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1.
Lin WC Lu CF Wu JW Cheng ML Lin YM Yang NS Black L Green SK Wang JF Cheng CP 《Transgenic research》2004,13(6):567-581
Development of effective disease-resistance to a broad-range of pathogens in crops usually requires tremendous resources and effort when traditional breeding approaches are taken. Genetic engineering of disease-resistance in crops has become popular and valuable in terms of cost and efficacy. Due to long-lasting and broad-spectrum of effectiveness against pathogens, employment of systemic acquired resistance (SAR) for the genetic engineering of crop disease-resistance is of particular interest. In this report, we explored the potential of using SAR-related genes for the genetic engineering of enhanced resistance to multiple diseases in tomato. The Arabidopsis NPR1 (nonexpresser of PR genes) gene was introduced into a tomato cultivar, which possesses heat-tolerance and resistance to tomato mosaic virus (ToMV). The transgenic lines expressing NPR1 were normal as regards overall morphology and horticultural traits for at least four generations. Disease screens against eight important tropical diseases revealed that, in addition to the innate ToMV-resistance, the tested transgenic lines conferred significant level of enhanced resistance to bacterial wilt (BW) and Fusarium wilt (FW), and moderate degree of enhanced resistance to gray leaf spot (GLS) and bacterial spot (BS). Transgenic lines that accumulated higher levels of NPR1 proteins exhibited higher levels and a broader spectrum of enhanced resistance to the diseases, and enhanced disease-resistance was stably inherited. The spectrum and degree of these NPR1-transgenic lines are more significant compared to that of transgenic tomatoes reported to date. These transgenic lines may be further explored as future tomato stocks, aiming at building up resistance to a broader spectrum of diseases. 相似文献
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Kim MJ Federer W Mutschler MA 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2005,112(1):21-29
The need for a new analytical approach was encountered in the course of characterizing newly developed tomato lines resistant to late blight. Late blight resistant tomato lines were created in independent breeding programs using the accession Solanum pimpinellifolium L. (formerly Lycopersicon pimpinellifolium (L.) Miller) L3708 as the source of the resistance. However, initial field observation suggested that the late blight resistance in the lines produced by two independent breeding programs differed. Possible causes included a partial transfer of the late blight resistance derived from S. pimpinellifolium L3708 or the possibility of race specificity of this resistance. A crucial issue was determining the most appropriate and robust analytical method to use with data from laboratory analyses of the responses of nine tomato lines against five P. infestans isolates. Prior analysis by standard ANOVA revealed significant differences across tomato lines but could not determine whether the disease responses in the CLN-R lines were different from those of the heterozygous F1 hybrids, created by crossing susceptible tomatoes with the fixed CU-R lines. A different analytical method was needed. Therefore, sporangia numbers/leaflet and diseased area data were analyzed using a half-normal probability plot and regression analysis. The results of this analysis show its utility for genetic or pathology studies. Considering only populations of the uniform tomato lines, this method confirms the results obtained by using a standard ANOVA, but provides a clearer demonstration of the distributions of the individuals within the populations and how this distribution impacts variance and the difference among the populations. This method also allows a joint analysis of the uniform lines with an additional population that is less uniform, because it is segregating. Such an analysis would be invalid using a standard ANOVA. The results of this joint analysis determined that the additional population was divergent from the fixed CU-R lines, and, against some isolates, against the CLN-R lines as well. Half-normal probability plot analysis method would be applicable more broadly beyond analysis of disease resistance data. It could be useful for data from populations that are not normally distributed, for traits which are affected by epistatic gene action, and could be useful for selection of extremes. 相似文献
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Overexpression of defense response genes in transgenic wheat enhances resistance to Fusarium head blight 总被引:2,自引:0,他引:2
Mackintosh CA Lewis J Radmer LE Shin S Heinen SJ Smith LA Wyckoff MN Dill-Macky R Evans CK Kravchenko S Baldridge GD Zeyen RJ Muehlbauer GJ 《Plant cell reports》2007,26(4):479-488
