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Background Embelia ribes Burm f. (Primulaceae) is a medicinal and vulnerable woody liana distributed throughout India. Embelin, a well-recognized active phytoconstituents in berries, is commonly used in ayurvedic formulations. Due to over-exploitation, the status of the plant is vulnerable. Previous studies on this species mainly focused on its phytochemical analysis, which led to overexploitation and loss of the germplasm. Methods and results In the present study, 20 RAPD and 18 ISSR markers were employed to assess genetic divergence in 40 genotypes of E. ribes collected from different parts of the Western Ghats of India. In RAPD analysis, all 40 accessions with 20 RAPD primers amplified 282 fragments, with 83.91% average polymorphism and with an average of 14.10 bands per primer. The size of amplicons varied from 200 to 2500 bp. While, ISSR primers produced 203 fragments of which 161 were polymorphic with an average of 11.28 bands per primer with 73.25% average polymorphism. The size of amplicons ranges from 200 to 2500 bp. RAPD and ISSR markers were also assessed by calculating polymorphic information content (PIC) to discriminate the genotypes; the average PIC value for RAPD, ISSR, and combined RAPD + ISSR markers obtained was more than 0.50 suggesting the informativeness of markers. UPGMA analysis based on Jaccard’s similarity coefficient for RAPD, ISSR, and RAPD + ISSR data reveals that 40 accessions of E. ribes were depicted in four clusters. The clustering pattern of all individuals in PCoA analysis agreed with the UPGMA dendrograms, which further confirms the genetic relationships explained by cluster analysis. AMOVA analysis of RAPD, ISSR, and combined marker system revealed variation within the population, ranging from 41 to 44%, and among the population, it ranged from 56 to 59%. Conclusion The present study provides an optimized method for evaluating the genetic diversity of Embelia ribes using RAPD and ISSR markers which are useful for further sustainable utilization and conservation of natural populations in the Western Ghats of India.

Tea [ Camellia sinensis (L.) Kuntze] has primarily been improved by selections and controlled hybridizations. In India, the genetic improvement programs are largely led by United Planters Association of South India (UPASI). Tea has robust vegetative propagation and several high yielding commercial elite tea clones released by UPASI have been cultivated across the world. In a previous study, we analysed 42 elite UPASI tea clones using cytological and molecular analysis (Sharma and Raina, Int J Tea Sci 5:21–28, 2006 ). Present work analysed the same clones using Random amplified polymorphic DNA (RAPD) and Inter simple sequence repeat (ISSR) markers to document the genetic diversity and delineate the genetically distinct superior tea clones. A total of 447 and 116 bands were generated with 52 RAPD and 27 ISSR primers, out of which 395 and 70 bands, respectively were observed to be polymorphic. RAPD markers outcompeted ISSRs when compared against various genetic diversity attributes. An overall low Nei’s gene diversity (0.23 and 0.19) and higher value of gene flow (6.5 and 5.0) with both markers indicated narrow genetic base for the clones. Dendrograms delineated 42 clones into three major clusters whereas population STRUCTURE analysis clustered them into 6 subpopulations without discrete morphotype based grouping. Presence of many admixtures in STRUCTURE indicates towards diverse genetic ancestry of the analysed tea clones. A high level of genetic variation (90.48%) was revealed with analysis of molecular variance (AMOVA) within populations as compared to a low (9.52%) level among populations. A few superior clones were found to be genetically distinct than others and can be fruitfully used in future tea breeding programme.

Gloriosa superba L., an endangered medicinal plant with global interest due to presence of colchicine, an important alkaloid used in formulations of Indian and Traditional medicine. The plant has become endangered due to its unscientifically exploitation and high medicinal values. In the Present study 10 randomly amplified polymorphic DNA (RAPD) and 6 ISSR markers were employed to assess genetic divergence among micro propagated, wild and field cultivated plants of Gloriosa superba collected from different parts of India. In RAPD analysis, all the 10 accession with 10 RAPD primers amplified 466 fragments, with 96.43 % polymorphism and with an average of 46.6 bands per primer. The size of amplicons varied from 1656 to 100 bp. While, ISSR primers produced 328 fragments of which 298 were polymorphic with an average of 49.7 bands per primer with 91.83% polymorphism. The size of amplicons ranges from 2395 to 181 bp. RAPD, ISSR markers were also assessed by calculating polymorphic information content (PIC) to discriminate the genotypes, Average PIC value for RAPD, ISSR and combined RAPD + ISSR markers obtained was ≤ 0.50 suggesting the informativeness of markers. Jaccard’s coefficient ranges from 0.18 to 0.75 (RAPD) and 0.17 to 0.61 (ISSR) and 0.21–0.52 for pooled ISSR and RAPD markers. The clustering pattern based on UPGMA analysis of the genotypes in the combined analysis revealed that the majority of the genotypes remained similar to the ISSR dendrogram, while the RAPD-based dendrogram showed some variation in the clustering of genotypes. The result of PCA scattered plot obtained were in agreement with the UPGMA dendrogram, which further confirms the genetic relationships explain by cluster analysis. Results confirmed that the genotype studied had good genetic diversity and can be used for identification, conservation, and future breeding program of Gloriosa species and consequently for the benefit of the pharmaceutical industries.

