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1.
We have previously reported that expression of salt-responsive genes, including Bruguiera gymnorhiza
ankyrin repeat protein 1 (BgARP1), enhances salt tolerance in both Agrobacterium tumefaciens and Arabidopsis. In this report, we further characterized BgARP1-expressing Arabidopsis to elucidate the role of BgARP1 in salt tolerance. BgARP1-expressing plants exhibited more vigorous growth than wild-type plants on MS plates containing 125–175 mM NaCl. Real-time
PCR analysis showed enhanced induction of osmotin34 in the 2-week-old transformants under 125 mM NaCl. It was also showed that induction of typical salt-responsive genes, including
RD29A, RD29B, and RD22, was blunted and delayed in the 4-week-old transformants during 24 h after 200 mM NaCl treatment. Ion content analysis showed
that transgenic plants contained more K+, Ca2+, and NO3
−, and less NH4
+, than wild-type plants grown in 200 mM NaCl. Our results suggest that BgARP1-expressing plants may reduce salt stress by up-regulating osmotin34 gene expression and maintaining K+ homeostasis and regulating Ca2+ content. These results indicate that BgARP1 is functional on a heterogeneous background. 相似文献
2.
J. F. Liu C. Y. Zhao J. Ma G. Y. Zhang M. G. Li G. J. Yan X. F. Wang Z. Y. Ma 《Euphytica》2011,181(1):31-40
A phytase gene (phyA), isolated from Aspergillus ficuum (AF537344), was introduced into cotton (Gossypium hirsutum L.) by Agrobacterium-mediated transformation to increase the phosphorus (P) acquisition efficiency of cotton. Southern and Northern blot analyses
showed that the phyA was successfully incorporated into the cotton genome and expressed in transgenic lines. After growing for 45 days with phytate
(Po) as the only P source, the shoot and root dry weights of the transgenic plants all increased by nearly 2.0-fold relative
to those of wild-type plants, but were similar to those of transgenic plants supplied with inorganic phosphorus. The phytase
activities of root extracts prepared from transgenic plants were 2.4- to 3.6-fold higher than those from wild-type plants,
and the extracellular phytase activities of transgenic plants were also 4.2- to 6.3-fold higher. Furthermore, the expressed
phytase was secreted into the rhizospheres as demonstrated by enzyme activity staining. The transgenic plants accumulated
much higher contents of total P (up to 2.1-fold after 30 days of growth) than the wild-type plants when supplied with Po.
These findings clearly showed that cotton plant transformed with a fungal phytase gene was able to secret the enzyme from
the root, which markedly improved the plant’s ability to utilize P from phytate. This may serve as a promising step toward
the development of new cotton cultivars with improved phosphorus acquisition. 相似文献
3.
Takashi Nakatsuka Yoshiko Abe Yuko Kakizaki Akiko Kubota Norimoto Shimada Masahiro Nishihara 《Euphytica》2009,168(1):113-119
Ornamental gentian plants are perennial and have a juvenile period of over 1 year before flowering. We transformed gentian
plants with a construct comprising the Arabidopsis
FLOWERING LOCUS T (FT) gene (encoding a major component protein of the flowering hormone ‘florigen’) under the control of the rolC promoter from Agrobacterium rhizogenes, which is known to induce vascular-specific expression. The resultant rolCpro-FT transgenic gentian plants showed early flowering in vitro and the earliest line formed floral buds within 4 months
after transformation. Regeneration experiments from leaf explants of these rolCpro-FT transgenic plants also confirmed the early flowering phenotype. After acclimatization, these transgenic plants showed
normal floral development in a closed greenhouse. There is no effective method to induce early flowering by cultivation management
in gentian, therefore these lines might be very useful as annual early season cultivars. 相似文献
4.
5.
Homozygous transgenic cotton (Gossypium hirsutum L.) plants that accumulated glycinebetaine (GB) in larger quantities were more tolerant to salt than wild-type (WT) plants.
