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1.
Endogenous gibberellin (GA)-like substances were examined in suspension cultures of somatic embryos of a hybrid grape (Vitis vinifera × Vitis rupestris) during embryogenesis, and in mature embryos chilled at 4°C, and subsequently incubated at 26°C with and without abscisic acid (ABA). The extract was separated into a nonpolar fraction (would contain GA-precursors); a fraction that would contain free GAs; and a highly H2O-soluble fraction (would contain GA glucosyl conjugates and very polar free GAs). Quantitation after SiO2 partition chromatography was accomplished by microdrop and immersion dwarf rice bioassays. As embryogenesis developed, the free and highly H2O-soluble GA-like substances, expressed on a dry weight basis, decreased (however, they increased on a per embryo basis). Chilling at 4°C for 1 week greatly increased activity of free GA-like substances (per g dry weight and per embryo), it then declined over the next three weeks of chilling. Activity (per g dry weight and per embryo) in the H2O-soluble fraction declined throughout chilling. Activity in the GA-precursor fraction, however, increased steadily with chilling (per g dry weight and per embryo). Incubation at 26°C after chilling enhanced activity in the free GA and H2O-soluble fractions (per g dry weight and per embryo), but activity in the GA-precursor fraction dropped dramatically. Incubation at 26°C with (±) ABA after chilling prevented germination and maintained high activity for GA precursors and less polar free GAs and low activity in the polar free GA and H2O-soluble fractions.

Kaurene and kaurenoic acid were characterized in the GA-precursor fraction of chilled embryos by gas-liquid chromatography-mass spectrometry (GLC-MS). The existence of GA4 and GA9 in ABA-treated, chilled embryos was also confirmed by GLC-MS.

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2.
葡萄体细胞胚的保存   总被引:3,自引:0,他引:3  
研究干化对酿酒葡萄品种'神索'体细胞胚保存效应的结果表明,葡萄体细胞胚失水量在40%~50%之间的萌发率较高,达58%左右,比未经干化的萌发率高9.4%;在较高相对湿度的情况下,体细胞胚失水速度变慢,干化处理时间延长,可以提高体细胞胚的存活率和萌发率;相对湿度70%的情况下干化15 d的效果最好,其萌发率从未经干化处理的33.5%提高到56.2%.  相似文献   

3.
Field pennycress (Thlaspi arvense L.) is a winter annual crucifer with a cold requirement for stem elongation and flowering. In the present study, the metabolism of exogenous [2H]-ent-kaurenoic acid (KA) and [14C]-gibberellin A12-aldehyde (GA12-aldehyde) was compared in thermo- and noninduced plants. Thermoinduction greatly altered both quantitative and qualitative aspects of [2H]-KA metabolism in the shoot tips. The rate of disappearance of the parent compound was much greater in thermoinduced shoot tips. Moreover, there was 47 times more endogenous KA in noninduced than in thermoinduced shoot tips as determined by combined gas chromatography-mass spectrometry (GC-MS). The major metabolite of [2H]-KA in thermoinduced shoot tips was a monohydroxylated derivative of KA, while in noninduced shoot tips, the glucose ester of the hydroxy KA metabolite was the main product. Gibberellin A9 (GA9) was the only GA in which the incorporation of deuterium was detected by GC-MS, and this was observed only in thermoinduced shoot tips. The amount of incorporation was small as indicated by the large dilution by endogenous GA9. In contrast, thermo- and noninduced leaves metabolized exogenous [2H]-KA into GA20 equally well, although the amount of conversion was also limited. These results are consistent with the suggestion (JD Metzger [1990] Plant Physiol 94: 000-000) that the conversion of KA in to GAs is under thermoinductive control only in the shoot tip, the site of perception for thermoinductive temperatures in field pennycress. There were essentially no differences in the qualitative or quantitative distribution of metabolites formed following the application of [14C]-GA12-aldehyde to the shoot tips of thermo- or noninduced plants. Thus, the apparent thermoinductive regulation of the KA metabolism into GAs is probably limited to the two metabolic steps involved in converting KA to GA12-aldehyde.  相似文献   

