Research Journal of Biotechnology

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Vol. 8(12) December 2013

Extraneous endophytic Bacteria Enhanced Rooting and growth of Liquidambar formosana In vitro

Lina Xu, Minsheng Yang*, Dietrich Ewald and Akindele Akinnagbe

Endophytic bacteria can promote the rooting and growing of plants. In this paper, Paenibacillus (P22) and Stenotrophomonas (S16) isolates from hybrid poplar were used to inoculate shoots of Liquidambar formosana in order to investigate the effects of P22 and S16 on the rooting and growth of the plants. The result showed that P22 is stronger than S16 in enhancing the rooting and growth of tissue-cultured Liquidambar formosana and both bacteria equally proved to be plant-growth-promoting bacteria that have capability to grow on a medium with low nitrogen content. Thus, the result demonstrated the possibility of isolating plant-growth-promoting bacteria from its natural host to promote the growth of another plant.

Effects of Abscisic acid (ABA) carotenogenesis expression and astaxanthin accumulation in Haematococcus pluvialis

Gao Zhengquan, Gao Hongzheng and Meng Chunxiao*

Haematococcus pluvialis is a commercially promising source because of its ability to accumulate astaxanthin up to 4% (w/w) of dry weight under stress conditions. In this study, transcriptional expression pattern of eight carotenoid genes involved in astaxanthin biosynthesis under ABA (25 and 50mg/L) induction was investigated using qRT-PCR. Results showed that except for bkt, almost all genes were up-regulated by ABA with different expression profiles. ABA25 induction had a greater effect on the transcriptional expression level of ipi-1, ipi-2, lyc and crtR-B than on psy, pds and bkt. ABA50 treatment had a greater impact on the transcriptional expression of ipi-1, ipi-2 and crtR-B than on pds, lyc, bkt and crtO. Moreover, astaxanthin biosynthesis under ABA induction might be up-regulated by ipi-1, ipi-2, pds at transcriptional level; lyc performed at both transcriptional and post-transcriptional levels and crtR-b functioned at post-transcriptional levels. Furthermore, there seemed to be little effect on crtO transcriptional expression in ABA25 treatment and on psy mRNA expression in ABA50 treatment and both ABA treatments had no impact on mRNA expression of bkt.

Optimization of lipase production by Acinetobacter johnsonii G23

Dai Yujie*, Zhang Yehong, Sun Guojuan, Li Xiaolu, Wang Haikuan and Zhang Liming

Structural and sequence analyses of the enzyme require a certain amount of protein with high purity. However, the medium composition has an important effect on the production and purification of an enzyme. In this study, the production of a low temperature alkaline lipase by Acinetobacter johnsonii G23 was substantially enhanced through adjusting carbon and nitrogen sources and optimizing the concentration of inorganic salt. The result showed that a maximum lipase production of 13.6 U/ml was reached with the optimum culture medium containing 1% (w/v) soluble starch, 1.5% (w/v) soy peptone, 0.04%(w/v) MgSO4, 0.05%(w/v) CaCl2 and 1%(v/v) olive oil. With this condition of fermentation, clear and bright protein bands were obtained under a clean background in SDS-PAGE gel. This medium composition favors the further purification and sequence analysis of this lipase.

Antidiabetic activity of aqueous and alcoholic leaf extract of Paederiafoetida Linn. in streptozotocin–nicotinamide induced diabetic rats

Shetti Sandeep, Sanghai Dhirendra, Aswatha Ram H. N. and Shreedhara C. S.*

The antidiabetic potential of the leaves extracts of Paederia foetida Linn. (Rubiaceae), a medicinal plant widely used as the traditional folk medicine by Sikkim and Darjeeling Himalayan tribes, for the treatment of diabetes mellitus was evaluated in the STZ-nicotinamide induced type 2 diabetic model. Graded doses of the alcoholic and aqueous leaf extracts were administered to normal and experimental diabetic rats for 21 days. Fasting plasma glucose levels, serum insulin levels, serum lipid profiles and changes in body weight were evaluated in normal rats with no significant changes in these parameter, while liver glycogen levels, glycosylated hemoglobin and pancreatic TBARS levels were evaluated additionally in diabetic rats and were compared with diabetic control animals. The diabetic groups treated with the leaf extracts were compared with standard glibenclamide. Significant results were observed in the estimated parameters, thereby, justifying the use of the plant in the indigenous system of medicine.The findings of the study support the antidiabetic claims of Paederia foetida.

