کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5762974 | 1625147 | 2017 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Optimization of transient GUS expression of Agrobacterium-mediated transformation in Dierama erectum Hilliard using sonication and Agrobacterium
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کلمات کلیدی
NaACaMV35SCauliflower Mosaic Virus 35S promoterGUSIBACTAB1-naphthaleneacetic acid - 1-نفتالین اسیدهای اسید2,4-D - 2،4-D2,4-dichlorophenoxyacetic acid - 2،4-dichlorophenoxyacetic اسیدMurashige and Skoog - Murashige و Skoogβ-glucuronidase - β-گلوکورونیدازcetyltrimethylammonium bromide - اتلی تریمتیل آمونیوم برومیدindole-3-butyric acid - اسید Indole-3-ButyricGene delivery - انتقال ژن Genetic transformation - تحول ژنتیکیSonication - سونیکیشن SEM - مدل معادلات ساختاری / میکروسکوپ الکترونی روبشیscanning electron microscope - میکروسکوپ الکترونی اسکنpolymerase chain reaction - واکنش زنجیره ای پلیمرازPCR - واکنش زنجیرهٔ پلیمرازoptical density - چگالی نوریSAAT - کی
موضوعات مرتبط
علوم زیستی و بیوفناوری
علوم کشاورزی و بیولوژیک
علوم زراعت و اصلاح نباتات
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Efficient Agrobacterium-mediated transformation was established via sonication of embryonic shoot apical meristems (ESAMs) of Dierama erectum Hilliard. Effects of explant types, co-cultivation time, acetosyringone concentration, Agrobacterium concentration and different gene delivery methods were evaluated for higher efficiency of genetic transformation in D. erectum. An explant type (ESAMs), concentration of Agrobacterium inoculum (OD600 of 1.6) and acetosyringone (50 mg Lâ 1), and co-cultivation duration (3 days) were optimized for efficient genetic transformation of D. erectum. The transformation efficiency varied with explant types (from 0 to 60%), concentrations of bacteria (10 to 55%) and acetosyringone (50 to 90%) and period of co-cultivation (30 to 70%). The transformation efficiency was best with ESAMs explants compared with callus clusters. The gene delivery method via sonication-assisted Agrobacterium-mediated transformation (SAAT) provided higher transformation efficiency (40%) GUS expression compared with agrobacterial monolayer and agrobacterial suspension which gave less than 5% transformation efficiency. The putative transgenic plants which histochemically expressed GUS, were confirmed further with PCR and 35.3% of the plants were GUS positive. Stable integration of the transgene was not demonstrated hence the GUS expression observed was regarded as transient. This newly developed transformation system may facilitate improvement of D. erectum characteristics and other related geophytes.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: South African Journal of Botany - Volume 111, July 2017, Pages 307-312
Journal: South African Journal of Botany - Volume 111, July 2017, Pages 307-312
نویسندگان
M.J. Koetle, P. Baskaran, J.F. Finnie, V. Soos, E. Balázs, J. Van Staden,