Numerical Taxonomy And Multivariate Analysis System Research Articles

Genetic differentiation among Batak fish populations (Neolissochilus sumatranus, Tor douronensis, and Tor soro) in North Sumatra, Indonesia revealed by RAPD markers

Abstract. Barus TA, Wahyuningsih H, Simanjuntak BN, Ginting RH, Batubara AS, Hartanto A. 2023. Genetic differentiation among Batak fish populations (Neolissochilus sumatranus, Tor douronensis, and Tor soro) in North Sumatra, Indonesia was revealed by RAPD markers. Biodiversitas 24: 1327-1332. Mahseer or Batak fish species within the genera of Neolissochilus and Tor are highly valued as a source of food for local communities, especially in North Sumatra. Assessment of genetic differentiation of the Batak fish population, namely Tor soro, Tor douronensis, and Neolissochilus sumatranus in the North Sumatra Rivers, has been conducted. The objective of this study was to determine the possible genetic divergence among congeneric fish and supply the genetic information for the design of conservation strategies. Fish specimens were collected from three rivers in North Sumatra Bahorok River (Langkat District), Asahan River (Toba Samosir District), and Batangtoru River (South Tapanuli District). The genetic analysis employed the Random Amplified Polymorphism DNA (RAPD) markers using a set of RAPD primers: OPA-2, OPA-3, OPA-6, OPA-7, OPA-9, OPA-11, and OPA-17. The amplified fragments were analyzed using the Numerical Taxonomy and Multivariate Analysis System (NTSYS) 2.00 program to construct the dendrogram of relationship among accessions. The genetic similarity coefficient among species of T. soro, T. douronensis, and N. sumatranus reached 0.44-0.86, with the lowest similarity at 44%. The cluster analysis indicated that T. douronensis is more closely related to N. sumatranus than T. soro based on the grouping of accessions. There is an indication of natural hybrids occurrence between N. sumatranus and T. soro populations despite the different habitats and locations of sampling. Our study revealed that using RAPD markers may discriminate inter- and intraspecific Batak fish populations in North Sumatra.

Genetic Analysis of (Oreochromis niloticus) Nirvana Tilapia Cultivated in Wanayasa and Galunggung, Lumajang, Bali, Minahasa Using Random Amplified Polymorphic DNA Method

This research aims to determine the kinship of nirvana tilapia (Oreochromis niloticus) cultivated in Wanayasa with Galunggung, Lumajang, Bali, and Minahasa by Random Amplified Polymorphic DNA method. All tilapia cultivated in these four places originated from Wanayasa. This research was conducted in the Biotechnology Laboratory, Laboratory of Microbiology and Molecular Biotechnology, FPIK UNPAD. The research method was an explorative experiment with the RAPD-PCR technique using primary OPA-3 and OPA-5. However, the results of DNA amplification with OPA-3 were better than with OPA-5. Therefore, further analysis was carried out using data from OPA-3. Kinship was analyzed based on the similarity index which was calculated by Numerical Taxonomy and Multivariate Analysis System (NTSYS) program. The results showed that the kinship between Nirvana tilapia cultivated in Wanayasa with Galunggung, Lumajang, Bali, and Minahasa, according to the similarity index was high, about 77-85%. The highest similarity index (85%) was obtained between nirvana tilapia cultivated in Wanayasa with Galunggung, and the lowest similarity index (77%) between tilapia cultivated in Bali with Minahasa. Genetic changes that occur because of differences in cultivation environments range from 15-23%.

Biodiversity of lactic acid bacteria of wild honey bee and Sumbawa horse milk by using RAPD-PCR

Wild honey bee and Sumbawa horse milk is the local product from West Nusa Tenggara, Indonesia, which has the characteristic of little sour in taste. The sour taste indicates lactic acid bacteria (LAB) existence that has a potential function as probiotics. The objection of this study is analysis of lactic acid bacteria biodiversity from wild honey bee and Sumbawa horse milk by using Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR). The genome of cultured samples from M17 and MRS medium were extracted, then identified by RAPD-PCR with primer (GTG)5 (5-GTGGTGGTGGTGGTG-3). The scoring of DNA fragments was analyzed using the Numerical Taxonomy and Multivariate Analysis System (NTSYS) application. The result show five clusters from a common ancestor indicated the broad diversity of LAB in wild honey bee and Sumbawa horse milk.