4.
Immature seeds of apricot (Prunus armeniaca L.) were fed the native gibberellin A5 (GA5) as 1- and 1,2-[3H]GA5 (5.3 Curies per millimole to 16 milliCuries per millimole) at doses (42 nanograms to 10.6 micrograms per seed) 2 to 530 times the expected endogenous level. After 4 days of incubation, seeds were extracted and free [3H]GA-like metabolites were separated from the highly H2O-soluble [3H]metabolites. For high specific activity feeds the retention times (Rts) of radioactive peaks were compared with Rts of authentic GAs on sequential gradient-eluted → isocratic eluted reversed-phase C18 high performance liquid chromatography (HPLC) -radiocounting (RC). From high substrate feeds (530 and 230 × expected endogenous levels) HPLC-RC peak groupings were subjected to capillary gas chromatography-selected ion monitoring (GC-SIM), usually six characteristic ions. The major free GA metabolites of [3H] GA5 were identified as GA1, GA3, and GA6 by GC-SIM. The major highly water soluble metabolite of [3H]GA5 at all levels of substrate GA5 had chromatographic characteristics similar to authentic GA1-glucosyl ester. Expressed as a percentage of recovered radioactivity, low substrate [3H]GA5 feeds (2 × expected endogenous level) yielded a broad spectrum of metabolites eluting at the Rts where GA1, GA3, GA5 methyl ester, GA6, GA22, GA29 (17, 14, 1.6, 7, 1.1, 0.5%, respectively) and GA glucosyl conjugates of GA1, GA3, GA5, and GA8 (33, 11, 1, 0.1%, respectively) elute. Metabolites were also present at Rts where GA glucosyl conjugates of GA6 and GA29 would be expected to elute (8 and 0.1%, respectively). Only 5% of the radioactivity remained as GA5. Increasing substrate GA5 levels increased the proportion of metabolites with HPLC Rts similar to GA1, GA6, and especially GA1 glucosyl ester, primarily at the expense of metabolites with HPLC Rts similar to GA3, GA3-glucosyl ester, and a postulated conjugate of GA6. There was evidence that high doses of substrate GA5 induced new metabolites which often, but not always, differed from GA1, GA3, and GA6 in HPLC Rt. These same metabolites, when analyzed by GC-SIM yielded m/e ions the same as the M+ and other characteristic m/e ions of the above GAs, albeit at differing GC Rt and relative intensities.  相似文献   

5.
Histological steps of callogenesis and proembryogenesis in anthercultures ofVitis vinifera L. ‘Grenache noir’ aredescribed. Embryogenic calli were obtained on Nitsch and Nitschmedium supplemented with 1mgl-12,4-dichlorophenoxyacetic acid(2,4-D) and 0.25mgl-1benzylaminopurine (BAP). Calli were initiatedfrom anther connective cells only and no division of microsporesoccurred. The embryos were hence of somatic origin. Proembryosdeveloped either directly (i.e. without intervening callus)from the endothecium, or indirectly from the connective-derivedcallus. In both cases, proembryos originated from single cells.They developed from starchy differentiated cells of a predeterminedtype. The polarity of the somatic proembryo was establishedfrom the first divisions and it was marked by precocious developmentof an easily recognizable suspensor. Other analogies with thedevelopment of the zygote are also emphasised. Vitis vinifera L.; grapevine; somatic embryogenesis; proembryogenesis; histology  相似文献   

6.
Uptake and metabolism of 1,2-[3H]gibberellin A1 ([3H]GA1, I) and its 3-hydroxy epimer ([3H]pseudoGA1, II) by barley (Hordeum vulgare L.) half-seeds were measured after 24 hours of incubation, in the presence or absence of abscisic acid in the media. Uptake of both compounds was enhanced by abscisic acid, and abscisic acid enhanced the extent of metabolism of [3H]GA1. However, [3H]pseudoGA1 was not metabolized, even in the presence of abscisic acid. The significance of the stereo-chemistry of the 3-hydroxyl position is discussed.  相似文献   

