scholarly journals Establishment and Characterization of Callus and Cell Suspension Cultures of Selected Sorghum bicolor (L.) Moench Varieties: A Resource for Gene Discovery in Plant Stress Biology

Agronomy ◽  
2019 ◽  
Vol 9 (5) ◽  
pp. 218 ◽  
Author(s):  
Elelwani Ramulifho ◽  
Tatenda Goche ◽  
Johann Van As ◽  
Toi John Tsilo ◽  
Stephen Chivasa ◽  
...  

Sorghum, a naturally drought tolerant crop, is genetically diverse and provides a wide gene pool for exploitation in crop breeding. In this study, we experimentally assessed friable callus induction rates of seven sorghum varieties using shoot explant for the generation of cell suspension cultures. The cell suspensions were characterized in terms of cell growth and viability profiles as well as gene expression following 400 mM sorbitol-induced osmotic stress for 72 h. Only ICSB 338, a drought susceptible variety, was readily amenable to friable callus formation. Cell culture growth plots of both ICSB 338 and White sorghum (used as a reference line) depicted typical sigmoidal curves. Interestingly, Evans blue assay showed that ICSB 338 cell cultures are more susceptible to osmotic stress than the White sorghum cells. The osmotic stress treatment also triggered differential expression of eight target genes between the two cell culture lines. Overall, these results suggest that the genetic diversity of sorghum germplasm influences friable callus induction rates and molecular responses to osmotic stress, and could be further exploited in plant stress biology studies. Therefore, we have developed a valuable resource for use in molecular studies of sorghum in response to a range of biotic and abiotic stresses.

Author(s):  
P.T.M. Tram ◽  
N.K. Suong ◽  
L.T.T. Tien

Background: Belonging to the Boraginacae family, Ehretia asperula Zoll. et Mor., called “Xa den”, is a precious medicinal plant also known as the “cancer tree” by the Muong ethnic group in Hoa Binh Province, Vietnam. Xa den has been demonstrated to inhibit the development of malignant tumors, reduce oxidation and enhance the human immune system. This research focused on examining friable callus induction from young stems of Ehretia asperula Zoll. et Mor. Methods: Samples of Xa den were less than two years old. Young stems with 2 to 6 leaves served as explants for callus induction. Explants placed on autoclaved B5 nutrients incubated at 25oC, in the dark. The testing factors were concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) and Benzyl adenine (BA), types and concentrations of sugars.Result: Friable callus was induced on B5 medium with 0.4 mg/L of 2,4-D, 0.1 mg/L of BA and 30 g/L of glucose at the highest rate (100%). Additionally, callus grew best after 5 weeks of culture weighing 0.194 g. Friable callus was used as material for cell suspension cultures. After two weeks, the Xa den cell suspension cultures contained single cells and small cell clumps. The liquid medium had changed from dark yellow to light brown.


2006 ◽  
Vol 61 (3-4) ◽  
pp. 251-256 ◽  
Author(s):  
Hua Cheng ◽  
Long-Jiang Yu ◽  
Qiong-Yue Hu ◽  
Shan-Cai Chen ◽  
You-Ping Sun

An efficient procedure has been developed for callus induction and cell suspension cultures of C. saxicola for the first time. Explant selection was carried out among leaf, stem and root to select a suitable type of explants capable of higher callus formation. Leaf explants thus selected showed maximum response to callus induction (67.1%). Modified B5 medium supplemented with 0.5 mg l−1 2,4-D plus 2 mg l−1 BA was the most favorable medium for callus formation with the highest induction rate (94.8%) and greatest fresh weight of callus (1.7 g per explant). Cell suspension cultures were established by transferring 2-8 g fresh callus to 80 ml liquid B5 medium. An inoculum size of 8 g produced the greatest biomass accumulation, dehydrocavidine and berberine productions, which was 13.1 g l−1, 8.0 mg l−1 and 4.1 mg l−1, respectively. In response to various sucrose concentrations from 10 g l−1 to 80 g l−1, cultures with 60 g sucrose l−1 not only produced the highest dry biomass (18.5 g l−1) but also the highest formation of dehydrocavidine (11.6 mg l−1) and berberine (7.6 mg l−1). These prepared cell suspension cultures provided a useful material for further regulation of alkaloid biosynthesis and for enhanced production of valuable alkaloids on a large scale.


2017 ◽  
Vol 6 (2) ◽  
pp. 38 ◽  
Author(s):  
Liyan Jin ◽  
Yang Yang ◽  
Wenjie Gao ◽  
Mingxue Gong ◽  
Jijia Wang ◽  
...  

