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1.
JARI P T VALKONEN 《The Annals of applied biology》1993,123(2):309-314
Plants from 2nd to 6th year leys of the legume goat's rue (Galega orientalis Lam.) were tested for infection with bean yellow mosaic (BYMV), bean common mosaic (BCMV), alfalfa mosaic (AMV), broad bean stain (BBSV), red clover mottle (RCMV) and cucumber mosaic (CMV) viruses by enzyme-linked immunosorbent assay (ELISA), electron microscopy, and by sap-inoculation to various test plant species. No virus infections were observed in goat's rue in the field. Glasshouse-grown seedlings of goat's rue were inoculated with the above viruses. No virus was detected in the inoculated plants. The results suggest that goat's rue is extremely resistant to the above six viruses which are important in other forage legumes. 相似文献
2.
Fungi and Gram-negative bacteria as soilborne minor pathogens of goat's rue (Galega orientalis Lam.)
JARI P T VALKONEN WIPATORN von HEIROTH MERVI-LEENA SAVELA 《The Annals of applied biology》1993,123(2):257-269
Fungi and Gram-negative bacteria were isolated from inside the roots of field-grown goat's rue (Galega orientalis). Fungi were isolated from three plants out of a total of 45 tested. Two multinuclear Rhizoctonia solani isolates were identified to the anastomosis group 5 (R. solani AG-5-Gal) using pairings with known AG test cultures. One fungal isolate was identified to Phoma chrysanthemicola. Gram-negative bacteria were isolated from three plants out of 25 tested. They were identified using classical methods, the BIOLOG identification system based on the utilisation of 95 different carbon sources, and the MIDI system for the analysis of whole cell fatty acids. The two latter systems were computer-associated and utilised an extensive reference library of isolates. One bacterial isolate was identified as Enterobacter agglomerans and two isolates as Pseudomonas marginalis. R. solani AG-5-Gal reduced the emergence of Lupinus luteus, L. polyphyllus and french bean (Phaseolus vulgaris) and the growth of broad bean (Viciafaba), L. luteus and french bean, but did not cause obvious damage in goat's rue and pea (Pisum sativum). However, R. solani AG-5-Gal was re-isolated from the roots of all the test plant species following inoculation. P. chrysanthemicola reduced the emergence of L. polyphyllus and the growth of goat's rue, french bean and broad bean, and it was re-isolated from all of the test plant species (except for french bean) following inoculation. All the bacteria reduced the emergence of french bean, but not that of goat's rue and pea, when applied to the soil. When the roots were dipped into bacterial suspension, all the bacteria damaged french bean and L. polyphyllus. Additionally, P. marginalis JV3 damaged goat's rue and red clover. The pathogenicity of the fungi and bacteria were not changed when they were double-inoculated in pairs, except for R. solani AG-5-Gal and P. marginalis JV2 which reduced the emergence of goat's rue when inoculated together but not when inoculated separately. 相似文献
3.
Genetic transformation of Cymbidium orchid by particle bombardment 总被引:13,自引:0,他引:13
J. Yang H.-J. Lee D. H. Shin S. K. Oh J. H. Seon K. Y. Paek K.-H. Han 《Plant cell reports》1999,18(12):978-984
A protocol is presented for genetically engineering Cymbidium orchid using particle bombardment. This protocol enabled the routine transformation of orchid plants that were previously
difficult to transform. Liquid culture was used to generate a large number of protocorm-like bodies (PLBs) to be bombarded
and to promote continued development of the bombarded meristematic tissue. Plasmid DNA (pKH200) carrying the GUS-INT and NPTII
genes flanked by tobacco matrix attachment regions was introduced into the meristematic cells of PLBs by particle acceleration.
