The Role of Maturation Drying in the Transition from Seed Development to Germination: III. INSOLUBLE PROTEIN SYNTHETIC PATTERN CHANGES WITHIN THE ENDOSPERM OF Ricinus communis L. SEEDS

Kermode, A. R., Gifford, D. J. and Bewley, J. D. 1985. The roleof maturation drying in the transition from seed developmentto germination. III. Insoluble protein synthetic pattern changeswithin the endosperm of Ricinus communis L. seeds.—J.exp. Bot. 36: 1928–1936. Immature seeds of Ricinus communisL. cv. Hale (castor bean) removed from the capsule at 30 or40 days after pollination (DAP) can be induced to germinateby being subjected to drying. This desiccation–inducedswitch from development to germination is mirrored by a change,upon subsequent rehydration, in the pattern of insoluble proteinsynthesis within the endosperm storage tissue. During normaldevelopment from 25–40 DAP there is rapid synthesis ofthe insoluble (11S) crystalloid storage protein. At later stagesof development (45 and 50 DAP), crystalloid protein synthesisdeclines markedly and synthesis of new insoluble proteins commences.Following premature drying at 30 or 40 DAP, the pattern of insolubleprotein synthesis upon rehydration is virtually identical tothat following imbibition of the mature seed. Proteins synthesizedduring normal late development (at 45 and 50 DAP) are producedup to 48 h after imbibition; a subsequent change in the patternof insoluble protein synthesis occurs between 48 and 72 h. Thus,in contrast to the rapid switch in the pattern of soluble proteinsynthesis induced by drying, insoluble protein syntheses withinthe endosperm are redirected towards those uniquely associatedwith a germination/growth programme only after a considerabledelay following mature seed imbibition, or following rehydrationof the prematurely dried seed. Nevertheless, these results supportour contention that drying plays a role in the suppression ofthe developmental metabolic programme and in the permanent inductionof a germination/growth programme. Key words: Desiccation, crystalloid storage proteins, castor bean, seed development, seed germination     

The Role of Maturation Drying in the Transition from Seed Development to Germination: I. ACQUISITION OF DESICCATION-TOLERANCE AND GERMINABILITY DURING DEVELOPMENT OF Ricinus communis L. SEEDS

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     

The Role of Maturation Drying in the Transition from Seed Development to Germination: II. POST-GERMINATIVE ENZYME PRODUCTION AND SOLUBLE PROTEIN SYNTHETIC PATTERN CHANGES WITHIN THE ENDOSPERM OF Ricinus communis L. SEEDS

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     

Desiccation of Phaseolus vulgaris Seeds During and Following Germination, and its Effect Upon the Translatable mRNA Population of the Seed Axes

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     

Stable Delivery of a Canavalin Promoter-{beta}-Glucuronidase Gene Fusion into French Bean by Particle Bombardment

Shoot apices of French bean (Phaseolus vulgaris L. cv. Goldstar)seeds were bombarded with gold particles coated with plasmidpSAG-734 carrying a chimeric gene that encodes ß-glucuronidaseunder the control of the canavalin gene promoter. The organ-and maturation stage-specific expression of the construct wasobserved histochemically in seeds of transgenic plants (Received July 10, 1996; Accepted October 5, 1996)     

Seed development and maturation in Phaseolus vulgaris I. Ability to germinate and to tolerate desiccation

