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Genetic transformation of strawberry: Stable integration of a gene to control biosynthesis of ethylene 总被引:7,自引:0,他引:7
Helena Mathews W. Wagoner J. Kellogg R. Bestwick 《In vitro cellular & developmental biology. Plant》1995,31(1):36-43
Summary Efficient methods ofAgrobacterium-mediated transformation are described for two Pacific Northwest cultivars of strawberry (Fragaria ×ananassa), Tristar and Totem. We report stable incorporation of a gene for control of ethylene biosynthesis, into strawberry (cultivar
Totem) for the first time. Cultivar Tristar was transformed with disarmed strains ofAgrobacterium tumefaciens (A. tumefaciens), LBA4404 or EHA101, containing a binary vector with marker genesuidA andnptII. Cultivar Totem was transformed withA. tumefaciens strains EHA101 or EHA105 harboring binary vectors with selectable marker genesnptII orhpt and with a gene for S-adenosylmethionine hydrolase (SAMase) for control of ethylene biosynthesis. The frequency of transgenic
shoots ranged from 12.5% to 58.8% of the original treated explants when using plasmids containing the gene encoding SAMase.
Primary shoot regenerants obtained on selection medium were subjected to several iterations of tissue isolation and reculture
on higher stringency selection medium for recovering uniformly transformed plantlets. Transgenic plants were confirmed by
their ability to undergo rooting on medium with selection at 60 mg/liter kanamycin or 10 mg/liter hygromycin. About 95–100%
of the transformation events from different experiments were capable of profuse rooting in the presence of selection. Insertion
of the SAMase gene and its integration into the strawberry genome were confirmed by Southern hybridization. About 500 plants
from 250 independent transgenic events have been successfully transferred to soil for further evaluation. 相似文献
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Xiaohua Qi Qianqian Li Xiaotian Ma Chunlu Qian Huihui Wang Nannan Ren Chenxi Shen Shumiao Huang Xuewen Xu Qiang Xu Xuehao Chen 《Plant, cell & environment》2019,42(5):1458-1470
Development of adventitious roots (ARs) at the base of the shoot is an important adaptation of plants to waterlogging stress; however, its physiological mechanisms remain unclear. Here, we investigated the regulation of AR formation under waterlogged conditions by hormones and reactive oxygen species (ROS) in Cucumis sativus L., an agriculturally and economically important crop in China. We found that ethylene, auxin, and ROS accumulated in the waterlogged cucumber plants. On the other hand, application of the ethylene receptor inhibitor 1‐methylcyclopropene (1‐MCP), the auxin transport inhibitor 1‐naphthylphthalamic acid (NPA), or the NADPH oxidase inhibitor diphenyleneiodonium (DPI) decreased the number of ARs induced by waterlogging. Auxin enhanced the expression of ethylene biosynthesis genes, which led to ethylene entrapment in waterlogged plants. Both ethylene and auxin induced the generation of ROS. Auxin‐induced AR formation was inhibited by 1‐MCP, although ethylene‐induced AR formation was not inhibited by NPA. Both ethylene‐ and auxin‐induced AR formation were counteracted by DPI. These results indicate that auxin‐induced AR formation is dependent on ethylene, whereas ethylene‐induced AR formation is independent of auxin. They also show that ROS signals mediate both ethylene‐ and auxin‐induced AR formation in cucumber plants. 相似文献
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Activation of ethylene signaling pathways enhances disease resistance by regulating ROS and phytoalexin production in rice
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Chao Yang Wen Li Jidong Cao Fanwei Meng Yongqi Yu Junkai Huang Lan Jiang Muxing Liu Zhengguang Zhang Xuewei Chen Koji Miyamoto Hisakazu Yamane Jinsong Zhang Shouyi Chen Jun Liu 《The Plant journal : for cell and molecular biology》2017,89(2):338-353
