An in vitro propagation system was developed for castor-bean (Ricinus communis L. cv. TMV 6) through cotyledon derived callus cultures. The impact of different concentrations of auxins, cytokinins, additives, amino acids and sugars were evaluated for callus induction and shoot proliferation. Green compact nodular organogenic callus was obtained on the medium fortified with Murashige and Skoog (MS) salts, B5 vitamins, 2.0 mg dm -3 6-benzyladenine and 0.8 mg dm -3 α-naphthalene acetic acid (NAA). Multiple shoot proliferation from the callus cultures was achieved on the medium with MS salts, B5 vitamins, 2.5 mg dm -3 thidiazuron (TDZ), 0.4 mg dm -3 NAA and 15 mg dm -3 glutamine. During multiple shoot induction the phenolic secretion was controlled by the addition of 15 mg dm -3 polyvinylpyrolidone. The proliferated shoots were elongated on the medium comprising MS salts, B5 vitamins, 1.5 mg dm -3 TDZ and 0.3 mg dm -3 gibberellic acid. The elongated shoots were rooted on the medium containing MS salts, B5 vitamins, 0.3 mg dm -3 indole-3-butyric acid and 0.6 mg dm -3 silver nitrate. After root induction, the plants were hardened in earthen pots containing sand, soil and vermiculite.

Problem statement: The present investigation described a simple and reproducible protocol for transgenic cotton regeneration and characterization of chitinase (Chi II) gene expression against two different fungal pathogens in cotton. Approach: Transgenic cotton (Gossypium hirsutum cv. SVPR2) plants were produced by pCambia-bar-Chi II (13.8 kb) under the control of the CaMV 35S promoter, harbored in the strain LBA 4404 Agrobacterium tumefaciens by using shoot tip explants. Results: Finally, from the 10 experiments, 21.8% of transformation frequency was recorded. Segregation ratio of 3:1 was recorded in the T 0 plant seeds. Polymerase chain reaction and southern blotting analysis were used to confirm the integration of Chi II transgene in the T 0 plants genome of putative transgenics. Quantitiave and qualitative (SDS-PAGE) analyses were also carried out to confirm the expression of chitinase enzyme in T 0 plants. Further, randomly selected transgenic plants (T 0 ) were analyzed for disease tolerance by evaluating them with spores of Fusarium oxysporum and Alternaria macrospora. All the selected PCR positive plants showed enhanced disease resistance against Fusarium wilt. The plants selected randomly showed an enhanced survival rate compared with the control when they were grown in earthen pots inoculated with 1×10 5 spores 100 −1 g of soil mixture.Another four randomly selected plantlets were sprayed with spores of Alternaria macrospora in order to test their tolerance to Alternaria leaf spot disease. After 20 days of culture, the number of lesions per leaf and the lesion length per leaf spot in non-transferred leaves increased. In the case of transgenic plantlets, lesion formation was completely absent. Conclusion: The disease resistance against Fusarium wilt and Alternaria leaf spot in cotton strains would serve as good breeding materials for producing fungal disease resistant cotton varieties.

Chromosome analyses on different species of Orchidaceae namely, Vanda alpine Lindl, Vanda amesiana Reichb.f. Vanda coerulea Griff, Vanda coerulescence Griff, Vanda cristata Lindl, Vanda densiflora Lindl and Vanda hookeriana Reichb. f.. Deviant report as against the previous records of chromosome numbers have been made in Vanda alpine Lindl, Vanda amesiana Reichb.f. Vanda coerulea Griff, Vanda coerulescence Griff, Vanda cristata Lindl, Vanda densiflora Lindl and Vanda hookeriana Reichb.f.a. In all other species studied the present report of chromosome numbers are in correlation with those of earlier reports. The first record of chromosome number has been made in 2 species namely Vanda coerulescence Griff 2n-42, Vanda cristata Lindl 2n–40 and Vanda densiflora Lindl 2n-42. Most of the species of Vanda alpine Lindl 2n-38, Vanda amesiana Reichb.f. 2n–38, Vanda hookeriana Reichb.f. 2n–38 and Vanda coerulea Griff 2n–42. Vanda cristata 2n–40, Vanda densiflora 2n-42. Chromosome should have been originated by aneuploidy. The study of karyotype analyses shows that each and every species has a distinct karyotype. Therefore, karyotype alternations of chromosomes may have also play important role in speciation along with aneuploidy.

The sesame seed protein was electrophoretically separated through sodium dodecylesulphate polyacrylamide gel electrophoresis (SDS-PAGE). Electrophoretic banding homology among the genotypes was established not only through the banding pattern but also through estimation of dissimilarity percentage in protein profile. The advantage of SDS–PAGE is that the proteins are separated by their molecular weight and it has become one of the most widely used technique for storage protein separation. Freshly harvested seeds of 21 advance lines and their 8 parental lines were taken for the experiment of our study. The genetic diversity and the relationships among the 29 genotypes were evaluated using sodium dodecylesulphate polyacrylamide gel electrophoresis (SDS –PAGE) which shows difference in number of bands, band width and intensity for different genotypes. It was possible to identify as many as 21 different bands and these can be easily visualized. Dendogram of 21 advance lines and their 8 parents based on SDS – PAGE data using dissimilarity percentage was presented here. Mainly two clusters obtained from the dendrogram for all the 29 genotypes where cluster A forms 3 and cluster B forms 6 sub- clusters. Clustering pattern, size and constituents on the basis of electrophoretic banding pattern was independent of the same through the D² analysis on the basis of biochemical parameters and presence of distinct phylogenetic relationship could be recognized among the genotypes. The dissimilarity % in protein profile indicated that the B- 9 vs. adv. line 5, B-9 vs. adv. line 8, R – 9 vs. adv. line 8, adv. Line 14 vs. adv.18, HT-1 vs. adv. 11 would be useful in identifying parents for heterosis breeding.