02795nas a2200217   4500008004100000245009300041210006900134260000800203300001200211490001000223520206100233100002302294700002002317700002302337700002102360700001902381700001602400700001802416700002102434856012202455       2017                        eng d00aPyrethrin accumulation in elicited hairy root cultures of chrysanthemum cinerariaefolium0 aPyrethrin accumulation in elicited hairy root cultures of chrysa  cAPR  a365-3760 v 81  3 aThe flowers of Pyrethrum (Chrysanthemum cinerariaefolium) are known to contain Pyrethrins that are naturally occurring potential insecticide. Hairy roots were induced from leaves of C. cinerariaefolium using Agrobacterium rhizogenes strain A4. The root clones were characterized in to four groups i.e. thick, unbranched (D2 and D5), thin, highly branched (D3), thick, branched (B2) and thick, highly branched (D1, D6). Six established hairy root clones showed the presence of pyrethrin and were selected for elicitation studies. Growth kinetics studies revealed highest growth index in hairy root clone D1 (592.0) followed by D6 and D3 on dry weight basis after 40 days of culture. The maximum pyrethrin content was found in the clone D3 (7.2 mg/g dw) which is comparable to the flowers obtained from the variety "Avadh". Hairy root clone D2 (5.2 mg/g dw) and D6 (1.3 mg/g dw) contained pyrethrin but in less amount as compared to clone D3. The PCR analysis showed the presence of rol B and rol C genes in all the six hairy root clones while rol A was detected only in D2 clone. The methanolic extract of D3 clone showed antifungal activities against phytopathogenic fungal strains which were found maximum against Curvuleria andropogonis followed by Colletotrichum acutatum and Rhizoctonia solani. Hairy root clones D2, D3 and D6 were elicited with culture filtrate of endophytic fungus (Fusarium oxysporum) and bacteria (Bacillus subtilis). The culture filtrate (4.0 %v/v) of both the fungal and bacterial origin was found to be effective in enhancing the pyrethrin content in all the tested hairy root clones. Clone D3 showed maximum pyrethrin content on elicitation with F. oxysporum (9.7 mg/g dw) and B. subtilis (9.7 mg/g dw) culture filtrate, which is 32 % higher than the non elicited D3 hairy roots (7.2 mg/g dw). F. oxysporum also enhanced the hairy root growth resulting into the higher biomass yield of D3 (50 %) and D2 (76 %) in comparison to control non elicited hairy root clones of D3 and D2, respectively leading to higher pyrethrin yield.1 aKhan, Shamshad, A.1 aVerma, Priyanka1 aBanerjee, Suchitra1 aChaterjee, Arnab1 aTandon, Sudeep1 aKalra, Alok1 aKhaliq, Abdul1 aRahman, Laiq, Ur  uhttps://library.ncl.res.in/content/pyrethrin-accumulation-elicited-hairy-root-cultures-chrysanthemum-cinerariaefolium03264nas a2200277   4500008004100000022001400041245017200055210006900227260000800296300001200304490000800316520228800324653002402612653001402636653002602650653002602676653000902702653001502711653002202726100002302748700002002771700001902791700002002810700002802830856012802858       2018                        eng d  a0926-669000aDevelopment of enhanced hypericin yielding transgenic plants and somaclones: high throughput direct organogenesis from leaf and callus explants of Hypericum perforatum0 aDevelopment of enhanced hypericin yielding transgenic plants and  cJAN  a544-5540 v1113 a<p>Hypericum perforatum is well known for its anti-depressent properties due to the presence of hypericins and hyperforins. The direct regeneration protocol from leaf explants and Agrobacterium tumefaciens mediated genetic transformation is a prerequisite for gene transfer studies in this valuable plant system as H. perforatum has shown its recalcitrance to these studies previously. The aim of generating transgenic shoots or somaclones in the present study is to select high hypericine yielding H. perforatum plants. High throughput direct bud organogenesis from leaf explants has been achieved for the first time in this plant system on media combination of MS + 5.0 mg/l BAP + 1.0 mg/l. This medium interestingly also worked as the indirect regeneration media for generation of somaclones from callus cultures. Thus far optimized single media can be used for direct and indirect regeneration in H. perforatum. The direct regeneration frequency of plantlets from leaf explants was found to be 80% while 100% was the regeneration frequency noted for indirect plantlet regeneration from callus cultures. The optimized direct regeneration protocol from leaf explants was utilized for A. tumefaciens (harbouring pCAMBIA 1301) mediated genetic transformation studies with the aim of establishing transgenic lines of H. perforatum. The resultant study successfully gave rise to kanamycin resistant GUS positive shoots with transformation efficiency of 3136 +/- 3.6. A total of four transgenic H. perforatum clones namely T2, T3, T5 and T8 with different growth parameters were established, where highest of fivefold increase in hypericin content was recorded by T5 (276.8 +/- 9.2 mu g/g dry wt.) in comparison to control non transformed plants (35.6 +/- 2.7 mu g/g dry wt.). Interestingly the presence of serotonin and melatonin was also detected in transgenic plants on TLC basis. The transgenic nature of the plants was confirmed by PCR with amplification of uidA gene. Apart from it, from indirect regeneration from callus, total of ten somaclones were generated. The ISSR profiling of somaclones scored a total of 156 bands, among which 118 were polymorphic in nature. The glasshouse established soma clones showed up to twofold increase in hypericin content on HPLC analysis.</p>10aDirect regeneration10aHypericin10aHypericum perforation10aIndirect regeneration10aISSR10aSomaclones10aTransgenic plants1 aKhan, Shamshad, A.1 aVerma, Priyanka1 aArbat, Akshata1 aGaikwad, Sushma1 aParasharami, Varsha, A.  uhttps://library.ncl.res.in/content/development-enhanced-hypericin-yielding-transgenic-plants-and-somaclones-high-throughput