The onset and development of both the ability to germinate andto tolerate rapid enforced desiccation were investigated duringthe development and maturation of seeds of bean (Phaseolus vulgahsL.) at different temperatures and also after different slow-dryingtreatments. The onset of germinability occurred when seeds wereless than half-filled in the absence of both a post-ovule abscissionprogramme and water loss from the seeds. Maximum ability togerminate normally and maximum tolerance to rapid enforced desiccationto 14–16% moisture content did not occur until 2–23d and 6–23 d after mass maturity (end of the seed-fillingperiod), respectively. The slow-drying of immature seeds for7 d ex planta before rapid enforced desiccation increased theability to germinate and stimulated the onset of desicationtolerance. Holding seeds moist for 7 d (during which time moisturecontent declined by <5%) had similar effects, but seed germinationafter rapid enforced desiccation was consistently greater inseeds first dried slowly than held moist. Comparisons betweenseeds less than half-filled dried slowly ex planta and fullseeds undergoing maturation drying in planta showed that a similar(slow) rate of water loss over a 7 d period had a similar effecton the subsequent ability of seeds to tolerate rapid enforceddesiccation. Thus, neither a post-ovule abscission programmenor loss of water were required for the onset of the abilityto germinate in developing bean seeds, but both were requiredfor the development of the ability to germinate and resistanceto solute leakage, when rehydrated, after rapid enforced desiccation. Key words: Bean, Phaseolus vulgaris L., seed germination, seed development, desiccation tolerance     

Seed development and maturation in Phaseolus vulgaris II. Post-harvest longevity in air-dry storage

Changes in seed quality were monitored during the developmentand maturation of seeds of bean (Phaseolus vulgaris L.) in differentenvironments by determining the subsequent survival of seedsin airdry storage. The seed survival curves (percentage normalgermination plotted against period of storage) conformed tonegative cumulative normal distributions, and the same seedstorage environment (40 C with 14% moisture content) providedthe same estimate of the standard deviation of the frequencydistribution of seed deaths in time for all the seed lots harvestedat different times during development in the different environments(21.3 d and 20.9 d in 1993 and 1994, respectively). The potentiallongevity of developing seeds (quantified by the value of theseed lot constant of the seed viablity equation) continued toincrease after mass maturity (end of the seed-filling phase).Maximum potential longevity was attained 23 d (30/24 C, 1993),13 d (30/24 C, 1994), or 34 d (27/21 C, 1994) after mass maturity,by which time seeds had dried naturally to 17% (30/24 C, 1993),18% (30/24 C, 1994), and 16% (27/21 C, 1994) moisture content.Seed quality was greater in the cooler temperature seed productionregime because of an increased duration of seed quality improvement.The results show that maximum seed quality is not attained untilsome time after the end of the seed-filling phase in bean, andconfirm that the slow desiccation that occurs after ovule abscissionduring natural maturation drying is beneficial to the developmentof seed quality in bean. Key words: Bean, Phaseolus vulgaris L., seed development, seed longevity, seed storage     

Stereospecificity of hydrogen transfer by pyridine nucleotide-dependent enoyl reductases in fatty acid synthesis: Studies with enzymes obtained from developing castor bean seeds and Chlorella vulgaris

The mechanism of hydrogen incorporation into fatty acids wasinvestigated with fatty acid synthetase systems from developingcastor bean seeds and Chlorella vulgaris. Fatty acids synthesizedin the presence of D₂O or stereospecifically deuterium-labeledNADPH or NADH were isolated and analyzed by mass spectrometryto examine the localization of deuterium atoms in the molecule.The stereospecificities of ß-ketoacyl-acyl carrierprotein (ACP) reductase and enoyl-ACP reductase for reducedpyridine nucleotide were determined with acetoacetyl-ACP andcrotonyl-ACP as substrates. The products were also analyzedby gas chromatography-mass spectrometry. The following resultswere obtained:

1. ß-Ketoacyl-ACP reductases from both castor bean seedsand C. vulgaris used the B-side hydrogen of NADPH.
2. Enoyl-ACPreductase from C. vulgaris required NADH for the activity.
3. Enoyl-ACPreductase from castor bean seeds used the A-side hydrogenofNADPH, whereas that from C. vulgaris used the B-side hydrogenof NADH.
4. When stearate was synthesized with the crude fattyacid synthetasefraction from castor bean seeds, hydrogen atomsfrom water werefound on the even-numbered methylene carbonatoms (two hydrogenatoms per carbon atom) and some were foundon the odd-numberedmethylene carbon atoms. Hydrogen atoms fromthe B-side of NADPHwere found on the odd-numbered methylenecarbon atoms (one hydrogenatom per carbon atom). Hydrogen atomsfrom the A-side of NADPHwere also found on the odd-numberedmethylene carbon atoms,but the number of incorporated hydrogenatoms was less thanexpected.