Effect of Mesoporous Silica Nanoparticles in the biodegradation of Polyoxymethylene polymer

Dhanalakshmi K. G.* and Renuka Devi P.

The engineering thermoplastic polyoxymethylene (POM) is a polymer of formaldehyde. Even though it is believed to replace the usage of metals, it also degrades easily under many circumstances like heat, acids, alkalis, oxygen, abrasion, enzymes, radiations etc. This work aims to study the degradation of POM Nano composite with Mesoporous Silica Nanoparticle (MSN) and pure POM at periodical intervals. Degradation in two different atmospheres viz., natural withering and soil buried condition were studied. The POM materials were characterized with Tensile strength, Diffused Reflectance Spectroscopy (DRS) and Fourier Transform Infra Red Spectroscopy (FTIR), before and after degradation. These characterizations revealed that tensile strength of the pure POM material was lost significantly after six months of natural withering whereas POM nanocomposite with MSN showed no significant loss, indicating stability of the material due to MSN. Both DRS and FTIR supported this result. By DRS, it was identified that the optical property of the materials increased due to degradation. FTIR shows differences in the peak intensity for the aldehyde group after degradation. In all the cases soil buried samples showed quick degradation. Hence, microbial analysis was done with soil buried POM samples. Physical, biochemical and molecular characterization confirmed that the isolated organism was Methylobacterium sp.

Enhancement of Hydrocarbon Degrading and Biosurfactant Production of Pseudomonas aeruginosa Strain Z41 by Joint Employment of Low Energy Ion Beam and Ultraviolet Irradiations

Zhang X. S.

A biosurfactant producing oil degrading Pseudomonas aeruginosa strain Z41 was treated subsequently by low energy ion beam (LEIB) and ultraviolet (UV) irradiation for genetic improvement in crude oil removal efficiency as well as biosurfactant production. The cell suspension was first irradiated by 20 keV nitrogen ion beam at the fluence of 0-120×2.5×1013ions/cm2. The ion implantation fluence dependent survival curve showed “saddle” type as reported in literature. By assaying of the survival rates and mutation rates caused by ion beam implantation, the optimal fluence of ion implantation to mutate Z41 was determined at 60×2.5×1013ions/cm2, the inflection point of survival curve. One mutant named L18 was preliminarily screened out by alternative blood NA plate and oil NA plate screening. Then L18 was irradiated by with 20W violet lamp for 5s, 15s, 30s, 40s and 60s and subjected to similar screening. And one mutant, M23, was selected ultimately whose degradation rate was 52.1%, increasing by 29.6% compared to L18 and whose fermentation yield of rhamnolipid yield by shaking flask fermentation reached to 19.6 g/L, an increase by 28.9%. It was suggested that the joint employment of both the irradiations should make good results in mutation breeding.

Chemical dissolving of citric acid on bioleaching of copper mine tailings by Acidithiobacillus ferrooxidans ATCC 23270

Dou Li, Yajie Yin, Binquan Jiao*, Liyan Song, Yangqing Wang, Jing Qiao and Zhonghui Xu

Bioleaching could be a lower energy consumption and environmental friendly technology to recover precious metals from copper mine tailings and reduce its environmental risk. The citric acid is ubiquitous in copper mine tailings and its effects on bioleaching of copper mine tailings are unknown. The purpose of this study was to assess influence of the agents on bioleaching effect and estimating the mechanism. In this work, bioleaching of copper mine tailings using Acidithiobacillus ferrooxidans ATCC 23270 under aerobic conditions was conducted with addition of citric acid of different concentrations. The results from the detection of Cu, Mn, As, total Fe, ferric, ferrous, pH, Eh and cell numbers in leachate during the 30 days showed that the citric acid accelerated the metals leaching. The scanning electron microscope (SEM) images demonstrated the aggregates and tracks of acid attack that could prove the bioleaching results on the other hand and the chemical dissolving played an important role in the bioleaching process.