Morphological characteristics of Phaius spp. orchids from Indonesia

Abstract. Hartati S, Samanhudi, Manurung IR, Cahyono O. 2021. Morphological characteristics of Phaius spp. orchids from Indonesia. Biodiversitas 22: 1991-1995. Orchid is one of the ornamental plants with great economic value in various communities. Characterization and grouping of several orchid species can be used in breeding efforts to obtain hybrid with the desired traits. This study is aimed to characterize the morphological appearance of Phaius orchids and to cluster the species based on their similarity in morphological characteristics. Five species of Phaius (P. tankervilleae, P. indigoferus, P. montanus, P. amboinensis, and P. callosus) of the collection of the Bogor Botanical Garden were observed based on 30 morphological characters including the characters of stems, pseudobulbs, leaves and flowers. The data obtained were converted into binary data and computed using the Numerical Taxonomy and Multivariate Analysis System (NTSYS) Spc 2.02 program to produce dendrogram. The results show that based on the morphological characterization of the five species of Phaius spp., they have similarities from 63% to 80%. Among the five Phaius species, P. amboinensis, P. tankervilleae, and P. montanus had 80% similarity and can be used as parents for crossing.

Morphological characterization of Calophyllum Inophyllum as a biodiesel

Nyamplung (Calophyllum inophyllum Linn.) is one of the most potential forest plant species to be flourished. C. inophyllum seed can be deformed into oil as biofuel. Biofuel is very important to mitigate climate change impact as it reduces greenhouse gas emission from transportation sector. Mostly C. inophyllum grows along the coastline, especially in Ngombol and Grabag, Purworejo. There are a lot of C. inophyllum plant populations at these sites, but its varieties are not studied yet, therefore it is necessary to study the characterization of its morphology. This study aimed to obtain morphological character data and analyze C. inophyllum plants’ diversity in Purworejo. Samples were selected intentionally (purposive sampling). Thus morphological data elaborated descriptively and the cluster was analyzed by NTSYS (Numerical Taxonomy and Multivariate Analysis System) program. The study showed the diversity and also the similarity of morphological characters. C. inophyllum fruit and seed’s morphological character had its fruit length 2.1 – 3.1 cm, fruit diameter 1.93 – 1.95 cm, fruit weight 3.04 – 12.82 g, fruit shape is spherical/round, and fruit hardness 92.5 – 99.57 N. Moreover, it had seed length 1.16 – 2.22 cm, seed width 1.16 – 1.79 cm, seed weight 0.74 – 3.06 g, seed shape were spheroid, ellipsoid, reniform and the color was yellow. The morphological characters’ variance was 26%, which was categorized as low and Grabag district had more potential for growing and production than Ngombol regency.

Morphologic and cytoplasmic assessments of bulb onion (Allium cepa L.) landraces

Today climate change threatens to reduce crop yield and harming the food security. Local landraces have adaptation skills to shifting climatic conditions. Using of this local source in plant breeding programs becoming an alternative strategy. In this study, 97 landraces were collected to initiate the bulb onion breeding program eligible for the current trends. Collected materials were morphologically characterized using 21 descriptors, derived from UPOV (International Union for the Protection of New Varieties of Plants). Clustering which was conducted by the NTSYS (Numerical Taxonomy and Multivariate Analysis System) program using UPGMA (Unweighted Pair Group Method of Arithmetic Averages) method, showed that the genetic similarity rate of the landraces was calculated between 0.06-0.96. Hybrid onion breeding program depends on the cytoplasmic-genic male sterility (CMS) system. Thus, the PCR-markers were applied to identify the cytoplasm types of the landraces. Among landraces N-cytoplasm was found in 78 accessions and S-cytoplasm was found in 19 accessions. At the end of the study, a qualified gene pool has been established consisted of characterized onion genotypes which will might be used in further breeding studies.