(Received October 17, 1979; )     

The Role of Maturation Drying in the Transition from Seed Development to Germination: IV. PROTEIN SYNTHESIS AND ENZYME ACTIVITY CHANGES WITHIN THE COTYLEDONS OF RICINUS COMMUNIS L. SEEDS

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     

Comparative Fungitoxic and Plant Growth Regulating Properties of Triazole Derivatives

The triazol derivatives, S-3307, S-3308, triadimefon, triadimenoland paclobutrazol are recommended for use as either fungicidesor plant growth regulators. However, in varying degrees theyexhibit both properties. In a comparative study with 5 differentfungi, S-3308 followed by S-3307 were the most fungitoxic (invitro) whereas S-3307 and paclobutrazol were the most activeplant growth retardants in bean (Phaseolus vulgaris L.) andKentucky bluegrass (Poa pratensis L.). (Received September 24, 1985; Accepted November 26, 1985)     

Changes in Lipids of Bean Seeds (Phaseolus vulgaris) and Corn Caryopses (Zea mays) Aged in Contrasting Environments

Seeds of French bean (Phaseolus vulgaris L. cv. The Prince)and caryopses of sweet corn (maize; Zea mays L.F1 hybrid Firstof All) were stored in environments of 79% relative humidityand 25 °C, 80% r.h. and 40 °C or 100% r.h. and 45 °C,giving ageing (loss of gcrminability and vigour) over periodsof months, weeks or days, respectively. The relationship betweenchanges in lipids and changes in germinability or vigour wasunaffected in general by the speed of ageing. In the corn caryopsesthere was no evidence of hydrolysis of phospholipids or peroxidationof fatty acids during ageing. In the bean seeds phosphatidicacid increased during the ageing period and phosphatyl cholinedeclined. The percentage of fatty acids as hnolenic acid initiallyfell during bean ageing, but in the slower ageing conditionsit rose again as germination reached zero. In bean, the presenceof phosphatidic acid could be a sensitive indictator of lossof vigour, but relative proportions of the different fatty acidswould be a misleading indicator of quality. Rapid artificialageing may be an adequate model, in some species, of ageingat moderate speeds and of ageing under some ambient conditions. French bean (Phaseolus vulgaris L. cv. The Prince), sweet corn (maize, Zea mays L., Fl hybrid First of All), seed ageing, phospholipids, fatty acids     

Regeneration of Fertile Plants from Callus in Phaseolus polyanthus Greenman (Year Bean)

For the first time, plant regeneration of several domesticatedgenotypes of Phaseolus polyanthus Greenman (year bean) has beenachieved. Thidiazuron in combination with indole-3-acetic acidwas used to induce morphogenic, green nodular callus from explantsthat had been obtained either from greenhouse-grown plants orfrom in vitro -germinated seeds. Of the six genotypes of P.polyanthus tested, five produced shoots in vitro. Regeneratedshoots that formed roots in vitro were established in the greenhouse,whereas non-rooted shoots could be established in vitro by grafting.Morphologically normal progeny plants were obtained from thegreenhouse-established regenerants. However, by using the sameprocedure, no regeneration response was observed in two domesticatedand two wild genotypes of P. coccineus L. (runner bean). Thisprotocol should help achieve Agrobacterium - or particle bombardment-mediatedgenetic transformation to improve this important food legume.Copyright 2001 Annals of Botany Company Callus, genetic transformation, grafting, legumes, organogenesis, Phaseolus coccineus L., Phaseolus polyanthus Greenman, Phaseolus vulgaris L     

