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

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.

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 515c-515
Author(s):  
Shanqiang Ke ◽  
Chiwon W. Lee

Coleoptile tissues from dark-germinated seedlings of Kentucky bluegrass (Poa pratensis L.) cv. Touchdown were excised and cultured on MS medium supplemented with 1.5-2.5 mg/liter picloram plus 0.2 mg/liter benzyladenine (BA) or with 4 mg/liter 2,4-D. Embryogenic calli were formed on media containing 1.5 mg/liter picloram plus 2.5 mg/liter 2,4-D in the dark. When these embryogenic calli were subcultured on MS medium containing either 0.15-0.3 mg/liter picloram or 0.2-0.5 mg/liter 2,4-D in a 16-h day/8-h night photoperiod, 10.5% of the cultures regenerated shoots. Pretreatment of cultures in the dark for 2 weeks prior to light exposure slightly increased the plant regeneration efficiency to 15.5%. Pigmentation of the regenerants varied with a ratio of 8.5 completely green: 2.5 green plus albino: 1 completely albino plants. The shoots were multiplied in the medium containing 0.5 mg/liter BA plus either 0.2 mg/liter picloram or 0.1 mg/liter indoleacetic acid (IAA). Over 90% cultures in the shoot proliferation medium produced roots after 4 weeks.


HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 616d-616 ◽  
Author(s):  
Shanqiang Ke ◽  
Chiwon W. Lee ◽  
Zong-Ming Cheng

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.


1993 ◽  
Vol 48 (3-4) ◽  
pp. 163-167
Author(s):  
Koichi Yoneyama ◽  
Yoshihiro Nakajima ◽  
Masaru Ogasawara ◽  
Hitoshi Kuramochi ◽  
Makoto Konnai ◽  
...  

Abstract Through the studies on structure-activity relationships of 5-acyl-3-(1-aminoalkylidene)-4-hydroxy-2 H-pyran-2,6(3 H)-dione derivatives in photosystem II (PS II) inhibition, overall lipophilicity of the molecule was found to be a major determinant for the activity. In the substituted N -benzyl derivatives, not only the lipophilicity but also the electronic and steric characters of the substituents greatly affected the activity. Their mode of PS II inhibition seemed to be similar to that of DCMU , whereas pyran-enamine derivatives needed to be highly lipophilic to block the electron transport in thylakoid membranes, which in turn diminished the permeability through biomembranes.


1979 ◽  
Vol 25 (10) ◽  
pp. 1197-1200 ◽  
Author(s):  
R. C. Shearman ◽  
W. L. Pedersen ◽  
R. V. Klucas ◽  
E. J. Kinbacher

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.


1981 ◽  
Vol 27 (1) ◽  
pp. 52-56 ◽  
Author(s):  
L. V. Wood ◽  
R. V. Klucas ◽  
R. C. Shearman

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.


1979 ◽  
Vol 59 (2) ◽  
pp. 469-473 ◽  
Author(s):  
R. G. INGRATTA ◽  
G. R. STEPHENSON ◽  
C. M. SWITZER

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.


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