Phenotypic Mutations Induced During Storage in Barley and Pea Seeds

Previous work on chlorophyll-deficiency mutations in pea andbarley has shown that a significant increase in mutations isinduced by storing seeds under various conditions which leadto losses of viability to about 50 per cent. The work here showsthat a detectable increase in mutation frequency is also associatedwith much smaller losses of viability. Pea seeds were storedat 35 °C and 16.5 per cent moisture content for 40 and 57d when viability fell from 99 to 93 and 82 per cent, respectively.At the same time mutation frequency (percentage of seeds containingrecessive point mutations) increased from 1.62 per cent in thecontrol treatment to about 3 to 4 per cent. Barley seeds at15.5 per cent moisture content were stored at 50 °C for42 and 54 h, and at 35 °C for 28 and 39 d. During theseageing treatments viability fell from 98 to 75, 26, 93 and 48per cent respectively and the mutation frequency increased fromzero to between about 0.3 to 0.9 per cent. In both species theinduction of mutation by ageing treatments was significant butthe differences between the various ageing treatments were not.It is concluded that there is probably no safe threshold lossof viability which completely avoids mutation, and these resultssupport the view that for genetic conservation seeds shouldbe stored under conditions which minimise loss of viability. Pisum sativum L., pea, Hordeum distichum L., barley, mutation frequency, seed storage, seed viability     

The Development of Desiccation-tolerance and Maximum Seed Quality During Seed Maturation in Six Grain Legumes

‘Physiological maturity’, i.e. the time when seedsreach their maximum dry weight during development, occurredwhen maturation drying on the parent plant in the field hadreduced seed moisture content to approximately 60 per cent infaba bean (Vicia faba L.), lentil (Lens culinaris Medic.), chickpea(Cicer arietinum L.), white lupin (Lupinus albus L.), soya bean(Glycine max [L.] Merr.) and pea (Pisum sativum L.) The onsetof desiccation-tolerance, i.e. the ability of seeds to germinatefollowing harvest and rapid artificial drying, coincided withphysiological maturity, except in pea where it occurred a littleearlier at about 70 per cent moisture content. Maximum seedquality as determined by maximum viability, minimum seedlingabnormalities and maximum seedling size occurred in pea, chickpeaand lupin when seeds were harvested for rapid drying at physiologicalmaturity; but for maximum seed quality in the other speciesmaturation drying had to proceed further - to about 45 per centmoisture content in soya bean and to about 30 per cent moisturecontent in lentil and faba bean seed crops. Much of this variationamongst the six species, however, was due to differences inthe variation in maturity within each seed crop. Results forindividual pods showed that peak maturity, i.e. maximum seedquality following harvest and rapid artificial drying, was achievedin all six species once maturation drying had reduced the moisturecontent of the seeds to 45–50 per cent. In pea, faba beanand soya bean there was a substantial decline in viability andan increase in seedling abnormalities when harvest was delayedbeyond the optimal moisture content for harvest.     

Loss of Viability in Lettuce Seeds and the Accumulation of Chromosome Damage under Different Storage Conditions

Loss of seed viability in lettuce (Lactuca sativa L.) duringstorage is associated with an increase in the frequency of cellsin the surviving seeds showing chromosome damage during firstmitoses. The relation is linear when probit of the frequencyof aberrant cells is plotted as a function of probit percentagenormal germination. The slope of the relation, however, variesaccording to moisture content so that the proportion of aberrantcells for any given loss of germination increases with decreasein moisture content over the range 13.0–5.5 per cent.At 3.3 per cent moisture content, however, the proportion ofaberrations was no greater than at 5.5 per cent moisture content;and at 18.1 per cent moisture content the proportion was noless than at 13.0 per cent moisture content. Despite these differences,the increase in chromosomal aberrations per unit time for agiven temperature was always less the lower the moisture content.Diplontic selection markedly reduced the frequency of chromosomalaberrations and eliminated the differences in these frequenciesbetween the different storage treatments. But even after fiveweeks' growth, root tips from aged seed still contained abouttwice as many aberrant cells as compared with similar root tipsderived from the original seed stock. Studies on the frequencyof recessive mutations indicated that excessive amounts of heritablemutations were not present in the progenies of aged seed, evenwhen stored at moisture contents as low as 5.5 per cent. Allthis and other evidence reinforces the view that orthodox seedsfor genetic conservation should be stored at not more than about5 per cent moisture content, and that even lower moisture contentsare worth considering. The results also emphasise the need formaintaining a high regeneration standard, i.e. the percentageto which seed viability is allowed to fall during storage beforethe seed stock is regenerated. Lactuca sativa, lettuce, seed storage, seed viability, chromosomal aberrations, phenotypic mutations     