Internodal Extension in the First Trifoliate Leaf Axillary Bud of Phaseolus vulgaris L. following Shoot Decapitation

Phaseolus vulgaris L. decapitated at the third internode showedaccelerated growth of the uppermost axillary bud remaining onthe stem (the first trifoliate axillary bud) after a lag periodof 3–5 h Much of the initial growth increment could beattributed to cell expansion Phaseolus vulgaris L, dwarf bean, correlative inhibition, cell expansion     

Deoxyribonucleic Acid, Ribonucleic Acid, Protein and Uncombined Amino Acid Content of Legume Seeds During Embryogeny

The nucleic acid, protein and uncombined amino acid contentof seeds of soya-bean (Glycine max L. Merr.), garden pea (Pisumsativum L.), kidney bean (Phaseolus vulgaris L.) and peanut(Arachis hypogaea L.) were measured at various times duringseed formation in an effort to understand why the soya-beanhas nearly twice as much protein as the other legume seeds.In all these species the concentration of deoxyribonucleic acid,ribonucleic acid and uncombined amino acids decreased duringseed formation. The protein level of kidney bean was relativelyconstant during development whereas the protein levels of pea,peanut and soya-bean increased during development. The proteincontent of the soya-bean increased throughout development whereasthe protein increase in peanut took place early and that inpea took place later in development. The ratio of protein toribonucleic acid was highest in peanut, less in soya-bean, andlowest in pea and kidney bean. Similarly, the ratio of proteinto deoxyribonucleic acid was higher in kidney bean than in soya-bean.Soya-beans had a lower amino acid content than any of the otherseeds at all stages of development. These results indicate thatneither total deoxyribonucleic acid, ribonucleic acid nor uncombinedamino acid content is responsible for the higher protein contentof soya-beans.     

Some Effects of Sucrose Concentration on Primordium Development in Excised Primary Roots

The effects of varying the amount of sucrose used to supplementthe culture medium maintaining the growth of excised roots ofPisum sativum L., Vicia faba L., Zea mays L. and Phaseolus vulgarisL., on the rates of primordium initiation and subsequent emergenceas lateral roots and on the duration of the interval betweenprimordium inception and emergence as a secondary root throughthe tissues of the primary have been investigated. Variation in the exogenous concentration of sucrose from 0.5to 8 per cent had little effect on the rate of primordium inceptionin Pisum and Vicia and the rates never reached the values obtainedfor the roots of the corresponding intact plants. Moreover,over the 6 day culture period lateral root emergence did notoccur in any of the excised roots of these two species. In contrast,each of the aspects of primordium development examined in theexcised roots of Zea and Phaseolus was markedly affected bythe amount of sucrose used to supplement the culture medium.In addition, in the presence of about 6 per cent sucrose, primordiumdevelopment in these cultured roots was very similar to thatin roots of the corresponding intact plants. The results indicate either that some factor necessary for primordiumdevelopment is present in adequate amounts in excised rootsof Zea and Phaseolus, but not in those of Pisum and Vicia, orthat the factors controlling such development are differentin the former and latter two species. Vicia faba L., Pisum sativum L., Zea mays L., Phaseolus vulgaris L., broad bean, garden pea, maize, dwarf bean, primordium development, sucrose concentration, cultured roots     

Sodium Recirculation and Loss from Phaseolus vulgaris L.

In a split-root experiment, ²²Na was supplied to Phaseolus vulgarisL. roots emerging from the stem, 2.5 cm above the main roots.Sodium exported from these upper roots was translocated a shortdistance upward in the stem and downward to the main roots.Most of the ²²Na arriving in the main roots was lost to themedium. Sodium loss from P. vulgaris roots into KCI or NaCl was similarand was not affected by oligomycin. The results confirm a previous hypothesis regarding the mechanismof sodium exclusion from the tops of sodium non-accumulatorplants. Phaseolus vulgaris L., bean, sodium transport     

Lipoxygenase, Hydroperoxide Lyase and Volatile C6-Aldehyde Formation from C18-Fatty Acids during Development of Phaseolus vulgaris L.

