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1.
Kermode, A. R. and Bewley, J. D. 1985. The role of maturationdrying in the transition from seed development to germination.II. Post–germinative enzyme production and soluble proteinsynthetic pattern changes within the endosperm of Ricinus communisL. seeds.—J. exp. Bot. 36: 1916–1927. Immature seedsof Ricinus communis L. cv. Hale (castor bean) removed from thecapsule at 30 or 40 d after pollination (DAP) do not germinateunless first subjected to a desiccation treatment. This changefrom development to germination elicited by premature desiccationis also mirrored by a change, upon subsequent rehydration, inthe pattern of soluble protein synthesis within the endospermstorage tissue. Following rehydration of prematurely dried 30or 40 DAP seeds, soluble proteins characteristic of developmentcease to be synthesized after 5 h of imbibition, and those uniquelyassociated with germination and growth are then produced. Apattern of soluble storage protein breakdown comparable to thatfound in endosperms from mature seeds following imbibition isalso observed. In contrast, hydration of 40 DAP seeds immediatelyfollowing detachment from the mother plant results in a continuationof the developmental pattern of protein synthesis. Prematuredesiccation at 40 DAP elicits the production within the endospermof enzymes involved in protein reserve breakdown (leucyl ß–naphthylamidase;LeuNAase) and lipid utilization (isocitrate lyase; ICL) to levelscomparable to those observed in mature–hydrated endosperms.It is proposed that drying plays a role in redirecting metabolismfrom a developmental to a germinative mode; it also appearsto be a prerequisite for the induction of hydrolytic enzymesessential to the post–germinative (growth) phase of seedlingdevelopment. Key words: Desiccation-tolerance, germinability, seed development, castor bean  相似文献   

2.
Drying of immature seeds of Ricinus communis L. cv. Hale (castorbean) during the desiccation-tolerant phase of development causesthem to germinate upon subsequent rehydration. This desiccation-inducedswitch from development to germination is also mirrored by achange in the pattern of soluble and insoluble protein synthesiswithin the cotyledons of the castor bean. Following rehydrationof seeds prematurely dried at 40 d after pollination (DAP),cotyledonary proteins characteristic of development (e.g. storageproteins) are no longer synthesized; hydrolytic processes resultingin their degradation commence (after 12 h) in a manner similarto that observed following imbibition of the mature seed. Apattern of protein synthesis recognizable as germination/growth-associatedoccurs; premature drying has elicited a redirection in metabolismfrom a developmental to a germinative mode. Desiccation is alsorequired for the induction (within cotyledons of 35 DAP seeds)of enzymes involved in protein reserve breakdown (leucyl ß-naphthylamidase;LeuNAase) and lipid utilization (isocitrate lyase; ICL), anevent intimately associated with the post-germinative (growth)phase of seedling development. Thus, at a desiccation-tolerantstage of development, premature drying results in the suppressionof the developmental metabolic programme and a permanent switching-onof the germination/growth metabolic programme. Key words: Desiccation, metabolism, seed development, seed germination, castor bean, cotyledons  相似文献   

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Proteins in the soluble and insoluble fractions, extracted frommature castor bean cv. Hale seed cotyledons, differ quantitativelyand qualitatively from their counterparts extracted from theendosperm. The soluble fraction contains no glycoproteins, andthe lectins RCA1 and ricin D are absent. While the insolubleproteins are electrophoretically and immunologically similarto those in the endosperm, they do not form the 100 kD subunitdimers which characterize some of the endosperm insoluble crystalloidproteins. Rapid rates of deposition of all of the soluble andinsoluble proteins present in the mature seed cotyledons commences30–35 d after pollination (DAP) and continues until 45DAP. These proteins are mobilized rapidly beginning 1–2d after seed imbibition and this coincides with an increasein specific activity, in the cotyledons, of two aminopeptidasesand a carboxypeptidase. The soluble and insoluble proteins inthe cotyledons of the mature seed probably function as storageproteins and support the growth of the germinated seed priorto the mobilization of the major protein storage reserves ofthe endosperm. Key words: Ricinus communis, Castor bean, Hale cultivar, Cotyledon, Storage protein, Seed development, Seed germination  相似文献   

