Satyesh Chandra Roy

Genetic stability of three economically important micropropagated banana (Musa spp.) cultivars of lower Indo-Gangetic plains, as assessed by RAPD and ISSR markers

An efficient micropropagation protocol produced large number of plants of the three elite banana (Musa spp.) cultivars Robusta (AAA), Giant Governor (AAA) and Martaman (AAB) from shoot tip meristem. The genetic relationships and fidelity among the cultivars and micropropagated plants as assessed by random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers, revealed three somaclonal variants from Robusta and three from Giant Governor. A total of 5330 RAPD and 2741 ISSR fragments were generated with 21 RAPD and 12 ISSR primers in micropropagated plants. The percentage of polymorphic loci by RAPD and ISSR were found to be 1.75, 5.08 in Robusta and 0.83, 5.0 in Giant Governor respectively. Among the two marker systems used, ISSR fingerprinting detected more polymorphism than RAPD in Robusta and Giant Governor with most of the primers showing similar fingerprinting profile, whereas Martaman revealed complete genetic stability.

In vitro regeneration and micropropagation of Aloe vera L.

Abstract Rapid propagation by the formation of shoots from calli of Aloe vera was obtained in the present investigation. Callus formation was induced in stem segments from young axillary shoots grown on the underground rhizomatous stem. The use of polyvinylpyrrolidone (PVP) in the nutrient media reduced the secretion of phenolic substances from the explant. Murashige and Skoogs basal medium containing 1 mg l−1 2,4-dichlorophenoxyacetic acid and 0.2 mg l−1 kinetin gave the best callus induction. Shoots were initiated from the calli with reduced 2,4-D and increased kinetin concentration.

Genetic diversity in important members of Cucurbitaceae using isozyme, RAPD and ISSR markers.

Biochemical and molecular markers have been used on eleven species of Cucurbitaceae collected from lower Gangetic plains. Six enzyme systems were selected. Among 40 primers examined, 14 random amplified polymorphic DNA (RAPD) and 10 inter-simple sequence repeat (ISSR) primers were selected for the analysis. Generated RAPD (100) and ISSR (100) fragments showed high variations among the species. Jaccard similarity coefficients were used for the evaluation of pairwise genetic divergence; cluster analysis of the similarity matrices was performed to estimate interspecific diversity. Further, principal coordinate analysis was performed to evaluate the resolving power of the three marker systems to differenciate among the species.

Genetic Diversity of Amaranthus Species from the Indo-Gangetic Plains Revealed by RAPD Analysis Leading to the Development of Ecotype-Specific SCAR Marker

Genetic diversity and relationships among 6 Amaranthus species from 8 phytogeographic regions of the Indo-Gangetic plains were analyzed using a random amplified polymorphic DNA (RAPD) marker. RAPD primers yielded a total of 262 amplicons, ranging from approximately 250 to approximately 3000 bp in size with an average of 13.1 amplicons per primer, of which 254 amplicons (96.94%) were polymorphic. The genetic similarity coefficient among all the Amaranthus species ranged from 0.16 to 0.97 with a mean similarity coefficient of 0.56, indicating that variation existed in the genetic diversity of different populations. In the unweighted pair group method with arithmetic average dendrogram, populations of the same species clustered together. A unique 1371-bp RAPD band specific for Amaranthus gangeticus (syn. tricolor) of a particular phytogeographic region was converted to a sequenced characterized amplified region (SCAR) marker. The translated marker sequence showed homology with hemagglutinin protein. This SCAR marker is potentially useful for germplasm conservation and identification of amaranth ecotype.

IN VITRO PLANT REGENERATION IN HOLARRHENA ANTIDYSENTERICA WALL., THROUGH HIGH- FREQUENCY AXILLARY SHOOT PROLIFERATION

SummaryAn efficient, rapid and large-scale propagation of the woody, aromatic and medicinal shrub, Holarrhena antidysenterica, through in vitro culture of nodal segments with axillary buds, is described. N6-benzyladenine used at 15 μM was the most effective in inducing bud break and growth, and also in initiating multiple shoot proliferation at the rate of 43 microshoots per nodal explant with axillary buds, after 30 d of eulture. By repeated subculturing of nodal explants with axillary buds, a high-frequency multiplication rate was established. Efficient rooting was achieved with 35 μM indole-3-butyric acid which was the most effective in inducing roots, as 80% of the microshoots produced roots. Plantlets went through a bardening phase in a controlled plant growth chamber, prior to ex vitro transfer Micropropagated plants established in garden soil were uniform and identical to donor plants with respect to growth characteristics and vegetative morphology.

