National Botanical Research Institute Research Articles

Antimicrobial screening of ethnobotanically important stem bark of medicinal plants

Background:The stem barks are the rich sources of tannins and other phenolic compounds. Tannins inhibited the growth of various fungi, yeast, bacteria and virus. Hence, ten stem barks of ethnomedicinally important plants were screened for antibacterial and antifungal activities against human pathogenic strains.Methods:Air-dried and powdered stem bark of each plant was extracted with 50% aqueous ethanol, lyophilized and the dried crude extracts were used for the screening against 11 bacteria and 8 fungi. Antibacterial and antifungal activities were performed according to microdilution methods by NCCLS.Results:The plants Prosopis chilensis, Pithecellobium dulce, Mangifera indica showed significant antibacterial and antifungal activities against Streptococcus pneumonia, Enterobacter aerogenes, Klebsiella pneumonia and Candida albicans with MIC of 0.08mg/ml. Pithecellobium dulce bark also showed significant antibacterial activity against Bacillus cereus.Conclusion:The bark of Pithecellobium dulce has more or less similar activity against the known antibiotic and may be considered as potent antimicrobial agent for various infectious diseases.

Phosphate solubilization potential and phosphatase activity of rhizospheric trichoderma spp.

Trichoderma sp., a well known biological control agent against several phytopathogens, was tested for its phosphate (P) solubilizing potential. Fourteen strains of Trichoderma sp. were isolated from the forest tree rhizospheres of pinus, deodar, bamboo, guava and oak on Trichoderma selective medium. The isolates were tested for their in-vitro P-solubilizing potential using National Botanical Research Institute Phosphate (NBRIP) broth containing tricalcium phosphate (TCP) as the sole P source, and compared with a standard culture of T. harzianum. All the cultures were found to solubilize TCP but with varying potential. The isolate DRT-1 showed maximum amount of soluble phosphate (404.07 εg.ml-1), followed by the standard culture of T. harzianum (386.42 εg.ml-1) after 96 h of incubation at 30±10C. Extra-cellular acid and alkaline phosphatases of the fungus were induced only in the presence of insoluble phosphorus source (TCP). High extra-cellular alkaline phosphatase activity was recorded for the isolate DRT-1 (14.50 U.ml-1) followed by the standard culture (13.41 U.ml-1) at 72h. The cultures showed much lesser acid phosphatase activities. Under glasshouse conditions, Trichoderma sp. inoculation increased chickpea (Cicer arietinum) growth parameters including shoot length, root length, fresh and dry weight of shoot as well as roots, in P-deficient soil containing only bound phosphate (TCP). Shoot weight was increased by 23% and 33% by inoculation with the isolate DRT-1 in the soil amended with 100 and 200 mg TCP kg-1 soil, respectively, after 60 d of sowing. The study explores high P-solubilizing potential of Trichoderma sp., which can be exploited for the solubilization of fixed phosphates present in the soil, thereby enhancing soil fertility and plant growth.

Characterization of tricalcium phosphate solubilization by Stenotrophomonas maltophilia YC isolated from phosphate mines

The phosphate solubilizing characteristics of a strain YC, which was isolated from phosphate mines (Hubei, China), were studied in National Botanical Research Institute’s phosphate (NBRIP) growth medium containing tricalcium phosphate (TCP) as sole phosphorus (P) source. The strain YC is identified as Stenotrophomonas maltophilia (S. maltophilia) based upon the results of morphologic, physiological and biochemical characteristics and 16S rRNA sequences analysis. The results show that the strain S. maltophilia YC can solubilize TCP and release soluble P in NBRIP growth medium. A positive correlation between concentration of soluble P and population of the isolate and a negative correlation between concentration of soluble P and pH in the culture medium are observed from statistical analysis results. Moreover, gluconic acid is detected in the culture medium by HPLC analysis. It indicates that the isolate can release gluconic acid during the solubilizing experiment, which causes acidification of the culture medium and then TCP solubilization. S. maltophilia YC has a maximal TCP solubilizing capability when using maltose as carbon source and ammonium nitrate as nitrogen source, respectively, in NBRIP growth medium.

