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.     

Evaluation of functional diversity in rhizobacterial taxa of a wild grass (Saccharum ravennae) colonizing abandoned fly ash dumps in Delhi urban ecosystem

Metal rich fly ash dumps may serve as repository of ecologically useful multi-functional rhizobacteria having potential use in the development of vegetation at the dumps. Therefore, in the present study bacteria from the rhizosphere of a wild perennial grass colonizing Indraprastha and Badarpur fly ash dumps of Delhi region were purified, identified and functionally characterized. The fly ash had low levels of nutrients, moisture and organic matter coupled with toxic levels of heavy metals (As, Cd, Cr, Cu, Hg, Ni, Pb and Zn). Both the dumps were mostly barren except for a few patches of Saccharum ravennae and some weedy species. Sixty five dominant, morphologically distinct rhizobacteria were purified, which belonged to 18 genera and 38 species. Gram-positive bacteria were dominating in the fly ash environment. Bacillus spp. and Paenibacillus spp. were common at both the dumps. Multi-metal tolerance was shown by diverse bacterial taxa. The minimum inhibitory concentration (MIC) was highest for As (12.5-20.0 mM) and Pb (7.5-10.0 mM), although many rhizobacteria also possessed significant tolerance to Cr, Zn, Ni, Cu, Co and Cd. The tolerance profiles of rhizobacteria to different metals may be ranked in the decreasing order as As > Pb > Cr > Zn > Ni > Cu > Co > Cd > Hg. Majority of rhizobacteria showed good siderophore activity. Multiple-metal tolerance was also coupled with high siderophore production in some of the isolates (Microbacterium barkeri IPSr74, Serratia marcescens IPSr90 and IPSr82, Enterococcus casseliflavus BPSr32, Bacillus sp. IPSr80, Pseudomonas aeruginosa BPSr43 and Brochothrix campestris BPSr3). Most of the bacteria could grow on nitrogen-deficient medium. However, the dominant nitrogen-fixers reported from the rhizosphere of other Saccharum species were not detected. S. marcescens IPSr90 was the only rhizobacterium, which showed ACC-deaminase (ACCD) activity. Proportion of phosphate-solubilizing bacteria was high. Considerable improvement in the seedling establishment, plant weight and shoot length in rhizobacterial inoculated plants of S. ravennae in fly ash environment indicated the significance of rhizobacteria in its colonization and spread to the dumps. Representative rhizobacteria, with high MIC (for most of the metals) and good plant growth promoting (PGP) traits comparable to commercially useful bacterial inoculants were identified as S. marcescens IPSr82 and IPSr90, P. aeruginosa BPSr43, Paenibacillus larvae BPSr106, Arthrobacter ureafaciens BPSr55, Paenibacillus azotofixans BPSr107 and E. casseliflavus BPSr32. S. ravennae and some of these rhizobacteria may be potentially useful for the development of inoculation technologies for conversion of barren fly ash dumps into ecologically and economically productive habitats.     

Multi-scale prediction of landscape resistance for tiger dispersal in central India

Context

Connectivity models for animal movement frequently use resistance surfaces, but rarely incorporate actual movement data and multiple scale drivers of landscape resistance.

Objectives

Using GPS data, we developed a multi-scale model of landscape resistance for tiger (Panthera tigris) dispersal in central India and evaluated the performance, interpretation and predictions against single scale models.

Methods

Six dispersing tiger paths were subjected to a path level analysis with conditional logistic regression to parameterize a resistance surface. We evaluated for 21 scales of available habitat and selected the best scale for each variable. We derived a scale-optimized multivariate path selection function and predicted landscape resistance across the landscape.

Results

The tigers preferred to move along areas with forest cover at relatively high elevations along the ridges with rugged topography at broad scale, while avoiding areas with agriculture-village matrix at fine scale. We found that the scale that was most supported by Akaike’s information criterion was not always the scale that maximized the magnitude (effect size) of the relationship. Further, the multi-scale optimized model differed substantially from the single scale models in terms of variable importance, magnitude of coefficients and predictions of connectivity.

Conclusions

Our results demonstrate that the variables in landscape resistance models produce markedly different predictions of population connectivity depending on the scales of analyses and interpretation. Thus, scale optimization in parameterization is critical for appropriate inferences and sound management strategies.