Wheat Viability at High Moisture Content under Hermetic and Aerobic Storage Conditions

Wheat seeds (Triticum durum) were stored under both hermeticand aerobic conditions at 25 °C with moisture contents from15 to 33 per cent. Under hermetic storage, seeds lost viabilitymore rapidly the higher the moisture content, whereas in aerobicstorage, seed longevity was enhanced as the moisture contentwas increased from 24 to 31 per cent, and over this range ofmoisture content the seeds survived longer under aerobic thanhermetic storage. On the contrary, an apparent reversal of thistrend occurred when moisture content was increased above 31per cent. The possibility that the changes in longevity occurring at highermoisture contents might be due to the activation of seed metabolismwas supported by the enhanced incorporation of [³H]leucine intoTCA insoluble material (indicating increased protein synthesis)and the reduced leakage of glucose (indicating increased membranerestitution) when seeds were allowed to achieve higher moisturecontents during the prehydration period. The highest level ofseed activation was found in seeds preconditioned to about 31per cent moisture content. Moreover, these seeds, when subsequentlystored under aerobic conditions, maintained a higher rate ofprotein synthesis and lower membrane permeability during thestorage period than seeds at lower water contents. It is suggested that seeds stored at a sufficient hydrationlevel in the presence of oxygen can sustain an effective metabolismduring extended storage, thus permitting the repair of cellulardamage. However, it might be possible that at about 33 per centmoisture content seed could suffer from an excessive advancementof metabolism. Triticum durum, seed storage, effects of high moisture content and oxygen     

The Effect of Desiccation on the Longevity of Seeds of Araucaria hunsteinii and A. cunninghamii

Seeds of Araucaria hunsteinii K. Schum. dried quicker at 29°Cthan at 19°C and quicker with the seed-coat removed thanwhen intact; seeds enclosed in polyethylene bags increased inmoisture content. At 15°C, seeds in a flow of air driedquicker than seeds in a box with silica gel, which in turn driedquicker than seeds in a box with no desiccant. No loss of germinationability occurred on drying fresh seed from 53 to about 32 percent moisture content (fresh weight basis); during further desiccationthe percentage germination was related to percentage moisturecontent in the form of a sigmoid curve, culminating in a completefailure to germinate at approximately 14 per cent moisture content.A consistent relationship was observed for all treatments andthe mean critical moisture content for seed death (failure togerminate) was near 23 per cent. Excised embryos grew on 1 percent agar but died if previously subjected to 14 h of desiccationat 15°C. In contrast, no relationship was found between germination andmoisture content of A. cunninghamii D. Don on desiccation from21 to 7 per cent moisture content. Possible causes for the observeddifference in response to desiccation are discussed and methodsfor seed storage are considered. Araucaria hunsteinii, Klinkii pine, Araucaria cunninghamii, Hoop pine, desiccation, seed longevity, storage of seeds     

Water as a Storage Medium for Elaeis guineensis (pisifera) Seeds under Aseptic Conditions

Over 50 per cent germination has been obtained from Elaeis guineensisform pisifera seeds stored in unaerated sterile distilled waterfor 6 months. The moisture levels of the seeds and excised embryoswere of the same order (20–30 and 60–70 per cent,respectively) as those of fully imbibed fresh seeds. The implicationsof an apparent lower oxygen requirement by seeds stored underwater as against germinating seeds are discussed in the contextof the successful storage. Elaeis guineensis, pisifera, germination, seed dormancy, embryo     

The Influence of Genes ar and n on Senescence in Pisum sativum L.