7.
After 30 minutes of incubation of young leaf sections of d-5 maize (Zea mays L.) in [3H]gibberellin A1 ([3H]GA1), the metabolite [3H]GA8 was present in significant amounts, with a second metabolite, [3H]GA8-glucose ([3H]GA8-glu), appearing soon after. A third [3H]GA1 metabolite, the polar uncharacterized conjugate [3H]GA1-X, took more than 1 hour to appear. The protein synthesis inhibitor cycloheximide inhibited the production of all [3H]GA1 metabolites, indicating a possible protein synthesis requirement for [3H]GA1 metabolism.  相似文献   

8.
Somatic embryos ofVitis vinifera L. ‘Grenache noir’develop abnormally and form viable plantlets at very low frequencies.They grow continuously and, after the torpedo stage, they formgiant structures which do not undergo further organogenesis.Morphological, histological and cytochemical data were usedto study development from the globular to the giant-embryo stage.Histological organization of somatic embryos until the torpedostage was similar to that of zygotic embryos. Somatic embryosformed bipolar axes, which differentiated precociously and simultaneouslya root and a shoot meristem. However, they differed from theirzygotic homologues by forming a cotyledonary crown or multiplecotyledons and by their rapid cellular differentiation. At theend of the torpedo stage and up to the giant-embryo stage, somaticembryos underwent some characteristic events of germination:the radical grew, tannins accumulated, and protodermal cellssuberized. However the shoot apex was rapidly disorganized anddisappeared. This peculiar behaviour is discussed in comparisonwith the phenomenon of precocious germination often observedfor immature zygotic embryos inin vitro culture. Vitis vinifera ; grapevine; somatic embryo development; precocious germination; histology  相似文献   

9.
Winter canola (Brassica napus cv Crystal) is an oilseed crop that requires vernalization (chilling treatment) for the induction of stem elongation and flowering. To investigate the role of gibberellins (GAs) in vernalization-induced events, endogenous GA content and the metabolism of [3H]GAs were examined in 10-week vernalized and nonvernalized plants. Shoot tips were harvested 0, 8, and 18 d postvernalization (DPV), and GAs were purified and quantified using 2H2-internal standards and gas chromatography-selected ion monitoring. Concentrations of GA1, GA3, GA8, GA19, and GA20 were 3.1-, 2.3-, 7.8-, 12.0-, and 24.5-fold higher, respectively, in the vernalized plants at the end of the vernalization treatment (0 DPV) relative to the nonvernalized plants. Thermoregulation apparently occurs prior to GA19 biosynthesis, since vernalization elevated the concentration of all of the monitored GAs. [3H]GA20 or [3H]GA1 was applied to the shoot tips of vernalized and nonvernalized plants, and after 24 h, plants were harvested at 6, 12, and 15 DPV. Following high-performance liquid chromatography analyses, vernalized plants showed increased conversion of [3H]GA20 to a [3H]GA1-like metabolite and reduced conversion of [3H]GA1 or [3H]GA20 to polar 3H-metabolites, putative glucosyl conjugates. These results demonstrate that vernalization influences GA content and GA metabolism, with GAs serving as probable regulatory intermediaries between chilling treatment and subsequent stem growth.  相似文献   

10.
通过同时测定葡萄木质部导管液流平均流量和流速,应用变形的哈根-泊萧叶方程,研究了测定葡萄木质部压力梯度的新方法。结果表明,在直径分别为7mm的葡萄茎段和4mm的葡萄叶柄上,用此新方法测得的木质部压力梯度与实际加在其上的压力梯度吻合的很好,说明此新方法可用于连续、实时测定葡萄木质部压力梯度。用此新方法在离体葡萄枝上测定发现,随光照强度增强,其木质部压力梯度成线性正相关增加。  相似文献   

