scholarly journals Genetic Transformation of Kentucky Bluegrass with the rolC Gene

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 616d-616 ◽  
Author(s):  
Shanqiang Ke ◽  
Chiwon W. Lee ◽  
Zong-Ming Cheng
Keyword(s):  
Southern Hybridization ◽  
Poa Pratensis ◽  
Kentucky Bluegrass ◽  
Naphthaleneacetic Acid ◽  
Gus Assay ◽  
Similar Frequency ◽  
Rolc Gene ◽  
Putative Transformants ◽  
Regenerated Plants ◽  
Npt Ii

Coleoptile tissues excised from young seedlings of `Touchdown' Kentucky bluegrass (Poa pratensis L.) were bombarded with the disarmed Agrobacterium tumefaciens strain EHA 101 carrying rolC (from A. rhizogenes), NPT II and GUS genes. These tissues were then cultured on Murashige and Skoog (MS) medium containing 0.2 mg·L–1 picloram, 0.01 mg·L–1 naphthaleneacetic acid (NAA), 150 mg·L–1 kanamycin, and 50 m acetosyringone. Calli formed on this medium within 2 weeks. The regenerated plants from these calli were analyzed for the presence of the GUS and rolC genes by histochemical GUS assay, PCR, and Southern hybridization. Only 3.7% of the regenerants were transformed when determined by the GUS assay. A similar frequency of transformation in the regenerated plants was obtained after bombarding the coleoptile tissues with the DNA isolated from the pGA-GUSGF-rolC plasmid. Most of the putative transformants were either albinos or variegated plants that are composed of both albino and green tissues.

scholarly journals Influence of the rolC Gene on Proteins Associated with Stroma and Thylakoid Membranes of Chloroplasts in Transgenic Plants of Kentucky Bluegrass

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 482D-482
Author(s):  
Shanqiang Ke ◽  
Chiwon W. Lee ◽  
Murray E. Duysen
Keyword(s):  
Transgenic Plants ◽  
Poa Pratensis ◽  
Kentucky Bluegrass ◽  
Thylakoid Membranes ◽  
Induction Medium ◽  
Protein Accumulation ◽  
Naphthaleneacetic Acid ◽  
Plastid Development ◽  
Ps Ii ◽  
Albino Plants

The effects of the expression of the rolC gene on protein accumulation in the chloroplasts of transgenic Kentucky bluegrass (Poa pratensis L.) were investigated. Coleoptile tissues excised from 3-day dark-grown seedlings were bombarded with tungsten particles coated with DNA of the engineered plasmid, pGA-GUSGF, containing the npt II, gus, and rolC genes. The tissues were cultured on callus induction medium, which consists of MS salts supplemented with 0.2 mg/L picloram, 0.01 mg/L naphthaleneacetic acid (NAA) 250 mg/L kanamycin, and 100 mM acetosyringone. The putative transformants were either albinos or variegated plants composed of white and green sections. These albino plants had little or no stroma-based 56-kDa and 14-kDa subunits of the suspected Rubisco proteins, which are expressed in response to genes in the nucleus and plastid, respectively. The albino plants also lacked the 110-kDa and 57–58-kDa, and 43, 47-kDa polypeptides in PS I, coupling factor, and PS II in thylakoid membranes, respectively. These proteins involved in photosynthesis are translated from plastidbased genes. No light-harvesting complex proteins (LHC) were observed in these albino plants. LHC genes are encoded in the nucleus. The thylakoid membrane proteins in the chloroplasts of the rolC transgenic variegated plants contained these proteins. Our data suggest that the nucleus and plastid gene products for plastid development are concomitantly impaired by expression of genes in the transgenic plants.

Nitrogen fixation associated with 'Park' Kentucky bluegrass (Poapratensis L.)

1979 ◽  
Vol 25 (10) ◽  
pp. 1197-1200 ◽  
Author(s):  
R. C. Shearman ◽  
W. L. Pedersen ◽  
R. V. Klucas ◽  
E. J. Kinbacher
Keyword(s):  
Nitrogen Fixation ◽  
Acetylene Reduction ◽  
Poa Pratensis ◽  
Kentucky Bluegrass ◽  
Acetylene Reduction Activity ◽  
Controlled Environment ◽  
Nitrogen Accumulation ◽  
Reduction Activity ◽  
Significant Difference ◽  
Reduction Assay

Associative nitrogen fixation in Kentucky bluegrass (Poa pratensis L.) turfs inoculated with five nitrogen-fixing bacterial isolates was evaluated using the acetylene reduction assay and nitrogen accumulation as indicators of fixation. 'Park' and 'Nugget' Kentucky bluegrass turfs were grown in controlled environment chambers and inoculated with Klebsiella pneumoniae (W-2, W-6, and W-14), Erwinia herbicola (W-8), and Enterobacter cloacae (W-11). 'Park' inoculated with K. pneumoniae (W-6) had significant acetylene reduction activity using undisturbed turfs. Other treatments including turfs treated with heat-killed cells had no significant difference in acetylene reduction. In a second study, 'Park' and 'South Dakota Certified' turfs were grown in a greenhouse and inoculated with K. pneumoniae (W-6) and E. herbicola (W-8). 'Park' inoculated with K. pneumoniae (W-6) had increased acetylene reduction activity rates and also a greater nitrogen accumulation in aerial tissues when compared to controls. Acetylene reduction activity was correlated (r = 0.92) to nitrogen accumulation. Other treatments did not effectively increase acetylene reduction activity or nitrogen accumulation.

