In Vitro Selection in Wheat to Recover Mutants which Overproduce Lysine and Threonine

Crop Scienc

In vitro mass propagation of Typhonium flagelliforme as affected by plant growth regulators

Tubers were used as explants in in vitro mass propagation of Rodent Tuber (Typhonium flagelliforme). The explants were obtained from sterile plantlets and placed in shoot induction medium containing basal salts of Murashige and Skoog (MS) and various concentrations of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA). Treatment containing 5 mg/l (w/v) of BAP with 1 mg/l (w/v) of NAA produced the highest number of shoots per explant (29.17) after 12 weeks of culture and also the highest mean fresh weight of shoots formed in treatment containing 5 mg/l (w/v) of BAP with 1 mg/l (w/v) of NAA. For ex vitroestablishment, well- rooted plantlets were transferred in potting medium containing peatmoss, perlite and vermiculite (3:1:1)

Genetic diversity evaluation of Cumin (Cumin cyminum L.) based on phenotypic characteristics.

In order to assess the genetic diversity of cumin and determine the traits effective on seed yield and cumin- aldehyde, forty nine cumin ecotypes which they are sub-populations belonged to nine populations from different provinces of Iran were evaluated based on morphological and biochemical traits. Results indicated a significant variation for all the measured traits among and within populations derived from different provinces. Kerman and Esfahan populations showed the best performance based on the phenotypic data, while Yazd had almost the lowest levels of traits. Correlation analysis showed number of seed per umbel and umbel per plant had highest relationship with seed yield. Path analysis also demonstrated that number of umbel per plant and number of seed per umbel had the most direct effects on seed yield and were identified as the most effective factors on seed yield. Cumin aldehyde was mostly correlated by number of umbel per plant. The present study showed that different qualitative characteristics such as seeds with light color and without trichome and leaves without trichome, alternate and large pods of Petiole tend to produce high seed yield. Pattern analysis of different populations based on first two main principal components categorized the measured genotypes in to three groups: Pars, Northern_Khorasan, Golestan, Semnan and Yazd (Group1), Southern_Khorasan and Khorasan_Razavi (Group2) Kerman and Esfahan (Group3), which the third group are high yielding genotypes with different genetic background can be advised for cultivation and breeding programs. So the available genetic diversity among the Iranian cumin populations can be lead to produce high yielding population of cumin

Callus induction in pineapple (Ananas comosus L.) cv. Moris and Josapine.

The induction of callus from Meristemic Globular Bodies (MGB) of two pineapple cultivars, namely Moris and Josapine, under six concentration levels of auxin NAA and six concentration levels of 2,4-D in Murashige and Skoog solid media, was investigated. 2,4-D auxin treatments failed to induce calli in both cultivars. However, 53.71, 75.19 and 85.93 μM levels of auxin NAA caused calli induction in Moris while levels 32.22, 53.71 and 75.19 μM also induced calli Josapine. The percentage of MGB calli formation increased with increasing time of culturing. At 6 weeks of culturing, 83% of Moris MGB explants formed calli on 85.93 μM NAA, while 50% of Josapine MGB explants formed calli on 53.71 μM NAA. Calli cultures have been an essential tool in the in vitro selection of desirable plants under manipulated conditions and from in vitro mutations via somaclonal variation. More importantly, calli are increasingly used for the application of cellular level genetic modification techniques such as the Agrobacterium-mediated transformation, particle bombardment and protoplast isolation and fusion. In this study, auxin NAA successfully initiated and proliferated calli in Moris and Josapine globular meristemic cultures

Factors influencing in vitro tuberization of Chlorophytum borivilianum in solid culture

The present study describes an efficient technique of microtuberization of Chlorophytum borivilianum. In this study, young shoot buds of C. borivilianum were cultured on solid MS medium supplemented with 30, 60 and 90 gl-1 sucrose either individually or in combination with 0, 315, 630, 950, 1265 and 1580 μM 2-chloroethyl-trimethylammonium chloride (CCC). The highest mean number of microtubers was significantly enhanced in the presence of CCC at 950, 1265 and 1580 μM after 8 weeks of culture whereas the lowest occurred in the absence of CCC. Increasing the sucrose only stimulated microtuber elongation. The number of microtubers produced was strongly enhanced by the concentration of up to 60 gl-1 sucrose in the medium compared with 30gl-1 sucrose, but at higher level of 90 gl-1 sucrose, microtuber production declined. In the presence of sucrose, number of microtuber increased with increasing CCC level up to 1265 μM particularly at 60gl-1 with CCC mean number of microtuber per explants was more pronounced (3.5). Meanwhile, using 90gl-1 sucrose with or without CCC showed higher microtuber length which indicates that at high sucrose concentration, CCC has no significant role on tuber elongation. Results showed the most suitable combination for in vitro tuberization of C. borivilianum was 950 μM CCC with 60 gl-1 sucrose

Assessment of antioxidant and cytotoxicity activities of saponin and crude extracts of Chlorophytum birivilianum