11.
根域限制对葡萄果实发育和主要营养物质含量的影响   总被引:2,自引:0,他引:2  
根域限制下葡萄果实在第二次快速生长期间糖分积累显著提高,有机酸含量下降更明显.随着果实pH值的升高,果实发育的第二次快速生长期间花青素含量迅速增加,而且根域限制的高于未受根域限制的.在果实的第二次快速生长期间葡萄果实中总蛋白和游离氨基酸含量迅速增加,也是根域限制的高于未受根域限制的.开花后30~40 d,维生素C含量达到最大,随后下降,果实发育后期又升高,而且根域限制的葡萄果实中的维生素C含量一直高于根域未受限制的.  相似文献   

12.
在PEG引起的水分胁迫下,葡萄试管苗生长发育迟滞,植株节间变短,株高变矮,但根数增加,根变长;新生叶片小而皱缩,气孔指数降低,高浓度PEG(6%)处理的植株叶片上皮细胞排列更紧凑.  相似文献   

13.
Native polyacrylamide gel electrophoresis (PAGE) was employed to show the highest number of esterase loci and to detect alpha- and beta-esterase polymorphisms in leaf buds of Vitis vinifera cultivars. A total of 16 esterase isozymes were detected in leaf buds from 235 plants including Italia, Rubi, Benitaka, and Brasil cultivars. Biochemical characterization of the grape esterases using ester substrates revealed alpha-, beta-, and alpha/beta-esterases with inhibitor tests distinguishing both carboxylesterases (EST-2, EST-3, EST-5, EST-6, EST-7, EST-8, EST-9, EST-10, and EST-16 isozymes) and acetylesterases (EST-4, EST-11, EST-12, EST-13, EST-14, EST-15 isozymes). No allele variation for alpha-, beta-, and alpha/beta-esterases was detected; however, EST-3 alpha-carboxylesterase was absent in 61.7% of vines, and EST-4 alpha/beta-acetylesterase was absent in one vine of Rubi cv. Null EST-3 carboxylesterase phenotype (61.7%) cannot be explained in this article, but the high genetic polymorphism in four V. vinifera clones is a positive aspect for genetic selection and development of new clones with different characteristics.  相似文献   

14.
Gibberellin A4 (GA4) can substitute for light in the germination of Grand Rapids lettuce seeds. Seeds imbibed in [3H]GA4 do not convert this to other GAs prior to, or immediately following, visible germination: thus GA4 alone can promote radicle expansion. Abscisic acid inhibited [3H]GA4-induced germination, but did not significantly affect [3H]GA4 uptake or metabolism during germination. 6N-benzyladenine overcame the inhibitory effect of abscisic acid and increased [3H]GA4 uptake, although radicle emergence was delayed somewhat.  相似文献   

15.
Leaves from annual young grape plants (Vitis vinifera L. cv. Jingxiu) were used as experimental materials. The ultrastructural characteristics of mesophyll cells in chilling-treated plants after heat acclimation (HA) and in heat-treated plants after cold acclimation (CA) were observed and compared using transmission electron microscopy. The results showed that slight injury appeared in the ultrastructure of mesophyll cells after either HA (38℃ for 10 h) or CA (8℃ for 2.5 d), but the tolerance to subsequent extreme temperature stress was remarkably improved by HA or CA pretreatment. The increases in membrane permeability and malondialdehyde concentration under chilling (0℃) or heat (45℃) stress were markedly inhibited by HA or CA pretreatment. The mesophyll cells of plants not pretreated with HA were markedly damaged following chilling stress. The chloroplasts appeared irregular in shape, the arrangement of the stroma lamellae was disordered, and no starch granules were present. The cristae of the mitochondria were disrupted and became empty. The nucleus became irregular in shape and the nuclear membrane was digested. In contrast, the mesophyll cells of HA-pretreated plants maintained an intact ultrastructure under chilling stress. The mesophyll cells of control plants were also severely damaged under heat stress. The chloroplast became round in shape, the stroma lamellae became swollen, and the contents of vacuoles formed clumps. In the case of mitochondria of control plants subjected to heat stress, the outer envelope was digested and the cristae were disrupted and became many small vesicles. Compared with cellular organelles in control plants, those in CA plant cells always maintained an integrated state during whole heat stress, except for the chloroplasts, which became round in shape after 10 h heat stress. From these data, we suggest that the stability of mesophyll cells under chilling stress can be increased by HA pretreatment. Similarly, CA pretreatment can protect chloroplasts, mitochondria, and the nucleus against subsequent heat stress; thus, the thermoresistance of grape seedlings was improved. The results obtained in the present study are the first, to our knowledge, to offered cytological evidence of cross-adaptation to temperature stresses in grape plants.  相似文献   