Four transgenic lines, namely 1, 3, 4, and 5, accumulated significantly higher levels of GB than WT plants did both before
and after salt stress. At 175 and 275 mM NaCl, seeds of all the transgenic lines germinated earlier and recorded a higher
final germination percentage, and the seedlings grew better, than those of the WT. Under salt stress, all the lines showed
some characteristic features of salt tolerance, such as higher leaf relative water content (RWC), higher photosynthesis, better
osmotic adjustment (OA), lower percentage of ion leakage, and lower peroxidation of the lipid membrane. Levels of endogenous
GB in the transgenic plants were positively correlated with RWC and OA. The results indicate that GB in transgenic cotton
plants not only maintains the integrity of cell membranes but also alleviates osmotic stress caused by high salinity. Lastly,
the seed cotton yield of transgenic lines 4 and 5 was significantly higher than that of WT plants in saline soil. This research
indicates that betA gene has the potential to improve crop’s salt tolerance in areas where salinity is limiting factors for agricultural productivity. 相似文献
6.
P. T. Lynch J. Jones N. W. Blackhall M. R. Davey J. B. Power E. C. Cocking M. R. Nelson D. M. Bigelow T. V. Orum C. E. Orth W. Schuh 《Euphytica》1955,85(1-3):395-401
Summary The phenotypes of seed progeny (R2 generation) of Oryza sativa L. cv. Taipei 309, which carried the neomycin phosphotransferase II (npt II) gene, were compared with those of non-transformed, protoplast-derived plants of the same generation and non-transformed, seed-derived plants under field and glasshouse conditions. Under both conditions the transgenic plants were generally smaller, took longer to flower and had reduced fertility. Significant differences were observed between individuals within the group of transgenic plants. The npt II gene was present in most of the transgenic plants, but NPT II activity was only detected in a minority of individuals. 相似文献
7.
The performance of insect-resistant transgenic poplar trees (Populusnigra) expressing a Cry1Ac gene from Bacillus thuringiensis subsp. Kurstaki HD-1 against poplar defoliators was evaluated in the field at the Manas Forest Station in Xinjiang Uygur
Autonomous Region during1994–1997. The results showed that the average percentage of highly damaged leaves on the transgenic
trees was 10% while that on the control trees in nearby plantations reached 80–90%. The average number of pupae per m2 of soil at 20cm depth in transgenic poplar plantation was 18 which was only 20% of that found in the non-transgenic control
field. The number of pupae and the leaf-damage on transgenic trees described above are all far below the threshold set for
chemical protection measures. The non-transformed poplar trees grown in the same plantation with the transgenic trees were
also protected indicating that cross protection occurred between these two kinds of trees. Insect-resistant transgenic poplar
trees have a potential application value in afforestation.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
8.
Xuyin Qin Yang Liu Shanjing Mao Tianbi Li Hsinkan Wu Chengcai Chu Youping Wang 《Euphytica》2011,177(1):33-43
Embryogenic callus of Phalaenopsis amabilis derived from leaf tissue was cocultivated with Agrobacterium
tumefaciens strain LBA4404 harboring a plant cloning vector. The vector carried the lipid transfer protein (LTP) encoding gene cloned
from cold tolerant Brazilian upland rice cv. IAPAR 9. The highest transformation efficiency (12.16%) was obtained when 1–2 mm
calli were infected and cocultivated with 0.4 (OD600) A. tumefaciens for 20 min. Transgene integration of kan-resistant plants was confirmed through polymerase chain reaction analysis and Southern hybridization. Four hundred seventy
transgenic plants, each derived from an independent protocorm-like body, were obtained. The expression of rice cold-inducible
LTP gene in transgenic P. amabilis improved its adaptive responses to cold stress. The examination of transgenic plants revealed that enhanced cold tolerance
was most likely due to the increased accumulation of several compatible solutes such as total soluble sugars, proline, antioxidant
superoxide dismutase, decreased accumulation of malondialdehyde, and maintained electrolytes within the membrane compared
with controls. 相似文献
9.