The effects of Changing Growth Conditions on the Demethylation of a Lignin-Related Compound by Absidiacylindrospora,Fomesfomentarius, Ischnodermaresinosum and Cylindrocladiumcamelliae

Gibson A., Grochowski J., Malek L. and Dekker R.F.H.

Culture parameters influencing the demethylation of the lignin-related compoundpara-vanillin in defined media were examined in culture flasks of wood degrading Absidiacylindrospora, Fomesfomentarius, Ischnodermaresinosum and Cylindrocladiumcamelliae with respect to methanol release, vicinal diol generation and protein content. Cultures were grown in 40 mL media containing 2% VMS, 1% glucose and demethylationactivity induced with 1 mL of400 mMpara-vanillin. Study of the effect of incubation and induction time indicating longer incubation length did not significantly affect demethylation. Vanillin inhibited fungal growth. Culture agitation at 81 rpm increased methanol released in Absidiacylindrospora and Fomesfomentarius by 27% and 87% respectively. The concentration of vicinal diols, however, decreased in Absidiacylindrosporaby 53% with 0-3 mM of hydrogen peroxide added, indicating hydrogen peroxide may have acted as a mediator in vanillin degradation. Changes in medium composition with respect to metals had 6-fold decrease in vicinal diol concentration for Fomesfomentarius with 0-2 μM final concentration of copper and 166% increase in methanol concentration for Cylindrocladiumcamelliae with 0-0.5 μM final concentration of iron. Control of culture conditions is essential for optimal lignolytic activity and will facilitate finding more reactive lignin useful in future industrial applications.

Studies on the antibacterial peptide isolated from Murrya Konigii leaves

Sabiha Imran

The aim of the present study is to evaluate the antimicrobial activity of aqueous extract of Murraya koinigii leaves against the gram positive and gram negative bacterial strains by observing the zone of inhibition. The gram bacteria used in the test were Staphylococcus aureus, Bacillus cereus and Micrococcus luteus and gram negative bacteria were E coli and Klebsiella pneumoniae. Extract of leaves after purification by ammonium sulphate and gel filtration chromatography showed varying degree of inhibition on the growth of tested microorganism. Results obtained after various experiments confirmed that antibacterial component was protein. This anti microbial peptide could be used as a potential drug candidate for the treatment of various pathogenic diseases without any side effects.

Design and Implementation of wide depth treatment pulse using Transcranial Magnetic Stimulation

Kim Whi-Young* and Kim Jun-Il

The transcranial magnetic stimulation recharges the energy storing condenser and sends the stored energy in the condenser to the pulse shaping circuit which then delivers it to the stimulating coil. Previous types of transcranial magnetic stimulation required a booster transformer, secondary rectifier for high voltages and a condenser for smooth type. The energy storing condenser is recharged by switching the high-voltage direct current power. Loss occurs due to the resistance in the recharging circuit and the single-pulse output energy in the transcranial magnetic stimulation can be changed because the recharging voltage cannot be adjusted. In this study a booster transformer, which decreases the volume and weight, was not used. Instead, a current resonance inverter was applied to cut down the switching loss. A transcranial magnetic stimulation, which can simultaneously alter the recharging voltage and pulse repeats, was used to examine the output characteristics.