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The ethylene releasing compound, 2-chloroethylphosphonic acid (ethephon) inhibited nodule development in common bean (Phaseolus vulgaris L.) plants. In contrast, inhibitors of ethylene synthesis or its physiological activity enhanced nodulation. In a co-culture of bean seeds and rhizobia, ethephon inhibited rhizobial growth while inhibitors of ethylene synthesis or action did not influence the growth and proliferation of rhizobia. These data emphasize the role of ethylene as a regulator of nodulation in determinate nodulators and indicate that the ethylene signaling pathway involved in the nodulation process is not limited to the plant host but also involves the bacterial symbiont. 相似文献
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The plant hormone ethylene triggers and enhanced ethylene synthesis in certain ripening fruits and senescing flowers. Unlike most carnation (Dianthus caryophyllus L.) cultivars exhibiting climacteric rise in ethylene production at the onset of senescence, cv. Sandrosa does not show this phenomenon naturally. In order to understand the mechanism of autocatalytic ethylene production, we exposed carnation flowers cv. Sandrosa to ethylene which resulted in an enhanced capacity for ethylene synthesis in the petals. A short time response of one hour was measured for an increase in ACC oxidase activity, about five hours in advance of an increase in ACC synthase activity and ethylene production. The observed enhancement was dependent on the presence of exogeneous ethylene, and could be partially inhibited by prior treatment of the petals with -amanitin or cycloheximide. The results of the present study suggest that in response to ethylene, activation of an existing enzyme is taking place first. This is followed by an increase in expression of ACC oxidase and ACC synthase mRNAs.Abbreviations ACC
1-aminocyclopropane-1-carboxylic acid
- DTT
dithiothreitol
- PMSF
phenyl-methylsulfonyl fluoride
- SAM
S-adenosyl-L-methionine 相似文献
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Yujie Sun Brian Christensen Fulai Liu Huiqun Wang Renate Müller 《Plant Growth Regulation》2009,59(1):83-91
Phalaenopsis frequently exhibits bud drop during production and in response to adverse postharvest conditions. The effect of exogenous
ethylene on bud drop of mini Phalaenopsis was studied and ethylene sensitivity of four cultivars was compared. Water content, membrane permeability and ABA (abscisic
acid) content in floral buds and flowers were determined after ethylene treatment. Exogenous ethylene induced flower bud drop
in all tested Phalaenopsis cultivars and the different cultivars showed distinct differences in ethylene sensitivity. The cultivar Sogo ‘Vivien’ exhibited
the highest bud drop, water loss and change in membrane permeability in floral petals, while Sogo ‘Berry’ showed the lowest
sensitivity. The ethylene inhibitor 1-MCP (1-methylcyclopropene) reduced ethylene-induced floral bud drop in the cultivar
Sogo ‘Yenlin’. ABA content in floral buds was increased in response to ethylene and 1-MCP pretreatment inhibited the ethylene-induced
increase in ABA levels efficiently. This finding suggests that the observed increase in ABA content during bud drop was mediated
by ethylene. The interaction between ABA and ethylene is discussed. 相似文献
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The purpose of this paper is to investigate the differential responses of flower opening to ethylene in two cut rose cultivars,
‘Samantha’, whose opening process is promoted, and ‘Kardinal’, whose opening process is inhibited by ethylene. Ethylene production
and 1-aminocyclopropane-1-carboxylate (ACC) synthase and oxidase activities were determined first. After ethylene treatment,
ethylene production, ACC synthase (ACS) and ACC oxidase (ACO) activities in petals increased and peaked at the earlier stage
(stage 3) in ‘Samantha’, and they were much more dramatically enhanced and peaked at the later stage (stage 4) in ‘Kardinal’
than control during vasing. cDNA fragments of three Rh-ACSs and one Rh-ACO genes were cloned and designated as Rh-ACS1, Rh-ACS2, Rh-ACS3 and Rh-ACO1 respectively. Northern blotting analysis revealed that, among three genes of ACS, ethylene-induced expression patterns of