16.
Leaves from annual young grape plants (Vitis vinifera L. cv. Jingxiu) were used as experimentalmaterials. The ultrastructural characteristics of mesophyll cells in chilling-treated plants after heat acclima-tion (HA) and in heat-treated plants after cold acclimation (CA) were observed and compared using trans-mission electron microscopy. The results showed that slight injury appeared in the ultrastructure of meso-phyll cells after either HA (38℃ for 10 h) or CA (8℃ for 2.5 d), but the tolerance to subsequent extremetemperature stress was remarkably improved by HA or CA pretreatment. The increases in membrane perme-ability and malondialdehyde concentration under chilling (0℃) or heat (45℃) stress were markedly inhib-ited by HA or CA pretreatment. The mesophyll cells of plants not pretreated with HA were markedly dam-aged following chilling stress. The chloroplasts appeared irregular in shape, the arrangement of the stromalamellae was disordered, and no starch granules were present. The cristae of the mitochondria were dis-rupted and became empty. The nucleus became irregular in shape and the nuclear membrane was digested.In contrast, the mesophyll cells of HA-pretreated plants maintained an intact ultrastructure under chillingstress. The mesophyll cells of control plants were also severely damaged under heat stress. The chloroplastbecame round in shape, the stroma lamellae became swollen, and the contents of vacuoles formed clumps.In the case of mitochondria of control plants subjected to heat stress, the outer envelope was digested andthe cristae were disrupted and became many small vesicles. Compared with cellular organelles in controlplants, those in CA plant cells always maintained an integrated state during whole heat stress, except for thechloroplasts, which became round in shape after 10 h heat stress. From these data, we suggest that thestability of mesophyll cells under chilling stress can be increased by HA pretreatment. Similarly, CA pretreat-ment can protect chloroplasts, mitochondria, and the nucleus against subsequent heat stress; thus, thethermoresistance of grape seedlings was improved. The results obtained in the present study are the first,to our knowledge, to offered cytological evidence of cross-adaptation to temperature stresses in grapeplants.  相似文献   

17.
Two maize inbreds, CM7 and CM49, and CM7 × CM49, their F1 hybrid (which displayed significant heterosis), were examined with regard to response to exogenous gibberellin A3 (GA3), and in their ability to metabolize GA20, a native GA of maize. The leaf sheath elongation response to GA3 was far greater for the imbreds than for their hybrid. The inbreds also displayed significant elongation of the leaf blades in response to GA3, whereas the hybrid was unaffected. Promotion of cell division in the leaf sheath of CM7 and the hybrid was effected by GA3, but no promotion of cell elongation was observed in CM49, even though significant leaf sheath elongation occurred. Shoot dry weight of both inbreds was significantly increased by GA3, but response by the hybrid in this parameter was slight and variable. Root dry weight of CM7 was significantly increased by GA3, but was unchanged in CM49 and the hybrid. Thus, inbred shoot dry weight increases effected by GA3 were not at the expense of the root system. Rapid metabolism of [2,3-3H]GA20 occurred in all genotypes, although genotypic differences were observed. The hybrid had the highest rates of metabolism to GA glucosyl conjugate-like substances. Oxidative metabolism was also fastest in the hybrid, followed by CM7, and slowest in CM49, the slowest-growing inbred. Thus, rate of GA20 metabolism is under genetic control in normal (i.e. not dwarfed) maize genotypes. These results, taken together with previous reports that the hybrid has significantly enhanced levels of endogenous GA-like substances, suggest that GA play a role in the expression of heterosis in maize.  相似文献   

