Potential,constraints and applications of in vitro methods in improving grain legumes

Grain legumes, the important constituents of sustainability‐based cropping systems and energy‐limited vegetarian diets have long been the subject of scientific research. Tremendous technological strides were made in the so‐called orphan crops, in terms of both varietal improvement and generation of basic information. Despite recalcitrancy and high genotype dependency, in vitro culture techniques such as organogenesis, in vitro mutagenesis, embryo rescue and in vitro gene transfer have been deployed for improvement of several grain legumes and these played an important role in introgression of desirable genes from related and distant species and creation of additional genetic variability. Stable and reproducible regeneration protocols resulted in the development of genetically modified chickpea, pigeon pea, cowpea, mungbean, etc., while embryo rescue was deployed successfully for recovery of interspecific recombinants, a few of them exploited for the development of commercial cultivars. Nevertheless, doubled haploidy witnessed limited success and protoplast regeneration and in vitro mutagenesis remained of academic interest. The present review focuses on the progress, achievements, constraints and perspectives of using in vitro technology in grain legume improvement.     

Inter-site transferability of crop varieties: another approach for analyzing multi-locational variety trials

Summary Improved crop varieties developed at experimental stations are intended for release for production in farmers' fields which are not necessarily the test sites. Therefore, it is important to measure the inter-site transferability of the crop variety. This study was undertaken to establish a statistical measure of inter-site transferability of a variety. Data on grain yield from two sets of multi-locational trials on barley and four sets of the wheat trials conducted by the Cereal Program, International Center for Agricultural Research in the Dry Areas, Syria, were analyzed to evaluate the inter-site transferability of the varieties. Genotype × environment interaction was significant and experimental errors were heterogeneous in each set of trials. Correlations between the slope (of linear regression of variety mean on location mean) and variety mean were significant (P < 0.01) in all trials. Correlations between the developed inter-site transferability statistic and the variety mean were significant (P < 0.05) in only two trials. Correlations between mean yield and probability of the inter-site transferability statistic were insignificant in all trials. The inter-site transferability statistic and the associated probability of the transferability may be used to select varieties which are high yielding as well as transferable to a new environment. The strong association of slope with mean yield restricted the scope of varietal selection but the introduced statistic did not suffer from this drawback.     

Cover crop effect on soil carbon fractions under conservation tillage cotton

Cover crops may influence soil carbon (C) sequestration and microbial biomass and activities by providing additional residue C to soil. We examined the influence of legume [crimson clover (Trifolium incarnatum L.)], nonlegume [rye (Secale cereale L.)], blend [a mixture of legumes containing balansa clover (Trifolium michelianum Savi), hairy vetch (Vicia villosa Roth), and crimson clover], and rye + blend mixture cover crops on soil C fractions at the 0–150 mm depth from 2001 to 2003. Active fractions of soil C included potential C mineralization (PCM) and microbial biomass C (MBC) and slow fraction as soil organic C (SOC). Experiments were conducted in Dothan sandy loam (fine-loamy, kaolinitic, thermic, Plinthic Kandiudults) under dryland cotton (Gossypium hirsutum L.) in central Georgia and in Tifton loamy sand (fine-loamy, siliceous, thermic, Plinthic Kandiudults) under irrigated cotton in southern Georgia, USA. Both dryland and irrigated cotton were planted in strip tillage system where planting rows were tilled, thereby leaving the areas between rows untilled. Total aboveground cover crop and cotton C in dryland and irrigated conditions were 0.72–2.90 Mg C ha⁻¹ greater in rye + blend than in other cover crops in 2001 but was 1.15–2.24 Mg C ha⁻¹ greater in rye than in blend and rye + blend in 2002. In dryland cotton, PCM at 50–150 mm was greater in June 2001 and 2002 than in January 2003 but MBC at 0–150 mm was greater in January 2003 than in June 2001. In irrigated cotton, SOC at 0–150 mm was greater with rye + blend than with crimson clover and at 0–50 mm was greater in March than in December 2002. The PCM at 0–50 and 0–150 mm was greater with blend and crimson clover than with rye in April 2001 and was greater with crimson clover than with rye and rye + blend in March 2002. The MBC at 0–50 mm was greater with rye than with blend and crimson clover in April 2001 and was greater with rye, blend, and rye + blend than with crimson clover in March 2002. As a result, PCM decreased by 21–24 g CO₂–C ha⁻¹ d⁻¹ but MBC increased by 90–224 g CO₂–C ha⁻¹ d⁻¹ from June 2001 to January 2003 in dryland cotton. In irrigated cotton, SOC decreased by 0.1–1.1 kg C ha⁻¹ d⁻¹, and PCM decreased by 10 g CO₂–C ha⁻¹ d⁻¹ with rye to 79 g CO₂–C ha⁻¹ d⁻¹ with blend, but MBC increased by 13 g CO₂–C ha⁻¹ d⁻¹ with blend to 120 g CO₂–C ha⁻¹ d⁻¹ with crimson clover from April 2001 to December 2002. Soil active C fractions varied between seasons due to differences in temperature, water content, and substrate availability in dryland cotton, regardless of cover crops. In irrigated cotton, increase in crop C input with legume + nonlegume treatment increased soil C storage and microbial biomass but lower C/N ratio of legume cover crops increased C mineralization and microbial activities in the spring.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of safflower and the efficient recovery of transgenic plants via grafting

Background

Safflower (Carthamus tinctorius L.) is a difficult crop to genetically transform being susceptible to hyperhydration and poor in vitro root formation. In addition to traditional uses safflower has recently emerged as a broadacre platform for the production of transgenic products including modified oils and pharmaceutically active proteins. Despite commercial activities based on the genetic modification of safflower, there is no method available in the public domain describing the transformation of safflower that generates transformed T₁ progeny.