Genetic variants of milk protein at Romanian Grey Steppe breed beeing in preservation

Creangă Şteofil, Maciuc Vasile* and Bâlteanu Valentin Adrian

The paper presents a part of the results obtained in a complex research project on Romanian Grey Steppe breed, included in the genetic fund (G.D. no. 822/2008.) from Romania. For the Romanian Grey Steppe breed, we identified alleles for the six loci codifying the six types of major milk proteins alpha-casein S1 (α S1-cz); beta-casein (β-cz); kappa-casein (K-cz); beta-lactoglobulin (β-lg); alpha-lacto albumin (α-la); alpha-casein (α S2-cz). Milk analysis by I.E.F. allowed for the identification of a new allele in the loci of αS1-casein, for two cows from the total number of analysed animals (30 capita) from S.C.D.C.B. Dancu, Iaşi, Romania, an allele called αS1-casein IRV. It has the shape of a stripe with isoelectric point between allele B and C, closer to the one of allele C and two heterozygote individuals were identified as carrying these so-called alleles BIRV and CIRV. The presence of this ancestral allele common to the Romanian Grey Steppe breed shows the oldness of the breed and it is a premiere for the Podolic family. So, this new casein could be an important genetic marker for the breed. It could be an alarm signal for the intensification of the efforts to preserve the Romanian Grey Steppe breed.

Protective effects of Coriander extract against Puffer fish (Lagocephalus lunaris) induced toxicity

Mandal N. and Khora S. S.*

Coriander has a long history of use in traditional medicine. The objective of this study is to evaluate the protective effects of an aqueous extract (5 g/l) of C. sativum (leaves and stems) on biochemical disorder, oxidative stress and histopathological changes induced by muscle extract of puffer fish Lagocephalus lunaris in Swiss albino mice. Mice injected with L. lunaris muscle extract (1 ml/100g body weight) for 10 days showed (1) a reduced appetite and diarrhea resulting in a lower growth rate than controls, (2) an increase in serum Alanine aminotransferase, Aspartate aminotransferase, Alkaline phosphatase and Total and Direct bilirubin levels suggesting liver functional disorders, (3) an increase of serum creatinine and uric acid concentrations highlighting renal insufficiency and (4) increase in the level of serum triglycerides, total cholesterol, glucose and low density lipoproteins indicate the interference in the metabolism or biosynthesis of lipids (5) an oxidative stress as evidenced by the raise of TBARS and the inhibition of antioxidant enzymes (Superoxide dismutase, Catalase and Glutathione peroxidase) activities in liver, kidney and heart tissues (6) histopathological changes in liver, kidney and heart tissues. Absorption of C. sativum aqueous extract as a drink, for 10 days demonstrated ameliorative action and prevented the disorders induced by L. lunaris muscle extract.

Staphylococcus epidermidis in Human Skin Microbiome associated with Acne: A Cause of Disease or Defence?

Pathak Rajiv, Kasama Nitesh, Kumar Raj and Gautam Hemant K.*

Propionibacterium acne is a well known bacterium having major role in acne development. However, the role of other skin microbiome in acne development/prevention is still not clear. One of the major problems was associated with poorly understood acne-associated uncultured bacterial populations, influencing acne directly or indirectly. With this in mind, we sought to study the remaining bacterial populations associated with acne and tried to investigate the role of these acne-associated human skin microbiome in disease induction as well as in its prevention. In order to identify the un-explored microbial populations present in acne patients, human skin microbiome isolated from acne lesions were investigated via culturable as well as metagenomics approaches and then the quantitative analysis of two most abundant microbial populations, P. acne and Staphylococcus epidermidis was performed using 16S rRNA gene expression analysis. The populations of P. acne and S. epidermidis were increased in acne patients, as compared to control in a very specific pattern. To understand the overall relevance in terms of biological function, skin explant models were used, in which skin was infected with P. acne, Staphylococcus capitis and S. epidermidis along with possible combinations. Interestingly, we observed that S. epidermidis was involved in acne prevention and an induction of antimicrobial activity was also observed in S. epidermidis after strain improvement during gamma irradiation exposure. It was tempting to speculate that S. epidermidis, a part of the human skin microflora is having a role in acne prevention whereas P. acne and S. capitis are having role in disease progression.

