Dynamic adsorption of α-pinene and linalool on silica nanoparticles for enhanced antifeedant activity against agricultural pests

The terpene compounds, α-pinene and linalool were formulated with silica nanoparticles (SNPs) by a simple immersion method and the antifeedant activity of these formulations was evaluated against two major agricultural pests, the tobacco cutworm (Spodoptera litura F.) and the castor semilooper (Achaea janata L.) in laboratory bioassays. The interaction between terpenes and SNPs, shelf-life, suspension stability as well as the bioactivity of the nanoformulations were also studied. Both these terpenes in their pure form are known to deter feeding in several lepidopterous insects. However, formulating these pure terpene compounds with nano silica enhanced their biological activity up to 25 times to S. litura. Fourier transform infrared spectroscopy analysis of these nano botanical formulations indicated the presence of hydrogen bonding between terpenes and the surface of SNPs. Transmission electron microscopy images confirmed the higher aggregation property of the SNPs. Suspension studies validated the improved shelf-life of the nano-biocomposites, while X-ray diffraction (XRD) revealed the amorphous nature of formulations and no crystalline impurities. Adsorption of α-pinene and linalool onto SNPs resulted in an effective formulation that enhanced the antifeedant potential of the individual terpenes against insects while producing longer shelf-life for the terpenes.     

Identification of molecular markers associated with sugar-related traits in a <Emphasis Type="Italic">Saccharum</Emphasis> interspecific cross

The cultivated sugarcane (Saccharum spp. hybrids, 2n = 100–130) is one crop for which interspecific hybridization involving wild germplasm has provided a major breakthrough in its improvement. Few clones were used in the initial hybridization event leading to a narrow genetic base for continued cultivar development. Molecular breeding would facilitate the identification and introgression of novel alleles/genes from the wild germplasm into cultivated sugarcane. We report the identification of molecular markers associated with sugar-related traits using an F₁ population derived from a cross between S. officinarum ‘Louisiana Striped’ × S. spontaneum ‘SES 147B’, the two major progenitor species of cultivated sugarcane. Genetic linkage maps of the S. officinarum and S. spontaneum parents were produced using the AFLP, SRAP and TRAP molecular marker techniques. The mapping population was evaluated for sugar-related traits namely, Brix (B) and pol (P) at the early (E) and late (L) plant growing season in the plant cane (04) and first ratoon (05) crops (04EB, 04LB, 04LP, 05EB and 05EP). For S. officinarum, combined across all the traits, a total of 30 putative QTLs was observed with LOD scores ranging from 2.51 to 7.48. The phenotypic variation (adj. R²) explained by all QTLs per trait ranged from 22.1% (04LP) to 48.4% (04EB). For S. spontaneum, a total of 11 putative QTLs was observed with LOD scores ranging from 2.62 to 4.70 and adj. R² ranging from 9.3% (04LP) to 43.0% (04LB). Nine digenic interactions (iQTL) were observed in S. officinarum whereas only three were observed in S. spontaneum. About half of the QTLs contributed by both progenitor species were associated with effects on the trait that was contrary to expectations based on the phenotype of the parent contributing the allele. Quantitative trait loci and their associated effects were consistent across crop-years and growing seasons with very few QTLs being unique to the early season. When the data were reanalyzed using the non-parametric discriminant analysis (DA) approach, significant marker-trait associations were detected for markers that were either identical to or in the vicinity of markers previously identified using the traditional QTL approach. Discriminant analysis also pointed to previously unidentified markers some of which remained unlinked on the map. These preliminary results suggest that DA could be used as a complementary approach to traditional QTL analysis in a crop like sugarcane for which saturated linkage maps are unavailable or difficult to obtain.     

Linkage mapping and genome analysis in a Saccharum interspecific cross using AFLP, SRAP and TRAP markers

Framework genetic linkage maps of two progenitor species of cultivated sugarcane, Saccharum officinarum ‘La Striped’ (2n = 80) and S. spontaneum ‘SES 147B’ (2n = 64) were constructed using amplified fragment length polymorphism (AFLP), sequence related amplified polymorphism (SRAP), and target region amplification polymorphism (TRAP) markers. The mapping population was comprised of 100 F₁ progeny derived from the interspecific cross. A total of 344 polymorphic markers were generated from the female (S. officinarum) parent, out of which 247 (72%) were single-dose (segregating in a 1:1 ratio) and 33 (9%) were double-dose (segregating in a 3.3:1 ratio) markers. Sixty-four (19%) markers deviated from Mendelian segregation ratios. In the S. spontaneum genome, out of a total of 306 markers, 221 (72%) were single-dose, 43 (14%) were double-dose, and 42 markers (14%) deviated from Mendelian segregation ratios. Linkage maps with Kosambi map distances were constructed using a LOD score ≥5.0 and a recombination threshold of 0.45. In Saccharum officinarum, 146 markers were linked to form 49 linkage groups (LG) spanning 1732 cM whereas, in S. spontaneum, 121 markers were linked to form 45 LG spanning 1491 cM. The estimated genome size of S. officinarum ‘La Striped’ was 2448 cM whereas that of S. spontaneum ‘SES 147B’ was 3232 cM. Based on the two maps, genome coverage was 69% in S. officinarum and 46% in S. spontaneum. The S. officinarum parent ‘La Striped’ behaved like an auto-allopolyploid whereas S. spontaneum ‘SES 147B’ behaved like a true autopolyploid. Although a large disparity exists between the two genomes, the existence of simple duplex markers, which are heterozygous in both parents and segregate 3:1 in the progeny, indicates that pairing and recombination can occur between the two genomes. The study also revealed that, compared with AFLP, the SRAP and TRAP markers appear less effective at generating a large number of genome-wide markers for linkage mapping in sugarcane. However, SRAP and TRAP markers can be useful for QTL mapping because of their ability to target gene-rich regions of the genome, which is a focus of our future research.     

