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1

The effect of polyamines on the efficiency of multiplication and rooting of Withania somnifera (L.) Dunal and content of some withanolides in obtained plants

100%
Sivanandhan G., Mariashibu T. S., Arun M., Rajesh M., Kasthurirengan S., Selvaraj N., Ganapathi A.
Acta Physiologiae Plantarum
|
2011
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tom 33
|
nr 6
An efficient mass multiplication protocol was developed for Withania somnifera (L.) Dunal from nodal explants of field-grown plants on Murashige and Skoog medium (MS) supplemented with 6-benzyladenine (BA) [1.5 mg L⁻¹], indole-3-acetic acid (IAA) [0.3 mg L⁻¹] and with the addition of polyamine, spermidine (20 mg L⁻¹) (shoot multiplication medium). A total of 46.4 shoots were obtained from nodal explants and they were elongated in the same medium in a culture duration of 6 weeks. The elongated shoots produced roots in MS medium fortified with putrescine (20 mg L⁻¹) after 4 weeks, and all the rooted plants were successfully hardened and acclimatized with a survival rate of 100%. An average of 276 shoots (46 × 6) was produced when at least six nodal explants obtained from each of the 46 in vitro grown shoots were cultured by microcutting method in the same shoot multiplication medium. On an average, 12,696 plants could be produced from all the shoots (276 × 46) by microcuttings in a period of 7 months. HPLC revealed a significant increase in the quantities of withanolide A, withanolide B, withaferin A and withanone in the leaves, stems, and roots of in vitro regenerated plants compared to the field-grown parent plants. Ploidy analysis using flow cytometry revealed genetic stability of in vitro regenerated plants. This protocol will be useful for scale-up production of withanolides on commercial scale.
2

Factors influencing podophyllotoxin production in adventitious root culture of Podophyllum hexandrum Royle

84%
Rajesh M., Sivanandhan G., Arun M., Vasudevan V., Theboral J., Girija S., Manickavasagam M., Selvaraj N., Ganapathi A.
Acta Physiologiae Plantarum
|
2014
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tom 36
|
nr 04
To determine the possibility of generating the Podophyllotoxin accumulation using Podophyllum hexandrum adventitious roots derived from root segments, several nutrient constituents (carbon sources, media strength, initial medium pH, ammonium and nitrate proportion and phosphate ratio) were evaluated in culture. The maximum biomass accumulation was recorded in 0.50 MS medium containing 3 mg/l indole-3-butyric acid and 2 % sucrose, and the maximum accumulation of Podophyllotoxin was documented in the same strength of MS medium with 6 % of sucrose. When the initial medium pH was on 6 in the optimized MS medium, the biomass and Podophyllotoxin accumulations were highest. The lower concentration of ammonium (10 mM) in combination with a moderate concentration of nitrate (20 mM) was found ideal for maximum accumulations of biomass and Podophyllotoxin. Maximum Podophyllotoxin accumulation (6.4 mg/g dry weight) was recorded at the higher concentration of phosphate (2.25 mM), and lower concentration of phosphate (1.25 mM) showed highest growth accumulation. The outcome of the present work will be helpful for the large-scale cultivation of adventitious root for the production of Podophyllotoxin.
3

Vacuum infiltration enhances the Agrobacterium-mediated genetic transformation in Indian soybean cultivars

84%
Mariashibu T. S., Subramanyam K., Arun M., Mayavan S., Rajesh M., Theboral J., Manickavasagam M., Ganapathi A.
Acta Physiologiae Plantarum
|
2013
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tom 35
|
nr 01
For the first time we have developed a reliable and efficient vacuum infiltration-assisted Agrobacteriummediated genetic transformation (VIAAT) protocol for Indian soybean cultivars and recovered fertile transgenic soybean plants through somatic embryogenesis. Immature cotyledons were used as an explant and three Agrobacterium tumefaciens strains (EHA 101, EHA 105, and KYRT 1) harbouring the binary vector pCAMBIA1301 were experimented in the co-cultivation. The immature cotyledons were pre-cultured in liquid somatic embryo induction medium prior to vacuum infiltration with the Agrobacterium suspension and co-cultivated for 3 days on co-cultivation medium containing 50 mg l-1 citric acid, 100 lM acetosyringone, and 100 mg l-1 L-cysteine. The transformed somatic embryos were selected in liquid somatic embryo induction medium containing 10 mg l-1 hygromycin and the embryos were germinated in basal medium containing 20 mg l-1 hygromycin. The presence and integration of the hpt II and gus genes into the soybean genome were confirmed by GUS histochemical assay, polymerase chain reaction, and Southern hybridization. Among the different combinations tested, high transformation efficiency (9.45 %) was achieved when immature cotyledons of cv. Pusa 16 were pre-cultured for 18 h and vacuum infiltrated with Agrobacterium tumefaciens KYRT 1 for 2 min at 750 mm of Hg. Among six Indian soybean cultivars tested, Pusa 16 showed highest transformation efficiency of 9.45 %. The transformation efficiency of this method (VIAAT) was higher than previously reported sonication-assisted Agrobacterium-mediated transformation. These results suggest that an efficient Agrobacteriummediated transformation protocol for stable integration of foreign genes into soybean has been developed.
4

