2(3) 2009 May issue
Efficient in vitro plant regeneration, flowering and fruiting of dwarf Tomato cv. Micro-Msk
Praveen Mamidala and Rama Swamy Nanna
Successful in vitro plant regeneration, flowering and fruiting were developed in Tomato (Micro-MsK) a model plant for genetic studies using leaf explants. The leaf explants were cultured on Murashige and Skoog’s medium (MS) supplemented with different concentrations (1.0-3.0 mg/L) of Benzyl amino purine (BAP) and Zeatin (Zt) individually and also in combination with 0.1 mg/L auxins (IAA/NAA). More number of multiple shoots (15.8) formation per explant was found at 2.0 mg/L Zt. Whereas maximum frequency of adventitious shoots were developed from the leaf explants cultured on MS medium augmented with 0.1 mg/L IAA + 2.0 mg/L Zt in comparison to all the concentrations and combinations used. To know the effect of Timentin on regeneration ability, leaf explants were also cultured on Timentin (100 - 400 mg/L) supplemented with IAA (0.1 mg/l) + Zt (2.0 mg/l) combination. High frequency number of multiple shoots followed by in vitro flowering and fruiting were observed. Viable seeds were formed and also showed the normal germination. The protocol developed in the present investigation may significantly contribute to genetic improvement of tomato.
Keywords: Tomato;Timentin; in vitro regeneration; flowering; fruiting.
Pages 98-102 Full Text PDF
Extraction of PCR-usable DNA from trees adapted to arid environment
G Sablok, P Gahlot, A.K.Gupta, K Pareek and N.S.Shekhawat
Genetic conservation programs in arid natural repertoire rely on molecular methods for diversity assessments. Molecular characterization involves the use of high molecular weight genomic DNA as starting material. Obtaining intact genomic DNA of sufficiently high quality, readily amplifiable using PCR is a primary goal of all molecular genetic studies. The aim of our study was to devise a method for the isolation of the genomic DNA from arid tree species viz. Prosopis cineraria, Calligonum polygonoides and Acacia nilotica (which are rich in polysaccharides, polyphenols and secondary metabolites), hold promises for the restoration of the region. The quality checking of the isolated DNA samples was done through the 10-mer oligonucleotide primers. The present method involves a modification of the available CTAB method employing higher concentration (6%) of polyvinylpyrrolidone (PVP), an extrinsic factor for reducing the metabolite precipitation, higher concentration of CTAB (3%) and an increased period (45 minutes) of incubation with chloroform: isoamylalcohol followed by the RNAse treatment for 90 minutes at 37°C and subsequent pelleting in TE buffer. The yield of DNA ranged from 1240-1500ng/µl and the absorbance lies between 1.7-1.9, indicating minimal levels of contaminating metabolites. The protocol has been tested with three tree species of the arid region which are extremely drought resistant. The DNA isolated was successfully amplified by all the random 10-mer oligonucleotide primers tested with high reproducibility. Optimum annealing temperature was 35°C at a concentration of MgCl2 (1.5mM), lower concentrations of primer (0.2µM) and Taq polymerase (1U). The present method is simple, efficient and economically yielding good quality intact genomic DNA suitable for large genetic screening programs.
Keywords: Prosopis cineraria; Calligonum polygonoides; Acacia nilotica; polyphenols; polysaccharides; DNA extraction; PCR.