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Hydrolysis of the insoluble crystalloid storage proteins ofcastor bean endosperm during germination released buffer-solublepolypeptides with molecular weights in the presence of sodiumdodecyl sulphate of 30000–40000. These polypeptides appearto be dimers since the addition of 2-mercaptoethanol decreasestheir molecular weights to 15000–22000. Hydrolysis ofthe crystalloid proteins was detected 12–18 h after seedimbibition (HAI), which is before the completion of germination;maximum rates were attained at 30 HAI. During this period, parallelincreases in free amino acids were observed. Hydrolysis of thecrystalloid proteins during early germination was insensitiveto cycloheximide treatment and therefore did not require newlysynthesized proteases. Hydrolysis was effected by proteaseswhich were made in an inactive form during seed developmentand activated upon seed imbibition. Key words: Castor bean, crystalloid storage protein hydrolysis, seed germination, endosperm  相似文献   

8.
During mid-development (25–40 d after pollination: DAP)of the castor bean seed the amount of abscisic acid (ABA) increasesin both the endosperm and the embryo, declining substantiallythereafter until there is little present in the mature dry (60DAP) seed. Premature desiccation of the seed at 35 DAP alsoleads to a major decline in ABA within the embryo and endosperm.Partial water loss from the seed at 35 DAP which, like naturaland premature desiccation, leads to subsequent germination uponreturn of the seed to full hydration, causes a much smallerdecline in ABA levels. In contrast, ABA declines substantiallyin the non-dried (hydrated) control at 35 DAP, but the seedsdo not germinate. Hence, a clear negative correlation betweenABA content and germinability is not observed. Both drying,whether natural or imposed prematurely, and partial drying decreasethe sensitivity of the isolated embryo to exogenous ABA by about10-fold. The protein synthetic response of the castor bean embryo exposedto 0.1 mol m–3 ABA following premature desiccation exhibitssome similarity to the response of the non-dried developingembryo—in both cases the synthesis of some developmentalproteins is enhanced by ABA, and germination is suppressed.Germination of mature seeds is also suppressed by 0.1 mol m–3ABA, but the same developmental proteins are not synthesized.In the cotyledons of prematurely-desiccated seed, some proteinsare hydrolysed upon imbibition in 0.1 mol m–3 ABA, a phenomenonthat occurs also in the cotyledons of similarly treated matureembryos, but not in developing non-dried embryos. Hence theembryo exhibits an ‘intermediate’ response uponrehydration in 0.1 mol m–3 ABA following premature desiccation;viz. some of the responses are developmental and some germinative.Following natural or imposed drying, the isolated embryo becomesrelatively insensitive to 0.01 mol m–3 ABA: germinationis elicited and post-germinative reserve breakdown occurs inthe radicle and cotyledons. The reduced sensitivity of the embryoto ABA as a consequence of desiccation may be an important factorin eliciting the switch to germination and growth within thewhole seed. Key words: Abscisic acid, desiccation, astor bean endosperm, seed development, germination, protein synthesis, isolated embryos, hormone sensitivity  相似文献   