The transformed PLBs were proliferated and selected for kanamycin resistance conferred by the introduced NPTII gene. Shoot
regeneration was then induced from the kanamycin-resistant PLBs, and transgenic plantlets were produced. Both the kanamycin-resistant
PLBs and regenerated shoots expressed the GUS-INT gene. The presence of the introduced gene in the transformed orchid plants
was confirmed by PCR analysis, sequencing and Southern blot analysis of the PCR product. The recovered transgenic plants were
established in soil and acclimatized in the greenhouse.
Received: 20 July 1998 / Revision received: 2 December 1998 / Accepted: 17 December 1998 相似文献
4.
An efficient protocol for Agrobacterium tumefaciens-mediated transformation of four commercial cultivars of Brassica oleracea var. capitata is described. A strain of A. tumefaciens LBA4404 with the neomycin phosphotransferase gene (nptII) and a CaMV 35S-peroxidase gene cassette were used for co-cultivation. Preliminary selection of regenerated transgenic plants
was performed on kanamycin-containing medium. The frequency of transgenic plants was calculated on the basis of GUS (β-glucuronidase)
activity detected by the histochemical X-gluc test. Tissue-specific GUS expression driven by the peroxidase gene promoter
in transgenic plants was analysed by GUS staining. The transformation rates of the commercial cultivars of B. oleracea was higher than in previous reports. Southern blot analysis revealed that integration of marker genes occurred in single
and multiple loci in the genome. All transgenic plants grew normally after a brief vernalization period and showed stable
inheritance of the marker gene. The present study demonstrates that morphologically normal, fertile transgenic plants of B. oleracea can be obtained.
Received: 24 August 1999 / Revision received: 23 November 1999 / Accepted: 3 December 相似文献
5.
An Agrobacterium tumefaciens-mediated transformation method has been developed for onions (Allium cepa L.) using immature embryos as the explant source. Transgenic plants were recovered from the open-pollinated onion cultivar
Canterbury Longkeeper at a maximum transformation frequency from immature embryos of 2.7%. The method takes between 3–5 months
from explant to primary regenerant entering the glasshouse. Multiple-shoot formation from primary transgenic material made
possible the clonal multiplication of transformants. The binary vector used carried the nptII antibiotic resistance gene and the m-gfp5-ER reporter gene. Transgenic cultures were initially screened for their ability to fluoresce and to grow in the presence of
geneticin (5–25 mg/l). The transgenic nature of individual plants was confirmed by Southern blot analysis.
Received: 12 October 1998 / Revision received: 17 May 1999 Accepted: 14 June 1999 相似文献
6.
An effective method has been developed for the stable transformation and regeneration of Cavendish banana (Musa spp. AAA group) cv 'Grand Nain' by microprojectile bombardment. Embryogenic cell suspensions were initiated using immature
male flowers as the explant. Cells were co-bombarded with the neomycin phosphotransferase (nptII) selectable marker gene under the control of a banana bunchy top virus (BBTV) promoter or the CaMV 35S promoter, and either
the β-glucuronidase (uidA) reporter gene or BBTV genes under the control of the maize polyubiquitin promoter. Plants were regenerated, under selection
with kanamycin, that were co-transformed with nptII and either the uidA or BBTV genes. Molecular characterisation of transformants demonstrated that the transgenes had been stably integrated into
the banana genome.
Received: 22 June 1998 / Revision received: 29 March 1999 / Accepted 1 May 1999 相似文献
7.
K. V. Krishnamurthy K. Suhasini A. P. Sagare M. Meixner A. de Kathen T. Pickardt O. Schieder 《Plant cell reports》2000,19(3):235-240
Embryo axes of four accessions of chickpea (Cicer arietinum L.) were treated with Agrobacterium tumefaciens strains C58C1/GV2260 carrying the plasmid p35SGUSINT and EHA101 harbouring the plasmid pIBGUS. In both vectors the GUS gene
is interrupted by an intron. After inoculation shoot formation was promoted on MS medium containing 0.5 mg/l BAP under a selection
pressure of 100 mg/l kanamycin or 10 mg/l phosphinothricin, depending on the construct used for transformation. Expression
of the chimeric GUS gene was confirmed by histochemical localization of GUS activity in regenerated shoots. Resistant shoots
were grafted onto 5-day-old dark-grown seedlings, and mature plants could be recovered. T-DNA integration was confirmed by
Southern analysis by random selection of putative transformants. The analysis of 4 plantlets of the T1 progeny revealed that
none of them was GUS-positive, whereas the presence of the nptII gene could be detected by polymerase chain reaction.
