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1.
为了研究葡萄不同着色期果皮中蛋白质表达特征,以着色初期、中期和后期等3个着色时期的葡萄果皮为研究对象,运用2-DE、MALDI-TOF/TOF-MS质谱技术以及生物信息学方法对差异蛋白进行分析,结果表明:(1)双向凝胶电泳显示,果皮着色3个时期有1050个高度重复的蛋白点,差异显著的蛋白点有162个,其中108个蛋白得到鉴定,有87个蛋白映射到葡萄蛋白质组数据库中;3个时期均表达的差异蛋白有20个,且随着果皮颜色加深,差异蛋白数量呈上升趋势。(2)GO分析显示,ATP合成酶β亚基(ATPβ)极显著富集于磷酸核糖代谢过程,对着色初期果皮生理代谢的能量需求具有重要意义。(3)KEGG分析表明,碳代谢、生物固碳、磷酸戊糖、氨基酸生物合成等代谢通路在果皮着色的不同时期显著富集。(4)qRT-PCR分析显示,S–腺苷甲硫氨酸合成酶基因(VvMETK4)在果皮着色后期表达量最高。  相似文献   

2.
VFL and VvTFL1 genes expression patterns and the effects of sucrose on the expression of VFL and VvTFL1 genes in different organs of the “Xiangfei” grapevine (Vitis vinifera L.) were investigated. VvTFL1 gene expression was detected in the meristem of the apical bud and lateral bud, but was not detected during inflorescence differentiation and flower organ development. After sucrose treatment, VvTFL1 gene expression increased in the apical bud, but decreased in the lateral bud. These results suggested that the VvTFL1 gene might be mainly involved in the apical growth process of shoots, and exogenous sucrose had an effect on the VvTFL1 gene by increasing shoot apical meristem initiation of apical buds. The VFL gene was expressed primarily during inflorescence differentiation and early flower organ development, but it gradually reduced in later flower development. After sucrose treatment, VFL gene expression increased in the inflorescence and small or middle flower, but a little change was seen in the large flower. These results suggested that the VFL gene plays important roles in the initiation of inflorescence meristems and the morphological formation of flower organs. Exogenous sucrose had an effect on VFL gene expression at the early stage of flower development.  相似文献   

3.
AIM:To identify the proteins related to aging in the mitochondria of the heart. METHODS:Mitochondria were isolated from the hearts of adult (10-week) and old (12-month) rats (n=3). Total proteins extracted from the mitochondria were applied to the two-dimensional gel electrophoresis. The differentially expressed protein spots evaluated by a software were subjected to in-gel digestion, and analyzed by the matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). ATP content in the heart was also measured. RESULTS:Eighty-four differentially expressed protein spots were observed, and fifteen of them were analyzed by MALDI-TOF- MS. Four proteins including creatine kinase, peroxiredoxin 2, Tu translation elongation factor and isocitrate dehydrogenase were up-regulated and 11 proteins including succinate dehydrogenase, malate dehydrogenase, aconitate hydratase, NADH dehydrogenase, ATP synthase H+ transporting, ATP synthase beta chain, heat shock protein 60, glucose-regulated protein 78, prohibitin, Aldehyde dehydrogenase 2 and voltage-dependent anion channel exhibited down-regulation in the aging group compared to the adult one. ATP content in the heart of old rats was significantly reduced as compared to that in adult rats (n=5, P<0.01). CONCLUSION:Significant changes in the mitochondrial protein expression in the aging heart were identified by the 2-DE electrophoretogram with high resolution and reproducibility. The functions of these identified proteins need to be further investigated.  相似文献   