Kidney bean plants (Phaseolus vulgaris) were found to have thecapability to produce C₆-aldehydes (hexanal and hexenals) fromlinoleic and linolenic acids. The various organs tested hadlipoxygenase and hydroperoxide lyase activities responsiblefor the C₆-aldehyde formation. Young leaves showed relativelyhigh activities for C₆-aldehyde formation. However, the activitiesof the leaves decreased gradually with leaf expansion. Seedlingsand seeds containing cotyledons showed low activities for C₆-aldehydeformation, because of the occurrence of an inhibitory factorin the cotyledons. The substrate specificity of the enzymeswas essentially the same among the various developmental stagesof leaves tested. (Received February 5, 1982; Accepted March 19, 1982)     

Cytokinins in Germinating Seeds of Phaseolus vulgaris L. II. Transport and Metabolism of 8[14C]t-Zeatin Applied to the Radicle

The application of 8[¹⁴C]t-zeatin to the radicle tips of germinatingPhaseolus vulgaris seeds revealed that cytokinins are transportedrapidly from the embryonic axis to the cotyledons, and are utilizedand metabolized extensively in these organs. The informationobtained on the transportation between the different parts ofthe embryo is consistent with the view that the mobilizationof food reserves from the cotyledons is controlled by cytokininswhich originate in the embryonic axis. Tentative identificationof the radioactive metabolites formed indicate that the appliedzeatin was altered by side-chain cleavage and by substitutionto the adenine ring. Phaseolus vulgaris, bean, germination, cytokinins, transport, radicle     

The Influence of Temperature on Seed Germination in Cultivars of Common Bean

Common bean (Phaseolus vulgaris L.) is grown over a wide rangeof environments, including sites with low or high soil temperaturesat sowing time. To describe the temperature responseof seedgermination, 20 bean genotypes were evaluated using a rolledpaper towel system with 11 constant temperatures ranging from12 to 34 °C. Germination response was characterized by fittingcumulative counts using a maximum-likelihood analysis. Rateof germination increased from abase temperature (T_b) typicallynear 8 °C to an optimal development temperature (T_o) of29 to 34 °C. T_b did not differ among common bean genotypes.Mesoamerican germplasm showed slightlyhigher T_o than Andeangermplasm, but there was large variation in T_o within each ofthe two gene pools. The single accession of tepary bean (P.acutifolius) evaluated appeared to be the mosttolerant to highgermination temperatures. Key words: Common bean, seed germination rate, temperature     

The Messenger RNA Population in the Embryonic Axes of Phaseolus vulgaris During Development and Following Germination

Misra, S. and Bewley, J. D. 1985. The messenger RNA populationin the embryonic axes of Phaseolus vulgaris during developmentand following germination.—J. exp. Bot. 36: 1644–1652. Messenger RNAs were extracted from young, mid-maturation, late-maturation,mature-dry, and 20-h-germinated embryonic axes of Phaseolusvulgaris cv. Taylor's Horticultural. They were translated invitro in a rabbit reticulocyte lysate protein synthesizing system.Analysis of the translation products using two-dimensional polyacrylamidegel electrophoresis indicated that there were substantial changesin the messenger RNA populations of developing and germinatingaxes. The number of polypeptides synthesized increased sharplyat 20–22 d after pollination and then declined. Therewas a parallel increase and decrease in the Poly(A)⁺ contentof the seed axis. The analysis showed that certain messageswere present throughout development and were stored in maturedry seed. These messages were degraded upon subsequent rehydration.Some messages appeared during mid-maturation but declined duringlater stages of development and were absent from the matureseed. In the germinating seed a set of messages unique to germinationappeared. Key words: —Seed development, germination, mRNA, in vitro translation, Phaseolus vulgaris