BackgroundIn soybean somatic embryo transformation, the standard selection agent currently used is hygromycin. It may be preferable to avoid use of antibiotic resistance genes in foods. The objective of these experiments was to develop a selection system for producing transgenic soybean somatic embryos without the use of antibiotics such as hygromycin.ResultsWhen tested against different alternate selection agents our studies show that 0.16 μg/mL glufosinate, 40 mg/L isopropylamine-glyphosate, 0.5 mg/mL (S-(2 aminoethyl)-L-cysteine) (AEC) and the acetolactate synthase (ALS) inhibitors Exceed® and Synchrony® both at 150 μg/mL inhibited soybean somatic embryo growth. Even at the concentration of 2 mg/mL, lysine+threonine (LT) were poor selection agents. The use of AEC may be preferable since it is a natural compound. Unlike the plant enzyme, dihydrodipicolinate synthase (DHPS) from E. coli is not feed-back inhibited by physiological concentrations of lysine. The dapA gene which codes for E. coli DHPS was expressed in soybean somatic embryos under the control of the CaMV 35S promoter. Following introduction of the construct into embryogenic tissue of soybean, transgenic events were recovered by incubating the tissue in liquid medium containing AEC at a concentration of 5 mM. Only transgenic soybeans were able to grow at this concentration of AEC; no escapes were observed.ConclusionGenetically engineered soybeans expressing a lysine insensitive DHPS gene can be selected with the non-antibiotic selection agent AEC. We also report here the inhibitory effects of glufosinate, (isopropylamine-glyphosate) (Roundup®), AEC and the ALS inhibitors Exceed® and Synchrony® against different tissues of soybean
Three varieties of Pea (Pimm sativum L.) viz. EC33866 (early). P-i 16 (medium) and T-163 (late), differing in their flowering and maturity times, were taken for this study. The early variety recorded lower dry matter, COj exchange rate and lower yield. The harvest index (HI) was the highest (44 %) in this variety. Though medium and late varieties did not differ significantly in dry matter accumulation and pod yield, they diflered in their HI. The HI in the medium variety is 42 % but in the late variety it is only 36%.The late variety maintained high leaf area, leaf dry weight, high shoot/root, shoot/nodule and root/nodule ratios, increased nodule dry weight and maximum glycolate oxidase activity. But acetylene reduction activity was low, and the harvest index was lowest. On the other hand, in the early variety the dry matter accumulation was lowest but Hi was highest. The major reasons for high HI in this variety were translocation of photosynthates to reproductive organs and lower photorespiration as indicated by low glycolate oxidase activity.
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