4.
Haploid production using in vitro ovule cultures has long been recognized as an important tool to produce haploid and homozygous double-haploid plants for genetic studies and plant breeding programs. In the present study, four experiments were carried out to study the influence of genotype, position of female flowers on plant stem, temperature and sucrose concentration on the in vitro gynogenesis induction of squash. (1) Ovules of 12 genotypes were excised from female flowers, 1 day before anthesis, and cultured onto MS medium containing 3% sucrose and 1 mg l−1 from each of kinetin and 2,4-D (2,4- dichlorophenoxy acetic acid). Differences in response among genotypes were demonstrated. Raad F1 showed the highest percentage of responding ovules and number of plantlets per dish with 48.8% and 15 plants, respectively. The results revealed that genotype is a key factor influencing the in vitro gynogenesis in squash. (2) Ovules were excised from first, second and third female flower of two hybrids (Giad and Raad) and cultured onto the mentioned above medium. The highest percentage of responding ovules and number of plantlets per dish were obtained from ovules excised from the second female flower on the plant stem. (3) Effect of temperature (4 and 32 °C) for 0, 4, 7 and 12 days on the ovule culture of Queen F1 was studied. Ovules incubated at 4 or 32 °C for 4 days produced a better embryogenic response. (4) Three sucrose concentrations (30, 60 and 90 g l−1) were tested with the ovule cultures of the local cultivar (Eskandrani). Differences among sucrose concentrations were statistically significant and ovules cultured on the MS medium containing 30 g l−1 produced the best result. MS medium containing 90 g l−1 did not produce gynogenic ovules.  相似文献   

5.
A suitable protocol for in vitro tuber production using non-dormant tubers of Gloriosa superba L. on Murashige and Skoog (MS) medium without addition of plant growth regulators is reported in the present study. Among the different basal media tested MS medium was found to be suitable for induction and development of secondary tubers; one in vitro tuber per explant was obtained after 6 weeks and 3 tubers per explant after 12 weeks of culture. These tubers produced healthy green shoots that rooted on basal medium. Best rooting was noted in half strength (half organics and inorganics) MS medium with one-fifth nitrates.  相似文献   

6.
In vitro symbiotic seed germination is an important tool not only for the study of orchid-fungus specificity but also for the production of mycobiont-infected healthy seedlings that could be valuable for both horticultural and conservation purposes. The current study compared effectiveness of eight putative orchid mycorrhizal fungi obtained from mature orchids in the genera Paphiopedilum, Cymbidium and Dendrobium, in promoting in vitro seed germination and protocorm development of Grammatophyllum speciosum Blume and Dendrobium draconis Rchb. f., native Thai orchids. The developmental stages of seeds and protocorms cultured on Murashige and Skoog (MS) medium, oat meal agar (OMA), or OMA inoculated with one of the eight fungal isolates were evaluated weekly. Two isolates of Epulorhiza repens (Bernard) Moore (=anamorphic species of Tulasnella calospora (Boud.) Juel), Da-KP-0-1 and Pv-PC-1-1, were found to be the most effective fungi in promoting protocorm development of G. speciosum. At week 13, protocorms co-cultured with either one of these two fungal isolates, on the average, were significantly more advanced than those sown on OMA. Protocorms co-cultured with isolate Pv-PC-1-1 were also significantly more advanced than those cultured on MS medium. For D. draconis seed germination, three fungal isolates of different anamorphic species of Tulasnella, C1-DT-TC-1, Pv-PC-1-1, and C3-DT-TC-2, were found to be the most effective fungi in promoting protocorm development. However, none of these fungal isolates outperformed MS medium. Additionally, the compatibility between the fungal isolates tested and the two orchid species was discussed.  相似文献   

7.
Our objective was to study the differentially expressed proteins (DEP) in various Malus spectabilis (crabapple) varieties (M. ‘Snowdrift’, M. ‘Hongling’ and M. ‘Hongjin’) in relation to Malus domestica (‘Gala’) and their role in pollination. Our method used a two-dimensional electrophoresis (2-DE) to analyse the differential proteins in the pollen of several crabapples. The 2-DE apples combined with the tandem mass spectrometry (MS-MS) and protein database retrieval helped us to identify the nature and function of DEPs in ‘Gala’ apples and crabapples. We identified 1195 proteins through 2-DE. Among these, six DEPs, namely chloroplast ferritin, Actin, Beta-fructofuranosidase, vacuolarH+-ATPase catalytic subunit, Full = Phosphoglucomutase, and Cytochrome b were identified by MS-MS. This study identified six DEPs among the pollen from the ‘Snowdrift’ crabapple, ‘Hongling’ crabapple, ‘Hongjin’ crabapple, and ‘Gala’ apples. The DEPs included metabolism related proteins, stress/regulatory proteins, and proteins involved in signal transduction.  相似文献   

