کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8880007 | 1624789 | 2018 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Development of transgenic hairy roots and augmentation of secondary metabolites by precursor feeding in Sphagneticola calendulacea (L.) Pruski
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کلمات کلیدی
HPTLCAgrobacterium rhizogenesPGRAcetosyringoneMurashige and Skoog - Murashige و SkoogWedelolactone - WedloctoneBinary vector - بردار دودوییplant growth regulator - تنظیم کننده رشد گیاهHairy root - ریشه مودارfresh weight - وزن ترdry weight - وزن خشکPrecursor feeding - پیشگیر تغذیهHigh performance thin-layer chromatography - کروماتوگرافی لایه نازک با عملکرد بالاhigh performance liquid chromatography - کروماتوگرافی مایع با کارایی بالاHPLC - کروماتوگرافی مایعی کارا
موضوعات مرتبط
علوم زیستی و بیوفناوری
علوم کشاورزی و بیولوژیک
علوم زراعت و اصلاح نباتات
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Sphagneticola calendulacea (L.) Pruski [synonym Wedelia chinensis (Osbeck) Merrill] is an important medicinal plant that possesses a variety of bioactive secondary metabolites. A stable transgenic hairy root culture has been developed using engineered Agrobacterium rhizogenes strain LBA1334 harboring pCAM:2â¯Ãâ¯35S:gusA binary vector, to achieve high yield of secondary metabolites. Putative transgenic hairy roots appeared with highest transformation frequency (66.67%) when leaf explants were immersed in A. rhizogenes suspension for 10â¯min and co-cultivated for three days. Incorporation of foreign genes was confirmed by initial GUS staining assay followed by polymerase chain reaction analysis of rolA, rolB, rolC and gusA genes. Time course study revealed that the transgenic hairy roots grew rapidly in ½ Murashige and Skoog liquid medium with highest biomass production of 0.61â¯g per 50â¯ml dry weight after 28â¯days of culture. In the precursor feeding experiment, the biomass of hairy roots was further increased to 0.698â¯g per 50â¯ml dry weight when 0.5â¯mM phenylalanine was added to the 10â¯days old growing culture. High Performance Thin Layer Chromatography showed that the transgenic hairy roots accumulated enhanced level of wedelolactone (422.01â¯Î¼gâ¯gâ1 dry weight), which was 1.43 and 1.37 times higher than those of in vivo (294.44â¯Î¼gâ¯gâ1 dry weight) and in vitro-grown (308.28â¯Î¼gâ¯gâ1 dry weight) plantlets, respectively. Precursor feeding with phenylalanine further augmented the accumulation of wedelolactone to 440.33â¯Î¼gâ¯gâ1 dry weight in the transgenic hairy roots. In addition, the content of several important phenolic and flavonoid compounds were increased in the transgenic hairy roots as analyzed by High Performance Liquid Chromatography. The protocol offers a scope to introduce foreign genes and further regulate the production of secondary metabolites.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Industrial Crops and Products - Volume 121, 1 October 2018, Pages 206-215
Journal: Industrial Crops and Products - Volume 121, 1 October 2018, Pages 206-215
نویسندگان
Suprabuddha Kundu, Umme Salma, Md. Nasim Ali, Alok Kumar Hazra, Nirmal Mandal,