Analysis of molecular characterizations of beets, Beta vulgaris in response to cyst nematodes, Heterodera schachtii

Sugar beet cyst nematode Heterodera schachtii is a serious biotic limiting factor in sugar beet industry. In this study, 70 sugar beet accessions were screened against H. schachtii, of which 30 resistant and susceptible ones were subjected to polymorphic simple sequence repeat (SSR) marker analysis by seven Universal DNA Amplification (UDA) primer pairs randomly. The results showed a very high significant variation between resistant and susceptible sugar beet accessions in response to H. schachtii with 42 fold increase in nematode reproduction rate in susceptible ones. SSR primers produced 155 bands, out of which 91 ones showed polymorphism, indicating 59% genetic diversity. The highest polymorphic bands was of UDA013 primer with 17 and the lowest one in UDA010 with 10 bands. Average number of polymorphic bands per primer was 13 and per genotype 3.03 bands. The UDA015 had the highest polymorphic index content (PIC) of 80%, and the lowest 44% in UDA001 with the mean of 67%. Highest marker index (MI) was in UDA015 with 10.80, and lowest one 6.48 in UDA001, with the mean of 8.68. Cluster analysis using Numerical Taxonomy and Multivariate Analysis System (NTSYS) software, with UPGMA method based on Jacard's similarity matrix, divided the genotypes into 4 major clusters. The dendrogram was consistent with the clustering result of Principal Component Analysis (PCoA). The highest genetic similarity of 91% was between the genotypes 45 and 51, of which both were offspring of a half-brother and half-sister crossing. The results suggested novel sources of resistant to H. schachtii and that SSR marker is a reliable tool for revealing a high levels of polymorphism among the sugar beet genotypes.

In silico design and in vitro validation of a novel PCR-RFLP assay for determination of phylogenetic clusters of streptokinase gene alleles in streptococci groups

Streptokinase (SK), a heterogeneous plasminogen (Pg) activator protein secreted by groups A, C and G streptococci (GAS/GCS/GGS) is a virulence factor composed of three structural domains; SKα/SKβ/SKγ. Phylogenetic analysis of the major variable region of SKβ (sk-V1; nucleotides 448–791; 343bp) which classifies the SK alleles into SK1/SK2 clusters and SK2a/SK2b sub-clusters, is an approved assay to categorize clinical/natural streptococcal-isolates into co-related functional/pathogenesis groups. Herein, we describe a novel PCR-RFLP assay that in combination with Numerical Taxonomy and multivariate analysis System (NTSYS) resulted to dendrograms with complete adaption to that of the phylogenetic analysis of sk-V1-based clustering. In silico analyses by 30 restriction enzymes on GenBank-acquired sk-V1 sequences of known streptococcal clusters, resulted to the selection of “BsrI, MseI and Tsp45I″ enzymes that produced proper patterns to construct the expected dendrograms. In vitro analysis of the selected enzymes on clinical isolates of GAS/GCS/GGS validated the production of the same in silico-observed digestion patterns. Comparison of the constructed dendrogram and phylogenetic trees of selected GenBank and clinical isolates of streptococci indicated complete adaptation. Assessment of Pg-activation activity in selected clinical isolates indicated the expected co-related functionalities of the classified SK-clusters by the invented PCR-RFLP/NTSYS method. The simplicity of the assay relieves the need of sequencing/phylogenetic analyses for SK-clustering.

Banana diversity (Musa spp.) as a result of in vitro gamma ray irradiation based on morphological markers.

Abstract. Due MS, Yunus A, Susilowati A. 2019. Banana diversity (Musa spp.) as a result of in vitro gamma ray irradiation based on morpological markers. Pros Sem Nas Masy Biodiv Indon 5: 347-352. Banana is a horticultural commodity that has a high enough production so that it makes it a superior product in the country. Banana has long been domesticated by the community because it has many benefits. There are more than 200 banana cultivars growing in various regions, one of which is feather plantain. Banana plants are propagated vegetatively so that they have limitations in obtaining genetic variation and require a long generation time in their vegetative cycle. Improvements in the nature of banana plants can be done by increasing its genetic diversity through mutation induction using gamma ray physical mutagens combined with in vitro culture techniques.This study aims to find out the diversity of feather plantain as a result of in vitro gamma ray radiation based on morphological markers. This research is a type of experimental research using completely randomized design (CRD) method. The factors tested were 4 radiation doses (0 Gy, 10 Gy, 20 Gy and 30 Gy). Morphological observations are carried out based on quantitative and qualitative characters. The data obtained were analyzed using the SPSS ver. 10 and the NTSYS (Numerical Taxonomy and Multivariate Analysis System) program ver. 16. The result of this study shows that gamma ray radiation in various doses causes the emergence of variations in plants, especially in the stems and leaves. Gamma ray radiation at a dose of 30 Gy is the most optimal dose to produce diversity.