Pages 103-109 Full Text PDF
Proteomics profile of pre-harvest sprouting wheat by using MALDI-TOF Mass Spectrometry
Abu Hena Mostafa Kamal, Ki- Hyun Kim, Dong- Hoon Shin, Hyung-Seok Seo, Kwang-Hyun Shin, Cheol-Soo Park, Hwa-Young Heo and Sun-Hee Woo
Wheat seed proteins were studied to identify the cultivar-specific proteins using two Korean pre-harvest sprouting wheat cultivars; Jinpum (susceptible) and Keumgang (resistant). Wheat seed proteins were separated by two-dimensional electrophoresis with IEF gels over pH ranges: pH 3.5-10. A total of 73 spots were digested with trypsin resulting peptide fragmentation were analyzed by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/MS). Mass spectra were automatically processed and searched through NCBInr, SWISS-PORT and MSDB database with mono isotopic masses. These proteins profiles are divided into 9 categories: Metabolism, Storage, Photosynthesis, Amino Acid, Allergy, Stress, Protein Synthesis, Enzyme and, Hypothetical protein. The gluten includes two different components, high molecular weight glutenin subunits and low molecular weight glutenin subunits and gliadins. Some selected protein spots were detected to be (i) gluten, which is responsible for roughness and viscoelasticity for bread making quality (ii) stress proteins (biotic and abiotic) associated with salt, cold, heat tolerance, disease (iii) pathogen related proteins, and (iv) allergenic proteins responsible for allergy in humans, (v) puroindoline- a & b (encoding PinA and PinB gene)that is responsible for grain texture related to baking performance and roughness and other molecular functions such as antibiotic / toxin / antimicrobial activities, that contribute to the defense mechanism of the plant against predators. Moreover, to gain a better understanding of proteome analysis and identify the pre-harvest sprouting responsible proteins, we carried out a comparative proteomic analysis in pre-harvest sprouting wheat seeds between susceptible and resistant cultivars.
Keywords: Wheat; pre-harvest sprouting; susceptible; resistance; proteomics analysis; mass spectrometry.
Pages 110-119 Full Text PDF
Antioxidant potentials protect Vigna radiata (L.) Wilczek plants from soil cobalt stress and improve growth and pigment composition
C. Abdul Jaleel, K. Jayakumar, Zhao Chang-Xing, M.M. Azooz,
The experiments were conducted in earthen pots lined with polythene sheet to find out the effect of different concentrations of cobalt (0, 50, 100, 150, 200 and 250 mg/kg soil) on various morphological parameters, photosynthetic pigment contents and antioxidant enzyme activities on greengram (Vigna radiata (L.) Wilczek). Plants were watered to field capacity daily. Plants were thinned to a maximum of five per pot. The data for various morphological parameters such as, root and shoot length, number of nodules, dry weight of root and shoot and photosynthetic pigments such as chlorophyll ‘a’, chlorophyll ‘b’, total chlorophyll and carotenoids content were collected on 30 days after sowing (DAS). Antioxidant enzymes like catalase, peroxidase and polyphenol oxidase activities were analysed from both control and treated plants. All the growth parameters and pigment contents increased at 50 mg/kg cobalt level in the soil, when compared with control. Further increase in cobalt level (100-250 mg/kg) in the soil had a negative impact upon all studied parameters. From these results it is clear that Antioxidant potentials acts as a protective mechanism in Vigna radiata under soil cobalt stress.
Keywords: antioxidant potentials; cobalt; morphological parameters; photosynthetic pigment; greengram; Vigna radiata
Pages 120-126 Full Text PDF
Study of genetic variability of Wormwood capillary (Artemisia capillaris) using inter simple sequence repeat (ISSR) in Pahang region, Malaysia.
Mohammad Shafie B. Shafie, Sayed M. Zain Hasan and Ramisah M. Shah
The genetic variability of five individuals of Artemisia capillaris from state of Pahang, Malaysia was examined using Inter Simple Sequence Repeat (ISSR) technique. Samples were collected from different regions in Pahang State. Results showed distinctive ISSR banding pattern in this species. Twenty five primers were applied and only ten were selected (807, 809, 825, 834, 841, 862, 866, 876, nIssr 1 and nIssr 3) as reliable amplifying ISSR markers. A total of 90 ISSR bands including 62 polymorphic (68.89%) with amplicon size ranging from 150-2500 bp were scored. Genetic distance for samples ranged from 0.0500 to 0.4200. For similarity index samples were ranged from 0.6400 to 0.9545.
Keywords: Artemisia capillaries; phylogenic; variability; polymorphism; wormwood.
Pages 127-134 Full Text PDF
May 2009 issue
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