Rh-ACS3 gene corresponded to ACS activity and ethylene production in both cultivars. A more dramatic accumulation of Rh-ACS3 mRNA was induced by ethylene in ‘Kardinal’ than that of ‘Samantha’. As an ethylene action inhibitor, STS at concentration
of 0.2 mmol/L generally inhibited the expression of Rh-ACSs and Rh-ACO in both cultivars, although it induced the expression of Rh-ACS3 transiently in ‘Kardinal’. Our results suggests that ‘Kardinal’ is more sensitive to ethylene than ‘Samantha’; and the changes
of Rh-ACS3 expression caused by ethylene might be related to the acceleration of flower opening in ‘Samantha’ and the inhibition in
‘Kardinal’. Additional results indicated that three Rh-ACSs genes were differentially associated with flower opening and senescence as well as wounding. 相似文献
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Effect of 1-methylcyclopropene and methylenecyclopropane on ethylene binding and ethylene action on cut carnations 总被引:11,自引:0,他引:11
1-Methylcyclopropene (1-MCP), formerly designated as Sis-X, has been shown to be an effective inhibitor of ethylene responses in carnation flowers in either the light or the dark. The binding appears to be to the receptor and to be permanent. A 6 h treatment at 2.5 nl l–1 is sufficient to protect against ethylene, and 0.5 nl l–1 is sufficient if exposure is for 24 h. As carnation flowers age, a little higher concentration appears to be needed. Most of the natural increase in ethylene production during senescence is prevented by treatment with 1-MCP. A closely related compound, methylenecyclopropane shows ethylene activity. A tritium labelled 1-MCP (60 mCi mmol–1) has been prepared. A higher specific activity is needed for more critical studies. 相似文献
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Kortessa Dimasi-Theriou Athanasios S. Economou Evangelos M. Sfakiotakis 《Plant Cell, Tissue and Organ Culture》1993,32(2):219-225
The influence of ethylene on shoot and root formation from petunia leaf explants was studied in cultures in test tubes placed in 51 glass jars. Reduction of the endogenously produced ethylene by inclusion of ethysorb (KMnO4), an ethylene absorbent, caused a decrease of the number of shoots. On the other hand, supplementing the cultures with ethylene (0.01–10 ppm) caused a marked increase of the number of shoots without, however, any effect on the length and fresh weight. Ethylene treatments (1 ppm) were found to be most effective when they were applied in the second week of culturing of petunia explants. Addition of Co++ to the medium resulted in a reduction of the endogenously produced ethylene and concomitantly reduced shoot formation. Similarly, inclusion of Ag+, an inhibitor of ethylene action, resulted in poor shoot formation. Ethylene also appeared to play a role on rooting of petunia microshoots in vitro in an auxin-free medium. Ethylene at a concentration of 10 ppm induced adventitious root formation considerably, whereas at low levels (0.01–1 ppm) it had no influence on rooting. 相似文献
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The longevity of cut Phalaenopsis (Phalaenopsis hybrid, cv. Herbet Hager) flowers is normally 2 to 3 weeks. After pollination however, there was a rapid acceleration of the wilting process, beginning after only 24 h. Enhancement of senescence in several Phalaenopsis cultivars as well as in Doritaenopsis, Dendrobium and Cymbidium, was induced by successful pollination and only slightly or not at all by emasculation. Wilting of the flowers was accompanied by a loss of water from cells of the upper layer of the petals, leading to their upward folding. Following pollination there was an increase in ethylene production and sensitivity to ethylene. The increase in ethylene production began about 10 h after pollination and reached its peak after 30 h. An obvious increase in sensitivity to ethylene could already be detected 4 h after pollination and reached its peak 10 h after pollination. The increase en ethylene sensitivity following pollination was not dependent on endogenous ethylene production as it occurred also in flowers treated with (aminooxy)acetic acid, an inhibitor of ethylene biosynthesis.Abbreviations AOA =