Agrobacterium-mediated transformation of millet [Setaria italica (L.)] using mature seeds

Jincheng Yuan, Huiping Wei, Yinghui Liu* and Zhiping Dong*

In this paper we report a simple protocol for Agrobacterium-mediated transformation system of millet. Two millet genotypes Jigu11 and Yugu2 and their mature seeds are used as receptor.The Agrobacterium strain EHA105 harboring with binary vector pCAMBIA1301, which contained hptII as select marker gene and GUS as reporter gene, is used for optimization of transformation conditions. The optimal conditions for the Agrobacterium mediated transformation are found to be wound and vacuum treat prior inoculum, exposure of explants for 30 min to Agrobacterial and 3 days of co-cultivation placed on medium supplemented with 200 uM acetosyringone (AS). Addition of 100 uM L-cysteine in the medium enhanced the frequency of transformation.

Co-grafts of Artificial dermis matrix and Autogenetic Split-thickness Skin repaired severe Hand Wounds of Deeply Burnt Patients

Hongqi Liu*, Li Yan and Deqian Sha

We evaluated the effect of using artificial dermis matrix plus autologous split-thickness skin (ADM and ASTS) in treatment of deep burn hands in large burned patients. A total of 58 large area burns patients were recruited, the burn areas were greater than 80% and have been Escher excised. Twenty eight of them were treated with ADM and ASTS as test group and 30 patients treated with autologous medium-thickness skin (AMTS) as control group. Jebsen–Taylor hand function test was compared and analyzed in two groups. The wound healing time in the test group (24.22±3.34 days) were longer than that of the control group (13.42±3.36 days) with statistical difference while the healing time of skin graft donor sites was shorter than control group (7.14±1.63 vs. 14.28±2.37 days) with statistical differences (P<0.05). The 3rd and 6th month follow-up results showed that clinical and functional evaluations have no difference in two groups and no obvious scar formation and less pigmentation in both groups resulted. Repairing of deeply burned hands with artificial dermis matrix was beneficial to both wound healing and donor site which was also beneficial to the rehabilitation of whole body for large burned patients.

Genotype independent tissue culture base line for high regeneration of japonica and indica rice

Sah Saroj Kumar* and Kaur Ajinder

The present investigation deals with genotype independent tissue culture base line for high regeneration of japonica and indica rice. The highest frequency of callus induction was observed in both the varieties on MS medium supplemented with 2,4-D (3.0 mg/L) + BAP (0.25 mg/L) + proline (600 mg/L) + maltose (40 g/L) + phytagel (3g/L). Embryogenic calli was subcultured on shoot regeneration MS medium supplemented with BAP (4.0 mg/L) + NAA (0.2 mg/L) + sucrose (30 g/L) + phytagel (2 g/L) + agar (8 g/L). In this medium regeneration percentage achieved was maximum. The rooting was carried out on half MS medium. The plants were hardened and transferred to soil in earthen pots. The developed method was highly reproducible. The developed plants showed normal growth and flowering under glass house conditions as well as in field conditions. To conclude tissue culture base line has been established in two varieties PR118 and Kitaake. Using this baseline, a total of 54 PCR positive transgenic lines were developed. The protocol developed here is genotype independent and is suitable for japonica as well as indica varieties.

The effects of JA treatment on the growth and some enzyme activities of eggplant embryos grown in vitro under salt stress conditions

Talhouni Manar*, Günalp Banu, Yaşar Fikret, Kuşvuran Şebnem, Uzal Özlem and Ellialtıoğlu Şebnem

