02920nas a2200253   4500008004100000022001400041245010000055210006900155260000800224300001200232520206200244653003102306653002002337653001602357653002802373653001102401653001402412653002502426653001602451100002602467700002002493700002802513856012502541       2022                        eng d  a0167-685700aHomo and heterologous expression of the HpPKS2 gene in Hypericum perforatum and Bacopa monnieri0 aHomo and heterologous expression of the HpPKS2 gene in Hypericum  cJAN  a215-2153 a<p>Hypericum perforatum has been known to produce hypericin and hyperforin that are used in treating mild to moderate depression. The HpPKS2 (H. perforatum polyketide synthase 2) gene is assumed to play a role in hypericin biosynthesis. The HpPKS2 gene was overexpressed in homologous H. perforatum in vitro grown plants through Agrobacterium tumefaciens-mediated genetic transformations. It leads to the establishment of seven glass house acclimatized transgenic lines. Among them, the HP12 transgenic plant showed 9.8 fold enhancement in hypericin content (379.4 +/- 10.3 mu g/g dry wt) followed by 3-fold in HP41 (117.1 +/- 4.5 mu g/g dry wt) as compared to control plants. This was further supported by the real-time PCR studies where it registered up to 5 fold enhancement of HpPKS2 gene expression. On the other hand, the heterologous expression of the HpPKS2 gene in Bacopa monnieri resulted in the establishment of five transgenic plant clones that were successfully acclimatized under glasshouse conditions. Among them, BT4 was found to be very slow-growing. The BT3 line showed maximum expression of the HpPKS2 gene which surprisingly also upregulates the expression of the other metabolic pathway genes of B. monierri namely isopentyl- diphosphate delta isomerase (IDDI), squaline synthase (SQS) and acetyl CoA C acetyltransferase (AA). The HPLC analysis in the heterologous system revealed the maximum production of bacopaside I (9.86 +/- 1.0 mg/g dry wt), bacopaside II (5.89 +/- 0.9 mg/g dry wt) and bacopasaponin C (3.59 +/- 0.3 mg/g dry wt) by transgenic lines BT8, BT3 and BT4, respectively. This enhancement in bacopaside I, bacopaside II and bacopasaponin C production was more than 26-fold, 5-fold and 21-fold, respectively in comparison to the control non transformed plants. Key message Overexpression of the HpPKS2 gene in H. perforatum led to higher hypericin content in the native system while its expression in the heterologous system i.e. B. monnieri also improves bacopaside I, bacopaside II and bacopasaponin C production.</p>
10aAgrobacterium tumefacience10aBacopa monnieri10aBacopasides10aHeterologous expression10aHpPKS210aHypericin10aHypericum perforatum10aTransgenics1 aKhan, Shamshad, Ahmad1 aVerma, Priyanka1 aParasharami, Varsha, A.  uhttps://library.ncl.res.in/content/homo-and-heterologous-expression-hppks2-gene-hypericum-perforatum-and-bacopa-monnieri02949nas a2200385   4500008004100000022001400041245017500055210006900230260000800299300001200307490000800319520168600327653001902013653001802032653001502050653001402065653001402079653002202093653001402115653000902129653001202138653001602150100003302166700002002199700002302219700002902242700002002271700003302291700002502324700002502349700002102374700001902395700002202414856012702436       2022                        eng d  a0981-942800aStacking herbicide detoxification and resistant genes improves glyphosate tolerance and reduces phytotoxicity in tobacco (Nicotiana tabacum L.) and rice (Oryza sativa L.)0 aStacking herbicide detoxification and resistant genes improves g  cOCT  a126-1380 v1893 a<p>
	Glyphosate residues retained in the growing meristematic tissues or in grains of glyphosate-resistant crops affect the plants physiological functions and crop yield. Removing glyphosate residues in the plants is desirable with no penalty on crop yield and quality. We report a new combination of scientific strategy to detoxify glyphosate that reduces the residual levels and improve crop resistance. The glyphosate detoxifying enzymes Aldo-keto reductase (AKR1) and mutated glycine oxidase (mGO) with different modes of action were co-expressed with modified EPSPS, which is insensitive to glyphosate in tobacco (Nicotiana tabacum L.) and rice (Oryza sativa L.). The transgenic tobacco plants expressing individual PsAKR1, mGO, CP4-EPSPS, combinations of PsAKR1:CP4EPSPS, PsAKR1:mGO, and multigene with PsAKR1: mGO: CP4EPSPS genes were developed. The bio-efficacy studies of in-vitro leaf regeneration on different concentrations of glyphosate, seedling bioassay, and spray on transgenic tobacco plants demonstrate that glyphosate detoxification with enhanced resistance. Comparative analysis of the transgenic tobacco plants reveals that double and multigene expressing transgenics had reduced accumulation of shikimic acid, glyphosate, and its primary residue AMPA, and increased levels of sarcosine were observed in all PsAKR1 expressing transgenics. The multigene expressing rice transgenics showed improved glyphosate resis-tance with yield maintenance. In summary, results suggest that stacking genes with two different detoxification mechanisms and insensitive EPSPS is a potential approach for developing glyphosate-resistant plants with less residual content.</p>
10aDetoxification10aGene stacking10aGlyphosate10aHerbicide10aMultigene10aResidual toxicity10aResistant10arice10aTobacco10aTransgenics1 aVennapusa, Amaranatha, Reddy1 aAgarwal, Subham1 aHm, Hanumanth, Rao1 aAarthy, Thiagarayaselvam1 aBabitha, K., C.1 aThulasiram, Hirekodathakallu1 aKulkarni, Mahesh, J.1 aMelmaiee, Kalpalatha1 aSudhakar, Chinta1 aUdayakumar, M.1 aVemanna, Ramu, S.  uhttps://library.ncl.res.in/content/stacking-herbicide-detoxification-and-resistant-genes-improves-glyphosate-tolerance-and