Comparison of media used to evaluateRhizobium leguminosarumbivarviciaefor phosphate-solubilizing ability

Rhizobium leguminosarum is well known for its ability to fix nitrogen (N). In addition, its capacity to solubilize phosphate (Ph) has been receiving attention in recent years. Our ultimate objective was to select a R. leguminosarum bv. viciae isolate with superior Ph-solubilizing ability. The first step was to identify a culture medium that is sensitive and effective in identifying the ability of R. leguminosarum bv. viciae isolates to solubilize Ph. Thirty isolates were evaluated for Ph solubilization in broth and on solid formulations of three media: yeast mannitol extract (YEM), National Botanical Research Institute phosphate nutrient medium (MNBRI), and Pikovskaya phosphate medium (PVK). All media contained 5 g/L CaHPO4 as the only phosphorus (P) source. All 30 isolates increased the Ph concentration in liquid cultures, but the amount of Ph released into solution by individual isolates varied from one medium to another. In contrast, only a subset of the 30 isolates solubilized Ph on the solid cultures. Furthermore, some of the isolates that were able to solubilize Ph were only able to do so on a single medium. Regression analysis revealed no relationship between the Ph concentration in the liquid media and the zones of clearing on the solid media (p > 0.05). Although the pH of all of the liquid media dropped after 12 days of growth of the isolates, a relationship between Ph concentration and pH existed only for the MNBRI medium (r2 = 0.485, p < 0.001). Increasing the amount of N in the MNBRI medium from 0.1 g/L of (NH4)2SO4 to 0.5 g/L of (NH4)2SO4 did not affect the amount of Ph in solution, but it profoundly reduced the survival of the R. leguminosarum by approximately 50-fold. Consequently, the surviving bacteria were either more efficient at solubilizing Ph in the high N media or organic acids released from the lysis of the dead cells solubilized the CaHPO4 source.

Isolation of Phosphate-Solubilizing Fungi from Phosphate Mines and Their Effect on Wheat Seedling Growth

Three phosphate-solubilizing fungi, identified as Penicillium expansum, Mucor ramosissimus, and Candida krissii, were isolated from phosphate mines (Hubei, People's Republic of China) and characterized. All the isolates demonstrated diverse levels of phosphate-solubilizing capability in National Botanical Research Institute's phosphate growth medium containing rock phosphate as sole phosphate source. Acidification of culture medium seemed to be the main mechanism for rock phosphate solubilization. Indeed, citric acid, oxalic acid, and gluconic acid were shown to be present in the culture medium inoculated with these isolates. Moreover, the isolates produced acid and alkaline phosphatases in culture medium, which may also be helpful for RP solubilization. A strong negative correlation between content of soluble phosphorus and pH (r = - 0.89; p < 0.01) in culture medium was observed in this study. All the isolates promoted growth, soil available phosphorus, phosphorus, and nitrogen uptake of wheat seedling in field soil containing rock phosphate under pot culture conditions, thus demonstrating the capability of these isolates to convert insoluble form of phosphorus into plant available form from rock phosphate, and therefore hold great potential for development as biofertilizers to enhance soil fertility and promote plant growth.

Isolation and characterisation of phosphate solubilising microorganisms from the cold desert habitat of Salix alba Linn. in trans Himalayan region of Himachal Pradesh

Phosphate solubilising microorganisms (PSM) (bacteria and fungi) associated with Salix alba Linn. from Lahaul and Spiti valleys of Himachal Pradesh were isolated on Pikovskaya (PVK), modified Pikovskaya (MPVK) and National Botanical Research Institute agar (NBRIP) media by spread plating. The viable colony count of P-solubilising bacteria (PSB) and fungi (PSF) was higher in rhizosphere than that of non-rhizosphere. The frequency of PSM was highest on MPVK followed by NBRIP and PVK agar. The maximum proportion of PSM out of total bacterial and fungal count was found in upper Keylong while the least in Rong Tong. The PSB frequently were Gram-positive, endosporeforming, motile rods and belonged to Bacillus sp. The PSF mainly belonged to Penicillium sp., Aspergillus fumigatus, A. niger, A. spp. and non-sporulating sterile. Amongst the isolates with high efficiency for tricalcium phosphate (TCP) solubilisation, seven bacterial and seven fungal isolates dissolved higher amount of P from North Carolina rock phosphate (NCRP) than Mussoorie rock phosphate (MRP) and Udaipur rock phosphate (URP). However, the organisms solubilised higher-P in NBRIP broth than PVK broth. SBC5 (Bacillus sp.) and SBC7 (Bacillus sp.) bacterial isolates exhibited maximun P solubilisation (40 and 33 μg ml(-1) respectively) whereas FC28 (Penicillium sp.) isolate (52.3 μg ml(-1)) amongst fungi while solubilising URP. The amount of P solubilised was positively correlated with the decrease in pH of medium. SBC5 (Bacillus sp.), SBC7 (Bacillus sp.) and SBC4 (Micrococcus) decreased the pH of medium from 6.8 to 6.08 while FC28 (Penicillium sp.) and FC39 (Penicillium sp.) isolates of fungi recorded maximum decrease in pH of medium from 6.8 to 5.96 in NBRIP broth.