Received: 30 May 1997 / Revision received: 18 September 1997 / Accepted: 22 March 1999 相似文献
8.
Factors that influence Agrobacterium rhizogenes-mediated transformation of broccoli (Brassica oleracea L. var. italica) 总被引:2,自引:0,他引:2
An improved broccoli transformation system was developed by optimising several factors that affect the rate of effective
Agrobacterium-mediated transformation. Leaf explants of cultivar Shogun were co-cultivated with Agrobacterium rhizogenes strain A4T harbouring the binary vector pART278. The T-DNA of this binary vector contains a neomycin phosphotransferase II
(NOS-NPTII-NOS) gene for kanamycin resistance and a β-glucuronidase (35S-GUS-OCS) gene. Several media and factors were evaluated
including combinations of arginine, mannopine, acetosyringone and the use of feeder cell layers. The new protocol includes
the use of 200 μm acetosyringone in LB medium for bacterial growth, the use of a Brassica campestris feeder cell layer, 10 mm mannopine and 50 μm acetosyringone in the co-cultivation medium and 1 mm arginine in the selection medium. The use of this optimised protocol produced transformation rates of 33% in preliminary
experiments transforming broccoli with the antisense 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase gene from pTOM13.
Received: 2 July 1998 / Revision received: 9 February 2000 / Accepted: 17 February 2000 相似文献
9.
An efficient wheat transformation procedure: transformed calli with long-term morphogenic potential for plant regeneration 总被引:9,自引:0,他引:9
L. Zhang J. J. Rybczynski W. G. Langenberg A. Mitra R. French 《Plant cell reports》2000,19(3):241-250
A method for producing large numbers of transgenic wheat plants has been developed. With this approach, an average of 9.7%
of immature embryo explants were transformed and generated multiple self-fertile, independently transformed plants. No untransformed
plants, or escapes, were regenerated. This transformation procedure uses morphogenic calli derived from scutellum tissue of
immature embryos of Triticum aestivum cv. Bobwhite co-bombarded with separate plasmids carrying a selectable marker gene (bar) and a gene of interest, respectively. Transformed wheat calli with a vigorous growth phenotype were obtained by extended
culture on media containing 5.0 mg/l bialaphos. These calli retained morphogenic potential and were competent for plant regeneration
for as long as 11 months. The bar gene and the gene of interest were co-expressed in T0 progeny plants. This wheat transformation protocol may facilitate quantitative
production of multiple transgenic plants and significantly reduce the cost and labor otherwise required for screening out
untransformed escapes.
Received: 15 June 1998 / Revision received: 6 April 1999 / Accepted: 26 April 1999 相似文献
10.
Recovery of transgenic rice plants expressing the rice dwarf virus outer coat protein gene (S8) 总被引:3,自引:0,他引:3
H. H. Zheng Y. Li Z. H. Yu W. Li M. Y. Chen X. T. Ming R. Casper Z. L. Chen 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,94(3-4):522-527
The coding region of the eighth largest segment (S8) of the rice dwarf virus (RDV) was obtained from a RDV Fujian isolate.
It was then cloned into pTrcHisA for expression in E. coli and into vector pE3 for plant transformation. By using callus derived from mature rice embryos as the target tissue, we obtained
regenerated rice plants after bombardment of the former with plasmid pE3R8 containing the RDV S8 gene and the marker gene
neomycin phosphotransferase (NPT II). Southern blotting confirmed the integration of the RDV S8 gene into the rice genome.