The morphological diversity of Chrysanthemum resulted from gamma ray irradiation

Abstract. Susila E, Susilowati A, Yunus A. 2019. The morphological diversity of Chrysanthemum resulted from gamma ray irradiation. Biodiversitas 20: 463-467. Chrysanthemum is a popular ornamental plant. The high market demand makes breeders develop new cultivars to meet market preferences. One way to get superior varieties of Chrysanthemum is by using gamma ray mutation induction which will increase the morphological variations. The purpose of this study is to determine the Chrysanthemum’s morphological diversity resulted from gamma irradiation through morphological markers. This experimental research used a completely randomized design (CRD) with one factor, i.e. radiation doses: 0 Gy (control), 10 Gy, 15 Gy, and 20 Gy. The observations were carried out qualitatively and quantitatively. The data obtained were analyzed using the SPSS 15.0 and the NTSYS (Numerical Taxonomy and Multivariate Analysis System) 2.02 program. The results showed that increasing the dose of gamma irradiation inhibited plant growth. Irradiation at 10 Gy and 20 Gy produced the most color changes in flowers compared to other doses. The basic color of 0 Gy irradiation is purple. Irradiation at 10 Gy and 20 Gy produced dark purple and deep red. Quantitative data showed that the treatment of gamma ray irradiation significantly affected the leaf length, leaf width, stem diameter, stem length, and diameter of flowers. The irradiation also caused changes in shape and texture of the leaves. The dendrogram showed that 20 Gy irradiation resulted the most diverse morphology compared to the control, 10 Gy and 15 Gy.

Morphological and genetic analysis of batak fish (Tor soro) in North Sumatera

Research on the analysis of morphological and genetic variations of Batak fish (Tor soro) in North Sumatra has been conducted. The Batak fish samples were captured from flowing rivers at several districts in North Sumatera namely: Bahorok River (Langkat), Alian River (Toba Samosir), Batang Toru River (South Tapanuli). Truss method was used to analyze morphological parameters by using SPSS ver. 16.0 computer program, while the genetic parameters were analyzed by scoring amplified DNA strands visualized in electrophoresis using Polymerase Chain Reaction – Random Amplified Polymorphism DNA (PCR-RAPD) method. Primers used in this study were: OPA-02, OPA-03, OPA-04, OPC-01 and OPC-02. The PCR amplicons were then analyzed by using Numerical Taxonomy and Multivariate Analysis System (NTSYS) ver. 2.02. computer program. Dendrogram analysis showed that the level of morphological variability was at 1–25 euclidean distance. Dendogram analysis also showed that genetic diversity level was within coefficient of 0.23–0.71 in which obtained a diversity level of 48%. Based on the morphological and genetic results, it may be indicated that the Batak fishes originating from Batang Toru had a close similarity with Alian’s rather than Bahorok’s.

The Genetic Variations And Relationship of Madura Tobacco (Nicotiana tabacum L.) Based on Molecular Characteristics

Madura has at least 22 genotypes of local tobaccos (Nicotiana tabacum L.). This diversity could potentially produce new genotype of tobaccos with superior characters. However, information of the genetic diversity of Madura tobaccos is still limited. The aim of this study was to determine the genetic variation and relationship of 24 genotypes of Madura tobaccos with Random Amplified Polymorphic DNA (RAPD) analysis. In this research we were used 6 single primers for amplification: (OPA-18, OPB-12, OPB-14, OPC-1, OPC-8 and OPC-19) and 2 mixture primers ((OPB-12+OPC-8) and (OPC-1+OPC-19)). Genetic similarity and clustering was analyzed with Unweighted Pair Group Method Arithmetic (UPGMA) method with Numerical Taxonomy and Multivariate Analysis System (NTSYS) version 2.10 software. From this research we found that OPA18425, OPB12450, OPC8500, (OPC19+OPC1)550 and OPC8800 can be used as specific markers. Polymorphic bands percentage with mixture primers was relatively equal with single primers (<60%). The dendogram showed that Madura tobacco genotypes consist of 2 main clusters: cluster A (22 genotypes) and cluster B (2 genotypes: Bukabu Sa’ang and Prancak-95). Madura tobaccos had high genetic similarity between genotypes ranging from 0.80-1.00.