In many areas of the world, soil salinization is an important concern for the sustainability of irrigated agriculture and accounts for large decreases in the yield of a wide variety of crops. Numerous attempts have been employed to optimize plant growth under saline conditions. One of them is to produce salt tolerant genotypes in different crops by traditional breeding. In addition, many other attempts have been made to overcome this disorder by exogenous application of plant growth regulators. Plant hormones are active members of the signal compounds involved in the induction of plant stress responses. Jasmonic acid (JA), which is naturally synthesized by plants, plays an important role as a signal molecule that induces the tolerance mechanisms under the influence of both biotic and abiotic stresses. In this study we aimed to investigate the effects of JA individually and/or accompanied with salt stress on two cultivars of eggplant embryos cultured in vitro. Embryos gently isolated from the 32-days old eggplant seeds and then were transplanted onto hormone-free MS medium basically containing 2% saccharose, 0.7% agar at pH 5.7. Different concentrations of NaCl and JA were applied with nine different applications. Two cultivars of eggplant were used: cv. Mardin Kızıltepe (M: Tolerant) and cv. Kemer (K: Sensitive). Germination percentage and antioxidative enzymes, SOD, CAT and APX activities were determined after harvesting. Among all applications of JA, plantlets that were pre-treated with 10 µM JA and then transferred to the salty media showed relatively better development than the other applications. Germination rate was 100% and plant growth was normal in control application. However germination rate and plant growth were inhibited by salt or JA application. The addition of JA caused more inhibition than salt stress. Among all applications of JA, embryos that were pre-treated with JA (cultured on the medium with 10 µM JA for 4 days and then transferred to the growth media with salt) showed relatively better development than the others. Antioxidative enzyme (SOD, CAT and APX) activities, increased with salt or JA treatments compared to control plants. The SOD, CAT and APX activities of the salt-tolerant Mardin Kızıltepe genotype were higher than salt-sensitive Kemer variety.

Comparative efficiency for genetic analysis of wild and cultivated germplasm of pigeonpea through RAPD and SSR markers

Walunjkar Babasaheb1, Parihar Akarsh*, Chaurasia Pratibha3 and Chauhan R. M.

The reliability of the quantification of genetic diversity using only one type of marker has been questioned as compared to the combined use of different markers. To compare the efficiency of the use of single versus multiple markers, the genetic diversity was quantified among 12 diverse pigeonpea germplasm comprising of eight wild and four cultivated using both RAPD and SSR markers. How well these two types of markers discriminated the diverse pigeonpea germplasm was evaluated. The pigeonpea germplasm including eight wild species and four cultivated varieties was subjected to 40 RAPD and 40 microsatellite primers. The level of polymorphism as revealed by RAPD primers produced a total of 517 DNA fragments and all were found to be polymorphic i.e. 100% and in SSR analysis 101 fragments were produced that too showed 100% polymorphism. The high similarity index value revealed by RAPD was 0.931 between GT-100 and ICPL-87 where as through SSR, it was 1.00 between GTH-1 and GT-100 as well as Rhyncosia rothi and R. minima. The least similarity index values revealed by RAPD (Rhyncosia rothi and GTH-1) and SSR (Rhyncosia bracteata and ICPL-87) were 0.07 and 0.133 respectively. Average PIC values showed by RAPD and SSR were 0.90 and 0.18 respectively. Using RAPD marker, the calculated arithmetic mean heterozygosity and the marker index were 0.90 and 22.47 respectively. The Rhyncosia bracteata and ICPL-87 were found distinct from rest of other cultivars by showing only 13% similarity. Average PIC value shown by RAPD and SSR primers were found to be 0.90 and 0.18 respectively.

Effects of fruit bagging on capsanthin and expression of key genes in the capsanthin biosynthetic pathway during fruit development and ripening of the pepper

Tian Shi-Lin, Gong Zhen-Hui*, Du Xiao-Hua and Shah Syed Noor Muhammad

Fruit bagging treatments have been widely used to improve the commercial value of fruit and to reduce agricultural chemical residue and pest and mechanical damage of fruit. However, bagging might also affect fruit quality. So, we used Cv.B54 as a model to determine the effects of fruit bagging treatments on capsanthin in the pepper and to understand the genetic regulation of capsanthin synthesis during fruit bagging treatments. Our results showed that bagging inhibited capsanthin synthesis during fruit development and ripening of the pepper; of note, bagging had the greatest effect on capsanthin during the mature-green stage of the fruit. Bagging reduced the accumulation of capsanthin and the mechanism behind this effect was that bagging inhibited the expression of key genes in the biosynthetic pathway of capsanthin. Most strikingly, this process inhibited Crtz and Ccs genes expression over the entire course of pepper fruit development and ripening, it may be a reason for capsanthin synthesis reducing. The results obtained indicate that choosing a kind of bags and an appropriate period of bagging are very important. In brief, this research has increased our understanding of the mechanism of capsanthin metabolism and has assisted in pepper production.