The expression of the outer coat protein in both E. coli and rice plants was confirmed by western blotting. The recovery of transgenic rice plants expressing S8 gene is an important
step towards studying the function of the RDV genes and obtaining RDV-resistant rice plants.
Received: 1 March 1996 / Accepted: 2 August 1996 相似文献
11.
We describe a protocol for Agrobacterium tumefaciens-mediated transformation of hybrid cottonwoods (Populus sections Tacamahaca Spach. and Aigeiros Duby). The protocol has allowed routine transformation of several economically important
cottonwood hybrids (Populus trichocarpa Torr. & Gray×P. deltoides Bartr. ex. Marsh. and P. deltoides×P. nigra L.) that were previously difficult to transform. The procedure was applied to 11 different hybrid cottonwood genotypes and
one P. deltoides genotype using kanamycin as the selection agent. Additional experiments showed a very strong interaction between auxin preculture
and the effectiveness of various cytokinins for induction of shoot organogenesis. The data also demonstrated the superiority
of Agrobacterium strain EHA105 over C58 and LBA4404 for T-DNA transfer based on transient assays with a reporter gene.
Received: 16 June 1998 / Revision received: 5 February 1999 / Accepted: 14 April 1999 相似文献
12.
The report describes a system for somatic embryogenesis and direct plant regeneration from the embryos of Manihot glaziovii. Somatic embryos were obtained by culturing young leaf lobes (3–6 mm long) adjacent to the apex in Murashige and Skoog medium
containing 18 μm 2,4-dichlorophenoxy acetic acid for 20 days and then transferring them to a maturation medium with 0.5 μm 6-benzylaminopurine. Secondary embryogenesis was induced from cotyledonary segments of somatic embryos by using the same
protocol as that for primary embryogenesis. For regeneration, somatic embryos were cultured in medium supplemented with 10−4 m kinetin and 53.4% of them developed into plantlets. Linamarin and linamarase were not detected in calli or in somatic embryos.
Linamarin content was found to be highest in leaves of regenerated plantlets, followed by stem and root tissues. Levels of
linamarase activity were almost the same in leaves and stem tissues and very low in roots.
Received: 19 April 1999 / Revision received: 11 August 1999 / Accepted: 17 August 1999 相似文献
13.
The efficiency of several promoters (pin2 from potato, ubiquitin from sunflower, rolC from Agrobacterium rhizogenes, act1 from rice and CaMV 35S from cauliflower mosaic virus) fused to the uidA reporter gene was measured after biolistic bombardment of birch leaves (Betula pendula L.). The highest level of β-glucuronidase (GUS) activity was achieved with the pin2 promoter and the lowest activity with the CaMV 35S promoter. The activity of the potato wound-inducible promoter (pin2) was also tested in stably transformed birch. The promoter showed induced activity after mechanical wounding and feeding
by leaf weevils. The systemic effect was confirmed by enhanced GUS activity in non-wounded leaves. The results of this study
indicated that the potato wound-inducible promoter maintains its function in birch and would be a suitable promoter in studies
of insect-birch interaction at the molecular level.
Received: 17 October 1996 / Revision received: 7 February 1997 / Accepted: 1 March 1997 相似文献
14.
Agrobacterium-mediated transformation and plant regeneration from hypocotyl segments of Japanese persimmon (Diospyros kaki Thunb) 总被引:1,自引:0,他引:1
Hypocotyl segments from the seeds of Japanese persimmon (Diospyros kaki Thunb) were cultured on a modified Murashige and Skoog medium supplemented with N-(2-chloro-4-pyridyl)-N′-phenylurea, zeatin
or 6-benzylaminopurine. The highest frequency of shoot regeneration was observed when the segments were cultured on medium
containing 2 mg/l of zeatin. This culture system was adapted to Agrobacterium-mediated transformation. The hypocotyl segments were inoculated with Agrobacterium tumefaciens strains harboring binary vectors, which contained the neomycin phosphotransferase II gene and the β-glucuronidase gene. Regenerated shoots were selected on a medium containing kanamycin. Histochemical GUS assay showed that
the shoots regenerated from the segments inoculated with EHA101/pSMAK251 expressed the gus gene. The presence and integration of the gus gene was confirmed by polymerase chain reaction (PCR) and Southern blot analysis. The regeneration frequency of transformed
shoot was 11.1%. The transgenic shoots were rooted and developed into whole plants within 4–5 months.