9.
Misra, S. and Bewley, J. D. 1986. Desiccation of Phaseolus vulgansseeds during and following germination, and its effect uponthe translatable mRNA population of the seed axes.—J.exp. BoL 37: 364–374. After imbibition and germination, seeds of P. vulgaris passfrom a stage where they are insensitive to desiccation to astage where they are sensitive. Desiccation of seeds duringthe sensitive stage results in an almost total impairment ofprotein synthesis upon subsequent rehydration. Seeds desiccatedduring the desiccation-tolerant stage, however, resume proteinsynthesis at almost control levels. The protein patterns obtained following in Vitro translationof bulk RNA from fresh imbibed, desiccated, and desiccated-rehydratedseed axes were qualitatively similar at 5 HAI (the desiccation-tolerant stage). The drying treatment resulted in increasedintensity of extant proteins at 5 and 12 HAI. At 12 HAI (thetransition stage between the desiccation-tolerant and desiccation-intolerantphases) desiccation and subsequent rehydration triggered synthesisof a unique set of proteins-the rehydration proteins. At 20HAI (the desiccation-intolerant stage) desiccation resultedin an overall decline in the intensity of proteins synthesizedin vitro. Also the rehydration proteins were not synthesizedin response to a drying and rehydration treatment at this time. Key words: Seed germination, desiccation, mRNA, in vitro translation, Phaseolus vulgaris  相似文献   

10.
Kennode, A. R, and Bewley, J. D. 1988. The role of maturationdrying in the transition from seed development to germination.V. Responses of the immature castor bean embryo to isolationfrom the whole seed; a comparison with premature desiccation.—J.exp. Bot. 39: 487–497. Desiccation is an absolute requirement for germination and post-germinativegrowth of whole seeds of the castor bean, whether desiccationis imposed prematurely during development, at 35 d after pollination(DAP) or occurs naturally during late maturation (50–60DAP). Desiccation also plays a direct role in the inductionof post-germinative enzyme synthesis in the cotyledons of embryosin the intact seed; this event is not simply due to the presenceof a growing axis. Isolation of embryos from the developingcastor bean seed at 35 DAP results in both germination and growth,despite the absence of a desiccation event. We have comparedthe metabolic consequences of premature drying of whole seeds(35 DAP) and isolation of the developing 35 DAP embryos. Inboth cases, hydrolytic events involved in the mobilization ofstored protein reserves proceed in a similar manner and mirrorthose events occurring within germinated mature seeds. Thereare differences, however, for post-germinative enzyme (LeuNAaseand isocitrate lyase) production occurs to a lesser extent innon-dried isolated embryos than in those from prematurely dried(35 DAP) whole seeds, or from mature dry (whole) seeds. Desiccationof the 35 DAP whole seed does not alter the subsequent responseof the embryo upon isolation. Thus, while drying does not affectthe metabolism of isolated embryos, it has a profound effecton that of embryos within the intact seed. Tissues surroundingthe embryo in the developing intact seed (viz. the endosperm)maintain its metabolism in a developmental mode and inhibitgermination. This effect of the surrounding tissues can onlybe overcome by drying or by their removal. Key words: Metabolism, isolation, desiccation, embryo, endosperm, castor bean, development, germination  相似文献   

11.
Kermode, A. R. and Bewley, J. D. 1985. The role of maturationdrying in the transition from seed development to germination.1. Acquisition of desiccation–tolerance and germinabilityduring development of Ricinus communis L. seeds.—J. exp.Bot. 36: 1906–1915. Seeds of Ricinus communis L. cv. Hale(castor bean) undergo a transition from desiccation–intoleranceto desiccation–tolerance approximately midway throughtheir development. Tolerance of slow desiccation is gained overonly a few days of development (between 20 and 25 d) and isachieved well before the completion of major developmental events,such as reserve deposition and the onset of normal maturationdrying. A tolerance of very rapid water loss brought about bydrying over silica gel is not acquired by this seed until nearmaturity. Coincident with the acquisition of tolerance to slowdesiccation the seeds gain the capacity to germinate upon subsequentrehydration. Germinability and capacity for normal post–germinativegrowth during the tolerant phase are not fully expressed unlessthe seed is dried at an optimal rate, which is dependent uponthe developmental stage of the seed. Drying presumably actsto terminate developmental processes and to initiate those metabolicprocesses necessary to prepare the seed for germination andgrowth. Key words: Desiccation-tolerance, germinability, seed development, castor bean  相似文献   