Morphological and genetic diversity of bamboos in Arboretum Universitas Padjadjaran, Sumedang, Jawa Barat

Annisa, Mardliyyah A, Kusmoro J. 2017. Morphological and genetic diversity of bamboos in Arboretum Universitas Padjadjaran, Sumedang, Jawa Barat. Pros Sem Nas Masy Biodiv Indon 3: 351-360. Bamboos are versatile plant that have been used for a long time by Indonesian people in everyday life. The utilization of bamboo, continuously, could possibly reduced species and population of bamboos that available in nature. The study of morphological and genetic diversity can assist in gathering data for the purpose of conservation and sustainability use of bamboo’s germplasm. Research aimed to identify morphological and genetic diversity of bamboos in Arboretum Universitas Padjadjaran. The research of morphological diversity was done by identification based on the result of morphological observation and vegetative organ morphometric, while the research of genetic diversity was done by using SSR molecular marker. This research used observation method which included morphological identification and determination of location for collecting plant materilas, collecting plant materials, DNA isolation, visualization of DNA products, and amplification by using SSR marker. Morphology and amplification data were analysed descriptively. Clustering analysis based on genetic was done by UPGMA on Numerical Taxonomy and Multivariate Analysis System (NTSYS) 2.02. Result of morphological diversity research showed there were ten species of bamboos in Arboretum Universitas Padjadjaran that consisted of four genus such as Bambusa, Gigantochloa, Phyllostachys, and Schizostachyum. Result of genetic diversity research showed that ten loci of SSR markers designed from Phyllostachys in China and Bambusa in India can be applied on Arboretum’s bamboos . The total alleles were 117 alleles with range size of 83-893bp. PIC value of the primers were ranged between 0.84-0.94, which categorized as very informative. Based on similarity coefficient values it can be concluded that bamboo Arboretum Universitas Padjadjaran has high level genetic diversity.

GENETIC VARIABILITY OF Rottboellia cochinchinensis POPULATIONS IN SUGARCANE FIELDS

ABSTRACT The hypothesis assumed was the existence of biotypes within populations, which has been the cause of difficulties in itchgrass control by farmers. For that, the genetic variability of three populations of Rottboellia cochinchinensis in sugarcane fields in the state of São Paulo was investigated by using the Amplified Fragment Length Polymorphism (AFLP) technique. Six primers were used to obtain molecular characterization data. AFLP gels were analyzed based on marker presence (1) and absence (0). Using NTSYs (Numerical Taxonomy and Multivariate Analysis System) software, the genetic similarity was calculated by the Jaccard coefficient and, from that, a dendrogram was built through the UPGMA (Unweighted Pair Group Method Arithmetic averages) method, besides determining the isopolymorphic marks. The average genetic similarities seen in the region was 0.742 for Igarapava, 0.793 for Mococa and 0.808 for Piracicaba. Between regions it was 0.730 (Igarapava vs Mococa), 0.735 (Mococa vs Piracicaba) and 0.694 (Igarapava vs Piracicaba). In line with the dendrogram, it is possible to detect the formation of two groups, one with 8 plants from Igarapava and Mococa and the other with 21 plants from Igarapava, Mococa and Piracicaba, as well as the presence of 1 discriminant individual from Piracicaba. It can be concluded that the genetic similarity among itchgrass populations from the state of São Paulo was high (72%), which denotes that the difficulties in chemical management are not only due to different biotypes but also due to other characteristics linked to tolerance of the species to herbicides. However, biotype existence cannot be discarded because of the polymorphic marks generating 22% average genetic variability.

Diversity of edible canna (Canna edulis Ker.) in Surakarta region based on the morphological characters and isozym band pattern.

Ashary SS, Etikawati N, Suratman. 2016. Diversity of edible canna (Canna edulis Ker.) in Surakarta region based on the morphological characters and isozym band pattern. Bioteknologi 13: 18-33. The aim of the research is to study the diversity and relationship of Edible Canna in Surakarta region based on the morphological characters and isozym band pattern. This research included morphological observation of Edible Canna and electrophoresis of the rhizomes to get isozym band pattern. Edible Canna’s fresh rhizomes were extracted with extract buffer and then centrifuged at 13000 rpm during 20 minutes. Supernatan and loading dye were mixed and then electrophored with poliacrilamide gel. The morphological characters and isozym band pattern were analysed using Similarity Index (SI) and computed with Numerical Taxonomy and Multivariate Analysis System (NTSYS) program 2.0 version until get the dendogram. From the research, it can be concluded that there were diversity of Edible Canna based on the morphological characters and isozym band pattern. The diversity of morphological ganyong included the colour and size of Edible Canna’s organs. The diversity of isozym band pattern included emergence and thickness of band. Based on the morphological characters and isozym band pattern, Edible Canna from Wonogiri and ganyong from Sukoharjo have the closest relationship with similarity coefficient 48%, whereas the farthest was the relationship of C. hybrida with the others at similarity coefficient of 11.57%.