Summary Semilooper resistant transgenic castor plants were produced through Agrobacterium-mediated genetic transformation method. Two castor cultivars, Jyothi and VP1 were transformed using the super-binary vector
pTOK233 carrying gus A and hpt genes. Putative transformants were regenerated following selection on the hygromycin containing medium. GUS positive primary
transformants, when subjected to Southern analysis, revealed stable integration of gus A into their genomes. In the T1 generation, a monogenic segregation ratio of 3 GUS positive: 1 GUS negative plants was observed. Furthermore, transformation
experiments were carried out with the Agrobacterium pSB111 super-binary vector carrying a synthetic delta endotoxin gene cryIAb and the herbicide resistance gene bar both driven by cauliflower mosaic virus 35S promoter. Putative transformants were regenerated through selection on the phosphinothricin
containing medium and Basta tolerant transformants were subjected to molecular analysis. PCR analysis revealed the presence
of both bar and cryIAb genes in the Basta tolerant primary transformants. Southern analysis of PCR positive plants with cryIAb probe showed a 3 Kb band upon HindIII digestion and a > 6 Kb band with BamHI digestion, thus suggesting stable integration of cryIAb intact expression cassette and independent nature of the transformants. The primary transformants subjected to ELISA disclosed
varied levels of Cry protein. These transgenics expressing cryIAb – when bioassayed against freshly hatched semilooper larvae – induced substantial (> 88%) insect mortality. Southern analysis
of 2T1 plants revealed the presence of cryIAb gene, indicating stable inheritance of the transgene into the next generation. In T1, all the Southern-positive plants for cryIAb invariably exhibited tolerance to Basta, denoting co-segregation of both bar and cryIAb genes. Transgenics, expressing cryIAb exhibited ample resistance against the castor semilooper. 相似文献
10.
The variation obtained in storage fatty acids induced by the procedures of tissue culture and transformation with Agrobacterium tumefaciens was investigated and compared in rapeseed, Brassica napus, cv. Hanna. An increased variation in the fatty acid profiles was noted after tissue culture and transformation compared
with plants derived directly from seeds. In the second generation of rapeseed transformants, T2, the content of oleic acid ranged from 39–72%, 12–31% for linoleic acid and 7–16% for linolenic acid. This could be compared
with the oleic acid content in the T2 generation of tissue culture-derived plants which ranged between 47–76% and in seed-derived material where oleic acid ranged
between 55–69%.In the T3 generation the ranges in transgenic seeds were decreased but still larger than in the seed derived plants. The range in transgenic
plants was 49–64% for oleic acid, 20–28% for linoleic acid and 9–18% for linolenic acid. The most extreme individuals, both
highest and lowest in the common fatty acids, were found in the group of transformed plants independent of generation. The
total lipid content was also affected by the two treatments and seeds with the lowest and highest lipid content were both
found among the transformed plants. In conclusion, care should be taken to use proper controls when performing transformation
experiments in order to distinguish variation in the fatty acid profiles induced by the transformation procedure and tissue
culture treatments from the changes due to transgenic expression.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
11.
Arthikala Manoj Kumar Rohini Sreevathsa Kalpana Nanja Reddy Prasa Trichy Ganesh Makarla Udayakumar 《Journal of Crop Science and Biotechnology》2011,14(2):125-132
Agrobacterium tumefaciens mediated in planta transformation protocol was developed for castor, Ricinus communis. Two-day-old seedlings were infected with Agrobacterium strain EHA105/pBinBt8 harboring cry1AcF and established in the greenhouse. Screening the T1 generation seedlings on 300 mg L−1 kanamycin identified the putative transformants. Molecular and expression analysis confirmed the transgenic nature and identified
high-expressing plants. Western blot analysis confirmed the co-integration of the nptII gene in the selected transgenic plants. Bioassay against Spodoptera litura corroborated with high expression and identified five promising effective lines. Analysis of the T2 generation plants proved the stability of the transgene indicating the feasibility of the method. 相似文献
12.
13.
Random amplified polymorphic DNA analysis of the moth orchids, Phalaenopsis (Epidendroideae: Orchidaceae) 总被引:1,自引:0,他引:1
The inheritance and stability of the acetolactate synthase (als) transgene were compared in transgenic maize plants, generated using the pollen-tube pathway, particle bombardment, or Agrobacterium-mediated methods of transformation. Progeny populations generated by successive selfing or backcrossing of primary transformants
were analyzed over three generations, using PCR and herbicide screening, to examine segregation and als activity, respectively, and transgenic homozygous plants were selected. The pollen-tube method resulted in a higher rate
of primary normal transgenic plants and a less-stable transmission of the als locus than did the other two methods. When transferred by the particle bombardment and Agrobacterium-mediated methods, the als gene was in a much higher proportion of Mendelian transmission than transferred by the pollen-tube method. Compared to the
Agrobacterium-mediated transformation, the particle bombardment method tends to create multiple copies and insert sites of the als gene in maize genome, which delaying the homogenization of the als locus with advancing generations. Agrobacterium-mediated transformation resulted in a greater proportion of stable, low copy number (in general 1–2) transgenic events, facilitating
the stable inheritance of the als gene, and producing multiple desirable transgenic plants. 相似文献
14.