The effect of genes ar (violet flowers, small hilum) and n (thick,fleshy pod wall) on whole plant in senescence peas was examinedby comparing Ar- with arar and N- with nn plants in segregatingprogenies. Homozygosity for ar or n significantly delayed the time whenthe plants were ready for harvest of their entire seed crop.These genes did not delay either the onset of reproduction orthe onset of apical arrest in the first instance. However, whereasAr- N- plants almost invariably senesced and died as the firstseed crop matured, the majority of arar and/or nn plants entereda period of secondary growth and a further fruiting cycle. Comparedwith Ar- plants, arar plants had over twice as many pods andseeds but individual seeds were 58 per cent lighter and totalseed yield (wt) was 19 per cent less. Pod length was unaffected.Compared with N-plants, nn plants had shorter pods (16 per cent),fewer seeds per pod (21 per cent), smaller seeds (20 per cent)and a lower total seed yield (wt 14 per cent less). It appearsthat ar and n impose a lower metabolic drain per reproductivenode as a consequence of their effects on hilum anatomy andpod morphology, respectively. These mutants disrupt the normalpattern of monocarpic senescence by breaking the coordinationbetween apical arrest and subsequent events. The developingseed crop delimited by the first arrest fails to cause plantdeath, possibly because sink size is less than in normal counterparts. Pisum sativum L, garden pea, senescence, hilum, pod, seed size, genetics     

Seed Viability Constants for Lettuce

Seeds of two lettuce cultivars (Lactuca sativa L., cv. Meikoninginand cv. Grand Rapids) were hermetically stored with constantmoisture contents ranging between 3.6 and 17.9 per cent (freshweight basis) at constant temperatures ranging between 5 and75 °C. The decline with time in percentage germination andpercentage normal seedlings was determined for each storagetreatment. The data were fitted to an equation which containsthe constants: K₁, the probit of the initial percentage germinationor normal seedlings; K_E, a species constant; C_W, the constantof a logarithmic moisture term; C_H, the constant of a lineartemperature term and C_Q, the constant of a quadratic temperatureterm. Regression analysis of data from storage periods up to5.5 years at temperatures of 5–75 °C and seed moisturecontents of 3.6–13.6 per cent yielded the following values:K_E= 8.218, C_W=4.797±0.163, C_H=0.0489±0.0050 andC_Q=0.000365±0.000056. Although this equation consistentlyprovided a better fit, simplified equations, assuming eithera log-linear relationship between seed longevity and temperature,or a log-linear relationship between seed longevity and bothmoisture content and temperature, accounted for more than 94per cent of the variation at the restricted temperature rangeof 5–40 °C. Longevity of the same seed lots at sub-zero temperatures (–5,–10 and –20 °C) was studied in separate tests.Freezing damage, resulting in abnormal seedlings in the germinationtest, occurred at –20 °C when the moisture contentof the seeds exceeded 12 per cent. No decline in percentagenormal seedlings was observed after a storage period of 18 monthsor longer at –20 °C, provided the seed moisture contentdid not exceed 9.5 per cent. For seeds stored at –5 and–10 °C with 9.6–12.5 per cent moisture content,the observed rate of decline of percentage normal seedlingswas adequately predicted by the viability equation, using theabove values for the constants. This suggests that for low moisturecontents the viability equation can be applied to estimate longevityat sub-zero temperatures. Lettuce, Lactuca sativa (L.), seed longevity, seed storage, viability constants, storage conditions     

The Influence of Gaseous Environment on the Storage Life of Araucaria hunsteinii Seed

The effect of various gas mixtures on the longevity of hydratedseeds of Araucaria hunsteinii K. Schum. was assessed under controlledconditions. The length of storage life decreased as oxygen concentrationwas reduced from 21 to zero per cent. No effect of carbon dioxideon seed longevity was detected within the range 1–50 percent when combined with 10 or 21 per cent oxygen. Ethylene at0.01 per cent, and sealed foil or polyethylene bag storage reducedthe period of seed germinability compared with that for 21 percent oxygen. Ethanol accumulation took place in stored seedswhen the environmental oxygen concentration was below a thresholdvalue which lay between 1 and 5 per cent. It is proposed that the observed effects of gases on longevityof hydrated seeds may be mediated through an influence on aerobicrespiration rate. Practical implications of the results areconsidered. Araucaria hunsteinii, Klinkii pine, seed longevity, seed storage, gas environments, oxygen, carbon dioxide, ethanol accumulation     