Isozyme analysis based genetic fidelity assessment of micropropagated banana plants

Isozyme studies of micropropagated and mother plants of banana cvs. Matti, Ney Poovan, Kechulepa, Dwarf Cavendish, Malbhog, Champa, B.B. Battisa and FHIA-1 were done to test their genetic fidelity. The banding patterns as revealed by electrophoretic variations were evaluated with respect to isozymes of acid phosphatase, catalase, esterase and peroxidase as markers. The genetic fidelity of micropropagated plants and the relationship of the different cultivars were determined by dendrogram using numerical taxonomy and multivariate analysis system(NTSYS). A clustered dendrogram was prepared by unweighted pair group method using averages (UPGAMA) method. At 87% similarity, the micropropagated and mother plants were clustered in four groups reflecting their genomic constitution. Cvs. Matti (AA) and Dwarf Cavendish (AAA) with similar ‘A’ genome were categorized in Cluster I. Cluster II comprised of cvs. Ney Poovan (AB), B.B. Battisa (ABB) and FHIA-1(AAAB) with genomic constitution of both ‘A’ and ‘B’ type. Cvs. Champa (AAB) and Malbhog (AAB) with similar genome were grouped inCluster III. Cluster IV contained the cv. Kechulepa (BB) having only ‘B’ genome. However, there was no somaclonal variation among the micropropagated plants and they showed 100% genetic similarity. Thus, the isozyme studies could be a reliable marker for testing the genetic fidelity of micropropagated plants and for evaluating the diversity among the banana germplasm.

Genetic diversity analysis among male and female Jojoba genotypes employing gene targeted molecular markers, start codon targeted (SCoT) polymorphism and CAAT box-derived polymorphism (CBDP) markers

To detect genetic variations among different Simmondsia chinensis genotypes, two gene targeted markers, start codon targeted (SCoT) polymorphism and CAAT box-derived polymorphism (CBDP) were employed in terms of their informativeness and efficiency in analyzing genetic relationships among different genotypes. A total of 15 SCoT and 17 CBDP primers detected genetic polymorphism among 39 Jojoba genotypes (22 females and 17 males). Comparatively, CBDP markers proved to be more effective than SCoT markers in terms of percentage polymorphism as the former detecting an average of 53.4% and the latter as 49.4%. The Polymorphic information content (PIC) value and marker index (MI) of CBPD were 0.43 and 1.10, respectively which were higher than those of SCoT where the respective values of PIC and MI were 0.38 and 1.09. While comparing male and female genotype populations, the former showed higher variation in respect of polymorphic percentage and PIC, MI and Rp values over female populations. Nei's diversity (h) and Shannon index (I) were calculated for each genotype and found that the genotype “MS F” (in both markers) was highly diverse and genotypes “Q104 F” (SCoT) and “82–18 F” (CBDP) were least diverse among the female genotype populations. Among male genotypes, “32 M” (CBDP) and “MS M” (SCoT) revealed highest h and I values while “58-5 M” (both markers) was the least diverse. Jaccard's similarity co-efficient of SCoT markers ranged from 0.733 to 0.922 in female genotypes and 0.941 to 0.746 in male genotype population. Likewise, CBDP data analysis also revealed similarity ranging from 0.751 to 0.958 within female genotypes and 0.754 to 0.976 within male genotype populations thereby, indicating genetically diverse Jojoba population. Employing the NTSYS (Numerical taxonomy and multivariate analysis system) Version 2.1 software, both the markers generated dendrograms which revealed that all the Jojoba genotypes were clustered into two major groups, one group consisting of all female genotypes and another group comprising of all male genotypes. During the present investigation, CBDP markers proved more informative in studying genetic diversity among Jojoba. Such genetically diverse genotypes would thus be of great significance for breeding, management and conservation of elite (high yielding) Jojoba germplasm.