Regeneration through somatic embryogenesis from petal-derived calli of Rosa hybrida L. cv arizona (hybrid tea)

Summary Somatic embryogenesis in callus cultures of petal explants of rose cv arizona is reported here. The calli from petals initiated on dicamba containing medium were friable and gave rise to embryos after several subcultures while those obtained from other explants did not show embryogenesis. Abscisic acid and phloroglucinol were necessary during maturation and plant development, respectively. The individual embryos grew into true-to-type plants.     

Quantification,Morphology and Ultrastructure of Preantral Follicles of Buffalo (Bubalus bubalis) Foetuses

The objective of this study was to determine the number, morphology and ultrastructure of preantral ovarian follicles of buffalo (Bubalus bubalis) foetuses at different ages. Quantification revealed number of primordial, primary and secondary follicles of 48 857 ± 17 506, 26 000 ± 20 452, 18 428 ± 10 875 and 18 375 ± 19 690, 225 ± 349, 326 ± 288 at 12–34 cm and 35–60 cm crown rump length (CRL), respectively. Follicular diameter values were 28.9 (±3.4), 34.7 (±5.9) and 59.4 (±12.6) μm; oocyte diameters were 21.7 (±2.8), 24.3 (±3.4) and 33.0 (±7.7) μm, and the numbers of follicular cells in the follicle equatorial section were 7.1 (±1.4), 12.0 (±2.4) and 13.8 (±2.4) for primordial, primary and secondary follicles, respectively. The primordial follicle consisted of an oocyte surrounded by a layer of flattened follicular cells with a normally eccentric oocyte nucleus. Dispersed Golgi complex, smooth endoplasmic reticulum, rounded mitochondria and several lipid vesicles were observed in the cytoplasm and cell junctions between the follicle cell membranes and the oocyte. This work describes the number, morphometry and ultrastructure of preantral follicles of buffalo foetuses, concluding that folliculogenesis is established between 8 and 34 cm CRL and that follicle number varies individually and according to age and that further studies are needed in this species.     

Competitiveness of native Bradyrhizobium japonicum strains in two different soil types

Interstrain competitiveness is a key factor affecting the performance of rhizobium inoculant. In the present study five native strains of Bradyrhizobium japonicum, namely SSF 4, SSF 5, SSF 6, SSF 7 and SSF 8, were assessed for their competitiveness in nodulating soybean using serological methods. The strains were inoculated individually or with the type strain USDA 110 at a 1:1 ratio. Nodule occupancy determined by immunofluorescence and dot immunoblot assay revealed that under in vitro conditions SSF 8 is more competitive than USDA 110 whereas the others were less competitive. The competitive ability of these strains was also estimated in pot culture in the field. In red soil both SSF 8 and USDA 110 were equally competitive whereas in black soil SSF 8 competed better than USDA 110 and produced more nodules. In a black soil field trial using a randomized block design, USDA 110 or SSF 8, when inoculated alone, occupied the majority of the nodules and enhanced nodule dry weight and shoot biomass. SSF 8 was more competitive when the strains were co-inoculated. Received: 1 November 1996     

Detection of Follicular Apoptosis in Water Buffalo (Bubalus bubalis) Ovary by Histology and Nick End Labelling Technique

The objective of this experiment was to assess the features and extent of follicular apoptosis in the water buffalo (Bubalus bubalis) ovary using classical histology and nick end labelling technique. Ovaries (n = 40) procured from the slaughterhouse were used for the study. The sections (5 μm) were used for detection of terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labelling (TUNEL) and classical histology (H&E). Those follicles showing ≥ 5% TUNEL positivity (TUNEL assay) and pyknotic nuclei (histology) in granulosa cells were classified as atretic. Based on histology, the atretic primary and secondary follicles (%) were 93.82 and 95.62 respectively. The histology study reveals that the rates (%) of atresia in <1, 1–3, 3–5 mm and >5 mm were 36.90, 40.50, 62.84 and 74.5 respectively. Further the atretic tertiary follicles (%) were significantly lower than the primary and secondary classes of follicles. TUNEL assay reveals that the atretic rate (%) of tertiary follicles in <1, 1–3, 3–5 and ≥ 5 mm class follicles were 50.88, 53.84, 81.81 and 36.36 respectively. The percentage of atresia in >5 mm diameter follicles were significantly lower in TUNEL than histology. Percentages of granulosa and thecal cells positive for atresia by TUNEL were 30.7 ± 0.53 and 13.82 ± 0.18 respectively per follicle. The initial structural changes in atretic follicles were seen primarily in the granulosa cells. In severely atretic follicles TUNEL positive granulosa cells along with theca cells have to be considered in assessing the rate and extent of atresia.