Nitrogen Fixation in the Phyllosphere of Tropical Plants: Occurrence of Phyllosphere Nitrogen-Fixing Micro-organisms in Eastern India and their Utility for the Growth and Nitrogen Nutrition of Host Plants

Of the 560 leaf samples belonging to 259 species of green plantsexamined more than 50 per cent of the Angiosperms and 25 percent of the Pteridophytes and Gymnosperms revealed the presenceof N₂-fixing micro-organisms in their phyllosphere. Plants particularlyremarkable in this respect are orchids and several other epiphytes,Scindapsus officinalis, Ficus and cucurbits. Most of the isolatesappear to be biotypes of Klebsiella pneumoniae. The more activestrains fixed more than 5 mg N g^–1 glucose utilized andreduced more than 100 nmol C₂H₂ mg^–1 cell d. w h^–1. The efficacy of the phyllosphere N₂-fixing isolates for N-nutritionof host plants was studied by spraying suspensions of the culturesgrown on N-free media on rice and wheat seedlings. In IR-26rice or Sonalika and Janak wheat grown on soil in wooden flatsor earthenware pots, 22 per cent of the 161 cultures studiedcaused increased height and about three-quarters of the culturesenhanced dry weight by more than 50 per cent; chlorophyll andN-contents were enhanced more than 50 per cent by about halfand two-thirds of the cultures respectively. In N-free sandculture 26 of the 50 promising strains doubled N-content, and30 doubled dry weight of the tested plants. In some cases dryweight, number of grains per panicle, and 1000 grain weightwere increased by 300, 70–83 and 126–158 per centrespectively; N-content of straw and seed was increased three-or fourfold. In several cases the beneficial effects were foundto match closely the performance of plants receiving ammoniumsulphate. Nitrogen-fixing micro-organisms, nitrogen nutrition, phyllosphere, rice, tropical plants, wheat     

The Function of the Hilum in Some Papilionaceae in Relation to the Ripening of the Seed and the Permeability of the Testa

In seeds of Trifolium repens, T. pratense, and Lupinus arboreus,the hilum is a hygroscopically activated valve in the impermeableepidermis of the testa. When relative humidity was low the fissurein the hilum opened permitting the seed to dry out; when therelative humidity was high the fissure closed obstructing theabsorption of moisture. During seed-ripening the moisture contentfell readily to approximately 25 per cent., and thereafter moreslowly until the epidermis became impermeable at approximately14 per cent, moisture content. Further drying of the seed tookplace only by diffusion of water vapour through the hilum. ‘Hard’seeds tended to have a moisture content in equilibrium withthe lowest relative humidity to which they had been exposed.They absorbed moisture under conditions of gradually increasingrelative humidity such that the hilar fissure remained open.The duration of the impermeable condition increased with thedegree of desiccation brought about by loss of water throughthe hilum.     

The Effect of Temperature and Moisture Content on the Longevity of Seed of Ulmus carpinifolia and Terminalia brassii

Seeds of the Smooth-leafed Elm (Uimus carpinifolia) and of thetropical forest tree Terb (Terminalia brassii) were stored hermeticallyand sampled at intervals for periods of up to two years. Bothspecies possess ‘orthodox’ seed (increasing longevityis observed as either moisture content or temperature are reduced)within the temperature ranges from — 13 to 52°C (Elm)and from —4 to 42°C (Terb) and within the moisturecontent ranges from 3 to 19 per cent (Elm) and from 5 to 14per cent (Terb) on a fresh weight basis. Elm seed stored at—75°C showed the expected relationship between longevityand moisture content, but did not differ significantly in longevityfrom seed kept at — 13°C when moisture contents wereheld constant. Probit analysis of the relationship between germinationpercentage and time was performed for each storage environment,yielding a slope from which the standard deviation of the distributionof seed deaths over time () was calculated. Standard deviationvalues were used in turn to determine the values of constantsin a viability equation which had previously been applied toseed of barley, chickpea, cowpea and soybean. The equation,which gave a good fit to the results obtained, can be used topredict viability for seed in storage over a wide range of environmentalconditions. Some limitations to the applicability of the viability equationwere defined. At 22 per cent and higher moisture contents Elmseed survived longer than predicted. Furthermore, all Elm andTerb seed was killed quickly on placing in —75°C at22 and 20 per cent moisture content respectively, but high viabilitywas retained for several days at 19 and 17 per cent respectively.Practical implications of the results are discussed. Uimus carpinifolia Gleditsch, Smooth-leafed Elm, Terminalia brassii Exell, Terb, seed longevity, seed storage, moisture content, temperature     