(aminooxy)acetic acid
- RH =
relative humidity
- SEM =
scanning electron microscope 相似文献
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In all cultivars the force required to harvest fruit declined during ripening. The fruit retention strength (FRS) of ripe fruit varied between cultivars with ‘Ashton Cross’ and ‘Chehalem’ being easier to remove than ‘Bedford Giant’ or ‘Oregon Thornless’. The cultivars ‘Ashton Cross’ and ‘Oregon Thornless’ showed no increase in ethylene production during fruit ripening whereas ‘Bedford Giant’ and ‘Chehalem’ had increased rates of ethylene production (EPR) in the ripe fruit. Exogenous ethylene accelerated abscission, ethylene production and pigment changes in ‘Ashton Cross’ fruit at all stages of development. 1-amino-cyclopropane-1-carboxylic acid (ACC) supplied to fruit at all stages of development was converted to ethylene at levels in excess of those found naturally. The differences between cultivars are discussed with reference to the role of ethylene in both machine harvesting and post-harvest storage of blackberry fruit. 相似文献
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Transport and metabolism of radiolabeled indole-3-butyric acid (IBA) were studied in midrib sections of Cleopatra mandarin (Citrus reticulata Blanco) and compared to that of indole-3-acetic acid (IAA). Exogenous IBA was metabolized by the midribs to a polar compound, probably an ester conjugate. Ethylene pretreatment of the midribs reduced their capacity to metabolize IBA by ca. 70% as compared to air pretreatment. IBA transport capacity in the leaf midribs was ca. two times greater in the basipetal direction than the acropetal. The basipetal transport capacity of 3H-IBA was lower than that of 14C-IAA (ca. 24% and 39% of the uptake, respectively). While ethylene treatment reduced basipetal transport of IAA by ca. 70% it did not affect the transport of IBA. Most of the transported label was found as free IBA, but the reduction of IBA conjugation by ethylene treatment did not affect the transport capacity. 相似文献
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L.?B.?Reis V.?B.?Paiva Neto E.?A.?Toledo Picoli M.?G.?C.?Costa M.?M.?Rêgo C.?R.?Carvalho F.?L.?Finger W.?C.?Otoni
Summary The effects of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) and two inhibitors, silver thiosulfate (STS)
and aminoethoxyvinylglycine (AVG), were tested in yellow passionfruit (Passiflora edulis f. flaricarpa Degener) acillary cultured in vitro. The organogenic responses were assessed by the number of buds per explant, mean leaf area per explant, and shoot length.
ACC-supplemented medium significantly inhibited all evaluated responses at both concentrations tested. When ethylene action
and biosynthesis were inhibited, a significant increase in the number of developed buds and average leaf area was observed.
Accumulated ethylene and its accumulation rate were significantly greater at 10 μM ACC, with a maximum production rate deteeted: at the 14th day and a decline at the 21st day. The results suggest beneficial
effects of ethylene inhibitors on in vitro development of axillary buds and their reliability for use as an alternative approach to evaluate sensitivity of Passiflora species to ethylene. Even though shoot elongation did not differ from that of the control, the inhibition of the ethylene
action and its biosynthesis by AVG and STS, respectively, significantly enhanced the number of buds per explant and leaf area. 相似文献