Nitrogen fixation (acetylene reduction) by Klebsiella pneumoniae in association with 'Park' Kentucky bluegrass (Poa pratensis L.)

1981 ◽  
Vol 27 (1) ◽  
pp. 52-56 ◽  
Author(s):  
L. V. Wood ◽  
R. V. Klucas ◽  
R. C. Shearman
Keyword(s):  
Nitrogen Fixation ◽  
Klebsiella Pneumoniae ◽  
Ethylene Production ◽  
Acetylene Reduction ◽  
Nitrogenase Activity ◽  
Poa Pratensis ◽  
Kentucky Bluegrass ◽  
Surface Sterilization ◽  
Reducing Activity ◽  
Acetylene Reducing Activity

Turfs of 'Park' Kentucky bluegrass reestablished in the greenhouse and inoculated with Klebsiella pneumoniae (W6) showed significantly increased nitrogen fixation (acetylene reduction) compared with control turfs. Mean ethylene production rates per pot were 368 nmol h−1 for K. pneumoniae treated turfs, 55 nmol h−1 for heat-killed K. pneumoniae treated turfs, and 44 nmol h−1 for untreated turfs. Calculated lag periods before activity was observed were generally very short (less than 1 h).When 'Park' Kentucky bluegrass was grown from seed on soil-less medium of Turface, a fired aggregate clay, inoculation with K. pneumoniae (W6) resulted in 9 of 11 turfs showing nitrogenase activity (mean ethylene producion rate per pot was 195 nmol h−1). Only 3 of 11 turfs treated with heat-killed K. pneumoniae showed any activity and their mean rate of ethylene production (40 nmol h−1 per pot) was significantly lower than that for turfs treated with K. pneumoniae.Using the 'Park'–Turface soil-less model system it was shown that acetylene reducing activity was (i) root associated, (ii) generally highest at a depth of 1–4 cm below the surface, (iii) enhanced by washing excised roots, and (iv) inhibited by surface sterilization of excised roots. Klebsiella pneumoniae was recovered from Turface and roots showing acetylene reducing activity.

SELECTIVE CONTROL OF ANNUAL BLUEGRASS IN KENTUCKY BLUEGRASS TURF WITH LINURON

1979 ◽  
Vol 59 (2) ◽  
pp. 469-473 ◽  
Author(s):  
R. G. INGRATTA ◽  
G. R. STEPHENSON ◽  
C. M. SWITZER
Keyword(s):  
Temperature Regime ◽  
Poa Pratensis ◽  
Kentucky Bluegrass ◽  
Controlled Environment ◽  
Night Temperature ◽  
Annual Bluegrass ◽  
Selective Control ◽  
Excellent Control

Optimum top growth of annual bluegrass (Pao annua L.) and Kentucky bluegrass (Poa pratensis L.) was obtained at 24/12 °C day/night temperature regime in controlled environment studies. The tolerance of seedling Kentucky bluegrass to linuron [3-(3,4-dichlorophenyl)-1)methylurea] appeared to be greatest at this temperature regime when photoperiods were 16 h in length. A granular formulation of linuron gave excellent control of annual bluegrass in Kentucky bluegrass turf at 6.7 kg/ha when applied postemergence. At this rate, all culitivars of Kentucky bluegrass tested, with the exception of Fylking, were tolerant to linuron as a granular formulation. After application of linuron at 3.4–6.7 kg/ha, phytotoxic residues remained in the soil at sufficient levels to injure seedling Kentucky bluegrass for up to 3 mo.

scholarly journals Improved the Agrobacterium tumefaciens-mediated transformation of cucumber by a modified the using of antibiotics and acetosyringone

2020 ◽  
Author(s):  
Li'ang Chai ◽  
Changxia Du ◽  
Huaifu Fan ◽  
Chen Liu ◽  
Yuyang Si
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Culture Medium ◽  
Early Stage ◽  
Marker Gene ◽  
Vegetable Crops ◽  
Regenerated Plants ◽  
Npt Ii ◽  
Screening Efficiency ◽  
Speed Up ◽  
Polymerase Chain

Abstract Background: Cucumber (Cucumis sativus) is one of the most important vegetable crops in the world. As conventional breeding of cucumber is very challenging, genetic engineering is an alternative option to introduce important traits such as enhanced stress resistance and nutritional value. However, the efficiency of the transformation system depends on genotypes, transformation conditions, selection agents, etc. This study aims to speed up the process of Agrobacterium-mediated transformation of cucumber. ‘ Xintai mici ’, a very popular and typical north China-type cucumber variety, was transformed with Agrobacterium GV3101. The strain carried pCAMBIA2300s plasmid, a double vector with the marker gene of neomycin phosphotransferase II ( npt II). Results: The research results indicated that cefotaxime sodium was suitable for inhibiting Agrobacterium in the stage of screening and bud elongation. Timentin was best used during rooting stage. Furthermore, 25 mg/L kanamycin was used in the early stage of screening and increased to 50 mg/L for further screening. At the bud elongation and rooting stage, 75 and 100 mg/L kanamycin was used respectively to improve the screening efficiency. In order to obtain the highest regeneration frequency of resistant buds, 50, 150, and 100 μM acetosyringone were added in the pre-culture medium, infection solution, and co-culture medium respectively. To confirm the presence of the transgenes, DNA from npt II transgenic cucumber plants was analyzed by polymerase chain reaction after transplanting resistant regenerated plants. Conclusions: We finally achieved an 8.1% conversion, which was among the highest values reported until date using cucumber ‘ Xintai mici ’. Thus an effective protocol for Agrobacterium tumefaciens -mediated genetic transformation of cucumber was optimized.

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