Received: 18 August 1997 / Revision received: 8 October 1997 / Accepted: 11 November 1997 相似文献
15.
A procedure for producing transgenic Chinese cabbage plants by inoculating cotyledonary explants with Agrobacterium tumefaciens strain EHA101 carrying a binary vector pIG121Hm, which contains kanamycin-resistance and hygromycin-resistance genes and
the GUS reporter gene, is described. Infection was most effective (highest infection frequency) when explants were infected
with Agrobacterium for 15 min and co-cultivated for 3 days in co-cultivation medium at pH 5.2 supplemented with 10 mg/l acetosyringone. Transgenic
plants of all three cultivars used were obtained with frequencies of 1.6–2.7% when the explants were regenerated in shoot
regeneration medium solidified with 1.6% agar. A histochemical GUS assay and PCR and Southern blot analyses confirmed that
transformation had occurred. Genetic analysis of T1 progeny showed that the transgenes were inherited in a Mendelian fashion.
Received: 15 December 1998 / Revision received: 2 July 1999 · Accepted: 8 July 1999 相似文献
16.
Somatic embryogenesis and plant regeneration in chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) 总被引:2,自引:0,他引:2
Somatic embryogenesis was observed in ray-floret explants of Dendranthema grandiflorum (Ramat.) Kitamura cv. Aboukyu on Murashige and Skoog medium containing high concentrations of 3-indoleacetic acid (IAA) and
kinetin. 1-Naphthaleneacetic acid also induced somatic embryogenesis but indole-3-butyric acid or 2,4-dichlorophenoxy acetic
acid did not. Other cytokinins, such as 6-benzylaminopurine (BAP) and thidiazuron, were also not effective. No embryos were
seen at lower IAA concentrations with kinetin and various concentrations of BAP, although higher BAP concentrations yielded
many adventitious shoots. In contrast, no somatic embryogenesis was observed from leaves using any combination of plant growth
regulators. Histologically, primordia showed a typical embryo shape with a well-developed vascular bundle between the shoot
and the root primordia. Embryos had both stomata cells and a root system with polarity. Plants were efficiently regenerated
from ray floret-derived embryos subcultured in the appropriate medium.
Received: 30 April 1999 / Revision received: 7 October 1999 / Accepted: 27 January 2000 相似文献
17.
T. Krugman A. Korol E. Nevo J. W. Snape O. Levy B. Rubin 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,94(1):46-51
Chlorotoluron is a selective phenylurea herbicide widely used for broad-leaved and annual grass weed control in cereals. Variation in the response to chlorotoluron (CT) was found in both hexaploid bread wheat (Triticum aestivum L.) and wild tetraploid wheat (Triticum dicoccoides KöRN.). Here, we describe the comparative mapping of the CT resistance gene (Su1) on chromosome 6B in bread and wild wheat using RFLP markers. In bread wheat, mapping was based on 58 F4 single-seed descent (SSD) plants of the cross between a genotype sensitive to chlorotoluron, ‘Chinese Spring’ (CS), and a resistant derivative, the single chromosome substitution line, CS (‘Cappele-Desprez’ 6B) [CS (CAP6B). In T dicoccoides, mapping was based on 37 F2 plants obtained from the cross between the CT-susceptible accession B-7 and the resistant accession B-35. Nine RFLP probes spanning the centromere were chosen for mapping. In bread wheat Su1 was found to be linked to α-Amy-1 (9.84 cM) and Xpsr371 (5.2 cM), both on the long arm of 6B, and Nor2 (2.74 cM) on the short arm. In wild wheat the most probable linkage map was Nor2-Xpsr312-Su1-Pgk2, and the genetic distances between the genes were 24.8cM, 5.3cM, and 6.8cM, respectively. These results along with other published map data indicate that the linear order of the genes is similar to that found in T. aestivum. The results of this study also show that the Su1 gene for differential response to chlorotoluron has evolved prior to the domestication of cultivated wheat and not in response to the development and use of chemicals. 相似文献
18.