Microbiological Synthesis of Fat: THE EFFECTS OF VARYING PH, TEMPERATURE, NITROGEN, INCUBATION PERIOD, AND SUGAR CONCENTRATION ON FAT-PRODUCING MOULDS

1. A more detailed study has been made of the influence of thesefactors on fat formation by Aspergillus nidulans, Penicilliumspinulosum, and Penicillium javanicum. 2. The effect of halving the glucose, while keeping the ammoniumnitrate concentration constant, lowered the yield of fat onsugar used in A. nidulans and P. spinulosum but not in P. javanicumcultures. 3. Keeping the same N: C ratio and raising the glucose concentrationfrom ro to 20 per cent. showed that to per cent. glucose wasmore efficiently converted to fat by A. nidulans and P. javanicum. 4. The iodine values of the extracted fats were higher, in general,with increased length of incubation. Low ammonium nitrate concentrations,however, tended to give low iodine values. 5. The results have been applied on a larger scale by growthin Roux bottles, Glaxo flasks, and a flat stainless-steel tank.     

Logarithmic Relationship between Moisture Content and Longevity in Sesame Seeds

The relationship between seed moisture content and seed longevityin sesame (Sesamum indicum L.) in hermetic storage at 50 °Cis logarithmic. The logarithmic relationship is maintained from15 per cent down to 2 per cent – the lowest moisture contenttested — but above 15 per cent this ‘air-dry’relationship no longer holds since further increase in seedmoisture content does not reduce longevity. Tentative estimatesof constant values for the improved seed viability equationare provided, and implications for long-term storage are discussed. Sesame, Sesamum indicum L., seed storage, improved viability equation, seed moisture content, seed longevity prediction     

Microbiological Synthesis of Fat: THE FORMATION OF FAT FROM SUCROSE

1. Forty-three strains of moulds, selected from ten species,were examined to ascertain their capabilities as producers offat, when grown in four different solutions of nutrient inorganicsalts together with sucrose. The three highest yields of feltwere given by Penicillium javanicum van Beyma, P. Soppi Zaleski,and Aspergillus nidulans Eidam, in that order. The fat contenton felt weight was maximal at 34·8 per cent, with P.Soppi, at 28·4 per cent. with Fusarium lini(1), and at25·8 per cent. with A. nidulans; on sugar utilized itwas maximal at 11·4 per cent. with P. Soppi, at 7·9per cent, with A. nidulans, and at 5·6 per cent. withF. lini(1). 2. Other moulds considered worthy of further trial were P. spinulosum,P. aurantiobrunneum, F. graminearum, A. flavus, F. oxysporum,A. clavatus, and P. oxalicum.     

Succinoxidase and Cytochrome Oxidase in Mitochondria from the Spadix of Arum

1. Mitochondria prepared from young Arum spadix have enoughcytochrome c oxidase to account for the rate at which succinateis oxidized, but succinoxidase activity increases markedly asthe plants mature so that in old material cytochrome oxidaseactivity is only 10 per cent, of succinoxidase. 2. Disintegration of the mitochondria by vibration with ballotini,treatment with digitonin or incubation in the warm reveals anintra-mitochondrial cytochrome c oxidase probably active enoughto account for the fastest rates of succinate oxidation. 3. Succinic dehydrogenase activity is demonstrated and experimentswith p-chloromercuribenzoate indicate that it plays a part inthe oxidation of succinate. 4. Cyanide completely inhibits both external and internal cytochromeoxidase but even at the earliest stages it only reduces succinoxidaseby about 50 per cent. Antimycin A also inhibits succinoxidaseby about 50 per cent.     