Castor (Ricinus communis L.), known as castor oil plant or castor bean, is a non-edible oilseed crop. In the present study, the genetic diversity among 54 samples (3 wild and 51 cultivated) collected worldwide was evaluated using inter-simple sequence repeats (ISSRs) and random amplified polymorphic DNA (RAPD) markers. A total of 9 ISSR primers produced 83 high-resolution bands with 61 (74.53%) as polymorphic. The percentage of polymorphic bands per primer and the genetic similarity coefficient ranged from 54.55% (UBC-836) to 100% (UBC-808) and from 0.74 to 0.96, respectively. A total of 11 out of 20 RAPD primers amplified unique polymorphic products with an average percentage of polymorphic bands of 60.98% (56 polymorphic bands out of a total of 90 bands obtained). The percentage of polymorphic bands per primer ranged from 25% (OPA-02 and B7) to 90.91% (B21) with the genetic similarity coefficient ranging from 0.73 to 0.98. The unweighted pair group method with arithmetic averages (UPGMA) dendrogram using two molecular markers divided 54 castor genotypes into three groups. Furthermore, based on morphological data, all 54 castor varieties were grouped into three main clusters. The genetic diversity analysis based on two molecular makers showed that most varieties from China were closely related to each other with three varieties (GUANGDONGwild, ZHEJIANGWild, and HANNANWild) belonging to a wild group separated from most of the cultivated castor samples from China, India, France, and Jordan. These results suggested that the cultivated castor contains a narrow genetic base. Accordingly, we recommend that wild castor genetic resources be introduced for breeding novel castor varieties. Furthermore, the Vietnam, Malaysia, Indonesia, and Nigeria accessions were clustered into the same group. The results of principal coordinate analysis (PCoA) and UPGMA cluster analysis were consistent with each other. The findings of this study are important for future breeding studies of castor.

The use of molecular genetic techniques is becoming increasingly widespread in analyses of hybrid zones. Yet, exactly how many molecular markers are required to provide a given degree of resolution remains an open question. We present statistical models that relate the number of markers examined to their power to discriminate between pure species, F1's, and backcross individuals. Our models indicate that only about four or five markers are required to provide a coarse classification of individuals in hybrid zones, whereas upwards of 70 markers are required to discriminate between pure species and advanced backcrosses. The models provide hybrid zone researchers a basis upon which to balance the time and expense of examining large numbers of markers against the magnitude of classification errors when smaller numbers of markers are examined.

In traditional medicine, Lonicera japonica (Thunb.) has a notable place, and it has been used for thousands of years in China, Japan, Korea and other East-Asian countries for treating cancer, inflammation, hepatic complications, influenza and wounds. However, the molecular or genetic characteristic of this plant is not well defined. In this study, improved random amplified polymorphic DNA (RAPD) has been employed for the genetic characterization of five varieties of L. japonica collected from different geographic locations of Southern China. A total of 147 bands of DNA fragments were obtained in RAPD-PCR by using 18 primers, and the band sizes ranged from approximately 300–2,000 bp, with 3–11 amplified bands for each primer. Based on the RAPD amplification profiles, cluster dendrogram was obtained, which showed that the similarity coefficients among five varieties of L. japonica ranged from 0.59 to 0.77. To our knowledge, this is the first report of genetic characterization of L. japonica using improved RAPD analysis which has been validated by ISSR analysis, and this characterization may be useful for the preservation of genetic diversity and Lonicera population identification. Moreover, as an option, the improved method could be employed for a variety of applications in genetic diversity and fingerprinting analyses.

A total of 10 endophytic actinomycete strains were successfully isolated from healthy shoots and roots of Aquilaria crassna Pierre ex Lec (eaglewood). Analysis of 16S rDNA sequencing of those isolates showed that they belong to members of the genera Streptomyces (2 isolates), Nonomuraea (1 isolate), Actinomadura (1 isolate), Pseudonocardia (1 isolate) and Nocardia (3 isolates). The remaining 2 isolates were unidentified. All of isolates produced the amount of indole-3-acetic acid (IAA) and ammonia ranging between 9.85 ± 0.31 to 15.14 ± 0.22 μg ml −1 and 2 to 60 mg ml −1 , respectively. Among 10 isolates tested, the amount of hydroxamate-type siderophore produced by 2 isolates was undetectable. While the remaining 8 isolates produced the amount of hydroxamate-type ranging between 3.21 ± 0.12 and 39.30 ± 0.40 μg ml −1 . Also, catechols-type siderophore produced by 9 isolates was undetectable. Actinomadura glauciflava is only one isolate that produced catechols-type 4.12 ± 0.90 μg ml −1 . In addition, 10 endophytic actinomycetes showed protease activity ranging from undetectable to 8.16 ± 0.15 unit ml −1 . Genetic relatedness amongst these isolates was determined base on Random amplified polymorphic DNA (RAPD) and Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC PCR). Both methodologies generated specific patterns corresponding to particular genotypes. RAPD fingerprinting proved to be slightly more discriminatory than ERIC PCR. This study is the first published report that actinomycetes can be isolated as endophytes within this plant. It is also the first published report that endophytic actinomycetes are capable of producing IAA and siderophores.