Fusarium head blight (FHB) of wheat, caused by Fusarium graminearum and other Fusarium species, is a major disease problem for wheat production worldwide. To combat this problem, large-scale
breeding efforts have been established. Although progress has been made through standard breeding approaches, the level of
resistance attained is insufficient to withstand epidemic conditions. Genetic engineering provides an alternative approach
to enhance the level of resistance. Many defense response genes are induced in wheat during F. graminearum infection and may play a role in reducing FHB. The objectives of this study were (1) to develop transgenic wheat overexpressing
the defense response genes α-1-purothionin, thaumatin-like protein 1 (tlp-1), and β-1,3-glucanase; and (2) to test the resultant
transgenic wheat lines against F. graminearum infection under greenhouse and field conditions. Using the wheat cultivar Bobwhite, we developed one, two, and four lines
carrying the α-1-purothionin, tlp-1, and β-1,3-glucanase transgenes, respectively, that had statistically significant reductions
in FHB severity in greenhouse evaluations. We tested these seven transgenic lines under field conditions for percent FHB disease
severity, deoxynivalenol (DON) mycotoxin accumulation, and percent visually scabby kernels (VSK). Six of the seven lines differed
from the nontransgenic parental Bobwhite line for at least one of the disease traits. A β-1,3-glucanase transgenic line had
enhanced resistance, showing lower FHB severity, DON concentration, and percent VSK compared to Bobwhite. Taken together,
the results showed that overexpression of defense response genes in wheat could enhance the FHB resistance in both greenhouse
and field conditions. 相似文献
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Arabidopsis LSD1-related proteins that contain LSD1-like zinc finger domains have been identified to be involved in disease resistance
and programmed cell death. To investigate the potential role of LSD1-related gene in rice (Oryza sativa L.), we cloned an LSD1 ortholog, OsLOL2, from the rice cDNA plasmid library. The OsLOL2 gene is predicted to encode a polypeptide of 163 amino acids with two LSD1-like zinc finger domains with 74.5% identity to
those of LSD1. Southern blot analysis indicated that OsLOL2 was a single-copy gene in the rice genome. Transgenic rice lines carrying the antisense strand of OsLOL2 with decreased expression of OsLOL2 had dwarf phenotypes, and the dwarfism could be restored by exogenous GA3 treatment, suggesting that the dwarfism was the result of a deficiency in bioactive gibberellin (GA). In agreement with this
possibility, the content of endogenous bioactive GA1 decreased in the antisense transgenic lines. Expression of OsKS1, one of the genes encoding for GA biosynthetic enzymes, was suppressed in the antisense transgenic lines. Sense transgenic
lines with increased expression of OsLOL2 were more resistant to rice bacterial blight, while antisense transgenic lines were less resistant to rice bacterial blight.
The OsLOL2-GFP (green fluorescence protein) fusion protein was localized in the nucleus of cells of transgenic BY2 tobacco
(Nicotiana tabacum L.). These data suggest that OsLOL2 is involved in rice growth and disease resistance. 相似文献
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Several root-knot nematode (Meloidogyne spp.) resistance genes have been discovered in different pepper (Capsium annuum L.) lines; however, none of them has yet been cloned. In this study, a candidate root-knot nematode resistance gene (designated
as CaMi) was isolated from the resistant pepper line PR 205 by degenerate PCR amplification combined with the RACE technique. Expression
profiling analysis revealed that this gene was highly expressed in roots, leaves, and flowers and expressed at a lower level
in stems and was not detectable in fruits. To verify the function of CaMi, a sense vector containing the genomic DNA spanning the full coding region of CaMi was constructed and transferred into root-knot nematode susceptible tomato plants. Sixteen transgenic plants carrying one
to five copies of T-DNA inserts were generated from two nematode susceptible tomato cultivars. RT-PCR analysis revealed that
the expression levels of CaMi gene varied in different transgenic plants. Nematode assays showed that the resistance to root-knot nematodes was significantly
improved in some transgenic lines compared to untransformed susceptible plants, and that the resistance was inheritable. Ultrastructure
analysis showed that nematodes led to the formation of galls or root knots in the susceptible lines while in the resistant
transgenic plants, the CaMi gene triggered a hypersensitive response (HR) as well as many necrotic cells around nematodes.