Effect of culture conditions, cytokinins, methyl jasmonate and salicylic acid on the biomass accumulation and production of withanolides in multiple shoot culture of Withania somnifera (L.) Dunal using liquid culture

84%
Sivanandhan G., Rajesh M., Arun M., Jeyaraj M., Dev G. K., Arjunan A., Manickavasagam M., Muthuselvam M., Selvaraj N., Ganapathi A.
Acta Physiologiae Plantarum
|
2013
|
tom 35
|
nr 03
The influence of cytokinins and culture conditions including medium volume, harvest time and elicitation with abiotic elicitors (SA/MeJ) have been studied for the optimal production of biomass and withanolides in the multiple shoot culture of Withania somnifera. Elicitation of shoot inoculum mass (2 g 1-1 FW) with SA at 100 μM in the presence of 0.6 mg 1-1 BA and 20 mg 1-1 spermidine for 4 h exposure time at the 4th week in 20 m1 liquid medium recorded higher withanolides production (withanolides A [8.48 mg g-1 DW], withanolides B [15.47 mg g-1 DW], withaferin A [29.55 mg g-1 DW] and withanone [23.44 mg g-1 DW]), which were 1.14 to 1.18- fold higher than elicitation with MeJ at 100 μM after 5 weeks of culture. SA-elicited cultures did not exhibit much variation in biomass accumulation when compared to control. This cytokinin induces and SA-elicited multiple shoot culture protocol provides a potential alternative for the optimal production of biomass and withanolides utilizing liquid culture.
5

High-efficient Agrobacterium-mediated in planta transformation in black gram (Vigna mungo (L.) Hepper)

68%
Kapildev G., Chnnathambi A., Sivanandhan G., Rajesh M., Vasudevan V., Mayavan V., Arun M., Jeyaraj M., Alharbi S. A., Selvaraj N., Ganapathi A.
Acta Physiologiae Plantarum
|
2016
|
tom 38
|
nr 08
6

Estimation of cytogenetic risk among coke oven workers exposed to polycyclic aromatic hydrocarbons

68%
Sureshkumar S., Balachandar V., Devi S. M., Arun M., Karthickkumar A., Balamuralikrishnan B., Sankar K., Mustaqahamed S. A. K., Dharwadkar S. N., Sasikala K., Cho S.-G.
Acta Biochimica Polonica
|
2013
|
tom 60
|
nr 3
Polycyclic aromatic hydrocarbons (PAHs) result from the incomplete combustion of natural or synthetic organic materials. The working environment at a coke plant can negatively affect the employed workers who were exposed to coke oven emissions containing PAHs, which formed and released into the environment by the process of pyrolysis of coke. This study aims to analyze the relationship between the exposure of PAHs and the risk of genetic damages such as chromosomal alteration (CA), micronucleus (MN), and DNA damage (PCR-RFLP) in peripheral blood lymphocytes of 27 coke oven workers and equal number of control subjects. The exposed subjects and controls were divided into two groups based on their age (group I<35 years and group II ≥35 years). The exposed subjects were further classified into two groups based on the exposure period (<12 years and ≥12 years). The frequencies of CA and MN in exposed subjects are relatively high with respect to controls. The XRCC1 399 Arg/gln polymorphism showed a substantial smaller difference in allele frequencies between exposed and control subjects. Based on present data, it was concluded that coke oven workers under risk should be monitored for adverse effects of the any long-term exposure.
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