Dietary Review and Lipid Profile of Middle Aged Indians with special reference to Lipoprotein (a) levels and Risk Assessment for Cardiovascular Diseases

Mathur Ritu and Mathur Manika*

Cardiovascular diseases (CVD) account for high morbidity all over the world; risk factors include age, sex, hypertension, smoking, diabetes, high LDL and low HDL cholesterol levels. Elevated Lipoprotein (a) is an emerging independent risk factor in the development of cardiovascular diseases. Biochemical analysis revealed that 62.5 % of the subjects had elevated Lp(a) levels and 75 % of the subjects had elevated Homocysteine levels indicating their being at higher risk of CVD. Increased knowledge of the role of Lp(a) as a risk factor for CHD would be of great benefit. Because Lp(a) is genetically determined, we also recommend further studies to examine the relationship between family history of CHD and Lp(a) levels. Homocysteine levels in all the subjects were also found to be high. Dietary survey revealed a high intake of total fat, low intake of fiber and the P/S ratio indicated a higher consumption of saturated fats. Reducing serum homocysteine levels can reduce the risk of CVD which can be achieved by increasing the consumption of folic acid. Thus high risk subjects need nutrition education to control these risk factors for the prevention of this major disease.

Plant regeneration and corm formation of saffron (Crocus sativus L.) in vitro

Cavusoglu A.*, Sulusoglu M. and Erkal S.

In vitro plant regeneration and daughter corm formation of saffron (Crocus sativus L.) from corm parts consisting of meristematic region via direct organogenesis were performed. In the first experiment, corm explants were treated with five different sterilization procedures in order to overcome contamination difficulties as being encountered in all the geophyte. In the second experiment on direct shoot regeneration and providing the foliation, sterile meristematic node-containing corm explants were cultured on ½ Murashige and Skoog (MS) Medium or MS with several dose of BAP. According to the data MS+6 mg/l BAP had the best results in shoot initiation (96.7 %) and the foliation rate (93.3 %) while MS+1 mg/l BAP gave the least result in shoot initiation (16.7 %); MS+1 mg/l BAP and MS+10 mg/l BAP gave the least result in foliation from the initiated shoots in 90 days. In the third experiment, the daughter corm formation and rooting were achived on MS supplemented with IBA or IAA. The data showed that MS+1 mg/l IAA have the best results on daughter corm formation rate (76.7 %) and daughter corm number per corm formed explants (1.74 corms/corm formed explants). On the other hand rooting rate (46.7 %) and root number per root formed explant (1.5 roots/ root formed explant) were highest on MS with 2 mg/l IBA in 120 days. The protocol reported in this study may help in the growth and manupulation of agriculture and biotechnology of saffron which is increasingly conspicuous and demanding species.

Review Paper:

Pharmacological Relevance of Primitive Leafy Cactuses Pereskia

Sharif K. M., Rahman M. M.*, Zaidul I. S. M., Jannatul A., Akanda M. J. H., Mohamed A. and Shamsudin S. H.

Pereskia is the only cactus genus that has green leaf unlike other genera of large Cactaceae family. Pereskia is well known in scientific community for their heterogeneous, morphological and botanical characteristics. Although Pereskia genus originates in Latin American countries but it is also widespread in tropical areas like Malaysia, Indonesia. Some species of Pereskia have been used by traditional practitioners for the treatment of many diseases such as cancer, high blood pressure, diabetes, gastric pain and ulcer. Some of the species of this genus have been proven to possess diverse biological activities including anti-inflammatory, anti-microbial, antifungal, anti-nociceptive, anti-oxident and anti-cancer activities by several scientific investigations. The objective of this paper is to review the pharmacological activity and phytochemical properties of the Pereskia genus.