Optimization for rock phosphate solubilization by phosphate-solubilizing fungi isolated from phosphate mines

The optimization for rock phosphate (RP) solubilization by phosphate-solubilizing fungi ( Candida krissii, Penicillium expansum and Mucor ramosissimus) isolated from phosphate mines (Hubei, PR China) was investigated. The content of soluble phosphorus (P) released by these isolates was tested on the National Botanical Research Institute's phosphate growth medium (NBRIP) containing RP as sole P source. Results showed that the optimum conditions were at temperature, 32 °C; shaking speed, 160 rpm; RP concentration, 2.5 g l −1; EDTA concentration, 0.5 mg ml −1. The content of soluble P gradually decreased with a larger particle size of RP with all the isolates. The content of soluble P was the highest when the initial pH for RP solubilization was 5.5 in the medium inoculated with C. krissii, which was different from that of 7.0 in the medium inoculated with P. expansum and 7.5 in the medium inoculated with M. ramosissimus. All the isolates showed lower tolerance with increasing concentration of silver ion (Ag +). Moreover, the highest content of soluble P was achieved when released by C. krissii at 20 of C:N ratio, P. expansum at 40 of C:N ratio and M. ramosissimus at 30 of C:N ratio.

Identification of a potyvirus associated with mosaic disease of Narcissus sp. in India

Narcissus tazetta cv. Paperwhite is an important ornamental for the floriculture industry. Narcissus is susceptible to three members of genus Potyvirus, namely Narcissus yellow stripe virus, Narcissus late season yellows virus and Narcissus degeneration virus, plus Narcissus latent virus (genus Macluravirus, family Potyviridae). A disease characterized by severe mosaic and light green to yellow leaf stripes, was observed on Narcissus in an experimental plot at the National Botanical Research Institute, Lucknow. Affected plants also showed extreme reduction in their growth, and loss of yield and bloom quality. Total RNA was extracted from 100 mg of leaf tissue as described by Gibbs & MacKenzie (1997). RT-PCR was performed with potyvirid specific primers Pot1 and Pot2 (Gibbs & MacKenzie, 1997) to amplify part of the NIb gene, the coat protein (CP) gene and the 3′ untranslated region (UTR). The amplified DNA fragment of c. 1·5 kb was cloned into the p-drive cloning vector (QIAGEN) and sequenced (GenBank accession DQ991145). The CP gene nucleotide sequence showed 98% identity with Chinese narcissus potyvirus Chongming isolate (AJ311374·1), 97% with Chinese narcissus potyvirus Zhangzhou isolate (AJ311373·1) and 95% with Lycorsis potyvirus (AF511486·1). The 3′-UTR (148 nt.) shared 100% identity with the Chinese narcissus Chongming and Zhangzhou isolates and 97% identity with Lycoris potyvirus (AF511486·1). Earlier investigations in India (Aminuddin et al., 1989, 1999), based mainly on biological, serological and electron microscopic properties demonstrated the association of an unidentified potyvirus with severe mosaic disease of Narcissus tazetta. However, nucleotide sequence information (CP gene and 3′ UTR) available in this study clearly established the identity of the associated potyvirus, at the species level, as the virus first recorded as Lycoris potyvirus. This is the first specific identification of a potyvirus from narcissus in India. However, Narcissus spp. are prone to mixed viral infections (Chen et al., 2003) so this potyvirus may not be the sole agent causing the mosaic disease.

Varietal identification and assessment of genetic relationships in Hippeastrum using RAPD markers

The National Botanical Research Institute (NBRI) in Lucknow, India, maintains germplasm of Hippeastrum, a beautiful summer blooming ornamental. Germplasm collections comprise NBRI hybrids developed through selective breeding, hybrids with unknown parentage, local species, and Dutch hybrids for research purposes. Considering the importance of protecting plant breeders’ rights for commercial exploitation of hybrids, a PCR-based technique (random amplified polymorphic DNA—RAPD) was used to correctly identify known and unknown hybrids and to determine cultivar relatedness. RAPD profiles were used very successfully to trace and confirm the parentage of all the hybrids tested and to determine clear molecular relationships among varieties.