Rugang Chen and Hanxia Li are contributed equally to this work. 相似文献
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Mj-AMP2, a knottin-type antimicrobial peptide, in vitro inhibits the growth of several plant pathogenic fungi including Magnaporthe oryzae. We demonstrate that transgenic rice (Oryza sativa L.) plants expressing the Mj-AMP2 gene show enhanced resistance to M. grisea, the causal agent of the rice blast disease. Mj-AMP2 was efficiently expressed and the level of Mj-AMP2 ranged from 0.32% to 0.38% of the total protein in the transgenic rice plants. In vitro inhibitory activity assays with the crude protein extract from transgenic rice indicated that the Mj-AMP2 protein produced was biologically active. Constitutive expression of Mj-AMP2 in transgenic rice reduces the growth of M. grisea by 63% with respect to untransformed control plant, and no effect on plant phenotype was observed. Transgene expression of Mj-AMP2 gene was not accompanied by an induction of pathogenesis-related (PR) gene expression indicating that the transgene product itself is directly active against the pathogen. The results presented in this study suggest that the Mj-AMP2 gene could be a useful candidate for protection of rice plants against the rice blast fungus M. grisea. 相似文献
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Gubba Augustine Gonsalves Carol Stevens Mikel R. Tricoli David M. Gonsalves Dennis 《Molecular breeding : new strategies in plant improvement》2002,9(1):13-23
This study was undertaken to develop tomato plants with broad resistanceto tospoviruses which are a major limiting factor to tomato productionworldwide. A nontransgenic tomato line Stevens-Rodale (S-R), six transgenictomato lines expressing the nucleocapsid (N) protein gene of the lettuceisolate of tomato spotted wilt virus (TSWV-BL), and progeny of the crosses between S-Rand three of the transgenic lines homozygous for the N gene were evaluated fortheir resistance to tospovirus infection in greenhouse inoculation tests. S-Rhas the Sw-5 gene that confers resistance to several TSWVisolates. The six transgenic lines showed high levels of resistance wheninoculated with either TSWV-BL or a tomato isolate from Hawaii (TSWV-H).However, these same plants were highly susceptible to the Brazilian isolate ofgroundnut ringspot virus (GRSV-BR). Plants with the Sw-5gene were resistant to TSWV-BL and GRSV-BR, but were susceptible to TSWV-H.When inoculated with any of the three viruses, the F1 progeny of thecrosses exhibited a susceptible, tolerant, or resistant phenotype with a higherproportion of the plants being either tolerant or resistant. When F2progeny from F1 resistant plants of each cross were inoculated withany of the three viruses, a higher proportion of tolerant and resistant plantswas observed compared to the F1 progeny. Our results show thepotential to obtain broad resistance to tospoviruses by combining transgenicand natural resistance in a single plant. 相似文献
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Transgenic tomato resistant to tomato leaf curl disease (ToLCD) using replicase (rep) gene sequences of Tomato leaf curl virus in antisense orientation were developed via Agrobacterium-mediated transformation. A binary vector carrying the antisense rep gene (untranslatable full length sequence, 1086 bp) along with the npt II gene was used for transformation. High level of resistance and inheritability of the transgene was observed up to T2 stage following challenge inoculation with the virus. The mechanism of resistance appears RNA-mediated, since the plants carried the untranslatable antisense rep gene. Progeny analysis of these plants showed classical Mendelian pattern of inheritance in two of the six transgenic lines having single transgene insertion. 相似文献
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Raham Sher Khan Rinaldi Sjahril Ikuo Nakamura Masahiro Mii 《Plant biotechnology reports》2008,2(1):13-20
Potato (Solanum tuberosum L.), one of the most important food crops, is susceptible to a number of devastating fungal pathogens in addition to bacterial
and other pathogens. Producing disease-resistant cultivars has been an effective and useful strategy to combat the attack
of pathogens. Potato was transformed with Agrobacterium tumefaciens strain EHA101 harboring chitinase, (ChiC) isolated from Streptomyces griseus strain HUT 6037 and bialaphos resistance (bar) genes in a binary plasmid vector, pEKH1. Polymerase chain reaction (PCR) analysis revealed that the ChiC and bar genes are integrated into the genome of transgenic plants. Different insertion sites of the transgenes (one to six sites