8.
'白核'龙眼种子败育不同时期差异蛋白分析   总被引：1，自引：0，他引：1  
以'白核'龙眼种子为试验材料,比较两种蛋白质的提取方法,确定了酚抽法提取龙眼种子蛋白.应用蛋白质组学研究技术,分析了龙眼种子败育4个不同时期的蛋白质组变化,共发现21个差异蛋白,其中9个上调表达,12个下调表达.通过MALDI/TOF/TOF-MS/MS质谱分析和蛋白质数据库检索,共鉴定出12个差异蛋白,分别为胞质苹果酸脱氢酶、丙酮酸脱氢酶、磷酸甘油酸变位酶、RuBisCO大亚基结合蛋白α、β亚基、17 kD HSP、抗坏血酸过氧化物酶、胞质抗坏血酸过氧化物酶、半胱氨酸蛋白酶、Dessication-related protein以及两个未知蛋白.这些鉴定出的差异蛋白质与能量和物质代谢、分子伴侣功能、自由基清除和抗氧化作用、细胞凋亡等生理过程密切相关,可能参与了龙眼种子败育过程.  相似文献   

9.
The regenerability of three ornamental species—Lysimachia christinae, Lysimachia rubinervis and Lysimachia nummularia ‘Aurea’, were investigated using in vitro leaves and shoot tips. 6-Benzylaminopurine (BAP) and α-naphthalene acetic acid (NAA) added to Murashige and Skoog (MS) medium were tested for their effect on organogenesis. On the medium, shoot regeneration occurred directly without callus formation. In these species, L. christinae developed the highest regeneration rate and numbers of shoots/explant from shoot tips (100%, 12.25) and leaf bases (100%, 13.01) on the MS medium containing 3.0 mg l−1 BAP and 0.1 mg l−1 NAA. For L. rubinervis, the highest shoot induction rate and number of shoots/explant were obtained from shoot tip (100%, 16.87–17.20) on the MS medium with 0.1 mg l−1 NAA and 3.0–5.0 mg l−1 BAP. L. nummularia ‘Aurea’, however, showed the highest regeneration rate and number of shoots/explant (100%, 12.73) from leaf bases on MS medium supplemented with 1.0 mg l−1 BAP and 0.1 mg l−1 NAA. All in vitro shoots rooted well on half macronutrient MS medium containing 0.1 mg l−1 NAA. After acclimatization, transplanted plantlets grew normally and flowered in the field.  相似文献   

10.
The present study reports that protoplasts isolated from stoloniferous shoots (SS) of potato represent an efficient system for somatic cell genetic manipulations. SS were established from single-node cuttings on MS medium supplemented with either 0.1 or 0.2 M sucrose (Suc), and protoplasts were isolated and cultured within the alginate strip, following an improved method. SS induced by 0.1 M Suc yielded 8–22 × 105 protoplasts g−1 fresh mass, with a high morphogenic competence. However, 0.2 M Suc-induced SS yielded protoplasts that contained large amounts of starch grains, resulting in their high degree of fragility, delayed cell division and poor morphogenic competence. For symmetric somatic hybridization (electrofusion) between Solanum tuberosum Gp. Tuberosum androgenic (di)haploid (2n = 2x = 24) ‘C-13’ and diploid (2n = 2x = 24) wild species S. pinnatisectum, protoplasts isolated from 0.1 M Suc-induced SS were also found to be most responsive. Out of several putative somatic hybrids, there were two tetraploids and five diploids, with 48 and 24 chromosomes, respectively at all the three shoot layers (L1–L3). This precluded the occurrence of mixoploidy vis-à-vis chimaerism in regenerants, as common in somatic fusion involving mesophyll protoplasts of S. pinnatisectum. Nuclear microsatellite analyses based on the two single-locus nSSR loci (STM0037 and STM2030) confirmed that one of the tetraploids was a true nuclear hybrid (heterokaryon), while the other a homokaryon of the Tuberosum parent ‘C-13’. The use of 0.2 M Suc-induced SS protoplasts for fundamental studies on tissue- and/or cell type-specific transient gene expression underlying tuberization has been discussed.  相似文献   