Tetraploid plants were induced successfully from diploid bananas Musa acuminata (AA genome) ‘Kluai Leb Mu Nang’ and ‘Kluai Sa’ (2n = 2x = 22) with in vitro oryzalin treatment. Calluses from in vitro-grown shoot tips of both cultivars were treated with oryzalin
at concentrations of 1.5 or 3 mg l−1 for 24, 48 and 72 h, respectively. The oryzalin treatments produced tetraploids at a frequency of 15.6% in ‘Kluai Leb Mu
Nang’ and 16.7% in ‘Kluai Sa’ as detected by flow cytometry. Chromosome counting showed that the tetraploid plant chromosome
number was (2n = 4x = 44). The selected tetraploid plants were transplanted in the field and variations in the morphological characteristic of
leaf shape and fruit bunch compared to normal diploid plants were found under the same growing condition even after 3 years
of cultivation. 相似文献
15.
Morad Jafari Peyman Norouzi Mohammad Ali Malboobi Behzad Ghareyazie Mostafa Valizadeh Seyed Aboulghasem Mohammadi Mozhgan Mousavi 《Euphytica》2009,165(2):333-344
Two diploid sugar beet genotypes of agronomical importance were transformed using Agrobactrium tumefaciens harboring pBI35Scry containing a synthetic cry1Ab gene. Leaf blade with attached shoot bases, a highly regenerative tissue, were used as explant substratum for transformation.
PCR screening with cry1Ab-specific primers showed the presence of transgene in more than 50% of the regenerated kanamycin-resistant plants after treatment
with the antibiotic. A transformation rate of 8.8–12.2% (depending on genotype) was achieved as revealed by genomic DNA dot
blotting. The intact integration of transgene cassette into the genome was furthermore confirmed by Southern blot analysis.
The expression of the cry1Ab gene encoding a truncated endotoxin (67 kDa) at about 0.1% of total soluble protein was achieved in the leaves of transgenic
plants as shown by Western blot analysis. Bioassays under in vitro conditions with Spodoptera littoralis, one of the most important pests in sugar beet fields, demonstrated enhanced resistance against this pest. The inheritance
of the inserted transgene was confirmed in F1 plants obtained through crossing of T0 plants with a cytoplasmic male sterile line. Transgenic plants are currently grown in a greenhouse and will be subjected
to further bioassay analyses against other lepidopteran pests of sugar beet. 相似文献
16.
Junjie Zhang Fan Liu Lei Yao Chen Luo Yue Yin Guixiang Wang Yubi Huang 《Breeding Science》2012,62(2):105-112
Lepidopteran larvae are the most injurious pests of Chinese cabbage production. We attempted the development of transgenic Chinese cabbage expressing the potato proteinase inhibitor II gene (pinII) and bioassayed the pest-repelling ability of these transgenic plants. Cotyledons with petioles from aseptic seedlings were used as explants for Agrobacterium-mediated in vitro transformation. Agrobacterium tumefaciens C58 contained the binary vector pBBBasta-pinII-bar comprising pinII and bar genes. Plants showing vigorous PPT resistance were obtained by a series concentration selection for PPT resistance and subsequent regeneration of leaf explants dissected from the putative chimera. Transgenic plants were confirmed by PCR and genomic Southern blotting, which showed that the bar and pinII genes were integrated into the plant genome. Double haploid homozygous transgenic plants were obtained by microspore culture. The pinII expression was detected using quantitative real time polymerase chain reaction (qRT-PCR) and detection of PINII protein content in the transgenic homozygous lines. Insect-feeding trials using the larvae of cabbage worm (Pieris rapae) and the larvae of the diamondback moth (Plutella xylostella) showed higher larval mortality, stunted larval development, and lower pupal weights, pupation rates, and eclosion rates in most of the transgenic lines in comparison with the corresponding values in the non-transformed wild-type line. 相似文献
17.