Phytoremediation of a sodic forest ecosystem: plant community response to restoration process.

The creation of a new biotope on degraded lands is the focused objective nowadays, on a global level. An attempt was made to rehabilitate sodic waste lands through the establishment of plant cover, with diverse plant communities within Banthra Research Station of the National Botanical Research Institute, Lucknow, India (80degr45min-53degr53min E and 26degr40min- 26degr45min N) for the last three decades. A rehabilitated forest ecosystem developed in this way consists in a number of herbs, shrubs and trees. Derris indica, Dalbergia sissoo, Azadirachta indica, Cassia siamea, Terminalia arjuna, Syzigium cumin were the dominant species in this rehabilitated forest, whereas Sporobolus, Desmostachya and Dactyloctenium were a common genera of grasses /forbs on a barren land. This study showed that species diversity and productivity significantly influenced the soil amelioration process. A comparison of three afforested sites with a varying degree of productivity and diversity indicated that some soil properties were influenced by diversity while others by productivity. Individual effects of diversity and productivity were classified as 42% and respectively 58%, in soil reclamation. A combined effect of both biomass productivity and species diversity contributed about 92% towards amelioration. There was an appreciable reduction in soil pH and ESP (exchangeable sodium percentage) and an increase in organic C and N contents showing that sodicity has receded in the forested soil. This experience can be tried out on similar sites of arid and semiarid regions of the world for the bioreclamation of sodic lands.

Isolation and Identification of Potential Phosphate Solubilizing Bacteria from the Rhizoplane of Oryza sativa L. cv. BR29 of Bangladesh

A total of 30 bacteria were isolated from the rhizoplane of rice cv. BR29 cultivated in Mymensingh, Bangladesh and from the seedlings obtained from surface-sterilized seeds of BR29. Upon screening, 6 isolates showed varying levels of phosphate solubilizing activity in both agar plate and broth assays using National Botanical Research Institute's phosphate medium. The bacterial isolates were identified based on their phenotypic and 16S rRNA genes sequencing data as Acinetobacter sp. BR-12, Klebsiella sp. BR-15, Acinetobacter sp. BR-25, Enterobacter sp. BR-26, Microbacterium sp. BRS-1 and Pseudomonas sp. BRS-2. The BR-25 exhibited highest phosphate solubilizing activity followed by BR-15. They grew rapidly in the liquid medium at pH 5 and 7 but almost no growth occurred at pH 3. The pH value of the culture medium was decreased with bacterial growth suggesting that they might secrete organic acids to solubilize insoluble phosphorus. Scanning electron microscope analysis of two-week-old rice seedlings germinated from seeds previously inoculated with BR-25 and BR-15 revealed dense colonization at the root surfaces presumably using fimbriae on the bacterial cells.

Ethyl Methanesulfonate Mutagenesis–Enhanced Mineral Phosphate Solubilization by Groundnut-Associated Serratia marcescens GPS-5

Twenty-three bacterial isolates were screened for their mineral phosphate-solubilizing (MPS) ability on Pikovskaya and National Botanical Research Institute's phosphate (NBRIP) agar. The majority of the isolates exhibited a strong ability to solubilize hydroxyapatite in both solid and liquid media. The solubilization in liquid medium corresponded with a decrease in the pH of the medium. Serratia marcescens GPS-5, known for its biocontrol of late leaf spot in groundnut, emerged as the best solubilizer. S. marcescens GPS-5 was subjected to ethyl methanesulfonate (EMS) mutagenesis, and a total of 1700 mutants, resulting after 45 minutes of exposure, were screened on buffered NBRIP medium for alterations in MPS ability compared with that of the wild type. Seven mutants with increased (increased-MPS mutants) and 6 mutants with decreased (decreased-MPS mutants) MPS ability were isolated. All seven increased-MPS mutants were efficient at solubilizing phosphate in both solid and liquid NBRIP medium. Among the increased-MPS mutants, EMS XVIII Sm-35 showed the maximum (40%) increase in the amount of phosphate released in liquid medium compared with wild-type S. marcescens GPS-5, therefore, it would be a useful microbial inoculant in groundnut cultivation. EMS III Sm W, a nonpigmented mutant, showed the lowest solubilization of phosphate among the 6 decreased-MPS mutants.