11.
Shoots regenerated adventitiously on epicotyl segments from in vitro seedlings of Emblica officinalis var. ‘Kanchan’. Epicotyls derived from 2-week-old aseptic seedlings were most responsive and produced a maximum number of 303 shoots per explant in Murashige and Skoog (1962) medium (MS) augmented with 8.8 μM N6-benzyladenine (BA) + 1.425 μM indole-3-acetic acid (IAA). Shoots readily elongated in MS lacking growth regulators and rooted in half-salt-strength MS (1/2 MS) supplemented with indole-3-butyric acid (IBA) or α-naphthalene acetic acid (NAA). The highest rooting response was recorded in 1/2 MS containing 14.7 μM IBA. Plantlets were acclimatized inside the green house and 80% of the plantlets survived on transfer to garden soil.  相似文献   

12.
Inter-section hybrids were obtained in the reciprocal crosses between Primula filchnerae (2n = 2x = 24) of Sect. Pinnatae and P. sinensis ‘Fanfare’ (2n = 2x = 24) of Sect. Auganthus by rescuing ovules on half-strength (1/2) Murashige and Skoog's (MS) medium supplemented with 50 g l−1 sucrose, 2.5 g l−1 gellan gum, 0.1 mg l−1 α-naphthaleneacetic acid (NAA), 0.1 mg l−1 6-benzyladenine (BA) and 50 mg l−1 gibberellic acid (GA3). In ovule culture, germination occurred with radicle elongation but no plumule was observed. The radicle kept on the initial medium showed root proliferation with callus formation. When the calluses were transferred to (1/2)MS media containing 30 g l−1 sucrose and 3 g l−1 gellan gum, without plant growth regulators (PGRs) or with 1 mg l−1 zeatin and 0.1 mg l−1 NAA, plantlets were regenerated. The plants thus obtained were confirmed to be hybrids through flow cytometry (FCM) and random amplified polymorphic DNA (RAPD) analyses. The hybrid obtained when P. filchnerae was used as the maternal parent was diploid, whereas hexaploid hybrid was obtained when using P. sinensis as the maternal parent. The hexaploid hybrid might be produced through chromosome doubling of a triploid originated from the fertilization of P. sinensis with unreduced pollen of P. filchnerae.  相似文献   

13.
试管芋诱导的研究   总被引：12，自引：0，他引：12  
 以芋组培苗为试材, 研究了蔗糖浓度、激素浓度、光照时间、培养温度、不同大小试管苗,不同芋品种类型等因素对试管芋诱导的影响及不同基质对试管芋育苗成活率的影响。结果表明: 诱导试管芋较理想的培养基为MS + 蔗糖8 % + BA 1. 0 mg·L - 1 + NAA 0. 5 mg·L-1 , 光照12 h/ d , 培养温度30 ℃; 试管苗越大越有利于试管芋的形成; 试管芋可保持其原有的特性; 基质影响试管芋育苗的成活率, 4 种基质中蛭石最好。  相似文献   

14.
We present here a method for the regeneration of the prickly-pear (Opuntia ficus-indica (L.) Mill.) through somatic embryogenesis (SE). Direct SE was induced by cultivating shoot apices devoid of leaf primordia on semisolid Murashige and Skoog (MS) basal medium supplemented with 4-amino 3,5,6-trichloropicolinic acid (picloram) at 4 mg l−1. Somatic embryogenesis was influenced by the type, age, physiological and developmental stage of the explants, and also by light conditions, wounding, and the sucrose concentration in the induction medium. A histological analysis confirmed SE by revealing the presence of a closed vascular system in the developing embryos and the absence of a vascular connection between the somatic embryos and the explant.  相似文献   