Embryogenic tissue of Pinus patula Scheide et Deppe was cryopreserved for 8 weeks using sorbitol and dimethylsulfoxide (DMSO) as cryoprotectants. Results indicate
that 0.3 M sorbitol and 5% DMSO had the best cryoprotecting effect. The recovered tissue initially underwent a lag phase but then continued
to proliferate normally on MSG3 maintenance medium. Recovered tissue was placed onto MSG5 maturation medium, and embryos were
isolated and germinated. Plantlet regeneration from the recovered tissue was achieved.
Received: 16 April 1999 / Revision received: 26 July 1999 / Accepted: 17 August 1999 相似文献
19.
Manish Sainger Darshna Chaudhary Savita Dahiya Ranjana Jaiwal Pawan K. Jaiwal 《Physiology and Molecular Biology of Plants》2015,21(4):505-517
An efficient, rapid and direct multiple shoot regeneration system amenable to Agrobacterium-mediated transformation from primary leaf with intact petiole of blackgram (Vigna mungo) is established for the first time. The effect of the explant type and its age, type and concentration of cytokinin and auxin either alone or in combination and genotype on multiple shoot regeneration efficiency and frequency was optimized. The primary leaf explants with petiole excised from 4-day-old seedlings directly developed multiple shoots (an average of 10 shoots/ explant) from the cut ends of the petiole in 95 % of the cultures on MSB (MS salts and B5 vitamins) medium containing 1.0 μM 6-benzylaminopurine. Elongated (2–3 cm) shoots were rooted on MSB medium with 2.5 μM indole-butyric acid and resulted plantlets were hardened and established in soil, where they resumed growth and reached maturity with normal seed set. The regenerated plants were morphologically similar to seed-raised plants and required 8 weeks time from initiation of culture to establish them in soil. The regeneration competent cells present at the cut ends of petiole are fully exposed and are, thus, easily accessible to Agrobacterium, making this plant regeneration protocol amenable for the production of transgenic plants. The protocol was further successfully used to develop fertile transgenic plants of blackgram using Agrobacterium tumefaciens strain EHA 105 carrying a binary vector pCAMBIA2301 that contains a neomycin phosphotransferase gene (nptII) and a β-glucuronidase (GUS) gene (uidA) interrupted with an intron. The presence and integration of transgenes in putative T0 plants were confirmed by polymerase chain reaction (PCR) and Southern blot hybridization, respectively. The transgenes were inherited in Mendelian fashion in T1 progeny and a transformation frequency of 1.3 % was obtained. This protocol can be effectively used for transferring new traits in blackgram and other legumes for their quantitative and qualitative improvements. 相似文献
20.
Transformation and regeneration of garlic (Allium sativum L.) by Agrobacterium-mediated gene transfer 总被引:5,自引:0,他引:5
By using highly regenerative calluses, we developed a stable transformation system in garlic (Allium sativum L.). The temperature and number of days of co-cultivation with Agrobacterium tumefaciens was shown to be an important factor in transient expression of the uid A gene. After a culture period of 5 months in selection medium containing hygromycin, 20 shoots were induced from ca. 1000
calluses, among which 15 plants expressed β-glucuronidase activity upon staining with X-Gluc. Shoots developed into transgenic garlic after 1 month. Integration of the uid A gene was confirmed by Southern blot analysis for genomic DNA of transgenic garlic plants.
Received: 25 October 1999 / Revision received: 16 February 2000 / Accepted: 22 February 2000 相似文献