A Yellow Mosaic Disease of Soybean in Northern India is Caused by Cotton leaf curl Kokhran virus.

Soybean, Glycine max (L.) Merr., is a protein- and oil-rich crop cultivated in India and abroad. A yellow mosaic disease was observed on soybean with 80 to 90% disease incidence during August 2005 at fields of the National Botanical Research Institute, Lucknow, in northern India. Soybean plants were found to be infested with whiteflies (Bemisia tabaci) suggesting begomovirus etiology. The disease agent was transmitted experimentally by whiteflies, and symptoms developed after 23 days. Total DNA was isolated from 51 leaf samples collected from 42 symptomatic and 9 asymptomatic plants. Polymerase chain reaction was performed using begomovirus coat protein-specific primers 5'-ATGGCGAA GCGACCAG-3' and 5'-TTAATTTGTGACCGAATCAT-3' (AM180920/ AM180921). An amplicon of the expected size (~800 bp) was obtained in all 42 symptomatic leaves but not from any of the nine asymptomatic leaf samples. The amplicon was cloned, and the identical sequence of three clones was submitted to GenBank (Accession No. DQ343283). BLAST search of nucleotide sequences revealed 95% identity with Cotton leaf curl Kokhran virus (CLCKV) (GenBank Accession Nos. AJ002449, AJ002448, AJ496286, and AY456683) and 57% identity with Mungbean yellow mosaic India virus (MYMIV-Sb, GenBank Accession No. AY049772). Results indicated that the virus associated with yellow mosaic disease of soybean is an isolate of CLCKV rather than MYMIV-Sb (1) reported earlier on soybean from northern India. To our knowledge, this is the first report of soybean as a new host of Cotton leaf curl Kokhran virus. Reference: (1) K. S. Usharani et al. Curr. Sci. 86:845, 2004.

Multivariate analysis and clustering of Cuphea procumbens inbred lines

15 C. procumbens inbred lines of different eco-geographical origin maintained at National Botanical Research Institute, Lucknow, India were evaluated for seed yield and its 6 component traits to assess genetic divergence among them. The standardized mean values of different traits were subjected to principal component analysis and cluster analysis was performed based on two different clustering strategies i.e. UPGMA and Wards. Ward's method that showed relatively high cophenetic correlation coefficient and significant Wilk's Lambda was identified as the best clustering solution. The first four principal components (PC) with eigenvalues &gt;1 contributed 91.56% of variability among the inbred. First PC was related with fruits/plant and branches/plant; second PC with yield/ plant, seeds/fruit and test weight third PC with plant height; and fourth PC with days to flowering. The genotypes were grouped into five clusters and cluster II was largest with 5 genotypes followed by clusters I, III and Clusters IV, V. Cluster IV exhibited highest mean for seed yield (14.77g) followed by cluster III (14.53g) and the former incorporated inbred lines, NBCP-53 and NBCP-58 that were highly divergent among themselves and from genotypes in other clusters. The inbred in cluster IV and cluster III with good amount of genetic divergence and superior agronomic traits were identified as promising inbred to develop superior recombinants with desirable agronomic traits.

The role of a resource center in national development: The Namibian context

The role of a resource center in national development: The Namibian context

Tree species diversity and dominance in a man-made forest on sodic wasteland of North India

The study was conducted with the objective of studying tree species diversity and dominance and the associated changes in soil characteristics in a man-made forest established on formerly barren sodic land at Banthra Research Station (National Botanical Research Institute, Lucknow, India) (80° 45–53′ E, 26° 40–45′N) over three decades. The results revealed that the forest has a moderate value for the tree species diversity index (H). The tree species Derris indica, Dalbergia sissoo, Azadirachta indica, Cassia siamea, Terminalia arjuna, Syzygium cumini, and Tectona grandis were found to be the major dominant species which may be considered suitable for planting on such degraded wastelands. There was a perceptible reduction in soil pH and exchangeable sodium percentage (ESP) and an increase in organic C and Ca2++Mg2+ cation contents over the past three decades, indicating that the sodicity has declined in the surface soil.

Multivariate analysis in relation to breeding system in opium popy, Papaver somniferum L.