18.
Gibberellin A5 (GA5), a native GA of immature seeds of Pharbitis nil, was fed to Pharbitis nil cell suspension cultures as [C-l, 3H] GA5 (3.1 Ci/mmol), and its metabolism over a 48 hr period was investigated. Radioactivity in free GA metabolites was 13.1%, with 79.9% in GA glucosyl conjugate-like metabolites. Only 7.0% of the radioactivity remained as [3H] GA5. Tentative identifications were based on comparison with retention times of authentic free GAs and/or glucosyl conjugates after sequential chromatography on Si gel partition column → gradient-eluted C18 HPLC-radiocounting (RC) → isocratic-eluted C18 HPLC-RC, and showed that [3H] GA5 was converted to [3H] GA1 (2%), [3H] GA3 (4%), [3H] GA6 (2%), [3H] GA22 (1%) and their glucosyl conjugates, and also to [3H] GA8 glucoside, and [3H] GA5 glucosyl conjugates. The major conjugate-like substances were [3H] GA1 and [3H] GA3 glucosyl esters, at 15% and 34%, respectively, of the total extractable radioactivity.  相似文献   

19.
The hybrid Richter-110 (Vitis berlandieri x Vitis rupestris) (R-110) has the reputation of being a genotype strongly adapted to drought. A study was performed with plants of R-110 subjected to water withholding followed by re-watering. The goal was to analyze how stomatal conductance (g(s)) is regulated with respect to different physiological variables under water stress and recovery, as well as how water stress affects adjustments of water use efficiency (WUE) at the leaf level. Water stress induced a substantial stomatal closure and an increase in WUE, which persisted many days after re-watering. The g(s) during water stress was mainly related to the content of ABA in the xylem and partly related to plant hydraulic conductivity but not to leaf water potential. By contrast, low g(s) during re-watering did not correlate with ABA contents and was only related to a sustained decreased hydraulic conductivity. In addition to a complex physiological regulation of stomatal closure, g(s) and rate of transpiration (E) were strongly affected by leaf-to-air vapor pressure deficit (VPD) in a way dependent of the treatment. Interestingly, E increased with increasing VPD in control plants, but decreased with increasing VPD in severely stressed plants. All together, the fine stomatal regulation in R-110 resulted in very high WUE at the leaf level. This genotype is revealed to be very interesting for further studies on the physiological mechanisms leading to regulation of stomatal responsiveness and WUE in response to drought.  相似文献   

20.
The distribution frequency patterns of diameter of xylem vessels and percentage of total predicted axial conductances were studied in 190-day and 212-day-old main roots of grapevine (Vitis vinifera L. cv. Shiraz) grown under well-watered and stressed conditions. The protoxylem were the first to mature and were responsible for most of the theoretical conductance in root segments between the tip and 2.5 cm from the tip. Some large xylem vessels retained cross walls and protoplasm up to 22.5 cm from the tip. Statistical tests using the Kolmogorov-Smirnov two sample test showed that the pattern of distribution frequency of xylem vessels classified in different diameter classes varied with distance from the root tip. The distribution frequency of xylem vessels was similar in both well-watered and stressed plants from the tip up to 15 cm from the tip. At distances further from the tip the distribution frequency of xylem vessels of well-watered plants was significantly different from that of stressed plants, with the former having more larger vessels than the latter. The pattern of vessel distribution frequency was different from that of percent total axial conductance (Kh) predicted with fewer large vessels carrying most of the axial flow.  相似文献   

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