15.
A proteomic approach was taken to compare the proteomes of normal flowering buds and flowering reversion buds in longan (Dimocarpus longan Lour. cv. Longyou). Two-dimensional gel electrophoresis (2-DE), coupled with mass spectroscopy and protein database searching, recognized 18 proteins that were differentially expressed in flowering reversion buds. Eleven of these were down-regulated, whereas seven were up-regulated. A subset of 13 proteins was identified by MALDI-TOF-TOF/MS and classified into regulatory proteins (kinase, 20S proteasome alpha 6 subunit, putative alpha 7 proteasome subunit, auxin-induced protein, and abscisic stress ripening-like protein), antioxidant-related proteins (Chain A, GDP-mannose-3′,5′-epimerase, putative lactoylglutathione lyase, and Chain A, ascorbate peroxidase), pollen fertility-related proteins (putative leucoanthocyanidin reductase 2, and putative isoflavone reductase), photosynthesis-related proteins (large subunit, ribulose-bisphosphate carboxylase–oxygenase), and molecular chaperones (disulfide isomerase). Among them, regulatory and antioxidant-related proteins accounted for almost two-thirds of these proteins, suggesting that they may play a more important role in bud differentiation. Identification of these proteins provides insights that may lead to a better understanding of the molecular basis for flowering reversion in longan.  相似文献   

16.
A protocol is developed for Agrobacterium-mediated genetic transformation of Amaranthus tricolor via explant co-cultivation with Agrobacterium rhizogenes. Bacteria-plant specific factors which influenced transformation were optimized. Of the two Agrobacterium strains employed, LBA9402 was more infectious compared to A4. Bacterial suspensions grown overnight with 100 μM acetosyringone and experiencing O.D.660 = 0.6 followed by dilution to a density of 109 cells ml−1 were the most effective. Explants from garden-grown plants were more responsive than those from in vitro cultures; stem internodes being better than leaves. Immersion of the pre-pricked explants in bacterial suspension resulted in a markedly higher transformation frequency compared to the direct injection method. The infection of internode explants with the LBA9402 strain followed by co-cultivation on growth regulator-free MS medium (MS0) for 5 days resulted in emergence of hairy roots up to a maximum frequency of 97.22%. Roots were individually cultured in MS0, but fortified with bactericidal antibiotic (500 μg ml−1 cefotaxime). Rhizoclones showing prolific growth were renewed through successive subcultures in MS0. Opine gene expression was revealed by positive agropine and mannopine synthesis in all selected transformed rhizoclones. Shoot regeneration from root clones, capable of auxin-independent growth and opine proficiency, was stimulated in MS augmented with 2.0 mg l−1 zeatin. pRi TL–DNA rolB and pRi TR–DNA man2 ORF were detected in leaf tissues of regenerated plants from selected hairy root clones through PCR amplification. The implication of such findings is discussed on the possibility of conferring protection to crop amaranths against biotic stress challenges, particularly due to insects, viruses or fungal pathogens.  相似文献   

17.
利用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)标记结合二维液相色谱串联质谱(two-dimensional liquid chromatography tandem mass spectrometry,2D LC-MS/MS)技术,研究广叶绣球菌(Sparassis latifolia)原基期、幼菇期和成熟子实体阶段的差异表达蛋白质组.采用Q-Exactive质谱鉴定并经ProteinPilot软件搜库,对所获得的差异蛋白进行GO (gene ontology)、KEGG(kyoto encyclopedia of genes and genomes)和转录因子注释分析.结果共鉴定到可信蛋白2305个,其中2219个蛋白具有相对定量信息.与原基期相比,幼菇期显著上调蛋白104个,下调蛋白142个,子实体阶段显著上调蛋白155个,下调蛋白460个.GO分子功能提示这些差异性蛋白主要参与催化活性、蛋白结合和水解酶活性.KEGG代谢通路分析结果显示,差异蛋白主要涉及碳代谢、氨基酸合成、核糖体、糖酵解/糖衍生等代谢过程.差异蛋白中与信号传导和转录因子相关的蛋白数量分别为27个和7个.  相似文献   