The opium poppy (Papaver somniferum L.) is an important medicinal plant of great pharmacopoel uses. 101 germplasm lines of different eco-geographical origin maintained at National Botanical Research Institute, Lucknow were evaluated to study the genetic divergence for seed yield/plant, opium yield/plant and its 8 component traits following multivariate and canonical analysis. The genotypes were grouped in 13 clusters and confirmed by canonical analysis. Sixty eight percent genotypes (69/101) were genetically close to each other and grouped in 6 clusters (II, III, IV, V, VIII, XII) while apparent diversity was noticed for 32 percent (32/101) of the genotypes who diversed into rest 7 clusters (I, VI, VII, IX, X, XI, XIII). Inter cluster distance ranged from 47.28 to 234.55. The maximum was between IX and X followed by VII and IX (208.30) and IX and XI (205.53). The genotypes in cluster IX, X. XI, and XII had greater potential as breeding stock by virtue of high mean values of one or more component characters and high statistical distance among them. Based on findings of high cluster mean of component trait and inter-cluster distance among clusters, a breeding plan has been discussed.

The natural occurrence of a begomovirus in sunn hemp (Crotalaria juncea) in India

Sunn hemp (Crotalaria juncea) is cultivated in India and other tropical countries as a fibre or green manure crop. During July/August 2001, a severe virus-like disease affected 80–90% of sunn hemp plants in experimental plots at the National Botanical Research Institute, Lucknow. The symptoms of the disease consisted of interveinal chlorosis, yellowing, curling and shortening of leaves, and stunting of plants. Whiteflies (Bemisia tabaci) transmitted the associated virus from diseased to healthy sunn hemp plants which developed symptoms similar to those occurring in naturally infected plants. Total DNA was isolated from the infected-leaf tissues and subjected to polymerase chain reaction (PCR) using begomovirus specific universal primers (AV494/AC1048) which were designed from the conserved sequences of the core region of the coat protein (CP) genes of several begomoviruses (Wyatt & Brown, 1996). As expected, a fragment of c. 500 bp was amplified from the DNA extracted from diseased, but not healthy, leaves. The authenticity of the PCR-amplified DNA fragment was confirmed by Southern hybridization in which DNA probes, prepared from the CP gene of Indian tomato leaf curl virus (IToLCV; Srivastava et al., 1995), cross hybridized with the PCR fragment under high stringency conditions. Although Sunn-hemp mosaic virus and some strains of Tobacco mosaic virus infect sunn hemp (Brunt et al., 1996), a begomovirus has not hitherto been reported to naturally infect this important crop species. Transmission of the sunn hemp virus by whiteflies, PCR amplification of its DNA with begomovirus specific primers and its strong hybridization with a DNA-A probe of IToLCV strongly indicate that it is a begomovirus. The extent of its relationship to IToLCV (Family Geminiviridae; Genus Begomovirus) and other begomoviruses is now being further investigated.

Collar rot of Chlorophytum borivilianum caused by Corticium rolfsii: a new disease

A new disease of Chlorophytum borivilianum, characterized by symptoms of collar rot, has been observed in experimental farms of the National Botanical Research Institute, Lucknow, India. The pathogen has been isolated and identified as Corticium rolfsii.

Molecular distinction amongst varieties of Mulberry using RAPD and DAMD profiles

BackgroundMulberry trees are the most important host for rearing mulberry silkworms in sericulture. Improved varieties of mulberry tree have been developed through traditional breeding procedures. Not much work, however, has been carried out on the molecular characterization of these varieties. Random Amplified Polymorphic DNA (RAPD) and Directed Amplification of Minisatellite DNA (DAMD) methods based on Polymerase Chain Reaction are important tools to analyze genetic diversity of mulberries. These have been used to determine variation amongst nine varieties of Morus spp. maintained at Banthra Research Station of National Botanical Research Institute, Lucknow.Results and DiscussionThe varieties were analyzed using 23 arbitrary sequence decamer primers for RAPD, and 3 minisatellite core sequence primers for DAMD reactions. The RAPD and DAMD band data, (a total of 200 bands), were used to determine the pair wise distances according to Jaccard's algorithm. From these distance values Neighbour Joining (NJ) analyses were carried out separately for the RAPD and the DAMD data. The triploid varieties were found to be most similar to each other using RAPD analysis, while the varieties S13 and S34 were more similar using DAMD analysis. Nearly 85% of the RAPD bands and 91% of the DAMD bands were polymorphic across the nine varieties.ConclusionsThe mulberry varieties could be distinguished by their RAPD and DAMD profiles. As many as five RAPD primers and one DAMD primer generated profiles that can together differentiate all the nine varieties in terms of unique bands.