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Ethylene production, ACC content, and ACC oxidase activity were determined in strawberry fruit harvested at different stages of development and in fruit harvested green and developed in vitro in solutions containing sucrose. In fruit harvested at progressive stages of development from green through full ripe, ethylene production and ACC oxidase activity decreased whereas ACC content increased between the white and pink stages. Fruit detached at the green stage and developed to full ripe by immersion of the cut pedicel in sucrose solutions exhibited an increase in ACC content, decreased ethylene production, and no change in ACC oxidase activity. Detached green fruit provided with sucrose containing 0.5 mM silver (STS) had elevated ethylene production and more ACC oxidase activity than did fruit incubated without the silver salt. Green fruit provided with sucrose containing 1 mM ACC showed markedly increased ACC content, ACC oxidase activity, and ethylene production. These increases were noted following 4 days incubation in ACC, and were more pronounced after 11 days, at which time fruit of all treatments had attained a full-ripe stage of development. Calyx tissue exhibited more ACC oxidase activity, less ACC content, and similar ethylene production compared with receptacle tissue. ACC synthase could not be detected in fruit harvested at different developmental stages or in fruit detached and developed in vitro.abbreviations ACC
1-aminocyclopropane-1-carboxylic acid
- HQS
8-hydroxyquinoline hemisulfate
- SAM
S-adenosyl methionine
- STS
silver thiosulfate 相似文献
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对两个梨品种不同成熟期果实贮藏过程中,整个果实以及果皮、果肉、果心的乙烯释放变化及果肉、种子的钙调素(CaM)含量进行测定。结果表明:(1)黄花品种完整果实及不同部位乙烯释放量都高于耐贮藏的湘南品种,且启动乙烯生成和形成乙烯峰值的时间也早于湘南品种;(2)果实不同部位形成峰值的顺序均依次为果心、果肉、果皮;(3)果实呼吸跃变过程中,CaM含量伴随乙烯释放量的上升而升高,乙烯峰值过后,CaM含量下降,果实衰老。 相似文献
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Improved rates of ethylene oxidation by cell-free preparations from cotyledons of Vicia faba L. have been obtained using cryogenic storage techniques and by developing a method for the hydrolysis of ethylene oxide. Gel permeation chromatography showed that a low-molecular-size fraction was required for activity; accordingly, the kinetics of ethylene oxidation in the presence of this fraction were studied. Reduced pyridine nucleotides could substitute for the low-molecular-size fraction. Activity under a nitrogen atmosphere was 60% lower than in air. The need for reduced nicotinamide adenine dinucleotide phosphate (NADPH) and oxygen indicated that the enzyme might be a mixed-function oxidase. Using sufficient NADPH to approach saturation, the apparent Michaelis constant (K
m) for ethylene was 1.94±0.38 · 10-8 M (aqueous phase), and when ethylene was saturating, the K
m for NADPH was 3.7 · 10-5 M. Carbon monoxide was found to inhibit by competing with ethylene, and the inhibitor constant was 5.97 · 10-7 M in solution. In the presence of excess ethylene and NADPH, activity was highest in phosphate-buffered medium pH 7.9. The bulk of the activity was found in a microsomal fraction.Abbreviations Epps
N-2-hydroxyethylpiperazine-N-3-propane sulphinic acid
- Tris
2-amino-2-(hydroxymethyl)-1,3-porpanediol 相似文献
20.
Jose L. Garcia-Martinez Noah Ben-Shalom Lawrence Rappaport 《Plant Growth Regulation》1984,2(3):209-216
The effect of gibberellin A1 (GA1) on production of ethylene by cowpea (Vigna sinensis cv Blackeye pea no. 5) epicotyl explants and its relationship to epicotyl elongation was investigated. The explants were placed upright in water and incubated in sealed culture tubes or in large jars. GA, and IAA in ethanol solution were injected into the subapical tissues of the decapitated epicotyls. Cowpea epicotyl explants elongated after GA but not after IAA treatment, and they were very sensitive to exogenous ethylene. As little as 0.14 1/1 ethylene reduced significantly GA1-induced epicotyl elongation.Treatment with GA1 induced the production of ethylene which began 10 h after GA application, showed a peak at about 22 h and then declined. The yield of ethylene was proportional to the amount of GA, injected. The inhibition of epicotyl elongation in closed tubes was avoided by absorbing ethylene released with Hg(Cl04)2 , or by adding AVG to the incubation solution to inhibit ethylene production. Treatment with IAA elicited a rapid production of ethylene which ceased about 10 h after application. The effects of IAA and GA1 on ethylene production were additive.Abbreviations AVG
aminoethoxyvinylglycine 2-amino-4-(2-aminoethoxy)-trans-3butenoic acid
- ACC
1-aminocyclopropane-1-carboxylic acid
- GA
gibberellin
- IAA
indole-3-acetic acid 相似文献