Bushra Rashid Ghazanfar Ali Khan Tayyab Husnain Sheikh Riazuddin 《Journal of Crop Science and Biotechnology》2009,12(3):135-141
We report the field evaluation of second generation of transgenic cotton expressing Bacillus thuringiensis (Bt) genes cry1Ac and cry2A under CaMV 35S promoter. Sixty-five transgenic lines were grown under RCBD design. Transgenic plants exhibited inherent ability to resist
target insect (p < 0.05 and 0.01). Morphological studies showed significant reduction in plant height making them favorable
for breeding. Yield was significantly increased for the transgenic lines. Fiber analysis showed improved gin turn out 40%
for transgenic lines in comparison to 32% for non-transformed lines. Fibre quality of transgenic lines was not affected when
compared with non transgenic lines. Inheritance pattern for transgenic lines suggests the need of further studies to understand
the complex molecular mechanisms for resistance management and biosafety studies to develop new Bt cotton varieties. 相似文献
18.
19.
André M. Almeida Enrique Villalobos Susana S. Araújo Barbara Leyman Patrick Van Dijck Luís Alfaro-Cardoso Pedro S. Fevereiro José M. Torné Dulce M. Santos 《Euphytica》2005,146(1-2):165-176
Summary Trehalose (a non-reducing disaccharide) plays an important role in abiotic stress protection. It has been shown that using
trehalose synthesis genes of bacterial origin, drought and salt tolerance could be achieved in several plants. A cassette
harboring the AtTPS1 gene under the control of the CaMV35S promoter and the Bialaphos resistance gene was inserted in the binary plasmid vector
pGreen0229 and used for Agrobacterium-mediated transformation of tobacco (Nicotiana tabacum). T0 plants obtained were analyzed by PCR for the presence of AtTPS1 gene. Thirty lines were positive and seeds were germinated on media with 6 mg/l PPT to obtain T1 plants that were grown in
the greenhouse to obtain T2 seeds that were germinated on selective media. Lines which seeds showed a 100 % survival rate
were considered homozygous transgenic T1 lines. Three lines were selected and gene expression confirmed by northern and western
blots. Transgenic seeds were germinated on media with different concentrations of mannitol (0, 0.25, 0.5 and 0.75 M) and sodium
chloride (0, 0.07, 0.14, 0.2, 0.27 and 0.34 M) to score their tolerance to osmotic stress. Assays were conducted to test the
tolerance of transgenic plants to drought (measurement of water percentage as a consequence of water withdrawal), desiccation
(measurement of water loss as a consequence leaf detaching) and temperature stresses (germination at 15 ∘C and 35∘C). Transgenic tobacco plant lines registered higher germination rates under osmotic and temperature stress situations than
did wild-type plants. Responses to drought and desiccation stresses were similar for all plant lines. It can hence be suggested
that the heterologous expression of TPS1 gene from Arabidopsis can be used successfully to increase abiotic stress tolerance in model plants and probably in other crops. 相似文献
20.
Zhongxia Luo Xiongjian Zhang Jingyi Chen Zhangying Wang Lifei Huang Hongda Zou Zhufang Yao Yiling Yang Boping Fang 《Plant Breeding》2020,139(2):381-388
The genetic behaviour of a yellow-green leaf sweetpotato mutant, a269, was analysed to prepare for genetic mapping and cloning of this mutant and to elucidate the regulation of its molecular mechanisms. The phenotypic stability of the mutant was observed under natural conditions, and photosynthetic pigments in mature leaves were measured. GN1323 and a269 were crossed and backcrossed. Genetic behaviour was assessed using phenotypic expression and segregation in F1, BC1 and BC2. The a269 leaves were yellow-green at all stages after germination. The thylakoid lamellae of a269 were disorganized, had incomplete grana and were fewer compared with GN1323. The direct and reciprocal F1 populations had normal leaf colour, whereas the BC2 populations had yellow-green leaves. Segregation of plants with normal- and yellow-green leaf colour in the four BC1 populations satisfied the backcross segregation ratio of 11 : 1 for auto-allohexaploids. The a269 mutant is stable, and its trait is controlled by one recessive nuclear locus. 相似文献