Minerals, Carbohydrates and Amino Acids of Pollens from some Woody and Herbaceous Plants

The mineral, carbohydrate, protein, amino acid, lipid and energycontents of the seven main pollens gathered by honey-bees insouth-east Scotland were determined. Neither dry matter (mean73 per cent) nor ash (mean 3 per cent) were very variable. Thecalcium content of all pollens was relatively similar as werethe magnesium, potassium and sodium contents but manganese whichwas relatively high in ericaceae and phosphorus which was lowin Trifolium were more variable. Acer pollen contained mostprotein and Fagus least, and of the amino acids only serine,cystine and histidine were significantly different in the differentpollens. Over 30 per cent of all pollens was a mixture of fructoseand glucose, ericaceae containing most and Acer least. Hemicellulose(mean 7 per cent) was high in Ranunculus and low in Acer, cruciferaeand Fagus. ‘Cellulose’ (mean 0•5 per cent)was high in cruciferae and low in ericacea. Ether extract (mean5 per cent) was high in cruciferae and Fagus. The mean grossenergy of pollen was 23 kJ g^–1. Fresh pollen is comparable with lean meat as a source of energyand protein, and is more like dormant plant seeds than herbagein its water, ash and protein content but stores energy as sugarrather than starch or lipid.     

CHANGES IN THE CONTENT OF GIBBERELLIN-LIKE SUBSTANCES IN RIPENING SEED AND POD OF LUPINUS LUTEUS

The amount of gibberellin-like substances in seed, pod, embryoand "endosperm" of Lupinus luteus in relation to their developmentwas studied by means of the rice seedling test. The amount of gibberellin-like substances per seed increasedremarkably at the first stage of development, when the growthof seed and pod was very slow, and attained its maximum at suchan early stage as the dry weight of seed reached a value ofonly 10 per cent and that of pod 28 per cent of their respectivemaximum levels. Changes in amount of gibberellin-like substancespresent in the pod were very closely related to the changesin growth of pod. Gibberellin-like substances equivalent to0.3 µg gibberellin A₃ per seed and 0.028 µg perpod were found at the 20th day after anthesis, and no activityin mature seed and 0.09 µg per pod at the 50th day. Noactivity was found in the embryo itself at all stages, indicatingthat the gibberellin-like substances in seed occur only in the"endosperm," i.e. in the tissues that will eventually form testain fully ripened stage. (Received December 18, 1962; )     

Improved Equations for the Prediction of Seed Longevity

Equations for predicting seed longevity in storage have beenimproved so that they now take into account variations withina species in initial seed quality—which is affected bygenotype and pre-storage environment—and so that theyare more accurate over a wider range of storage environmentsThese improvements have been incorporated into a seed viabilitynomograph for barley (Hordeum distichum L.) which may be usedto predict percentage viabihty of any seed lot after any timein any storage environment within the range –20 to 90°C and 5–25 per cent moisture content. Applicationsof the improved equations to seed drying and to long-term seedstorage for genetic conservation are discussed. Hordeum distichum L., barley, seed viability, seed longevity prediction, seed storage, seed drying, storage temperature, seed moisture content, genetic resources conservation     

Microbiological Synthesis of Fat: PRELIMINARY SURVEY OF THE FAT-PRODUCING MOULDS

1. A survey of potential fat-producing moulds has shown that atleast 40 strains from 10 species are of interest.
2. These havebeen grown on five different media and A. nidulans,P. spinu-losum,P. javanicum, P. piscarium, P. flavo-cinereum,P. oxalicum,A. flavus, A. flavipes have shown the most promise.
3. The fatcontent on felt weight was maximal at 39?7 per cent,with A.flavipes and 34?6 per cent, with F. lini, and on usedsugarwas maximal at 9?3 per cent, with A. flavipes and 6?7per cent,with A. nidulans.
4. The potential value of micro-organisms asfat producers is discussedwith reference to the moulds.