12.
Seed water content is high during early development of tomato seeds (10–30 d after pollination (DAP)), declines at 35 DAP, then increases slightly during fruit ripening (following 50 DAP). The seed does not undergo maturation drying. Protein content during seed development peaks at 35 DAP in the embryo, while in the endosperm it exhibits a triphasic accumulation pattern. Peaks in endosperm protein deposition correspond to changes in endosperm morphology (i.e. formation of the hard endosperm) and are largely the consequence of increases in storage proteins. Storage-protein deposition commences at 20 DAP in the embryo and endosperm; both tissues accumulate identical proteins. Embryo maturation is complete by 40 DAP, when maximum embryo protein content, size and seed dry weight are attained. Seeds are tolerant of premature drying (fast and slow drying) from 40 DAP.Thirty-and 35-DAP seeds when removed from the fruit tissue and imbibed on water, complete germination by 120 h after isolation. Only seeds which have developed to 35 DAP produce viable seedlings. The inability of isolated 30-DAP seed to form viable seedlings appears to be related to a lack of stored nutrients, since the germinability of excised embryos (20 DAP and onwards) placed on Murashige and Skoog (1962, Physiol. Plant. 15, 473–497) medium is high. The switch from a developmental to germinative mode in the excised 30- and 35-DAP imbibed seeds is reflected in the pattern of in-vivo protein synthesis. Developmental and germinative proteins are present in the embryo and endosperm of the 30- and 35-DAP seeds 12 h after their isolation from the fruit. The mature seed (60 DAP) exhibits germinative protein synthesis from the earliest time of imbibition. The fruit environment prevents precocious germination of developing seeds, since the switch from development to germination requires only their removal from the fruit tissue.Abbreviations DAP days after pollination - kDa kilodaltons - SP1-4 storage proteins 1–4 - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis - HASI hours after seed isolation - MS medium Murashige and Skoog (1962) medium This work is supported by National Science and Engineering Research Council of Canada grant A2210 to J.D.B.  相似文献   

13.
Soybean seeds [Glycine max (L.) Merr.] synthesize de novo andaccumulate several non-storage, soluble polypeptides duringnatural and precocious seed maturation. These polypeptides havepreviously been coined ‘maturation polypeptides’.The objective of this study was to determine the fate of maturationpolypeptides in naturally and precociously matured soybean seedsduring rehydration, germination, and seedling growth. Developingsoybean seeds harvested 35 d after flowering (mid-development)were precociously matured through controlled dehydration, whereasnaturally matured soybean seeds were harvested directly fromthe plant. Seeds were rehydrated with water for various timesbetween 5 and 120 h. Total soluble proteins and proteins radio-labelledin vivo were extracted from the cotyledons and embryonic axesof precociously and naturally matured and rehydrated seed tissuesand analyzed by one-dimensional PAGE and fluorography. The resultsindicated that three of the maturation polypeptides (21, 31and 128 kDa) that had accumulated in the maturing seeds (maturationpolypeptides) continued to be synthesized during early stagesof seed rehydration and germination (5–30 h after imbibition).However, the progression from seed germination into seedlinggrowth (between 30 and 72 h after imbibition) was marked bythe cessation of synthesis of the maturation polypeptides followedby the hydrolysis of storage polypeptides that had been synthesizedand accumulated during seed development. This implied a drasticredirection in seed metabolism for the precociously maturedseeds as these seeds, if not matured early, would have continuedto synthesize storage protein reserves. Glycine max (L.) Merr, soybean, cotyledons, maturation, germination/seedling growth  相似文献   

14.
Protein bodies within the endosperm of castor bean (Ricinus communis L. cv. Hale) seeds arise from numerous small vacuoles which progressively become filled with storage protein, of which the crystalloid proteins make up approximately 70%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) shows that the crystalloids are a family of at least four proteins which reduce to two complementary groups after 2-mercaptoethanol treatment. The matrix, which comprises the remainder, has two major components, the soluble albumins and the lectins. The lectins are the only glycoproteins within the mature protein body. Both cytochemical staining and SDS-PAGE indicate that the synthesis of the crystalloid and the majority of matrix proteins begins some 20 days after pollination. Additionally, the crystalloid proteins are synthesized concurrently, whereas there is temporal variation in the synthesis of matrix proteins.  相似文献   