Results

An efficient and reproducible protocol has been developed with a transformation efficiency of 4.8% and 3.1% for S-317 (high oleic acid content) and WT (high linoleic acid content) genotypes respectively. An improved safflower transformation T-DNA vector was developed, including a secreted GFP to allow non-destructive assessment of transgenic shoots. Hyperhydration and necrosis of Agrobacterium-infected cotyledons was effectively controlled by using iota-carrageenan, L-cysteine and ascorbic acid. To overcome poor in vitro root formation for the first time a grafting method was developed for safflower in which ~50% of transgenic shoots develop into mature plants bearing viable transgenic T₁ seed. The integration and expression of secreted GFP and hygromycin genes were confirmed by PCR, Southern and Western blot analysis. Southern blot analysis in nine independent lines indicated that 1-7 transgenes were inserted per line and T₁ progeny displayed Mendelian inheritance.

Conclusions

This protocol demonstrates significant improvements in both the efficiency and ease of use over existing safflower transformation protocols. This is the first complete method of genetic transformation of safflower that generates stably-transformed plants and progeny, allowing this crop to benefit from modern molecular applications.

     

The immune response to anesthesia: Part 1

ObjectiveTo review the immune response to anesthesia including mechanical ventilation, inhaled anesthetic gases, and injectable anesthetics and sedatives.Study designReview.Methods and databasesMultiple literature searches were performed using PubMed and Google Scholar from spring 2012 through fall 2013. Relevant anesthetic and immune terms were used to search databases without year published or species constraints. The online database for Veterinary Anaesthesia and Analgesia and the Journal of Veterinary Emergency and Critical Care were searched by issue starting in 2000 for relevant articles.ConclusionRecent research data indicate that commonly used volatile anesthetic agents, such as isoflurane and sevoflurane, may have a protective effect on vital organs. With the lung as the target organ, protection using an appropriate anesthetic protocol may be possible during direct pulmonary insults, including mechanical ventilation, and during systemic disease processes, such as endotoxemia, generalized sepsis, and ischemia-reperfusion injury.     

Exploring genetic diversity for heat tolerance among lentil (Lens culinaris Medik.) genotypes of variant habitats by simple sequence repeat markers

The study was carried out to assess genetic diversity among 119 lentil genotypes grown in different habitats for heat tolerance using morpho‐physiological and reproductive traits and SSR markers. High‐temperature stress was applied at seedling (35/33°C) and anthesis stages (35/20°C) to study the effects on morpho‐physiological and reproductive traits under hydroponic condition, which was compared with non‐stressed and stressed field conditions. A set of 209 alleles were identified by 35 SSR markers among the genotypes. Genetic diversity and polymorphism information content values varied between 0.0494–0.859 and 0.0488–0.844, with mean values of 0.606 and 0.563, respectively. Genotypes were clustered into nine groups based on SSR markers. Morpho‐physiological and reproductive traits under heat stress were found to be significantly different among SSR clusters. These findings suggest that heat adaptation is variable among the genotypes and the tolerant materials can be evolved through hybridization using parents from different clusters with diverse mechanisms of heat tolerance.     

Studies on pesticides based on coumarin. II. Synthesis and antifungal activity of ten 2-alkyl-7, 8-dihydro-3-hydroxynaphtho[1,2-c]chromen-6-ones

A series of 2-alkyl-7, 8-dihydro-3-hydroxynaphtho[1,2-c]chromen-6-ones was synthesised by the condensation of ethyl 3, 4-dihydro-1-oxonaphthalene-2-carboxylate with substituted phenols in the presence of POCl₃. The compounds were characterised and tested for their toxicity towards the mycelial growth of seven phytopathogenic fungi in culture. Drechslera oryzae, Rhizoctonia solani and Colletotrichum falcatum exhibited maximum sensitivity to these compounds whereas Macro-phomina phaseolina, Fusarium solani, Alternaria alternata and Pythium aphanidermatum were less sensitive. 2-Ethyl-7, 8-dihydro-3-hydroxy-naphtho[1,2-c]chromen-6-ones possessed greatest toxicity with EC₅₀ values ranging from 0.2 to 2.5 μg ml^?1 against all fungi except A. alternata and P. aphanidermatum.     

Interaction Between Sorghum Protein Extraction and Precipitation Conditions on Yield,Purity, and Composition of Purified Protein Fractions

Sorghum proteins have the potential to be used as a bio‐industrial renewable resource for applications such as biodegradable films and packaging. This project was designed to evaluate the effect of interactions between sorghum protein extraction and precipitation conditions on the yield, purity, and composition of sorghum protein fractions. Proteins were extracted with 70% ethanol under nonreducing conditions, with ultrasound, or under reducing conditions using either sodium metabisulfite or glutathione as the reducing agent. Several conditions were used to isolate the extracted proteins through precipitation, including lowering ethanol concentrations alone or in combination with lowering to pH 2.5, or by adding 1M NaCl to the extract. Combinations of these conditions were also tested. All precipitation conditions effectively precipitated proteins and lowering the pH and adding 1M NaCl to the extracts enhanced precipitation in some cases. However, the conditions that precipitated the maxium amount of protein or highest purity of protein varied according to how the proteins were initially extracted. Precipitated proteins were characterized by RP‐HPLC, SEC, HPCE, and SDS‐PAGE to compare the protein fractions composition. Nonreduced and sonicated samples had a much wider M_w distribution than reduced extracts. Thus, extraction and precipitation conditions influenced the isolated proteins yield, purity, and composition. Because the extraction and purification processes influenced the composition, purity, and biochemical properties, it may be possible to prepare protein fractions with unique functionalities for specific end‐uses.