     

Identification and characterization of sources for photo‐ and thermo‐insensitivity in Vigna species

Cultivation of the same varieties of mungbean and blackgram across different seasons and locations is constrained by their photo‐ and thermo‐sensitive behaviour. Developing insensitive genotypes, which can fit well across all seasons, requires robust donors which would provide genes imparting this trait. This study was undertaken to identify such donors in the Vigna species. Forty‐eight accessions belonging to 13 Vigna species and eight released cultivars were evaluated under natural field conditions. Among these, two accessions, viz. V. umbellata (IC251442) and V. glabrescens (IC251372) were found photo‐ and thermo‐insensitive as these were able to flower and set pods at temperatures as high as 43.9°C and as low as 2.7°C. Pollen viability studies indicated viable pollen (>75% at 2.7°C and >85% at 41.9°C) and normal pollen tube growth at both the extremes of temperature. The identified V. glabrescens accession has long, constricted pods and dark green, mottled seeds while V. umbellata has smooth, curved pods and shining, oval, large seeds. Both these accessions can be utilized in developing photo–thermo insensitive genotypes in cultivated Vigna species.     

Effect of irradiated Ascaridia galli eggs on growth and cell-mediated immune responses in chickens

Growth and cell-mediated immune (CMI) responses were studied in 7-day-old chicks given orally 1000 irradiated (12.5 kR) or normal infective eggs of Ascaridia galli. Chicks immunised with irradiated eggs showed normal weight gains. CMI responses, as assessed by dinitrofluorobenzene (DNFB)-induced contact and delayed hypersensitivity reactions, were enhanced in the immunised group as compared with healthy controls, suggesting stimulation of CMI responses due to irradiation of A. galli eggs. CMI as well as growth responses were, however, found to be depressed in the birds administered normal infective eggs of A. galli. The present study highlights the role of the CMI response in protection against A. galli infection.     

Cross‐genera amplification of informative microsatellite markers from common bean and scarlet runner bean for assessment of genetic diversity in mungbean (Vigna radiata)

Mungbean (V. radiata) is an important Asiatic legume supplying inexpensive protein to a vast majority of vegetarian masses. To increase markers repertoire in mungbean, a study was conducted to analyse 384 microsatellite markers derived from common bean, scarlet runner bean and adzuki bean for their transferability and polymorphism. The results showed that 87 (24.71%) primer pairs could amplify DNA loci of 20 mungbean genotypes including one accession of V. trilobata, while 52 showed reliable banding and polymorphism. These showed different degrees of variability at each locus producing 250 alleles with the number of alleles varying from 2 to 9. The major allele frequency varied from 0.17 to 0.95, while the polymorphic information content of SSRs ranged between 0.09 and 0.86 with an average of 0.60 ± 0.16. UPGMA revealed three major clusters accommodating ~95% of the accessions while one accession of V. trilobata (‘NSB‐007’) did not group with any other genotype describing the discriminating power of informative microsatellites. This study identified a set of useful microsatellite markers to accelerate the genetic studies and breeding programme of mungbean.     

Momordica charantia L. (Cucurbitaceae) floral volatiles causing attraction of Epilachna dodecastigma (Coleoptera: Coccinellidae)

Epilachna dodecastigma (Wied.) (Coleoptera: Coccinellidae) causes economic losses to bitter gourd, Momordica charantia L. (Cucurbitaceae) production in India and Bangladesh because adults feed on the leaves and flowers resulting death of the plant. This insect is currently controlled by insecticides, which are harmful to human health and environment. We studied the behavioral responses of E. dodecastigma females to floral volatiles and synthetic compounds comparable to floral volatiles in a Y-tube olfactometer to determine their potential for monitoring this pest. The gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detector (GC-FID) analyses of M. charantia floral volatiles revealed the presence of 20 compounds. Myrcene was predominant followed by methyl jasmonate and 1-octadecanol in floral volatiles. Epilachna dodecastigma females were attracted to floral volatiles by a Y-tube olfactometer bioassay. Females were attracted to individual synthetic 1-heptanol, sabinene, myrcene and ocimene at 16, 1, 20, and 8 µg/25 mL methylene chloride in a dose response bioassay, respectively, and hence, these compounds might be used for insect pest management strategies such as baited traps.