genotypes into two main clades consistent with the taxonomical delimitation. CONCLUSION: The ISSR marker system offers a new approach to determining molecular differences between two cotton species (G. hirsutum L. r. marie-galante and G. barbadense L.). This study can expand the molecular marker resources for the identification and improvement of our knowledge about the genetic diversity and relationships between perennial cotton genotypes.

Molecular and biochemical responses of sesame (Sesame indicum L.) to rhizobacteria inoculation under water deficit.

Introduction: Water scarcity is a challenge for sesame cultivation under rainfed conditions. In this scenario, a potential strategy to alleviate the water deficit is the application of plant growth-promoting bacteria. The objective of this study was to analyze the interaction of rhizobacteria with sesame cultivation under water deficit conditions. Methods: An experiment was conducted in pots in a greenhouse using the BRS Morena sesame cultivar. The experimental design was completely randomized in a factorial scheme: 2 (irrigation regimes - daily irrigation and water deficit by suspending irrigation until 90% stomatal closure) x 6 (treatments with nitrogen or inoculants), with 5 replications. The types of fertilization were characterized by the addition of nitrogen (ammonium sulfate; 21% N), inoculants based on Bacillus spp. (pant001, ESA 13, and ESA 402), Agrobacterium sp. (ESA 441), and without nitrogen (control). On the fifth day after the suspension of irrigation, plant material was collected for gene expression analysis (DREB1 and HDZ7), activities of antioxidant enzymes (superoxide dismutase and catalase), relative proline content, and photosynthetic pigments. At the end of the crop cycle (about 85 days), production characteristics (root dry matter, aboveground dry matter, number of capsules, and thousand seed weight), as well as leaf nitrogen (N) and phosphorus (P) content, were evaluated. Results and Discussion: There was a positive effect on both production and biochemical characteristics (proline, superoxide dismutase, catalase, and photosynthetic pigments). Regarding gene expression, most of the inoculated treatments exhibited increased expression of the DREB1 and HDZ7 genes. These biological indicators demonstrate the potential of rhizobacteria for application in sesame cultivation, providing nutritional supply and reducing the effects of water deficit.

Elite Bradyrhizobium strains boost biological nitrogen fixation and peanut yield in tropical drylands.

Peanut (Arachis hypogaea L.) is an important crop for the family-based systems in the tropics, mainly in Brazil. In the Brazilian drylands, peanuts are cropped in low technological systems, and cheap and efficient technologies are needed to improve crop yield and sustainability. Despite this importance, few data are available on selecting efficient peanut rhizobia in experiments under different edaphoclimatic conditions. This work evaluated the agronomic efficiency and the biological nitrogen fixation (BNF) by two elite Bradyrhizobium strains under four different fields in the Brazilian semiarid region. We compared a new efficient strain Bradyrhizobium sp. ESA 123 with the reference strain B. elkanii SEMIA 6144, currently used in peanut rhizobial inoculants in Brazil. Besides the inoculated treatments, two uninoculated controls were assessed (with and without 80 kg ha-1 of N-urea). The BNF was estimated by the δ15N approach in three out of four field assays. BNF contribution was improved by inoculation of both Bradyrhizobium strains, ranging from 42 to 51% in Petrolina and 43 to 60% in Nossa Senhora da Glória. Peanuts' yields benefited from the inoculation of both strains and N fertilization in all four assays. Nevertheless, the results showed the efficiency of both strains under different edaphoclimatic conditions, indicating the native strain ESA 123 as a potential bacterium for recommendation as inoculants for peanuts in Brazil, mainly in drylands.

Validating a probe from GhSERK1 gene for selection of cotton genotypes with somatic embryogenic capacity.

Substantial progress is being reported in the techniques for plant transformation, but successful regeneration of some genotypes remains a challenging step in the attempts to transform some recalcitrant species. GhSERK1 gene is involved on embryo formation, and its overexpression enhances the embryogenic competence. In this study we validate a short GhSERK1 probe in order to identify embryogenic cotton genotypes using RT-qPCR and blotting assays. Cotton genotypes with contrasting somatic embryogenic capacity were tested using in vitro procedures. High expression of transcripts was found in embryogenic genotypes, and the results were confirmed by the RT-PCR-blotting using a non-radioactive probe. The regeneration ability was confirmed in embryogenic genotypes. We confirmed that GhSERK1 can be used as marker for estimating the somatic embryogenesis ability of cotton plants.

Plant promoters: an approach of structure and function.

With current advances in genomics, several technological processes have been generated, resulting in improvement in different segments of molecular research involving prokaryotic and eukaryotic systems. A widely used contribution is the identification of new genes and their functions, which has led to the elucidation of several issues concerning cell regulation and interactions. For this, increase in the knowledge generated from the identification of promoters becomes considerably relevant, especially considering that to generate new technological processes, such as genetically modified organisms, the availability of promoters that regulate the expression of new genes is still limited. Considering that this issue is essential for biotechnologists, this paper presents an updated review of promoters, from their structure to expression, and focuses on the knowledge already available in eukaryotic systems. Information on current promoters and methodologies available for studying their expression are also reported.

Effect of a Bowman-Birk proteinase inhibitor from Phaseolus coccineus on Hypothenemus hampei gut proteinases in vitro.

The coffee berry borer, Hypothenemus hampei (Ferrari), is an important devastating coffee pest worldwide. Both trypsin and chymotrypsin enzyme activities from H. hampei larval midgut can be inactivated by proteinaceous enzyme-inhibitors. A serine proteinase inhibitor belonging to the Bowman-Birk class was purified from a wild accession of Phaseolus coccineus L. seeds. The inhibitor (PcBBI1) is a cysteine-rich protein that is heat-stable at alkaline pH. MALDI-TOF/MS analysis showed that PcBBI1 occurs in seeds as a monomer (8689 Da) or dimer (17,378 Da). Using in vitro inhibition assays, it was found that PcBBI1 has a high inhibitory activity against H. hampei trypsin-like enzymes, bovine pancreatic chymotrypsin, and trypsin. According to this, PcBBI1 could be a promising tool to make genetically modified coffee with resistance to coffee berry borer.