18.
Synseeds of ginger (Zingiber officinale) were produced using aseptically proliferated 2-week old encapsulating explants (microshoots) upon complexation of 4% sodium alginate prepared in Murashige and Skoog (1962) medium (MS) and 100 mM calcium chloride. Conversion of synseeds into plantlets (conversion) was recorded as 66% and 53% on MS (3% sucrose) and on MS (3% sucrose) + 2.5 mg/l BA media, respectively. However, shoots/synseed were significantly higher on MS (3% sucrose) + 2.5 mg/l BA medium. For short-term storage of germplasm, sucrose-dehydrated synseeds were found better than air-dehydrated or fresh synseeds. Synseeds dehydrated in 0.25 M sucrose liquid medium for 16 h and stored in cryovials (with out medium) at 25 °C for 8 weeks and 12 weeks exhibited 53% and 13% conversion, respectively, on MS (3% sucrose) + 2.5 mg/l BA medium. Plantlets obtained from stored synseeds were hardened, established successfully ex vitro and were morphologically similar to each other as well as their mother plants. This synseed protocol could be useful for short-term storage and exchange of germplasm of ginger between national as well as international laboratories.  相似文献   

19.
Kosteletzkya virginica, a perennial dicot halophytic species of the Malvaceae, is native to American salt marsh. It was introduced into China as a potential species to improve coastal wetlands and to develop ecologically sound saline agriculture. K. virginica adapts excellently to the tidal-flat habitats in China's east coast, with multiple eco-benefits; in particular, its seed oil could be used to produce biodiesel. The purpose of this study was thus to develop a standardized protocol to induce a high frequency of callus and subsequent plantlet regeneration system for a K. virginica breeding program with the final objective of applying transgenic techniques to improve seed oil yield. The embryonic axes of K. virginica were used as explants for callus induction, shoot induction from the callus and then adventitious root induction from the shoots on nine culture media with different hormone combinations. The best results were achieved on the following media: (1) 93.94% callus induction on MS medium supplemented with 1.0 mg L−1 indole-3-acetic acid (IAA), 0.3 mg L−1 kinetin, 30 g L−1 sucrose and 8 g L−1 agar; (2) 65.83% shoot induction on 1/2MS medium supplemented with 0.1 mg L−1 IAA, 0.5 mg L−1 zeatin, 30 g L−1 sucrose and 8 g L−1 agar; (3) 96.67% rooting on MS medium containing 30 g L−1 sucrose and 8 g L−1 agar. The survival rate of plantlets by organogenic regeneration was 85% after being transplanted into potting soil in flowerpots and placed in the greenhouse. This experiment indicates that we established successful callus induction and plant regeneration protocols for K. virginica.  相似文献   

20.
Anthracnose, caused by Colletotrichum gloeosporioides, is a major post-harvest disease in papaya fruit. The major objectives of the present work were to isolate, select and test the in vitro and in vivo ability of epiphytic microorganisms, isolated from papaya fruit and leaf surfaces, in controlling anthracnose onset after harvest. A total of 75 bacteria, 67 yeasts and 22 mycelial fungi were isolated. Thirty yeast isolates were able to inhibit the mycelial growth of C. gloeosporioide in vitro and seven of those were used in in vivo assays, resulting in the identification of two very effective isolates. Isolate CEN63, identified molecularly as Cryptococcus magnus, was the most effective in controlling the disease and therefore was studied in more detail. The results of the assays with C. magnus provided evidence that when fruit were treated with the antagonists at concentrations of 107 to 108 cells/ml, as early as 24 h, preferentially 48 h, before inoculation with the pathogen, the development of disease was significantly reduced. C. magnus is a potential antagonist for the development of a commercial product. Additional studies on the modes of action of this yeast isolate, as on its ability to interact with fungicides are being conducted to generate solid basis for the development of an environmentally friendly control agent.  相似文献   

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