Effects of herbicide 2,4-dinitrophenol on mitosis, DNA, RNA and protein synthesis inNigella sativa L.

Effect of 2,4-dinitrophenol (DNP) was studied onNigella sativa to note the changes in mitosis, DNA, RNA and protein synthesis. The chemical affected division frequency considerably and chromosomal abnormalities like sticky bridge, fragmentation, micronucleietc. were recorded. By using precursors of nucleic acid and protein synthesis, it was found that DNP also inhibited DNA, RNA and protein synthesis. The decrease in division frequency can be correlated with the DNA synthesis.

EFFICIENT PLANT REGENERATION IN HOLARRHENA ANTIDYSENTERICA WALL., FROM SHOOT SEGMENT-DERIVED CALLUS

SummaryA procedure has been outlined for plant regeneration of an important medicinal shrub, Holarrhena antidysenterica, through shoot segment-derived callus. Explants used for callus induction were shoot segments derived from 14-d-old axenic plants on Murashige and Skoog (MS) medium supplemented with 15 μM N6-benzyladenine (BA). A white friable type of callus was obtained in 4.52 μM 2,4-dichlorophenoxyacetic acid and 2.32 μM kinetin which did not have the potentiality to regenerate. High-frequency shoot differentiation was achieved on transferring the friable callus to MS medium supplemented with 17.8 μM BA and 8.0 μM naphthaleneacetic acid. The highest percentage of calluses forming shoots (65.06±2.26) was achieved in this medium. The organogenetic potential of the regenerating callus was influenced by the age of the culture. Rooting was achieved on the shoots using MS medium with 25 μM indolebutyric acid. The plantlets were acclimatized and established in soil. The regenerated plants were morphologically uniform and exhibited similar growth characteristics and vegetative morphology to the donor plants.

Chromosomal variations in the callus tissues ofAllium tuberosum andA. cepa

SummaryChromosome studies ofAllium tuberosum andA. cepa were made from one month to eighteen months old calluses. Different types of chromosomal variations like aneuploid number ranging from 28 to 31, tripolarity, lagging, micronuclei, haploid number etc. were noted inA. tuberosum, whereas inA. cepa the cells showed high chromosome numbers such as 32, 64 or more. The normal chromosome number (2n=16) occurred rarely. The selective pressure of the culture media may have led to the manifestation of the genetic control of differential response to chromosome behaviour and growth in the two species of the same genus.

Effect of different hormones on Vigna tissue culture and its chromosomal behaviour

Abstract Vigna sinensis cv. 152 hypocotyl segments were grown in vitro in the presence of different auxins and cytokinins. The growth pattern was observed at different combinations of hormone. Indoleacetic acid (IAA) was most potent in root initiation but least so in callus induction. Both naphthyl acetic acid (NAA) and 2,4-dichlorophenoxy acetic acid (2,4-D) were almost equally potent in callus initiation but NAA produce healthier calli and caused some root initiation. 2,4-D could not cause any root initiation. Cytological analysis of calli revealed that 2,4-D induces more numerical chromosomal anomalies in Vigna sinensisthan NAA.

In vitro culture of Cuminum cyminum regeneration of flowering shoots from calli of hypocotyl and leaf explants

Calli from hypocotyl explant of Cuminum cyminum L. (Cumin) grew rapidly on Gamborgs B5 basal medium with the following supplements, (i) 0.5 mg/l — 2,4-D (ii) 4 mg/l — NAA plus 2 mg/l — Kinetin and (iii) 0.2 mg/l — NAA plus 0.2 mg/l — BAP, whereas calli from leaf explant in these media grew slowly. Hypocotyl and leaf calli produced roots when transferred to basal medium only and shoots in basal medium with 0.5 mg/l NAA and 0.1 mg/l BAP. Ninety percent of the shoots produced roots when they were transferred to half strength MS inorganic salts supplemented with 0.5 mg/l each of IBA and NAA.Fifty to sixty percent of rootless as well as rooted shoots produced terminal umbellate flowers on this medium.