Gene expression of near-isogenic lines (NILs) carrying blast resistance genes Pi9 and Pi54 in the background of rice cultivar Mushk Budji.

BACKGROUND: Kashmir valley, India is a homeland to rice landraces like Zag, Nunbeoul, Qadirbeigh, Kawkadur, Kamad, Mushk Budji, etc., generally characterized by short grains, aroma, earliness and cold tolerance. Mushk Budji is a commercially important speciality rice known for its taste and aroma, nonetheless, is extremely vulnerable to blast disease. Through the use of the marker-assisted backcrossing (MABC) approach, a set of 24 Near-isogenic lines (NILs) was created, and the lines with the highest background genome recovery were chosen. The expression analysis was carried out for the component genes and other eight pathway genes related to blast resistance. RESULTS: The major blast resistance genes Pi9 (from IRBL-9W) and Pi54 (from DHMAS 70Q 164-1b) were incorporated following simultaneous-but-step-wise MABC. The NILs harbouring genes Pi9 + Pi54, Pi9 and Pi54 expressed resistance to isolate (Mo-nwi-kash-32) under controlled and natural field conditions. The loci controlling ETI (effector triggered immunity) included the gene Pi9 and showed 61.18 and 60.27 fold change in relative gene expression in Pi54 + Pi9 and Pi9 carrying NILs against RP Mushk Budji. Pi54 was up regulated and showed 41 and 21 fold change in relative gene expression for NIL-Pi54 + Pi9 and NIL-Pi54, respectively. Among the pathway genes, LOC_Os01g60600 (WRKY 108) recorded 8 and 7.5 fold up regulation in Pi9 and Pi54 NILs. CONCLUSION: The NILs showed recurrent parent genome recovery (RPG) per cent of 81.67 to 92.54 and were on par in performance to recurrent parent Mushk Budji. The lines were utilized to study the expression of the loci controlling WRKYs, peroxidases and chitinases that confer overall ETI response.

LC-MS guided isolation, quantification and antioxidant evaluation of bioactive principles from Epimedium elatum.

This article presents the isolation, quantification and antioxidant evaluation of bioactive principles from Epimedium elatum. LC-MS guided isolation technique was applied for the separation of target constituents. Three isolates; magnoflorine, chrysin and dibenzylideneacetone (DBA) were isolated for the first time from E. elatum using LC-MS guided isolation method. Nine natural products, viz. icariin, epimedoside A, epimedin A, epimedin B, epimedin C, ikarisoside C, baohuoside II, magnoflorine and chrysin were simultaneously quantified by reverse phase HPLC-UV-DAD method. The HPLC method was validated in terms of precision and accuracy. Excellent specificity and linearity within test ranges for all standard calibration curves having regression coefficient of different linear equations in the range of 0.9966-0.9999 were observed. Relative recovery rates varied between 98.09±0.44 and 105.34±1.89% with relative standard deviation of less than 3%. This modified HPLC method is in accordance with yinyanghuo. All the 10 isolated constituents were screened for DPPH radical scavenging activity. Dibenzylideneacetone (DBA) turned out to be the most potent isolate with IC(50) of 4.32 µM.