The present paper focused on antioxidant and cytotoxicity assessment of crude and total saponin fraction of Chlorophytum borivilianum as an important medicinal plant. In this study, three different antioxidant activities (2,2-diphenyl-1-picrylhydrazyl radical scavenging (DPPH), ferrous ion chelating (FIC), and β-carotene bleaching (BCB) activity) of crude extract and total saponin fraction of C. borivilianum tubers were performed. Crude extract was found to possess higher free radical scavenging activity (ascorbic acid equivalents 2578 ± 111 mg AA/100 g) and bleaching activity (IC50 = 0.7 mg mL−1), while total saponin fraction displayed higher ferrous ion chelating (EC50 = 1 mg mL−1). Cytotoxicity evaluation of crude extract and total saponin fraction against MCF-7, PC3, and HCT-116 cancer cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) cell viability assay indicated a higher cytotoxicity activity of the crude extract than the total saponin fraction on all cell lines, being most effective and selective on MCF-7 human breast cancer cell line

In vitro mutagenesis of Etlingera elatior (Jack) and early detection of mutation using RAPD markers.

Mutation breeding techniques in combination with tissue culture and molecular marker methods provide a powerful tool for improvement of vegetatively propagated plants. The aim of this study was to develop a protocol for shoot regeneration and mutation induction of Etlingera elatior. The results of irradiation on in vitro buds of E. elatior showed LD50 to be 10 Gy, with the survival of explants being sharply reduced at this dosage. All 8 selected gamma irradiated regenerants were differentiated from the untreated control based on the banding patterns obtained using 9 primers, which generated 59 reproducible bands, whereby 35 (55.31%) were found to be polymorphic. Jaccard’s coefficient of similarity values ranging from 0.537 to 0.860 were indicative of the level of genetic variation among the mutants studied. For comparison between the potential lines (PL) and the control, a maximum similarity value(0.814) was observed in PL1 mutant, while the minimum value (0.537) was observed in PL7. In summary, a combination of irradiation, regeneration, multiplication, and random amplification of polymorphic DNA (RAPD) analysis for early screening of mutants can speed up the breeding program of E. elatior

Plant regeneration of Brassica oleracea sub sp. italica (Broccoli) CV Green Marvel as affected by plant growth regulators

Hypocotyls and shoot tips were used as explants in in vitro plant regeneration of broccoli (Brassica oleracea subsp. italica) cv. Green Marvel. The explants were excised from sterile germinated seedlings and placed on shoot induction medium containing basal salts of Murashige and Skoog (MS) and various concentrations of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA). The highest percentage of hypocotyl explant producing shoot (96.67%) and the highest mean number of shoots produced per hypocotyl explant (6.03) were obtained on 3 mgL-1BAP. Meanwhile, the highest percentage of shoot tip explant producing shoot (100%) and highest number of shoot produced per shoot tip explant (3.76) were recorded on 5 mgL-1 BAP. For rooting of shoots, NAA, indoleacetic acid (IAA) and indolebutyric acid (IBA) at 0, 0.2 and 1 mgL-1 were applied. Highest percentage of shoots with roots (100%) and highest mean number of roots produced per shoot (6.5) occurred on medium with 0.2 mgL-1 IBA, while the maximum root length (2.46 cm) was attained on MS medium without plant growth regulator (MSO). Plantlets were successfully acclimatized in potting medium containing peatmoss, perlite, and vermiculite (3:1:1)

Incidence of fern contamination in nodal segment cultures of Shorea parvifolia dyer

The greatest drawback in large scale micropropagation of tropical woody forest species is high contamination of cultures. In developing a sterilization protocol for micropropagation of Shorea parvifolia Dyer utilizing nodal segments excised from nursery-grown seedlings, it was found that washing 20% (v/v) with Clorox solution for 18 minutes was the best. After six weeks of culture in WPM media supplemented with 10-5 M BAP (apart from fungal and bacterial contamination), the nodal segments developed hair-like structures which were amenable to subculture. Upon subculture, green leafy structures developed from the mass of hairy structure after six weeks. These later developed into ferns which are normally found as epiphytes on older forest trees, known as Asplenium nidus

Genetic stability of in vitro multiplied Phalaenopsis gigantea protocorm-like bodies as affected by chitosan

Chitosan is a carbohydrate polymer derivative of chitin which presents in shell of crustaceans. This biopolymer is a non toxic and environmentally friendly, considered as a plant growth stimulator in some plant species. The present study investigates the effects of chitosan and media types on multiplication and genetic stability of Phalaenopsis gigantea protocorm-like bodies (PLBs). PLBs were inoculated in liquid New Dogashima Medium (NDM) and Vacin and Went (VW) supplemented with various concentrations of chitosan (0, 5, 10, 15, 20 and 25 mg/L). The highest PLB multiplication was observed on VW and NDM supplemented with 10 mg/L chitosan with mean number of PLBs 177 and 147, respectively. Chitosan promoted the formation of juvenile leaves and the highest number was observed in NDM supplemented with 20 mg/L chitosan with mean number of 66 leaves after 8 weeks of culture. Genetic stability was assessed among mother plant and secondary PLBs after 2, 4, 6, and 8 weeks of culture in liquid media. 8 out of 10 ISSR markers produced a total of 275 clear and reproducible bands with mean of 6.9 bands per primer. The secondary PLBs produced during sub-culturing process of chitosan treated liquid culture were genetically uniform and similar to mother plant