Indonesia

Permatasari, Pitoyo A, Suratman. 2014. Keragaman suweg (Amorphophallus campanulatus) di wilayah eks karesidenan Surakarta berdasarkan karakter morfologi, anatomi dan pola pita isozim. Bioteknologi 11: 11-18. The aims of this research were to find out the diversity and relationship among elephant yam in Surakarta region based on variation of morphological, anatomical and isozyme banding patterns characters. Total 20 samples were taken using random purposive sampling technique in different altitude of 7 districts. Morphological, anatomical and isozyme banding pattern data were analyzed descriptively. The steady data of quantitative characteristic from morphological, anatomical and isozyme banding pattern of binary data was analyzed using an Index of Similarity (IS) and computed by the program Numerical Taxonomy and Multivariate Analysis System (NTSYS) version 2.10. The result showed that the elephant yam in Surakarta region were diverse based on morphological, anatomical and isozyme banding pattern, especially in ratio and color of petiolus; tuber ratio; index of stomatal; thick epidermis; thick mesophyll; term and ratio of palisade; term rafida; 4 peroxidase and 5 esterase isozyme banding pattern. Analysis of relationship of elephant yam based on morphological, anatomical and isozyme band pattern characters showed the similarity coefficient of 64,85% contained two major group. The first group consist of Boyolali 1, Boyolali 3, Boyolali 4, Wonogiri 4, Wonogiri 2, Karanganyar 1, Karanganyar 2, Karanganyar 3, Sragen 2, while the other samples were separated from this group.

In vitro callogenesis and detection of somaclonal variations in Plantago ovata L.

Planago ovata L. is an economically important species in the monotypic genus Plantago. It is a short-stemmed annual herb. The seed husk of this plant is commonly called psyllium or isabgol which is important in pharmaceutical formulation and food industry. In this study, callus induction was optimized using different explants of Plantago ovata. Callus DNA was utilized to access the somaclonal variations using the Random Amplification of Polymorphic DNA (RAPD) markers. The maximum callus growth was observed in Murashige and Skoog (MS) medium containing 4 mg L−1 2,4-D concentration for shoots, 0.5 mg L−1 for seeds and 2 mg L−1 for roots. Moreover, the effect of culture age was considered in assessing genetic variability. Maximum genetic variability was observed in the DNA samples of callus at the concentration of 2 mg L−1 2,4-D for all explants (roots, shoots, and seeds). Cluster analysis was performed based on 1) similarity coefficient between samples and 2) molecular data using the Numerical Taxonomy and Multivariate Analysis System (NTSYS) PC version 2.01; similarity index was generated by similarity for Quantitative Data (SIMQUAL). Our study indicated that Random Amplified Polymorphic DNAs can successfully be used to explore polymorphism among callus samples at different hormonal concentrations. This study can be useful for the production of callus from Plantago ovata and estimation of genetic variations due to tissue culture conditions. Evaluation of genetic variations can display novel features and manipulate genetic bottlenecks in Plantago ovata. New genetic variations in somaclones can bring vital insight for plant improvement.

Phylogenetics of selected Mentha species on the basis of rps8, rps11 and rps14 chloroplast genes

Mentha is a genus of family Lamiaecae, and is well known for its great medicinal and economic values. It is widely distributed over five continents (excluding Antarctica and South America) of the world. In order to construct the phylogeny and to investigate the genetic variability among seven Mentha species polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (CAPS) marker technique was used. Three chloroplast genes rps8, rps11 and rps14 were used to amplify from the chloroplast genome of sevenMentha species. rps8 gene was tested on broad range of annealing temperatures but no amplification was observed while rps11 and rps14 regions of Mentha cpDNA were successfully amplified and subjected to PCR-RFLP. For restriction digestion of the amplified PCR product, twelve different restriction enzymes were used and the resulting restriction pattern was resolved on PAGE. Comparison of Nicotiana tabacum and Mentha rps11 andrps14 genes was also performed. The data was analyzed by using the software numerical taxonomy and multivariate analysis system (NTSYS) pc version 2.01 showing considerable level of genetic similarity among the Mentha species. It was also found that Eam1104I andAlw44I based PCR-RFLP of rps14 gene could serve as a specific marker for Mentha spicataidentification and differentiation. Key words: Mentha, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), chloroplast genes, rps, phylogenetic analysis.