Dendranthema indicum var. aromaticum is an important aroma plant in genus Dendrathema, and the establishment of callus cultures and cell suspension cultures is the basement of further protoplast fusion studies, which make it possible to breed new fragrant chrysanthemum. In this study, the effects of different plant growth regulating substances in different concentrations on callus induction were investigated with stem segments, leaves, petioles as explants. The results showed that the optimal explants were lower stem segments according to the percentage of callus formation, callus hardness, growth potential and shoot differentiation. The optimal induction mediums were MS supplemented with 1.0 mg.l-12.4 D and 0.2 mg.l-1 6-BA. The cell suspension culture system was established by using the subculture calli. The results showed that the suitable inoculum size was 2g and the suitable cell suspension culture medium was MS supplemented with 0.2 mg.l-1 6-BA and 0.5 mg.l-1 2,4-D. The time course of cell growth showed that the greatest cell fresh weight appeared on day 14 and the highest cell viability on day 3.


2017 ◽  
Vol 16 (1) ◽  
pp. 1-11
Author(s):  
Aryani Leksonowati ◽  
Witjaksono Witjaksono ◽  
Diah Ratnadewi

Aquilaria malaccensis Lam. is a plant species producing fragrant woody material that contains some resin. The compounds can be used as medicine and perfume. Sesquiterpenoid, one group of compounds has been found being synthesized and subsequently extracted from callus and cell suspension culture of Aquilaria species. The aim of this research was to find a method of producing friable calli and cell suspension cultures from leaves or internodes of A. malaccensis in vitro by using suitable plant growth regulators; cell suspension that will suitably serve as material to produce sesquiterpenoid afterwards. Calli were established in almost all treatments of auxin-cytokinin on both leaves and internod explants. The treatment of 10 mg/L IBA induced the highest percentage of callus coverage from leaves with a rather compact structure. The combined treatment of 1–2 mg/L 2.4-D and 0.2–0.3 mg/L BA induced friable callus formation in more than 80% of cultures with 27–32% callus coverage percentage.  The use of 2,4-D induced a better formation of cell suspension than Picloram, with maximum volume up to 7 mL. Cell suspension culture with fine and homogenous aggregate could be established in the medium supplemented with 0.5 –1 mg/L 2,4-D.


1982 ◽  
Vol 37 (10) ◽  
pp. 857-860 ◽  
Author(s):  
Joachim Stöckigt ◽  
Karl-Heinz Pawelka ◽  
Ana Rother ◽  
Brigitte Deus

Abstract Cell suspension cultures of Stemmadenia tomentosa synthesized under normal growth condition the eight major indole alkaloids: (-)-tabersonine, (-)-minovincinine, (+)-conoflorine (voaphyl-line), condylocarpine, (+)-tubotaiwine (dihydrocondylocarpine), (-)-norfluorocurarine (vin-canine), (-)-vinervine, and (-)-coronaridine. These alkaloids consist of the three different types, Aspidosperma, Strychnos and Iboga. In contrast, cultures o f Voacanga africana produced mainly one alkaloid group (Aspidosperma-type) represented by (-)-tabersonine, lochnericine and (-)-minovincinine. Therefore this cell culture seems to be qualified for investigation concerning the biosynthesis of Aspidosperma alkaloids.


1988 ◽  
Vol 43 (7-8) ◽  
pp. 536-544 ◽  
Author(s):  
Susanne Daniel ◽  
Walter Hinderer ◽  
Wolfgang Barz

The extractable activities of thirteen enzymes of primary and secondary metabolism have been measured in chickpea (Cicer arietinum L.) cell suspension cultures after treatment with an elicitor from the fungus Ascochyta rabiei (Pass.) Lab. The cell culture, derived from the A. rabiei resistant cultivar ILC 3279, constitutively accumulated the isoflavones biochanin A and formononetin together with their 7-O-glucosides and the 7-O-glucoside-6″-malonates. After elicitor application the cells rapidly form the pterocarpan phytoalexins medicarpin and maackiain. Among the enzymes of primary metabolism only the glucose 6-phosphate dehydrogenase exhibited a significant increase in activity with a maximum four hours after application of the elicitor. In phenylpropane metabolism the activities of phenylalanine ammonia lyase and chalcone synthase were enhanced by the elicitor and exhibited highest levels after four hours. In contrast the chalcone isomerase activity was not influenced by the elicitor. A substantial enhancement occurred with the isoflavone 7-O-glucosyltransferase activity eight hours after elicitor application. The results suggest that in this cell culture the elicitor-induced biosynthesis of pterocarpan phytoalexins was accompanied with a rapid and transient increase of those enzyme activities which are located at branching points of related pathways, i.e. pentose phosphate cycle, general phenylpropane metabolism, flavonoid formation and isoflavone conjugation.


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