The aim of this study was to evaluate the occurrence of yeast populations during different olive oil extraction processes, carried out in three consecutive years in Tuscany (Italy), by analysing crushed pastes, kneaded pastes, oil from decanter and pomaces. The results showed yeast concentrations ranging between 10 3 and 10 5  CFU/g or per mL. Seventeen dominant yeast species were identified by random amplified polymorphic DNA with primer M13 and their identification was confirmed by restriction fragments length polymorphism of ribosomal internal transcribed spacer and sequencing rRNA genes. The isolation frequencies of each species in the collected samples pointed out that the occurrence of the various yeast species in olive oil extraction process was dependent not only on the yeasts contaminating the olives but also on the yeasts colonizing the plant for oil extraction. In fact, eleven dominant yeast species were detected from the washed olives, but only three of them were also found in oil samples at significant isolation frequency. On the contrary, the most abundant species in oil samples, Yamadazyma terventina , did not occur in washed olive samples. These findings suggest a phenomenon of contamination of the plant for oil extraction that selects some yeast species that could affect the quality of olive oil.

Jatropha curcas L., a multipurpose shrub has acquired significant economic importance for its seed oil which can be converted to biodiesel, is emerging as an alternative to petro-diesel. The deoiled seed cake remains after oil extraction is toxic and cannot be used as a feed despite having best nutritional contents. No quantitative and qualitative differences were observed between toxic and non-toxic varieties of J. curcas except for phorbol esters content. Development of molecular marker will enable to differentiate non-toxic from toxic variety in a mixed population and also help in improvement of the species through marker assisted breeding programs. The present investigation was undertaken to characterize the toxic and non-toxic varieties at molecular level and to develop PCR based molecular markers for distinguishing non-toxic from toxic or vice versa. The polymorphic markers were successfully identified specific to non-toxic and toxic variety using RAPD and AFLP techniques. Totally 371 RAPD, 1,442 AFLP markers were analyzed and 56 (15.09%) RAPD, 238 (16.49%) AFLP markers were found specific to either of the varieties. Genetic similarity between non-toxic and toxic verity was found to be 0.92 by RAPD and 0.90 by AFLP fingerprinting. In the present study out of 12 microsatellite markers analyzed, seven markers were found polymorphic. Among these seven, jcms21 showed homozygous allele in the toxic variety. The study demonstrated that both RAPD and AFLP techniques were equally competitive in identifying polymorphic markers and differentiating both the varieties of J. curcas. Polymorphism of SSR markers prevailed between the varieties of J. curcas. These RAPD and AFLP identified markers will help in selective cultivation of specific variety and along with SSRs these markers can be exploited for further improvement of the species through breeding and Marker Assisted Selection (MAS).

The diverse metabolites, positively affecting the nutritional, organoleptic and technological traits of leavened baked goods, are produced by different sourdough lactic acid bacteria (LAB) and yeast strains, as the result of their genetic intraspecific diversity. Therefore, the molecular and functional strain-level characterization of sourdough microbiota is crucial to valorize traditional or origin protected baked end-products, develop innovative starter cultures and design functional cereal-based foods. To this aim, the genetic intraspecific diversity of 96 Lactobacillus sanfranciscensis, 65 Kazachstania humilis and three Saccharomyces cerevisiae characterizing Protected Designation of Origin (PDO) Tuscan bread sourdough, was investigated, using P4, P7 and M13 random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), (GTG)5 repetitive element sequence-based (rep)-PCR and inter-delta region analyses, respectively. Regarding LAB, the combination of P4, P7 and M13 RAPD-PCR analyses revealed a huge degree of intraspecific variability, discriminating 43 biotypes out of 96 isolates of L. sanfranciscensis. (GTG)5 rep-PCR showed a discriminatory index of 0.95, grouping the 65 K. humilis isolated from PDO Tuscan bread sourdough in 9 biotypes. The high polymorphism among both LAB and yeast isolates of PDO Tuscan bread sourdough outlines a highly complex microbial community structure, whose relative composition and specific physiological characteristics could be responsible for the peculiar organoleptic, rheological, nutritional and potentially nutraceutical features of PDO Tuscan bread.