for ChiC and three to seven for bar) were indicated by Southern blot analysis of genomic DNA from the transgenic plants. Expression of the ChiC gene at the messenger RNA (mRNA) level was confirmed by Northern blot analysis and that of the bar gene by herbicide resistance assay. The results obviously confirmed that the ChiC and bar genes are successfully integrated and expressed into the genome, resulting in the production of bialaphos-resistant transgenic
plants. Disease-resistance assay of the in vitro and greenhouse-grown transgenic plants demonstrated enhanced resistance against
the fungal pathogen Alternaria solani (causal agent of early blight). 相似文献
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Expression of a synthetic cry1EC gene for resistance against Spodoptera litura in transgenic peanut (Arachis hypogaea L.) 总被引:1,自引:1,他引:0
The tobacco cutworm (Spodoptera litura) is a polyphagous foliage insect and a major pest on peanut (Arachis hypogaea L.). S. litura is susceptible to the chimeric delta-endotoxin Cry1EC reported earlier. De-embryonated cotyledon explants of peanut were transformed using Agrobacterium tumefaciens strain EHA101 harboring a synthetic cry1EC gene driven by the CaMV 35S promoter. Transgenic plants of peanut with a single copy insertion of cry1EC were selected in the T(0) generation by Southern blot hybridization. Real-time PCR, Western blot and ELISA analysis indicated that expression of the cry1EC gene was higher in single copy T(1) plants. Immunoassay showed expression of Cry1EC up to 0.13% of total soluble protein in T(1) plants. Leaf feeding bioassay on highly expressing transgenic lines showed 100% killing of larvae at the 2(nd) instar stage of S. litura. This is the first report of transgenic peanut plants with resistance to S. litura. 相似文献
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Summary We present a strategy for establishing a transgenic doubled haploid maize line from heterozygous transgenic material by means
of anther culture. Compared to conventional inbreeding, the in vitro androgenesis technique enables a faster generation of virtually fully homozygous lines. Since the androgenic response is
highly genotype-dependent, we crossed transgenic, non-androgenic plants carrying a herbicide resistance marker gene (pat, encoding for phosphinothricin acetyl transferase) with a highly androgenic genotype. The transgenic progenies were used
as donor plants for anther culture. One transgenic and three non-transgenic doubled haploid lines have been established within
approximately 1 yr. The homozygosity of all four doubled haploid lines was tested by analysis of simple sequence repeat (SSR)
markers at 19 different loci. Polymorphisms were found between the lines but not within the lines indicating the homozygous
nature of the entire plant genome gained by anther culture. Southern blot analysis revealed that the transgenic donor plants
and their doubled haploid progeny exhibited the same integration pattern of the pat gene. No segregation of the herbicide resistance trait has been observed among the progeny of the transgenic doubled haploid
line. 相似文献
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The Sweet Pepper Ferredoxin-Like Protein (pflp) Conferred Resistance Against Soft Rot Disease in Oncidium Orchid 总被引:3,自引:0,他引:3
Liau CH Lu JC Prasad V Hsiao HH You SJ Lee JT Yang NS Huang HE Feng TY Chen WH Chan MT 《Transgenic research》2003,12(3):329-336
Genetic engineering to date has not been used to introduce disease resistance genes into the orchid gene pool. The ferredoxin-like protein gene originally isolated from sweet pepper is thought to function as a natural defense against infection due to its antimicrobial properties. Hence it was reasoned that introduction of this gene might produce Oncidium plants resistant to Erwinia carotovora, the causal agent for the soft rot disease. An expression vector containing sweet pepper ferredoxin-like protein (pflp) cDNA, hph and gusA coding sequence was successfully transformed into protocorm-like bodies (PLBs) of Oncidium orchid, using Agrobacterium tumefaciens strain EHA105. A total of 17 independent transgenic orchid lines was obtained, out of which six transgenic lines (-glucuronidase (GUS) positive) were randomly selected and confirmed by Southern, northern and western blot analyses. A bioassay was conducted on the transgenic lines. Transgenic plants showed enhanced resistance to E. carotovora, even when the entire plant was challenged with the pathogen. Our results suggest that pflp may be an extremely useful gene for genetic engineering strategies in orchids to confer resistance against soft rot disease. 相似文献