15.
Drying of seeds at certain stages prior to maturation, i.e.premature desiccation, will terminate synthetic events uniqueto development, for example, storage protein synthesis, andinitiate processes associated with germination. In this studywe have investigated the role of desiccation in the expressionof a storage protein gene, ß-phaseolin, to determineif such a developmentally-regulated gene remains sensitive todrying when controlled by a promoter that has no known sensitivityto this treatment. We compared, in transgenic tobacco seeds,the effects of maturation and premature drying on the expressionof a full ß-phaseolin gene, and ß-phaseolingenes driven by a cauliflower mosaic virus 35S promoter withor without an alfalfa mosaic virus (AMV) 5' untranslated leadersequence. The results indicate that the ß-phaseolinpromoter is directly down-regulated by desiccation during maturationand, although activated during the drying phase of a prematuredesiccation event, it is not active upon rehydration or imbibition.The 35S promoter is down-regulated also by both maturation dryingand premature desiccation but unlike the ß-phaseolinpromoter it is reactivated upon rehydration or imbibition. Key words: Desiccation, ß-phaseolin, gene regulation, Phoseolus vulgaris, seed development  相似文献   

16.
A spectrophotometric assay was devised to characterize the asparaginyl (Asn) endopeptidase activity from the endosperm of castor oil seeds. (Ricinus communis L. var. Baker 296). The assay measures the release of p-nitroaniline from the hydrolysis of benzoyl-l-Asn-p-nitroanilide. Assay sensitivity was improved through diazotization of the reaction product with N(]-napthy])-ethylenediamine dihydrochloride: diazotized p-nitroaniline was determined spectrophotometrically at 548 nm (?548= 1.64 × 10?1M?1 cm?2). By using this assay. Asn endopeptidase activity was detected in endosperm extracts of developing, mature and germinating castor seeds. Comparison of the Asn endopeptidase activities of developing and germinating castor endosperms revealed that they: 1) have identical pH-activity profiles with optimal activity occuring at pH 5.4: 2) are heat-labile proteins displaying comparable thermal stability profiles, and 3) are activated and inhibited by dithiothreitol and thiol modifying reagents, respectively. Thus, the Asn endopeptidases of developing and germinating castor seeds are very similar, if not identical, cysteine proteases. The most significant increase in the activity of endosperm Asn endopeptidase occurs during the full coryledon to maturation stage of seed development, this period coincides with the most active phase of reserve protein accumulation by ripening castor oil seeds. Asn endopeptidase activity of fully mature (dry) castor seeds was about 2-fold lower than that of muturation stage ripening castor oil seed. Asn endopeptidase activity showed a slight reduction over the inicial 2-day period following seed imbibition, and then rapidly decreased over the next several days of germination. The results are compatible with the proposal that Asn endopeptidase functions both to process storage preproteins following their import into protein bodies of developing seeds, as well as to participate in the mobilization of storage proteins during the early phase of seed germination.  相似文献   

17.
Cytochemical staining of sections prepared for light microscopy,electron microscope sections, and sodium dodecyl sulphate-polyacrylamidegel electrophoresis reveal that, following imbibition, storageproteins are mobilized from the protein bodies of the endospermof castor bean (Ricinus communis L. cv. Hale). This is accompaniedby fusion of protein bodies to form a central vacuole, beforeall the protein is hydrolyzed. Mobilization of the US crystalloidprotein complex and of the 2S albumin fraction commences 2 dafter imbibition and is completed within 2 d. This loss of proteinis accompanied by an increase in activity of three proteolyticenzymes, one carboxypeptidase and two -SH-dependent aminopeptidases.In contrast to the 11S and 2S protein fractions the lectins,located within the protein body, are mobilized only slowly andare present after the other proteins have been completely brokendown. Hence lectins may have a role other than as storage proteins. Key words: Castor bean, Protein breakdown, Storage protein, Lectin, Vacuolation, Seed germination  相似文献   

18.
Freshly harvested, developing kernels of maize (Zea mays L.)do not germinate up to 77 d after pollination, but can be inducedto do so by fluridone, premature desiccation, and isolationof the developing embryo. The pattern of protein synthesis indeveloping maize embryos is distinct from that during germinationand subsequent seedling growth. Premature desiccation at 35DAP elicits a pattern of protein synthesis upon rehydrationwhich is similar to that in germinated embryos from mature drykernels. Fluridone-induced viviparous germination is accompaniedby changes in the synthesis of some proteins to a post-germinativepattern, but some developmental proteins continue to be synthesized.Embryos isolated from developing kernels at 35 DAP germinatewhen incubated on water; they also produce some developmentalproteins during germination. Kernels from developing cobs at35 DAP which are detached from the mother plant and maintainedin an atmosphere of high relative humidity (moist controls)do not germinate, but neither do they continue a clearly definedpattern of either developmental or germinative protein synthesis.Drying is thus critical to effect a clear transition of proteinsynthesis from a developmental to a germinative mode in maizeembryos. Abscisic acid within the developing embryos is reduced by fluridone,but to a lesser extent by premature drying or maturation drying.Changes in sensitivity to abscisic acid by the developing embryomay be as, or more, important in permitting germination, andthe attendant synthesis of proteins, than changes in abscisicacid content. Key words: Maize (Zea mays L.), germination, vivipary, desiccation, abscisic acid  相似文献   

19.
Monospecific polyclonal antibodies against maize leaf phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) were utilized to examine the subunit composition and developmental profile of endosperm PEPC in developing and germinating castor oil seeds (Ricinus communis L. cv Baker 296). PEPC from developing endosperm consists of a single type of 100-kilodalton subunit, whereas the enzyme from 2- to 5-day germinated endosperm appears to contain equal proportions of immunologically related 103- and 108-kilodalton subunits. The maximal activity of PEPC in developing endosperms (2.67 micromoles oxaloacetate produced per minute per gram fresh weight) is approximately 20-fold and threefold greater than that of fully mature (dry seed) and germinating endosperms, respectively. The most significant increase in the activity and concentration of endosperm PEPC occurs during the middle cotyledon to full cotyledon stage of seed development; this period coincides with the most active phase of storage oil accumulation by ripening castor oil seeds. The data are compatible with the recent proposal (RG Smith, DA Gauthier, DT Dennis, DH Turpin [1992] Plant Physiol 1233-1238) that PEPC plays a fundamental role in vivo in the cytosolic production of an important substrate (malate) for fatty acid biosynthesis by developing castor oil seed leucoplasts. Immediately following seed imbibition, PEPC activity and concentration increase in parallel, with the greatest levels attained by the third day of germination. It is suggested that during this early phase of seed germination PEPC has a critical function to build up cellular dicarboxylic acid pools required to initiate significant activities of both the tricarboxylic acid and glyoxylate cycles.  相似文献   

20.
The embryo axis is required for the rapid breakdown of the crystalloid,albumin and lectin protein storage reserve in the endospermof castor bean (Ricinus communis L. cv. Hale) seeds, and forthe attainment of high specific activities of several endospermicproteolytic enzymes: one carboxy-peptidase and two -SH- dependentaminopeptidases. The embryo axis must be present to initiatestorage protein breakdown but it is not required to maintainthis process. We suggest that the embryo axis controls storageprotein breakdown through the release of promoters, which canbe replaced by gibberellins. Storage protein breakdown is notinfluenced by source-sink effects. However, the endosperm becomessensitive to gibberellin only after an imbibition period forup to 24 h. Key words: Castor bean, Protein breakdown, Storage protein, Embryo control, Gibberellin, Seed germination  相似文献   

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